Renewable and Sustainable Energy Reviews 73 (2017) 75–84

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Renewable and Sustainable Energy Reviews

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

Sustainable power generation from bacterio-algal microbial fuel cells MARK

(MFCs): An overview
⁎
Beenish Sabaa,b, , Ann D. Christya, Zhongtang Yuc, Anne C. Cod
a
Department of Food Agricultural and Biological Engineering, The Ohio State University, 590 Woody Hayes Drive, Columbus, OH 43210, USA
b
Department of Environmental Sciences, PMAS Arid Agriculture University, Rawalpindi 46300, Pakistan
c
Department of Animal Sciences, The Ohio State University, 2029 Fyffe Road, Columbus, OH 43210, USA
d
Department of Chemistry and Biochemistry, The Ohio State University, 100 West 18th Avenue, Columbus, OH 43210, USA

A R T I C L E I N F O A BS T RAC T

Keywords: Microbial fuel cells (MFCs) are bioelectrochemical devices that allow the harvesting of electricity generated
Bacteria during anaerobic respiration of selected bacterial species. This technology shows promise in both wastewater
Algae treatment and sustainable bioenergy conversion applications. Bacterial respiration occurs in the anaerobic
Microbial fuel cells anode compartment of the MFC, and is electrochemically coupled with electron acceptors in the MFC's aerobic
Operational and electrical parameters
cathode compartment. This paper summarizes the published results of bacterio-algal MFCs. The use of
microalgae in MFCs has gained interest primarily due to algae's ability to photosynthesize atmospheric CO2,
producing both biomass and oxygen and thereby facilitating the cathodic reaction. These phototrophic
microorganisms can serve as biocatholytes in MFCs because the oxygen produced is an electron acceptor for
the electrons harvested from the anode compartment. The bacterio-algal MFC can provide multiple benefits
including 1) power generation, 2) wastewater treatment, 3) algal biomass cultivation and pigment production,
4) carbon dioxide assimilation, and 5) oxygen production. This review article summarizes not only successful
published results of bacterio-algal fuel cells but also highlights critical operational parameters and their effect on
power generation and output efficiency.

1. Introduction
Among newly developed renewable energy production technologies,
the microbial fuel cell (MFC) is a promising technique because of its
sustainability to directly convert waste into electrical energy [1–4].
MFCs typically consist of two compartments separated by a proton
exchange membrane with an external circuit connecting anode and
cathode electrodes. They can be categorized on the basis of catholyte;
1) abiotic catholyte e.g potassium ferricyanide, nutrient medium etc. 2)
biotic catholyte e.g. algae, cyanobacteria etc. The second category
hereafter is referred as bacterio-algal MFCs and will be the main focus
of the present review. The bacterio-algal MFC is an approach to
enhance the economic and carbon capture characteristics of MFC
technology [5]. In this approach, autotrophic and phototrophic micro-
organisms are employed as a catholyte along with anode-reducing
exoelectrogenic bacteria as an anolyte [5]. Algae can be introduced to
either the anodic or the cathodic compartments of the MFC to serve as
a source of carbon (substrate) or to generate oxygen, respectively. It
can be cultivated within the cathode chamber by passively capturing
CO2 from the atmosphere [7] or artificially sparging more concentrated
CO2 streams into the cathode chamber [8]. The required Sunlight can
be provided naturally or by grow lights under laboratory conditions.
Carbon substrate can be obtained by a variety of media such as glucose,
formate, acetate and dry algae biomass. Direct use of the dry algae as a
substrate is problematic because it is difficult for bacteria to hydrolyse
algal cell walls [9]. Glucose, formate and acetate substrates are simple
sources of carbon which are readily available to bacteria, but they are
more expensive [5,10] as compared to the waste sources such as
domestic wastewater [10], activated sludge [11], landfill leachate [12],
food processing leachate [13], and other waste materials.
The utilization of wastewater as a substrate in bacterio-algal MFCs
for power generation is widely studied inexpensive nutrient source in
laboratory experiments [9,14,15]. Different configurations of bacterio-
algal MFCs with various electrode materials [13,15,16], variety of
substrates and microbial sources have been employed [6,8]. Some of
these techniques have promising results [2,4,9,17] but the environ-
mental and economic life cycle assessment of these systems is needed
for their practical application. In this current review we aim to
summarise the published research on the performance of bacterio-
algal MFCs and their limitations for the commercial applicability.

⁎
Corresponding author at: Department of Food Agricultural and Biological Engineering, The Ohio State University, 590 Woody Hayes Drive, Columbus, OH 43210, USA.
E-mail address: saba.20@osu.edu (B. Saba).

