Bioresource Technology 240 (2017) 25–32

Contents lists available at ScienceDirect

Bioresource Technology
journal homepage: www.elsevier.com/locate/biortech

Alkali-assisted membrane cleaning for fouling control of anaerobic

ceramic membrane bioreactor
Xiaojie Mei a,b, Pei Jun Quek b, Zhiwei Wang a, How Yong Ng b,⇑
a
State Key Laboratory of Pollution Control and Resource Reuse, School of Environmental Science and Engineering, Tongji University, 1239 Siping Road, Shanghai 200092, PR China
b
Centre for Water Research, Department of Civil and Environmental Engineering, National University of Singapore, 7 Engineering Drive 1, Singapore 117576, Singapore

h i g h l i g h t s

 An alkali-assisted CEB cleaning method in AnMBR was developed.

 0.05–1.30 mmol-NaOH/L had positive impacts on anaerobic biomass under short-term exposure.
 10–20 mmol-NaOH/L backflushing significantly reduced membrane fouling rates and slightly improved methanogenic activities.
 12 mmol-NaOH/L is cost-efficient and fouling-control dosage.

a r t i c l e i n f o a b s t r a c t

Article history: In this study, a chemically enhanced backflush (CEB) cleaning method using NaOH solution was proposed
Received 27 December 2016 for fouling mitigation in anaerobic membrane bioreactors (AnMBRs). Ex-situ cleaning tests revealed that
Received in revised form 11 February 2017 NaOH dosages ranging from 0.05 to 1.30 mmol/L had positive impacts on anaerobic biomass, while
Accepted 14 February 2017
higher dosages (>1.30 mmol/L) showed inhibition and/or toxic impacts. In-situ cleaning tests showed
Available online 16 February 2017
that anaerobic biomass could tolerate much higher NaOH concentrations due to the alkali consumption
by anaerobic process and/or the buffering role of mixed liquor. More importantly, 10–20 mmol-NaOH/L
Keywords:
could significantly reduce membrane fouling rates (4–5.5 times over the AnMBR with deionized water
Sodium hydroxide
Membrane fouling
backflush) and slightly improve methanogenic activities. COD removal efficiencies were over 87% and
Ceramic membrane peaked at 20 mmol-NaOH/L. However, extremely high NaOH concentration had adverse effects on
Anaerobic membrane bioreactor filtration and treatment performance. Economic analysis indicated that 12 mmol/L of NaOH was the
Domestic wastewater cost-efficient and optimal fouling-control dosage for the CEB cleaning.
Ó 2017 Elsevier Ltd. All rights reserved.

1. Introduction matters (DOMs) on the membrane surface (Le-Clech et al., 2006),

Anaerobic membrane bioreactor (AnMBR) has high potential to
be a good technology that can achieve energy neutrality in
wastewater treatment in the near future. (McCarty et al., 2011;
Scherson and Criddle, 2014; Smith et al., 2014). In addition to the
energy that can be recovered from methane-rich biogas, the advan-
tages of anaerobic treatments over conventional aerobic processes
include lower energy consumption and reduced sludge production
(Ozgun et al., 2013; Stuckey, 2012). Coupling ceramic membranes
into an anaerobic bioreactor further enhances its practical applica-
tion as ceramic membranes are resistant to corrosive chemicals
and harsh environmental conditions (Arslankaya, 2016; Lee et al.,
2013b). However, membrane fouling, which is caused by the
deposition of sludge flocs and the adsorption of dissolved organic
⇑ Corresponding author.
E-mail address: howyongng@nus.edu.sg (H.Y. Ng).
is regarded as one of the major barriers hindering the application
of AnMBR to domestic wastewater treatment (Choo and Lee,
1996; Yue et al., 2015). Chemical cleaning is an essential approach
to address fouling issues and several protocols including in-situ
maintenance cleaning, in-situ recovery cleaning and ex-situ clean-
ing are generally applied in aerobic MBRs (AeMBRs) (Wang et al.,
2014a; Wei et al., 2011). Although some of the cleaning protocols
have been successfully implemented in lab-scale or scaled-up
AeMBRs (Wang et al., 2014b; Wei et al., 2011; Zhou et al., 2014),
the cleaning mechanism and application in AnMBRs are still
lacking.
Among the prevalent cleaning protocols, chemically enhanced
backflush (CEB), a maintenance cleaning that combines conven-
tional physical backflushing with chemical cleaning in a single pro-
cess, is of great importance for operation of AnMBRs. Typically, CEB
is performed over long operation intervals (i.e., weekly or monthly)
and with high chemical loadings (e.g. 500–3000 mg/L NaClO)

