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A comprehensive review on emerging constructed wetland coupled microbial

fuel cell technology: potential applications and challenges

Supriya Gupta, Pratiksha Srivastava, Sunil A. Patil, Asheesh Kumar Yadav

PII: S0960-8524(20)31650-3
DOI: https://doi.org/10.1016/j.biortech.2020.124376
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A comprehensive review on emerging constructed wetland coupled microbial fuel

cell technology: potential applications and challenges

Supriya Gupta1,2, Pratiksha Srivastava3, Sunil A. Patil4, Asheesh Kumar Yadav1*

1CSIR-Institute of Minerals and Materials Technology, Bhubaneswar, 751013, Odisha, India
2Academy of Scientific and Innovative Research (AcSIR), CSIR-Human Resource Development Centre, (CSIR-

HRDC) Campus, Ghaziabad, India

3Australian Maritime College, College of Sciences and Engineering, University of Tasmania, Launceston,

Australia, 7248
4Department of Earth and Environmental Sciences, Indian Institute of Science Education and Research Mohali

(IISER Mohali), Knowledge City, Sector 81, SAS Nagar, 140306, Punjab, India

*Corresponding Author, Tel. +91-674- 237-9540; Fax: +91 674 2581160,

Email: [email protected], [email protected]

Abstract

Constructed wetlands (CWs) integrated with bioelectrochemical systems (BESs) are being

intensively researched with the names like constructed wetland-microbial fuel cell (CW-MFC),

electro-wetlands,electroactive wetlands, and microbial electrochemical technologies-based

constructed wetland since the last decade. The implantation of BES in CW facilitates the tuning

of redox activities and electron flow balance in aerobic and anaerobic zones in the CW bed

matrix, thereby alleviating the limitation associated with electron acceptor availability and

increasing its operational controllability. The benefits of CW-BES include high treatment

efficiency, electricity generation, and recalcitrant pollutant abatement. This article presents

CW-BES technology’s journey since its emergence to date, encompassing all the research done

so far, including the basic principle and functioning, bio-electrocatalysts as its machinery,

influential factors for microbial interactions, and operational parameters controlling different

processes. A few key challenges and improvements are also discussed in particular for

electricity harvesting from the systems.

1
Keywords: Constructed wetland-microbial fuel cell, Electroactive wetland, Electro-wetland,

Microbial electrochemical technologies, Microbial Fuel Cell

1. Introduction

The possibility of a humanitarian crisis associated with the availability of freshwater and

energy sources is imminent due to surge in water and energy demand. The shortage in

freshwater availability and rise in energy consumption by the next decade are anticipated to be

40% and 36%, respectively, which demand sustainable solutions to address these problems

concomitantly (Reddy et al., 2019; Jingyu et al., 2020). The existing wastewater treatment

facilities suffer from an imbalance in the work-energy ratio to meet the effluent discharge

standards (Yang et al., 2018; Yuan & He, 2015). It is now recognised that wastewater is a

renewable resource of energy withholding several times higher chemical energy than

demanded by its treatment (Pandey et al., 2016). Thus, the concepts of transforming waste-to-

energy and the development of less energy-intensive wastewater management technologies

have evolved and been widely explored worldwide. The development of cost-efficient and

energy-neutral technologies is currently the most desired approach(Reddy et al., 2019).

Microbial fuel cell (MFC) technology is one of the most promising wastewater treatment

technologies in water-energy nexus (Santoro et al., 2019). It is one of the first

bioelectrochemical systems (BESs) proposed by M.C. Potter (Potter, 1911) utilizing microbes

as biocatalysts for electron generation through anaerobic oxidation of energy-rich organics.

However, its upscaling is challenging majorly due to the requirement of sophisticated and

expensive reactor components and designs contributing to its capital expense (Zhu & Logan,

2013; Behera et al., 2010; Liu et al., 2014). The distinctive characteristic of MFC makes it the

core of several integrated wastewater treatment technologies such as benthic MFC, plant-MFC,

soil-sediment MFC, membrane bioreactor-MFC, desalination-MFC, and hydroponics-MFC

(Srivastava et al., 2020c; Wang & Ren, 2013; Xu et al., 2018b; Xu et al., 2015; Yadav et al.,

2
2020). In the last decades, considerable progress has also been made to integrate MFC into

constructed wetlands (CW) to overcome the challenges associated with individual

technologies.

CW is a cost-effective and green engineered wastewater treatment facility, which operates

passively under the effect of gravitational force. It proficiently treats wastewater from various

origins, such as industrial, municipal, agricultural, storm wastewater, and run-offs (Gupta et

al., 2020a; Vymazal, 2009; Wu et al., 2014). However, the treatment efficiency remains

moderate for organics and inefficient for nutrients removal (Garcia et al., 2010; Gupta et al.,

2019; Srivastava et al., 2019). The main reason is the unavailability of suitable terminal

electron acceptors such as oxygen in a major fraction of the treatment bed, which results in

large land footprint for effective treatment performance (Gupta et al., 2020c; Srivastava et al.,

2019; Yadav et al., 2018). Several approaches that are considered to improve the performance

of CW include design and operational modifications such as circular flow towery hybrid (Peng

et al., 2012), and baffled sub-surface flow system (Wang et al., 2012), effluent recirculation

(Lian-sheng et al., 2006), artificial aeration (Nivala et al., 2013), tidal operation (Wu et al.,

2011), bioaugmentation (Merlin & Cottin, 2012), electron donor supplementation (Zhai et al.,

2013); and integration with other technologies for improving electron acceptor condition (Wu

et al., 2014; Yadav, 2010; Yadav et al., 2018). Feature-wise, the CW bed consists of segregated

upper aerobic and lower anaerobic zones similar to aerobic and anaerobic chambers of MFC.

This structural similarity between CW and MFC makes them compatible for integration (Figure

1). The merger of these two different technologies is called Constructed Wetland-Microbial

Fuel Cell (CW-MFC), representing the new paradigm at water-energy-land nexus. The motive

of innovative integration has been upscaling the features and addressing the limitations of the

individual CW and MFC technologies. Insert Figure 1 here.

3
Integrated CW-MFC is a decade old technology and still in its infancy stage of development

and implementation. Several research studies have been conducted to optimize its efficiency,

both as a treatment plant and an electricity-generating system. Reviews are available on

specific aspects of CW-MFC such as the potential of their integration, principles of microbial

electrochemical processes in CWs, different architectural and operational parameters, and its

challenges and future outcomes (Corbella & Puigagut, 2016; Doherty et al., 2015b;

Guadarrama‐Pérez et al., 2019; Ramírez-Vargas et al., 2018b; Wang et al., 2020b). Recently,

Xu et al. (2019) briefly reviewed the potential environmental applications of CW-MFC other

than electricity generation. Srivastava et al. (2020b) reviewed the electron transfer mechanisms

for pollutant abatement in CW-MFCs. In this review, we encompass the journey of CW-MFC

from the first proof of concept study to the present status and discuss different aspects. These

include efforts to understand the effects of underlying bio-electrochemical and bio-

physiochemical processes for better treatment and electricity output, the interplay of

wastewater treatment and electricity generation processes on the overall performance, and an

overview of advantages, challenges, troubleshooting and potential futuristic applications.

