Article

pubs.acs.org/est

Electrochemical Nutrient Recovery Enables Ammonia Toxicity

Control and Biogas Desulfurization in Anaerobic Digestion
Joachim Desloover, Jo De Vrieze, Maarten Van de Vijver, Jacky Mortelmans, Rene Rozendal,
and Korneel Rabaey*
Laboratory of Microbial Ecology and Technology (LabMET), Ghent University, Coupure Links 653, B-9000 Gent, Belgium
S Supporting Information
*

ABSTRACT: Organic waste streams can be valorized and reduced in volume

with anaerobic digestion (AD). An often-encountered key issue however is the
high ammonium (NH4+) content of certain waste streams. Ammonia (NH3), in
equilibrium with NH4+, is a toxic compound to the methanogenic community,
which limits the organic loading rate and endangers process stability. An
electrochemical system (ES) linked to a digester could, besides recovering this
nutrient, decrease NH3 toxicity through electrochemical extraction. Therefore,
two digesters with and without ES attached in the recirculation loop were
operated to test whether the ES could control NH3 toxicity. During periods of
high ammonium loading rates, the methane (CH4) production of the ES-coupled
reactor was up to 4.5 times higher compared to the control, which could be
explained through simultaneous NH4+ extraction and electrochemical pH control.
A nitrogen ux of 47 g N m2 membrane d1 could be obtained in the ES-coupled reactor, resulting in a current and removal
eciency of 38 5% and 28 2%, respectively, at an electrochemical power input of 17 2 kWh kg1 N. The anode also
oxidized sulde, resulting in a signicantly lower H2S emission via the biogas. Lastly, limited methanogenic community dynamics
pointed to a nonselective inuence of the dierent operational conditions.

INTRODUCTION
Anaerobic digestion (AD) is a key technology for stabilization
and valorization of organic waste streams.1,2 In short, this
technology comprises a stepwise conversion of low-value
organic compounds into biogas, a mixture of mainly methane
(CH4) and carbon dioxide (CO2). Methane is an energy carrier
and can be valorized through, for example, a combined heat and
power unit, delivering electricity and heat. Next to biogas, AD
also produces a nutrient-rich digestate that can be applied as a
fertilizer in agriculture.3 Despite numerous advantages of the
AD process, instability is an often-encountered problem that
can lead to complete failure of the reactor. One of the key
compounds causing instability is ammonia (NH3), especially
when treating nitrogen-rich waste streams.4,5 Ammonia is a cell
membrane-permeable molecule for which methanogens,
executing the nal step in AD, have a low tolerance.6 Moreover,
acetoclastic methanogenesis is, in general, more susceptible to
inhibition than hydrogenotrophic methanogenesis.6 This
exposes digesters to a risk of process instability and limits the
loading and thus biogas production rate.4 To avoid this,
operators usually feed the digester at a lower and safer loading
rate, acclimate the biomass, or co-digest with carbon-rich
substrates to maintain a suitable carbon to nitrogen ratio.5
Other more advanced but also more expensive approaches are
struvite precipitation, anammox, and the use of zeolites.5
In this study, we present an alternative to control NH3
toxicity and to maximize resource recovery from AD by
coupling an electrochemical system (ES) to an anaerobic
XXXX American Chemical Society

digester. An ES has the attractive feature that an oxidation

process (anode) is separated from a reduction process
(cathode), typically by an ion selective membrane. By applying
a current to this system, membrane electrolysis can take place
during which ions can be extracted from anode to cathode or
vice versa.7,8 In the context of anaerobic digestion, one can send
the digestate through an anode compartment, enabling
recovery of valuable nutrients such as ammonium (NH4+)
and potassium (K+). Hence, by combining AD with ES
technology, nutrients can be harvested while simultaneously
lowering the risk for ammonia toxicity. We have recently
demonstrated the proof of concept of an ES for nutrient
recovery from liquid waste streams.8
Here, we studied the direct coupling of an ES to an upow
anaerobic sludge blanket (UASB) reactor treating molasses. We
investigated whether the placement of an anode in the
recirculation line had any negative eects on the digester and
whether the ES could stabilize AD performance when exposed
to toxic NH3 concentrations. Lastly, we also investigated in
what manner the ES can aect the quality of the biogas
generated.
Received: October 1, 2014
Revised: December 13, 2014
Accepted: December 17, 2014

