ChemTexts (2019) 5:4

https://doi.org/10.1007/s40828-019-0078-3

LECTURE TEXT

Bed electrodes in microbial electrochemistry: setup, operation

and characterization
Jose Rodrigo Quejigo1 · Sara Tejedor‑Sanz2 · Abraham Esteve‑Núñez3,4 · Falk Harnisch1 

Received: 30 November 2018 / Accepted: 3 January 2019

© Springer Nature Switzerland AG 2019

Abstract
Microbial electrochemical technologies have become a vital field of interest in the last two decades. Their reactors are of
interest for a large community of scientists working in environmental engineering and technology, biochemistry, electrochem-
istry, physics, mathematical modeling, microbiology and other disciplines. Due to the fascinating fundamentals and the high
promises for application at the horizon, the field is increasingly reflected in the curricula of students of the aforementioned
disciplines. The main motivation for this article is to give scientists and students an overview on one specific sub-topic: bed
electrodes and bed electrode reactors. After a brief introduction, these granulated electrodes are analyzed from an engineer-
ing, electrochemical and microbiological point of view. Thereby we guide the potential future operator for deciding which
biotechnological processes and applications under which operational conditions (i.e. fixed-bed electrodes versus fluidized
bed electrodes) may benefit by using bed electrodes. Special focus is given to the electrochemical and microbial characteriza-
tion of granules. Thus we discuss a recent tool that opens the possibility to survey the electrochemical behavior of microbial
biofilms on bed electrodes—the e-Clamp. Finally, two case studies of bed electrode reactors are briefly discussed.

Keywords  Bed electrode · Microbial electrochemistry · Microbial electrochemical technologies · Fluidized bed electrode ·
Extracellular electron transfer

Introduction microbial broth. Yet, it took until the dawn of twenty-first

The capacity of living microorganisms to couple their
metabolism to electrodes is known since the beginning of
the twentieth century [1]. The electrons derived from the
oxidation of organic and inorganic compounds (often termed
“substrate”) are transported and transferred to the electrode.
This electrode is then serving as the terminal electron accep-
tor (TEA). In early experiments artificial, i.e., exogenous,
compounds being usually redox dyes were added to the
* Falk Harnisch
falk.harnisch@ufz.de
1
Department of Environmental Microbiology, Helmholtz
Center for Environmental Research GmbH-UFZ,
Permoserstraße 15, 04318 Leipzig, Germany
2
The Molecular Foundry, Lawrence Berkeley National
Laboratory, 1 Cyclotron Road, MS 67‑5113, Berkeley,
CA 94720, USA
3 tial applications are a main driver for its flourishing. These
University of Alcalá, Alcalá de Henares, Madrid, Spain
4
IMDEA-WATER Parque Tecnologico de Alcala, Madrid,
Spain
century to discover that bacteria can transfer electrons out/in
of the cell towards/from electrodes without the aid of these
artificial electron shuttling compounds [2–4]. This was the
real start of microbial electrochemistry, as discussed in more
detail by Schröder including a tutorial on how to get your
own electroactive biofilm anode [5]. Briefly, these electrodes
can act as electron donors (cathodes) [6] as well as acceptors
(anodes) [7] and the mechanism is denominated extracellu-
lar electron transfer (EET). Microorganisms with the physi-
ological capacity of EET are known as “electroactive” [8].
The archetypes of these electroactive microorganisms are
Shewanella oneidensis MR-1 and Geobacter sulfurreducens
that have been thoroughly studied [9–13]. Their pathways of
EET are depicted in Fig. 1—for more details please consult,
e.g., [8].
Thus, microbial electrochemistry has morphed into a
complex multidisciplinary area of its own. Apart from the
fascinating fundamentals, the possible advantages of poten-
applications are now summarized as microbial electro-
chemical technologies (MET) and span diverse fields, from

13
Vol.:(0123456789)
 4  Page 2 of 15 ChemTexts (2019) 5:4

wastewater treatment [15, 16], synthesis [17–19] and biosen-

sors [20] to restoration of polluted environments [21–25].
In the search to overcome technological bottlenecks for
the implementation of microbial electrochemical process in
real-field applications [26], new electrode designs and mate-
rials have been tested (Fig. 2) [27]. These include 3D-elec-
trodes, like felts and foams. Here especially bed electrodes
defined as electron conductors based on granules of conduc-
tive particles, have become of common use by numerous
researchers and are envisaged for several applications [28].

Bed electrode: an electrode made

of electrodes

Bed electrode reactors have been intensively studied, engi-

Fig. 1  Principal sketch of extracellular electron transfer mechanisms
on the example of anodic processes: a direct extracellular electron
transfer (DEET) based on physical contact of the cell surface redox
molecules and the electrode; b DEET via microbial nanowires; and
c mediated extracellular electron transfer (MEET) by redox-carriers
molecules that are reduced/oxidized at the cell surface or within the
cells. For details, see, e.g., [5, 14]
neered and used since the second half of the last century
in the frame of abiotic electrochemical processes [32–36].
The granular and porous nature of bed electrodes provides a
larger accessible reaction area and hence a higher active area
for biotic and abiotic reactions. This can also be exploited
for bioelectrochemical reactions and hence has boosted the
study of bed electrodes within the field of microbial electro-
chemistry in the last 10 years [37]. The high ratio between
active electrode area and reactor volume may bring the reac-
tor performance closer to the application or in other words

Fig. 2  Different carbon-based
materials used as electrodes in
MET: a graphite plate [29]. b
Carbon brush [29]. c Carbon
felt [29]. d Reticulated vitreous
carbon [30]. e Porous carbon
with defined pore size [31]. f
Granular activated carbon [29].
g Geobacter sulfurreducens
biofilms on a granular vitreous
carbon. h Structure of graphite
granules. i Porous structure
of activated carbon granules
(reproduction with permission
of respective publisher)

13
ChemTexts (2019) 5:4 Page 3 of 15  4
may allow MET surviving to the sword of Damocles of eco-
nomic viability.
The conductive granules used in bed electrodes can
remain either static [37, 38], or moving, i.e., using stir-
ring [39], recirculation [40] or fluidization of the granules
[41–44]. Figure 3 shows a fixed (3a) and a fluidized (3b) bed
electrode for laboratory studies. The assortment of granules
is electrochemically connected by an embedded electro-
chemically inert, but electrically conductive material called
current collector, usually made of metal or carbon (Table 1).
Fig. 3  Scheme of a a fixed and b a fluidized bed electrode reactor:
the bed electrode, made of granules, is acting as the working elec-
trode (WE). The three-electrode system is completed with a reference
electrode (RE) and the counter electrode (CE). Both bed electrode
reactors have an embedded current feeder/collector. Number code: (1)
recirculating flow; (2) flow distributor; (3) current feeder/collector
The approach of fluidized electrochemical bed electrode
reactors (Fig. 3b) is derived from traditional fluidized bed
reactors. These are extensively used in the field of water
treatment [54] for achieving high biomass production to treat
high loads of organic matter. In these configurations the bed
is composed of inert carriers, e.g., the mineral biolite, in
order to facilitate microbial adhesion. Nevertheless the fun-
dament of fluidization remains the same with all granular
material, conductive or inert, being fluidized by a controlled
and uniformly distributed upward flow of liquid. When mini-
mum fluidization velocity is obtained the electrode attains
fluid-like behavior [55]. The fluidization and hence mobil-
ity of the particles circumvents some constrains of fixed
biofilm-based electrodes, for instance including slow mass
transfer between solution and microorganisms and pH drop
at the electrode surface [56]. At the same time fluidization
provides good mass transfer and temperature distribution.
A first approach of fluidized bed electrodes was reported
using fluidized carbon granules acting as an anode for the
oxidation of an artificial soluble redox mediator [48]. Sub-
sequent, fluidized granules of activated carbon supported
the treatment of real brewery waste water [43, 44]. In 2017
also a fluidized anode made of glassy carbon granules sup-
ported the growth of G. sulfurreducens [42]. As thereby a
planktonic growth was achieved, it questioned the biofilm
paradigm of this electroactive model-organism.
Interestingly fixed-bed electrodes have been also inte-
grated in constructed wetlands [57], another established
technology for wastewater treatment, resulting in the so-
called ­METlands® (wetland plus a MET) [58]. ­METlands®
have shown to enhance the biodegradation rates during
wastewater treatment reducing the classical constructed
wetland dimensions and the phenomenon of clogging of the
filter substrate [15, 59]. The technology was demonstrated
of being capable to treat cubic meters of real wastewater for
over 2 years [60].
A plethora of carbon-based materials have been used for
bed electrodes (Table 1), including for instance granular
activated carbon [44, 53] and graphite granules [47, 61] as
well as biochar [51, 62] and coke [15]. Thereby especially
materials like biochar may become of importance for scal-
ing-up due to their significant economic and environmental
benefits [51].
Analysis of bed reactors: engineering focus
Different physical properties have to be considered for bed
electrode engineering. The main factors that determine the
performance of bed electrode reactors are hydrodynamic
properties and resistance. The latter is caused by the unequal
potential distribution in the bed electrode that is a result of
electric resistance between (and in) the particles.
13
 4 