2. Development and Functioning of CW-MFC

CW and MFC are the structural and functional units of CW-MFC. Structurally, CW is an

engineered conglomeration of macrophytes, sandstones or gravels and gradually developed

microbial film in a shallow lined basin with contaminated water flowing through it and

undergoing treatment (Corbella & Puigagut, 2016; Vymazal, 2005; Vymazal, 2007). The depth

of the CW retards air diffusion, and so anaerobic environment prevails at the bottom zone.

However, towards the surface, atmospheric oxygen diffusion and rhizospheric oxygen leakage

create an aerobic regime (Gorgoglione & Torretta, 2018; Brix, 1994). The implantation of

anode and cathode electrodes, which are integral to MFC, in stratified aerobic (high redox

potential, Eh) and anaerobic zones (low redox potential, Eh) in CW (Figure 1) led to the

4
development of CW-MFC technology (Yadav, 2010; Yadav et al., 2012). Besides improved

wastewater treatment, the controllability of Eh is useful for electricity generation in these

systems (Doherty et al., 2015a; Yadav et al., 2012). Overall, CW-MFC configuration provides

naturally occurring conditions for enhanced wastewater treatment and electricity generation.

Other types of integrations, such as snorkel, have also been reported during the last few years.

It is the simplest and practical innovation in the BES that allows the resistance-free movement

of electrons from the anode to the cathode and is solely dedicated to regulated and intensified

treatment performance rather than energy harvest. These have been named as Microbial

Electrochemical Wetland (METland), Electro-wetland, electroactive wetlands (Aguirre-Sierra

et al., 2016; Ramírez-Vargas et al., 2019; Srivastava et al., 2020b) depending on a short-circuit

operational mode. Electro-wetland is designed as a biofilter with complete replacement of stone

or gravel filler substrate with conductive material to form short-circuited CW. The system

limits the voltage generation to zero and maximizes the current flow across the conductive

material filled treatment bed to the maximum attainable value, ensuring the highest possible

electron mediated microbial reaction rates (Hoareau et al., 2019). The microbial community

composition and their relative abundance in electro-wetland have shown a drift from

conventional CW with 4.5 folds enhanced COD removal kinetics (Ramírez-Vargas et al.,

2018a).

2.1. Fundamentals of CW-MFC

Self-purification is the inherent property of any natural water-bearing bodies such as lakes and

wetlands. Several redox potential-dependent microbial processes coordinate to support

pollutant elimination through oxidation and/or reduction of various components

simultaneously (Figure 2). Thus, the natural redox process continues to bio-transform the

wastewater components at a slow rate and is limited to low strength treated water quality.

Microbes utilize several electron donors and acceptors present in wastewater for their growth.

5
However, the electron acceptors are usually limited and get exhausted gradually. Such is a

scenario of any CW where, due to its anaerobic conditions, the unavailability of suitable

electron acceptors (oxygen) hinders the pollutant removal performance (Yadav et al., 2018).

Insert Figure 2 here.

However, in CW-MFC, the supplementation of inexhaustible electron acceptors (anode) and

donor (cathode) in the deficient zones tunes the redox chemistry in its vicinity, thereby

influencing the microbial processes (Wang et al., 2016b). The anode respiring electroactive

bacteria (EAB) tend to proliferate over the electrodes’ surface and supplement metabolic

electrons to it, which flow towards a high redox potential electron acceptor through the external

electrical connection (Corbella et al., 2014). The continuous flow of electrons prevents their

accumulation and thereby circumvents creating a highly reductive environment that favours

methanogenic microbes in the anode region (Wang et al., 2019d). The zones much farther to

the anode are relatively less influenced and continue with the unaltered processes. Electrons

become surplus in the high redox potential cathode region, promoting microbially catalysed

electron mediated reduction reactions. The electrons are deciphered to chemical and microbial

species closer to the cathode with relative ease than those much farther from it. The microbial

processes involved in the degradation of pollutants and electron transfer are complex, and thus

their understanding involves insights into microbial electrochemistry (Gupta et al., 2020c;

Ramírez-Vargas et al., 2018b). The external resistance applied across the electrodes regulates

the electron flow, thereby regulating the treatment and power generation efficiency of the

system. The lower resistance eases electron flow towards the cathode and favours microbial

electron respiration over the anode, which enhances removal efficiency (Ramírez-Vargas et al.,

2018b). On the contrary, higher resistance hinders electron flow across the circuit and

maintains a higher potential difference, which enhances power harvest (Corbella & Puigagut,

6
2018; Fang et al., 2018; Tamta et al., 2020). Accordingly, two simultaneous applications of

CW-MFC can be maximised by optimising the system design.

3. State-of-art of the CW-MFC technology

Yadav reported the concept of CW-MFC for the first time in the Proceedings of 12th

International Conference on Wetland Systems for Water Pollution Control (IWA) in 2010,

followed by the first detailed study in 2012 (Yadav et al., 2012). The study was demonstrated

with a vertical CW-MFC where graphite plates were used as the electrodes in gravel packed

vertical columns. The system effectively reduced the azo bond (N=N) of toxic methylene blue

(redox dye), anaerobically, with the electrons generated and liberated during oxidation of

organics, and a maximum power density of 15.73 mW/m2 was harvested simultaneously. The

preliminary investigations were performed to justify the concept and seek insight into the

electrodes’ role in enhancing anaerobic substrate oxidation processes. The cathodic plants’

contribution was also investigated in promoting the oxidation of pollutants through

rhizospheric oxygen leakage and electricity generation by increased oxygen reduction reaction

(ORR). Thereafter, several studies have reported on various aspects of CW-MFCs, as discussed

in subsequent sections.

3.1. Major architectural aspects and operational modes of CW-MFC systems

The placement of MFC components in the core of CW establishes as a single chamber BES,

with no requirement of proton exchange membrane for its functioning. Several inter-dependent

biological, operational, chemical, and electrical factors such as substrate conversion rate,

anodic and cathodic overpotential, internal resistance, chemical oxygen demand (COD)

concentrations in the anode and cathode regimes, pH and temperature, electrode material type,

electrode surface area, inter-electrode distance, have been identified to affect the performance

of CW-MFC (Srivastava et al., 2019). Some of these factors, such as the anodic and cathodic

overpotential, substrate conversion rate, and electrode material type, regulate the redox

7
conditions in CW-MFC and determine its thermodynamically favourable and unfavourable

conditions for the electron flow (Srivastava et al., 2020b).