DOI: 10.1021/es504811a
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MATERIAL AND METHODS

Experimental Setup. Two cylindrical UASB reactors (2.3
L glass reactor with eective volume of 2 L) were constructed,
serving as test and control reactors (Figure 1). These reactors

Table 1. Composition Tap Water-Diluted Molasses To

Obtain Desired Loading Rate of 5 g COD L1 d1.
parameter

value

unit

pH
conductivity
COD
Kj-N
TAN
SO42
T-P
Cl
TS
VS
VSS

5.7 0.3
8.5 1
11.7 2.2
439 19
16
254
80
156
11
9
0.8

mS cm1
g L1
mg L1
mg N L1
mg L1
mg P L1
mg L1
g L1
g L1
g L1

UASBs to compensate for any later addition of NH4Cl, when

the performance was investigated under high nitrogen loading
conditions. Both reactors were pH controlled (Dulcometer
D1C, Prominent, Germany) with 1 M NaOH. The electrochemical process parameters of the ES were dened and
calculated according to Desloover et al.8
The cathode compartments of the control and test setups
were fed continuously with 6.4 g L1 NaCl at 1 L d1 (HRT of
4.8 h) with an internal recirculation rate of 2 L h1.
Experimental Plan. The experimental plan comprised four
main phases (Table 2). During Phase I, the organic loading rate
was gradually increased from 1 to 5 g COD L1 d1 (Phase Ia).
After stable operation, the pH was stepwise (0.25 pH units per
week) increased from 7 to 8 (Phase Ib) to shift the NH4+/NH3
equilibrium more to the direction of NH3 (ratio NH3/NH4+ =
0.11 at pH 8 and 34 C). The free ammonia fraction was
calculated according to Anthonisen et al.10 Next, the ES of the
test setup was switched on at an applied current density of 10 A
m2 (relative to projected membrane surface area) to
investigate the impact during low nitrogen loading. At day
120, the UASB of the test setup crashed due to clogging and
subsequent malfunction of the pH controller. Hence, both the
test and control reactors were cleaned, and the biomass of the
control UASB was split over the test and control setups to
initiate a second start-up (Phase IIIa) during which the organic
loading rate and pH were maintained at 5 g COD L1 d1 and
8, respectively. Also, the ES of the test setup was stepwise
increased from 5 to 10 A m2. After a steady-state period
(Phase IIIb), the eect of the ES was investigated under
periodically increased nitrogen loading conditions (Phase IV).
Therefore, the eect of an operational ES on the test setup was
investigated during a period of increased nitrogen loading
(Phase IVa), as well as a period during which the extra-added
nitrogen was again removed from the feed (Phase IVb). Next,
this operational procedure was repeated during a period where
the ES of the test setup was switched o (Phases IVc and IVd).
Finally, after an adaptation period where the ES was switched
on again (Phase IVe), the eect of the ES was investigated
during additional nitrogen loading up to 2 g N L1 (Phases IVf
and IVg) and where we also allowed the electrochemical cell to
control the pH by taking advantage of the acidifying anode
reaction.
Chemical Analysis. Liquid samples of the inuent and
euent streams as well as gaseous samples from the headspace
were taken three times a week. Liquid samples were ltered
(0.22 m) and stored at 4 C until further analysis.

Figure 1. Schematical overview of the experimental setup.

had an internal diameter of 5.4 cm and a total height of 900 cm.

