Process Safety and Environmental Protection 161 (2022) 357–373

Contents lists available at ScienceDirect

Process Safety and Environmental Protection

journal homepage: www.elsevier.com/locate/psep

Microbial fuel cells: Insight into simultaneous wastewater treatment and

bioelectricity generation ]]
]]]]]]
]]

⁎
Ali Nawaz a, Ikram ul Haq a, Kinza Qaisar a, Burcu Gunes b, , Saleha Ibadat Raja a,
Khola Mohyuddin a, Haseeb Amin a
a
Institute of Industrial Biotechnology, Government College University, Lahore 54000, Pakistan
b
School of Biotechnology and DCU Water Institute, Dublin City University, Dublin, Ireland

a r t i c l e i n fo a bs tr a ct

Article history: Burgeoning industrialization has escalated the energy demand and amplified the insistence on depleting
Received 27 January 2022 petroleum-based reservoirs. Subsequently, this has imposed a study to pinpoint renewable alternative
Received in revised form 13 March 2022 sources. Employing renewable resources promotes energy production coupled with environmental sus­
Accepted 14 March 2022
tainability. Microbial fuel cells (MFCs) have an encouraging role in producing sustainable and viable elec­
Available online 16 March 2022
trical energy. It transforms chemical energy into electrical energy with the assistance of microorganisms
acting as biocatalysts. It is an emerging technology that has the potential to treat wastewater with bioe­
Keywords:
Microbial fuel cell, wastewater treatment lectricity production simultaneously. However, scaling up this technology is a major challenge as a plethora
Bioelectricity of technical constraints limits its application in the real world. This review highlights MFC performance in
Biocatalysts terms of power generation by utilizing various wastewaters as substrate. Moreover, it sheds light on dif­
Electrogens ferent MFC designs, mechanisms, and parameters affecting MFC performance along with their pros and
Reactor design cons to generate maximum power output. It concludes possible ways to combat its drawbacks and discusses
its future prospects.
© 2022 The Author(s). Published by Elsevier Ltd on behalf of Institution of Chemical Engineers.
CC_BY_4.0

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 358
2. What are microbial fuel cells (MFCs)? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359
3. Bioelectricity generation from wastewater via MFC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359
4. Electron transfer mechanisms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 359
4.1. Mediated electron transfer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360
4.2. Mediator- less electron transfer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 360
5. Microorganisms in MFCs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 361
6. MFC Design and configurations applied to wastewater treatment. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 362
6.1. Types of MFC design . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 362
6.1.1. Double chamber MFC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 362
6.1.2. Single chamber MFC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 362
6.1.3. Stacked MFC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 363
6.1.4. Flat plate MFC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364
6.1.5. Cylindrical MFC . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 364
6.2. Types of wastewaters applied MFC systems for energy generation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365

Abbreviations: BFC, Biological fuel cell; BOD, Biological oxygen demand; COD, Chemical oxygen demand; CW-MFC, Constructed-wetland microbial fuel cell; DET, Direct
electron transfer; DS-MFC, Desalination microbial fuel cell; EPS, Extracellular polymeric substances; FPMFC, Flat plate microbial fuel cell; MBR-MFC, Membrane-bioreactor
microbial fuel cell; MET, Mediated electron transfer; MFC, Microbial fuel cell; MFM, Microfiltration membrane; NAD, Nicotinamide adenine dinucleotide; PEM, Proton exchange
membrane; PVC, Polyvinyl chloride; SMFC, Sediment microbial fuel cell; UFM, Ultrafiltration membrane; UMFC, Up-flow microbial fuel cell
⁎
Corresponding author.
E-mail address: burcu.gunes@dcu.ie (B. Gunes).

https://doi.org/10.1016/j.psep.2022.03.039
0957-5820/© 2022 The Author(s). Published by Elsevier Ltd on behalf of Institution of Chemical Engineers.
CC_BY_4.0
A. Nawaz, I. ul Haq, K. Qaisar et al. Process Safety and Environmental Protection 161 (2022) 357–373

7. Factors affecting MFC performance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365

7.1. Electrode and membrane material . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365
7.1.1. Anode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 365
7.1.2. Cathode . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 366
7.1.3. Separating membranes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 367
7.2. Effect of pH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 367
7.3. Temperature. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 367
7.4. Substrate loading rate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 368
7.5. Biofilm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 368
7.6. External resistance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 368
8. MFC based integrated process . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 368
9. Future prospects and conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 369
CRediT authorship contribution statement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 369
Declaration of Competing Interest . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 369
Acknowledgments. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 369
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 369

1. Introduction
Booming urbanization with the advent of industrial revolution
has marked an exponential increase in energy consumption across
the world. The surge in the energy demand had insisted on the
utilization of non-renewable energy sources. However, their detri­
mental impacts on the environment and their slow depletion have
shifted the focus to renewable sources that promote sustainability.
Recently, fuel cells are employed as renewable energy sources that
convert the chemical energy within the bonds into electrical energy.
Microbial fuel cells (MFCs) are the unique subset of biological fuel
cells (BFCs) that assured their potential in the generation of bioe­
lectricity, wastewater treatment as well as bioremediation of heavy
metals. It converts the chemical energy embedded in organic or
inorganic matter into electrical energy via a series of electrochemical
reactions (Karthick and Haribabu, 2020; Zhang et al., 2020).
MFC comprises of anodic and cathodic chambers mainly that are
parted by proton exchange membrane (PEM) or salt bridge.
Microorganisms are employed as biocatalysts that oxidize the or­
ganic substrate at anode sequestering protons and electrons.
Electrons are directed to the cathodic chamber via an external circuit
and protons reach the cathodic chamber through the PEM.
Consequently, the successful reduction reaction occurs in the
cathodic chamber by the combination of protons and electrons with
oxygen to yield water (Rabaey and Verstraete, 2005; Enamala et al.,
2020). Microorganisms used as biocatalysts have striking char­
acteristics that have earned them the name of exoelectrogens. As
they have the ability to mediate electrons to the anode surface and
catalyze the reduction reaction of electron acceptor (oxygen) at
cathode (Enamala et al., 2020; Jatoi et al., 2021).
To date, the MFCs have not attained the ideal performance due to
certain shortcomings such as short life span, low production rate
(Clauwaert et al., 2008), membrane fouling (Daud et al., 2019), high
cost and limited efficiencies (Do et al., 2018; He et al., 2017). There
are several parameters that influence the MFC functioning for ex­
ample type of electrode material, temperature, pH, biofilm, loading
rate of substrate, external resistance, and membrane material. Dif­
ferent membrane fabrication techniques can be employed in order to
tailor-make the PEMs for specific purpose MFCs also (RS et al., 2020).
These factors need to be optimized in order to get maximum bioe­
lectricity output (Aghababaie et al., 2015; Butti et al., 2016; Gadkari
et al., 2020). The reactor design also has a great impact on the per­
formance of MFC. Various designs of MFCs have been constructed
that depend upon their mode of operation and the number of
chambers. Primarily, single chamber, dual chamber, and stacked
chamber MFCs are used that are further used in combination to
enhance the performance of MFC (Bhargavi et al., 2018; Flimban
et al., 2018; Nawaz et al., 2020).
Organic wastewater is the one of the most suitable substrates for
bioremediation since it is nutrient rich and yearlong abundance. It
comprises of effluents taken from municipal, industrial, and other
sources that are considered as the prime source for harvesting en­
ergy for power generation (Tharali et al., 2016a, 2016b). Conven­
tional wastewater treatment technologies had major challenges of
high operational cost and energy consumption along with environ­
mental pollution. The cost of traditional wastewater treatment
technologies was estimated to be about the 3% of global electricity
demand in which effluent disposal (i.e., sludge disposal) accounts for
the 50% of the total wastewater treatment cost (Saba et al., 2017; Ye
et al., 2019). Moreover, ineffective conventional wastewater treat­
ment results in the emission of greenhouse gases and other harmful
dissolved substances like phosphates and ammonia (Li and He,
2014). Microbial fuel cells are the key solution to combat these
complications. It plays a crucial role by biodegrading the organic
matter in the wastewater and reducing chemical oxygen demand
(COD) thereby promoting environmental sustainability, low energy
consumption, and cost by nullifying effluent disposal. Several studies
state that the efficiency of COD removal by MFC is approximately
80–90% (Catal et al., 2008; Gadhamshetty et al., 2013; Cui et al.,
2014; Ye et al., 2019). Wastewater from various sources has been
treated to produce power output (Hu, 2008; Aldrovandi et al., 2009;
Sangeetha and Muthukumar, 2011; Thung et al., 2015).
MFCs are environment friendly alternatives to wastewater
treatment and bioelectricity generation however, they have certain
limitations. Scaling up MFC process is a major challenge that further
increases the overall cost and power consumption. Therefore, more
research is required in this regard to introduce cost-effective MFC-
based integrated technologies that would pave the way to sustain­
ability (Nawaz et al., 2020). To date, no review connects wastewater
treatment with energy production by employing different types of
reactor designs of microbial fuel cells to the best of authors’
knowledge. Therefore, this review fills the gap by highlighting the
best type and design of MFC so far. This is achieved by drawing
comparisons among different types of microbial fuel cells (double/
single chambered or parallel/series) and waste water sources. Sub­
sequently, progressing towards a greener approach that is MFC-

