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Electricity generation from swine wastewater in

microbial fuel cell: Hydraulic reaction time effect

Dongmei Ma a, Zong-Hua Jiang b, Chyi-How Lay b,*, Dandan Zhou c

a
Key Lab of Groundwater Resources and Environment, Ministry of Education, Jilin University, Changchun 130021,
PR China
b
Green Energy Development Center, Master Program of Green Energy Science and Technology, General Education
Center, Feng Chia University, Taichung, Taiwan
c
School of Environment, Northeast Normal University, Changchun 130024, PR China

article info abstract

Article history: Microbial fuel cells (MFC) have emerged as a sustainable technology that can directly
Received 25 March 2016 produce electricity from the oxidation of organic matter by bacteria. A two-chambered up-
Received in revised form flow MFC with inner loop mode is evaluated for the hydraulic retention time (HRT) effect on
2 August 2016 MFC electricity generation. The MFC is constructed with a carbon cloth electrode and anion
Accepted 4 August 2016 exchange membrane for electricity production. Swine wastewater containing
Available online xxx 3300 ± 300 mg/L of total chemical oxygen demand (TCOD) was used as the substrate for the
anode chamber. Potassium ferricyanide was used as the electron acceptor in the cathode
Keywords: chamber. The reactor was operated at 30
C with a external resistance of 100 U in feed-
Swine wastewater batch mode. The TCOD removal rates at HRT 13, 14, and 20 d were 71, 73 and 83%, while
Up-flow the coulombic efficiencies (CE) were 7.1, 2.4 and 0.3%, respectively. The maximum power
Microbial fuel cell density at HRT 13 and 14 d were similar, 12 mW/m2 and 13 mW/m2, which were 26 fold
Hydraulic retention time greater than the HRT at 20 d (0.5 mW/m2). The effect of different HRTs on decreasing the
organic content by biocatalyzing leads to less organic material for electricity generation,
resulting in lower power output. The optimal COD removal efficiency, CE and power
density were observed in an up-flow MFC at HRT 14 d. These results demonstrate that up-
flow MFC using an inner loop with swine wastewater as the substrate could provide both
effective organic removal and electricity output.
© 2016 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.

released into the environment to avoid severe environmental

Introduction
The breeding industry increased with economic development,
leading to increased volumes of animal manure wastewater.
Animal manure wastewater contains high-concentrations of
organics, nitrates and phosphorus. This organic wastewater
must be treated to meet discharge regulations prior to being
pollution from high-strength organic wastewater [1e3]. Many
traditional water treatment techniques are available to
remove pollutants from animal manure wastewater. For
example, physical methods (e.g., precipitation, dewatering),
chemical processing (e.g., coagulation, disinfection), biological
processing (e.g., aerobic and anaerobic treatment) and
resource utilization (composting) [4e7]. Physical and chemical

* Corresponding author.
E-mail address: kfc0612@gmail.com (C.-H. Lay).
http://dx.doi.org/10.1016/j.ijhydene.2016.08.019
0360-3199/© 2016 Hydrogen Energy Publications LLC. Published by Elsevier Ltd. All rights reserved.