For the test reactor, an ES was coupled to the UASB for
extraction of cations. This was done by inserting the anode
compartment (5 cm 20 cm 2 cm) of the ES in the
recirculation loop of the UASB reactor. The anode compartment was separated from the cathode compartment (5 cm 20
cm 2 cm3) by a cation exchange membrane (CEM,
Membranes International, U.S.A.). The anode electrode used
was an IrOx coated titanium mesh electrode 9 (12g m2 of Ir/
Ta = 65/35), with a projected surface area of 5 cm 20 cm
(Magneto Special Anodes, The Netherlands), while the
cathode electrode was a stainless steel mesh (5 20 cm2,
mesh width 564 m Solana, Belgium). Both electrodes were
placed close to the CEM and separated by a polytetrauoroethylene (PTFE) spacer with a projected surface area of 5 cm
20 cm (turbulence promoter mesh, Electrocell, Denmark) to
avoid direct contact. At the anode, water was oxidized to
oxygen and protons, while at the cathode, water was reduced to
hydrogen gas and hydroxyl ions. The ES was controlled
galvanostatically by a VSP multipotentiostat (Biologic, France).
The control reactor was also coupled to an ES in the
recirculation loop. The ES was equipped with a CEM, but
electrodes were omitted. Hence, the ES of the control setup
was operated in open circuit (no anode and cathode), meaning
that no current could be applied to the system and only
diusion driven processes could take place.
Reactor Operation. The experiment was conducted under
mesophilic conditions (34 1 C). The UASB reactors were
inoculated with granular sludge from a full-scale UASB reactor
(Brewery Van Steenberge, Belgium) and diluted with tap water
to obtain an initial sludge concentration of 10 g of volatile
suspended solids (VSS) L1. The UASBs were fed every two
hours with tap water-diluted molasses according to the desired
loading rate and were operated at a hydraulic retention time
(HRT) of 2 days. The characteristics of the diluted molasses to
obtain a desired loading rate of 5 g COD L1 d1 are shown in
Table 1 (raw composition molasses, Table S1, Supporting
Information). Furthermore, an internal recirculation rate was
applied over the UASB and anode compartment of 2 L h1 to
maintain an upow velocity of 1 m h1 in the digester. In order
to maintain the same conductivity throughout the experimental
period, 4.14 g L1 NaCl was initially added to the feed of both
B

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Table 2. Overview of Experimental Plan
phase

operation

period (d)

Ia
Ib
II
IIIa
IIIb
IVa
IVb
IVc
IVd
IVe
IVf
IVg

start-up 1
gradual increase pH 78
switch on ES test setup
start-up 2 + switch on ES test setup
steady state
add 1 g N L1 to feed
remove additional 1 g N L1 from feed
switch o ES test setup + add 1 g N L1 to feed
remove additional 1 g N L1 from feed
switch on ES test setup
add 1 g N L1 to feed
add 2 g N L1 to feed + electrochemical acidication (pH 87)

130
3070
70120
130210
210225
225238
238266
266275
275287
287303
303313
313340

dierent parameters obtained during analysis with the

StepOnePlus software V2.3 (Table S2, Supporting Information).
Statistical Analysis. All statistical data analysis were
performed with the statistical software R, version 3.0.2. for
Windows. In the case of normally distributed data sets that
were homoscedastic the regular t test was applied. In the case
where the data was heteroscedastic, a Welch-modied t test was
used. In the case where the data was not normally distributed,
the Wilcoxon Rank Sum (MannWhitney U) test was applied.