http://dx.doi.org/10.1016/j.biortech.2017.02.052
0960-8524/Ó 2017 Elsevier Ltd. All rights reserved.
26 X. Mei et al. / Bioresource Technology 240 (2017) 25–32
(Ramos et al., 2014; Walczak and Cywinska, 2007; Wei et al.,
2011). The chemical reagent in the backflushing solution would
diffuse and/or permeate through the membrane into the mixed
liquor. Therefore, the microbial activities and biological perfor-
mance might be affected by the exposure to chemical reagents
(Lee et al., 2013a).
Sodium hypochlorite (NaClO) is a widely-used cleaning reagent
applied in in-situ and ex-situ cleaning due to its chemical stability
and good cleaning efficiency (Judd, 2010; Lee et al., 2016; Ramos
et al., 2014; Wei et al., 2011). However, NaClO, being a kind of bio-
cide, may significantly affect the performance of the bioreactors
due to the inhibition of bacterial activities (Walczak and
Cywinska, 2007). According to Han et al. (2016), NaClO concentra-
tions exceeding 1 mg/g-SS would damage microbial cells and inhi-
bit enzyme activities. Sodium hydroxide (NaOH) is also widely
used as a cleaning reagent in chemical cleaning (Ang et al., 2011;
Tian et al., 2010; Zhou et al., 2014). It has been reported that NaOH
is effective in removing protein-related foulants by disintegrating
large organic particles (such as colloids) into fine particles and
hydrolyzing organic matters into small molecules (Wang et al.,
2014c; Yu et al., 2013a). However, to date, detailed information
on NaOH backflushing for fouling control in AnMBR is rare, the
consequence effects to the bioactivities in the reactor by NaOH
backflushing is also not clear yet.
It is well-known that alkali can hydrolyze organic matters (e.g.,
proteins and carbohydrates) into small molecules (e.g., amides and
monosaccharides) (Yu et al., 2013a), while a high background alka-
linity could disintegrate the anaerobic biomass and damage the
functional proteins of enzymes (Li et al., 2008; Wang et al.,
2014a). Dehydrogenase and coenzyme F420 are two key enzymes
involved in the acetogenic and methanogenic process. Specific
methanogenic activity (SMA) and biochemical methane potential
TMP meter Suction pump
Feed pump
Control box
Stirrer
Level
sensor
Ceramic
membrane
Sampling
port
Gas
diffuser
Feed tank AnCMBR
Fig. 1. Schematic diagram of the AnCMBR setup.
(BMP) are closely related to methane production, which are widely
used to evaluate the methanogenic performance. However, few
studies have carried out to investigate the bioactivity under rele-
vant NaOH dosages for membrane cleaning.
The objectives of this present work are therefore to elucidate
the role of NaOH backflushing (CEB) in membrane fouling mitiga-
tion and its impact on reactor performance. Ex-situ and in-situ
cleaning tests were carried out. Cleaning mechanism and microbial
responses to chemical cleaning were discussed based on the anal-
ysis of organic components and the detection of bioactivity
indicators.
2. Materials and methods
2.1. Reactors setup
Two identical AnCMBRs (named as R1 and R2), each with an
effective volume of 3.6 L, were operated in parallel (Fig. 1). A flat-
sheet ceramic membrane module (ItN Nanovation AG, Germany)
with a mean pore size of 80 nm and effective area of 0.08 m2 were
immersed in each AnCMBR. Three peristaltic pumps (BT100,
Longer, China) were used to feed influent into the bioreactor,
extract the permeate at a flux of 8 L/(m2
h) and pump backflushing
solution to clean the membrane at a flux of 20 L/(m2
h). A dia-
phragm gas pump (N022 STE, KNF, Germany) was used to recircu-
late biogas for membrane surface scouring. The operation mode
was set as 10 min for suction and 1 min for backflushing.
2.2. Reactor operation conditions
The AnCMBRs were operated under the ambient temperature of
25–30 °C. The operating hydraulic retention time (HRT) and solids
Permeate
Backwashing pump
Backwashing
solution
Biogas
Water line
Gas line
PLC control line
Gas pump Gas collector
 X. Mei et al. / Bioresource Technology 240 (2017) 25–32
retention time (SRT) were maintained at 5.8 h and 60 days, respec-
tively, which were the optimal operation conditions according to
our previous study (Yue et al., 2015; Mei et al., 2016). The
AnCMBRs were fed with domestic wastewater from a wastewater
reclamation plant in Singapore. The feedwater had a total COD of
417 ± 61 mg/L, soluble COD of 92 ± 28 mg/L, TN of 56.4 ± 5.6 mg/
L, NH+4-N of 38.7 ± 4.5 and suspended solids (SS) of 239 ± 61 mg/L.
During the startup period of 30 days, ex-situ cleaning were carried
out every 15 days using NaOH at 10 mmol/L concentration when
the TMPs were about 40 kPa. Subsequently, in-situ cleaning tests
using different NaOH dosages were carried out in the 6 phases of
this study.
2.3. Ex-situ cleaning test
To evaluate the cleaning effect of NaOH on foulants detachment
from the membrane surfaces, the fouled membrane was collected
from Phase 1 when the TMP was >40 kPa, flushed with tap water
to remove the cake layer and soaked in a separate cleaning tank
(Fig. S1 of Supporting Information). To figure out the cleaning
mechanism, the fouled membrane was soaked in alkaline solution
