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An innovative mixotrophic approach of distillery spent wash with sewage wastewater for biodegradation and bioelectricity generation using microbial fuel cell
Journal of Water Process Engineering 23 (2018) 306–313
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Journal of Water Process Engineering journal homepage: www.elsevier.com/locate/jwpe
An innovative mixotrophic approach of distillery spent wash with sewage wastewater for biodegradation and bioelectricity generation using microbial fuel cell Jagdeep K. Nayak, Amit, Uttam K. Ghosh
T
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Department of Polymer and Process Engineering, Indian Institute of Technology Roorkee, India
A R T I C LE I N FO
A B S T R A C T
Keywords: Distillery spent wash MFC Bioelectricity Scenedesmus abundans
In the present study, H-type microbial fuel cell (H-MFC) has been used to treat distillery spent wash (DSW) diluted with sewage wastewater (SWW) at different mixing ratio. Mixture of DSW and SWW was used as substrate in anaerobic environment. The cathodic compartment was filled with BBM media and microalgae (Scenedesmus abundans). CO2 generated during breakdown of organic substrates by anaerobes present in DSW was utilized by microalga Scenedesmus abundans for photosynthesis. A consortium of two bacteria (Pseudomonas aeruginosa and Bacillus cereus) was used for the 21 days period of DSW treatment. Significant reduction in chemical oxygen demand (COD) in the order of 66% to 78.66% was observed. At 50:50 ratio of DSW and SWW, total dissolved solid (TDS) of 39.66% and total suspended solid (TSS) of 97% were removed. A maximum power density (PD) of 836.81 mW/m2 and open circuit voltage (OCV) of 745.13 mV were obtained along with biomass yield of 0.74 g/L d−1 with above ratio of DSW and SWW. This study demonstrated that proper dilution of Distillery spent wash with sewage wastewater may leads to an efficient wastewater remediation and energy production.
1. Introduction The extensive use of fossil fuels, especially oil and gas, in recent years, has lead to the depletion of fossil fuels, environmental pollution and efficient development of renewable energy sources with higher performance [1–4]. The new alternative sustainable energy technologies are need to be eco-friendly, energy neutral and efficient [5]. Microbial fuel cells (MFCs), a hybrid biochemical device, have been tested as a promising technique for wastewater treatment with energy recovery by many researchers [6–8]. Microorganisms are the mediator to convert a large variety of biodegradable organic compounds into CO2, water and energy [5,9]. This facilitates microbial interaction to convert chemical energy to electrical energy and production of value added products through the metabolic activity of microorganisms [2,10–12]. A general design of a two chambered MFC consists of anodic compartment where microorganisms help in oxidative conversion of the substrate. Simultaneous chemical reductive conversion takes place in cathodic compartment. The electrons produced in anodic chamber, pass through the external circuit and at the same time protons passing through a proton exchange membrane (PEM) react with an oxidizing agent, such as oxygen, at the cathode surface which leads to close the
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Corresponding author. E-mail address: [email protected] (U.K. Ghosh).
https://doi.org/10.1016/j.jwpe.2018.04.003 Received 18 January 2018; Received in revised form 22 March 2018; Accepted 2 April 2018 2214-7144/ © 2018 Elsevier Ltd. All rights reserved.
circuit [13–16,37]. With integration of multiple process, elements and experimental conditions, the setup may also have positive influence on the overall wastewater treatment efficiency. The conversion efficiency of organic wastes to bio-energy and economic viability depend on the sources of waste, its chemical composition, characteristics and concentration of the components that can be converted into bio-energy [17]. Distillery spent wash has become a major source of environmental pollution due to the presence of high organic load, dark brown color and unpleasant odor in it. It also contains considerable nutrients in terms of potassium, sulphur, nitrogen and phosphorus as well as large amount of micronutrients like Ca, Cu, Mn, and Zn [18]. Among all the conventional treatment processes, anaerobic treatment is widely accepted practice and also it has been tried at pilot and full scale operations [19]. Prior research has been carried out on the treatment of distillery spent wash in microbial fuel cells (MFCs) to utilize the high organic matters as an oxidizing agent for production of electricity. Brewery wastewater containing COD of 2000 mg/L was treated in dual chambered (rectangular) MFCs (0.2 L anode volume) to obtain power density of 305 mW/m2 at 30 ± 2 °C with 80% removal of COD [20]. Treatment
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Fig 1. Schematic diagram of MFCs used in this study.