http://dx.doi.org/10.1016/j.rser.2017.01.115
Received 4 May 2016; Received in revised form 25 November 2016; Accepted 17 January 2017
1364-0321/ © 2017 Elsevier Ltd. All rights reserved.
B. Saba et al.
2. Bacterio-algal fuel cell technology
2.1. Algae as biocatholyte
Photosynthesis is one of the most complex biological redox reac-
tions. In this reaction, natural solar energy and carbon dioxide are the
inputs to a process that produces carbohydrates, oxygen and additional
compounds including proteins, pigments, and oils. Autotrophic growth
of algae can be supported in artificial environments. Photobioreactors
[18], algal ponds, and lagoons [19] are some of the common techniques
used to grow and harvest algal biomass. Heterotrophic growth of algae
in the dark, supported by consumption of various carbon substrates, is
a unique ability of microalgae [20]. However, this heterotrophic mode
of algal growth can be subject to contamination and growth competi-
tion from other microorganisms [20]. In photobioreactors, algae
cultivation is operated in mixotrophic mode (i.e., autotrophic+hetero-
trophic). This mixotrophic property of algae makes it a suitable
candidate as an MFC biocatholyte, along with its other abilities such
as oxygen production and CO2 assimilation, and its logarithmic growth
pattern [21]. Bacterio-algal MFCs are operated on the premise that
substrate is oxidized by bacteria in the anolyte and oxygen is produced
by the algae in the catholyte. This oxygen acts as an electron acceptor,
reducing CO2 and producing water and more algal biomass via
photosynthesis [22–24] (Fig. 1). The chemical equations for photo-
synthetic reactions in the cathodic chamber of algal-MFCs are as
follows:
Light reaction
6CO2+12H++12e-→C6H12O6 (biomass)+3O2
Dark reaction
C6H12O6 (biomass)+6O2→6CO2+6H2O
Cathodic reaction
O2+4H++4e- →2H2O
Algal growth in the cathodic chamber is a self sustaining cycle. Over
time, algae growing in suspension can form a biofilm on the electrode
and chamber surface. Algal biofilm directly accepts electrons traversing
the MFC circuit and these electrons penetrate into the algal cell body
[25]. For planktonic algal suspensions, intermediate mediators are
used which can accept electrons from the cathode and deliver them to
the suspended algae. These reduced mediators penetrate algal cells,
deliver their electrons thereby becoming oxidized, and then are
released from the algal cells back into the catholyte [26,27].
Dissolved oxygen within the algal biofilm has been verified to play an
Fig. 1. Concept of mixotrophic biological reactions in algae.
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Renewable and Sustainable Energy Reviews 73 (2017) 75–84
Fig. 2. Schematic presentation of algae-electrode interaction.
important role in the performance of the electrochemical system [28].
Biofilm formation on the cathode acts as a direct mediator in electron
transfer and reduces ohmic and charge transfer resistance losses in the
MFC [29,30] (Fig. 2).
2.2. Bacterio-algal fuel cell configurations
Three major configurations of MFCs are currently in use for
simultaneous growth of bacteria and algae: (1) single chamber MFCs,
(2) two chambered MFCs, and (3) three chambered MFCs. Schematics
of different configurations of MFCs are shown in Fig. 3. Each of these
designs is described more fully below.
Single chamber bacterial-algal MFCs (Fig. 3A), in which bacteria
and algae are grown together in one chamber, are usually configured
with an air cathode [17,31,32]. Single chamber MFCs are membrane-
less, easy to operate, and cost effective in scaling up. Autotrophic and
heterotrophic microorganisms are grown simultaneously so 100%
consumption by algae of the CO2 generated by the bacteria is possible.
In single chamber MFCs, bacterial co-cultures can grow synergistically
with algal co-cultures [17]. Single chambers are easy to manage in lab
as compare to other configurations.
In two chambered MFCs (Fig. 3B), algae and bacteria exist in
separate chambers which are connected by a cation exchange mem-
brane [9,33]. In two chambered MFCs, a light source is usually placed
on algae side. This serves to provide photons for the algal photosyn-
thetic reactions in the cathodic chamber but also illuminates the
bacterial compartment, leading other researchers to cover their anodic
chambers during experiments [10]. In some studies, an external algae
photobioreactor (Fig. 3C) provides the MFC with a continuous supply
of algae [10,34]. Membrane crossover and relatively high internal
resistance are some of the problems associated with dual chamber
MFCs. Photobioreactors provide a continuous source of algae which is
easier to maintain, but the addition of pumping reduces net power
generation. H-shaped (Fig. 3D) two chambered MFCs have also been
used in earlier studies with the disadvantage that the membrane area
between the anolyte and catholyte is very small. Ion exchange between
two chambers is very low, in turn producing low power output.
Membrane fouling and internal resistance can effect efficiency of two
chambered MFCs.
In three chambered MFCs, an additional middle chamber contain-
ing salt water is also present. Cations move towards the cathode, anions
move towards the anode, and partial desalination is observed in the
process (Fig. 3E). The presence of salt water in the middle chamber can
pose a stress on power production and its peak power density has been
observed to be lower than two chambered MFCs [35,36]. Other
configurations of MFCs include a tube shaped MFC which is a kind
of single chamber MFC [31] and a lagoon or algal pond MFC in which
anodic chambers are submerged in a bath of algae within a larger
lagoon. Power production and biomass content can be optimized
according to local weather conditions in any configuration.
B. Saba et al.
Fig. 3. Bacterio-algal microbial fuel cell configurations (A) single chamber (B) Dual chamber (C) dual chamber with photobioreactor (D) three chamber with desalination (E) H type
dual chamber.
2.3. Types of innocula in bacterio-algal MFCs
Common sources of innocula for MFC anodic chambers include
pure microbial cultures, defined microbial consortia, and undefined
microbial consortia such as municipal wastewater, anaerobic bioreac-
tor effluent, anaerobic sludge [15,31,33]. Synthetic wastewater [37]
and rumen micro-organisms [38] have also been used. Rumen micro-
organisms have added benefits such as faster acclimatization as
compared to municipal wastewater microorganisms and the ability to
hydrolyse cellulosic biomass substrates. Leachate from landfills and
food waste has also been successfully used [39]. Some single specie
inoculation in the anodic chamber have been used including Geobacter
sulferreducens [17], and Shewanella oneidensis [40]. Biofilm and
suspended planktonic analysis of the microbial community in the
anode chamber of a MFC inoculated with activated sludge showed
Desulfovibrio sp., P. aeruginosa, Cytophaga xylanolytica,
Dechloromonas sp., Thiomonas perometabolis, and Cytophaga sp. as
dominant species [9].
The source of innocula in the cathodic chamber of bacterio-algal
MFCs is algae. Algae are introduced to MFCs to increase the oxygen
concentration in the catholyte and to absorb CO2 either from ambient
air or diverted from the anodic chamber. Chlorella vulgaris [6,14,18] is
a common cathodic algae. Other species of Chlorella [40], marine algae
[33], blue green algae and cyanobacteria [31] have also proven their
usefulness in bacterio-algal MFCs. Table 1 summarizes algae used in
bacterio-algal MFCs as documented in the research literature. A variety
of algal species including pure culture, co-cultures, and mixed cultures
have been employed.
2.4. Anolytes in bacterio-algal MFCs
Electricity production and COD removal characteristics of MFCs
depend primarily on the redox reactions in the anode chamber.
Different carbon sources have been studied for use in the anolytes of
bacterio-algal MFCs. Most common are glucose [14], formate and
acetae [17]. Other prepared sources include LB medium [40], GSMM
[33], Scenedesmus algae in powder form [8], GM medium [29], fruit
industry liquid waste [23], and synthetic wastewater [37]. MFC
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Renewable and Sustainable Energy Reviews 73 (2017) 75–84
membrane type and internal resistance are the factors effecting power
generation. Columbic efficiency and power production depends on
number of factors like resistance of membrane, high ion generation in
anodic chamber, oxygen cross over through membrane or membrane is
highly porous. Therefore low resistance and high conductivity and non
porous, highly selective membranes should be employed. Membrane
fouling, high COD and low pH are some other factors which can lower
membrane efficiency [41]. To solve these problems, membrane pre-
treatment and continuous monitoring of the anodic chamber’s internal
conditions are necessary. Cation proton exchange membranes are used
in MFCs depending upon reaction kinetics. Single chamber MFCs are
membrane-less, which avoids membrane losses but can cause oxygen
to more easily cross into the anaerobic anode chamber. Depending on
the type of micro-organisms present in the anolyte, this can cause
facultative anaerobes to switch metabolic pathways away from elec-
tricity generating anaerobic respiration, can be deadly to obligate
anaerobes, and can cause changes in microbial consortia for mixed
cultures. For example, aerobic microorganisms present in wastewater
sludge can outcompete anaerobic microbes leading to lower power
production. Table 1 summarizes the type of membranes used in dual
chamber bacterio-algal MFCs.
2.5. Catholytes in bacterio-algal MFCs
The major difference between common MFCs and bacterio-algal
MFCs is the presence of algae in cathodic chamber. Algae oxygenates
the cathodic chamber and yields additional algal biomass and pig-
ments. Oxygen generated by algal photosynthesis accepts electrons at
the cathode. Algae is tolerant of changes in cation concentrations due
to other ions crossing the membrane from the anode chamber. Algal
biomass produced can be used as a valuable by-product either as a
source of animal feed or for energy and bioproduct generation via
anaerobic digestion [42] or other bioprocess. Biomass production in
batch reactions with higher hydraulic retention times (HRT) produces
more algae than a continuous system with lower HRTs. By circulating
algal biomass between a separate photobioreactor and the MFC’s
cathodic chamber [34], this disadvantage of continuous operation
can be overcome.
B. Saba et al. Renewable and Sustainable Energy Reviews 73 (2017) 75–84

