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Author(s): Reena Meshram1, Shailesh Kumar Jadhav2



    S.o.S. in Biotechnology, Pt. Ravishankar Shukla University, Raipur (C.G.) 492010, India

Published In:   Volume - 1,      Issue - 1,     Year - 2019

Cite this article:
Reena Meshram and Shailesh Kumar Jadhav (2019) Production of Bioelectricity using Microbial Fuel Cells Fed with Synthetically Designed Wastewater. NewBioWorld A Journal of Alumni Association of Biotechnology, 1(1): 9-12.

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NewBioWorld A Journal of Alumni Association of Biotechnology (2019) 1(1):9-12


Production of Bioelectricity using Microbial Fuel Cells Fed with Synthetically Designed Wastewater

Reena Meshram and Shailesh Kumar Jadhav*

S.o.S. in Biotechnology, Pt. Ravishankar Shukla University, Raipur (C.G.) 492010, India.

*Email- +917712262583




Article history:


18 August 2017

Received in revised form

2 August 2018


4 November 2018


Microbial fuel cells (MFCs) are the electrochemical systems that harness electron from the reduction of organic compounds using microbes as a catalyst. 3 combinations from 4 electrodes that are, Zn (14.9cm×4.9cm), Carbon (14cm×1.5cm), Cu (14.9cm×4.9cm) and Al (14cm×4.5cm) were assessed . Zn-C, as an anode-cathode combination produced maximum voltage of 1.1±0.03V and current 1.5±0.12mA. In present study,gram negative non-fermentative staphylococcus bacterium was isolated from a mediator-less microbial fuel cell, fed with rice bran oil refinery wastewater operated in fed-batch manner. The isolate produced potential of 1.01±0.01V and current of 1.24±0.03mA using synthetic wastewater. The newly isolated bacterium has potential of generating electricity in MFC system and may hold many possibilities with different wastewater as well as in practical applications.



Membrane-less MFC





The demand for energy is surging and the major source for energy is fossil fuel. All the non-renewable resources including fossil fuel are limited and pose negative effect on environment. Concerns about these raise in demand and environmental issues, intense search for renewable, sustainable and green alternatives to fossil fuel are growing (Logan 2004). Bio energy, derived from renewable resources provide novel alternatives to these nature depletive resources such as bio fuel (ethanol and biodiesel)and bio power (pyrolysis and fuel cells) etc. one of the most efficient approaches is Microbial fuel cell (MFC). MFC are the electrochemical systems that use biocatalysts (microorganisms) for the conversion of chemical energy of organic matter to electrical energy under much wider environmental conditions (Allen and Bennetto 1993; Logan 2004).MFC converts chemical energy directly into electricity with variety of substrates, thus is more efficient.

A typical MFC consists of conventional electrochemical system i.e. anode and cathode electrode provided with anaerobic anode and open air cathode. Electrons move to the cathode compartment through the external circuit, and protons migrate through a proton exchange membrane (PEM) or agar salt bridge (Min and Logan 2004; Logan et al. 2006). In cathode compartment, protons get combined with oxygen to oxidize into water. Since the electrons are generated by microbial metabolism, the microbes are of primary importance. Thus, many studies till the date have been made to search for an efficient microorganism like Shewanella putrefaciens (Kim et al. 1999), Geobacter sulfurreducens (Bond  and  Lovley 2003), Rhodoferax ferrireducens (Chaudhuri  and  Lovley 2003)  and  Klebsiella pneumoniae  (Zhang  et  al.  2009).

Rapid industrialization leads to generation of ample amount of wastes, which contaminates the receiving water bodies. The world is facing crucial challenges of its handling, treatment and disposal. The MFC technology offers advantages over the conventional wastewater treatment approaches while generating electricity. It also reduces the amount of sludge and cost of wastewater treatment (Ahn and Logan 2010). The literature shows that in MFCs, various wastewaters have been treated such as sewage (Liu et al. 2004; Ahn and Logan 2010), swine wastewater (Kim et al. 2008), effluent from paper and pulp industry (Huang and Logan 2008), brewery wastewater (Feng et al. 2008), effluent from sugar industry (Abhilasha and Sharma 2009), phenolic wastewater (Luoa et al. 2009), rice mill wastewater (Beheraet al. 2010) and distillery (Mohanakrishna et al. 2010). The present work aims to study isolation of bacterial strain from rice bran oil refinery wastewater and the feasibility of bioelectricity generation by the bacteria using it along with different combination of electrodes in mediator-less double chambered MFC.