2.2. Wastewater collection and characterization
of high strength alcohol distillery spent wash in an integrated AFBMFCs system showed 80–90% removal of COD and a maximum power density of 124.03 mW/m2 was attained with an external resistance of 120 Ω [21]. Traditional aerobic or anaerobic wastewater treatment technologies are energy consuming with emission of large amount of carbon dioxide (CO2) as the major gaseous end product when either glucose or acetate was used as substrate [22]. As algae are recognized as excellent CO2 sinkers and oxygen is generated simultaneously in their metabolic activities, several researchers showed interest to use algae in cathodic chamber for oxygen production which acts as a terminal electron acceptor [22–24]. Wang [22] reported a maximum voltage output of 706 ± 21 mV (1000 Ω) when Chlorella vulgaris was grown in cathodic chamber. In this study, sewage wastewater was mixed in a different proportion with distillery spent wash to examine the wastewater treatment, bioelectricity generation as well as utilization of CO2 for biomass production using dual chambered MFC. Four different concentration of distillery effluent mixed with sewage were used as substrate. Microalgae Scenedesmus abundans was grown in cathode chamber. CO2 produced in the anode chamber was supplied to the cathode for the use of microalgae for their metabolism and growth. The overall performance of the MFC was evaluated in terms of COD removal, metallic cations removal, maximum voltage production, maximum power density and biomass production along with specific growth rate and doubling time.
The raw distillery spent wash (DSW) was collected from the biogas production unit of ethanol distillation plant and the sewage wastewater (SWW) was collected from domestic sewage treatment plant. Both the samples were preserved at 4 °C prior using. Heavy metals were characterized using inductively coupled plasma optical emission spectrometry (ICP-OES) (Teledyne Leeman Labs, Prodigy SPEC, USA). COD, TDS, TSS and TS were determined using method provided by APHA (1992). Electrical conductivity and pH was measured using digital meter (HI 3512, Hanna Instruments). Samples were prepared by mixing distillery spent wash and sewage wastewater in the ratios of 100:0, 75:25, 50:50 and 25:75 by v/v and were denoted as DSW100 + SWW0, DSW75 +SWW25, DSW50 + SWW50, DSW25 + SWW75, respectively. 2.3. Experimental setup An ‘H’ type dual chambered microbial fuel cell (Fig. 1) was designed and fabricated (Fig. 2) using transparent Plexiglas material of thickness 0.5 cm which consists two equal volume cylindrical chambers (each with working volume of 900 mL) separated by proton exchange membrane (PEM). The height and diameter of each chamber were 15 cm and 10 cm, respectively. Graphite rod with 17 cm length and 1.2 cm diameter were used as electrodes in both the chambers. The electrodes were connected to an external resistor of 100 Ω in a loop configuration and the voltage generated was monitored by multi-meter (Mashtech India pvt.ltd.). In this experiment pretreated Nafion-117 from DuPont was used as proton exchange membrane. Pretreatment of nafion-117 membrane was done by dipping the membrane first in a solution of H2O2 (3% (v/v) for 1 h at 80 °C and then in H2SO4 (0.5 M) for 1 h at 80 °C. The membrane was washed with boiling water after each solution treatment [25]. Pretreated (autoclaved) wastewater sample was fed in batch mode for 21 days in the MFC at 25 ± 1 °C. Anaerobic condition was maintained in anodic chamber by purging nitrogen gas was for around 15 mins. The gasses produced in anodic chamber including CO2 were transferred to cathodic chamber through an outlet duct. The gas was utilized by microalgae in cathodic chamber. BBM medium was used in the cathodic chamber for the growth of Scenedesmus abundans. Cathodic chamber was kept under temperature controlled environment and illumination using 4 LED strips with light intensity of 94.6 μmol m−2 s−1. The light and dark cycle maintained was 16 h: 8 h. A sample of 2 mL of sample was collected every day for test of different parameter test.
