Bartolome 2023 IOP Conf. Ser. Earth Environ. Sci. 1146 012004

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4th International Conference on Resources and Environment Sciences (ICRES 2022) IOP Publishing
IOP Conf. Series: Earth and Environmental Science 1146 (2023) 012004 doi:10.1088/1755-1315/1146/1/012004
Optimization of bioenergy generation and chemical oxygen

demand reduction in dual-chamber microbial fuel cells using
active soil microbes
G J C Bartolome1 and L L Tayo2
1 Department of Engineering, Cavite State University-Cavite College of Arts and

Trades, Tejeros Convention, Rosario, Cavite, Philippines 4106
2 Biological Engineering, School of Chemical, Biological and Materials Engineering
and Sciences, Mapúa University, Muralla St., Intramuros, Manila, Philippines 1002
E-mail: lltayo@mapua.edu.ph
Abstract. This study focused on the optimization of a dual-chamber microbial fuel cell (MFC)
system using carbon fiber brush and titanium electrodes, meat processing wastewater as
substrate, and active soil microbes as biocatalyst. Significant findings revealed promising results
on the MFC performance. The active soil microbes contain viable bacteria that worked
effectively in the anode chambers. Peak open-circuit voltages (OCV) of the MFCs were as high
as 1.05 V using titanium mesh and 1.03 V using carbon fiber brush. In closed-circuit conditions,
the current obtained were 1.04 mA and 1.38 mA, and the power densities were 1.22 Wm -2 and
0.34 Wm-2, correspondingly. The power densities were normalized based on the surface areas of
the anodes. The bioenergy generation profiles also revealed the MFCs can produce peak
potentials of 30.109 kJ and 19.415 kJ using carbon fiber brush and titanium mesh, respectively.
Furthermore, the results showed that the reduction in chemical oxygen demand (COD) was
highest at 24.32 percent using carbon brush and lowest at 13.34 percent using titanium mesh
electrodes.
1. Introduction
The presence of exoelectrogenic microorganisms in microbial fuel cells allows the process of
decomposition of different substrates and compounds and among the by-products of the electrochemical
processes is electricity. Through microbial action, the chemical energy content of the wastewater is
converted into electrical energy while treating the wastewater treatment to improve its quality. Under
mild conditions, MFCs can work efficiently without the need for expensive catalysts. There has been a
variety of MFC configurations available, but the dual-chamber arrangement is more advantageous in
observing the disintegration of compounds during experiments.
A classical dual-chamber MFC has an anaerobic anodic chamber where the microorganisms oxidize the
substrate generating proton and electron equivalents (1) and the anaerobic cathodic chamber where the
reduction reaction happens and produces water as a by-product (2-3). The compartments are linked by
either a proton exchange membrane, cation exchange membrane (CEM), or a salt bridge which allows
protons to move across to the cathode chamber while preventing the flow of oxygen to the anode.
The performance of MFC is influenced by several factors among which is the electrode materials that
largely impact the cost of the system (4). Platinum electrodes are common and superior to graphite and
carbon but are more expensive. Carbon fiber brush, on the other hand, are more advantageous compared
with other materials such as graphite grains, carbon cloth, and activated carbon felt by generating higher
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power densities. Carbon brushes are also more recommended for scaled-up applications as they are more
porous, have higher surface areas, and can easily be manufactured relative to brush size (5).
In terms of microbial inoculants in the MFCs, mixed cultures are generally preferred for practical
applications because they perform better on power generation (6), they are more accessible in large
quantities, have a higher tolerance to environmental fluctuation, and are more responsive to different
substrates (7). The use of active microorganisms in MFCs does not require complex and expensive
isolations. Under suitable conditions, the microorganisms can grow and maintain their metabolic
activities. Effective microorganisms (EM) are one of the various types of microbial inoculants that can
be found in the market.
As a viable technology for the generation of renewable energy and treatment of wastewater, the interest
in MFCs has grown enormously in recent years, both in terms of the numbers of research and practical
applications (8). Among the applications of bioelectrochemical systems, MFCs have received the most
attention because of their ability to treat wastewater and generate electricity simultaneously without the
need for external chemical amendments and with very little to zero external energy. This study focused
on the investigation of the performance of microbial fuel cells using meat processing wastewater as
substrate inoculated with active soil microbial consortium derived from the activated solution of EM. In
both foreign and local settings, there is a scarcity of published literature that focuses on the use of meat
processing wastewater and active soil microbes (effective microorganisms) in microbial fuel cells.
While EM is locally available, meat establishments are also widespread in the Philippines, which
suggests that there is an abundance of wastewaters from meat processing activities. The study
demonstrated the effectiveness of the active soil microbes in generating bioenergy from meat processing
wastewater. The MFCs were operated using two types of electrode materials ̶ carbon fiber brush and
titanium mesh ̶ with starch as co-metabolite and methylene blue as the electron mediator. The effects of
these components on the MFC performance were described in terms of electrode potential, current,
power density, bioenergy generation, and reduction of chemical oxygen demand (COD) of the
wastewater
2. Methodology
2.1 Construction of the microbial fuel cells (MFCs)