With the outlook of extracting maximum efficiency while considering the influential factors,

researchers have explored several designs and hydraulic flow patterns since the innovation of

CW-MFC (Figure 3). The hydraulic flow direction is considered to be directly related to

treatment performance and power output. The vertical upflow-CW-MFC (VUF-CW-MFC)

design has been identified as the most optimistic design and has been rigorously investigated

so far. In VUF-CW-MFC design (Figure 3A), the anode is buried deep in the bed matrix, where

it experiences a high flux of degradable organics required for microbial growth and coulomb

generation. Whereas the cathode is placed at the surface, experiencing minimal organics and

maximum oxygen conditions. The remarkably comparable lower anaerobic and upper aerobic

regions developed in such a design maintains sufficient potential difference to facilitate

electricity generation (Doherty et al., 2015c). The considerable inter-electrode distance,

however, challenges the electrical performance. The early studies on CW-MFC were conducted

with a non-conductive CW bed with conductive electrodes embedded in it and by establishing

an electrical connection across the electrodes (Yadav et al., 2012; Zhao et al., 2013). Gradually,

with the view of increasing surface for the attachment of EAB, the larger fraction of conductive

material, i.e., anode and cathode, were introduced as packets with charge collector embedded

into them (Fang et al., 2013; Oon et al., 2015; Srivastava et al., 2015). The modified electrode

arrangement helped reduce the inter-electrode distance, which reduced the overall internal

resistance of CW-MFC (Oon et al., 2015). The reduced inter-electrode distance along with the

reverse hydraulic flow directions in the anode and cathode regions of CW-MFC (Figure 3B)

further reduced the internal resistance by 67% (Doherty et al., 2015a). However, the treatment

efficiency was adversely affected and reduced considerably. Insert Figure 3

here.

8
Furthermore, several CW-MFC designs have been experimented to improve the total nitrogen

(TN) removal. TN removal involves simultaneous nitrification and denitrification. The

downflow vertical CW-MFC with cathodic macrophyte (Figure 3C) experienced strategic

aerobic-anaerobic environments to facilitate the nitrification-denitrification process and utilize

DO and dissolved organic carbon for nitrogen removal (Wang et al., 2017b). Gupta et al. (2020)

developed a multi-stage CW-MFC assisted by an algal cathode and followed by a sand filter

(figure 3J) to achieve simultaneous denitrification-nitrification-denitrification in a single

assembly and achieve complete removal of nitrogen. Similarly, Srivastava et al. (2020e)

developed a hybrid HF-VUF-CW-MFC (horizontal flow followed by vertical upflow

constructed wetland microbial fuel cell) (Figure 3H). The HF-CW-MFC experienced the

nitrification and anaerobic ammonium oxidation for ammonium removal, and VUF-CW-MFC

carried out denitrification in its lower anaerobic zone.

The HSSF-CW is also a popular CW system with water flowing in parallel to the surface of

CW from inlet to the outlet points, and the hydraulic regime maintained few centimetres below

the gravel surface. The presence of macrophytes provides adequate oxygen supply in the upper

regime through continuous leakage. The HSSF-CW-MFC design (Figure 3G) has been

explored to utilize large surface areas (Corbella Clara, 2015; Srivastava et al., 2020a;

Villasenor et al., 2013). Also, a concept of modular CW-MFC has been investigated to address

the problem of limited performance while upscaling the system. It involves the downscaling of

a single large CW-MFC to several smaller units and upscaling a modular system by connecting

the smaller units of CW-MFC (Figure 3K). Improved treatment efficiency and electrical output

have been reported with this design when electrically networked in series and parallel

connections (Tamta et al., 2020). Tamta et al. (2020) have used serially connected stacked CW-

MFC (Figure 3J) to treat high strength wastewater (COD = 3000 mg/l) with varying external

resistances. The study concluded that decrement in external resistance increased COD removal

9
efficiency. Similarly, stacking in the form of tired CW-MFC (Figure 3F) was investigated by

Xu et al. (2017c), and enhanced COD and TN removal was achieved. However, the electrical

performance was adversely affected by the progress of time. To take advantage of

anaerobically generated biological electrons from a large CW-MFC volume by harvesting them

maximally, Tang et al. (2019) embedded four anodes and one cathode in a 30L pilot-scale CW-

MFC system. They tested it based on different connections, aeration, and effluent recirculation

(Figure 3D) to achieve higher power output and wastewater treatment. The study concluded

that a larger activated cathode area and the parallel connection were useful in achieving

efficient performance. Several authors have worked on developing various CW-MFC designs

with cathode modifications to decrease the internal resistance (Corbella et al., 2016; Gupta et

al., 2020b). For instance, realising the limitation of a high ohmic resistance observed in a single

cathode, Xu et al. (2018c) developed CW-MFC with multi-biocathode assembly and studied a

relationship between nitrogen removal and electricity generation (Figure 3E). The enhanced

simultaneous nitrification and denitrification, and thus overall high nitrogen abatement was

achieved. Aguirre-Sierra et al. (2016) developed the short-circuited CW-MFC without

applying the external electrical circuit for electricity harvesting (Figure 3I). The sole objective

of the developed design was to maximize treatment efficiency.

3.2. Effect of operational parameters on electricity generation and wastewater

treatment efficiency of the CW-MFC systems

3.2.1. Organic Loading Rate

The loading rate determines the substrate available to microbes for degradation and metabolic

activities, growth, and electron generation in BES. The presence of electrodes in CW-MFC

enhances the microbial kinetics for the breakdown of organics present in wastewater by

facilitating anodic respiration. Villasenor et al. (2013) investigated the effect of varying surface

organic loading rates (OLRs: 13.9, 31.1 and 61.1 g COD/m2/d) on the treatment efficiency of

10
CW-MFC. They observed that at low OLR, the anode region removed organics satisfactorily

(up to 100%), and the cathode region remained aerobic to carry out ORR. However, at higher

OLRs, the efficiency decreased to 80-85% with a decrease in cathodic oxygen concentration

and oxidation-reduction potential, which hampered the inter-electrode (anode-cathode)

electron flow. The average and maximum voltage recorded were 700 mV and 1161 mV, and

the maximum current and power densities were 1.22mA/m2 and 0.15mW/m2, respectively.

Srivastava et al. (2020a) reported similar findings in which a closed circuit HSSF-CW-MFC

sustained to perform satisfactorily (98-99%) at low volumetric OLR (0.15 and 0.30 kg

COD/m3/d); however, the performance declined to 95.4% at a high volumetric OLR (0.52 kg

COD/m3/d). Moreover, a noticeable performance improvement of 37.7% was noted for the

closedcircuit design compared to an open circuit HSSF-CW-MFC at a high volumetric OLR

of 0.52 kg COD/m3/d. The study achieved maximum power and current densities of 11.67

mW/m3 and 17.15 mA/m3, respectively. Aguirre-Sierra et al. (2016) analysed the COD removal

rates against OLR under the influence of conductive material as a permanent electron acceptor.

Interestingly, the investigation reported that conductive material equipped CW achieved 5-

folds higher COD removal rate than non-conductive CW. The enhancement was speculated to

be due to the complex interaction amongst the biofilter components and the associated

processes, rather than only current flow through the conductive material. The maximum

treatment and electrical efficiency in CW-MFC is dependent on organic loading rate.

Therefore, possible research directions for future studies can be the determination of effect of

volumetric loading rate on the COD removal efficiency of CW-MFC which will be supportive

in determining the long-term stable and efficient performance of the system.