Table 1  Selected literature on MET using bed electrodes: the bed electrode reactors are designated under different terms used in the original publications

13
Type of microbial electrochemical reactor Bed electrode material Current collector Applications/objectives References

Single-chamber tubular reactor Graphite granules (1.5 and 5 mm Ø) at the anode Graphite rod Waste water treatment with concomitant power [37]
Page 4 of 15

generation
Single-chamber reactor Graphite granules (2–3 mm Ø; 3.75 cm3 and Titanium wire Biosensor for measuring the organic content of [45]
12.5 cm3) at the anode wastewater
Single and double-chamber reactor Activated carbon granules (150 g) at the anode Graphite rod Waste water treatment with concomitant power [46]
generation
Single-chamber baffled air–cathode reactor Graphite granules (3–5 mm Ø) at the anode Graphite plate COD removal rate for high-concentration waste- [47]
water
Double-chamber reactor Graphite granules (2–7 mm Ø) at the anode and Carbon rod Reducing oxygen over-potential at the cathode [38]
cathode (500 cm3 in total)
Single-chamber anaerobic fluidized bed electrode Active carbon/graphite granules (0.2–0.9 mm Ø; Graphite rod Current production while simultaneously treating [48]
reactor 100 g) at the anode wastewater
Single-chamber cylindrical up-flow fixed-bed Activated carbon granules (1.5–1.7 mm Ø; Stainless steel mesh Degrading a reactive azo dye (Reactive Red 272) [49]
electrode reactor 142 cm3) at the cathode with a total surface area
of 664 m2 g−1
Double-chamber packed-bed air–cathode reactor Activated carbon granules, semi-coke, carbon felt Titanium mesh Material testing for increasing power generation [50]
cubes and graphite granules (5 g of granules
of each material, smaller than 1 cm Ø) at the
cathode
Double-chamber reactor Activated carbon, graphite and biochar granules at Titanium wire Power density and anodic material optimization [51]
the anode chamber (75 cm3 volume)
Double-chamber fluidized capacitive reactor Activated carbon granules at the anode (0.84– Graphite plates
Proof-of-principle of the fluidized capacitive [40]
1.18 mm Ø; 100 g) system
Double-chamber stirred reactor Activated carbon granules (0.5 g) Titanium mesh Testing the concept of intermittent discharge by [39]
suspended biofilm particles
Single-chamber cylindrical up-flow fixed-bed Activated carbon granules at the cathode (142 cm3) Stainless steel mesh Wastewater treatment processes for azo dye deg- [52]
electrode reactor radation
Single-chamber fluidized bed electrode reactor Glassy carbon granules (0.63–1 mm Ø; 80 cm3); Graphite plate Studying novel modes to culture electroactive [42]
graphite granules (0.42–0.69 mm Ø; 80 cm3) bacteria
Single-chamber microbial electrochemical wet land Coke granules (5–10 mm Ø; 0.006 m3) Graphite plate Integrating MET with natural wastewater treatment [15]
(METland) biofilters used in constructed wetlands
ChemTexts

Single-chamber fluidized bed electrode reactor Activated carbon granules (0.6–1 mm Ø ; 80 cm3) Graphite plate Integration of electrocoagulation with MET for [43]
treating a brewery wastewater
Single-chamber fluidized bed electrode reactor Activated carbon granules (0.6–1 mm Ø; 80 cm3) Graphite plate Waste water treatment [44]
Bioelectrochemical double-chamber reactors Activated carbon granules (specific surface area of Graphite plate Methane production [53]
764 m2 g−1; 1–3 mm Ø) and graphite granules
(2019) 5:4

(specific surface area of 0.438 m2 g−1; 3–5 mm

Ø) at the cathode
ChemTexts (2019) 5:4 Page 5 of 15  4
Both gradients, hydrodynamic and potential distribution
can be mathematically described. For example ohmic drop
(ΔEohmic) can be calculated as shown in Eq. (1) following
Ohm’s law to be equal to the product of the current (i) and
the ohmic or uncompensated resistance (Ru) [36, 63]:
ΔEohmic = i × Ru . (1)
Yet, one has to bear in mind that this formula, as the
hydrodynamics equations, e.g., Navier–Stokes formulas
[64], only hold true under the assumption of ideal scenario
(isotropic constant material properties (like resistance),
homogenous flow fields, etc.). This is not the case in bed
electrodes, especially in larger scale. Here a more empirical
approach is mostly needed.
From an engineering perspective the first of these vari-
ables, hydrodynamic properties, is more often considered as
key parameter for setting the operational conditions. Gener-
ally circulation of the process liquid that is also serving as
microbial medium improves the mass and heat transfer. It is
desirable to avoid process limitations, e.g., based on limited
ion or substrate diffusion, especially at the granules. The
kind and strength of this circulation is critical for the physi-
cal state (fluidized or fixed) of the bed electrode.
In general, selecting a specific reactor configuration and
mode of operation will depend on the kind of reaction one
aims to study. For instance, if the challenge is to purify a low
conductivity wastewater or low concentrations of a pollut-
ant, a design with an enhanced mass transfer (i.e., fluidized
bed electrode) may improve the performance of the pro-
cess [65]. Alternatively, supplying an external supporting
electrolyte (that means an electrochemically inert substance
that increases the ionic conductivity of the medium) such as
NaCl may help as well in this regard. However, both meas-
ures also have environmental disadvantages (power con-
sumption, addition of chemicals) that need to be balanced.
Design considerations for bed electrode reactors
Both the physical and the chemical characteristics of the
granular material and the bulk liquid will determine the con-
ditions for getting the bed electrode into a fluidized state.
There are numerous design parameters that can turn the con-
struction and starting-up stages into a complex endeavor.
For a proper bed electrode design one should take into con-
sideration at least the following characteristics: properties
of the granules (size, shape, density, mechanical resistance)
[66], properties of the bed electrode (particle size distribu-
tion, packing in the bed electrode), characteristics of the
fluid (viscosity and density) [67] and operational parameters
(for fluidized bed electrodes superficial velocity of the bulk
medium will control the height of the bed electrode) [42].
Fluidized bed electrodes typically use granules of smaller
size (see Table  1). Particle size selection for fixed-bed
electrodes relies rather on the optimization of the hydrau-
lic conditions. The smaller the particles, the higher is liq-
uid–electrode contact and hence the process kinetics, but at
the expense of decreased treatment capacity [68], and the
likability for clogging due to biomass accumulation or solids
deposition.
Laboratory-scale reactors based on granular electrodes
can be realized in very different architectures. For fixed-bed
electrode reactors, in order to optimize granule–medium
contact, there should be a well-designed fluid distribution
system, like tubular pipes, assuring laminar flow [69]. Rec-
tangular vessels with horizontal or vertical laminar flow,
e.g., used for M­ ETlands® [70], have also been successfully
employed for water purification purposes and are easier to
implement on practical scale. However, they are more likely
to hold dead zones characterized by poor medium transport
and thus reduced microbial activity [71]. Either one cham-
ber or two chamber configurations have been developed for
these fixed-bed electrode reactors [46, 70]. Fluidized bed
electrodes are characterized by a turbulent regime with
complete or partial mixing, depending on the recirculating
flow and mobility of the granules, allowing the utilization
of a wider range of reactor geometries. For a fluidized bed
electrode reactor the main component of the reactor design
is the discharging chamber. This is an electrochemical cell
where a current collector is placed for electrically interacting
with the recirculating particles [40]. However, fluidized bed
electrodes require specific design of more reactor compo-
nents, like the distribution system for the up-flowing elec-
trolyte that provokes the suspension of granules. Typically,
the recirculating flow enters from the bottom of the reactor
through a pipe network (annular, star-shaped) or a distrib-
uting plate [72] with nozzles directed towards the bottom
of the reactor which causes a high turbulence. As reactor
body, cylindrical columns have been typically used for these
configurations, similarly to widely used granulated wastewa-
ter treatment systems (like Expanded Granular Sludge Bed
(EGSB) or Upflow Anaerobic Sludge Blanket (UASB) [73]).
Operational and process parameters
One of the most desirable capabilities of any reactor is to be
able to instantly steer the performance by easily tuning one
or several parameters. This process control can be achieved
in MET by several means that are included in Table 2. Many
of them can only be applied to fluidized electrodes due to
the nature of the electrode design. For instance, an in situ
control of the system performance can be achieved by setting
the physical state of the fluidized bed electrode, i.e., varying
the frequency of particle contact, bed electrode expansion,
etc. Parameters related to medium composition or biological
aspects (inoculum, microbial growth control, etc.), for that
13
 4  Page 6 of 15 ChemTexts (2019) 5:4