Dual-chambered MFCs made from 250-ml Pyrex reagent bottles were used in the experiment. Each pair
of these reagent bottles that served as the cathode and anode chambers were connected by a flange to
resemble a dual-chamber arrangement while holding the cation exchange membrane (CEM). Both
chambers were modified to introduce an inlet and outlet port on one side where the substrate passes and
top holes on the screw cap for aeration and buffering. Pretreatment and treatment methods followed the
procedure in a published literature (9).
2.2 Pre-treatment of electrodes

The performance of MFCs was compared using two types of electrodes. To remove the impurities,
carbon brushes were soaked in pure acetone overnight. For the acid treatment, brushes were soaked in
a solution of 200 gL-1 (NH4)2S2O8 and 1 M H2SO4 for 15 min. Using a forced convection oven, brushes
were finally heat-treated at 200oC for 30 min. The carbon brushes were washed with deionized water
several times before application in the MFCs. The other electrode is a cylindrical titanium mesh welded
on stainless steel core.
2.3 Preparation of the microbial inoculant

In preparing the inoculant, one liter of EM1 (effective microorganism solution) was activated by
fermentation for 7 days. EM1 was mixed with 30 L of non-chlorinated water and three kilograms of
molasses then transferred to an airtight carboy to ferment. On the 3rd day of fermentation, the carboy
was slightly opened to allow some gases to escape. The inoculant was transferred to 500-mL sterile
bottles for proper storage.
2
2.4 Optical density measurement of the microbial inoculant

The viability of the active soil microbes in the inoculant was investigated by optical density
measurement (OD600) using a 722G single-beam visible spectrophotometer (600 nm wavelength). A
serial dilution of the inoculant was prepared by diluting 10 ml of the activated solution to one liter with
non-chlorinated water. The transmittance of the samples from the dilution series was taken at time zero
and regular intervals of one hour for 10 hours.
2.5 Collection and characterization of meat processing wastewater

Meat processing wastewater (MPW) was taken from the treatment plant of a meat processing and cold
storage facility located in Carmona, Cavite. Fresh, unsettled influent was taken and transported to the
laboratory. In the interest of establishing the operation of the MFC for a reduction in chemical oxygen
demand, wastewater samples were submitted to JefCor Laboratories, Inc for assessment of initial COD.
2.6 Operation of the MFC

The MFC systems were operated in batch mode in ambient temperature and pressure. For the
measurement of open-circuit voltage, MFCs were operated in open circuit conditions. On the other hand,
for the measurement of current, power, and estimation of the bioenergy generation, MFCs were operated
in closed circuit conditions at a constant external load of 200 Ω. CEMs were replaced after each batch
of operation to minimize contamination between anode and cathode chambers. While being aerated
continuously, the cathode was also regularly buffered with 0.01 KMNO4 solution to minimize changes
in pH.
Experiments were conducted in three stages. In stage 1, to determine the effects of different
concentrations of starch (as co-metabolite), the pure microbial inoculant were acclimatized in the anodes
and fed with different concentrations of soft starch from zero to 200 ppm, 400 ppm, 600 ppm, and 800
ppm. In stage 2, to determine the effects of methylene blue as an electron mediator, the MFCs which
recorded the highest power densities based on the concentration of starch were spiked with methylene
blue ranging from zero to 0.5, 1.0, 1.5, and 2.0 mM concentration. In stage 3, the MFCs were switched
to meat processing wastewater (MPW), starch, and methylene blue. The effects of different
concentrations of microbial inoculant on power generation were investigated by feeding the anodes with
different ratios of MPW and microbial inoculant from 1:0, 1:3, and 1:1.
2.7 Data gathering procedure