Materials and methods

Sample collection

Wastewater from a local oil refinery, Shree Sita Refiners Pvt. Ltd.  Arasnara, Durg, Chhattisgarh (India) has been used for the isolation studies. The wastewater was collected in 5 L sterile airtight plastic containers and was stored at 4±10C for short term. Designed synthetic wastewater was used to check the potential of isolated bacterial strain.

Construction of Microbial Fuel Cell

Dual-chambered H-shape MFC was fabricated with non-reactive, microwave proof polyvinyl chloride containers with a working volume of 750 mL. It comprises of an anode and an open air-cathode chambers. These chambers were kept intact with the aid of adhesive material that is M-Seal and a UPVC pipe of dimension 6.1cm × 1.3 cm was used to physically separate them. The pipes contained agar salt bridge (sodium chloride, 10% and agar, 5%) which acts as a proton exchange material (Momoh and Naeyor 2010; Kumar et al. 2012) (Fig. 1).The MFC were sterilized with saturated ethanol followed by heat sterilization at 850C for 2hr and irradiated with UV for 30 min (Kuashik and Jadhav 2017) and was operated under anaerobic microenvironment.

Figure 1: Schematic diagram of typical two chambered MFC

Three combinations from four electrodes viz C (14cm×1.5cm), Zn (14.9cm×4.9cm), Cu (14.9cm×4.9cm) and Al (14cm×4.5cm) (B. R. Instrument and equipments, Chhattisgarh, India) were checked assuming that larger surface area of electrode may provide larger area for biofilm formation and good electron transfer (Prabowo et al. 2016). External copper wires along with alligator clips were used to connect the electrodes to the digital multimeter (KUSAM-MECO 603) (Logan 2005). The anode compartments were subjected to a leak proof sealing of joints and the exposed metal surfaces sealed with a nonconductive epoxy to avoid corrosion (Kumar et al. 2012). The open air-cathodechamber was filled with 50mM phosphate buffer (pH 7). In present study, oxygen was employed as the final electron acceptor (Feng et al. 2008).

MFC Operation

The MFC setup was run in fed-batch mode. The performance of all the MFCs was evaluated by measuring open circuit voltage (OCV) and current. Constant voltage output was considered as indicator of stable performance of MFC. For the isolation and electrode optimization study, anodic chamber was filled with 750 ml of collected wastewater sample and cathode chamber was filled with phosphate buffer. For screening of electrogenic property of isolated bacterial strain, synthetically designed wastewater (glucose 3.0g/l, FeCl3 25×10-3g/l, NH4Cl 0.5g/l, CaCl2 5.0×10-3g/l, K2HPO4 0.25 g/l, KH2PO4 0.25g/l, MgCl2 0.3g/l, NiSO4 16.0×10-3g/l, CuCl2 10.5×10-3 g L-1, ZnCl2 11.5×10-3g/l, MnCl2 15.0×10-3g/l) and CoCl2 25.0×10-3g/l, with pH 7 was used (Aldrovandi et al. 2009). Anode and cathode were connected to external wiring to complete the circuit. Voltage and current were measured via multimeter. The electrolytic solution was exposed to oxygen present in ambient air for the reduction reaction to occur (Feng et al. 2008; Vignesh and Rani 2012). In anode compartment, bacterium oxidizes fuel, resulting in production of electrons and protons (Gil et al. 2003; Gregory et al. 2004). The external circuit applied for electrons to travel across it and protons get transferred to cathode compartment through salt bridge. In anode compartment anaerobic micro-environment was maintained throughout the operation (Lovely et al. 1993). All the MFCs were operated at ambient room temperature (32 ± 20C).

Statistical Analysis

Voltage and current were recorded in the open circuit using auto-range digital multimeter (KUSAM-MECO 603), after each 1hour time interval for three and five consecutive days, for electrode optimization and isolation studies respectively. All experiments were performed in triplicate. Descriptive analysis of data and calculations were done by SPSS16 and typical values are presented.