Volatile suspended solids (VSS), Kjeldahl nitrogen (Kj-N),

ammonium (NH4+), chemical oxygen demand (COD), pH,
and conductivity were analyzed according to standard
methods.11 Volatile fatty acids (VFA) were, after extraction in
diethyl ether, analyzed with a DB-FFAP 12-3232 column (30 m
0.32 mm 0.25 m; Agilent, Belgium) and a ame
ionization detector (FID) gas chromatograph (GC-2014,
Shimadzu, The Netherlands). The gas phase composition was
analyzed with a compact GC (Global Analyzer Solutions,
Breda, The Netherlands). The GC was equipped with two
channels. In channel 1, a Porabond precolumn and Molsieve 5A
column were used for CH4, O2, H2, and N2 measurement, and
in channel 2 a Rt-Q-bond precolumn and column were used for
CO2, N2O, and H2S analysis. Concentrations of gases were
determined by means of a thermal conductivity detector and
were reported at STP (standard temperature and pressure)
conditions. Biogas production was measured with an in-house
manufactured calibrated gas counter.
Molecular Analysis. Total DNA was extracted from the
sludge samples using the protocol of Vilchez-Vargas et al.12
DNA quality and quantity of the extracts was analyzed by
means of a 1% agarose gel and a Nanodrop ND-1000
spectrophotometer (Isogen Life Science, IJsselstein, The
Netherlands). Triplicate samples of a 100-fold dilution of the
DNA samples were prepared to reach a nal DNA
concentration between 1 and 10 ng L1.
Real-time PCR (qPCR) was performed on a StepOnePlus
Real-Time PCR System (Applied Biosystems, Carlsbad, CA,
U.S.A.). The reaction mixture of 15 L was prepared using the
GoTaq qPCR Master Mix (Promega, Madison, WI, U.S.A.) and
consisted of 10 L of GoTaq PCR Master Mix, 3.5 L of
nuclease-free water, and 0.75 L of each primer (nal
concentration of 375 nM), to which 5 L of template DNA
was added. The qPCR program was performed in a two-step
thermal cycling procedure that consisted of a predenaturation
step of 10 min at 94 C, followed by 40 cycles of 15 s at 94 C
and 1 min at 60 C for total bacteria, using the P338F and
P518r primers, as described by Ovreas et al.13 The qPCR
program for the methanogenic orders Methanobacteriales and
the families Methanosaetaceae and Methanosarcinaceae consisted
of a predenaturation step of 10 min at 94 C, followed by 40
cycles of 10 s at 94 C and 1 min at 60 C. For quantication of
the Methanomicrobiales order an annealing temperature of 63
C was used. The primers for the methanogenic orders
Methanomicrobiales and Methanobacteriales and the families
Methanosaetaceae and Methanosarcinaceae were described by Yu
et al.14 Real-time PCR quality was evaluated by means of the

RESULTS AND DISCUSSION

ES Has a Temporal Eect on Digester during Low
Nitrogen Loading Conditions. After the start-up phase
(Phase Ia), the average CH4 production rate of the test and
control reactors during Phase Ib was 949 90 and 950 134
mL CH4 L1 d1, respectively (Figure 2, Table S3, Supporting

Figure 2. CH4 production in function of time of the test and control

setup during Phases IaII.

Information). Hence, gradually adapting the pH from 7 to 8 did

not have a notable eect on methane production. Moreover, by
operating at a high pH, a CH4 content up to 83% could be
reached in both the test and control reactors (Table S3,
Supporting Information).
When the ES of the test setup was switched on at the start of
Phase II, an initial decrease in CH4 production rate of about
C

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Figure 3. CH4 production (A), VFA concentration (B), NH4+ concentration (C), and H2S content in the biogas (D) in function of time of the test
and control setups during Phases IIIaIVg. The labels presented in this gure account for all graphs.

performance (Figure 3A). Furthermore, the decline in CH4

production of the control reactor coincided with the
accumulation of VFA up to 2700 mg COD L1, whereas no
VFA could be detected in the test reactor (Figure 3B). VFA
accumulation is a strong sign of methanogen inhibition, and as
described in other studies, this was most probably caused by the
high ammonium content in combination with a high pH.6,15
The fact that the test reactor outperforms the control reactor
can thus be explained by an on average 23% lower ammonium
level in the test reactor caused by membrane electrolysis
(Figure 3C, Table 3). By omitting the additional nitrogen from
the feed (Phase IVb), the control reactor was able to partially
recover over a period of 30 days (Figure 3A).
Repeating this procedure with a nonworking ES of the test
reactor (Phases IVc and IVd) resulted in a decrease in
performance of both the test and control reactors (Figure 3A).
The ammonium levels in both reactors were identical (Figure
3C), and also, this time not only VFA accumulation could be
observed in the control reactor, but also in the test reactor
(Figure 3B). These ndings prove that ammonium extraction
by the ES was essential to maintain constant increased methane
production values under high nitrogen loading conditions.
Most likely, the corresponding average NH3 concentration
during Phase IVa in the test reactor (94 mg N L1) did not
reach a level that inhibited the methanogenic community,