Table 1
Power density of Bacterio-algal MFCs.

Cathodic contents Anodic contents Power output (mW/m2) References

1 Blue green algae Anaerobic wastewater sludge 114 [31]

2 Chlorella vulgaris Activated sludge 13.5 [11]
3 Scenedismus obliquus GM media only 153 [29]
4 Desmodesmus sp. Synthetic wastewater 99 [37]
5 Chlorella and blue green Phormidium wastewater 263 [10]
6 Chlorella vulgaris Enriched bacterial consortium 62.7 [16]
7 Mix algal culture Municipal wastewater 11.5 [15]
8 Laminaria saccharina liquor (autoclaved) Wastewater microbial consortium 218 [33]
9 Laminaria saccharina liquor Wastewater microbial consortium 118 [33]
(Microwaved)
10 Chlamydomonas reinhardtii Geobacter sulferreducens 41 [17]
11 Chlamydomonas reinhardtii Geobacter sulferreducens +Acetate 630 [17]
12 Chlamydomonas reinhardtii Geobacter sulferreducens +Formate 140 [17]
13 Mix algae Activated anaerobic sludge – [34]
14 Spirulina platensis – 10 [25]
15 Scenedismus obliquus Activated sludge 1.78 W/m2 [9]
16 Chlorella vulgaris Scenedesmus enriched anaerobic activated sludge 514 [18]
17 Chlorella vulgaris Domestic wastewater 5.2 W/m3 [14]
18 Chlorella vulgaris – 48 mA/m2 [6]
19 Mixed algal culture Anerobic suspension from growth reactor 57 [17]
20 Synechococcus leopoliensis, Anabaena cylindrical, Chlorella pyrenoidosa Shewanella oneidensis 0.0033 [40]

Table 2 successful growth of Chlorella vulgaris. The different varieties of algal