3.2.2. Role of plants and aeration

Plants play a key role in CW-MFC systems since they liberate oxygen and exudates through

their roots into the rhizosphere zone and assist in the occurrence of biogeochemical cycling of

11
various elements. They provide a large surface area for microbial anchorage and influence the

microbial flora associated with the rhizosphere. The dense and fine network of roots has a

filtering effect, which improve the treated water quality (Oon et al., 2017). Their ability of

rhizospheric oxygen leakage termed as radial oxygen loss (ROL), and release of exudates affect

the nitrification and denitrification rates (Vymazal, 2011). The exudates serve as electron

donors for denitrifiers to remove nitrate, whereas oxygen favours ammonium oxidation in the

cathodic region. The uptake of contaminants such as nitrogen, phosphorus, and metals by plants

has been well reported (Brix, 1994; Vymazal, 2011). Recent studies on CW-MFC have put

efforts to evaluate the significance of plants, through ROL compared to intermittent artificial

aeration (IA) as an economic and passive approach for enhancing treatment. Oon et al. (2017)

have stated that ROL's contribution to treatment efficiency was less than IA but was

significantly higher than the non-planted controls. Moreover, ROL had higher environmental

benefits through decreased resource utilization, thriving diverse ecological niche and CO2

sequestration.

Furthermore, ROL satisfactorily develops a sharp redox gradient essential for MFC placement

in CW compared to IA that excessively aerate the system and develop a conducive environment

for aerobic microbes and heterotrophic activity rather than maintaining anaerobic anode and

developing redox gradient (Srivastava et al., 2017). Srivastava et al. (2017) inferred that with

ROL, the degradation rate of organics decreases with an increase in organic load (0.7 g/l to 2.0

g/l glucose); however, with IA, the degradation rate remains high due to heterotrophic activity.

Additionally, it was also mentioned by Liu et al. (2016)that the characteristics and function of

each plant species are different from each other in terms of wastewater treatment efficacy as a

function of ROL. Oon et al. (2017) used different plant species in CW-MFC named Elodea

nuttallii (submerged macrophyte) and Typha latifolia (emergent macrophyte). They reported

that Elodea nuttallii (submerged macrophyte) leaked out more ROL than Typha latifolia

12
(emergent macrophyte) and enhanced nitrification rate by 17%. Furthermore, Oodally et al.

(2019) operated CW-MFC with three South African wetland macrophytes: Cyperus prolifer,

Wachendorfiathyrsiflora, and Phragmites australis. Among others, Cyperus prolifer showed

the highest COD and orthophosphate removal efficiency of 97±1% and 98%, respectively.

Likewise, Saz et al. (2018) conducted experiments with Typha latifolia, Typha angustifolia,

Juncus gerardii, and Carex divisa, and found the highest performance (88% COD removal)

with Typha angustifolia.

The higher DO concentration and oxidation-reduction potential (ORP), stimulated by plants’

ROL, have shown their inevitable role in generating high voltage (Fang et al., 2013; Liu et al.,

2013; Liu et al., 2014). The role of evapotranspiration on voltage generation was monitored by

implementing MFC into pilot-scale CW planted with Phragmites australis.It was found out

that warm days experienced greater voltage fluctuations due to high evapotranspiration than

the cold seasons (Corbella et al., 2016). It has also been stated that different types of plants

impact the voltage generation phenomenon in CW-MFC depending upon the growth rate and

rhizospheric deposition (Yang et al., 2020; Zhou et al., 2018). The additional aeration through

mechanical devices or macrophytes in the cathode zone is beneficial to the overall performance

of CW-MFC. However, macrophytic aeration is a cost-effective alternative to mechanical

aeration sources, albeit with a slightly lower efficacy. Overall, the evaluation of plant species

with relatively high ROL will be effective in optimizing CW-MFC for treatment of recalcitrant

pollutants and electricity generation, economically.

3.2.3. Electrode material type

The biocompatibility of electrode material is crucial for the flow of electrons from microbes to

an electrical circuit in CW-MFC. The wastewater treatment, bioelectricity generation, and

functional stability of CW-MFC are dependent upon the electrode material or conductive

matrix and its surface energy (Srivastava et al., 2020d; Yakar et al., 2018). The microbial

13
electron transfer is dependent on the biocompatibility of the material. The structural and

functional properties of any material govern the microbial affinity, surface area and porosity,

and habitat quality (Wang et al., 2016a; Yakar et al., 2018). Materials such as metals have

better electrical conductivity and can be used to harvest maximum electrons from the electrical

circuit. However, they are not feasible for use in CW-MFC due to their high cost and microbial

toxicity (Srivastava et al., 2020d). The carbon-based materials have good biocompatibility and

electrical conductivity. Therefore, carbon-based materials such as graphite, carbon cloth/felt,

and activated charcoal are commonly used as the electrodes in CW-MFC (Liu et al., 2014;

Srivastava et al., 2015). Liu et al. (2014) analysed the efficiency of different low-cost bio-

cathodes in CW-MFC and realised GAC combinedwith stainless steel mesh (SSM), as a

suitable biocathode material with a higher power density of 55.05 mW/m2.The effect of four

different anode materials such as graphite rod (GR), SSM, foam nickel (FN), and carbon fibre

felt (CFF), on microbial community composition was also explored in CW-MFC (Wang et al.,

2016a). The study revealed that FN and CFF anodes were more conducive for Proteobacteria

as indicated by their relative abundance. However, CW-MFC equipped with FN electrode

apparently improved relative abundance of Dechloromonas, a denitrifying and phosphate

accumulating microorganism due to presence of Ni in FN electrode.

Ge et al. (2020) reported that pyrite (FeS2) based CW-MFC considerably enriched Geobacter

and sulphate-reducing bacteria in the mesocosm, which enhanced simultaneous nitrate and

phosphorus removal up to 70.1% and 91.2%, respectively, within 6 h of the contact period.Fe

and S elements present in the pyrite-based CW-MFC facilitated additional autotrophic

denitrification by supplementing electrons for nitrate reduction and anaerobic heterotrophic

denitrification process assisted by organic carbon present as an electron donor. During nitrate

reduction, iron intermediates such as Fe2+, Fe3+, Fe(OH)3 complexed with phosphorus to

precipitate it simultaneously (Ge et al., 2020; Wang et al., 2019e). Srivastava et al. (2020d)

14
demonstrated enhanced removal of hexavalent chromium by 49.2% in graphite granules based

electroactive wetlands compared to gravel-based microcosm due to difference in conductivity

of the two materials. SEM-EDX analysis showed that graphite contained carbon (99.5%),

whereas gravel contained several elements, including Fe and Al. Otherwise, gravels are non-

conductive minerals with no significant role in electron transfer hence low treatment

performance. It was revealed that the graphite matrix had a positive impact on microbial

diversity and richness. Furthermore, Yakar et al. (2018) reported comparatively better

treatment performance of zeolite for COD, ammonium, nitrate, and total phosphate removal,

which was 92.1%, 93.2%, 81.1%, and 96.7%, respectively, in comparison to sand and volcanic

cinder. Material characterization analysis evidenced larger specific surface area, different

framework structure, and more porous composition in zeolite favoured anchorage and EAB

division. The contribution of large surface area and elemental composition of electrode material

was elucidated by comparing graphite and Mn ore anode electrodes for pollutants removal,

antibiotic resistance genes fate and bacterial community evolution (Liu et al., 2019). Mn ore

anode had a higher abundance of EAB and functional genes for electron transfer. The

abundance of methanogens and corresponding functional genes along with several antibiotic

resistance genes were inhibited due to the occurrence of the dissimilatory metal reduction

process. The electrode material is very crucial in governing the microbial community

composition of biofilm formed over it. Several unmodified natural and modified materials have

been explored to analyse the abundance of microbes. However, the available literature so far is

insufficient to get any insight into the processes responsible for selecting the electrode and its

biocompatibility. Thus, further research is required to develop improved understanding of the

selection of electrode materials in CW-MFC.