(10 mmol/L NaOH) followed by acid solution (10 mmol/L HCl), and
the soaking duration was 1 day for each chemical solution. The
total organic carbon (TOC), proteins (PRO), polysaccharides (PS),
humic acids (HA) and inorganics (such as magnesium (Mg) and cal-
cium (Ca)) in the cleaning solution were measured. Fractions of
dissolved organic matters (DOMs) were further determined by liq-
uid chromatography-organic carbon detection (LC-OCD, Dr. Huber,
DOC-LAB) to elucidate the cleaning mechanism. Prior to the analy-
ses, the pH values of all samples were adjusted to approximately 7.
To evaluate the biological effect of NaOH on biomass activities
under short-term exposure, biomass activity indicators including
SMA, BMP and activities of dehydrogenase (DHA) and coenzyme
F420, were tested under a series of NaOH dosages ranging from
0.001 to 20 mmol/L.
2.4. In-situ cleaning test
To investigate the cleaning effect of NaOH on the fouling control
in the long-term operation of AnMBR, a series of NaOH solutions
(i.e., 1, 5, 10, 20, 50 mmol/L) were tested to determine the opti-
mized cleaning concentration. Control test with deionized (DI)
water as the backflushing solution was also carried out. The oper-
ating parameters of the six phases are listed in Table S1.
2.5. Biomass activity indicators
SMA and BMP were measured according to a previous publica-
tion (Mei et al., 2016). The biomass samples were taken from the
reactor during Phase 1. Prior to the test, biomass samples were
washed with DI water 3 times, centrifuged at 1000g for 10 min
to remove external substrate and resuspended in DI water. Nutri-
ents were added according to a previous publication (Jeison,
2007). NaOH was added to nutrient solution to form a series of
concentrations and each test was done in duplicate. Throughout
Table 1
Foulants distribution in membrane pore blocking.
Unit: mg/m2 PRO
Run 1 NaOHremoved 109.6
HClremoved 8.0
Run 2 NaOHremoved 116.8
HClremoved 10.2
Run 3 NaOHremoved 182.5
HClremoved 5.1
27
the test duration, the serum bottles were incubated at 35 °C and
agitated at 125 rpm. Methane content in the head space was deter-
mined by gas chromatography (GC-2010PLUS, Shimadzu, Japan)
equipped with a barrier ionization discharge (BID) detector. The
SMA (mL/(gVSS
d)) was calculated by dividing the volume of
methane produced per unit time (maximum slope of cumulative
methane with time) by the initial weight of VSS (g) in the SMA bot-
tles. The BMP (mL/gVSS) was calculated according to a modified
Gompertz three-parameter model (Mei et al., 2016).
Additionally, the activity of key enzymes, namely, dehydroge-
nase and coenzyme F420, were measured after 24-h exposure to
NaOH, according to the methods published previously (Burdock
et al., 2011; Hutschemackers et al., 1982).
2.6. Sampling and analytical methods
Effluent was collected after 5 min of suction when the effluent
pH is stable (the variation of effluent pH in one suction cycle is doc-
umented in Fig. S2) and biomass was filtrated with 0.45-lm filter
papers. COD, MLSS and MLVSS were tested according to the Stan-
dard Methods (APHA, 2012). Total organic carbon (TOC) and total
nitrogen (TN) were measured by a total organic carbon analyzer
(TOC-VCSH, Shimadzu, Japan). Polysaccharides were analyzed using
phenol-sulfuric acid method (Dubois et al., 1956) using glucose as
the reference, while proteins and humic substances were mea-
sured by corrected Lowry method (Frølund et al., 1995) using
bovine serum albumin as the reference. Inorganic matter, such as
Mg2+ and Ca2+, were measured by ion chromatography (DX-500,
Dionex, USA).
3. Results and discussion
3.1. Ex-situ cleaning tests
3.1.1. Cleaning effects of NaOH on fouled membranes
To investigate the cleaning effect of NaOH on pore-blocking fou-
lants removal, membrane resistance distribution of fouled mem-
brane (Table S2) and foulants distribution in cleaning solutions
(Table 1) were measured in several runs. Table S2 indicates that
NaOH + HCl could remove almost all pore-blocking foulants and
recover membrane resistance to the initial value. Table 1 shows
that NaOH could remove about 92% of proteins and almost all
humic acids while it was inefficient in eliminating metal deposi-
tion in membrane pores. However, roughly 85% of calcium and
95% of magnesium were removed by HCl. Besides, it is worth not-
ing that acid cleaning could further remove organic foulants (from
TOC results) because acid cleaning eliminates inorganic fouling by
breaking down the mineralization between biopolymer and salts
(Wang et al., 2014a).
To elucidate the cleaning mechanism, LC-OCD chromatography
was employed to analyze the DOMs in the cleaning solutions and
the results are presented in Table 2. It shows that the concentration
of dissolved organic carbon (DOC) in the NaOH solution was much
higher than that of the HCl solution, confirming that NaOH was
effective in DOC removal. DOC was further subdivided into
HA PS TOC TN Mg Ca
348.3 79.6 203.9 23.5 1.2 19.2
– 9.9 18.0 5.6 52.3 118.6
108.6 47.2 215.1 20.8 0.8 17.0
– 7.4 13.1 7.0 25.7 92.1
293.6 68.3 281.4 31.3 1.4 14.9
– 12.4 12.7 3.3 22.1 113.0
28 X. Mei et al. / Bioresource Technology 240 (2017) 25–32