Table 2  List of the main operational and process parameters for microbial electrochemical technologies using bed electrodes
System operational parameter Equipment or technique Short-term affected proper- Long-term affected properties Reference studies
needed ties

Electrode parameters/properties
 Anode or cathode potential/ Potentiostat/power source Biocatalysis kinetics (micro- Microbial communities distri- [74–76]
cell ­voltage#1,#2,#3 bial energy gain) bution
Current production
Product distribution
#1,#2,#3
 Electrode ­surface Electrode material addition or Biocatalysis kinetics Biomass quantity [77]
removal Current production
 Electrode surface-to-volume Recirculating pump Electrode surface-to-solution Biofilm thickness on particles [78, 79]
­ratio#2 Properties or quantity of volume ratio Biomass quantity
electrode material Biocatalysis kinetics
Current production
Electrochemical performance
 Inter-particle connection Recirculating pump Conductivity of bed elec- Biofilm thickness on particles [80, 81]
(frequency or number trodes Biomass quantity
particles collision)#2 Electrochemical performance
Biocatalysis kinetics
Current production
System parameters
 Hydraulic retention time Feeding pump Biocatalysis kinetics Biomass activity [15, 43]
(HRT)#1,#2,#3 Current production Microbial communities distri-
Biomass activity bution
 Organic ­load#1,#2,#3 Feeding pump Biocatalysis kinetics Microbial communities distri- [44, 82]
Substrate concentration in Current production bution
influent Product distribution Biomass activity
 Temperature#1,#2,#3 Heating system Biocatalysis kinetics Microbial communities distri- [83, 84]
Current production bution
Biomass activity
Product distribution
 Recirculation ­flow#1,#2,#3 Recirculating pump Biocatalysis kinetics Biomass activity [42, 85]
Current production Microbial communities distri-
bution
 Electrode volume or sur- Recirculating pump Biocatalysis kinetics Biofilm thickness on particles [40, 86]
face/reactor volume ­ratio#2 Current production

The superscripts within the column 1 indicate the type of electrodes that can be modify/tuned for the different system operational parameters
(#1: fixed-bed electrodes #2: fluidized-bed electrodes #3: non-granular electrodes)

the control is out the scope of a pure engineering focus, have
not been considered in Table 2.
Abiotic electrochemical characterization
of bed electrodes
A first approach for assessing the microbial electrochemi-
cal properties of a bed electrode reactor can be performed
using a redox probe (i.e., a chemical compound that can
be reversibly oxidized and reduced within the range of the
potentials used for the biotic experiment). Here the redox
reaction of Fe(II)/Fe(III) in ferri-/ferrocyanide is the most
often used for testing the abiotic electrochemical proper-
ties. By employing techniques such as chronoamperometry
or cyclic voltammetry, one can obtain information on, for
instance, system limitations (electrochemical resistances),
influence of operational parameters and obtain reference
values for future benchmarking of operation as microbial
electrodes. This kind of assay can be applied to both fixed
and fluidized bed electrode reactors. Figure  4 shows an
example of the dependency of the current production and
expansion of the bed electrode on linear velocity of the recir-
culating liquid (ratio between recirculating flow rate and
section of the reactor) [42]. It can be seen that a current,
here ~ 250 A m−3 is obtained under no fluidization state of
the bed electrode. As soon as the recirculation in the system
starts to flow, the current density is substantially enhanced
up to almost 1500 A m−3 for full fluidization. This illustrates
the positive effect of the mixing on the electrochemical reac-
tion. As the bed electrode is expanded, the current density
produced from Fe(II) oxidation to Fe(III) linearly increases
with superficial velocity of the recirculating flow (but not
linear with expansion of the bed electrode) until a certain

13
ChemTexts (2019) 5:4 Page 7 of 15  4
Fig. 4  Current production and expansion of bed electrode as function
of the linear velocity of the fluid of a bed electrode reactor: The redox
probe used in the study was an equimolar solution of 1  mM ferro-/
ferricyanide in an aqueous 1  M KCl solution (figure extracted from
[42]—reproduction with permission of respective publisher). Current
collector at the bottom is not depicted in the superior figure
value. Beyond that the current starts to decrease (not shown
in Fig. 4). This effect can be due to a loss of electrical con-
ductivity along the bed electrode. At this point, the expan-
sion of the bed electrode is so high that the inter-particle
distances within the bed electrode are too large to support
rapid electron transfer. Consequently, this high internal elec-
trochemical resistance of the fluidized electrode is negatively
affecting the electrochemical reaction rate. Since the energy
consumption of a reactor will directly depend on the power
demand of the recirculating pump, this kind of abiotic assay
can be useful for selecting adequate operating conditions for
a desired reaction.
However, in general one should expect lower current den-
sity when operating the system under biological conditions
than for abiotic operation. First, microbial reactions occur
usually at lower rates and, second, the active (or “accessi-
ble”) electrode area available for microorganisms is typically
lower than for abiotic electrochemical reactions. This can be
explained by the inaccessibility of nano- and micro-pores
to microbial cells [87]. Nonetheless, a first abiotic electro-
chemical characterization of a bed electrode reactor can be
a potential tool for optimizing the design of a given system
and for an early identification of any possible failure, includ-
ing, for instance, weak electrical connections, low electrode
surface area and short-circuits. Further, the electrochemistry
of single granules can be assessed even when operated as
MET (see “Bed electrodes: 3D redox mosaics”).
Another aspect to take into account with bed electrodes is
their high internal resistance, not negligible compared to that
of some other electrodes, e.g., planar electrodes. This high
internal resistance is based on the poor and only pointwise
electrical contact between the granules leading to the forma-
tion of potential gradients. This results in a poor polarization
behavior meaning that the electrochemical potential cannot
be set identical for all particles and hence the entire bed
electrode. This high resistance does already play a role in
the case for setting a constant potential, like in chronoamper-
ometry, during electrochemical cultivation, but is even more
a constraint when changing the electrode potential in time,
as, e.g., in cyclic voltammetry; see Fig. 6d. With no doubt
the poor polarization behavior resulting in a heterogeneous
potential distribution will have an influence in the microbial
selection as explained in the next chapter.
Bed electrodes as microbial collage
As discussed just above, due to high internal resistance
during electrochemical cultivation a potential gradient is
established through the bed electrode procuring a plethora
of redox scenarios within the same bed electrode reactor
(Fig. 5) [88, 89]. This potential gradient may trigger differ-
ent EET mechanisms of the same microorganisms [90]. For
instance, electroactive model bacteria as Shewanella onei-
densis exhibit a different exoelectrogenic physiology when
the electrode potential changes [91]. Similarly, depending
on the anode potential G. sulfurreducens shows different
electron transport pathways that are used [92, 93]. More
importantly, this redox mosaic provides different ecologi-
cal niches that hence may foster electrochemically driven
microbial selection. Thus, bed electrodes are optimal scene
plots to study the influence of the electrode potential on
microbial community dynamics and the establishment of
trophic chains.
Analyzing the entire collection of microorganisms in a
specific microbiome can reveal structure–function relation-
ships allowing proactive microbiome management [94]. At
present very little is known about the microbiome at elec-
trodes, although a diverse arsenal of molecular techniques
for phylogenetic and functional level characterization
exists [14]. Likewise only few studies have been devoted
to identify the microorganism in bed electrodes and even
lesser to deepen the knowledge on the relationship between
redox electrode potential and ecological function of the
13
 4  Page 8 of 15 ChemTexts (2019) 5:4