Results and Discussion

Optimization of electrode

One of the major challenges associated with MFCs is the cost of the electrode material. The present study elucidates the feasibility of cheaper and easily available materials as an alternative to expensive and sophisticated electrode material (Kim et al. 1999; Logan 2008; Singh and Songera 2012). 3 combinations from 4 electrodes that are Zn-C, Zn- Cu and Zn-Al were checked. (Tab.1). Zn-C electrode combination gave the maximum voltage 1.1±0.02 V and current of 1.5±0.12 mA. The low voltage and current were recorded for the Zn-Al (0.196±0.070V and 0.311 ±0.147mA).

Table 1: Microbial fuel cell performance with rice bran oil refinery industry wastewater using three different pair of electrode combinations.





Voltage (V)

Current (mA)

Voltage (V)

Current (mA)

Voltage (V)

Current (mA)



















Each setup was repeated 3 times and typical values are presented as mean±SE.

Isolation of bacteria from anode biofilm

The bacterial strains were isolated from rice bran oil refinery industry wastewater. Wastewater was serially diluted using distilled water to make series from 10-1 to 10-9. The dilutions were inoculated on plates containing nutrient agar medium and incubated for 48 h. Different mixed bacterial colonies were observed. Bacterial colonies were pure cultured by streak plate method and on the basis of morphology different types of bacterial strain were observed. The isolates were characterized on the basis of cultural and physiological characteristics. The isolates were named as WRS-1 to WRS-7 (in which W stands for wastewater, R for rice industries and S for the name of the industry from where wastewater was collected. Present communication reports about one of the isolate WRS-1. The isolate WRS-1 is found to be gram negative staphylococcus bacterium and was further subjected to various biochemical characterizations (Fig. 2 and Tab. 2).

Table 2: Biochemical characterization of newly isolated bacterium WRS-1.

Biochemical Test


Lactose Fermentative  Test


Dextrose Fermentative Test


Sucrose Fermentative Test


Triple Sugar Iron Agar Test (H2S Production)


Starch Hydrolysis


Urease Test


Methyl Red






Citrate Utilization


Catalase Test



Figure 2: Positive biochemical testes performed by newly isolated bacterium WRS-1

Bioelectricity production by bacterial isolate WRS-1

The isolate WRS-1 produced near about 1.105±0.006V and current of 1.252± 0.005mA in mediator-less MFCs using synthetically designed wastewater (Figure 03). This pattern might be due to high organic load and easy availability range of organic material in synthetic wastewater, for growth of bacterial isolate.

Figure 3: Bioelectricity production from bacterial isolate WRS-1 in mediator-less MFC fed with synthetically designed wastewater

In an overall mechanism, the bacterium utilizes organic components of wastewater for their metabolic activities and generates redox-active molecules that lead to shuttling of electrons between reduced and oxidized compounds. In anode chamber, oxidation of fuel takes place which results in generation of electrons (Bond and Lovley 2003). These electrons are further transferred to Zn anode causing the oxidation of Zn (Zn is converted to Zn2+)when electrons are discharged and transferred through the external wires (Meshram and Jadhav2017). The electrons pass across the external load to reach the cathode chamber. Meanwhile, ambient air is employed as oxygen source at the cathode chamber where final reduction reaction occurs. Oxygen gets electrochemically reduced due to movement of H+ ion from anode to cathode through the proton exchange material and ultimately forming water on the cathode side (Kaushik and Jadhav2017).

This is assumed that transfer of electron takes place via physical contact between bacterial cell membrane or a membrane organelle with the fuel cell anode, because no external redox mediators have been applied to the electrochemical system that could help in accomplishing the electron transfer between the cell and anode. It is reported that release of electrochemically active redox enzymes specifically present in Cytochromes of outer membrane of intact bacterial cells allow transfer of electrons to external solid electron acceptor in MFC (Prasad et al. 2007; Bond and Lovley2003).


Microbial fuel cells (MFCs) are promising electrochemical systems for direct harvesting of electron from range of organic compounds. Zn-C electrode combination as anode and cathode is effective for electricity production. In present study, gram negative non-fermentative staphylococcus bacterium was isolated which was found to be electrogenic. The isolate may also be used in many practical applications such as clearance of organically loaded wastewater from different industrial effluents.

Conflict of interest

Authors had no conflict of interest.


Authors are thankful to the University Grants Commission, New Delhi (India) for providing financial support to the scholar under NET-Junior Research Fellowship scheme; and the Department of Science and Technology, New Delhi for financial support through DIST-FIST scheme to support to School of Studies in Biotechnology, Pt. Ravishankar Shukla University, Raipur (C.G.)


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