Please cite this article in press as: Ma D, et al., Electricity generation from swine wastewater in microbial fuel cell: Hydraulic reaction
time effect, International Journal of Hydrogen Energy (2016), http://dx.doi.org/10.1016/j.ijhydene.2016.08.019
2 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y x x x ( 2 0 1 6 ) 1 e7
processes entail high operational costs, and aeration treat-
ment produces lots of sludge for disposal. Although com-
posting would accomplish resource recycling, the practical
utilization of compost products is limited. Anaerobic treat-
ment would process bioenergy (e.g., CH4 and H2) without
sludge production [6], while its operating conditions are
difficult to control [2]. In fact, breeding industry wastewater is
a promising source of bioenergy with its high level of organics.
Microbial fuel cells are energy-producing devices in which
chemical organics are converted directly into electrical energy
by the catalytic activity of electroactive bacteria [8]. Electro-
active bacteria biodegrade organics and the generated elec-
trons are transported to the anode surface using several
extracellular electron transfer mechanisms, including direct
electron transfer via direct electron transfer via the
membrane-bound c-type cytochrome and/or the bacterial
nanowires, and indirect electron self-transfer mediated by
shuttle molecules [9e11]. The generated electrons are trans-
ferred using an external circuit and accepted by an electron
acceptor in the cathode. The two-chambers (anode and
cathode) are separated by an exchange membrane (e.g., AEM,
CEM and PEM) to allow special ions through the membrane
connected to a closed circuit. The single-chamber MFC has
only an anode while the cathode is exposed to the air, with O2
as the electron acceptor. Recent studies have demonstrated
that the two-chamber MFC has higher mass transfer resis-
tance for the exchange membrane than the single-chamber
MFC without membrane. The two chamber MFC power
output is therefore less than that of the single-chamber under
the same operating conditions [12]. The up-flow MFC is the
most promising sustainable technology having the advan-
tages of retaining very high cell density and high mass
transfer efficiency [13].
The MFC substrate is crucial because it serves as the
nutrient and energy source. The efficiency of converting
chemical energy into bioenergy depends mostly on the sub-
strate composition and the concentration, which influence
the coulomb efficiency and power yields. The major sub-
strates that have been explored in MFCs include various kinds
of artificial and real wastewaters [14]. Real wastewater is the
most meaningful practical implication for the removal of
contaminants with simultaneous electricity generation [15].
This study operated an up-flow two-chamber MFC that
used real pig manure and a swine wastewater mixture as the
anode substrate. The influences of different HRTs on MFC
power yields, COD removal rate and CE are discussed. The
decreased HRT would bring about lower COD removal rate
with increasing max power density and CE. These results
demonstrate the up-flow MFC can degrade swine wastewater
as the energy source with efficient wastewater treatment re-
sults and electricity output.
Materials and methods
Swine wastewater
The pig manure and swine wastewater were collected from a
pig farm located in Central Taiwan and stored separately in a
refrigerator at 4
C prior to use for further experiments. The
Please cite this article in press as: Ma D, et al., Electricity generation from swine wastewater in microbial fuel cell: Hydraulic reaction
time effect, International Journal of Hydrogen Energy (2016), http://dx.doi.org/10.1016/j.ijhydene.2016.08.019
manure was diluted to 10 g/L, and the two solutions were then
mixed at a volume ratio of 1:1 and stirred for 10 min before
filtering using iron mesh (d ¼ mm?) to remove the particulate
organic matter. The filtrate was then used as the anode sub-
strate. The swine wastewater characteristics are shown in
Table 1.
MFC construction and operation
All experiments were conducted using MFCs operated in a
temperature-controlled room (25
C). Fig. 1 shows a schematic
diagram of the two-chamber MFC used in this study. The MFC
reactor was separated into anode (800 mL) and cathode
(300 mL) chambers by an anion exchange membrane (AMI-
7000, Membranes International, Ringwood, NJ). and the anion
exchange membrane contained two sheets of carbon cloth,
6 cm  5 cm and 3 cm  5 cm respectively. A fixed external
resistance (R) of 100 U was connected between the electrodes.
The anode chamber (total length ¼ 21.5 cm, inner
diameter ¼ 6 cm) was downside and operated in inner loop
mode using a circulating pump at a rate of 20 L/h, with a
stirring bar at the bottom to completely mix the electrolyte.
The cathode (total length ¼ 10.7 cm, inner diameter ¼ 6 cm)
was upside with potassium ferricyanide (50 mM K3Fe(CN)6) in
a phosphate buffer (pH ¼ 7.0) used as the catholyte. The
electrolytes for both the anode and cathode were refilled when
the voltage reached lower than 20 mV. The fresh electrolyte
was supplied using a peristaltic pump at the bottom into
anodic and cathodic chambers. The effluent flowed out
simultaneously from the top of each chamber.
Process monitoring
Before and after experiment 1.5 mL samples were siphoned
and centrifuged (1000 rpm, 5 min) for COD measurement
using the closed reflux method.
The potential between the anode and cathode was recor-
ded on a personal computer through a data acquisition system
(JAS-5000, Jiehan Technology Co. Ltd) at 5 min intervals. The
current was calculated from the measured voltage according
to Ohm's law [potential (V) ¼ current (A)  resistance (R)]. The
current was converted into coulombs (C) using this equation
[current (V) ¼ coulomb (C)/time (s)]. The Coulombic efficiency
(CE) (%) is defined as the ratio of total coulombs transferred to
the anode from the anode substrate to the maximum possible
Table 1 e Characteristics of pig manure solution and
swine wastewater.
Item Swine Pig manure solutiona
wastewater
Total COD (g/L) 6e7 8.4e11
NH3-N (g/L) 1e1.3 0.1e0.2
pH 7.1e7.4 6.5e6.8
Conductivity (mS/ 5590 301
cm)
a
The pig manure solution was mixed with 50% of pig manure and
50% of pure water.
 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y x x x ( 2 0 1 6 ) 1 e7 3