Fig. 5  Gradients in fixed-bed electrodes: the high internal resistance the end of the bed electrode. This redox gradient combined with com-
between the granules does not allow the establishment of a homo- plex inoculum and complex medium, e.g., wastewater, drives micro-
geneous redox potential in the bed electrode. This poor polarization bial selection and formation of trophic chains
behavior provokes a redox gradient from the current collector through

Fig. 6  Analysing bed electrodes

using cyclic voltammetry: a
electroactive microorganisms
performing EET to granules
that are b part of a fixed-bed
electrode reactor MET. c
Photograph of a microbial
electrochemical reactor with an
e-Clamp with outset: zoom on
the e-Clamp electrode. d CV
of a Geobacter based fixed-
bed electrode. The undefined
shape, due to the high internal
resistance, impedes inferring
any information on the underly-
ing EET mechanisms; e this
information can be gained with
CV of a single graphite granule
with a Geobacter-based biofilm
(reproduction of c) with permis-
sion of respective publisher

microorganisms associated to conductive granules. For
instance, 16S rRNA gene sequencing has been used with the
aim of understanding the role of anodic [15] and cathodic
microbial populations [53]. This 16S rRNA gene sequenc-
ing procedures provides information about the phylogenetic
composition termed operational taxonomic units (OTU)—or
plainly but not entirely correct the “species”—of a microbial
community. It can be applied for assessing the evolution of
microbiomes over time and under changing environmental or
operational conditions (i.e., different electrode potential or