358
A. Nawaz, I. ul Haq, K. Qaisar et al.
Fig. 1. Working of a Typical Microbial fuel cell for electricity generation.
based integrated/hybrid technologies. Thus, it includes an overview
of MFC design, mechanism, types of wastewaters, and microorgan­
isms present in wastewater along with the factors affecting MFC
performance of generating electricity. An effort is made to confer the
current and emergent areas to the best of the authors' ability.
2. What are microbial fuel cells (MFCs)?
MFC is a device operated on the basis of electrochemical energy
conversion, in which microorganisms can generate electro­
chemically active substances. These substances react on anode to
form a biofilm which acts as a biocatalyst to produce electricity
(Allen et al., 1993). It consists of two chambers made of poly­
carbonate, plexiglass or glass with respective electrodes of carbon
cloth, graphite, and carbon paper etc. One named as anodic chamber,
which is an anaerobic part of the MFC, nitrogen gas is used to
maintain this anaerobic condition. Wastewater comprising of dif­
ferent types of microorganisms will be treated by electrochemical
means in this chamber. The other is a cathodic chamber which is an
aerobic part consisting of a high potential electron acceptor elec­
trode usually oxygen. Usually, a proton exchange membrane made of
Nafion and Ultrex is used to separate the two chambers. They have
an effective role as a membrane for PEM fuel cells by allowing ca­
tions to pass through them (Zhou et al., 2018). Fig. 1 displays a
diagram of a typical MFC.
3. Bioelectricity generation from wastewater via MFC
Wastewater is an effective source of renewable energy storage.
Microbial fuel cell is used simultaneous wastewater treatment and
cost–effective electricity production. Wastewater is utilized as a
substrate originates from either human, animal, or food processing
sources. In a typical MFC, electrical energy potential of the organic
pollutants present in wastewater in form of carbonaceous com­
pounds is recovered via biochemical conversion system that gen­
erates electricity (Tatinclaux et al., 2018). MFC, due to its renewable
biomass property is an ideal technology for bioelectricity produc­
tion. Microorganisms are present in wastewater that act as bioca­
talysts to oxidize organic or inorganic matter. They metabolize
organic matter for energy creation and growth while generating
electrons, protons, and CO2.y creation and growth.
Microbial reduction results in the production of electrons that
are attracted towards the anode. They attach to anodic surface with
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Process Safety and Environmental Protection 161 (2022) 357–373
the help of cytochromes or redox-active proteins via a biochemical
catalyst reaction. Electrons will travel towards the cathode by an
external circuit (usually a copper wire). Meanwhile, protons pro­
duced by the microorganisms will travel towards the cathode
through PEM. Protons will produce H2 (Hydrogen gas) and hydroxyl
(OH-) in cathode chamber and electrons will be reduced to pure and
clean water (H2O) by reacting with a strong oxidizing agent like
oxygen (Hamelers et al., 2010). Ferricyanide is also an oxidizing
agent that is less commonly used in the cathodic chamber (Ucar
et al., 2017). Oxygen reduction at cathode is catalyzed by precious
and non-precious metal, platinum and palladium respectively
(Cheng et al., 2006; Dombrovskis et al., 2016; Kumar et al., 2015).
Following are the equations that depict the overall reaction and
reactions taking place at anodic and cathodic chambers singularly.
Anodic reaction: CH3COO- + H2O 2CO2+8H+ + 8e-
Cathodic reaction: 8H+ + 8e- + 2O2 4H20
Overall reaction: CH3COO- + 2O2 2H2O +2CO2
The water produced in MFCs can be further utilized to move the
turbines for bioelectricity production at a commercial scale. Stored
water is passed through the tunnels and falls on the turbines. The
shaft of the turbine then rotates the generator to produce electricity.
Water is transported to the turbine through a pressure pipe known
as penstock. During installations, a valve must be kept at the top of
penstock which can be shut when the turbine is required to be shut
down and evacuated for maintenance (Paish et al., 2002).
4. Electron transfer mechanisms
Electron transfer occurs as biomolecules present in bacteria, like
proteins, mediate in donating and accepting the electrons between
the bacteria and electrodes. There are two main mechanisms by
which extracellular electron transfer takes place, Direct Electron
Transfer (DET) or mediator-less electron transfer and indirect elec­
tron transfer or Mediated Electron Transfer (MET) (Kumar et al.,
2018). Two bacteria, Geobacter spp. and Shewanella spp., have been
studied extensively for extracellular electron transfer mechanisms
(Kumar et al., 2016). Fig. 2 exhibits the schematic representation of
electron transfer mechanisms.
A. Nawaz, I. ul Haq, K. Qaisar et al.
Fig. 2. Schematic representation of Electron transfer mechanism in MFC.
In mediated electron transfer MFC, microorganisms have no
electrochemically active proteins on their surface for transferring
electrons to the anode. Consequently, they require agents called
mediators or electro-active metabolites to transfer these electrons
between the electrode and microorganisms. These mediators at
higher concentrations increase the availability of the transferred
electrons. However, depending on the microorganism involved,
mediators may be synthetic, like neutral red, or natural, like sulfate/
sulfide (Rahimnejad et al., 2015). They cross the outer cell lipid
membrane and plasma wall and start liberating electrons, which are
usually taken up by oxygen and other intermediates. Subsequently,
the reduced mediator exits the electron-loaded cell and deposits the
carried electrons on the surface of the anode electrode. Hence, the
anode electrode becomes negatively charged. Upon releasing the
electrons, the mediator returns to its original oxidized state pre­
pared to repeat the process. On contrary, direct electron transfer
doesn’t require any mediator making them cheaper and non-toxic.
Mediator-less MFC function without exogenous electron carriers.
Instead, their electron transport relies mainly on metal-reducing
bacteria therefore it is vital in treating wastewater containing heavy
metals (Katz et al., 2003).
Factors to be maintained for efficient electron transfer include
the presence of electrochemically active enzymes, circuit external
resistance, and biofilm production at anode. Moreover, proper
transfer of the protons across the membrane must be assured as
inappropriate transport can reduce the current. As limited proton
supply would lead to slow cathode reaction and alteration in pH
consequently restricting the electron transfer and microbial activity.
In terms of optimum energy production, mediator-less electron
transfer is preferred over mediated one since it is safer and cheaper
and produces maximum power (Gil et al., 2003).
4.1. Mediated electron transfer
Many microbes lack the ability of direct electron transfer either
due to absence of surface-active proteins or because they have no
direct contact with the electrode surface (Reguera et al., 2005). In
such cases, they use an indirect means to transfer electrons. MET
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Process Safety and Environmental Protection 161 (2022) 357–373
usually includes a redox exporter that acts as a shuttle to transfer
electrons to the terminal electron acceptor. In MFCs, these redox
mediator gains electrons from bacterial cells, leaving them in a re­
duced state, and transfers electrons to the anode, get oxidized and
then free to initiate a subsequent round of electron transfer.
Ieropoulos et al. (2005), enlisted properties of good mediator in­
clude: (1) ability to cross the bacterial cell membrane easily, (2) non-
toxicity towards bacteria, (3) good solubility in the anolyte, (4) suf­
ficiently positive redox potential to facilitate electron transfer, (5)
low cost and easy commercial availability.
Redox mediators can either be exogenous (externally added
compounds), or endogenous (produced by the bacteria themselves).
Endogenous mediators are usually metabolites produced by the
cells, like flavins (Marsili et al., 2008), ribofavins (Reguera et al.,
2006), phenazines (Glasser et al., 2017), and quinones. Electro­
chemical studies of S. oneidensis MR-1 have confirmed the presence
of secreted flavins participating in electron transfer (Marsili et al.,
2008). Removal of riboflavins from biofilms of S. oneidensis MR-1
decreased electron transfer to electrodes by over 70% (Marsili et al.,
2008). Accordingly, it was proposed that flavin molecules bind to the
outer membrane cytochromes for electron transfer (Slate et al.,
2019). Whereas usage of exogenous mediators was the norm
through the initial years of MFC research, as exoelectrogenic mi­
crobes were not yet known (Akiba et al., 1987; Park et al., 1999).
Exogenous mediators used to enhance MFC performance include
neutral red, methylene blue, thionine, Fe (III) EDTA and various
quinone derivatives (Vega and Fernández, 1987; Park and Zeikus,
2000). The use of exogenous mediators has significantly reduced
over the years due to their instability and toxicity yet, this technique
is still comparatively common to improve MFC performance.
4.2. Mediator- less electron transfer
Mediator-less or direct electron transfer mechanism involves
transfer of electrons directly from the microbes to the electrodes
without the aid of any shuttle or mediator. For this purpose, a
physical contact between the bacterial cell membrane and electrode
is necessary (Lovley et al., 2006). Direct electron transfer is achieved
A. Nawaz, I. ul Haq, K. Qaisar et al.
by outer membrane cytochromes (OMCs), conductive pili or self-
assembled appendages called nanowires as shown in Fig. 2 (Reguera
et al., 2018). C-type cytochromes, present on outer surface of bac­
terial cell act as redox proteins and assists in transfer of electrons to
terminal electron acceptor. C-type cytochromes are multi-heme
proteins that have a redox potential range nearly 1 Volt and are
rationally stable against chemical modifications (Liu et al., 2014).
According to a study by Holmes et al. (2006), on Geobacter sulfur­
reducens, where an electrode was provided as the single electron
acceptor and a 19-fold increase in transcripts of OMCs was observed,
which was responsible for encoding OMCs required for metal re­
duction. Concluded that Geobacter and Shewanella contain intricate
networks of OMCs participating in extra cellular electron transfer
thereby contributing to higher current densities in MFCs.
Another way of forming direct electron transfer is the estab­
lishment of conductive pili called nanowires in exoelectrogenic mi­
crobes that allow the microbes to transfer electrons to a solid