Fig. 1 e Schematic diagram of up-flow MFC with inner loop mode reactor.

coulombs present in the removed substrate [16], calculated b is the number of electron moles produced per mol of COD
using (Eq. (1)): (4 mol of e/mol of COD), DS is the COD removal concentration
Pn (g/L), V is the liquid volume (L), M is the molecular weight of
i¼1 Ui ti oxygen (32 mg/L) [8].
CE ¼ M  100% (1)
RFbDSV
Polarization curves were obtained by varying the external
Ui is the MFC output voltage at time ti, R is the external resistances from 1 MU to 5 U using a resistance substitution
resistance, F is the Faraday constant (96,485 C/mol electrons), box. Each resistor was tested for 5 min. The power density was

Fig. 2 e Effect of HRT on Voltage (a: HRT-13; b: HRT-14; c: HRT-20).

Please cite this article in press as: Ma D, et al., Electricity generation from swine wastewater in microbial fuel cell: Hydraulic reaction
time effect, International Journal of Hydrogen Energy (2016), http://dx.doi.org/10.1016/j.ijhydene.2016.08.019
4 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y x x x ( 2 0 1 6 ) 1 e7

Fig. 3 e Effect of HRT on Power output (a: HRT-13; b: HRT-14; c: HRT-20).

calculated according to the following equation: Power density
(mW/m2) ¼ current (mA)  voltage (V)/A (m2), where A is the
surface area of the anode electrode (m2) [9].
Results and discussion
The HRT effect on power generation
A multitude of organics are present in swine wastewater.
Logan demonstrated that swine wastewater can be used for
power generation [10]. The voltage output of different HRT
operating conditions was studied (Fig. 2). In three HRT ex-
periments, when wastewater was added into the MFC, the
voltage output increased immediately. The HRT 13 d (Fig. 2a)
voltage increased from 20 mV to about 90 mV. The maximum
voltage was 92.8 mV during operation. The time period was
almost 2 days. The voltage output trend at HRT 14 d (Fig. 2b)
was similar to HRT 13 d, while the voltage was from 20 mV to
about 59 mV, with a maximum power output maximum of
59.1 mV. The whole time was 1.4 days for each period. How-
ever, for HRT 20 d (Fig. 2c), the voltage output did not show an
obvious change (only from 7 mV to 11 mV), with 1 day for each
period. The voltage output and time period decreased with
HRT from 13 to 20 d. These results demonstrate that HRT
affected power generation and period time for an up-flow
MFC. The lower HRT with more organics maintained in the
system for use by electroactive bacteria produced longer
electricity generation period. Conversely, longer HRT could
only maintain less organic material in the system for elec-
tricity generation during each period. The voltage and time
period were consequently less compared with lower HRT.
While the power output was not consistent with increasing
organic content of in the MFC, lower HRT means greater anode
solution volume had been replaced, more anolyte without
pretreatment added into the reactor, which would bring more
oxygen into the anode. Oxygen serves as the electron acceptor
in the anode and decreases the quantity of electrons that
would transfer to the cathode, causing the power yield to
consequently decrease.
The effect of HRT on power density
The maximum MFC power density and internal resistance are
obtained from the polarization curves. The power density was
normalized to the anode electrode surface area. Fig. 3b and c
shows that the maximum power densities for HRT 13 d and
HRT 14 d were similar, which were 12 mW/m2 and 13 mW/m2,
respectively. However, the HRT-20 power density was lowest,
which was only 0.5 mW/m2 (Fig. 3c). In comparison with these