The operation of the MFCs was monitored in real-time using data logging equipment. The system
records the open-circuit voltage, voltage drop, and current in both open and closed circuit conditions.
The data collected by the equipment was randomly checked with a digital multimeter. The power density
for each MFC was normalized to the estimated cross-sectional area of the anodes. Using the equations
below, power, power density, and energy were calculated:
P = I*V (1)
P
PD = A (2)
E = P*t (3)
where P is power (W), I is current (A), V is electrode potential (V), Pd is power density (W m-2), A is
the surface area of the anode (m2), E is energy transferred (J) and t is fermentation time (s).
Lastly, the reduction in COD was computed from the initial and final COD of the analytes determined
by a third-party testing laboratory.
3. Results and Discussion
3.1 Cell viability of the active soil microbes

The transmittance and the absorbance curves of the samples from the dilution of the microbial inoculant
depict the exponentially increasing cell density of the microorganisms in the dilution. The transmittance
3
significantly decreased from 100 percent during the 1st hour to 30 percent in the 10th hour while
absorbance increased from zero to 0.523 absorbance units (AU) in the same period.
3.2 Open circuit voltage of the MFC systems

The open-circuit voltage (OCV) profiles of the MFC-A (with carbon fiber brush) and MFC-B (titanium
mesh) revealed high OCVs. Within the 24 hours, MFC-A peaked at 824 V from initially 708 mV. MFC-
B on the other hand peaked at 1,140 mV from 996 mV.
The maximum OCV achieved by the MFC systems was compared with previous studies that employ
dual-chamber MFCs with different substrates and inocula (10). Carbohydrate-rich substrates such as
glucose, xylose, and glucuronic acid inoculated with mixed bacterial culture produced 390, 380, and
440 mV, respectively. Polyalcohols like galactitol and ribitol inoculated with mixed bacterial culture
generated 340 and 320 mV. Propionate fed in anaerobic sludge obtained 800 mV while acetic acid fed
in a mixed culture of bacteria obtained 475 mV. Glucose inoculated with Saccharomyces achieved
higher OCV at 887 mV (11). Other compounds such as amino acids, nitrogenous compounds, alcohols,
sulfides, phenols (10), and proteins (12) have also been used in MFCs.
3.3 Performance of MFCs as affected by different concentrations of starch

Under closed-circuit conditions, MFC-A spiked with 600 ppm of starch registered an average current of
0.37 mA with a peak at 0.44 mA. This current output was significantly higher among other setups (p =
0.000). Meanwhile, the average current was recorded at 0.29 mA with 800 ppm starch, followed by 0.27
mA for 400 ppm, 0.20 mA for zero ppm, and 0.15 mA for 200 ppm.
For MFC-B, the highest average current of 0.53 mA was recorded from the setup containing zero ppm
starch which is significantly higher among others (p = 0.000). The peak current was measured at 0.58
mA. For other MFCs, an average current of 0.49 mA was obtained from the 200 ppm, 0.45 mA from
the 600 ppm, and 0.44 mA in the 400 and 800 ppm.
3.4 Performance of the MFC as affected by methylene blue (electron mediator)

The presence of methylene blue in MFC-A at 0.5 mM concentration demonstrated a positive effect on
current output by substantially increasing the current from 0.12 mA to 0.28 mA during the 1 st hour of
the batch operation at a visibly stable energy flow. On contrary, other MFCs exhibited high current
during the start of the MFC operations which decreased over time. Nonetheless, the average current was
significantly higher without an electron mediator (p = 0.000).
Comparatively, in MFC-B current was initially lower when anolytes were spiked with methylene blue
which increased throughout the batch operation. Among all non-mediator-less setups, the MFC with 1.0
mM of methylene blue showed higher current values, particularly at an average of 0.44 mA. The same
with MFC-A, the average current remained significantly higher without an electron mediator (p =
0.000).
Significant findings with mediatorless MFCs were parallel with the majority of published studies that
achieve higher rates of electron transfer even without the presence of electron shuttles. One reason is
that some microbes have been identified to form biofilms on the anode which transfer electrons by direct
conductance or by utilizing naturally occurring compounds in the anode from redox reactions.
3.5 Power and energy generation of mediatorless MFCs