2. Materials and methods 2.1. Culture collection and cultivation Active bacterial cultures of Pseudomonas aeruginosa (MCC No. 2622) and Bacillus cereus (MCC No. 2240) were procured from National Center for Cell Science, Pune, India. Both the bacterial cultures were cultivated in nutrient agar with sodium chloride media for 48 h at 25 ± 1 °C in incubator. After incubation period of 48 h, cultures were transferred in a single media to develop an efficient bacterial consortium. Scenedesmus abundans (NCIM No. 2897) was obtained from National chemical Laboratory, Pune, India and cultivated in BBM medium with the following components per liter: NaNO3, 0.25 g; MgSO4. 7H2O, 0.075 g; NaCl, 0.025 g; : K2HPO4, 0.075 g; KH2PO4, 0.175 g; CaCl2 2H2O, 0.025 g; H3BO3, 0.011 g; EDTA, 0.05 g; KOH, 0.031 g; FeSO4·7H2O 0.0049 g; H2SO4, 0.001 mL and trace elements (ZnSO4·7H2O, 0.00882 g MnCl2·4H2O 0.00144 g, MoO3, 0.0071 CuSO4·5H2O, 0.00157 g Co (NO3)2·6H2O 0.00049 g). The microalga Scenedesmus abundans was cultivated in 1L Erlenmeyer flasks with 750 mL of autoclaved BBM medium at 25 ± 1 °C under continues illumination (22W, LED tube light) with light intensity of 94.6 μ mol m−2 s−1. There was no supply of CO2 externally except the availability in the atmosphere.
2.4. Electrochemical and chemical analysis Open circuit voltage was monitored and recorded at regular interval of 1hr for 21 days by digital multi-meter connected to 100 Ω external 307
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Fig. 2. Lab scale image of MFCs used in this study.
resistances (R) in series. Power was calculated by using Ohm’s formula, P = V2/R where V and R are the voltage and resistance respectively. The power was normalized to the anode surface in order to calculate power density (mW/A2). Power density was calculated based on the electrode surface area as, PD = P/A where A is the total surface area of the electrode in m2. The COD removal efficiency (%) was calculated using the following equation:
COD Removal (%) =
Ci − Cf Ci
× 100
(1)
Where Ci, Cf are the initial and final COD respectively. 2.5. Algal growth Fig. 3. Percentage removal of COD during the incubation period of 21 days. DW50 + SWW50 showing highest removal of 78.6%.
The concentration of algal biomass was collected in every day basis and was measured by UV spectrophotometer (Shimadzu Corp, Japan) at 680 nm. For estimation of dry biomass by gravimetric process, the sample was collected every day from the algal cultivation chamber, centrifuged at 6200 rpm for 5 min and vacuum dried overnight at 104 °C. Biomass productivity and specific growth rate were calculated as follows:
in cell voltage over time indicates the capability of the inoculated consortia of culture to remove COD of the distillery spent wash and generate electricity. The value of the voltage generated was decreased after a steady removal efficiency of COD was attained due to unavailability of organic nutrient after 17 days. In case of 100:0 v/v sample, lowest removal efficiency of COD (66%) was observed due to the presence of high organic load in the distillery wastewater as it was very difficult to sustain microbes in high COD environment. However a longer lag phase of more than 10 days was observed for higher dilution of DSW was observed. The efficiency of COD removal is increased (78.6%) at 50:50 (v/v) due to less complexity of organic nutrient however a little decreased of organic removed efficiency (66%) was noticed in case of 25: 75 (v/v) due to non-availability of nutrients. Mixed culture as used in the present study showed better COD removal efficiency of 78.6% as compared to single strain treatment (72.84%) dairy wastewater as reported by Venkata Mohan [26]. COD values are considered as an index for survival of living organism in wastewater [27]. Lesser activity of electrogenic consortia and decrease in mobility of charge transfer due to ohmic resistance under such higher concentration of feed might be the reason of reduced current generation. The gradual decrease in efficiency of the MFC might have happened due to overloading of the substrate.