13
ChemTexts (2019) 5:4 Page 9 of 15  4
organic loads in the reactor). Another molecular technique is
fluorescence in situ hybridization (FISH) being single cell-
based approaches, allowing spatially resolved microscopic
imaging. This method has revealed a microbial stratifica-
tion in the biofilms developed on activated carbon granules
of a fluidized bed electrode reactor [44]. Geobacter spe-
cies—that are performing DEET—were highly enriched in
the innermost layers of the biofilm (thickness of 10 mm),
which were in contact with the conductive particles. The
other share of bacteria was located in the outermost layers.
This strongly underlines the competitive advantage of Geo-
bacter over other microbial species for respiring at an anode.
As a consequence Geobacter and other highly electroactive
microorganisms may possess a key role by connecting non-
electroactive microorganisms involved in different degrada-
tion processes (i.e., microorganisms that generate electron
donors like acetic acid from breaking down complex organic
matter) and the electrode.
Bed electrodes: 3D redox mosaics
Dynamic electrochemical techniques, e.g., cyclic voltam-
metry [95], are of common use to obtain thermodynamic
information about the EET. In CV the current is measured
as function of a changing electrode potential. This is usually
done using a three-electrode setup and at a certain rate of
potential change (scan rate, mV s−1). However, CV does not
provide any thermodynamically valuable information when
performed on entire bed electrodes (Fig. 6d). The already
explained poor polarization behavior of and hence potential
distribution in the bed electrode impedes the establishment
of a fixed and known potential throughout the whole bed
electrode, especially when this potential changes over time.
Hence very little is known on microbial EET in bed elec-
trodes. To tackle this challenge an alternative method has
been presented, based on the CV analysis of single gran-
ules derived from a bed electrode [96]. This is achieved by
using a clamp (denominated e-Clamp from now on) that
acts as a holder contacting the granule with a conductive
wire serving as (electrochemically inert) current collector
(Fig. 6c). In other words, the granule that was part of a bed
electrode (Fig. 6b) is analyzed as a single electrode. Follow-
ing this strategy Rodrigo Quejigo et al. in [22] gained valu-
able information on EET of single granules derived from
Geobacter-based bed electrode (Fig. 6e) [96]. A previous
study described the current production of a single granule
reactor [97]. Yet, the biofilm development on these gran-
ule did not take place as part of bed electrodes, but as an
independent one single-granule electrochemical cell. Liter-
ally, the e-Clamp offers the possibility to “grab” temporarily
the granules from a bed electrode of any size and allows
its immediate electrochemical characterization. Therefore,
this tool that can be self-made using, e.g., a 3D-printing
approach, opens the possibility to survey the electrochemical
behavior of microbial communities on granulated electrodes.
In addition, this strategy to analyze single conductive parti-
cles can be adapted further, e.g., for studying the influence of
redox potentials on microbial trophic relations or the inter-
species electron transfer processes [98].
Case studies
To guide the reader and potential operator two hypothetical
case studies—either fluidized (case study A) or fixed (case
study B)—are discussed, illustrating general operational pro-
cedures of bed electrodes for MET.
Case study A: analysis of biofilm growth
and electroactivity in a fluidized bed electrode
reactor at different hydraulic scenarios treating
winery effluents
Biofilm formation is a complex process that is affected by
many factors such as characteristics and density of micro-
organisms, the material surface and the environmental con-
ditions, e.g., the hydrodynamics [99]. The rate of biomass
attachment on fluidized particles depends largely on the
velocity characteristics of the recirculating liquid in a fluid-
ized bed electrode reactor. Theoretically, a fluid motion in
the bulk liquid favors bacterial adhesion due to the enhance-
ment of the cell transportation to the surface by convection
[100]. However, shear forces can also reduce adhesion and
provoke cell detachment [101]. Additionally, the expansion
of the bed electrode that is affected by an increasing recir-
culating velocity of the liquid as well the conductivity along
the bed electrode determines the performance. Increasing the
inter-particle distance diminishes the charge transfer across
the bed electrode, whereas increasing the contact frequency
among the particles positively affects the electrical conduc-
tivity of the bed electrode. Thus, the hydraulic conditions in
the fluidized reactors strongly influence the particles colo-
nization (quantitative and qualitatively), and ultimately the
microbial electroactivity.
Fluidized bed electrode reactors made of activated car-
bon particles have been shown to effectively treat agro-
industrial wastewater [43]. For the present case study, a
reactor consisting of a bed electrode of activated porous
carbon particles is inoculated with a mixed culture adapted
to degrade anaerobically the organic matter contained in
the effluent from a winery. Biomass adaptation and biofilm
growth on the particles of the fluidized anode polarized
at + 0.2 V (vs. Ag/AgCl) leads to stable reactor perfor-
mances [removal of over 70–80% of Total Organic Car-
bon (TOC)]. Thereafter, an assessment of the effect of
13
 4  Page 10 of 15
varying expansion of the bed electrode on the developed
biofilm is performed. The different expansions of the bed
electrode are achieved by tuning the flow of the recircu-
lating pump, and therefore the superficial velocity (ratio
between recirculating flow rate and section of the reac-
tor) in the reactor. Four different expansions of the bed
electrode are proposed to be tested, and samples (in trip-
licates) of the anodic particles are gained for studying the
biofilm growth at each condition. The strategy to address
this research consists of analysing each sample for three
different aspects: biofilm characterization, electrochemical
performance and organic matter degradation rate. First,
physical biofilm properties are examined by microscopic
imaging and thermogravimetric analysis (measurements
of biofilm mass over time as a function of temperature).
This approach provides quantitative data such as biofilm
thickness, coverage and average biofilm mass per granule
or biofilm density. Scanning electron microscopy (SEM)
images allow obtaining morphological information of
the biofilm surface [102]. By imaging with a fluorescent
confocal microscopy one can obtain the biofilm average
thickness and metabolic structure (like cell viability) [103,
104]. The probe DAPI (4′-6-diamidino-2-phenylindole)
can be employed for staining all the biomass and provide
quantitative information (biofilm thickness and number
of total bacteria). Additionally, a LIVE/DEAD BacLight
Bacterial Viability Kit can be utilized for analyzing the
metabolic spatial activity within the biofilm [105]. Ther-
mogravimetric analysis allows to quantify the mass of
biofilm developed on a certain volume of bed electrode
[41, 106]. Secondly, the electrochemical performance can
be determined by measuring stationary current density
(chronoamperometry at (different) fixed potentials), cell
potential and the polarization curves of the fluidized bed
electrode. Finally, the degradation capacity of the reactor
at each condition can be examined by performing chemi-
cal analyses of the effluent samples such as total organic
carbon (TOC), chemical oxygen demand (COD), volatile
fatty acids (VFA), or nutrients levels (nitrogen and phos-
phorous), following standard methods [107]. By correlat-
ing this data with the total charge harvested, the coulombic
efficiency (fraction of electrons from the oxidation utilized
by bacteria for current production) can be estimated for
each scenario (Eq. 2):
mol of electrons harvested as current ∫tt idt
CE = = (
0
) ,
mol of electrons in removed COD nFV CODin − CODout ∕MO2
where n is the number of electrons exchanged per
mole of oxygen (4 electrons), F is Faraday constant
(96.485 C mol−1), V is the net volume of the reactor (L),
­CODin and C
­ ODout are the COD concentrations in influent
13
ChemTexts (2019) 5:4
and effluent, respectively, and MO2 is the molecular weight
of ­O2 (32 g mol−1) [16].
As a result of this study, valuable information for correlat-
ing bed electrode expansion with biofilm quantity/thickness,
biofilm viability, microbial electrochemical performance and
the reactor water purification capacity can be obtained. Fur-
ther, it is possible to obtain qualitative data regarding the
shear stress environment at which the biofilm attached to
the fluidized particles is exposed when operating at a spe-
cific expansion of the bed electrode. The loss of biomass
attached, and therefore, microbial activity (loss on COD
removal capacity of the fluidized bed electrode reactor),
can be a sign of a non-favorable reactor operation, as too
strong mixing leads to strong (“aggressive”) inter-particle
collisions. Overall, this case study shows how to optimize
the performance of the fluidized bed electrode reactor. Addi-
tionally, one can obtain helpful data for balancing the puri-
fication capacity with the energy demand of the treatment.