3.2.4. Electrode positioning

15
Electrode positioning is crucial while considering current output in CW-MFC. The anaerobic

and aerobic conditions at the anodic and cathodic regimes, respectively, are needed for its

efficient functioning. Moreover, maintaining minimum inter-electrode distance is essential for

reducing the internal resistance. Therefore, the anode and cathode electrodes should be placed

considering both requisites for achieving maximum treatment output (Doherty et al., 2015a;

Doherty et al., 2015c). The effect of the distance between the electrodes was studied by Fang

et al. (2017) to optimise the dye and COD removal by CW-MFC. The placement of anode

closer to the bottom and interelectrode gap of 13.2 cm achieved maximum COD and dye

removal of 71.4% and 91.05%, respectively. The smaller gap of 6.6 cm had even higher COD

removal (89.89%); however, dye removal decreased. Xu et al. (2017a) analysed the placement

of anodes and cathodes in CW-MFC with different variations, as bottom anode-rhizosphere

cathode CW-MFC (BA-RC-CW-MFC), rhizosphere anode-air cathode CW-MFC (RA-AC-

CW-MFC), and bottom anode-air cathode CW-MFC (BA-AC-CW-MFC). It was noted that

satisfactory COD removal occurred when the anode was placed at the bottom, and the cathode

did not receive much COD. However, when the anode was placed towards the surface in the

macrophyte rhizosphere’s vicinity, the anode could not get enough contact time and volume to

degrade COD satisfactorily and prevent it from reaching the cathode. Regarding the cathode

placement, the air-water interface cathode performed better than the rhizospheric cathode in

RA-AC-CW-MFC and BA-AC-CW-MFC at an increased loading rate from 9.2 g/m2/D to 92

g/m2/D. The results indicated the performance was due to the accumulation of exudates from

plants over the cathode and low oxygen secretion from plants. The system with air cathode

produced higher voltage, whereas the rhizospheric cathode produced zero voltage. However,

for air cathode, the system with anode at the bottom produced 329±16 mV voltage and

maximum power density of 11.21 mW/m2 and, rhizospheric anode produced 73±25 mV and

1.05 mW/m2,under 27.6 gCOD/m2/dOLR. It indicated insufficient organics degradation in

16
rhizospheric anode resulting in the decreased cathodic oxygen concentration and obstructed

cathodic reduction reactions and electron flow. The optimization of electrode position with

respect to the required aerobic and anaerobic environment to reduce the internal resistance are

key points of consideration while keeping higher electricity generation and pollutant removal

goals. Corbella et al. (2016) reported that the placement of cathode 1-2 cm above water level

was its optimum position. Though much attention has been given to this aspect of the CW-

MFC domain by researchers, considerable work has not been done with the volumetric increase

of the system, which is essential for its real field application.

3.2.5. Electrode number

Several studies have reported that a single electrode, anode or cathode, remains insufficient for

the maximum possible harvest of electrons from EAB at the anode or dispensing electrons from

the cathode to the reducible moieties in the vicinity (Oon et al., 2016; Tang et al., 2019; Xu et

al., 2018c). The application of multiple electrodes in the CW-MFC system is directly related

to the increase in the total surface area of electrodes with respect to the total volume of anode

or cathode. Such an arrangement ensures maximum electron transfer to the anode and cathode.

The placement of multiple cathodes at suitable positions modifies the redox potentials in

treatment bed, resulting in decreased energy losses (Xu et al., 2018c). Xu et al. (2018c)

investigated the electricity generation performance of CW-MFC with the application of the

multi-cathode system in contrast to a single cathode. Multi-cathode system exchanged more

electrons and supplemented higher oxygen reduction sites, which showed a potential loss from

97.85 mV to 46.09 mV for the anode and 221.5 mV to 45.89 mV for the cathode. As an

outcome, the energy losses decreased for both the anode and cathode by 51.76 mV and 175.61

mV. Moreover, the power density improved from 12.56 to 26.16mW/m2.

Oon et al. (2016) studied the multi-anode system with variation in organic loading rates 314 ±

9.6 mg/L and 624 ± 16.3 mg/L and observed that the anode at the bottom (at 7 cm from the

17
bottom) alone removed the maximum COD and the remaining anodes (at 21 cm and 36 cm)

received low COD concentration for removal. The decreased COD near upper anodes increased

the DO concentration from 0.49 mg/L to 1.44 mg/L at 21 cm and 36 cm, respectively, favouring

partial ammonium nitrification removing 96% ammonium in the effluent. On the other hand,

denitrification occurred at the anode placed at 7 cm and 21 cm, resulting in 97% nitrate

removal. Tang et al. (2019) investigated the performance of multiple anodes embedded CW-

MFC in series and parallel connection to observe the contribution of connections. The

parallelly connected CW-MFC outperformed the serially connected CW-MFC.Cathode

improvement was also considered simultaneously in the study by aeration and recirculation.

The maximum power density for a system with electrodes in parallel along with aeration,

recirculation, and both aeration-recirculation was 1.55 mW/m2, 3.09 mW/m2, 7.99 mW/m2,

respectively. Thus, it is crucial to increase the anode and cathode’s surface area by splitting

their size and determine the electron exchange capability for enhanced treatment and electrical

performance. Also, the investigation on electrical and hydraulic patterns of connection of

multiple electrodes of a single module or multiple modules could be effective in determining

the electron flow mechanisms, voltage reversal, capacitance behaviour of CW-MFC for

enhancing the power output.

3.2.6. Applied external resistance

The external resistance across the electrodes impacts the exchange current density and so the

pollutant removal performance of CW-MFC (Corbella & Puigagut, 2018; Tamta et al., 2020).