20% could be observed. The temporary negative impact on the

performance of the microbial community was probably due to a
shock eect caused by the instantaneous oxygen and proton
production by the anode reaction, in combination with a higher
NaOH dosage to counteract acidication (Table S3, Supporting
Information). The produced oxygen represented 7% of the
COD loading rate at 10 A m2 but was consumed because no
O2 could be detected in the biogas. Moreover, the conductivity
of the test (19.3 2.7 mS cm1) and control (21.1 3.2 mS
cm1) reactors were not signicantly dierent (Table S3,
Supporting Information, p > 0.05), due to membrane
electrolysis.
After 40 days of operation, the test reactor recovered and
reached again the performance of the control reactor (Figure
2).
Electrochemical NH3 Toxicity Control during High
Nitrogen Loading Conditions. After the crash of the test
reactor and a second start-up (Phase IIIa), both reactors again
reached equal performance (Phase IIIb, p > 0.05), with a CH4
production rate of 925 94 and 848 66 mL CH4 L1 d1 in
the test and control reactors, respectively (Figure 3A, Table 3).
When the nitrogen loading was increased by adding an
additional 1 g N L1 to the feed (Phase IVa), a 43% decrease in
the CH4 production rate could be observed for the control
reactor, while the test reactor was able to maintain its
D

DOI: 10.1021/es504811a
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Table 3. Overview of Parameters in Test and Control Setups during Phases IIIaIVca
parameter
UASB (with anode
compartment)
CH4 production
(mL CH4 L1d1)
eciencyb (%)
sCOD euent
(g COD L1)
total VFA
(mg COD L1)
acetate (mg L1)
propionate (mg L1)
TAN (mg N L1)
conductivity (mS cm1)
pH ()
NaOH dosage (mL d1)
CH4 (%)
H2S (%)
Cathode
TAN (mg N L1)
conductivity (mS cm1)
pH ()
Electrochemical
N uxc (g N m2 d1)
NH4+ current eciency
(CE,%)
NH4+ removal eciency
(RE,%)
cell voltage (V)
energy input
(kWh kg1 N)

Phase IIIa (n = 27)

Phase IIIb (n = 6)

Phase Iva (n = 6)

Phase IVb (n = 12)

Phase IVc (n = 4)

test

control

test

control

test

control

test

control

test

control

618 173

626 155

925 94

848 66

890 51

571 121

940 64

740 63

749 147

551 107

36 10
ND

36 9
ND

52 2
0.96 0.26

50 2
2.04 0.37

54 3
0.97 0.02

34 8
2.82 0.40

56 5
0.93 0.12

44 4
3.71 1.21

45 10
3.30 0.99

34 7
4.45 0.75

ND

ND

BDL

630 306

BDL

1939 704

BDL

1983 429

1575 817

3119 730

ND
ND
ND
20.1 2.1
7.9 0.1
106 30
82 4
0.25 0.15

ND
ND
ND
19.7 2.8
7.9 0.1
81 10
84 3
0.27 0.06

BDL
BDL
250 30
16.0 0.3
7.9 0.1
142 28
92 2
0.05 0.07

249 95
370 203
294 20
17.5 0.5
8.0 0.1
73 5
94 1
0.22 0.03

BDL
BDL
823 116
18.3 1.3
7.9 0.1
166 16
89 1
BDL

1218 459
670 259
1069 51
17.8 0.5
8.0 0.1
83 4
93 1
0.22 0.04

BDL
BDL
302 54
18.6 1.9
8.0 0.2
155 15
91 7
0.05 0.08

1299 293
609 163
376 69
20.0 1.3
7.9 0.1
80 12
91 3
0.19 0.09

1133 645
408 147
1001 199
22.3 1.3
8.0 0.1
95 17
89 1
0.51 0.10

2519 669
510 71
1001 195
21.6 1.2
8.0 0.1
86 10
89 2
0.23 0.01

ND
17.0 5.2
9.9 2.2

ND
14.1 3.8
7.9 0.3

42 21
22.4 5.5
12.5 0.1

10 6
9.4 2.7
7.8 0.2

220 20
20.6 2.0
12.1 0.1

38 12
12.0 1.1
7.6 0.1

60 15
24.0 3.3
12.4 0.1

23 8
12.2 1.4
7.7 0.2

24 11
12.3 1.5
8.5 0.3

44 9
12.6 2.3
7.9 0.1

ND
ND

ND
ND

42
31

11
NA

19 2
15 1

31
NA

51
41

21
NA

21
NA

30
NA

ND

ND

14 6

32

22 3

31

17 1

62

31

40

ND
ND

ND
ND

3.58 0.65
307 182

NA
NA

4.72 0.91
60 6

NA
NA

4.72 0.91
251 37

NA
NA

NA
NA

NA
NA

a
n = number of data points. ND: not determined. BDL: below detection limit. NA: not applicable. bCOD to CH4 conversion eciency at STP
conditions and relative to the amount of COD fed to the reactor. cRelative to the projected membrane surface area.

