Applied Energy 101 (2013) 213217

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Applied Energy
journal homepage: www.elsevier.com/locate/apenergy

Short-term effects of temperature and COD in a microbial fuel cell

A. Gonzalez del Campo a, J. Lobato b, P. Caizares b, M.A. Rodrigo b, F.J. Fernandez Morales a,
a
b

University of Castilla-La Mancha, Chemical Engineering Department, Edicio Enrique Costa Novella, Avenida Camilo Jos Cela S/N, 13071 Ciudad Real, Spain
University of Castilla-La Mancha, Chemical Engineering Department, ITQUIMA, Avenida Camilo Jos Cela S/N, 13071 Ciudad Real, Spain

a r t i c l e

i n f o

Article history:
Received 9 December 2011
Received in revised form 2 February 2012
Accepted 20 February 2012
Available online 23 March 2012
Keywords:
Microbial fuel cell
Stress test
Temperature
COD
Electricity

a b s t r a c t
In this work, chemical oxygen demand (COD) and temperature stress-tests on a microbial fuel cell (MFC)
were studied. Regarding the temperature stress-test, its value was cyclically modied between 20 and
40 C with stepwise increments of 5 C. The main result was an exponentially increase in the current
intensity generated. In these tests, no hysteresis was observed, indicating that the temperature stress-test
did not modify the behaviour of the MFC used in this work. To study the response of the system under
COD stress conditions, the inuent COD concentration was stepwise modied from the steady-state
value, 100 mg COD L1, to 3000 mg COD L1 and later was reduced stepwise again to 100 mg COD L1.
In these test, it was observed that the higher the COD concentration, the higher the intensity generated.
The electricity yield was an almost constant value of 6.7  106 A mg1 COD removed per hour. In these
tests, hysteresis was observed for the reverse scan, and a hysteresis loop was traced. To study how long
the hysteresis lasts, several stress-tests were carried out during one week, and it was observed that the
hysteresis was maintained for only 2 days. After that, the system recovered the initial behaviour.
2012 Elsevier Ltd. All rights reserved.

1. Introduction
The world population has experienced continuous growth during the last century. Current projections show that with a continuous increase, the population will reach approximately 9 billion
inhabitants by the year 2050 [1]. This population will need to be
supplied with two basic commodities: energy and water. At the
same time, the huge amount of wastes generated by the population
will need to be adequately managed prior to disposal.
During the last century, fossil fuels have been the main energy
source, but its scarcity and high consumption rate as well as the
environmental problems related to the CO2 emissions caused by
fossil fuels have prompted a search for alternative energy sources.
In parallel, the overall reduction in water-source quality suggests
the need for novel technologies for the treatment and reuse of
the wastewaters generated [2]. In this context, renewable energy
sources appear to be one of the ways to reach the proposed energy
objective. Moreover, the use of biomass or wastes as energy
sources seems to be a very interesting option [36]. The organic
substrates contained in the wastes represent a signicant share
of the wastes generated daily [7]; however, its energy cannot be
recovered by traditional processes because of its complex composition and because, in most of the cases, it is highly diluted [8].
Corresponding author. Address: University of Castilla-La Mancha, ITQUIMA,
Chemical Engineering Department, Avenida Camilo Jos Cela S/N, 13071 Ciudad
Real, Spain. Tel.: +34 926 295300x6350; fax: +34 926 295242.
E-mail address: FcoJesus.FMorales@uclm.es (F.J. Fernandez Morales).
0306-2619/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.apenergy.2012.02.064