Moreover, it has been stated that due to large size and volume, CW-MFC usually has high

external resistance, which leads to low voltage output (Gajda et al., 2020; Srivastava et al.,

2020a; Srivastava et al., 2020e). Due to the large surface area, the internal resistance of the

system gets impacted by several factors such as the initiation of redox reactions driven by

activation energy, ohmic losses due to external resistance of the system, and concentration

18
losses (Sanchez et al., 2019; Srivastava et al., 2020b). Fang et al. (2017) have stated that in

CW-MFC, the ohmic losses can be governed by the system’s external resistance, and it is

dependent on the distance between the electrodes. Tamta et al. (2020) investigated the effect

of external resistance across the electrodes of serially connected stacks of pilot-scale CW-MFC,

which were distantly placed. The restricted electron flux towards the cathode influenced the

COD and ammonium removal largely. At resistances ranging from 80-1000 Ω, COD removal

varied between 98.5-96.3%; however, further increase to 1500Ω decreased it to 77.6%, which

was a significant drop in performance. Fang et al. (2018) studied the effect of external

resistance on dye removal in up-flow CW-MFC and observed that the electron flow from the

anode at 620Ω provided the highest decolourization efficiency with a maximum current density

of 539 mA/m3. Likewise, Wang et al. (2019c) investigated the role of external resistance on

the reduction of greenhouse gas emissions and observed that external resistance considerably

increased CH4 emission and decreased CO2 emission. Since various factors such as large size

and volume are involved in CW-MFC the operational controllability of external resistance is

not easy to manage; however, there is chance to control the losses inside the system by choosing

eco-friendly highly conductive filling materials, and change in design. Therefore, further

research is required to optimise the role of external resistance and to overcome the problem of

high external resistance of CW-MFC by modifying its internal factors.

3.3. Performance assessment with various types of target pollutants

Many recalcitrant and hazardous pollutants are often present in wastewater, such as dye laden

textile wastewater, antibiotics, and personnel care products (PCPs) laden municipal

wastewater, and pesticide-contaminated agricultural runoff. Recalcitrant pollutants are

resistant to microbial oxidation and thus persistent in nature. The potential of CW-MFC for the

degradation of recalcitrant pollutants has been explored by several researchers (Di et al., 2020;

Fang et al., 2015; Fang et al., 2016; Li et al., 2019; Srivastava et al., 2020d; Wang et al., 2019b;

19
Xie et al., 2018). Recent studies have reported on the trace accumulation of antibiotics in water

systems, which has happened due to the excessive usage of antibiotics and similar products by

mankind. The deposition of antibiotics traces leads to the development and accumulation of

antibiotic resistant bacteria and resistance genes in the environment. In this context, Zhang et

al. (2016) targeted the removal of tetracycline (TC) and sulfamethoxazole (SMX) in CW-MFC

system and revealed an effective reduction in TC and SMX concentration from 400µg/l,

1000µg/l and 1600 µg/l to less than 1.65 µg/l along with the generation of power density 57.8

mW/m2. The electron mediated oxidation of SMX to its reduced metabolites was found

responsible for reducing concentration and encouraged the use of CW-MFC. At the same time,

Zhang et al. (2017) have encouraged to use the external supply of electrons for addressing the

high concentration SMX in wastewater. The applied voltage from 0.6 to 1.2 V enhanced the

removal efficiency of SMX up to 99.92%. Li et al. (2019) shed light on a similar finding by

demonstrating 9.3% and 18% higher ibuprofen (IBP) and bisphenol A (BPA) removal with an

initial concentration of 10 mg L-1 in closed circuit CW-MFC as compared to open circuit, and

the significant difference was attributed to the anode region. It was also revealed that the higher

loading rate and lower HRT increased the microbial toxicity; however, Proteobacteria and

Bacteroidetes increased significantly, showing increased tolerance to toxicity. Nitrobenzene

(NB) is an aromatic compound with NO2 as functional moiety attached to it. Xie et al. (2018)

explored the anaerobic reduction process of NB in CW-MFC, and the findings highlighted that

the electron mediated process in CW-MFC achieved 83.48% and 78.3% nitrobenzene (NB)

and COD removal, respectively, as compared to 73.96% and 70.46% in CW; and 81.54% and

74.13% in MFC alone, respectively.

Similarly, Di et al. (2020) analysed the ROL’s role by investigating the presence of three

different wetland plant species, namely Scirpus Validus, Typha orientalis, and Iris pseudacorusin

CW-MFCfor the removal of NB ranging from 20-200mg/l. Scirpus Validus showed maximum

20
tolerance to NB at the highest concentration of 200mg/l. It liberated relatively higher root oxygen

(DO = 2.57 ± 0.17 mg/L) and accumulated high biomass (16.42 ± 0.18 g/m2), which resulted in a

higher power density of 19.5 mW/m2 and NB removal efficiency of 93.9%. Furthermore,

Srivastava et al. (2020d) have demonstrated chromium (Cr(VI)) removal in electroactive wetlands

and achieved 99.9% of Cr(VI) removal in graphite-based CW. It was 42.9% higher than gravel-

based CW. However, the electrical output is not well achieved, requiring more studies to dictate

the role of recalcitrant pollutants on the system’s treatment and electrical performance.

3.4. Electroactive bacteria: The real machinery of CW-MFC

The microbial community composition primarily impacts the wastewater treatment and

electricity generation performance of CW-MFC (Wang et al., 2016a). The metabolic functions

of microorganisms, determined by their functional genes, have a crucial role in electron transfer

and pollutant removal (Xu et al., 2018a). EAB's dominance is widely accepted and reported in

CW-MFC as they serve as a key component for the enhanced performance. Their symbiotic

association with non-EABs has also been reported in BES. However, their relative abundance

increases or decreases according to the immediate environment surrounding them (Cheng &

Logan, 2011; Srivastava et al., 2020e; Wang et al., 2019a). The replacement of the wetland’s

non-conductive filter matrix with electroconductive material stimulates EAB’s growth and

abundance by encouraging the extracellular electron transfer (EET) phenomenon (Srivastava

et al., 2020d). Generally, EAB feeds on simple carbon sources such as acetate for the generation

of electricity. Therefore, depending upon the complexity of the carbon source, they form a

consortium and syntrophic association with several kinds of non-EAB that hydrolyse or

ferment the complex organics to simpler forms such as acetate (Kiely et al., 2011; Wang et al.,

2020a).

EAB's respiration using the external electron acceptor, either soluble or insoluble, depends on

the potential difference between the electron transport components of microbes and the electron

acceptor. The microbes tend to interact with the acceptor that yields higher energy to them

21
(Srivastava et al., 2020b). Until now, two types of mechanisms have been proposed for

microbial interaction with external electron acceptors, namely direct or indirect transfer

mechanisms (Logan, 2009). As the microbes grow over the electrode surface, they form a

multi-layered biofilm over it. The monolayer of biofilm in direct contact with the electrode

typically utilise outer-membrane redox proteins and cytochrome cascades for direct electron

transfer to the electrode (Logan, 2009). However, the bacterial layers away from the electrode

andare unable to contact it directly develop nanowire structures to connect with the electrodes

or other bacteria via extracellular conductive matrix in order to transfer electrons, which is

referred to as a direct interspecies electron transfer (Bonanni et al., 2012; Busalmen et al.,

2008). On the contrary, in indirect electron transfer mechanism, soluble electron shuttles or

mediators are used for extracellular electron transfer (Arends & Verstraete, 2012; Ramírez-

Vargas et al., 2018a). The role of redox mediators or shuttles can be explored for enhancing

electron transfer rate and wastewater treatment in CW-MFC.