Table 2
An example of DOM fractions in the cleaning solutions.

hydrophobic (HPO) and hydrophilic (HPI) organic carbon. Accord- the NaOH dosage to 10 mmol/L, the SMA was dramatically reduced
ing to previous publications (Liang et al., 2007; Tang et al., 2010; to 0.64 mL CH4/(gVSS
d).
Yue et al., 2015), HPI are generally considered as the main foulants The BMP showed a similar trend as the SMA, with extremely
and contributed to faster fouling rates and more irreversible foul- high NaOH dosage significantly affecting methane production.
ing. In this study, most HPI were detached from membrane pores Combined with the result of SMA, it can be concluded that NaOH
by alkali and dissolved in the cleaning solution, as shown in Table 2. dosage ranging from 0.05–1.30 mmol/L had positive effects on
Under the subcomponents of HPI, except for biopolymers, all other methanogenesis, while relatively high dosage of NaOH
peaks were affiliated with low molecular weight neutrals (LMW- (>1.30 mmol/L) had inhibited and/or toxic effect on methane
N), humic substances (HS) and building blocks (breakdown prod- production.
ucts of humics), with molecular weights (MW) smaller than Dehydrogenase and coenzyme F420 are involved in acetogenesis
1 kDa (Simon et al., 2013). It has been reported that at alkaline con- and methanogenesis respectively, playing pivotal roles in anaero-
ditions, large organic particles (such as colloids) and microbes can bic metabolism (Dolfing and Mulder, 1985; Kardos et al., 2009).
be disintegrated into fine particles, soluble organic matters and/or Variations of DHA and F420 activities in the biomass samples with
low MW substances (Yu et al., 2013b). Therefore, the efficient different NaOH dosages are shown in Fig. 2(b), indicating that both
cleaning by NaOH was achieved by breaking up large molecular DHA and F420 activities maintained stable and active at NaOH
organics into soluble small molecules, which were then released dosage below 0.05 mmol/L, and then slightly increased with
from membrane pores. increasing dosage of NaOH, peaking at the NaOH dosage of
1.30 mmol/L followed by a rapid decline at NaOH dosage above
3.1.2. Biological effects of NaOH on anaerobic biomass activities 1.60 mmol/L. The high NaOH dosage (10 mmol/L) resulted in the
In anaerobic treatment processes, SMA indicates the methane- denaturation of the enzymes. Previous studies have shown that a
producing capability of anaerobic biomass and BMP is a measure high background alkalinity could disintegrate the anaerobic bio-
of total methane production, which indicates substrate biodegrad- mass, with NaOH diffusing into the microbial cells and damaging
ability (Castro et al., 2002; Owen et al., 1979). Fig. 2(a) shows the the functional proteins of enzymes (Li et al., 2008; Lin et al.,
variation of SMAs and BMPs under a series of NaOH dosages. It 1997). In addition, coenzyme F420 activity was more sensitive to
can be seen that there was no obvious improvement of SMA at NaOH compared to DHA. The maximum F420 activity was 2.5 times
NaOH concentrations below 0.05 mmol/L (averaged at 23.5 mL of the initial activity while the maximum DHA roughly increased
CH4/(gVSS
d)). At the NaOH dosages ranging from 0.05 to by 15% compared to the control test. Meanwhile, the high concen-
1.30 mmol/L, SMA increased slightly with NaOH dosages and tration of NaOH caused more significant inhibition of F420 activity.
peaked at the concentration of 1.30 mmol/L (27.9 mL CH4/ The activity of F420 at the NaOH dosage of 10 mmol/L was com-
(gVSS
d)). However, negative impacts on SMA was observed when pletely inhibited while about 65% of activity was maintained for
the NaOH dosage was above 1.60 mmol/L. With further increase of DHA.

(a) (b)
35 700 7 350
SMA BMP DHA Coenzyme F420
30 600 6 300
Coenzyme F420 (nmol/gVSS)
SMA (mL CH4/gVSS·d))

BMP (mL CH4/gVSS)

25 500 5 250
DHA (mg/gVSS·h))

20 400 4 200

15 300 3 150

10 200 2 100

5 100 1 50

0 0 0 0
0.0005 0.005 0.05 0.5 5 50 0.0005 0.005 0.05 0.5 5 50
NaOH dosage (mmol/L) NaOH dosage (mmol/L)

Fig. 2. Bioactivities of anaerobic biomass under various NaOH dosages. (a) SMAs and BMPs, (b) DHA and coenzyme F420. (Error bars, where absent, fall within symbols).
 X. Mei et al. / Bioresource Technology 240 (2017) 25–32 29

Fig. 3. TMP profiles of six phases in two AnCMBRs.