Earlier findings revealed that the performance of the MFCs was better in the absence of an electron
mediator. For MFC-A, the average power density was estimated at 0.027 Wm-2, while for MFC-B, the
average power density was 0.353 Wm-2.
The power density values for MFC-A (with 600 ppm of starch) and MFC-B (with no starch) were
normalized by the surface area of the electrodes which is equal to 1.04 m 2 for carbon brush and 0.162
m2 for titanium mesh. Thus, MFC-B performs better yielding significantly higher power densities. This
performance was also comparable with previous studies (10) (Figure 1). This only shows that the surface
area of the electrodes has a significant effect on power density. A smaller surface will yield higher power
density considering constant voltage drop and electron flow.
4
In the same manner, energy profiles for the abovementioned MFC systems show an increasing trend
throughout their operation. Peak energy values were 3.345 kJ for MFC-A and 5.039 kJ for MFC-B.
Bioenergy potential was substantially higher for the MFC with titanium mesh electrodes.
Figure 1. Comparison of maximum power density values in Wm-2 of MFCs

fed with simple and complex substrates.
3.6 Performance of the MFC using MPW and microbial inoculant
3.6.1 Open circuit voltage. The MFC systems fed with full strength MPW demonstrated high open-
circuit voltage. The average potential difference was significant between the two systems (p=0.003)
wherein MFC-A obtained an average OCV of 922 mV lower than MFC-B with an average of 1,005 mV.
Maximum OCVs were 1,032 mV and 1,052 mV for MFC-A and MFC-B, respectively. However, it was
observed that carbon brushes allowed a more stable voltage generation during operation. These values
were also found comparable with previous studies (27, 29) (Figure 2).
5
Figure 2. Comparison of maximum open-circuit voltage (OCVmax) of MFC systems

fed with different wastewaters as substrates.
3.6.2 Current profile. A significant difference in the average current influenced by varying ratios of
MPW and inoculant was observed in MFC-A (p = 0.000) but not in MFC-B. Results showed that the
ratio of 1:1 yields a higher current. Here, the current peaked ranged between 1.22 mA to 1.38 mA with
an average of 1.326 mA.
3.6.3 Power density and bioenergy generation. Power densities were estimated for MFC-A and MFC-
B containing a 1:1 and 1:3 ratio of MPW to microbial inoculant. In here, MFC-B shows significantly
higher average power density at 1.223 Wm-2 (t (24.164) = (-) 32.610). Comparing the results with
previous studies, the MFCs showed promising performance at par with other systems, especially for
MFC-B.
The corresponding bioenergy generation profiles of the MFC systems demonstrated increasing and high
energy values for both MFCs with an average of 15.609 kJ for MFC-A and 9.440 kJ for MFC-B. Peak
energy values were 30.109 kJ and 19.415 kJ for MFC-A and MFC-B, respectively.
6
Figure 3. Comparison of maximum power densities (Pdmax) obtained from different MFC systems fed
with different substrates and inoculants.
3.7 Chemical oxygen demand (COD) removal

The chemical oxygen demand (COD) in ppm in the anolytes of the MFC systems was compared (Figure
4). The original COD of the wastewater was 3,326 ppm. There was an increase in the initial COD of the
anolytes in other MFC setups (1:3 and 1:1 ratio of MPW to microbial inoculant) attributed to the
increasing concentration of the inoculant. Among the MFCs observed, MFC-A delivered a higher COD
removal at 24.23 percent (1:0). The COD percentage removal was lowest using titanium mesh in the
same condition with only 13.34 percent.
7
Figure 4. Initial and final concentrations of COD in the anolytes of different MFC setups
(in mg/L).
4. Conclusion and Recommendations