Biomass mass productivity = (final dry biomass − initial dry biomass)/ cultivation period (2) Specific growth rate (μ) = [ln(X1) − ln(X0)]/t1 − t0
(3)
Where X1 and X0 are biomass concentrations at t1 and t0 days respectively. Doubling Time = ln (2)/μ
(4)
3. Result and discussion 3.1. Wastewater treatment 3.1.1. COD removal efficiency The COD concentration in wastewater is an important factor which limits the power generation tendency. The dual chambered ‘H’ type MFC was operated for 21 days in batch mode using four different dilutions of wastewater. The correlation between power density calculated from voltage outputs and COD concentration over the entire incubation period was recorded as shown in Fig. 4. The dilution of distillery effluent with sewage wastewater (50:50 v/v) showed highest electricity generation than the other diluted distillery sample. COD removal efficiency of four different samples varied in between 66% to 78.66%. The steady incline in COD reduction percentage with increase
3.1.2. Pollutant removal efficiency The consumption of macronutrients and micronutrients by the microorganism to enhance its growth and hence reduction of the nutrient concentration in the wastewater supports the purpose of the advanced wastewater treatment. The various physicochemical parameters as well as 308
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Fig. 4. Relation between COD removal (%) and Power density (PD) during the incubation period at different concentration A) DSW100 + SWW0, B) DSW75 + SWW25, C) DSW50 + SWW50, D) DSW25 + SWW75 of distillery spent.
complex of the metal ions inside the cell and later reduce the toxic metal ions to a non-toxic state. Among the bacterial community many research showed that bacillus genus has maximum biosorbents capacity of inorganic pollutant.
macro and micro nutrients were analyzed, before treatment of different samples as recorded in Table 1. In this study the maximum reduction percentage of TDS and TSS of the four samples DSW100 + SWW0, DSW75 + SWW25, DSW50 + SWW50, DSW25 + SWW75 were found 27, 32, 39.66, 32.66 and 64.66, 88, 97, 94, respectively as shown in Fig. 7. The TDS and TSS levels directly related to the various parameters such as EC, HRT etc. In the present study, the maximum reductions of heavy metals like Mg, P, Cd, Ca, Cu, Mn, Pb in different sample were 60.68%, 58.12%, 77.45%, 62.78%, 95.63% and 96.29%, respectively as shown in Fig. 9. Trace metals like K+, Na+, Mg2+, Mn2+, Cu2+and Ca2+, respectively play a crucial role in bacterial growth because of their major participation in the activation of various metabolic pathways like energy consumption and storage. These mechanisms of removal of heavy metals include the efflux of metal ions outside the cell, accumulation and forms
3.2. Production of electricity Four MFCs were used in this current study were fed with distillery effluent and sewage wastewater under different concentration. All the MFCs were operated with proper anaerobic condition maintained in the anodic chamber for 21 days. The output data of voltage was monitored with multi-meter and illustrated in Fig. 5. Stable voltage growth was seen after 12–14 days of incubation periods, which indicates the highest degradation of organic substances after this period. Increasing the
Table 1 Physiochemical parameters. PARAMETERS
DISTILLERY SPENT WASH (DSW)
SWW(mg/L)
DW75 + SWW25
DW50 + SWW50
DW25 + SWW75
Color Odor pH COD (mg/L) TDS (mg/L) TSS (mg/L) TS (mg/L) Total Alkalinity (mg/L) Calcium (mg/L) Total Phosphorous (mg/L) Total Potassium (mg/L)
Dark brown Unpleasant 4.5 ± 1.5 65000–75000 37500 6500 44000 2160 1850 14680 1000
Light brown Unpleasant 7 ± 0.5 870 554 410 964 450 62 5 4
Light brown Unpleasant 6.5 ± 1 (Adjusted) 59000 28300 5000 33300 1970 1420 11050 775
Light brown Unpleasant 6.5 ± 1(Adjusted) 36750 19800 3700 23500 1350 980 7350 565
Brown Unpleasant 6.5 ± 1(Adjusted) 18950 9800 1975 11780 890 510 3690 300
309
310
63.5 ± 1.5 98−62 60.3–85.7 72.84 64.8 ± 0.3 92 – 78.6 412 ± 12 – – 354.29 323.4 ± 4 130 645 (mW/m3) – 202 ± 6 50.6–67.1 151.2–283.5 124.35 123.5 ± 3 930 457 (mW/m3) 836.81 Nafion PVAA/SSA Nafion-117 Nafion-117 Nafion-117 Clayware ceramic Nafion Nafion-117 Lysinibacillussphaericus L. sphaericus SN-2 Aspergillus awamori Pseudomonas aeruginosa, Bacillus cereus Graphite Plate Carbon cloth Carbon cloth Graphite plate Graphite plate Carbon felt graphite fiber brush Graphite rod Dual Chambered Air cathode Dual chambered Single chambered Dual chambered Dual chambered Dual chambered Dual chambered
(−) denote not present.