Fig. 4. COD removal rates, organic and inorganic matters in the effluent of six phases.

3.2. In-situ cleaning tests
3.2.1. Filtration performance of AnCMBRs in different phases
Fig. 3 shows the evolution of TMP during six phases. As shown
in Fig. 3, the control test had much higher TMP increase rate
(3.28 ± 0.65 kPa/d on average) than the phases with NaOH back-
flush (0.59–2.33 kPa/d), suggesting that alkali cleaning has remark-
able impact on membrane permeability. With increasing NaOH
dosage in the backflushing solution, the fouling tendencies were
generally alleviated and the lowest fouling rate (0.59 kPa/d) was
achieved at a NaOH dosage of 20 mmol/L. However, the extremely
high NaOH concentration of 50 mmol/L had an adverse effect on
filtration performance. This may be due to biomass disintegration
and the release of extracellular and/or intercellular substances,
which will be further discussed in the following section.
3.2.2. Pollutants removal and DOMs in the effluent
During the entire operation period, the MLSS concentration in
the two AnCMBRs were kept consistent (Fig. S3). The pH in the
two reactors remained at a stable level (6.6–7.4) (Fig. S2) due to
the buffering role of the mixed liquor and the alkali supplement/-
consumption by the anaerobic digestion process. The COD removal
performance and effluent characteristics of the six phases are
depicted and detailed in Fig. 4 and Table S3. From Fig. 4, the aver-
age COD removal efficiencies reached above 87% in all phases and
were slightly improved in phases 2–5, indicating that NaOH back-
flushing played a positive role in pollutants removal in AnCMBRs.
Nevertheless, backflushing with extremely high NaOH concentra-
tion of 50 mmol/L had an adverse effect on COD removal.
In view of the importance of DOMs, protein and polysaccharide
concentrations in mixed liquor and effluent were measured. As
30 X. Mei et al. / Bioresource Technology 240 (2017) 25–32

(a) (b)
7 210 35 700
DHA coenzyme F420 SMA BMP
6 180 30 600

SMA (mLCH4/gVSS·d))
Coenzyme F420 (nmol/gVSS)

BMP (mLCH4/gVSS)
DHA (mg/gVSS·h))

5 150 25 500

4 120 20 400

3 90 15 300

2 60 10 200

1 30 5 100

0 0 0 0
0 1
DI 21 53 4
10 5
20 6
50 7 0 1DI 21 35 10
4 5
20 6
50 7
NaOH dosage (mmol/L) NaOH dosage (mmol/L)
Fig. 5. Analysis of key enzymes (a) and biomass activities (b) in different phases.

shown in Fig. 4 and Table S3, the protein contents in the effluent alkali condition of 50 mmol/L. This result did not correlate well
decreased with increasing NaOH dosages and reached the lowest with the F420 activities. Although coenzyme F420 has been detected
value at a NaOH concentration of 10 mmol/L, while the polysaccha- in all methanogenic bacteria (Dolfing and Bloemen, 1985), it can-
rides in the effluent remained at very low concentrations and had not be a indicator of the total methanogenic activity but rather a
no obvious difference in the six phases. As for inorganic removal, parameter reflecting the behaviours of hydrogenotrophic metha-
there was hardly any reduction by either membrane interception nogenic microbes, as reported previously (Dolfing and Mulder,
or microbial process (Table S3). 1985; Zabranska et al., 1985). On the other hand, acetotrophic
In light of the effectiveness of NaOH on proteins removal, the methanogenesis is widely regarded as the predominant pathway
components of DOM in the effluent was further analyzed by LC- for treating low strength wastewater (Demirel and Scherer,
OCD. Among the identified peaks shown in Fig. S4, the small peak 2008). Therefore, SMA is a comprehensive index to evaluate the
for biopolymers – consisting mainly proteins, polysaccharides and overall methanogenic performance (including acetotrophic and
organic colloids (Rosenberger et al., 2006) – indicated that these hydrogenotrophic methanogenesis), while coenzyme F420 is a typ-
could be largely intercepted by the membrane. Humic substances ical index to assess the activity of hydrogenotrophic microorgan-
and building blocks were the main components in the effluent, isms. It can be inferred from this study that hydrogenotrophic
and the integrated areas of these two fractions show a decreasing methanogens were more vulnerable than acetotrophic methano-
trend with increasing NaOH concentration (below 10 mmol/L). gens when exposed to high concentrations of NaOH solution. How-
However, when the NaOH dosage exceeded 10 mmol/L, more ever, BMP shows no difference in all phases, which might be due to
humics, building blocks and low molecular neutrals were released the good biodegradability of domestic wastewater.
into the effluent, causing adverse effect on treatment (Fig. 4) and
filtration performance (Fig. 3). It is noteworthy that at a NaOH
3.3. Economic analysis
dosage of 20 mmol/L, more low MW substances appeared in the
effluent while the fouling rate remained relatively low (Fig. 3). This
To figure out the cost-efficient NaOH dosage, economic analysis
might be resulted from the hydrolysate of membrane foulants,
was performed based on the operation conditions presented above.
which can not be intercepted by membrane.
It was assumed that the unit NaOH price for industrial use was
0.5 US dollar/kg. The chemical cost of the six phases were calcu-
3.2.3. Anaerobic biomass activities under long-term exposure to NaOH
lated and documented in Fig. 6. It can be seen that the chemical
Microbial activities associated with methanogenesis were fur-
ther determined during the long-term operation. Fig. 5(a) presents 3.5 0.25
the variations of DHA and F420 activity of anaerobic biomass under Fouling rate Chemical cost
different NaOH concentrations. It can be observed that DHA and 3.0
Chemical cost (US dollar/m3)