A promising option to convert the energy contained in the

chemical bonds of the organic wastes to electrical energy is
through the application of Microbial Fuel Cells (MFCs) [9]. MFCs
not only facilitate energy recovery but also remove pollutants at
the same time. Thus, MFCs can also be considered as an environmental technology for waste treatment. MFCs are bioelectrochemical devices in which the microorganisms that are present in the
anodic chamber oxidise substrates, generating electrons, protons,
and other metabolic products. The electrons produced by the
microorganisms are transferred to the anode by electron shuttles,
direct membrane-associated electron transfer, nanowires, etc.
Then, these electrons ow to the cathode through a conductive
material [1012]. In most of the MFCs, the electrons that reach
the cathode combine with protons, which diffuse from the anodic
compartment through the membrane, and with oxygen to produce
water. It is not essential to place the cathode in water or in a separate chamber when using oxygen at the cathode [13] because the
cathode can be congured for direct contact with air [14].
As stated above, oxygen is very often used as the terminal electron acceptor; however, its presence in the anode inhibits electricity generation because microorganisms would oxidise the organic
compounds with oxygen as the nal electron acceptor, avoiding
the external electric circuit, which results in a lower Coulombic
efciency. Thus, the MFC must be designed to keep the bacteria
separated from the oxygen. This separation can be achieved by
placing a membrane that allows proton conduction from the anode
to the cathode, forming two separate chambers: the anodic one
and the cathodic one [15].

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A. Gonzalez del Campo et al. / Applied Energy 101 (2013) 213217

In the literature, experiences with MFCs operating with pure or

mixed cultures are described. It has been reported that MFCs operated using mixed cultures achieved substantially greater power
densities than those with pure cultures [16,17]. Mixed cultures
employ an undened consortium of organisms that grow based
on ecological selection principles, this means that only the tted
organisms will prevail. This approach is very interesting, not only
because of the greater power but also because it reduces the operating costs and makes it possible to deal with wastes [18].
Bacterial reactions can be carried out over different temperature ranges, depending on the tolerance of the bacteria, ranging
from moderate temperatures (1535 C) to both high ranges (40
60 C) and low temperatures (<15 C) and over several COD loads
[10]. Despite the importance of temperature and COD to microbial
metabolisms, there is a lack of information available about them in
the literature related to MFCs. The temperature and COD effects are
of great importance in the practical application of MFCs because
the temperature and COD proles of the wastewaters change in
the short term due to daily changes and in the long term due to
seasonal changes [19].
In this context, the aim of this work was to use a MFC, seeded
with a mixed culture, to study the behaviour of the microorganisms and the performance of the MFC when it was subjected to
temperature and COD stress-test.

The anodic compartment was seeded with activated sludge

from the Ciudad Real Wastewater Treatment Plant [21] and operated in the fed-batch mode. The anodic compartment of the MFC
was connected to a wastewater reservoir of 250 cm3. A peristaltic
pump was used to recirculate the wastewater from the reservoir
through the anodic chamber of the MFC at 2.0 cm3/min. Every
day, 50.0 cm3 of liquid was discarded from the reservoir and replaced by fresh wastewater. The composition of the synthetic
wastewater used in the experiments can be found elsewhere [22].
An air pump was used to supply the required oxygen to the
cathodic compartment. During the experiments, the air pump supplied 6 dm3 min1 of air at 1.5 atmospheres of pressure to the
cathodic chamber.
During normal operation, the anode and the cathode were connected by means of wires and a resistance (125 X). The potentials
between the edges of this resistance were continuously monitored.
These potentials are directly related to the current owing between the electrodes by Ohms law.

IMFC

E125X
125

The COD removal rate (rCOD) in the MFC can be calculated

according to Eq. (2), where V is the volume of liquid of the reservoir
(including the volume of the anodic chamber) and DCOD is the
amount of COD removed in a dened period of time.

rCOD V  DCOD
2. Experimental procedure
2.1. Experimental set-up
The experimental set-up consisted of two chambers, separated
by a Sterion membrane, forming a MFC. The anodic and cathodic
electrodes were based on Toray carbon papers TGPH-120 (E-TEK,
USA), the anodic one with 20% Teon content and the cathodic
one with 10%. In the cathode, a catalytic layer with 0.5 mg Pt/cm2
loading was deposited onto a microporous layer because of the
advantages that this additional layer offers; this material has been
described in the literature [20]. Both the anodic and cathodic
chambers were built on a graphite plate. The volume of the anodic
chamber was 0.95 cm3, and its active area was 4.65 cm2. The volume of the anodic chamber was 0.50 cm3, and its active area was
2.85 cm2. The membrane-electrode assembly was formed by hot
pressing. The membrane and the electrode were introduced between two stainless steel blocks equipped with heating surfaces
and a temperature control system. Hot-pressing was performed
at 130 C by applying a load of 1 ton for 15 min. A schematic view
of the set-up is shown in Fig. 1.