COD removal efficiency of bacterio-algal MFCs.
S.No. COD removal (%)
1 81
2 80
3 80
4 90
5 88
6 74% in cathodic chamber and 37% in anodic chamber
during spring
7 58% in cathodic chamber and 27% in anodic chamber
during summer
A limiting step in power generation is the rate of oxygen reduction
to water in the cathode chamber. Oxygen reduction can be enhanced by
applying a catalyst at the cathode [43] or by adding a strong reducing
salt such as potassium ferricyanide [44] to the catholyte; other methods
for increasing oxygen availability include sparging air or pure oxygen
into the cathode chamber or by rotating the electrode [45]. However,
algae cultivation within the cathode chamber can provide a more
consistent and continuous source of oxygen [11].
Thick algal biofilms on the cathode surface can limit oxygen
diffusion and MFC performance [46]. Algal biofilm growth and
thickness depends on the electrode’s surface texture; highly porous
and/or rough surfaces favour thick algae biofilm growth while smooth
surfaces support thin biofilm growth which is the better option [16].
Monitoring algal growth by collecting optical density measurements in
the cathode chamber is challenging given that the growth occurs in
three forms: as algae biofilm on the electrode and chamber walls, as
suspended algal aggregates, and as suspended biomass in solution.
Algae biomass monitoring by weighing dry biomass samples or
assaying for protein content are other feasible options.
A wide variety of catholytes has been used in bacterio-algal MFCs
including Bold's Basal Medium with 300 mg/dm3 algal biomass [6,11],
a mixture of 1 g NH4Cl and 0.13 g of KH2PO4_H2O in tap water [10].
Gouveia et al. [47] produced 2800 mg/l of biomass using 50 mM PBS
as the catholyte nutrient medium. Bold Basal Medium containing algae
(0.93 g/l), sodium bicarbonate (NaHCO3, 2 g/l) as a carbon source, and
100 mM PBS was the catholyte composition reported by Kakarla and
Min [29]. Mohan et al. [48] used domestic sewage as their catholyte
with the addition of glucose and urea as carbon and nitrogen sources
respectively, while Wang et al. [14] used 50 mM NBS with BG11 for
species across all of these studies required different growth media and
exhibited different growth parameters which in turn caused variation in
References
power production.
[10]
[31]
[11] 2.6. Electrode materials
[18]
[33] Different electrode materials have been studied for use in bacterio-
[48] algal MFCs. In search of improved electrode performance, researchers
[48]
have studied a variety of materials, surface geometry area enhance-
ments, and surface modifications. Carbon based electrodes are used
due to their chemical stability, electrical conductivity, low cost, and
support of bacterial adhesion to form stable biofilms. Commonly used
materials are graphite plates and rods, carbon fiber brushes, carbon
cloth, carbon paper, carbon felt, carbon nanotubes, and granulated
graphite. Among these, plain graphite and graphite felt are the most
studied forms of cathodic material in bacterio-algal MFCs [17]. Various
coatings have been explored including platinum and 10% Teflon and
coated carbon paper has also been reported to support algal biofilm
formation on the cathode. Table 3 summarizes the types of anodes and
materials reported for use in bacterio-algal MFCs. Most of the studies
used the same materials for both anode and cathode, but in some
studies they have used different materials to increase surface area for
biofilm formation on one or the other electrode [8]. Table 4 sum-
marizes the use of different electrode materials in published studies,
their drawbacks, and benefits. Materials for MFC chamber construction
ranges from a variety of plastics to glass. Use of any specific material
has not proven to have any effect on MFC power production.
Most frequently, copper wire was used for electrode connections
[13,29,54], but some researchers have used titanium wire [9]. The
advantages of copper wire include its high thermal and electrical
conductivities, and its mechanical characteristics of high tensile
strength and ductility. The corrosion resistance of copper also makes
it very suitable for use in liquid MFC systems [49]. However price of
copper wires increases the capital cost of MFCs.
3. Operational parameters
3.1. Light intensities
In the algal cathodic chamber both oxidation and reduction
reactions occur when algae are provided 12 h light and 12 h dark

78
 B. Saba et al.

Table 3
Types of electrodes and configuration of bacterio-algal MFCs.

Cathode Anode MFC material Membrane Light intensity Comments Reference

Two chamber
1 Graphite carbon Acrylic PEM 6–12 W Different light intensities were evaluated in MFC with algae growth and low light intensity is producing high [10]
12–18 W power output is proved in this study.
2 Carbon cloth – PEM 11 W Light was provided in 12 h day night cycles, CO2 was bubbled through the system and lagooning microbial fuel [15]
cell was studied in this work.
3 Graphite – PEM 96 µ E/m2S Pigment extraction and quantification was performed under normal and starved nitrogen conditions, Chlorophyll, [16]
lutein/zeuxathin, βcarotene and cathaxathine were among the main extracted pigments.
4 Plain graphite felt Plexi-glass CEM 3000 lux Light intensity MFC output has a linear relationship of increase in illumination negatively effects cathodic [37]
resistance
5 Toray carbon cloth 10% Teflon PEM 11 W CO2 bubbling for 30 min is enough time to get system working and high polarization resistance in algal cathode [11]
was observed than anode.
7 Graphite felt – – – Algae liquor was added in anodic chamber as co-substrate for microbes and power production capacity was [33]
compared with glucose as carbon source
8 Carbon fiber veil – CEM – They used algal photoreactor attached with cathodic chamber and catholyte was continuously circulated between [34]
chamber and photoreactor
9 Carbon cloth Carbon fiber brush acrylic CEM 2000 lux CO2 produced in anode is diverted cathodic chamber for algal growth and very reduced algae growth was observed [37]