Microorganism Wastewater Electrode
Fig. 6. Power density (PD) in different concentration of distillery spent wash. Maximum PD of 836.81 mW/m2 in DSW50 + SWW50.
Type of MFC
Table 2 Comparative study of COD removal and Electricity production with current study.
PEM
wastewater COD concentration, increased the conductivity of the solutions which might help to improve the performance of MFC [38] but extended start-up period of 8–9 days might be due to the availability of higher concentration of organic matter for metabolic reaction in distillery effluent and inappropriate growth environment for biocatalyst [28]. The output of 100% DSW also showed higher power density from than other previous studies. The peak power density and COD removal efficiency was observed as 836.81 mW/m2 and 78.66%, respectively for sample DSW50 + SW50 at optimum environment as shown in Fig. 3. The maximum voltage generation of different dilution of DSW100 + SWW0, DSW75 + SW25, DSW50 + SW50 and DSW25 + SWW75 were 346.2 mV, 414.6 mV, 745.13 mV 615.1 mV and, respectively. The mixture of DSW and SWW were utilized as an organic substrate for bacterial biocatalyst and produces electrons (e−) and protons (H+) through redox reaction which facilitating power generation Venkata Mohan et al. [26]. After 10 days of incubation period, the MFC yielded a relatively stable operating potential of 193.33 mV in 100% distillery effluent. The generation of maximum voltage and power density of 346.2 mV and 180.42 mW/m2, respectively, noticed on 17th day and the output was higher than previous studies. From Figs. 5 and 6, it can be seen that, for 50% diluted distillery effluent the voltage generation and power density were increased up to 745.13 mV and 836.81 mW/m2, respectively, on 16th day. In 25% distillery and 75% sewage treated wastewater the maximum voltage of 615.1 mV and power density of 570.23 mW/m2 on 17th day were registered. For distillery effluent diluted with SWW the output was found higher as compared to the previous studies with distillery effluents using MFCs (Table 2). This decrease of power
Distillery Distillery Alcohol Distillery Distillery Distillery post methanation distillery effluent (PMDE) Distillery
References COD Removal (%) Current density (mA/m2) Power density (mW/m2)
Fig. 5. Open circuit voltage generation of different dilution ratio distillery spent wash. Maximum OCV of 745.13 mV in DSW50 + SWW50.
[29] [30] [31] [32] [33] [28] [34] This study
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generation is due to the lower concentration of organic substances which leads the effective electron (e−) discharge. 3.3. Biomass production In general, growth of the microalgae depends upon the cultivation media, culture condition and optimized environment. In this study, the culture conditions are same but in the MFCs CO2 for algae growth was supplied from the anode chamber to the cathode chamber in which algae was cultured. It is shown in principle that gaseous CO2 generated from substrate degradation in anode chamber can be completely removed and converted into algal biomass in the cathode chamber. Furthermore, CH4 was reported not to be the main gaseous product in MFCs. Scenedesmus abundans showed the maximum biomass yield of 0.74 g/L and biomass productivity 0.038 g/Ld−1 in 50% dilution of distillery spent wash over 21 days of cultivation period as shown in Fig. 8, which indicates the maximum anaerobic digestion in anodic chamber with supply of CO2 to cathodic chamber. From Fig. 8, the almost same growth pattern was found in all the different concentrations of wastewater. Scenedesmus abundans has a lag phase of 5 days before going to the exponential phase. It attained the stationary phase between 16 and 21 days of cultivation period. Previous studies [35] showed, a promising biomass production by Scenedesmus abundans.