Since the energy consumption of the global process will
largely depend on the energy requirements of recirculating
pumping it is critical to operate at a minimum expansion of
the bed electrode, but without compromising the kinetics of
the on-going microbial electrochemical reactions, which is
controlled by the quantity and quality of the biofilm in the
reactor.
Case study B: removal of residual chlorinated
pesticide from the effluent of a wastewater
treatment plant in a fixed‑bed electrode reactor
Chlorinated aliphatic hydrocarbons, like trichloroethene
(TCE), are contaminants that can be found in industrial
effluents. Among other methods for eliminating this pol-
lutant, microbial reductive dehalogenation is known as
one the most effective bioremediation processes [108]. For
this biological reaction an electron donor is needed such
as hydrogen, organic matter or a cathode as those halogen-
ated pollutants commonly act as terminal electron acceptors.
For our case study the target effluent comes from a biologi-
cal treatment in a wastewater treatment plant consisting of
two sequential anaerobic/aerobic reactors, which could not
remove TCE below 25 µmol L−1. As treatment to degrade
TCE, a fixed-bed electrode reactor is proposed as MET have
been shown to effectively reduce halogenated compounds in
(2)
the cathode under anaerobic conditions [109, 110]. One of
the parameters to optimize the performance is the quantity
of bed electrode needed to meet the specific legal require-
ments of effluent quality. Additionally, as illustrated more
ChemTexts (2019) 5:4 Page 11 of 15  4
Fig. 7  Proposed system for studying substrate, potential and removal
rates as well as gradients along a fixed-bed electrode under non-mix-
ing conditions, as discussed for case study B
generally above, a low conductivity of the electrode and low
ionic conductivity of the electrolyte causes a potential drop
along the bed electrode and creates bioelectrochemically
inactive areas that poorly contribute to the cathodic reduc-
tion of TCE. Thus, microbial electrochemical post-treatment
is recommended. To analyze the kinetics of TCE-reduction
along a cathodic fixed-bed electrode in order to estimate
both, bed electrode and reactor size, configuration consist-
ing of a column continuously fed through a flux distributor
at the bottom and effluent outlet located at the top of the
column so that laminar flow conditions across the column
are established (Fig. 7). Certainly, like every MET this kind
of reactor has to be a compromise between biological and
electrochemical needs and application constraints and hence
does not fulfill the conditions of each fully.
The first two-thirds of the column are filled with granular
graphite (working electrode, acting as cathode), electrically
separated (i.e., with non-electrically conductive particles)
from a granular fixed-bed anode (counter electrode) located
at the other top third of the reactor. Each electrode is sup-
ported on a circular stainless steel mesh acting as current
collector. A cathode potential of − 0.7 V (vs. Ag/AgCl) is
set throughout the entire experiment. Different hydraulic
retentions times (calculated from net reactor volume), being
increased gradually, are tested with a fixed concentration of
TCE (i.e., 25 µmol L−1). The column should include numer-
ous vertical and aligned ports distributed along the fixed-bed
cathode in order to sample the medium at different heights
of the bed electrode. Furthermore, in the same way electrical
connections (like wire ports) and reference electrodes ports
should be included in the design in order to be able to meas-
ure the cathode potential at each sampling port. An example
of a possible design is shown in Fig. 7 allowing the study of
concentration, potential and reaction rate profiles. Further
electrochemical assays can provide information about elec-
trochemical limitations to current consumption like deple-
tion of the reactant (TCE), build-up of the reaction products
(TCE-reduction sub-products) [111] or polarization char-
acteristics. Additionally, a deeper analysis at single-granule
level can be gained at different bed heights [96] both from an
electrochemical (see “Bed electrodes: 3D redox mosaics”)
and substrate degradation capacity standpoint.
Conclusions
Overall, bed electrodes are complex 3D-electrochemical
environments that provide unprecedented possibilities for
achieving high space–time yields and volumetric rates in
MET. Bed electrodes possess numerous limitations that
can be avoided, if proper experimentation (or simulation if
parameters are available) is executed. The main take-home
messages for deciding on bed electrodes versus planar elec-
trodes are summarized in Fig. 8. Certainly further options of
3D-electrodes like brushes [112] or sponges [113] do exist
that are beyond the scope of this article.
13
 4  Page 12 of 15
Fig. 8  Flowchart for selection of the most adequate electrode type (planar versus bed electrodes) and its operation modus (fluidized versus fixed-
bed electrodes) for MET: the yes or no answers to the questions will guide to the most suitable type of reactor
Acknowledgements FH acknowledges support by the BMBF
(Research Award “Next generation biotechnological Processes—Bio-
technology 2020+”) and the Helmholtz-Association (Young Investiga-
tors Group). This work was supported by the Helmholtz-Association
within the Research Programme Renewable Energies.
References
1. Potter MC (1911) Electrical effects accompanying the decom-
position of organic compounds. Proc R Soc Lond Ser B
84(571):260–276
2. Lovley DR, Phillips EJP (1988) Novel mode of microbial energy
metabolism: organic carbon oxidation coupled to dissimila-
tory reduction of iron or manganese. Appl Environ Microbiol
54(6):1472–1480
3. Myers CR, Nealson KH (1988) Bacterial manganese reduction
and growth with manganese oxide as the sole electron acceptor.
Science 240(4857):1319–1321. https​://doi.org/10.1126/scien​
ce.240.4857.1319
4. Schröder U (2011) Discover the possibilities: microbial bio-
electrochemical systems and the revival of a 100-year-old
13
ChemTexts (2019) 5:4
discovery. J Solid State Electrochem 15(7):1481–1486. https​://
doi.org/10.1007/s1000​8-011-1395-7
5. Schröder U (2018) A basic introduction into microbial fuel cells
and microbial electrocatalysis. ChemTexts 4(4):19. https​://doi.
org/10.1007/s4082​8-018-0072-1
6. Rosenbaum M, Aulenta F, Villano M, Angenent LT (2011)
Cathodes as electron donors for microbial metabolism: which
extracellular electron transfer mechanisms are involved? Biore-
sour Technol 102(1):324–333. https​://doi.org/10.1016/j.biort​
ech.2010.07.008
7. Kim BH, Kim HJ, Hyun MS, Park DH (1999) Direct electrode
reaction of Fe(III)-reducing bacterium, Shewanella putrefaciens.
J Microbiol Biotechnol 9:127–131
8. Koch C, Harnisch F (2016) Is there a specific ecological niche
for electroactive microorganisms? ChemElectroChem 3(9):1282–
1295. https​://doi.org/10.1002/celc.20160​0079
9. Carmona-Martínez AA, Harnisch F, Kuhlicke U, Neu TR,
Schröder U (2013) Electron transfer and biofilm formation of
Shewanella putrefaciens as function of anode potential. Bioel-
ectrochemistry 93:23–29. https​://doi.org/10.1016/j.bioel​echem​
.2012.05.002
10. Lovley DR (2011) Live wires: direct extracellular electron
exchange for bioenergy and the bioremediation of energy-related
ChemTexts (2019) 5:4 Page 13 of 15  4
contamination. Energy Environ Sci 4(12):4896–4906. https:​ //doi.
org/10.1039/C1EE0​2229F​
11. Estevez-Canales M, Kuzume A, Borjas Z, Fueg M, Lovley D,
Wandlowski T, Esteve-Nunez A (2015) A severe reduction in the
cytochrome C content of Geobacter sulfurreducens eliminates
its capacity for extracellular electron transfer. Environ Microbiol
Rep 7(2):219–226. https​://doi.org/10.1111/1758-2229.12230​
12. Busalmen JP, Esteve-Núñez A, Berná A, Feliu JM (2008) C-type
cytochromes wire electricity-producing bacteria to electrodes.
Angew Chem Int Ed 47(26):4874–4877
13. Esteve-Núnez A, Busalmen JP, Berná A, Gutiérrez-Garrán C,
Feliu JM (2011) Opportunities behind the unusual ability of Geo-
bacter sulfurreducens for exocellular respiration and electricity
production. Energy Environ Sci 4(6):2066–2069
14. Koch C, Korth B, Harnisch F (2017) Microbial ecology-
based engineering of microbial electrochemical tech-
nologies. Microb Biotechnol 11(1):22–38. https ​ : //doi.
org/10.1111/1751-7915.12802​
15. Aguirre-Sierra A, Bacchetti-De Gregoris T, Berná A, Salas JJ,
Aragón C, Esteve-Núñez A (2016) Microbial electrochemical
systems outperform fixed-bed biofilters in cleaning up urban
wastewater. Environ Sci Water Res Technol 2(6):984–993. https​
://doi.org/10.1039/c6ew0​0172f​
16. Logan BE, Rabaey K (2012) Conversion of wastes into bioelec-
tricity and chemicals by using microbial electrochemical tech-
nologies. Science 337(6095):686–690
17. Hegner R, Rosa LFM, Harnisch F (2018) Electrochemical C ­ O2
reduction to formate at indium electrodes with high efficiency
and selectivity in pH neutral electrolytes. Appl Catal B 238:546–
556. https​://doi.org/10.1016/j.apcat​b.2018.07.030
18. Jourdin L, Freguia S, Flexer V, Keller J (2016) Bringing high-
rate, ­CO2-based microbial electrosynthesis closer to practical
implementation through improved electrode design and operat-
ing conditions. Environ Sci Technol 50(4):1982–1989
19. Patil SA, Arends JB, Vanwonterghem I, Van Meerbergen J, Guo
K, Tyson GW, Rabaey K (2015) Selective enrichment establishes
a stable performing community for microbial electrosynthesis of