electron acceptor (Reguera et al., 2005). These nanowires are linked
to cytochromes present on bacterial membrane and allow electron
transfer to terminal electron acceptors (TEAs) that are physically
distant from the microbes (Reguera et al., 2018). While flagella and
pilli are homopolymers comprising of a single subunit, nanowires
are an assemblage of different cytochromes and outer membrane
proteins (Sure et al., 2016). Three types of nanowires have been
elucidated as of now: nanowires made up of type IV pili (Li and Li,
2014), nanowires consisting of periplasmic proteins and outer
membrane cytochromes (e.g., MtrC, Omc A) (Pirbadian et al., 2014).
Type IV nanowires are mainly found in G. sulfurreducens and Sy­
nechocystis; nanowires comprising of MtrC and Omc A are observed
in Shewanella oneidensis (Sure et al., 2016).
5. Microorganisms in MFCs
Microorganisms play a central role in generating energy in a
microbial fuel cell as they carry out the biochemical reactions. MFCs
employ two types of microorganisms such as electrode reducing and
oxidizing. Electrode reducing microorganisms carry out the reduc­
tion of electrodes and electron movement towards anode whereas
microbes involved in the oxidation of organic components and their
subsequent transfer towards electrode called electricigens are elec­
trode oxidizing (Fan and Xue, 2016). Electrons can be passed either
by a mediator molecule in the solution, by proteins in the outer
membrane, or through pili that are present at the outer surface of
bacteria. On contrary, electrode oxidizing microorganisms are those
that receive electrons from the cathode and reduce various sub­
stances like acetate and carbon dioxide. The selection of micro­
organisms must be done keeping in mind their capacity to transfer
electrons from substrate to anode (Nawaz et al., 2020).
Microorganisms used for electricity generation in MFC can either
be pure cultures or mixed cultures (Shehab et al., 2017). Pure cul­
tured microorganisms such as Geobacter (Lovley et al., 1993) and
Shewanella (Gorby et al., 2006) species are predominant among
microbial population that have been used in MFC technology. Var­
ious other microorganisms that can play a significant role in pro­
ducing electricity are yet to be discovered. Among pure cultured
microbes, different types of microbes have been employed for power
production in microbial fuel cell. List of some potent eletricigens are:
[1] Archaebacteria, many archaea have the ability to withstand ex­
treme conditions like high temperature and salinity. They are es­
sential in generating electricity under certain conditions (Cao et al.,
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Process Safety and Environmental Protection 161 (2022) 357–373
2019). [2] Acidobacteria, a diverse acidophilic group of bacteria that
is present in various environments. They have the ability to consume
a vast variety of substrates (Dedysh et al., 2018). [3] Cyanobacteria
are environmentally friendly and photosynthetic microorganisms
having applications in MFCs. They produce electricity, by oxidation
of water using light as a source of energy (Indrama et al., 2019). [4]
Proteobacteria constitute the major group of electricity producing
bacteria used in MFC. Most of its strains can transfer electrons di­
rectly to the electrodes. Various species belonging to the classes; α-
proteobacteria, β-proteobacteria, γ-proteobacteria and δ-proteobacteria
are potent electricigens. Little research has been made on using
eukaryotes in MFCs. Among eukaryotic microorganisms, Yeasts are
highly beneficial in producing electricity due to their rapid growth
rate. However, it still generates lesser energy than bacterial strains.
Progress has been made by engineering its strains that yield greater
energy (Cao et al., 2018). Furthermore, Microorganisms of genera
Geobacter (Reguera et al., 2006), Shewanella (Kim et al., 2002),
Pseudomonas, and Clostridium (Park et al., 2001) are worthy exoe­
lectrogens.
Mixed Cultures of microorganisms are utilized to improve bio­
logical constancy by obtaining them from extensively used sources
including soil or marine sediment, natural microbial community,
and brewery wastewater (Munoz-Cupa et al., 2020). Various other
sources of microorganisms including domestic water, activated
sludge, metal reducing, municipal wastewater, and various other
kinds of wastewater have been considered for microbial fuel cell
(Cecconet et al., 2018; Chen et al., 2020). A number of microbial
species are present in wastewater that can produce electricity. In this
regard, wastewater is used as an inoculum in MFC. Recently, pho­
tosynthetic bacteria have also been studied to serve as an effective
source of electric power generation. This technique has proved to be
environment friendly as it eliminates carbon dioxide from the at­
mosphere thereby coupling the process of photosynthesis and
bioelectricity generation (Rosenbaum et al., 2005).
Another emerging method for electricity generation is the de­
velopment of a synergistic association between heterotrophic bac­
teria and photosynthetic bacteria. This method is based on a
symbiotic relationship that involves the consumption of organic
matter that is synthesized as a result of photosynthesis of hetero­
trophic bacteria. Strains of Pseudomonas aeruginosa are used to­
gether with the utilization of co-factor NAD, resulting in the surge of
metabolic rate and bacterial potential consequently leading to
greater generation of bioenergy (Tharali et al., 2016a, 2016b).
Bacteria form biofilm to orient themselves at the anode in order
to accomplish the oxidation of substrate, using anode as receiver of
electrons. These electroactive bacteria in a biofilm produce in­
tracellular components forming a matrix so that they adhere to
anode. It is composed of bacterial cells, extracellular proteins, and
sugars that help bacteria in transporting electrons to anode surface
(Marcus and Rittmann, 2008). Biofilm formation also occurs by the
electrostatic interaction between organic compounds harboring
negative charges in nutrient buffer and debris that bacteria produce.
In addition to this, the extracellular polymeric substances produced
by the microorganisms are an integral part of biofilms. These (EPSs)
have a high molecular weight and influence different physico­
chemical processes like the transfer of electrons (Gunes, 2021;
Marsili et al., 2008). Fig. 3 sketches the stages for biofilm formation
at anode.
A. Nawaz, I. ul Haq, K. Qaisar et al.
Fig. 3. Mechanism of biofilm formation in MFC.
6. MFC Design and configurations applied to wastewater
treatment
MFC performance not only depends upon the biocatalysts but
also on the design of the reactor. The key elements of the fuel cell are
electrodes, PEM or salt bridge and external circuit (resistor). The
design will directly affect the electricity production, budget, and also
the efficiency of the cell. Several factors like shape, size, electrode
type, and stacking direction of MFC are required for efficient de­
signing of MFC (Gul et al., 2021). Depending on the design of the
reactor there are various types of MFCs. Different MFC configura­
tions is discussed in the following sections in detail. Generally, MFC
model can be either a single or double chamber, additionally, it can
also be used in combination (Pant et al., 2010). The power outputs of
different MFC configurations are shown in Table 1. Double chamber
MFCs are mostly used as they have membranes for ion exchange,
two separated chambers and high energy output. Although, the
single chamber has been evolved from the double chamber, how­
ever, it has more encouraging results (Call et al., 2008). Fig. 4 shows
the diagrammatic representation of different MFCs employed to
treat wastewater.
6.1. Types of MFC design
6.1.1. Double chamber MFC
Double chamber MFC consists of two compartments, anodic and
cathodic which are anaerobic and aerobic respectively. Both regions
are connected externally through a wire and internally through the
proton exchange membrane. This whole frame completes the circuit
and membrane facilitates the protons to reach the cathodic chamber
by limiting the oxygen diffusion in the anodic chamber (Min et al.,
2004; Du et al., 2007; Rahimnejad et al., 2015; Slate et al., 2019; Xia
et al., 2018). The submerged electrodes can be made of any non-
corrosive conductive materials. This runs in batch mode with a
chemically defined medium to generate power. An external re­
sistance directly affects the output, as the long distance between the
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Process Safety and Environmental Protection 161 (2022) 357–373
electrode and cathode would cause a decrease in output (Logan
et al., 2005; Choi et al., 2013). For this internal resistance is de­
creased from 672 Ω (103 mW/m2) to 93 Ω (57 mW/m2) and elec­
trodes were placed close enough to increase the power density.
Double chamber MFC is mostly applicable for wastewater treat­
ment to produce electricity on large scale (Min et al., 2004). How­
ever, cathode needs consistent solution replacement with aeration
to provide oxygen. Nevertheless, the power production can still be
scaled up by using it by controlling the pH of cathode, purging fresh
O2, increasing flow rate, adding additional electron mediators, and
maintaining the internal resistance.
Generally, a dual chambered microbial fuel cell functions in the
water cathode mode. This distinctive design can be altered into
different designs; single chamber, stacked, and cylindrical mode
MFCs. Therefore, MFC configuration is a key factor in determining
the power production to achieve greater efficiencies and reduced
costs for industrial applications.
6.1.2. Single chamber MFC
It is employed for power production at a small scale with less
internal resistance thereby promoting low cost with high efficacy.
The design is mainly based on double chamber MFC with few
changes, there is only one defined anodic chamber, where waste­
water is placed (An et al., 2014). Single chamber MFC has no defined
cathodic chamber and PEM, only a porous cathode is present on one
side that allows protons to pass through by restricting the flow of
oxygen towards the anode (Liu and Logan, 2014). On the other side,
oxygen is captured from the atmosphere to complete the circuit.
Whereas anodes are normal carbon electrodes, cathodes are either
PEM fused with flexible carbon cloth or porous carbon electrodes.
Nonetheless, graphite covering is frequently used for cathodes.
Usually, electrolytes are dispensed into cathode in a steady state and
act like a catholyte preventing the cathode and membrane from
drying out (Cheng et al., 2006).
Single chamber MFC allows air enriched with oxygen to directly
react with porous cathode electrode. It is operated in air-cathode
A. Nawaz, I. ul Haq, K. Qaisar et al. Process Safety and Environmental Protection 161 (2022) 357–373