Taking into account the rCOD and the Faraday constant (96,485
C mol1 e), the molecular weight of oxygen (M) and the stoichiometry (n) of the oxidation of the chemical oxygen demand (every
mmol of COD corresponds to 4 mmol of e), Eq. (3) can be used to
calculate the maximum current that can be produced from COD oxidation (Istoichiometric).

Istoichiometric mA


1

n4 mmol e  mmol

O2  F96485 mC mmol e
1

1

Mmg O2  mmol O2  86400s  d

1

 rCOD mgO2  d

2.2. Characterisation techniques

The pH, conductivity and dissolved oxygen were measured
using a GLP22 Crison pH-meter, an LF538 WTW conductivitymeter and an Oxi538 WTW oxy-meter, respectively. The total suspended solids and COD were measured according to Rodrigo et al.
[23]. A digital multimeter was connected to the system to

Fig. 1. Schematic view of the set-up.

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A. Gonzalez del Campo et al. / Applied Energy 101 (2013) 213217

continuously monitor the value of the cell potential. Polarisation

curves were recorded using a Voltalab 21 potentiostat/galvanostat
(Radiometer Analytical).
The polarisation curves from the MFC test provide interesting
information about the operating conditions of the MFC, in particular, about the actual capabilities of the MFC. These curves make it
possible to discern three important parameters: the open circuit
voltage (OCV) or the maximum allowable MFC voltage (for a zero
current), the maximum intensity reachable (for a zero potential)
and the maximum feasible power density. In addition, the shape
of the curve gives information about the limiting stage that controls the performance of the cell. Typically, a polarisation curve
can be divided into three zones, but in MFCs, linear polarisation
curves are most often obtained, which suggests that the real mechanisms occurring inside them are a mixture of those occurring in
conventional fuel cells [24,25]. The main source of error in the
characterisation techniques used in this work was due to the variability in the behaviour of the microorganisms of the anodic
biolm.
3. Results and discussion
3.1. Short-term temperature effect
The effect of temperature on the cell voltage and the electrochemical behaviour of the MFC were studied. Experiments were
carried out at different temperatures during a brief period of 5 h;
meanwhile, the rest of the variables were kept constant in both
the anodic and cathodic compartments of the MFC. The operational
temperature was stepwise increased from 20 to 40 C, with step
increments of 5 C, and afterwards, the temperature was stepwise
decreased with step decrements of 5 C.
The results of the experiments are shown in Fig. 2A, in which
the variation of the cell voltage with time due to changes in the
temperature is shown.
In Fig. 2A, it can be observed that the increase in the temperature signicantly increased the cell intensity. All of the results of
the cell voltage generated with each temperature are represented