acetate from ­CO2. Environ Sci Technol 49(14):8833–8843
20. Kretzschmar J, Böhme P, Liebetrau J, Mertig M, Harnisch F
(2018) Microbial electrochemical sensors for anaerobic digestion
process control–performance of electroactive biofilms under real
conditions. Chem Eng Technol 41(4):687–695
21. Rodrigo Quejigo J, Dörfler U, Schroll R, Esteve-Núñez A (2016)
Stimulating soil microorganisms for mineralizing the herbicide
isoproturon by means of microbial electroremediating cells.
Microb Biotechnol 9(3):369–380
22. Pous N, Balaguer MD, Colprim J, Puig S (2017) Opportunities
for groundwater microbial electro-remediation. Microb Biotech-
nol 11(1):119–135. https​://doi.org/10.1111/1751-7915.12866​
23. Rodrigo Quejigo J, Domínguez-Garay A, Dörfler U, Schroll R,
Esteve-Núñez A (2018) Anodic shifting of the microbial com-
munity profile to enhance oxidative metabolism in soil. Soil
Biol Biochem 116:131–138. https​: //doi.org/10.1016/j.soilb​
io.2017.09.012
24. Dominguez-Garay A, Rodrigo Quejigo J, Dorfler U, Schroll
R, Esteve-Nunez A (2017) Bioelectroventing: an electrochem-
ical-assisted bioremediation strategy for cleaning-up atrazine-
polluted soils. Microb Biotechnol 11(1):50–62. https​://doi.
org/10.1111/1751-7915.12687​
25. Rodrigo Quejigo J, Boltes K, Esteve-Nuñez A (2014) Microbial-
electrochemical bioremediation and detoxification of dibenzo-
thiophene-polluted soil. Chemosphere 101:61–65
26. Aulenta F, Puig S, Harnisch F (2018) Microbial electrochemi-
cal technologies: maturing but not mature. Microb Biotechnol
11(1):18–19
27. Kerzenmacher S (2017) Engineering of microbial electrodes. Adv
Biochem Eng Biotechnol. https​://doi.org/10.1007/10_2017_16
28. Yu YY, Zhai DD, Si RW, Sun JZ, Liu X, Yong YC (2017) Three-
dimensional electrodes for high-performance bioelectrochemical
systems. Int J Mol Sci 18(1):90. https​://doi.org/10.3390/ijms1​
80100​90
29. Wei J, Liang P, Huang X (2011) Recent progress in electrodes
for microbial fuel cells. Bioresour Technol 102(20):9335–9344.
https​://doi.org/10.1016/j.biort​ech.2011.07.019
30. Wei W, Zhang Y, Komorek R, Plymale A, Yu R, Wang B, Zhu Z,
Liu F, Yu XY (2017) Characterization of syntrophic Geobacter
communities using ToF-SIMS. Biointerphases 12(5):05g601.
https​://doi.org/10.1116/1.49868​32
31. Chen X, Cui D, Wang X, Wang X, Li W (2015) Porous carbon
with defined pore size as anode of microbial fuel cell. Biosens
Bioelectron 69:135–141
32. Hiddleston J, Douglas A (1970) Current/potential relationships
and potential distribution in fluidized bed electrodes. Electrochim
Acta 15(3):431–443
33. Hiddleston JN, Douglas AF (1968) Fluidized bed electrodes—
fundamental measurements and implications. Nature 218:601.
https​://doi.org/10.1038/21860​1a0
34. Berent T, Mason R, Fells I (1971) Fluidised-bed fuel-cell elec-
trodes. J Appl Chem Biotechnol 21(3):71–76
35. Sedahmed G (1996) Mass transfer behaviour of a fixed bed elec-
trochemical reactor with a gas evolving upstream counter elec-
trode. Can J Chem Eng 74(4):487–492
36. Ibl N (1983) Current distribution. Comprehensive treatise of
electrochemistry. Springer, New York, pp 239–315
37. Rabaey K, Clauwaert P, Aelterman P, Verstraete W (2005)
Tubular microbial fuel cells for efficient electricity generation.
Environ Sci Technol 39(20):8077–8082. https:​ //doi.org/10.1021/
es050​986i
38. Tran HT, Ryu JH, Jia YH, Oh SJ, Choi JY, Park DH, Ahn DH
(2010) Continuous bioelectricity production and sustainable
wastewater treatment in a microbial fuel cell constructed with
non-catalyzed granular graphite electrodes and permeable
membrane. Water Sci Technol 61(7):1819–1827. https​://doi.
org/10.2166/wst.2010.140
39. Liu J, Zhang F, He W, Zhang X, Feng Y, Logan BE (2014)
Intermittent contact of fluidized anode particles containing
exoelectrogenic biofilms for continuous power generation in
microbial fuel cells. J Power Sources 261:278–284. https​://doi.
org/10.1016/j.jpows​our.2014.03.071
40. Deeke A, Sleutels TH, Donkers TF, Hamelers HV, Buisman CJ,
Ter Heijne A (2015) Fluidized capacitive bioanode as a novel
reactor concept for the microbial fuel cell. Environ Sci Technol
49(3):1929–1935. https​://doi.org/10.1021/es503​063n
41. Tejedor-Sanz S, de Gregoris TB, Salas JJ, Pastor L, Esteve-
Núñez A (2016) Integrating a microbial electrochemical system
into a classical wastewater treatment configuration for removing
nitrogen from low COD effluents. Environ Sci Water Res Technol
2(5):884–893
42. Tejedor-Sanz S, Quejigo JR, Berna A, Esteve-Nunez A (2017)
The planktonic relationship between fluid-like electrodes and
bacteria: wiring in motion. ChemSusChem 10(4):693–700. https​
://doi.org/10.1002/cssc.20160​1329
43. Tejedor-Sanz S, Ortiz JM, Esteve-Núñez A (2017) Merging
microbial electrochemical systems with electrocoagulation pre-
treatment for achieving a complete treatment of brewery waste-
water. Chem Eng J 330:1068–1074. https​://doi.org/10.1016/j.
cej.2017.08.049
44. Tejedor-Sanz S, Fernández-Labrador P, Hart S, Torres CI,
Esteve-Núñez A (2018) Geobacter dominates the inner lay-
ers of a stratified biofilm on a fluidized anode during brewery
13
 4  Page 14 of 15
wastewater treatment. Front Microbiol. https​://doi.org/10.3389/
fmicb​.2018.00378​
45. Di Lorenzo M, Curtis TP, Head IM, Velasquez-Orta SB,
Velasquez-Orta SB, Scott K (2009) A single chamber packed
bed microbial fuel cell biosensor for measuring organic content
of wastewater. Water Sci Technol 60(11):2879–2887. https:​ //doi.
org/10.2166/wst.2009.699
46. Jiang D, Li B (2009) Granular activated carbon single-chamber
microbial fuel cells (GAC-SCMFCs): a design suitable for large-
scale wastewater treatment processes. Biochem Eng J 47(1):31–
37. https​://doi.org/10.1016/j.bej.2009.06.013
47. Feng Y, Lee H, Wang X, Liu Y, He W (2010) Continuous elec-
tricity generation by a graphite granule baffled air–cathode
microbial fuel cell. Bioresour Technol 101(2):632–638. https​://
doi.org/10.1016/j.biort​ech.2009.08.046
48. Kong W, Guo Q, Wang X, Yue X (2011) Electricity genera-
tion from wastewater using an anaerobic fluidized bed microbial
fuel cell. Ind Eng Chem Res 50(21):12225–12232. https​://doi.
org/10.1021/ie200​7505
49. Cardenas-Robles A, Martinez E, Rendon-Alcantar I, Frontana
C, Gonzalez-Gutierrez L (2013) Development of an activated
carbon-packed microbial bioelectrochemical system for azo
dye degradation. Bioresour Technol 127:37–43. https​://doi.
org/10.1016/j.biort​ech.2012.09.066
50. Zhang X, Shi J, Liang P, Wei J, Huang X, Zhang C, Logan BE
(2013) Power generation by packed-bed air–cathode micro-
bial fuel cells. Bioresour Technol 142:109–114. https​://doi.
org/10.1016/j.biort​ech.2013.05.014
51. Huggins T, Wang H, Kearns J, Jenkins P, Ren ZJ (2014) Bio-
char as a sustainable electrode material for electricity produc-
tion in microbial fuel cells. Bioresour Technol 157:114–119.
https​://doi.org/10.1016/j.biort​ech.2014.01.058
52. Gonzalez-Gutierrez L, Frontana C, Martinez E (2015) Upflow
fixed bed bioelectrochemical reactor for wastewater treatment
applications. Bioresour Technol 176:292–295. https​: //doi.
org/10.1016/j.biort​ech.2014.11.018
53. Liu D, Roca-Puigros M, Geppert F, Caizan-Juanarena L, Na
Ayudthaya SP, Buisman C, Ter Heijne A (2018) Granular car-
bon-based electrodes as cathodes in methane-producing bio-
electrochemical systems. Front Bioeng Biotechnol 6:78. https​
://doi.org/10.3389/fbioe​.2018.00078​
54. Nicolella C, Van Loosdrecht M, Heijnen J (2000) Wastewa-
ter treatment with particulate biofilm reactors. J Biotechnol
80(1):1–33
55. Gupta CK, Sathiyamoorthy D (1998) Fluid bed technology in
materials processing. CRC Press, Boca Raton
56. Scott K, Yu EH (2015) Microbial electrochemical and fuel
cells: fundamentals and applications. Woodhead Publishing,
Newcastle University, UK
57. Wu H, Zhang J, Ngo HH, Guo W, Hu Z, Liang S, Fan J, Liu H
(2015) A review on the sustainability of constructed wetlands
for wastewater treatment: design and operation. Bioresour
Technol 175:594–601
58. Ramírez-Vargas CA, Prado A, Arias CA, Carvalho PN, Esteve-
Núñez A, Brix H (2018) Microbial electrochemical technolo-
gies for wastewater treatment: principles and evolution from
microbial fuel cells to bioelectrochemical-based constructed
wetlands. Water 10(9):1128
59. Aguirre A (2017) Integrating microbial electrochemical sys-
tems in constructed wetlands, a new paradigm for treating
wastewater in small communities. Universidad de Alcalá
60. Esteve-Núñez  A, Fernandez Ontivero J, Salas JJ, Berna
A, Reija Maqueda A, Aragon C, Aguirre-Sierra MA, Bac-
chetti de Gregoris T, Esteve-Núñez R, Barroeta B, Pidre JR
(2014) Long-term demonstration of a bioelectrochemically
constructed wetland for urban wastewater treatment. In:
13
ChemTexts (2019) 5:4
Conference: 11th IWA leading edge conference on water and
wastewater technologies LET 2014’, Abu Dabi
61. Rabaey I, Ossieur W, Verhaege M, Verstraete W (2005) Con-
tinuous microbial fuel cells convert carbohydrates to electric-
ity. Water Sci Technol 52(1–2):515–523
62. Huggins TM, Pietron JJ, Wang H, Ren ZJ, Biffinger JC (2015)
Graphitic biochar as a cathode electrocatalyst support for
microbial fuel cells. Bioresour Technol 195:147–153. https​://
doi.org/10.1016/j.biort​ech.2015.06.012
63. Logan BE, Hamelers B, Rozendal R, Schröder U, Keller J,
Freguia S, Aelterman P, Verstraete W, Rabaey K (2006) Micro-
bial fuel cells: methodology and technology. Environ Sci Tech-
nol 40(17):5181–5192
64. Krewer U, Song Y, Sundmacher K, John V, Lübke R, Matthies
G, Tobiska L (2004) Direct methanol fuel cell (DMFC): analy-