mode. The benefits of single-chamber MFCs include less internal

(Aelterman et al., 2006; Rodrigo

(P. Zhang et al., 2017; L. Zhang

resistance, improved proton diffusion, cheaper, higher oxygen re­

(Subha et al., 2019;Arvaniti

(Tran et al. (2019);Min and
duction rate at the cathode, simple to use, and aeration making it
more adaptable. However, several constraints limit its use at com­
mercial scale such as evaporation, high-rate oxygen diffusion and
(He et al., 2016)

liquid outflow (Liu and Logan, 2004).

Logan (2004)
et al., 2007)

et al., 2021)

et al., 2017)
Reference

6.1.3. Stacked MFC

MFCs can be stacked either in parallel or series and they play a
key role in enhancing power production. Stack configuration yields
2.22 W/m3 (series)
higher current output if placed in parallel and high voltage output if
Power density

placed in series connection. The coulombic efficiency is not influ­

679.7 mW/m

175.7 W/m3
25 mW/m2

98 mW/m2

enced by design; however, the power of overall battery is amplified

(parallel)
1.1 W/m3

(Flimban et al., 2018). Multiple factors must be kept in view while

treating wastewater with stacked MFCs capable of generating en­
ough power. The factors include maintaining the substrate flow rate,
height if MFC is vertically stacked, check organic concentrations at
Need regular solution replacement in cathodic chamber

end of MFC operation, and volume of the reactor (An et al., 2015).
These have not been investigated in detail, so several
Costly due to high energy demand for fluid pumping

factors like height, shape, direction of flow are to be

Stacked MFC is further sub-divided into horizontal stack and vertical

Membranes are damaged when used consistently.

stack.
A researcher group concluded that high current density is
Long distance among anode and cathode,

achieved in parallel connections. It is also deduced that surface

Not suitable for large scale production

material of electrodes is directly affects COD removal and electricity

Membrane permeability to oxygen.

production. Table 2 shows the performance of various MFC stacks

with different conformations cited in the literature.

6.1.3.1. Horizontal stacked. They are rectangular, parallel piped

double chambered MFCs with horizontal stacking. They comprise
Disadvantages

of anode, PEM and cathode inside a rectangular frame having two

ports; one inlet and one outlet (Wu et al., 2016; Aelterman et al.,
observed

2006; Vilajeliu-Pons et al., 2017; Rahimnejad et al., 2012; Jafary et al.,

2013; Zhang et al., 2017). Horizontally stacked MFC requires pumps
to circulate the wastewater. They are linked to current accumulators
Increased fluid recirculation results in greater COD

with metal wires thus making the circuit in parallel or series. For
High rate of diffusion of electrons and protons.

removal, works in continuous operation mode,

partitioning the horizontal stacks, acrylic sheets or rubbers are also

Easy to scale-up and have high loading rate.

used in order to avoid catholyte and anolyte sharing within the MFC
Reduce set-up costs, higher power output

Used for scaling up the power production

stack unit.
Aelterman et al. (2006) evaluated the power density for six
Short distance between electrodes.

horizontally stacked MFC units on the basis of polarization curves.

Power could be as high as 263 W/m3 for parallel connections and
308 W/m3 for series connections. Recently, conferring to (Wu et al.,
Potential of bioelectricity generation with respect to MFC types and configurations.

2016), the pilot project attained a highest power density of 51 W/m3

Scalability of MFC

and COD removal efficiency of 97% from their scaled-up, 72 Liter,

rectangular, horizontal-stacked parallel-connected MFC.
Advantages

6.1.3.2. Vertical stacked. Vertical stack MFC is based on parallel

electrode connections in series flow mode. They are inexpensive
because of their self-driven mechanism under the force of gravity.
Cathode: PEM bonded with flexible

Carbon paper anode electrode and

These don’t need pumps to maintain the flow rate. Many successful
carbon cloth cathode electrode.

events were recorded using vertical stacked MFC. The first self-
Graphite –cement composite

sustainable MFC stack capable of self-maintenance used a vertical,

Carbon cloth electrodes

single-chambered MFC cascade consisting of 40 identical 20 mL

Anode: Graphite fiber.

units (total volume of 0.8 L) (Ledezma et al., 2013). However,

Carbon electrodes

power generated by vertical MFC stack was low (110 m/W with
carbon cloth

maximum power density of 10 W/m3). The voltage reversal in

Electrodes

stacked microbial fuel cells is a challenge that must be considered,

and the information on its certain cause and existence is however,
doubtful (An and Lee, 2014).
Therefore, a number of aspects must be considered while de­
Double chamber
Single chamber

termining if a vertical cascade MFC has capacity of both generating

enough power and adequately treating wastewater: (1) the height of
Flat plate

Up flow

Stacked

the MFC system if it is gravity-fed, (2) the method required for

Types
Table 1

maintaining the substrate flow rate (gravity-fed vs. pumps), (3) the
volume of the reactor (Flimban et al., 2019).