79
when fed with acetate
10 Carbon brush Carbon cloth+Pt acrylic CEM – Pre-treated (sonicated) and lipid extracted algae produces less power as compare to non pre-treated and non- [9]
catalyst extracted algae.
11 Carbon cloth Carbon fiber brush Plexiglas CEM 25 W Open cathodic chamber are helpful in ambient CO2 sequestration with simultaneous voltage output without [14]
aeration.
12 Graphite plates Poly acrylate PEM – Chlorophyll content produced by algae is higher in spring while lower in summer while VFAs are higher in [48]
summer and lower in spring.
13 Carbon fiber veil Acrylic CEM 30 W Oxygenic phototrophic biofilms can enhance cathodic performance [40]
Single chamber
14 Graphite carbon Gliding gold mesh Glass Membrane less – [25]
15 Carbon cloth Graphite felt roll PVC tube Membrane less – 89% COD removal, 96% NH4 removal along with > 90% microsystein (neurotoxins) removal was observed in this [31]
study
16 Carbon paper Platinum Plain carbon paper Glass bottle Membrane less 5000 lumen Algae producing oxygen in electrode chamber produces 32% higher power output than mechanical aeration [29]
coated
17 Air cathode Graphite felt – Membrane less 20 W/m2 Light to electricity conversion is result of syntrophy between phototrophs and autotrophs but oxygen production [17]
by phototrophs is negatively effecting anaerobic G.sulferreducense in single chamber
2
18 Graphite felt – Membraneless 81 µmol/m s Higher algal biomass can be obtained at lower resistance When wires are connected for current production CO2 [6]
generation increases and CH4 generation decreases.
Renewable and Sustainable Energy Reviews 73 (2017) 75–84
B. Saba et al.
Table 4
Listed are some types of electrode materials with advantages and drawbacks of materials used in MFCs.
Electrode types Benefits
1 Graphite plate Defined surface area, high conductivity, inexpensive
2 Graphite fiber brush High porosity 95–98%
3 Carbon cloth Large porosity
4 Carbon paper Easy electrical connection
5 Carbon felt Easy electrical connection
6 Carbon nanotubes Very high surface area
7 Graphene modified Better electrochemical performance,
Anti-corrosive
cycles. During light period photosynthesis, algal biomass production
and oxygen production (oxidation) occurs, and during the dark period
algae consumes oxygen and oxidizes organic matter (reduction) pre-
viously produced. Algae produces 60% of light cycle electricity during
dark cycle, and substances other than oxygen act as electron acceptors
[11]. A recent study [37] suggested that higher light intensity and
optimal dark and light cycles decreased cathodic resistance and
increased power production, but Juang et al. [10] observed that light
power had a inverse relationship with current density, voltage produc-
tion and columbic efficiency. Juang et al. [10] applied two different sets
of lights, one rated at 6–12 W and the other 12–18 W; higher power
production was observed at the lower light power. Relatively higher
power was observed when the anodic chamber was covered; one reason
may be that some algae present in anodic chamber start growing in the
presence of light and produces oxygen. The difference between power
production with covered versus uncovered anodic chambers was
4.41 mW/Kg COD/day. Wu et al. [37] observed 1.5 times higher power
production when light intensity was raised from 1500 lx to 3500 lx. The
profound effect of increasing light intensity on power production is
evident from these studies.
3.2. Carbon dioxide
To promote algal growth, different media and nutrients were
supplemented, along with sparging of CO2 for enhanced photosynth-
esis. Campo et al. [11] found that 30 min sparging of CO2 into the
cathodic chamber produced optimal growth while Wang et al. [14] and
Cui et al. [8] diverted CO2 produced by bacteria in the anodic chamber
into the cathodic chamber; their studies suggested that continuous
bubbling of CO2 was not required. Daytime electricity production
strongly depends on organic loading rate and light irradiation.
Myerse et al. [50] demonstrated that algal cells prefer to use CO2 in
the presence of light and organic carbon. Cao et al. [51] observed
inorganic carbon (IC) in the cathodic and anodic chamber and came to
conclusion that algae fix CO2 by utilizing IC and additional bacterial
CO2 emitted from anodic chamber which can also permeate through
the membrane and be consumed by microalgae. Solubility of CO2 poses
a negative effect on algae growth in early stages. CO2 interaction with
water forms H2CO3 and lowers pH [52]. To overcome this pH problem,
algal inoculation should be initially high [53]. CO2 concentration also
affects the lipid content of microalgae. The cells produce poly-unsatu-
rated fatty acids under high CO2 concentrations [54]. A 6% lipid
content increase was observed accompanied by a 10–15% increase in
CO2 supply [55]. Addition of carbon dioxide does not improve biodiesel
quality however biomass production rate and quantity of biodiesel
produces is effected positively [56].
3.3. Dissolved oxygen
Dissolved oxygen (DO) concentration in the catholyte is one of the
most important parameters which control MFC performance [57]. DO
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Renewable and Sustainable Energy Reviews 73 (2017) 75–84
Drawbacks References
Low surface area [83]
Good bio-compability
Clogging, un-excess able sites for bacteria [84]
relatively expensive [77], [85]
Non durable, high resistance, fragile [86]
High resistance, [86]
clogging
Toxic to microbes [85], [87]
Expensive, [88]
highly brittle
concentration fluctuates diurnally and is also affected by temperature.
During night time hours DO levels drop due to algal respiration and
inhibition of photosynthetic activities, while in daylight hours it is
raised as a product of photosynthesis. Jung et al. [10] observed that
when temperature was raised from 27 to 31.2 °C, DO concentration in
catholyte containing Chlorella vulgaris raised from 3.7 to 6.8 mg/L. In