F420 activity remained at stable and active levels at NaOH concen- 0.20
Fouling rate (kPa/d)

trations below 20 mmol/L, while declined sharply at a concentra- 2.5

tion of 50 mmol/L. Similar with the ex-situ tests results (Fig. 2
0.15
(b)), coenzyme F420 was more sensitive to NaOH while dehydroge- 2.0
nase had the ability to adapt to caustic conditions. The difference is
that the anaerobic biomass of in-situ cleaning bioreactors could 1.5
0.10
tolerate much higher NaOH concentrations than that of ex-situ
cleaning tests, which was due to the limited infiltration of high- 1.0
strength alkali and also the buffering role of the mixed liquor 0.05
0.5
(Fig. S2).
In Fig. 5(b), SMA and BMP were kept at stable levels (23.3 mL
0.0 0.00
CH4/(gVSS
d) and 273.6 mL CH4/gVSS on average, respectively) at
0 10 20 30 40 50
NaOH dosages below 10 mmol/L. With further increase in NaOH NaOH dosage (mmol/L)
dosage, SMA improved slightly and peaked at a NaOH dosage of
20 mmol/L, and thereafter showed inhibition at a extremely high Fig. 6. Economic analysis of the six phases.
 X. Mei et al. / Bioresource Technology 240 (2017) 25–32
cost increased linearly with the increase of NaOH dosages, while
the membrane fouling rate decreased dramatically with enhancing
cleaning strength. Under the comprehensive consideration,
12 mmol/L of NaOH was the cost-efficient and optimal fouling-
control dosage.
3.4. Implications and research needs
In this study, alkali-assisted cleaning with a series of NaOH
dosages was explored for membrane fouling control in AnMBRs
using ceramic membranes, which are resistant to caustic cleaning
solution. The use of NaOH is not only effective in fouling control
but helpful in anaerobic treatment performance. As a preliminary
investigation of CEB cleaning for the operation optimization of
AnMBRs, several issues are worth to be investigated in the future.
Firstly, a relatively higher membrane flux should be operated with
the suggested maintenance cleaning protocol because the current
flux used in this study is low compared to that typically adopted
for aerobic MBRs (i.e., in the range of 20–25 L/(m2
h)). Membrane
permeability is associated to membrane characteristics, biomass
properties, membrane surface shear force and chemical cleaning
strength. The relationship between the membrane permeability
and cleaning strength should be clarified to maximize operating
flux and minimize fouling rate. Secondly, it is of great importance
to study the long-term (e.g., >three year) impact of chemicals on
membrane characteristics and microbial communities. In this
study, ceramic membrane was used because of its high resistance
to high pH. However, polymeric membrane pores were found to be
enlarged during the NaOH backflushing (Zhou et al., 2014), which
may accumulate more irreversible foulants in membrane pores.
Therefore, membrane morphology and long-term changes of
anaerobic microorganisms should be fully understood. Finally, as
our study was based on lab-scale reactors, the maintenance clean-
ing parameters should be further optimized or modified through
pilot-scale AnMBR studies.
4. Conclusions
The impacts of NaOH cleaning on membrane fouling control
and anaerobic microorganisms activities were studied in AnMBRs.
Under short-term explosure to NaOH, NaOH dosages ranging from
0.05 to 1.30 mmol/L had positive effects on anaerobic biomass,
while high dosages (>1.30 mmol/L) had inhibited and/or toxic
effects. In-situ cleaning tests indicated that membrane fouling
rates were significantly reduced by 10–20 mmo/L of NaOH back-
flushing (4–5.5 times over the AnMBR with DI water backflush)
and methanogenic activities were slightly improved. COD removal
efficiencies were over 87% and peaked at 20 mmol-NaOH/L. Eco-
nomic analysis indicates that 12 mmol/L of NaOH was the cost-
efficient and optimal fouling-control dosage.
Acknowledgements
The authors thank the financial support from National Univer-
sity of Singapore, National Science Foundation of China (Grant
51678423), China Scholarship Council (CSC) and Shanghai Tongji
Gao Tingyao Environmental Science and Technology Development
Foundation (STGEF).
Appendix A. Supplementary data
Supplementary data associated with this article can be found, in
the online version, at http://dx.doi.org/10.1016/j.biortech.2017.02.