The reported predominant bacterial phyla in CW-MFCs are Proteobacteria, Firmicutes,

Bacteroidetes, Acidobacteria, Actinobacteria, Chloroflexi, Lentisphaerae, and Spirochaetes

(Aguirre-Sierra et al., 2016; Corbella Clara, 2015; Li et al., 2019; Li et al., 2016; Lu et al.,

2015; Ramírez-Vargas et al., 2018a; Rathour et al., 2019; Srivastava et al., 2020d; Srivastava

et al., 2020e; Wang et al., 2016a; Wang et al., 2017a; Xu et al., 2018a). At class level,

Alphaproteobacteria, Betaproteobacteria, Gammaproteobacteria,Deltaproteobacteria and

Anaerolineae, and at a family level, Rhodocyclaceae, Anaerolineaceae, Bacteroidales,

Desulfovibrionaceae, Spirochaetaceae, Rhodospirillaceae, Desulfobulbaceae, Weeksellaceae,

Oxalobacteraceae, Flavobacteriaceae, Nocardioidaceae, Comamonadaceae,

Xanthomonadaceae, Rikenellaceae, Bacteroidaceae, Rhizobiales,Planctomycetaceae and

Clostridiaceae have been reported to be relatively abundant (Wang et al., 2016a; Wang et al.,

2016b; Wang et al., 2019a; Xu Et al., 2018a). However, to get insight into the microbial

22
functioning with respect to the exposed surrounding the understanding of microorganisms at

the genus level is desirable. Wang et al. (2016b), while working with different pH noted that

alkaline pH favoured the growth of genera Fusibacter, Gemmata, Planctomyces, and

Prosthecobacter; however, the genera Neisseria, Zoogloea, Dechloromonas, and Geobacter

dwelled well under the slightly acidic and neutral conditions in the anodic zone. It was

speculated by Wang et al. (2016a) that the presence of metallic Ni promoted the growth of

Dechloromonas, which are capable of autotrophic denitrification and phosphate-accumulation.

Wang et al. (2019a) revealed the abundance of Nitrospira, Thauera, and Dechloromonas as

dominant nitrifying and denitrifying genera at the anode while treating ammonium-free,

nitrate-free and normal wastewaters. The relatively abundant nitrate-reducing bacteria included

Dechloromonas, Desulfobulbus, Flavobacterium, Propionivibrio, and Geobacter in closed

circuit CW-MFC (Wang et al., 2016b). In some studies, Dechloromonas and Geobacter have

been identified as dominant electricity-generating bacteria in organics fed systems (Corbella et

al., 2015; Wang et al., 2016b). Clostridium andDesulfovibrio have been identified as dye

decolorizing bacteria in the anode region (Rathour et al., 2019). As per the literature, the

abundance of EAB varies at the anode and cathode of CW-MFC, signifying their differing roles

in the two regimes (Liu et al., 2020a). For instance, anode respiring bacteria such

asComamonas are the essential microbes at the anaerobic anode rather than at the cathode and

have been reported to produce electricity when present at the anode (Liu et al., 2020a).

The abundance of Bacillus andThauera, genera, which are denitrifying bacteria, has been

reported in the cathode. Xu et al. (2018a) reported the predominance of Geobacter,

Desulfovibrio, and Bacillus at the cathode. Xu et al. (2019) identified the relative abundance

of Nirtospira in the cathode, which plays a role in nitrite to nitrate oxidation process.

Depending upon the constituents of rhizodeposits from the cathodic plant, different bacteria

such asDesulfobulbus (oxidizes propionate, hydrogen, or lactate), Clostridium (oxidises

23
glucose, starch or xylose), Desulfovibrio (sulfate-reducing bacteria), and

Propionivibriopredominate (Wang et al., 2016a; Wang et al., 2019a; Wang et al.,

2017a).Through direct or indirect electron transfer, these bacteria play a significant role in CW-

MFCs and are responsible for enhancing the overall process. Further understanding of CW-

MFC bacteria and their role in different processes is needed to achieve the desired performance

by these systems.

4. Key challenges of the CW-MFC and its potential futuristic applications

Amongst several proposed applications of CW-MFC, wastewater treatment with concurrent

electricity generation remain its primary application. So far, the CW-MFC technology has

shown improvement in the treatment efficiency of CW. It is mainly ascribed to the anode

influence on anaerobic oxidation of pollutants and cathode influence on the reduction reactions.

The reported studies reveal that MFCs have achieved high coulombic efficiencies and power

densities, ranging from few to several thousand Watts per cubic meter with respect to their

anodic volumes. However, with achieved volumetric power densities in milliWatts range, CW-

MFC has not been able to perform equivalent to MFCs in terms of electrical performance. The

fundamental reason is the much higher volume of subjected wastewater with respect to the

exposed electrode (conductive material) surface area. Therefore, the recovery of electrons

generated in CW-MFC has been lower (Wang et al., 2017c). The increased volume is

accountable for a decrease in the maximum power density by 2-3 orders of magnitude due to

an increase in ohmic losses. The stacked modular CW-MFC holds the potential to add up the

voltage generated by each unit and improve the electricity generation to a value that can operate

the low energy devices. However, stacked modular CW-MFC often suffers from an

unpredictable voltage reversal phenomenon when some of the unit cells work as a power

supplying batteries and others as power-consuming loads (Gajda et al., 2020; Tamta et al.,

2020; Xu et al., 2017c). Thus, energy harvesting is challenging. In such a scenario, several

24
next-generation applications of CW-MFC can be explored, as discussed in the following sub-

sections.

4.1. Biosensors

The regulation and monitoring of water bodies receiving the treated water are essential to

protect the ecology and environment. The commonly used analytical methods are resource-

intensive and time-consuming. The sensor-based online analytical tools developed are much

effective and accurate. The real-time monitoring is helpful in early alarming and taking timely

precautionary and control measures. MFC-based biosensors have been investigated for

estimating biological oxygen demand (BOD) for the last several years (Corbella et al., 2019).

The organics present in the wastewater is metabolized by anodic microbes to generate electric

current establishing a direct and linear relationship between biodegradable organics and current

in MFCs. Biosensing is the indigenous application of MFC apart from electricity harvesting

since current signals are already generated by it. In addition to that, unlike other biosensors, it

does not require a transducer. Generally, microbes are sensitive to the contaminants present in

the environment around them. Any sudden change in the composition of water, for example,

the presence of toxins, or change in organics or nutrient concentration, flowing through the

anodic biofilm acts as a stimulus to them. It can trigger the change in their metabolic pathways

and kinetics, which can be observed as fluctuation in the current generated (Chouler et al.,

2018). Therefore, these biosensors have emerged as simple online analytical tools. Xu et al.

(2017b) demonstrated the first study on CW-MFC as a biosensing tool for COD in the range

of 0-1000mg/l and concluded that for a lower concentration of COD (<100mg/l), the lower

response time (approx. 2 h) is reliable in comparison to concentration >200mg/l that achieve

stable voltage plateau at 5 h. Also, at COD concentrations higher than 700mg/l, the signals are

again very close to each other due to the system’s saturation for substrate utilization. At a range

between 200 to 700mg/l, quite differentiated and reproducible voltage signals were generated.