Figure 4. Relative abundance of methanogens (Methanomicrobiales, Methanobacteriales, Methanosarcinacea, and Methanosaetaceae) in the test (T) and
control (C) reactors throughout the experimental period.

instantaneous recovery of the test reactor, while the control

reactor seemed to reach an inhibited steady state,4 which was
on average 43% lower in performance compared to the test
reactor.
Gradually switching on the ES of the test reactor from 5 to
10 A m2 caused again a temporary negative eect, comparable

whereas inhibition might have occurred in the control reactor

were a higher average NH3 concentration was present (118 mg
N L1). Indeed, reported inhibitory NH3 concentrations are
within a range of 80150 mg N L1 and are dependent on the
operational conditions and degree of adaptation.4,5 Removal of
the extra-added NH4+ from the feed (Phase IVd) caused
E

DOI: 10.1021/es504811a
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Table 4. Overview of Parameters in Test and Control Setups during Phases IVdIVga
parameter
UASB (with anode compartment)
CH4 production (mL CH4 L1d1)
eciencyb (%)
sCOD euent (g COD L1)
total VFA (mg COD L1)
acetate (mg L1)
propionate (mg L1)
TAN (mg N L1)
conductivity (mS cm1)
pH ()
NaOH dosage (mL d1)
CH4 (%)
H2S (%)
Cathode
TAN (mg N L1)
conductivity (mS cm1)
pH ()
Electrochemical
N uxc (g N m2 d1)
NH4+ current eciency (CE,%)
NH4+ removal eciency (RE,%)
cell voltage (V)
energy input (kWh kg1 N)

Phase IVd (n = 5)

Phase IVe (n = 7)

Phase IVf (n = 4)

Phase IVg (n = 12)

test

control

test

control

test

control

test

control

973 87
58 8
2.1 0.7
780 529
504 326
260 180
504 128
21.7 0.6
8.1 0.1
72 15
89 0
0.27 0.10

537 39
33 4
4.7 0.2
3239 281
2611 237
545 51
482 121
23.6 0.9
8.0 0.1
82 5
90 1
0.22 0.02

798 136
50 9
2.7 0.8
1019 386
840 291
86 43
284 16
21.5 2.6
8.0 0.1
140 43
89 2
0.11 0.03

507 33
32 3
4.8 0.3
3735 210
2834 140
442 84
341 26
24.4 2.3
8.0 0.1
94 24
91 1
0.19 0.03

786 139
48 7
3.5 1.4
3052 1493
2465 1096
233 182
703 198
17.7 0.9
8.0 0.1
179 26
93 2
0.08 0.02

458 22
30 3
5.7 0.8
3291 686
2552 528
312 96
1040 120
21.3 3.6
7.9 0.1
88 11
90 1
0.18 0.02

644 188
40 11
3.1 1.0
1459 1041
1204 876
78 53
1527 92
23.6 1.8
7.1 0.2
109 39
64 11
0.12 0.13

252 87
16 5
7.0 0.4
6252 1000
5168 878
440 35
1889 113
27.0 2.1
7.9 0.1
82 5
90 3
0.19 0.02

28 5
12.1 0.3
8.3 0.2

44 8
13.6 3.2
7.9 0.2

109 15
30.1 3.4
12.6 0.2

45 9
18.6 4.5
8.1 0.2

296 51
25.6 4.3
12.5 0.3

99 15
16.6 5.0
7.8 0.2

600 101
18.6 2.6
12.4 0.2

155 19
14.1 1.3
7.9 0.1

20
NA
51
NA
NA

31
NA
81
NA
NA

91
92
29 5
1.69 0.13
39 9

31
NA
11 3
NA
NA

23 3
18 2
27 4
3.40 0.25
36 4

72
NA
92
NA
NA

45 8
36 6
27 3
3.25 0.10
18 3

13 1
NA
81
NA
NA

a
n = number of data points. ND: not determined. BDL: below detection limit. NA: not applicable. bCOD to CH4 conversion eciency at STP
conditions and relative to the amount of COD fed to the reactor. cRelative to the projected membrane surface area.