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A. Nawaz, I. ul Haq, K. Qaisar et al. Process Safety and Environmental Protection 161 (2022) 357–373

Fig. 4. Diagrammatic representation of different types of MFCs.

6.1.4. Flat plate MFC
The cubic, mini-squared flat compact plate microbial fuel cell
(FPMFC) is the system resembling the chemical fuel cell having
cathode and anode attached through PEM internally and externally
through the wire made of copper or any other conductive material
(Du et al., 2007; Janicek et al., 2014; Min and Logan, 2004). In FPMFC,
the cathode is hot pressed to Nafion proton exchange membrane
making a PEM assembly where anode is attached on the other side of
the membrane. In cubic FPMFC, anodic chamber is supplied with
wastewater and the cathodic chamber is provided with air without
any catholyte. Major difference between the cubic MFC and other
type is less internal resistance due to small distance between the
hence increasing the power output (Janicek et al., 2014).
Kim et al. (2007) conducted an experiment and tested cubic
shaped MFC performance against bottle shaped MFC. It was seen
that former produce 14 times high power generation (214 mW/m2)
with 84 Ω internal resistance than that of the later (38 mW/m2) with
1272 Ω internal resistance.
6.1.5. Cylindrical MFC
Cylindrical MFC also called Up-Flow MFC is used to treat was­
tewater rather than producing electricity at a large scale. It is oper­
ated in continuous or series mode to enhance power production. Up-
flow microbial fuel cell has uniform graphite electrodes with proton
exchange membrane (PEM), operated in fed-batch mode. Various
configurations of cylindrical MFC were tested having different dis­
tance between their electrodes that directly affects the internal

Table 2
Comparative analysis of power generation on the basis of stacked MFC types and their configurations with respect to (s): Series connection; (p): Parallel connection.

Conformations No. of MFCs Electrodes Internal resistance (Ω) Maximum power density W/m3 Reference

Two chamber MFC stack
Two chamber MFC stack
Two chamber MFC stack
Single chamber MFC stack
6 Graphite granules 6.5 (series) 0.085 (s) (Aelterman et al., 2006)
0.425 (p)
3 Graphite 11.5 Ω/m2(series) 0.11 W/m2 (s) (Jafary et al., 2013)
1 Ω/m2 (parallel) 0.13 W/m2 (p)
4 Carbon cloth 11.5 Ω/m2(series) 2.22 W/m2 (s) (P. Zhang et al., 2017; L. Zhang et al., 2017)
1 Ω/m2 (parallel) 1.98 W/m2 (p)
10 Carbon fiber veil 11.5 Ω/m2(series) 0.97 (p)
1 Ω/m2 (parallel)

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A. Nawaz, I. ul Haq, K. Qaisar et al. Process Safety and Environmental Protection 161 (2022) 357–373

Table 3
Comparative potential of different types of wastewaters to generate power density and COD removal (%).

Type of Wastewater COD Removal (%) Power Density References

Water from fermentation processes with different end products 85–90 62–835 mW m-2
Wastewater rich in carbohydrates with acetate and butyrate as main elements 98 305–506 mW m-2 (Liu et al., 2005)
Acidogenic food waste leachate 87 432 mW m-2 (Li et al., 2013)
Chocolate industry 75 1.5 mW m-2 (Patil et al., 2009)
Brewer wastewater 20.7 669 mW m-2 (Wen et al., 2010)
Petroleum refinery 64 330.4 mW m-2 (Guo et al., 2016)
Starch processing 98 239.4 mW m-2 (Lu et al., 2009)
Food processing 86 230 mW m-2 (Rabaey et al., 2005)
Cooking 50 538 mW m-2
Dairy industry 90.46 621.13 mW m-2
Simple wastewater 77 511.11 mW m-3 (Touach et al., 2016)
Winery 8.459 890 mW m-2 (Penteado et al., 2016)
Tannery 88 7 mW m-2 (Sawasdee et al., 2016)
Industrial 84 131 mW m-3 (Touach et al., 2017)
Domestic 77.9 90 Wm-3 (Jiang et al., 2013; Rabaey et al., 2005)

wastewater (a complex substrate). In contrast, complex organic

Fig. 5. Flowchart of parameters depicting MFC performance for bioelectricity gen­
eration.
resistance. The substrate, wastewater enters the anodic chamber at
the bottom, processed and electrons are delivered to the cathode
with help of an external circuit. Meanwhile pumping and re­
circulation of fluids is required in cylindrical/Up-flow MFC, so large
volumes of wastewater can be treated but power production is af­
fected due to greater energy loss. For this purpose, we use MFC in
fed-batch or continuous mode to enhance power production (He
et al., 2005). Recirculation mode is effective in fed-batch for effective
mass transfer and electricity production. Whereas, in continuous
mode, hydraulic retention time decreases thus increasing the power.
However, we can avoid clogging in this mode by elimination of en­
tire suspended solids (Jiang et al., 2011).
He et al. (2005) displayed two configurations of UPMFC, one
presenting cathode and anode separated by certain distances and
having 84 Ω internal resistance with maximum power density of
170 mW/m2. The other configuration giving low internal resistance
as electrodes are placed near to each other resulting in a power
density of 29.2 W/m2. Thus, it concludes that a smaller distance will
generate more power in short period of time.
6.2. Types of wastewaters applied MFC systems for energy generation
Wastewater collected from municipal, industrial, and various
other sources can efficiently be used as a substrate for producing
bioenergy in microbial fuel cell. Performance of MFC for treating
wastewater and power output can be evaluated with reference to
maximum power density, coulombic efficiencies, and COD. By using
pure substrates like acetate, sucrose and glucose, the maximum
power densities recorded are high compared to that of the
substrates are problematic as they interfere with electricity gen­
eration. Further, production of volatile acid and high concentration
of ammonia leads to toxicity during fermentation of substrate (Min
et al., 2005).
The composition and source of wastewater highly influence the
working potential of MFC. The utilization of municipal wastewater
for bioelectricity generation in MFCs has a drawback as maintenance
of stable production of electricity cannot be achieved. This is influ­
enced by the presence of high quantity of chemical oxygen demand
(COD) (Kim et al., 2015). Normally, industrial wastewater has a tre­
mendous variety of nutrients present in it and this composition is
determined by the type of industry from which wastewater is col­
lected (Logan, 2005). The wastewater obtained from a food industry
will contain a high amount of BOD but will not contain any toxic
compounds. similarly, the wastewater collected from a brewery in­
dustry will be a rich source for energy production as it contains
proteins, sugars, and starch that can be used by microorganisms
(Wen et al., 2010). Whereas distillery wastewater is characterized by
high salt concentration and organic load (Mohanakrishna et al.,
2010). Consequently, a suitable construction must be designed for
the biological treatment process keeping in view the composition of
wastewater. Table 3 enlists several wastewater types that have been
used as MFC substrate in various studies, along with their COD re­
moval (%) and power density.
7. Factors affecting MFC performance
Microbial fuel cell is considered as one of the promising tech­
nological innovations for concurrent wastewater treatment and ef­
ficient generation of electricity (Bose et al., 2019). MFC performance
and electricity generation are influenced by various factors such as
electrode and membrane material, temperature, pH, substrate
loading rate, biofilm, and external resistance. In order to improve the
energy, output these factors must be optimized. Fig. 5 shows the
web diagram of factors affecting MFC performance and electricity
generation and the impact of each factor is discussed in detail in the
following sections.
7.1. Electrode and membrane material
7.1.1. Anode
The anode compartment is one of the vital parts of microbial fuel
cell device as the production of electrons takes place at anode and