Please cite this article in press as: Ma D, et al., Electricity generation from swine wastewater in microbial fuel cell: Hydraulic reaction
time effect, International Journal of Hydrogen Energy (2016), http://dx.doi.org/10.1016/j.ijhydene.2016.08.019
 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y x x x ( 2 0 1 6 ) 1 e7 5

Fig. 4 e Effect of HRT on COD removal and CE (a: HRT-13; b: HRT-14; c: HRT-20).

experiments, the open circuit voltage decreased with
increasing HRT. The internal resistance, Rint, was the external
resistance value when the highest power density was reached,
so, the Rint of HRT 13 d, 14 d and 20 d were 2500 U, 900 U and
1000 U, respectively. The power density was affected by the
organic concentration in the MFC reactor. The maximum
power density of HRT 13 d and 14 d were about 24 times that of
HRT 20 d, which was consistent with the voltage trend
because less organic remained in the system for electricity
generation. At for HRT 13 d and HRT 14 d more organics
remained at HRT 13 d than at HRT 14 d, while the internal
resistance was two-fold to HRT 14 d, leading to similar power
density between these two experiments. When sufficient or-
ganics were kept in the reactors, microbes would biodegrade
more organic material for utilization. In this situation more
bacteria would accumulate on the anode surface and thicker
biofilm would conduct, increasing the mass transfer resis-
tance. The internal resistance of the whole system, which
could lower the power generation, would then increase. At for
HRT 20 d the power density was limited by the insufficient
substrate.
The HRT effect on CE and COD removal
The different HRT operating conditions would influence the
CE and COD removal rate (Fig. 4). The HRT 13 d would remove
COD in the range of 59e71%, with the relevant CE at 4.4e7.1%
(Fig. 4b). At HRT 14 d (Fig. 4c) the COD removal rate was
60e73%, and the CE was 1.8e2.4%. At HRT 20 d the COD
removal rate was 71e83%, while the CE was only 0.2e0.3%
(Fig. 4c). These results demonstrate that the COD removal rate
would increase with higher HRT; however, the CE would
decrease. The reasons were: with higher HRT operation, mi-
crobes would biodegrade the limited organics for utilization,
which would improve the COD removal rate. There were two
kinds of microbes in the MFC anode chamber, suspended and
adherent bacteria. The suspended bacteria would biodegrade
organics, while the accumulated microbes, which generated
electricity, would generate electrons via transforming or-
ganics into electricity. The suspended bacteria would utilize
the biodegradable organics at greater rate than the accumu-
lated microbes under insufficient dissolved organic condi-
tions. At HRT 13 d and HRT 14 d, their periods were almost two
days. More dissolved organics were available for electricity
generation, that is, higher CE for them. However, at HRT 20 d
fresh substrate was added within one day, so the CE would
decrease in comparison. In fact, the MFC anode was purged
with Ar for 5 min for the first time utilization, which would
keep dissolved oxygen in the MFC reactor during operation
and weaken the electron transportation to the cathode. As a
result the CE was lower compared with other studies.
Organic matter can be depleted by many reactions in
complicated swine wastewater. The major role was used by
electroactive microorganisms for power generation. The