to Phase II. However, this time the recovery took only 16 days
(Phase IVe). This shows that gradual increase of the current
density is necessary to allow adaptation of the microbial
community to the new conditions. The limited degree of
change in the methanogenic community (Figure 4) indicates
that adaptation of the microbial community took place through
physiological responses rather than microbial community
composition variation.
During the nal two phases (Phases IVf and IVg), the
nitrogen loading was again increased by addition of 1 (Phase
IVf) and 2 (Phase IVg) g N L1 to the feed. In contrast to
Phase IVa, the CH4 production rate of the test reactor started
to decrease dramatically (Figure 3A), which can be explained
by the fact that the methanogenic community was also stressed
in the test reactor because residual VFA were present (Figure
3B). A further increase in the nitrogen loading (additional 2 g
N L1) led to a minimum CH4 production rate of 322 mL of
CH4 L1 d1 at day 317, which was equal to the performance of
the control reactor (Figure 3A). Clearly, the extraction of
ammonium by the ES was insucient to decrease the NH4+
and hence also the NH3 concentration below a toxic level
(Figure 3C).
Therefore, from day 317 onward, not only the extraction
capacity of the ES was utilized but also the ability to acidify and
hence control the pH of the reactor through the use of the
acidifying anode oxidation reaction. Until day 317, the
generated protons by the ES were counteracted by NaOH
addition in order to operate the test and control setups at the
same pH. As a result, NaOH addition to the test reactor was
almost double of the control reactor whenever the ES was
switched on (Tables 3 and 4). This, however, did not generate
a signicant (p > 0.05) dierence in conductivity between both
setups, as the base addition to the test reactor was compensated

by membrane extraction of the ES. By steadily decreasing the

base dosage of the test reactor down to the level of the control
reactor, the pH of the test reactor evolved to 7.1 (Table 4), and
as such, the acidifying eect of the ES could be investigated.
The test reactor completely recovered and reached a CH4
production rate of 856 38 mL of CH4 L1 d1 during the last
seven days of operation, while the control reactor remained at a
4.5 times lower CH4 production rate of 192 10 mL of CH4
L1 d1. This coincided with a steep decrease in VFA below
detection limit in the test reactor and VFA accumulation up to
7500 mg L1 in the control reactor. From this, we can conclude
that, next to electrochemical NH4+ extraction, also electrochemical pH control is a powerful tool allowing ecient NH3
toxicity control in this study. Moreover, electrochemical NH4+
extraction generates added value as it allows for recovery of this
nutrient here under the form of a H2/NH3 gas mixture. This
gas mixture can easily be separated via, for example, ammonia
condensation, thus delivering a concentrated liquid ammonium
stream and a puried H2 gas stream for injection in the
anaerobic digester.
Electrochemical Nutrient Recovery. The concept of
electrochemical nutrient recovery has been demonstrated
previously, both with an electrochemical 8 as with a
bioelectrochemical system,16,17 whether or not in combination
with power consumption or production. This concept was also
applied in this study, especially during the nal phase (Phase
IVg). In this phase, the need for a high ammonium
concentration in order to obtain a high extraction eciency
was again shown (Figure 4C). Here, NH4+ could be extracted at
a ux of 47 6 g N m2 membrane d1, resulting in a removal
and current eciency of 36 6% and 27 3%, respectively
(Table 4). In terms of electrochemical power input, the
ammonium could be extracted at 17 2 kWh kg1 N. This is
F

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Environmental Science & Technology

the main acetoclastic methanogens relates to their lamentous

structure and therefore crucial role in anaerobic granule
formation.24 The persistence of the Methanobacteriales and
Methanomicrobiales order points out that not only acetoclastic
but also hydrogenotrophic methanogenesis, whether or not in
combination with syntrophic acetate oxidation, could take
place. The overall limited dynamics of the methanogenic
community composition and abundance indicates that the
methanogenic community was not selectively inuenced by the
operational conditions in the rapid succession of the dierent
phases.