The results of the study demonstrated the wide abilities of the active soil microbes from EM inoculants
in generating bioenergy in dual-chamber MFCs. Significant results revealed that we can optimize the
operation of these MFCs using meat processing, starch as co-metabolite, and electrode materials for
bioenergy generation and COD reduction. The MFCs demonstrated high open-circuit potentials
comparable with other systems. Using carbon fiber brushes, the MFCs achieved higher and more stable
current generation, higher bioenergy generation potential, and greater COD reduction from the
wastewater. The titanium mesh electrode was instrumental in achieving higher power density and
bioenergy. Lastly, methylene blue as an electron mediator did not significantly increase the power
generated in the MFC systems.
5. References
[1] Sudarsan, J., Prasana, K., Nithiyanantham, S. and Renganathan, K. (2015). Comparative study of
electricity production and treatment of different wastewater using microbial fuel cell (MFC).
Environment and Earth Science, 73, 2409-2413.
[2] Logan, B., Hamelers, B., Rozendal, R., Schroder, U., Keller, J., Freguia,…,Rabaey, K. (2006).
Microbial fuel cells: Methodology and technology. Environmental Science and Technology,
40(17), 5181-5192.
[3] Higgins, S.R., Lopez, S.J., Pagaling, E., Yan, T., and Cooney, M.J. (2013). Towards a hybrid
anaerobic digester-microbial fuel cell integrated energy recovery system: An overview of the
development of an electrogenic biofilm, Enzyme and Microbial Technology, 52, 344-351.
[4] Zhou, M., Chi, M., luo, J., He, H., and Jin, T. (2011). An overview of electrode materials in microbial
fuel cells. Journal of Power Sources, 196(10), 6029-6035.
8
[5] Logan, B., Cheng, S., Watson, V., and Estadt, G. (2007). Graphite fiber brush anodes for increased
power production in air-cathode microbial fuel cells. Environmental Science and Technology, 41,
3341-3346.
[6] Rabaey, K., Boon, N., Siciliano, S.D., Verhaege, M., and Verstraete, W. (2004). Biofuel cells select
for microbial consortia that self-mediate electron transfer. Applied Environmental Microbiology,
70, 5373-5382.
[7] Ahmed, M., Zhou, J., Ngo, H., Guo, W., Thomaidis, N., and Xu, J. (2017). Progress in the biological
and chemical treatment technologies for emerging contaminant removal from wastewater: A
critical review. Journal of Hazardous Materials, 323, 274-298.
[8] Pant, D., Bogaert, G., Diels, L., and Vanbroekhoven, K. (2009). A review of substrates used in
microbial fuel cells (MFCs) for sustainable energy production. Bioresource Technology, DOI:
doi:10.1016/j.biortech.2009.10.017
[9] Bartolome, G. J. and Tayo, L. L. (2020). Decolorization of methyl orange using a double- chamber
microbial fuel cell with the use of soil microorganisms. IOP Conf. Series: Earth and
Environmental Science 563 (2020) 012007 doi:10.1088/1755-1315/563/1/012007
[10] Pandey, P., Shinde, V., Deopurkar, R., Kale, S., Patil, S., and Pant, D. (2016). Recent advances in
the use of different substrates in microbial fuel cells toward wastewater treatment and
simultaneous energy recovery. Journal of Applied Energy, 168, 706-723.
[11] Permana, D., Rosdianti, D., Ishmayana, S., Rachman, S., Putra, H., Rahayuningwulan, D., and
Hariyadi, H.R. (2015). Preliminary investigation of electricity production using dual chamber
microbial fuel cell (dcmfc) with Saccharomyces cerevisiae as biocatalyst and methylene blue as
an electron mediator. Procedia Chemistry, 17, 36-43.
[12] Logan, B. and Heilmann, J. (2006). Production of electricity from proteins using a microbial fuel
cell. Water Environment Research, 78, 531-537. DOI:10.2175/106143005X73046

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This content was downloaded from IP address 136.158.17.179 on 11/09/2023 at 10:31

Optimization of bioenergy generation and chemical oxygen

G J C Bartolome1 and L L Tayo2

1 Department of Engineering, Cavite State University-Cavite College of Arts and

2.1 Construction of the microbial fuel cells (MFCs)

2.2 Pre-treatment of electrodes

2.3 Preparation of the microbial inoculant

2.4 Optical density measurement of the microbial inoculant

2.5 Collection and characterization of meat processing wastewater

2.6 Operation of the MFC

2.7 Data gathering procedure

3. Results and Discussion

3.1 Cell viability of the active soil microbes

3.2 Open circuit voltage of the MFC systems

3.3 Performance of MFCs as affected by different concentrations of starch

3.4 Performance of the MFC as affected by methylene blue (electron mediator)

3.5 Power and energy generation of mediatorless MFCs

Figure 1. Comparison of maximum power density values in Wm-2 of MFCs

3.6 Performance of the MFC using MPW and microbial inoculant

Figure 2. Comparison of maximum open-circuit voltage (OCVmax) of MFC systems

3.7 Chemical oxygen demand (COD) removal

4. Conclusion and Recommendations

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