365
A. Nawaz, I. ul Haq, K. Qaisar et al.
therefore, it is referred to as the heart of MFC. In addition to the
production of electrons, oxidation of organic pollutants to generate
protons together with electrons, formation of biofilm, and produc­
tion of hydrogen occur in anode compartment. In order to enhance
the MFC performance, the selection of suitable material for anode is
imperative. The material employed should possess high electrical
conductivity, compatibility with microorganisms, chemical stability,
good surface area, and low cost (Jaiswal et al., 2020). Compatibility
with microbes of anode material is crucial. Many materials such as
gold, copper, stainless steel platinum show characteristically high
electrical conductivity but their use as anode material is limited as
they are highly expensive and inhibit cellular growth by anti­
microbial substances such as cisplatin formed from platinum
(Munoz-Cupa et al., 2020; Slate et al., 2019). Alternatively, highly
compatible and low cost carbonaceous anode materials have been
employed including graphite rod, graphite fiber brush, graphite
plates, carbon fiber, carbon cloth, carbon paper, carbon brush, carbon
mesh, carbon felt, carbon rods, and carbon nanotube. Among carbon
materials used, carbon brush, due to its fibrous nature has increased
surface area. Carbon rods are inexpensive and show significant
electrical conductivity but possess a small surface area. Carbon veil,
carbon paper and carbon mesh are highly porous and cost-effective
materials but are fragile for mechanical applications. Carbon felt is
porous having a large surface area and appreciable conductivity
(Mustakeem, 2015; Singh et al., 2019; Jatoi et al., 2020).
Several modifications of anode material have been made to ele­
vate the working potential of MFCs. The introduction of carbon na­
notubes as anode material has helped increase the efficiency of MFCs
as they offer substantial conductance, mechanical stability, and
biocompatibility (Yazdi et al., 2016). A study evaluating the effect of
multi-walled carbon nanotubes on MFC performance and microbial
growth was conducted by comparing anodes modified with
MWCNT-, MWCNT-COOH and MWCNT-NH2 with bare carbon cloth
anode. It was observed that anode modified with MWCNTs showed
denser microbial colonization and the greatest biomass of 39.27
nmol p/cm2 was observed at MWCNT-NH2 modified anode, followed
by 28.44 nmol p/cm2 for MWCNT-COOH, 19.66 nmol p/cm2 for
MWCNT, and 13.23 nmol p/cm2 for unmodified anode. For power
density, the highest value (560.4 mW/m2) was obtained for MFC
with MWCNT-COOH modified anode, implying a 49% increase in
power output than MFC with unmodified anode having power
density 375.58 W/m2. Thus, the study indicated the improvement of
MFC performance using MWCNTs modified anode due to their large
surface area supporting microbial growth and greater electron
transfer potential (Fan et al., 2017).
Application of nitrogen for modification of anode material has
helped enhance the bioelectricity generation potential of MFC.
Modification of anode with nitrogen-doped porous carbon material
has been shown to increase the performance of MFC by promoting
microbial growth and transfer of electrons to the electrode from
microbes (Bi et al., 2018). The use of nanocomposites of various
metals and their oxides for modifying anode improves the efficiency
of MFC by increasing the adherence of microbial cells and lowering
ohmic loss. Oxides of titanium (Ti), iron (Fe), tin (Sn), manganese
(Mn), etc. integrated on carbon-based material forming nano­
composite for modification of anode has also been employed for
improving the efficiency of MFC. Application of conductive polymers
such as Polyaniline for anode modification caught attention due to
their high conductivity potential, enhanced bacterial adherence, and
robustness to environmental conditions (Hindatu et al., 2017;
Mehdinia et al., 2014). Yellappa et al., 2020 investigated the potential
of stainless steel mesh integrated with polyaniline functionalized
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Process Safety and Environmental Protection 161 (2022) 357–373
activated carbon (PANi-FAC) composite as anode. As compared to the
power output of stainless-steel mesh anode (169 mW/m2), SSM-
PANi/FAC resulted in higher power output (322 mW/m2). Similarly,
charge-transfer resistance (Rct) displayed by SSM-PANi/FAC was
29.9 Ω that was considerably lower than SSM having Rct 678 Ω,
suggesting it as an effective anode composite improving biocatalytic
potential with lesser ohmic losses in MFC. Other modifications in
anode for improvement in MFC performance involve treatment of
anode with heat and acid. Heat treatment aids in improving power
density by eliminating the impurities at anode that hinder electrical
conductance (Kalathil et al., 2017). In this regard, Ni et al. (2020)
reported the effect of acid–heat based treated anode carbon cloth on
MFC performance. A comparison of untreated carbon cloth (CC),
acid–heat-modified carbon cloth (N_H_CC), nitric acid-modified
carbon cloth (N_CC), and ferric chloride-modified carbon was made
for their influence on performance of MFC. The acid-heat treated
carbon cloth was prepared by initially oxidizing in 65% nitric acid at
50 ◦C for 6 h followed by heat treatment at 450 °C for 2 h. The
highest power density of 883.62 mW/m2 was observed in MFC with
acid-heat treated anode displaying 350% increment in power density
as compared to unmodified anode. Similarly, the highest COD re­
moval (80.1%) was observed in acid-heat modified anode. Acid-heat
treatment also resulted in selective growth of microbial commu­
nities which was considered to aid in improving MFC performance,
thereby, revealing positive effect of acid-heat treatment on the
properties of anode carbon cloth.
7.1.2. Cathode
Cathode electrode in MFC is responsible for receiving electrons
and protons generated at anode. Double chamber MFC uses cathode
separated by separating membrane whereas single-chamber MFC,
without membrane, utilizes air cathode that receives the generated
electrons. On reaching cathode, the electrons are reduced by the
action of electron acceptors present in cathode (Flimban et al., 2019).
For cathode, different materials including carbon cloth, graphite
fiber brush, graphite rod, carbon paper, graphite felt, and stainless
steel have been used (Zhang et al., 2012; Janicek et al., 2015; Chen
et al., 2018; Cecconet et al., 2018). As an electron acceptor, Oxygen is
highly preferred due to its greater redox potential. However, its use
is limited as it shows insufficient contact with electrode and in­
creased power consumption for its supply. Alternatively, different
electron acceptors, in place of oxygen, have been utilized (Ucar et al.,
2017). Among them are tetrachloroethane, hexavalent chromium,
ferric ions, perchlorate, nitrite, sodium bromated, dichloroethane,
potassium ferricyanide, and fumarate (Jaiswal et al., 2020). Ad­
ditionally, cathode requires catalysts, such as platinum, for enhanced
performance. However, these catalysts are costly, exhibit lesser
sustainability, and require increased preparation time, thus making
the process expensive (Chaturvedi and Verma, 2016; Zhang et al.,
2012). Alternatively, biocathodes have been introduced employing
microorganisms as catalysts that carry out the reduction process by
accepting electrons and protons generated at anode. Biocathodes
encompass certain improvements as their application in the opera­
tional process can generate beneficial substances and assist in
eliminating unnecessary chemical components. Furthermore, bio­
cathodes utilizing algae as catalyst do not necessitate the provision
of oxygen from an external source (Jung and Pandit, 2019). Appli­
cation of microbes in biocathodes as biocatalysts is highly beneficial
for the fact that they are cost-effective, sustainable, display sa­
tisfactory working efficiency at neutral pH and exhibit self-re­
generation potential thus enhancing reduction action at cathode and
A. Nawaz, I. ul Haq, K. Qaisar et al.
enhancing the power generation potential of MFC (Nitisoravut
et al., 2017).
7.1.3. Separating membranes
Protons are generated along with electrons as a result of oxida­
tion reaction taking place at anode and directly influence the power
output of MFC. These protons reach at cathode by passing through a
separating membrane (Rahimnejad et al., 2014). A separating
membrane is sited between anode and cathode electrodes. It is used
for dividing the compartments, proton transport and precludes the
transfer of oxygen from cathode to anode. A proton exchange
membrane suitable for MFC operation should have higher proton
conductivity, tolerance towards chemicals and heat, mechanical
stability, suitable surface area, and low cost (Tharali et al., 2016a,
2016b). A separating membrane can be cation exchange membrane,
anion exchange membrane, salt bridge, bipolar membrane, ultra­
filtration membrane (UFM), microfiltration membrane (MFM), and
ceramic separators. Among cation exchange membranes, Nafion
constitutes the most widely employed membrane for proton ex­
change due to its characteristically high proton and electrical con­
ductivity. Nafion carries negative charges that promote the transfer
of unwanted cations resulting in a drop in pH of anode compart­
ment, disturbing microbial potential at anode. Alternatively, an in­
crease in pH of cathode reduces the cathode efficiency, in turn
affecting the working efficiency of MFC. Additionally, the use of
Nafion is costly. Anion exchange membranes, on the other hand,
increase the efficiency and power output of MFC by facilitating the