Please cite this article in press as: Ma D, et al., Electricity generation from swine wastewater in microbial fuel cell: Hydraulic reaction
time effect, International Journal of Hydrogen Energy (2016), http://dx.doi.org/10.1016/j.ijhydene.2016.08.019
6 i n t e r n a t i o n a l j o u r n a l o f h y d r o g e n e n e r g y x x x ( 2 0 1 6 ) 1 e7
Fig. 5 e Electron balance calculation (A: HRT-13; B: HRT-14;
C: HRT-20. P: electron for power generation; R: electron
were consumed by reactor; O: electron of outlet).
substrate can also be removed by alternate electron acceptors,
such as sulfate reduction reaction, heterotrophic denitrifica-
tion and methanogenesis. In fact, the detailed degradation
pathway of organic matter in this system is under testing,
such as methane, NH4 þ , and sulfate analysis.
The electron balance calculation
To calculate the electron balance in the MFC system we used
the inlet COD at different periods. The total electrons were 100
then the outlet electrons were calculated using the remaining
COD. The electrons that had been used were divided into two
parts: (a) transformed into power generation by microorgan-
isms; (b) consumed by the microbial fuel cell reactor and the
ratio of two parts were determined by the CE. These results are
shown in Fig. 5. The organics that were utilized for power
generation were less than the total COD, although more than
60% of the COD was removed in three cells. This ineffective
transformation rate from organics to power was influenced by
the DO contained in the reactors. A higher HRT would lead to
more DO concentration. As a result the oxygen would highly
restrain efficient electron transformation in the anode
compartment due to its competitive impact on the electron
acceptor in the cathode chamber.
These results revealed lower power generation and CE
compared with other researches. This is attributed to the
following factors: (a) Oxygen diffusion in the anode resulted in
substrate loss. (b) Alternate electron acceptors such as sulfate
and nitrate, pert of swine wastewater composition, were
available to decrease the power yield and CE. (c) The produc-
tion of electroactive bacteria was less than that of anaerobic
Table 2 e Comparison of MFC configuration, max power
density (Pmax) and coulombic efficiency (CE) from swine
wastewater.
MFC reactor Pmax (mW/m2) CE (%) Reference
Single chamber 228 e [17]
Single chamber 261 8 [10]
Double chamber 13 7 This study
Please cite this article in press as: Ma D, et al., Electricity generation from swine wastewater in microbial fuel cell: Hydraulic reaction
time effect, International Journal of Hydrogen Energy (2016), http://dx.doi.org/10.1016/j.ijhydene.2016.08.019
bacteria, causing less substrate consumption by electroactive
bacteria for electricity generation [7].
Comparison of different MFC configurations
Comparison of different MFC reactors showed lower power
generation in this research, while the CE was same level as in
other researches (Table 2). The poor power generation could
be caused by the following reasons: (a) the distance between
the two electrodes was approximately 13 cm, which was too
long compared with other MFCs and not beneficial for power
generation [18,19]; (b) the dissolved oxygen in the substrate
would consume part of the electrons when no N2 was purged
into the MFC inlet. Higher power generation would be ach-
ieved by closer electrode distance and less dissolved oxygen in
the reactor.
Conclusions
Un-pretreated swine waste could be used as substrate in an
up-flow MFC. HRT has an effect on power generation and
substrate utilization for the up-flow MFC reactor. Lower HRT
would maintain a large amount of organic matter in the sys-
tem, which would be utilized by bacteria for power yield with
superior voltage value, power density and CE during opera-
tion. The CE value was not as advantageous as in other
studies. CE value improvement would be needed for more
effective power generation and substrate utilization.
Acknowledgements
The authors gratefully acknowledge the financial support by
Taiwan's Ministry of Science and Technology (Grant no. MOST
104-2221-E-035-007).
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Please cite this article in press as: Ma D, et al., Electricity generation from swine wastewater in microbial fuel cell: Hydraulic reaction
time effect, International Journal of Hydrogen Energy (2016), http://dx.doi.org/10.1016/j.ijhydene.2016.08.019