sis of residence time behaviour of anodic flow bed. Chem Eng
Sci 59(1):119–130
65. Kumar S, Ramamurthy T, Subramanian B, Basha A (2008)
Studies on the fluidized bed electrode. Int J Chem React Eng.
https​://doi.org/10.2202/1542-6580.1610
66. Kreysa G, Pionteck S, Heitz E (1975) Comparative investiga-
tions of packed and fluidized bed electrodes with non-conduct-
ing and conducting particles. J Appl Electrochem 5(4):305–312
67. Rennie AJ, Martins VL, Smith RM, Hall PJ (2016) Influ-
ence of particle size distribution on the performance of ionic
liquid-based electrochemical double layer capacitors. Sci Rep
6:22062
68. Crittenden JC, Trussell RR, Hand DW, Howe KJ, Tchobanoglous
G (2012) MWH’s water treatment: principles and design. Wiley,
New York
69. Fradler KR, Kim JR, Boghani HC, Dinsdale RM, Guwy AJ,
Premier GC (2014) The effect of internal capacitance on power
quality and energy efficiency in a tubular microbial fuel cell.
Process Biochem 49(6):973–980
70. Aguirre-Sierra A, Reija A, Berná A, Salas JJ, Esteve-Núñez
(2014) A Microbial Electrochemical Constructed Wetlands
(METlands): design and operation conditions for enhancing the
removal of pollutants in real urban wastewater. In: Conference:
2nd European Meeting of the International Society for Microbial
Electrochemistry and Technology (EU-ISMET 2014)
71. Austin G, Yu K (2016) Constructed wetlands and sustainable
development. Routledge, London
72. Iza J (1991) Fluidized bed reactors for anaerobic wastewater
treatment. Water Sci Technol 24(8):109–132
73. Nedovic V, Willaert R (2013) Fundamentals of cell immobilisa-
tion biotechnology, vol 8. Springer, New York
74. Aelterman P, Freguia S, Keller J, Verstraete W, Rabaey K (2008)
The anode potential regulates bacterial activity in microbial fuel
cells. Appl Microbiol Biotechnol 78(3):409–418
75. Carmona-Martinez AA, Harnisch F, Kuhlicke U, Neu TR,
Schroder U (2013) Electron transfer and biofilm formation of
Shewanella putrefaciens as function of anode potential. Bioel-
ectrochemistry 93:23–29. https​://doi.org/10.1016/j.bioel​echem​
.2012.05.002
76. Huang JS, Yang P, Li CM, Guo Y, Lai B, Wang Y, Feng L,
Zhang Y (2015) Effect of nitrite and nitrate concentra-
tions on the performance of AFB-MFC enriched with high-
strength synthetic wastewater. Biotechnol Res Int. https​://doi.
org/10.1155/2015/79839​7
77. Santoro C, Guilizzoni M, Baena JC, Pasaogullari U, Casalegno
A, Li B, Babanova S, Artyushkova K, Atanassov P (2014)
The effects of carbon electrode surface properties on bacteria
attachment and start up time of microbial fuel cells. Carbon
67:128–139
78. Das DM (2017) Fuel cell: a bioelectrochemical system that con-
verts waste to watts. Springer, New York
ChemTexts (2019) 5:4 Page 15 of 15  4
79. Bastian P, Kraus J, Scheichl R, Wheeler M (2013) Simulation of
flow in porous media: applications in energy and environment,
vol 12. Walter de Gruyter, Berlin
80. Huh T (1985) Electrical and electrochemical behavior of fluid-
ized bed electrodes. University of California, Berkeley
81. Sabacky B, Evans J (1977) The electrical conductivity of flu-
idized bed electrodes—its significance and some experimental
measurements. Metall Trans B 8(1):5–13
82. Villasenor J, Capilla P, Rodrigo M, Canizares P, Fernandez F
(2013) Operation of a horizontal subsurface flow constructed
wetland–microbial fuel cell treating wastewater under different
organic loading rates. Water Res 47(17):6731–6738
83. Larrosa-Guerrero A, Scott K, Head I, Mateo F, Ginesta A, God-
inez C (2010) Effect of temperature on the performance of micro-
bial fuel cells. Fuel 89(12):3985–3994
84. Behera M, Murthy S, Ghangrekar M (2011) Effect of operating
temperature on performance of microbial fuel cell. Water Sci
Technol 64(4):917–922
85. Michie IS, Kim JR, Dinsdale RM, Guwy AJ, Premier GC (2014)
The influence of anodic helical design on fluid flow and bioelec-
trochemical performance. Bioresour Technol 165:13–20
86. Erable B, Etcheverry L, Bergel A (2011) From microbial fuel cell
(MFC) to microbial electrochemical snorkel (MES): maximiz-
ing chemical oxygen demand (COD) removal from wastewater.
Biofouling 27(3):319–326
87. Xie X, Criddle C, Cui Y (2015) Design and fabrication of bio-
electrodes for microbial bioelectrochemical systems. Energy
Environ Sci 8(12):3418–3441
88. Dennis PG, Virdis B, Vanwonterghem I, Hassan A, Hugenholtz
P, Tyson GW, Rabaey K (2016) Anode potential influences the
structure and function of anodic electrode and electrolyte-asso-
ciated microbiomes. Sci Rep 6:39114. https​://doi.org/10.1038/
srep3​9114
89. Torres CI, Krajmalnik-Brown R, Parameswaran P, Marcus AK,
Wanger G, Gorby YA, Rittmann BE (2009) Selecting anode-
respiring bacteria based on anode potential: phylogenetic, elec-
trochemical, and microscopic characterization. Environ Sci Tech-
nol 43(24):9519–9524. https​://doi.org/10.1021/es902​165y
90. Levar CE, Hoffman CL, Dunshee AJ, Toner BM, Bond DR
(2017) Redox potential as a master variable controlling path-
ways of metal reduction by Geobacter sulfurreducens. ISME J
11(3):741–752. https​://doi.org/10.1038/ismej​.2016.146
91. Peng L, You S-J, Wang J-Y (2010) Electrode potential regu-
lates cytochrome accumulation on Shewanella oneidensis cell
surface and the consequence to bioelectrocatalytic current gen-
eration. Biosens Bioelectron 25(11):2530–2533. https​://doi.
org/10.1016/j.bios.2010.03.039
92. Yoho RA, Popat SC, Torres CI (2014) Dynamic potential-
dependent electron transport pathway shifts in anode biofilms
of Geobacter sulfurreducens. ChemSusChem 7(12):3413–3419.
https​://doi.org/10.1002/cssc.20140​2589
93. Levar CE, Chan CH, Mehta-Kolte MG, Bond DR (2014) An
inner membrane cytochrome required only for reduction of
high redox potential extracellular electron acceptors. MBio
5(6):e02034. https​://doi.org/10.1128/mBio.02034​-14
94. Koch C, Muller S, Harms H, Harnisch F (2014) Microbiomes
in bioenergy production: from analysis to management. Curr
Opin Biotechnol 27:65–72. https​: //doi.org/10.1016/j.copbi​
o.2013.11.006
95. Fricke K, Harnisch F, Schröder U (2008) On the use of cyclic
voltammetry for the study of anodic electron transfer in micro-
bial fuel cells. Energy Environ Sci 1 (1). https​://doi.org/10.1039/
b8023​63h
96. Rodrigo Quejigo J, Rosa LFM, Harnisch F (2018) Electro-
chemical characterization of bed electrodes using voltammetry
of single granules. Electrochem Commun 90:78–82. https​://doi.
org/10.1016/j.eleco​m.2018.04.009
97. Borsje C, Liu D, Sleutels THJA, Buisman CJN, ter Heijne A
(2016) Performance of single carbon granules as perspective for
larger scale capacitive bioanodes. J Power Sources 325:690–696.
https​://doi.org/10.1016/j.jpows​our.2016.06.092
98. Lovley DR (2011) Reach out and touch someone: potential
impact of DIET (direct interspecies energy transfer) on anaerobic
biogeochemistry, bioremediation, and bioenergy. Rev Environ
Sci Biotechnol 10(2):101–105
99. Donlan RM (2002) Biofilms: microbial life on surfaces. Emerg
Infect Dis 8(9):881
100. Rijnaarts HH, Norde W, Bouwer EJ, Lyklema J, Zehnder AJ
(1993) Bacterial adhesion under static and dynamic conditions.
Appl Environ Microbiol 59(10):3255–3265
101. Rittman BE (1982) The effect of shear stress on biofilm loss rate.
Biotechnol Bioeng 24(2):501–506
102. Hamilton M (2003) The biofilm laboratory: step-by-step proto-
cols for experimental design, analysis, and data interpretation.
Cytergy
103. Lewandowski Z, Beyenal H (2013) Fundamentals of biofilm
research, 2nd edn. CRC Press, Boca Raton, FL
104. Pawley J (2010) Handbook of biological confocal microscopy.
Springer, New York
105. Sun D, Cheng S, Wang A, Li F, Logan BE, Cen K (2015) Tem-
poral-spatial changes in viabilities and electrochemical properties
of anode biofilms. Environ Sci Technol 49(8):5227–5235
106. Kramer J, Soukiazian S, Mahoney S, Hicks-Garner J (2012)
Microbial fuel cell biofilm characterization with thermogravi-
metric analysis on bare and polyethyleneimine surface modified
carbon foam anodes. J Power Sources 210:122–128
107. Eaton A, Franson M (2005) Standard methods for the examina-
tion of water and wastewater. American Public Health Associa-
tion, Washington, DC
108. Bhatt P, Kumar MS, Mudliar S, Chakrabarti T (2007) Biodeg-
radation of chlorinated compounds—a review. Crit Rev Environ
Sci Technol 37(2):165–198
109. Aulenta F, Tocca L, Verdini R, Reale P, Majone M (2011)
Dechlorination of trichloroethene in a continuous-flow bioelec-
trochemical reactor: effect of cathode potential on rate, selec-
tivity, and electron transfer mechanisms. Environ Sci Technol
45(19):8444–8451
110. Strycharz SM, Gannon SM, Boles AR, Franks AE, Nevin KP,
Lovley DR (2010) Reductive dechlorination of 2-chlorophenol
by Anaeromyxobacter dehalogenans with an electrode serving
as the electron donor. Environ Microbiol Rep 2(2):289–294
111. Matsuno Y, Tsutsumi A, Yoshida K (1996) Electrode perfor-
mance of fixed and fluidized bed electrodes for a molten carbon-
ate fuel cell anode. Int J Hydrog Energy 21(8):663–671
112. Logan B, Cheng S, Watson V, Estadt G (2007) Graphite fiber
brush anodes for increased power production in air-cathode
microbial. Fuel Cell Environ Sci Technol 41(9):3341–3346
113. Xie X, Ye M, Hu L, Liu N, McDonough JR, Chen W, Alsha-
reef HN, Criddle CS, Cui Y (2012) Carbon nanotube-coated
macroporous sponge for microbial fuel cell electrodes. Energy
Environ Sci 5:5265–5270
Publisher’s Note Springer Nature remains neutral with regard to
jurisdictional claims in published maps and institutional affiliations.
13