comparison, the solubility of oxygen in fresh water is reduced from
7.97 to 7.43 mg/L when temperature increases from 27 to 31 °C
(Oxygen Solubility Table [updated 2016]). Temperature effects on
DO in an algal enriched catholyte may be due to a combination of
responses to changes in light intensity of the artificial lamp as well as
ambient temperature variations. When the power of the artificial lamp
is increased from 6 W to 26 W, the rate of photosynthesis and DO
concentrations are both raised [10].
Temperature, DO, and rate of photosynthesis are all interdepen-
dent. Critical values of DO for maximum MFC power production ranges
between 4.5 and 5.5 mg/L [10,58]. Wu et al. [37] observed 13.2 mg/L
DO at higher illuminance (3500 lux) which increased photosynthesises
and thus algal DO production. Supply of more light reduces the power
production of MFC. As light levels increase, the resulting higher oxygen
gradient between the MFC chambers also may drive oxygen to cross
over from the cathodic chamber to anodic chamber. Too high of light
intensity can actually slow down the photosynthetic process, called
photo-inhibition [14]. Lobato et al. [15] observed that cathodic
processes, and specifically DO concentrations, control the overall
performance of the bacterio-algal MFC. For the two selected light
intensities (26 µE/m2s and 96 µE/m2s) reported by Gouveia et al. [47],
the higher light intensity resulted in 10xhigher growth rate of micro-
algae and 6 x higher power production, along with increases in oxygen
concentrations and pigment production. Bio cathodes allows expensive
noble catalysts employed for the oxygen reduction can be replaced by
algal biocathodes [24,34].
4. Electrochemical parameters
4.1. Polarization curves
Changing external loads lead to alterations in the power generation
profile of the MFC. Polarization curves are usually plotted as power
density (PD) and corresponding voltage versus current density under
varying external resistances. The external load at which maximum
power density is obtained during polarization testing is the cell design
point (CDP). In most MFC research, the external resistance step test
procedure which is used to generate polarization curves is applied
when the system's performance is stable which produces a constant
output over the cycles. Other bacterial-algal MFC researchers prefer to
perform step tests with every light-dark cycle of the photosynthesis
process resulting in the generation of several polarization curves; for
example Chandra et al. [59] reported polarization curves for their
bacterio-algal MFCs with 6 cycles of day and night performance. Their
observations showed improvement in power density from 17 mW/m2
B. Saba et al.
(cycle 1) to 72 mW/m2 (cycle 6). The reason is that over time the total
biomass has increased, resulting in more electron transfer from
biocatalyst to anode. Over progressive cycles, ohmic losses (due to
losses from electrodes, electrolytes, and membranes) and activation
losses (due to biomass growth and metabolism kinetics) were observed
to decrease.
Power output of an MFC can be maximized when external
resistance is equal to internal resistance. Higher power production
depends on internal resistances, and in a bacterio-algal MFCs internal
resistance is not affected by changes in light intensity [60].
4.2. Cyclic voltammetry
Cyclic voltammetry (CV) is used to evaluate the surface-controlled
electrochemical processes that represent the redox behavior of electro-
chemically active microorganisms. CV applies varying potential differ-
ences across two electrodes, a working electrode and a Ag/AgCl
reference electrode, allowing quantification of the electrical current
generated as a result of redox reactions within the tested system which
can be analysed by plotting a voltammogram [61].
In studying the behavior of bacterial-algal MFCs, Yu et al. [31]
observed an oxidation peak at +0.14V and a reduction peak at -0.30V
(vs. an Ag/AgCl reference electrode) and discovered that blue green
algae could catalyze electron reduction at the MFC's cathode. The study
also revealed proportionality in the anodic and cathodic peak currents
at varying scan rates which suggested that the redox reaction was
repeatable over time. Chandra et al. [59] observed marked improve-
ment in electrochemical behavior after every feeding event in their
bacterial-algal MFCs which may have been due to incremental growth
in total biomass and adaptation of both anodic bacteria and cathodic
algae to the system's micro-environment. Increases in oxidation peak
values after fresh substrate loading was also reported by Mohan et al.
[48]. Mixing produced by mechanical agitation of the catholyte solution
also enhanced cathodic peak potential [29] causing voltage and current
to increase by 26 mV and 4.6 mA respectively.
4.3. Electrical impedance spectroscopy
Electrical impedance spectroscopy is a technique to determine the
impedence response of an electrochemical system by applying an
alternating circuit (AC) potential and recording the output current
signal which is graphed on a Nyquist diagram, a polar plot representing
a system's frequency response. Impedence is a complex combination of
resistance, capacitance, and inductance effects. To determine ohmic
and polarization resistance values for the studied system, a suite of
standard electrical circuits are evaluated for goodness of fit to
experimentally obtained impedance data. The selected equivalent
circuit model consists of an ohmic resistance component (representing
the MFC membrane, biofilm, and solution resistances) and polarization
resistance (representing charge transfer between anode and cathode).
High polarization resistance is indicated on an Nyquist diagram by an
unclosed semicircle plot when analysing a complete cell (anode and
cathode) and a closed semicircle plot when analysing the anodic side of
the cell only.
The first intersection of Nyquist plot with the x-axis represents the