(Fig. 2B). In this gure, it can be observed that the modication

of the temperature, within the range of temperatures studied in
this work, generated a plot without hysteresis. The absence of hysteresis in this gure means that the system behaved in an identical
manner before and after the temperature stress-test study, indicating that modifying the temperature does not have an effect on the
long-term behaviour.
The increase as a function of temperature shown in Fig. 2 could
be related to the enhancement of the microbial metabolism, to the
enhancement of the membrane permeability and to the Ohmic
resistance reduction due to the higher conductivity of the liquid
solution. To study these effects, the contributions of the membrane
conductivity, the Ohmic resistance and the microbial metabolism
were analysed.
Regarding the Sterion membrane permeability, it was previously observed that a modication in the temperature within the
range studied in this work barely inuenced the permeability
[26]. Therefore, the electricity generation of the MFC should not
be sensitive to the change in the membrane permeability caused
by the increment in the temperature applied in this work.
Regarding the conductivity, the temperature effect on this
parameter also presents a linear trend, with the average increase
in the conductivity for ionic solutions being approximately
2% C1 [27]. Because of the inverse relationship between the conductivity and the resistance, and considering that the internal
resistance of the MFC is not caused by only the electrolyte, a linear
relationship with a decrease in the internal resistance of the MFC
with a value up to 2% C1 was expected. This relationship was
conrmed by the analysis of the polarisation curves (see Fig. 3)
and by the calculations derived from these curves (see Table 1).
In this work, the internal resistance of the MFC decreased by
approximately 0.6% C1. This decrease could be explained by the
following observation: the higher the temperature, the higher the
ionic conductivity, and therefore, the lower the Ohmic resistance
of the MFC. However, the change in the Ohmic resistance is not sufcient to explain the changes in the intensity with temperature; in
addition, these changes are not linear but exponential.
It is known that the temperature effect on microorganism activity yields an exponential trend. In this sense, the exponential trend
observed in this experiment indicated that most of the effect

Fig. 3. MFC performance at different temperatures.

Table 1
Effect of temperature over the MFC performance.

Fig. 2. (A) Dependence of cell intensity on anodic cell temperature. (B) Cyclical
relation of cell intensity with anodic cell temperature.

T 1 (C)

OCV (mV)

Pmax (mW/m2)

IPmax  106 (A)

Rint (KX)

20
25
30
35
40

111
112
117
119
133

0.73
0.75
0.82
0.88
1.01

8.49
8.56
8.96
9.65
10.1

15.36
15.04
14.55
14.16
13.45

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A. Gonzalez del Campo et al. / Applied Energy 101 (2013) 213217

observed over the intensity generated by the MFC was caused by

the enhancement of the microbial activity, which supports the
statements related to the membrane permeability and Ohmic
resistance exposed above.
It is well known that the relationship between the reaction rate
constant and temperature can be expressed by the Arrhenius equation, where A is the pre-exponential factor, Ea (J/mol) is the activation energy of the reaction and T (K) is the absolute temperature.
Taking into account that the intensity is proportional to the reaction rate, the Arrhenius equation can also be used to determine
the temperature effects on the intensity generated by the MFC.
Ea

I A  eRT

Taking the natural logarithm of the Arrhenius equation yields the

following:

LnI LnA 

Ea 1

R T

Thus, when the microbial reaction has a constant rate, and therefore
a constant intensity that obeys the Arrhenius equation, a plot of
Ln(I) versus T1 should give a straight line, whose slope and intercept can be used to determine Ea and A. This representation was
made by obtaining an Ea of 1.11  104 J/mol and a pre-exponential
factor of 1.96  101 mA; the R2 value for the regression was 0.96.
Unfortunately, it was impossible to compare these values with
those obtained in other works carried out with microbial fuel cell
cultures because there is no information in the scientic literature
regarding the microorganisms present in MFC.
The sensibility of the intensity to increasing temperature is very
important to model biological processes and can be determined
using modications of Eq. (4). An example is Eq. (6), where h is
the temperature constant.
 C

IT  C I20  C  hT20

Under the studied conditions, the h value was 1.011 (C1), a value
very close to the typical value when working with mixed cultures
based on heterotrophic microorganisms [14]. Another important
parameter is the temperature coefcient, QT, which corresponds
to the ratio of the intensity generated at different temperatures in
a selected range, usually 10 C.