Fig. 8. Biomass growth (g/L) of Scenedesmus abundans in cathodic chamber for all four sample. Maximum growth of 0.74 g/Ld−1 of DSW50 + SWW50.
dioxide source with BBM medium is the most suitable for Scenedesmus abundans as used in the present work. 4. Conclusions
3.3.1. Specific growth rate and doubling time Specific growth rate and doubling time are the major parameters of algal proliferation which indicate the relationship between the adaptation and growth in a particular condition. Table 3 indicates that the specific growth rate (0.20 d−1) and doubling time (3.38) of algae Scenedesmus abundans with the combination of sample DSW50 + SWW50 is consistently higher than all the other three samples with short of doubling time. Al-shatri et al. [36] reported specific growth rate of 0.104 d−1at 25 °C for growth of Scenedesmus dimorph on in BBM medium under constant photon flux density. In the present study all the four experiments showed higher specific growth rate in comparison to the data reported by Ali Hussain for Scenedesmus abundans. Thus the combination of anaerobic treated DSW50 + SWW50 sample as carbon
In this study, the efficiency of H-type two chamber MFC using mixture of distillery wastewater and sewage wastewater as a substrate was systematically observed. The feasibility of power generation using microalgae (Scenedesmus abundans) was demonstrated. The maximum power density of (836 mW/m2) and maximum OCV of (745.13 mV) in this study were found relatively higher as compared to other reports. The production of CO2 by the degradation of organic and inorganic compounds could be used by microalgae to promote their growth as well as oxygen generation at cathode. The maximum growth of biomass 0.74 g/Ld−1 was noticed. Due to dilution of highly organic distillery wastewater with sewage wastewater and effective adaptation of both
Fig. 7. Removal (%) of TSS and TDS in four different concentration of distillery spent wash.
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Fig. 9. Microbial degradation (%) of metallic anion and micronutrients using MFC for incubation periods of 21 days. Table 3 Biomass, Specific growth rate, doubling time and Biomass Productivity of Scenedesmus abundans. Sample
Final Biomass (g L−1)
Initial Biomass (g L−1)
Specific Growth Rate (d−1)
Doubling Time
Biomass Productivity (g/L d−1)
DSW100+ SWW0 DSW75 + SW25 DSW50 + SW50 DSW75 + SWW25
0.51 0.58 0.74 0.43
0.01 0.02 0.01 0.01
0.18 0.16 0.20 0.17
3.70 4.32 3.38 3.87
0.027 0.028 0.038 0.021
bacteria and algae in both chambers separately might be the reason of higher efficiency of MFC. This study suggests that proper dilution of distillery wastewater and optimized condition for microalgae growth can be a promising way for higher power generation in MFC operation. Acknowledgement The first author would like to thank the Ministry of Human Resources and Development (MHRD), Government of India for providing financial support to successfully carry out this research work. References [1] A. Elmekawy, H.M. Hegab, K. Vanbroekhoven, D. Pant, Techno-productive potential of photosynthetic microbial fuel cells through different configurations, Renew. Sustain. Energy Rev. 39 (2014) 617–627, http://dx.doi.org/10.1016/j.rser.2014. 07.116. [2] Z. Du, H. Li, T. Gu, A state of the art review on microbial fuel cells: a promising technology for wastewater treatment and bioenergy, Biotechnol. Adv. 25 (2007) 464–482, http://dx.doi.org/10.1016/j.biotechadv.2007.05.004. [3] W. Miran, M. Nawaz, J. Jang, D.S. Lee, Sustainable electricity generation by biodegradation of low-cost lemon peel biomass in a dual chamber microbial fuel cell, Int. Biodeterior. Biodegrad. 106 (2016) 75–79, http://dx.doi.org/10.1016/j.ibiod. 2015.10.009. [4] K. Samal, R.R. Dash, P. Bhunia, Performance assessment of a Canna indica assisted vermifilter for synthetic dairy wastewater treatment, Process. Saf. Environ. Protect. 111 (2017) 363–374. [5] P. Pandey, V.N. Shinde, R.L. Deopurkar, S.P. Kale, S.A. Patil, D. Pant, Recent advances in the use of different substrates in microbial fuel cells toward wastewater treatment and simultaneous energy recovery, Appl. Energy 168 (2016) 706–723, http://dx.doi.org/10.1016/j.apenergy.2016.01.056. [6] B.E.B. Logan, Microbial Fuel Cells, (2008), http://dx.doi.org/10.1016/B978-0-44453199-5.00098-1. [7] D.R. Lovley, Microbial fuel cells: novel microbial physiologies and engineering approaches, Curr. Opin. Biotechnol. 17 (2006) 327–332, http://dx.doi.org/10. 1016/j.copbio.2006.04.006. [8] K. Rabaey, W. Verstraete, Microbial fuel cells: novel biotechnology for energy generation, Trends Biotechnol. 23 (2005) 291–298, http://dx.doi.org/10.1016/j. tibtech.2005.04.008.