system's ohmic resistance, and the projected point of the intersection
between the curve and the x-axis represents total impedance due to the
membrane, solution, and charge transfer effects [62]. Campo [11]
observed polarization resistances of 145000 Ω for a complete cell and
1069 Ω for the anode compartment only on the tenth day of operation.
The results clearly indicate very high polarization resistance of the
cathodic compartment as compared to the anodic polarization resis-
tance. High polarization resistance may be due to absence of catalyst.
In bacterio-algal MFCs, algae produce oxygen which accepts electrons,
but the algae do not act as bioelectrical catalysts. Manohar et al. [63]
also observed high cathodic polarization resistance using algae in the
81
Renewable and Sustainable Energy Reviews 73 (2017) 75–84
catholyte. Ohmic and polarization resistances decreased over time [29]
which could be due to excretion of organic substances from growing
algal cultures, such as amino acids, hormones and lipids [64] but more
studies are needed.
5. Perspective benefits of bacterio-algal MFCs
5.1. Useful algal by-products
Evaluation of the pigment content profile of algae is another way to
validate alga growth and photosynthetic activity. Chlorophyll content
reflects the photosynthetic activity of algae in the cathode chamber for
simultaneous energy supply, oxygen release, and CO2 utilization.
Mohan et al. [48] observed 0.8 g/L algal biomass growth in the spring
and 0.9 g/L in the summer. Chlorophyll content increased from
7.21 lg/mg to 15.61 lg/mg with operating time. Seasonal changes in
terms of temperature change affects algal growth and its associated
photosynthetic activity. Elevated summer temperatures can result in
suppressed chlorophyll content. Higher temperatures also lower dis-
solved oxygen solubility. Both adversely affect microbial fuel cell
performance by lowering the amount of oxygen available for as
terminal electron acceptors in the catholyte.
Gouveia et al. [47] observed 0.8% (w/w) (g pigment/100 g dry algae
biomass) as compared to 0.6% (g pigment/100 g dry algae biomass) at
higher light intensity (96 µE/m2s). Pigment content fractionation by
chromatographic techniques can confirm the presence of lutein,
canthaxanthin, and β-carotene. Identified chlorophyll types included
chlorophyll a, b and pheophytin and pheophorbide as degradation
products. Higher light intensities induced the carotenogenesis process
characterized by production of β-carotene and secondary carotenoids
[47,65]. Micro-algal biomass production can be accelerated under light
and nutrient (nitrogen) stress [65]. The identified carotenoids (β-
carotene and secondary carotenoids and lutein) have added value as
antioxidants, pharmaceuticals and nutraceuticals [66]. Stimulating
micro-algal growth under stress conditions can give rise to valuable
compounds without compromising power output [47].
Use of blue green algae in MFCs not only produces pigments but
also releases potent toxins. Micocystins are the toxins produced by blue
green algae in lake waters when the algal cells rupture and die [67,68].
Micocystin cannot be treated with conventional treatment technologies
such as coagulation, flocculation, sedimentation, and filtration [69,70].
Presence of blue green algae in bacterio-algal MFCs cannot effectively
harvest bioelectricity but can successfully treats toxic micocystins
Micocystin-RR and –LR types (90.7% and 91%) [31]. Single chamber
MFC anode provides a more favourable pathway of Micocystin
decomposition under anoxic condition [71]. Synthetic plastic produc-
tion from algal polysaccharides is another useful product of algal
biomass. Cyanobacteria is an excellent feed stock for bio-plastic
production due to its high yield and ability to grow in variety of
environments [72,73]. Tuccar and Aydin, [74] evaluated micro algal
biodiesel in diesel engines and compared with diesel values. The results
showed improved emission values however reduction in torque and
brake power was observed. Mehrabadi, et al. [56] compared biodiesel
quality produced from algal fatty acid methyl ester with pure diesel and
concluded that biodiesel from algal fatty acid is of low quality and
cannot be used directly as transportation fuel.
5.2. Power production
Pre-acclimated microorganisms in lab scale bioreactors or microbes
freshly obtained from natural sources can be employed in bacterio-
algal MFCs. The power output of MFCs strongly depends on low
resistance in the external circuit, high permeability ion exchange
membranes [41], reducing conditions, and high oxygen concentrations
in the cathode chamber [2,15]. Table 1 summarizes power production
potentials reported for different bacterio-algal MFCs which ranged
B. Saba et al.
from 0.0033 mW/m3 to 5.2 W/m3 in maximum power output although
reported units varied. Recent studies showed relatively lower power
generation in bacterio-algal MFCs but these studies were performed to
optimize other parameters such as effect of light intensity, seasonal
effect, pigment production potential, and effect of separating the photo-
bioreactor from the MFCs. Light intensity and duration [10], tempera-
ture [48], CO2 supply [11] are some of the parameters which strongly
affect power generation and MFC efficiency. Algal biomass has been
used in cathodic chamber (as phototroph and electron acceptor) and in
anodic chamber (as an electron donor/substrate). Chandra et al. [59]
observed 165 ± 10 mV (cycle 1) to 535 ± 10 mV (cycle 6) open circuit
voltage (OCV) in a single chamber bacterio-algal MFC. Improvement in
overall output was correlated with total biomass growth observed
0.55 g/L (cycle 1) to 3.24 g/L (cycle6). Power production also varied
with light and dark phases of algal growth. A 350 mV drop in OCV was
observed in the dark phase of algal growth [16]. Wu et al. [37] observed
99 mW/m2 power density at a current density of 380 mA/m2 employ-
ing Chlorella vulgaris in the cathodic chamber. However different algal