052.
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References
Ang, W.S., Yip, N.Y., Tiraferri, A., Elimelech, M., 2011. Chemical cleaning of RO
membranes fouled by wastewater effluent: achieving higher efficiency with
dual-step cleaning. J. Membr. Sci. 382, 100–106.
APHA, 2012. Standard Methods for the Examination of Water and Wastewater.
American Public Health Association/American Water Works Association/Water
Environment Federation, Washington, DC, USA.
Arslankaya, E., 2016. Assessment of filtration performance and applicability of low-
cost ceramic membranes to membrane bioreactor (MBR) for municipal
wastewater treatment. Oxid. Commun. 39, 830–837.
Burdock, T., Brooks, M., Ghaly, A., Dave, D., 2011. Effect of assay conditions on the
measurement of dehydrogenase activity of Streptomyces venezuelae using
triphenyl tetrazolium chloride. Adv. Biosci. Biotechnol. 2, 214.
Castro, H., Queirolo, M., Quevedo, M., Muxi, L., 2002. Preservation methods for the
storage of anaerobic sludges. Biotechnol. Lett. 24, 329–333.
Choo, K.H., Lee, C.H., 1996. Membrane fouling mechanisms in the membrane-
coupled anaerobic bioreactor. Water Res. 30, 1771–1780.
Demirel, B., Scherer, P., 2008. The roles of acetotrophic and hydrogenotrophic
methanogens during anaerobic conversion of biomass to methane: a review.
Rev. Environ. Sci. Bio/Technol. 7, 173–190.
Dolfing, J., Bloemen, W., 1985, 1985. Activity measurements as a tool to characterize
the microbial composition of methanogenic environment. J. Microbiol. Methods
4.
Dolfing, J., Mulder, J.W., 1985. Comparison of methane production rate and
coenzyme F420 content of methanogenic consortia in anaerobic granular
sludge. Appl. Environ. Microbiol. 49, 1142–1145.
Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P., Smith, F., 1956. Colorimetric
method for determination of sugars and related substances. Anal. Chem. 28,
350–356.
Frølund, B., Griebe, T., Nielsen, P., 1995. Enzymatic activity in the activated-sludge
floc matrix. Appl. Microbiol. Biotechnol. 43, 755–761.
Han, X.M., Wang, Z.W., Wang, X.Y., Zheng, X., Ma, J.X., Wu, Z.C., 2016. Microbial
responses to membrane cleaning using sodium hypochlorite in membrane
bioreactors: cell integrity, key enzymes and intracellular reactive oxygen
species. Water Res. 88, 293–300.
Hutschemackers, J., Delafontaine, M., Naveau, H., Nyns, E.J., 1982. An improved
method for the quantitative evaluation of the potential for methane
production of mixed microbial communities in digestion mixed liquors
through the determination of the content in coenzyme F420. Biomass 2,
115–125.
Jeison, D., 2007. Anaerobic Membrane Bioreactors for Wastewater Treatment:
Feasibility and Potential Applications. Wageningen Universiteit (Wageningen
University).
Judd, S., 2010. The MBR Book: Principles and Applications of Membrane Bioreactors
for Water and Wastewater Treatment. Elsevier.
Kardos, L., Palko, G., Olah, J., Barkacs, K., Zaray, G., 2009. Operation control of
anaerobic digesters on the basis of enzyme activity tests. Water Sci. Technol. 60,
957–964.
Le-Clech, P., Chen, V., Fane, T.A.G., 2006. Fouling in membrane bioreactors used in
wastewater treatment. J. Membr. Sci. 284, 17–53.
Lee, E.J., Kwon, J.S., Park, H.S., Ji, W.H., Kim, H.S., Jang, A., 2013a. Influence of sodium
hypochlorite used for chemical enhanced backwashing on biophysical
treatment in MBR. Desalination 316, 104–109.
Lee, E.J., An, A.K.J., Hadi, P., Yan, D.Y.S., 2016. Characterizing flat sheet membrane
resistance fraction of chemically enhanced backflush. Chem. Eng. J. 284, 61–67.
Lee, S.J., Dilaver, M., Park, P.K., Kim, J.H., 2013b. Comparative analysis of fouling
characteristics of ceramic and polymeric microfiltration membranes using
filtration models. J. Membr. Sci. 432, 97–105.
Li, H., Jin, Y., Mahar, R., Wang, Z., Nie, Y., 2008. Effects and model of alkaline waste
activated sludge treatment. Bioresour. Technol. 99, 5140–5144.
Liang, S., Liu, C., Song, L., 2007. Soluble microbial products in membrane bioreactor
operation: behaviors, characteristics, and fouling potential. Water Res. 41, 95–
101.
Lin, J.G., Chang, C.N., Chang, S.C., 1997. Enhancement of anaerobic digestion of waste
activated sludge by alkaline solubilization. Bioresour. Technol. 62, 85–90.