transport of protons and prohibiting the transfer of other cations
thereby maintaining the pH gradient (Jung and Pandit, 2019). Other
materials such as glass fiber and glass wool have been utilized as
membrane because of their low cost and positive impact on MFC
performance. Glass fiber exhibits higher efficiency for proton
transport, minimized transfer rate of oxygen, and decreased re­
sistance (Aghababaie et al., 2015).
Owing to greater proton conductivity potential and robustness,
Perfluorosulfonic acid (PFSA)-based polymers referred as Nafion
membranes, have been extensively utilized as proton exchange
membrane in fuel cells. Yet, its reduced resistance to methanol,
decline in proton conductivity at elevated temperature and lesser
humidity settings, influences the effectiveness of Nafion membrane
(RS et al., 2020). The auspicious substitute having greater proton
conductivity and steadiness at higher temperature and humidity are
Sulfonated hydrocarbon polymers such as sulfonated poly (ether
ether ketone)s (SPEEK), sulfonated polyimides (SPI) and sulfonated
poly(ether sulfone) (SPES) (Jiao et al., 2021). Sulfonated polystyrene-
ethylene-butylene-polystyrene (SPSEBS) was also acquired as alter­
nate to Nafion. Similarly, Nanocomposite membranes, due to greater
power generation potential and Coulombic efficiency, greater proton
conductivity, lower cost, and better antifouling characteristics in
contrast to Nafion, have found applications in MFC as PEM (Das et al.,
2018). Among other potential membranes reported are sulfonated
polybenzimidazole (S-OPBI), graphene oxide/SPEEK (GO/SPEEK),
quaternized poly (ether ether ketone) (QPEEK). Thus, suggesting
non-fluorinated separating membranes and those developed with
modification as with nanoparticles as suitable alternatives that can
improve energy output from Microbial fuel cell (Shabani et a, 2020;
Kim and Patel, 2020).
7.2. Effect of pH
During MFC operation, protons produced at anode are trans­
ferred to cathode where they, along with electrons, form water on
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Process Safety and Environmental Protection 161 (2022) 357–373
reacting with oxygen. The rate of proton transport from anode to
cathode influences the pH and the operational efficiency of MFC. The
slow transport rate of proton results in proton buildup at anode
causing anode acidification. At cathode utilization of proton in re­
duction of oxygen and lesser rate of proton transfer to cathode raise
the pH at the cathode. This increase in pH at cathode is associated
with the decline in the reduction of oxygen leading to a decrease in
power generation. (Ivars-Barceló et al., 2018; Samsudeen et al.,
2015). Moreover, maintenance of pH in a suitable range is crucial for
the regulation of microbial metabolism and directly influences the
power generation potential of MFC (Singh et al., 2019). Bacterial cells
display highest metabolic activity at neutral pH conditions. Altera­
tion in pH can lead to changes in physiological parameters including
ionic concentration, biofilm formation, proton motive force and
membrane potential. pH at anode is important in determining the
activity of bacterial cells and affects the electrogenic potential as the
decrease in anodic pH can result in a decline in microbial perfor­
mance (Oliveira et al., 2013). A study aimed at evaluating the effect
of anodic pH on the power output of double chamber microbial fuel
cell employing B. Subtilis strain was carried out. The optimum
growth and activity of B. Subtilis and highest power density of
405 mW/m2 were obtained at anodic pH of 8.6 (Córdova-Bautista
et al., 2017). Another work was conducted using wastewater from
pulp industry and Pseudomonas fluorescens for generation of elec­
tricity assessed the effect of pH on MFC current output. The highest
voltage and current of 0.956 ± 0.009e V and 2.692 ± 0.016e mA
were observed at pH 7 while at pH value above and below than 7
bioelectricity generation potential was reduced (Kaushik and
Jadhav, 2017).
7.3. Temperature
Temperature poses a substantial impact on MFC performance as
different kinetic and thermodynamic properties are governed by this
parameter. Temperature has been found to directly influence the
performance of MFC for bioelectricity production. A rise in tem­
perature results in increased power density output while decreasing
the resistance of MFC (Jatoi et al., 2020).
The presence of microorganisms in operational mechanism of
MFC marks temperature as a highly important factor than in or­
dinary fuel cell. Most of the electricigens are mesophilic micro­
organisms, whereas some psychrophiles, thermophiles and hyper-
thermophilles are also electroactive microorganisms (Butti et al.,
2016). It was reported in a study that microorganisms exhibit op­
timum activity in a temperature range of 30–45 °C temperature and
hence give elevated electric power generation in MFC (Liu et al.,
2011). Variation in temperature from recommended optimum tem­
perature may result in poor biofilm formation and subsequently
reduces MFC potential for bioelectricity generation. Furthermore,
the temperature modifications from optimum temperature can also
result in physical impairment of microbe and cause alterations in
bacterial processes. Different types of bacteria present have different
temperature requirements to develop a biofilm, as soon as the de­
sired temperature is attained the bacterial population will carry out
metabolic functioning accordingly (Jatoi et al., 2020; Song et al.,
2017). Operation within elevated temperature ranges gives improved
voltage output possibly due to increase in microbial growth and
biofilm formation that harbor exalted electrochemical activity (Tee
et al., 2017). A research study investigated the effect of temperature
range 25–45 °C on denitrification of wastewater treatment and
subsequent generation of electricity using dual-chamber MFC. It was
reported that maximum power output for electricity was observed
A. Nawaz, I. ul Haq, K. Qaisar et al.
at 35 °C providing highest coulombic efficiency and current density
of 8.12 ± 0.04% and 6.51 A m− 3 respectively (Wang et al., 2018). A
study investigating the influence of different parameters including
temperature on power productivity employing a dual-chamber MFC
was performed. It was observed that elevation in power density was
achieved on increasing the temperature from 24 °C to 35 °C and the
greatest MFC efficiency was attained at 35 °C (Tremouli et al., 2017).
7.4. Substrate loading rate
Substrate loading rate remains another factor of high significance
that determines the particular flow into the cell of the organic
substrate and the microbial population needed to degrade it.
Enhanced working potential of MFC entails optimizing the substrate
loading rate or organic loading rate as energy output and columbic
efficiency are largely dependent on the rate of substrate utilization
(Abdallah et al., 2019). An increased organic loading rate leads to a
significant rise in power yield and enhanced degradation of sub­
strate. However, elevation in organic loading rate above a particular
limit can cause a drop in energy output with increased degradation
of the substrate. (Goswami and Mishra, 2018). A study evaluated the
impact of different organic loading rates on electricity production
from surgical cotton industry wastewater. A surge in power density
from 42 mW/m2 to 116.03 mW/m2 was noticed as the organic
loading rate was increased from 0.7 gCOD/L d to 1.9 gCOD/L d. This
surge in power output could be ascribed to the efficient consump­
tion of substrate by microbes. Nonetheless, a further increase in
organic loading rate resulted in a decline in power generation
(Tamilarasan et al., 2017).
7.5. Biofilm
Microorganisms constitute the primordial element of MFC for
wastewater treatment and subsequent electricity generation. In
microbial fuel cell, anodic biofilm is of high significance since the
performance of MFC is highly determined by the development and
properties of the biofilm. The bioelectricity production in MFC is
proportional to the development of biofilm at anode. An increase in
biofilm thickness and density promotes greater utilization of sub­
strate as the proportion of bacterial cells that play the role of cata­
lysts, in a thick biofilm is sufficiently elevated (Arbianti et al., 2018;
Choudhury et al., 2017). As the biofilm growth increases at anode it
reduces the polarization resistance of the anode, resultantly in­
crementing the power generation in MFC (Baranitharan et al., 2015).
Certain improvements have been made to enhance biofilm forma­
tion including the utilization of hydrophilic, positively charged
anode as the surface charge of most of the bacteria is negative and
are characteristically hydrophilic (Kalathil et al., 2017). Upgradation
of MFC power generation potential by the application of improved
hybrid biofilm introduced with bacterial multiwalled carbon nano­
tube has been reported. It was noticed that hybrid biofilm displayed
a significant decrease in startup time for MFC operation by 53.8%.
The Hybrid biofilm expressed superior properties of adsorption and
enhanced tolerance to abrupt concentration variations in substrate
than the naturally developed biofilm resulting in an increased cur­
rent density (P. Zhang et al., 2017; L. Zhang et al., 2017). Production of
microbial biofilm having high conductivity, activity, and penetration
power is the core objective of electrode-based microbial modifica­
tion. Genetic modifications of microorganisms for enhancing their
electrogenic potential consequently increments electric output of
MFC utilizing wastewater. Conventional microbial strains are re­