strains behave differently [75]. Algae assisted cathodes can generate up
to 1.3 times higher maximum power than non-algae assisted cathodes
[76]. Variations in algal concentration did not affect OCVs 805 ± 12 mV
and open circuit potentials (OCPs), anode (−537 ± 4 vs saturated
calomel electrode (SCE)) and cathode (260 ± 7 vs SCE). Maximum
power production from bacterio-algal MFCs varies with algal species,
growth conditions and substrates, however research has proved that
this type of MFC is capable of successful power production.
5.3. Wastewater treatment
Present wastewater treatment technologies are heavy users of
energy, but MFCs can serve as an energy-generating wastewater pre-
treatment systems [14]. Wastewater can provide ample supply of
nutrients, support large capacity of biofuel production and can be
integrated with existing wastewater treatment infrastructure.
Reduction in Chemical oxygen demand (COD) is one of the ways to
measure wastewater treatment efficiency. The anaerobic consortia in
an MFC's anodic chamber can easily biodegrade organic substrates and
thus reduce COD. In several MFC studies, up to 90% percent COD
removal was observed [8,33,48]. Table 2 summarizes reported COD
removal efficiencies of bacterio-algal MFCs in the literature. Juan et al.
[10] observed 80% COD removal while employing microorganisms
originally collected from an oxidation ditch and supplying them with
982 mg/l and 1266 mg/L of synthetic wastewater. Gadhamshetty et al.
[33] and Campo et al. [8] also observed more than 80% COD removal
using wastewater microbial consortia in MFC. Mohan et al. [48]
observed 74% COD removal in the anodic chamber and 58% COD
removal in the cathodic chamber. Relatively few studies have studied
COD removal in bacterio-algal MFCs, but it is a promising aspect of the
technology that can enhance the significance and practical utilization of
bacterio-algal MFC technology on a commercial scale. However, the
COD removal process can be hindered by several limiting factors,
including electron consumption by methanogenesis, aerobic respira-
tion by the cathodic biofilm, and oxygen crossover [77].
Traditional activated sludge systems creates additional indirect
environmental burdens in the form of sludge disposal which costs
50% of WWT operational cost. Biological nutrient removal also needs
addition of tertiary treatment in WWT plant while using algae
Selvaratnam et al. [78] achieved reduction in nutrient levels to
regulatory discharge level. Thermophilic and acidophilic algal strains
can produce double the biomass per unit nutrient intake than bacterial
system and generation of 20% more net energy. 1 kg of BOD removal in
algal system can generate enough biomass that can anaerobically
digested to generate methane equivalent to 1 kW h of electric power
[79].
Wastewater-algae life cycle assessment studies of Clarens et al. [80]
compared environmental impacts of bioenergy from algae and other
82
Renewable and Sustainable Energy Reviews 73 (2017) 75–84
traditional crops including corn, canola and switchgrass and significant
environmental benefits of using wastewater algae cultivation were
found. Environmental and economic performance of micro-algal
biofuels can not compared with traditional fossil fuels without the
replacement of energy-intensive and expensive fertilizers [81,82].
6. Limitations and summary
Bacterio-algal MFC has proven to be successful at simultaneous
electricity generation and wastewater treatment. Despite the potential
benefits, bacterio-algal MFCs have some limitations and barriers to
successful commercialization. These MFCs are affected by seasonal
changes; for example Mohan et al. [48] reported observing low activity
during the summer season as compared to winter. The optimal design
for a bacterio-algal MFC has not yet been achieved. More detailed
studies of symbiosis between the different species of microorganisms in
bacterio-algal cultures are needed. This paper summarizes the most
recent published research in the field of bacterio-algal MFCs, present-
ing power generation data along with reported sources of substrates,
catholytes, and anolytes.
Future developments of bacterio-algal MFCs in closed systems can
provide 24 hour energy production without current limitations of
catholyte replacement in non-algal MFC systems. Bacterio-algal
MFCs have additional benefit of producing algal biomass which can
be transformed into biofuels and other bio-products via anaerobic
digestion, fermentation, or trans-esterification [89]. However to pro-
duce biomaterials and biodiesel from algal biomass needs pre-treat-
ments and complex extraction methods which increases their price and
hinders scaling up of the systems [90]. Use of algae in the anode
chamber or as substrate, lowers level of out put while in the case of
microalgae-assisted cathodes, the results are promising [91]. To
maximize microalgae power generation much research needs to be
done.
Following recommendations are summarized based on the review.
1. Selection of right type of algal strain or bioengineered strain to
maximize lipid production in bacterio-algal MFCs is required
2. Optimization of technological development in MFC power genera-
tion for the right kind of configuration needs more research
3. Type of biofuel and bio-oil extraction process needs to optimized for
economical bio-oil production
4. Life cycle analysis of optimized MFC is required to suggest this
technology on commercial scale
5. Economic life cycle assessment and optimization of MFC construc-
tion materials to propose bacterio-algal MFCs an economical
technology is required
6. Integrated systems should be promoted to propose solutions of
different problems such as wastewater treatment, nutrient removal,
biomass pre-treatment etc.
Acknowledgements
Beenish Saba gratefully acknowledges a Predoctoral Fellowship
from the Fulbright Program. Grant ID 17130788.
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