25
Corbella et al. (2019) stated that the biosensing ability of CW-MFC is a qualitative assessment

tool for continuous monitoring of influent water quality, rather than a precise measurement tool

for COD due to overloading, which causes failure after few weeks. The field-scale application

of CW-MFC for decontamination of groundwater from benzene, methyl-tert-butyl ether, and

ammonium along with its in situ benzene oxidation, was performed by Wei et al. (2015). The

quality assessment was assessed through the linear relationship between current density and

benzene removal. The work demonstrated the generation of electric signals when effluent

contained benzene remnants, whereas the system stopped to generate current as benzene was

completely removed. Nevertheless, the application of CW-MFC as an alarming tool for

monitoring the water quality, and maintaining the treatment system, still needs to be

investigated and optimized for implementation. Thus, there is an enormous scope to investigate

the correlation between the pollutant concentration and electrical output to demonstrate the use

of generated bioelectricity from the system for online monitoring of the pollutants. This scope

would potentially be a sustainable regulating factor for the system’s operation such as,

monitoring system failure or toxicity enhancement in the system.

4.2. Using power management system for CW-MFC

The power management systems (PMS) have come up to drive the electronic loads by power

generated in low energy devices like MFCs and its archetypes (Winfield et al., 2014). PMS is

an electronic setup consisting of a power source like CW-MFC, super-capacitor for charge

storage and dispensing, and DC/DC converter, which boosts the low input voltage generated

by the power source to a high output voltage. The limitation of most of the MFC based systems

is the low input voltage to the capacitor, which governs its charging and discharging limit (Xu

et al., 2018b)., A charge pump was introduced between MFC and super-capacitor to enhance

the limited charge storage, which worked with a low input electrical potential and regulated

charging speed. Another component, transformer, was introduced in PMS setup to replace the

26
charge pump and increase the super-capacitor’s storage capacity. Currently, PMS are widely

used for powering devices. For instance, Xu et al. (2018b) investigated PMS integration with

biocathode CW-MFC to extract more power by harvesting the electrons stored within the anode

capacitor. Three operational strategies were tested to compare the electron harvesting

efficiencies: normal CW-MFC with continuously loaded external resistance (CL),

intermittently loaded external resistance, duty cycling (DC), and intermittently loaded external

resistance with a capacitor, capacitor duty cycling (CDC). The electron harvesting efficiencies

were 76.1%, 84.4%, and 91.16% with CL, DC, and CDC operational modes, respectively. The

two experimental D values, 31.6%, and 20%, supported the statement by obtaining the

enhanced electron harvesting efficiencies as 91.16% and 95.1%, respectively, in CDC mode.

It is the only study reported in CW-MFC with PMS, and further research can give new

directions to its electrical applications. One approach of utilising the step up bi-current

generated by CW-MFC can be its direct utilization of mechanical devices such as aerator,

stirrer, and recirculating pump to cut down the operational capital. Furthermore, the

supplementation of current generated for carrying out ex-situ advanced oxidation processes,

bio-electro-Fenton reactions, and external power supply for supporting bioelectrochemical

systems can be extended.

4.3. Electrocoagulation-driven enhanced treatment processes

Electrocoagulation (EC) is an electrochemical method of in-situ coagulant formation by

electrolysis of a sacrificial anode such as iron (Fe) or aluminium (Al) for the pollutant removal

(Ju et al., 2014). The EC process effectively treats industrial wastewater, which contains

recalcitrant pollutants such as dye and metals. It has also been integrated into CW, and the

removal of several pollutants such as sulfamethoxazole, nitrate, ammonium, phosphate, COD,

and sulphate has been demonstrated (Gao et al., 2017; Ju et al., 2014; Liu et al., 2020b). The

EC integrated CW mainly works on the principle of electrolysis of sacrificial or non-sacrificial

27
 conductive electrodes. In a recent EC-CW study, Liu et al. (2020b) have demonstrated an

electrolysis integrated tidal flow-CW for removing sulfamethoxazole and concluded EC-CW

has a higher potential for wastewater treatment even at full scale. However, for long-term

operation, the choice of sacrificial electrodes needs to be considered. Likewise, Gao et al.

(2017) examined electrolysis integrated horizontal subsurface-flow CW for nitrogen and

phosphorus removal. The study concluded that the electro-dissolution of iron and in

situformation of ferric iron coagulant enhanced the nitrate and phosphorus removal

significantly. These limited studies suggest that the integration of EC in CW has enormous

potential for wastewater treatment; however, future research is recommended to consider its

long-term operation feasibility.

Conclusions

The CW-MFC technology represents an innovative approach for enhancing wastewater

treatment efficiency with simultaneous electricity generation. Among different BES integrated

systems, it is the most practical innovation used to intensify the desired process. So far, various

designs and processrelated aspects of CW-MFC have been investigated at laboratory scales.

However, the technology has still not been studied at the field scale except for the electroactive

wetlands. For upscaling, the CW-MFC technology has several research gaps that need to be

addressed, and optimisation of the technology is required.

Supplementary information:

E-supplementary data for this work can be found in the e-version of this paper online.

Acknowledgements

Authors acknowledge the financial assistance through the research grant of NASF, (ICAR,

New Delhi) (NASF/CA-6031/2017-18) and CSIR, India Grant MLP-37 and MLP-59.

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45
Figures captions:

Figure 1. Schematic representation depicting the similarities in a microbial fuel cell (MFC) and

constructed wetlands (CWs) configurations; and the possibility of implanting MFC inside CW,

leading to the emergence of integrated CW-MFC technology.

Figure 2. (a) Schematic representation depicting the microbially-catalysed processes that can

occur at varying bed depths owing to redox potentials in CW-MFC and CW. Metabolically

produced electrons and electrodes influence the redox gradient in (b) CW-MFC, and (c)

Electro-wetland/ Microbial Electrochemical Snorkel. Eh = Redox potential; OP = Oxidation Potential;

RP = Reduction Potential.

Figure 3: Different designs of CW-MFC systems operated in varying hydraulic flow patterns,

(A) Upflow; (B) Simultaneous upflow-downflow; (C) Downflow; (D) Multi-anode (E) Multi-

cathode; (F) Tiered; (G) Horizontal Subsurface flow; (H) Hybrid horizontal flow-vertical flow;

(I) Short-circuit; (J) Multi-stage; (K) Stacked Modular CW-MFC.

Symbolic Representations: Influent ( ); Effluent ( ); Cathode ( ); Anode (

); Resistance ( ); Membrane ( ); macrophytes (P).

46
Figure 1.

47
Figure 2.

48
Figure 3.

49
 Author’s contribution statements

Roles/Tasks Contributory person’s name

Conception & design of the work Dr. Asheesh Yadav, Supriya Gupta,
Pratiksha Srivastava
Data Collection Supriya Gupta
Data analysis and interpretation Supriya Gupta
Drafting the article Supriya Gupta & Pratiksha Srivastava
Critical revision of the article Dr. Asheesh Yadav & Dr. Sunil Patil
Final approval of the version to be Dr. Asheesh Yadav
published

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Highlights:
 A comprehensive review on the progress of constructed wetland integrated microbial
fuel cell
 Insights into design aspects and operational parameters of CW-MFC
 Associated challenges of CW-MFC and futuristic applications
 Electroactive bacteria as bio electrocatalysts catalysing high removal kinetics

51