placed by genetically engineered microbes. The selection of suitable
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Process Safety and Environmental Protection 161 (2022) 357–373
genetically manipulated strain for ensuring extended MFC perfor­
mance is crucial. Currently, various methods of molecular biology
including synthetic biology, metagenomics, and microbiomes have
contributed to the efforts made for the production of microbial
biofilm population in MFC (Palanisamy et al., 2019; Angelaalincy
et al., 2018).
Development of the biofilm on the air cathode in a single
chamber microbial fuel cell due to unavailability of separating
membrane and exposure to diffused organic matter exerts detri­
mental impact on the activity of air cathode. Biofilm formation on air
cathode blocks the active sites of catalyst coating on the cathode and
sometimes deleteriously affects the performance of catalysts by its
extracellular substances, consequently affecting the oxygen reduc­
tion potential. In addition, biofilm accumulation at cathode poses
competition to anodic microbial biofilm for substrate utilization. The
growth of aerobic bacteria forming a biofilm obstructs the passage of
protons and charged moieties to the catalyst and prevents the dif­
fusion of hydroxide formed on the cathode disturbing the pH at the
electrode. Aerobic biofilms also exhaust some oxygen thereby af­
fecting reduction reaction at cathode. Various approaches for cir­
cumventing biofouling have been proposed. These methods include
physical methods for electrode cleaning, cleaning by chemical
treatment, application of electric field for removing biofilms, and
modification of electrode surface by integrating catalyst coating with
antimicrobial components (Al Lawati et al., 2019).
7.6. External resistance
External resistance shows a considerable effect on the working of
MFC in terms of electricity generation, COD elimination, and mi­
crobial diversity. An increased external resistance causes a decrease
in current output and decreased external resistance leads to a high
current generation. On operating MFC under decreased external
resistance, reactions at the cathode and electrical activity speed up
because of enhanced transport of electrons to cathode consequently
improving the current output in MFC. Furthermore, low resistance
improves the positive potential of anode resulting in greater energy
gain by microorganisms and increased electrons flux, selectively
promoting electrogenic microbial community (del Campo et al.,
2014). A study elucidating the effect of external resistance on power
generation using a combination of the wood industry and municipal
wastewater was carried out. Maximum current density of 440 mA/
m2 was obtained at 100 Ω that was the lowest of the resistances
investigated (Kloch and Toczyłowska-Mamińska, 2020).
8. MFC based integrated process
Recently, hybrid technologies based on MFC such as Sediment
MFC (SMFC), Membrane-bioreactor MFC (MBR-MFC), constructed-
wetland MFC (CW-MFC), MFC-based denitrification systems,
Desalination MFC (DS-MFC), etc. have been introduced (Xu et al.,
2017). Among these, CW-MFC, SMFC, and DS-MFC have distinct
potential for waste-water treatment and MBR-MFC is competent in
renewable electricity production. However, the major constraint is to
upgrade them on a larger scale and reduce their cost with maximum
output. Therefore, there is a need to conduct research to increase the
stability, performance, and power output of these hybrid technolo­
gies in order to make them economically commercially viable for
their use in the near future. (Nawaz et al., 2020).
A. Nawaz, I. ul Haq, K. Qaisar et al.
9. Future prospects and conclusion
MFCs play a vital role in energy generation and wastewater
treatment as they offer plenty of benefits including environmental
sustainability. However, an optimum design is yet to be invented in
order to reduce extortionate cost and power input. From this review
it can be deduced that double chambered MFCs have an edge over
single chambered MFCs due to high power output and waste water
removal at largescale. However, encouraging results at small scale
are observed with single chambered MFCs as less internal resistance
is required reducing the cost thereby increasing the efficiency. Power
production is enhanced by stacking MFCs into vertical, horizontal,
cylindrical and flat-plate configurations in parallel and series con­
nections. Increasing the reactor size leads to larger volumes conse­
quently reducing power proportionally due to increase in internal
resistance. However, to circumvent this problem, several MFCs can
be connected in a series combination to increase the surface area. In
combination, greater power outputs can be achieved as compared to
individual setups or individual electrodes. So far there is no definite
design to maximize both electricity generation and waste water
treatment simultaneously. Since various pros and cons are asso­
ciated with every configuration and scaling up a singular or com­
binatory design is a challenge. As many factors are yet to be
considered for scalability of MFC in the future e.g. electrode material,
stacking direction, material selection, durability of reactor, and cost-
effectiveness.
Better performance of MFCs is a must for paving the way to an
ideal design, progress can be made by accomplishing a few target
objectives (i) increasing electrogenecity of microorganisms, (ii) ap­
propriate selection of electrode material, (iii)optimizing the opera­
tional factors, (iv) efficient recovery of by-products, (v) minimizing
the capital cost, and (v) maximizing the power output (vi)
Commercializing MFC-based hybrid/integrated technologies.
Microorganisms are crucial in the degradation of substrate and
electron transport from anode. In comparison to pure cultures,
mixed microflora efficiently treat wastewater as they oxidize com­
plex organics. Future research is being conducted by genetically
modifying the microorganisms to increase the electrogenic activity
of microbes. Thereby, enhancing the electron transport and removal
of organic compounds consequently leading to improved MFC per­
formance.
Electrode material acts as an electron acceptor and as an at­
tachment site for biofilm. Selecting an appropriate material of
electrode is very important. Typically, Pt electrodes can be used as
cathode and anodes. However, carbon-based materials like PVC and
graphite have high affinity for microorganisms and are commonly
employed. Further investigation for electrode materials is needed for
long-standing working of MFCs. Operational parameters for example
pH, type of microorganisms, temperature and O2 flow rate are op­
timized in order to get maximum power output and COD removal.
Therefore, the electrode material and the configuration of MFCs di­
rectly impact the cost of the process at large scale. As the capital cost
is increased by the electrode material and operational cost is re­
duced by MFC configuration.
The feasibility of wastewater treatment by MFCs is increased as
high-value byproducts like heavy metals are obtained. Other by-
products like H2 and CH4 can be utilized as energy sources also. Use
of MFCs could be applied in the future for efficient recovery of high
value by-products and reduce the dependence on chemicals for
precipitating these products. Operational cost of MFCs to treat
wastewater is reduced further due to the recovery of heavy metals
that are high-value byproducts. However, capital cost is 30 times
369
Process Safety and Environmental Protection 161 (2022) 357–373
higher than the activated sludge treatment. In order to achieve
sustainable wastewater treatment with bioelectricity production
simultaneously, it is essential to create a balance between the op­
erational and capital cost. Power output can be amplified as the
input power is relatively higher and the output power is not enough
to meet the need of wastewater treatment solely. Additional studies
are required to increase the energy efficiency by storing it in bat­
teries and capacitors thereby cumulating the available energy for
treatment.
Establishing economical technologies is the need of the hour for
sustainable electricity production and degradation of substrate for
wastewater treatment. Developing new technologies or integrating
more than one existing MFC-based technology would exploit by­
products and waste as other energy sources that can be utilized. This
minimizes the carbon footprint, therefore, reduces the load on en­
vironment. Since renewable energy generated from a single tech­
nology is expensive, combing these technologies commercially
would sum up the total renewable energy output thereby making it
economically viable. Substantial research is in progression con­
cerning new materials, configurations, and approaches of MFCs for
scaling up the projects thus filling the gap between the production
and commercialization of bioelectricity generation and wastewater
treatment.
CRediT authorship contribution statement
Ali Nawaz: Conceptualization, Writing-Original draft, Writing
–Review and editing; Ikam ul Haq and Burcu Gunes: Writing
–Review and editing. Kinza Qaiser; Saleha Ibadat Raja; Khola
Mohyuddin, Haseeb Amin: Data curation, Writing-Original draft,
Writing –Review and editing.
Declaration of Competing Interest
The authors declare no conflict of interest.
Acknowledgments
We are grateful to Usama Ikram, graphic designer from
University of Engineering and Technology Lahore Pakistan in helping
us out with figure making to make the review more attractive and
interactive towards the reader.
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