Q 10

IT 10  C
IT  C

3.2. COD effect

The effect of the COD concentration on the liquid bulk of the
anodic compartment over the intensity for the maximum power
generation and on the electrochemical behaviour of the system
was also studied. Experiments were carried out at different COD
concentrations, during 5 h periods, while the rest of the variables
in both the anodic and cathodic compartments of the MFC were
held constant. The lengths of these periods were so low to avoid
long-term modications to the system, mainly excessive biomass
growth. The COD concentrations tested in these experiments were
stepwise increased from 100 mg COD/dm3, which was the COD
concentration maintained during the steady-state operation of
the cell, to 3000 mg COD/dm3. Afterwards, the COD concentration
was decreased stepwise, following the same pattern, from
3000 mg COD/dm3 to 100 mg COD/dm3.
The experimental results obtained are presented in Fig. 4A.
In this gure, it can be observed that the higher the COD, the
higher the intensity generated. However, it is important to note
that the behaviour of the cell changed as the experiment progressed, generating a hysteresis loop (see Fig. 4B).
In short-term experiments, the hysteresis loop could be explained by the synthesis of enzymes during the high load periods
or by the enhancement of the ability of the microorganisms to produce electricity. The latter statement was rejected because it was
observed in all tests that the electricity-yield presented an almost
constant value of 6.7  106 A mg1 COD removed per hour. Therefore, the most probable explanation for the hysteresis loop is that
during these tests, the microorganisms increase their enzymatic
production to degrade as much COD as possible. However, even
when the high COD concentration had disappeared the ability
developed by the microorganisms to synthesise more enzymes
was, for some time, maintained. Therefore, in these situations,
the microorganisms were able to degrade the COD at high rates,
increasing the maximum intensity produced by the MFC.
With the aim to understand this effect, the length of this
enhancement in the enzyme synthesis was studied. COD stresstests were repeated with a break between tests of 2 and 4 days.
The experimental results obtained are presented in Fig. 5.

This coefcient determines the change in the cell voltage along with
the change in temperature. In this work, the calculated temperature
coefcient was 1.12, indicating that an increase in the temperature
of 10 C caused a 12% increase in the intensity generated by the
MFC.
In these experiments, it was also observed that with the increase in the temperature of the anodic compartment of the
MFC, the OCV, the maximum power and the intensity for the maximum power generation increased (see Table 1). This result could
be explained because the higher the temperature, the higher the
microbial metabolism and therefore the higher the electrical current and maximum power. The main utility of these ndings related to the temperature effects is the fact that MFC can be
operated under different temperature values being accurately predictable the electrochemical behaviour of the system. This means
that the MFC is a very robust device when it is subjected to
short-term changes in the operational temperature. Moreover, in
the case of the use of these electrochemical devices as sensors,
these sensors would be very robust and could be used even for
medical applications.

Fig. 4. (A) Dependence of maximum intensity on COD concentration. (B) Hysteresis

curve for a COD stress-test.

A. Gonzalez del Campo et al. / Applied Energy 101 (2013) 213217

217

References

Fig. 5. Long term dependence of maximum intensity on COD concentration.

In this gure, it can be observed that 2 days after a COD stress

test, the microorganisms still presented the ability to degrade the
COD at a higher rate, therefore generating a higher intensity. However, 4 days after the stress-test, the microorganisms generated
lower intensities for the same operating conditions. This result
indicated that, after four days, the microorganisms returned to
the enzymatic levels that were maintained before the COD
stress-test. The importance of this nding is that the electrochemical behaviour of the system (exerted current, OCV, power density,
etc.) is highly affected by the previous COD loads, hindering the
accuracy in the electricity generation prognosis. In the use of
MFC as sensors, the dependence on the previous COD concentrations experienced must be taken into account because it would
cause perturbations in the measurement.
4. Conclusions
From the results obtained in the present work, the following
conclusions can be made:
The modication in the operation temperature of the anodic
compartment of an MFC modies the response, causing an
increase in the temperature and, within the studys range, an
increase in the intensity. This response is mainly due to the
higher microbial activity, although the higher conductivity of
the membrane also has an effect. Regarding the temperature
effects, is important to remark that the temperature modications did not modify the system in the long term.
The modication in the COD concentration of the inuent fed to
the anodic compartment also caused modications in the
behaviour of the cell. The increase in the COD concentration
enhanced the activity of the microorganisms, which increased
the current of the cell. This enhancement in the activity of the
microorganisms was maintained for at least 2 days. However,
its effect disappeared after 4 days, indicating that the microorganisms returned to the previous enzymatic levels.

Acknowledgement
The authors thank the JCCM for the nancial support through
Project POII10-0329-5194.

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