McCarty, P.L., Bae, J., Kim, J., 2011. Domestic wastewater treatment as a net energy
producer-can this be achieved? Environ. Sci. Technol. 45, 7100–7106.
Mei, X.J., Wang, Z.W., Miao, Y., Wu, Z.C., 2016. Recover energy from domestic
wastewater using anaerobic membrane bioreactor: operating parameters
optimization and energy balance analysis. Energy 98, 146–154.
Owen, W.F., Stuckey, D.C., Healy Jr, J.B., Young, L.Y., McCarty, P.L., 1979. Bioassay for
monitoring biochemical methane potential and anaerobic toxicity. Water Res.
13, 485–492.
Ozgun, H., Dereli, R.K., Ersahin, M.E., Kinaci, C., Spanjers, H., van Lier, J.B., 2013. A
review of anaerobic membrane bioreactors for municipal wastewater
treatment: integration options, limitations and expectations. Sep. Purif.
Technol. 118, 89–104.
Ramos, C., Zecchino, F., Ezquerra, D., Diez, V., 2014. Chemical cleaning of
membranes from an anaerobic membrane bioreactor treating food industry
wastewater. J. Membr. Sci. 458, 179–188.
Rosenberger, S., Laabs, C., Lesjean, B., Gnirss, R., Amy, G., Jekel, M., Schrotter, J.C.,
2006. Impact of colloidal and soluble organic material on membrane
performance in membrane bioreactors for municipal wastewater treatment.
Water Res. 40, 710–720.
32 X. Mei et al. / Bioresource Technology 240 (2017) 25–32
Scherson, Y.D., Criddle, C.S., 2014. Recovery of freshwater from wastewater:
upgrading process configurations to maximize energy recovery and minimize
residuals. Environ. Sci. Technol. 48, 8420–8432.
Simon, F.X., Penru, Y., Guastalli, A.R., Esplugas, S., Llorens, J., Baig, S., 2013. NOM
characterization by LC-OCD in a SWRO desalination line. Desalin. Water Treat.
51, 1776–1780.
Smith, A.L., Stadler, L.B., Cao, L., Love, N.G., Raskin, L., Skerlos, S.J., 2014. Navigating
wastewater energy recovery strategies: a life cycle comparison of anaerobic
membrane bioreactor and conventional treatment systems with anaerobic
digestion. Environ. Sci. Technol. 48, 5972–5981.
Stuckey, D.C., 2012. Recent developments in anaerobic membrane reactors.
Bioresour. Technol. 122, 137–148.
Tang, S.J., Wang, Z.W., Wu, Z.C., Zhou, Q., 2010. Role of dissolved organic matters
(DOM) in membrane fouling of membrane bioreactors for municipal
wastewater treatment. J. Hazard. Mater. 178, 377–384.
Tian, J.Y., Chen, Z.L., Yang, Y.L., Liang, H., Nan, J., Li, G.B., 2010. Consecutive chemical
cleaning of fouled PVC membrane using NaOH and ethanol during
ultrafiltration of river water. Water Res. 44, 59–68.
Walczak, M., Cywinska, A., 2007. Application of selected chemical compounds to
limit the growth of filamentous bacteria in activated sludge. Environ. Prot. Eng.
33, 221.
Wang, Z., Ma, J., Tang, C.Y., Kimura, K., Wang, Q., Han, X., 2014a. Membrane cleaning
in membrane bioreactors: a review. J. Membr. Sci. 468, 276–307.
Wang, Z., Meng, F., He, X., Zhou, Z., Huang, L.N., Liang, S., 2014b. Optimisation and
performance of NaClO-assisted maintenance cleaning for fouling control in
membrane bioreactors. Water Res. 53, 1–11.
Wang, Z.W., Ma, J.X., Tang, C.Y.Y., Kimura, K., Wang, Q.Y., Han, X.M., 2014c.
Membrane cleaning in membrane bioreactors: a review. J. Membr. Sci. 468,
276–307.
Wei, C.H., Huang, X., Ben Aim, R., Yamamoto, K., Amy, G., 2011. Critical flux and
chemical cleaning-in-place during the long-term operation of a pilot-scale
submerged membrane bioreactor for municipal wastewater treatment. Water
Res. 45, 863–871.
Yu, H., Wang, Z., Wang, Q., Wu, Z., Ma, J., 2013a. Disintegration and acidification of
MBR sludge under alkaline conditions. Chem. Eng. J. 231, 206–213.
Yu, H.G., Wang, Z.W., Wang, Q.Y., Wu, Z.C., Ma, J.X., 2013b. Disintegration and
acidification of MBR sludge under alkaline conditions. Chem. Eng. J. 231, 206–
213.
Yue, X.D., Koh, Y.K.K., Ng, H.Y., 2015. Effects of dissolved organic matters (DOMs) on
membrane fouling in anaerobic ceramic membrane bioreactors (AnCMBRs)
treating domestic wastewater. Water Res. 86, 96–107.
Zabranska, J., Schneiderova, K., Dohanyos, M., 1985. Relation of coenzyme F420 to
the activity of methanogenic microorganisms. Biotechnol. Lett. 7, 547–552.
Zhou, Z., Meng, F., Lu, H., Li, Y., Jia, X., He, X., 2014. Simultaneous alkali
supplementation and fouling mitigation in membrane bioreactors by on-line
NaOH backwashing. J. Membr. Sci. 457, 120–127.