comparable to the results obtained in our previous study where

we obtained a power input of 13 kWh kg1 N at a current
density of 10 A m2 and 2 g N L1 with digestate from a
municipal solid waste digester.8 Simultaneously with the NH3,
K+ is removed from the digestate, and upon extraction of the
latter, the energy input could be spread over those two
products. Ammonium is driven to a solids-free catholyte with
high pH (Tables 3 and 4) and converted into volatile NH3. The
NH3 can then be recovered from this stream through stripping
and absorption technology8 but was not subject of this study.
Extra Asset: Electrochemical Remediation of H2S. An
interesting side observation during this study was the
signicantly lower H2S content in the biogas of the test reactor
when the ES was switched on (Figure 3D, p < 0.05). Under
these conditions, the H2S concentration in the biogas of the test
reactor was often below the detection limit (0.01%, Tables 3
and 4), except for the period after the crash and cleaning of the
reactors (Phase IIIa) when the ES was switched on. However, a
lot of variation was observed due to the dierent phases. As
such, H2S was higher during the period after the crash and
cleaning of the reactors (Phase IIIa) and at the beginning of
Phase IVg, when the pH shifted from 8 to 7. In contrast, the
H2S concentration in the control setup was on average 0.20
0.06% over the entire experimental period. The overall lower
H2S content in the biogas of the test reactor when the ES was
switched on was most likely caused by direct or indirect
electrochemical oxidation of the dissolved sulde species in the
vicinity of the anode electrode. Electrochemical sulde removal
from domestic wastewater was recently studied in detail at Ir/
Ta-coated MMO-coated titanium anodes.18 The main mechanism was indirect sulde oxidation to elemental sulfur,
thiosulphate, and sulfate by the in situ produced oxygen at
the anode. Most likely, this also took place in our setup, as a
similar anode electrode was used. Next to plain electrochemical
oxidation, also bioelectrochemical oxidation could take place as
microorganisms were growing in the anode compartment.19 A
yellow deposition was observed on the anode, indicating the
production of elemental sulfur. However, due to the complex
mixed liquor, it was not possible to analyze this.
The presence of H2S constitutes a severe problem to any
biogas conversion technology as it can cause corrosion.20
Hence, electrochemical H2S removal is a valuable asset next to
NH3 toxicity control and nutrient recovery. In combination
with a typically high reactor pH, the biogas has a low CO2
content as well, leading to a highly methane enriched biogas.
Microbial Community Findings. Microbial community
analysis was carried out to evaluate whether a dierentiating
impact could be observed due to the presence of the ES and
through time. Real-time PCR analysis of the total bacteria and
summation of the dierent methanogenic groups resulted in an
overall coverage of the microbial community by the
methanogens between 1% and 5% only (Figure S1, Supporting
Information). This is in contrast to their crucial role in the
anaerobic digestion process and anticipated activity, yet is in
agreement with other lab- and full-scale AD installations in
which the methanogenic community covered no more than 5%
of the total microbial community.2123 Analysis of the main
dierent methanogenic populations in AD demonstrated the
presence of Methanobacteriales, Methanomicrobiales, and Methanosaetaceae for at least 10% of the methanogenic community
in each sample, irrespective of the treatment or time point
(Figure 4). The assumed dominance of Methanosaetaceae over
Methanosarcinaceae (below detection limit in every sample) as

ASSOCIATED CONTENT

S Supporting Information
*

Information as mentioned in the text. This material is available

free of charge via the Internet at http://pubs.acs.org.

AUTHOR INFORMATION

Corresponding Author

*Phone: +32 (0)9 264 59 76. Fax: +32 (0)9 264 62 48. E-mail:
Korneel.Rabaey@ugent.be. Web page: www.labmet.Ugent.be.
Notes

The authors declare no competing nancial interest.

ACKNOWLEDGMENTS
J.D. is supported by an Advanced Grant of the Industrial
Research Fund at Ghent University (F2012/IOF-Advanced/
094). K.R. is supported by a European Research Council Starter
Grant ELECTROTALK. We acknowledge Tim Lacoere for the
graphical abstract and Frederiek-Maarten Kerckhof for
developing the R-script and statistical analysis.

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