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Scenedesmus abundans: a potential feedstock for high quality biodiesel production, Bioresour. Technol. 156 (2014) 42–47, http://dx.doi.org/10.1016/j.biortech.2013. 12.127. [36] A.H.A. Al-shatri, E. Ali, N.K.N. Al-shorgani, M.S. Kalil, Growth of Scenedesmus dimorphus in different algal media and pH profile due to secreted metabolites, Afr. J. Biotechnol. 13 (2014) 1714–1720, http://dx.doi.org/10.5897/AJB2013.13455. [37] K. Samal, R.R. Dash, P. Bhunia, Effect of hydraulic loading rate and pollutants degradation kinetics in two stage hybrid macrophyte assisted vermifiltration system, J. Biochem. Eng. 132 (2018) 47–59. [38] Y. Feng, X. Wang, B.E. Logan, H. Lee, Brewery wastewater treatment using aircathode microbial fuel cells, Appl. Microbiol. Biotech. 78 (2008) 873–880.
of distillery wastewater in microbial fuel cell facilitating decolorization and desalination along with power generation, J. Hazard. Mater. 177 (2010) 487–494, http://dx.doi.org/10.1016/j.jhazmat.2009.12.059. [33] N. Samsudeen, T.K. Radhakrishnan, M. Matheswaran, Effect of isolated bacterial strains from distillery wastewater on power generation in microbial fuel cell, Process Biochem. 51 (2016) 1876–1884, http://dx.doi.org/10.1016/j.procbio. 2016.06.007. [34] A. Kaushik, A. Chetal, Power generation in microbial fuel cell fed with post methanation distillery effluent as a function of pH microenvironment, Bioresour. Technol. 147 (2013) 77–83, http://dx.doi.org/10.1016/j.biortech.2013.08.004. [35] S.K. Mandotra, P. Kumar, M.R. Suseela, P.W. Ramteke, Fresh water green microalga
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Journal of Water Process Engineering journal homepage: www.elsevier.com/locate/jwpe
An innovative mixotrophic approach of distillery spent wash with sewage wastewater for biodegradation and bioelectricity generation using microbial fuel cell Jagdeep K. Nayak, Amit, Uttam K. Ghosh
T
⁎
Department of Polymer and Process Engineering, Indian Institute of Technology Roorkee, India
A R T I C LE I N FO
A B S T R A C T
Keywords: Distillery spent wash MFC Bioelectricity Scenedesmus abundans
In the present study, H-type microbial fuel cell (H-MFC) has been used to treat distillery spent wash (DSW) diluted with sewage wastewater (SWW) at different mixing ratio. Mixture of DSW and SWW was used as substrate in anaerobic environment. The cathodic compartment was filled with BBM media and microalgae (Scenedesmus abundans). CO2 generated during breakdown of organic substrates by anaerobes present in DSW was utilized by microalga Scenedesmus abundans for photosynthesis. A consortium of two bacteria (Pseudomonas aeruginosa and Bacillus cereus) was used for the 21 days period of DSW treatment. Significant reduction in chemical oxygen demand (COD) in the order of 66% to 78.66% was observed. At 50:50 ratio of DSW and SWW, total dissolved solid (TDS) of 39.66% and total suspended solid (TSS) of 97% were removed. A maximum power density (PD) of 836.81 mW/m2 and open circuit voltage (OCV) of 745.13 mV were obtained along with biomass yield of 0.74 g/L d−1 with above ratio of DSW and SWW. This study demonstrated that proper dilution of Distillery spent wash with sewage wastewater may leads to an efficient wastewater remediation and energy production.
1. Introduction The extensive use of fossil fuels, especially oil and gas, in recent years, has lead to the depletion of fossil fuels, environmental pollution and efficient development of renewable energy sources with higher performance [1–4]. The new alternative sustainable energy technologies are need to be eco-friendly, energy neutral and efficient [5]. Microbial fuel cells (MFCs), a hybrid biochemical device, have been tested as a promising technique for wastewater treatment with energy recovery by many researchers [6–8]. Microorganisms are the mediator to convert a large variety of biodegradable organic compounds into CO2, water and energy [5,9]. This facilitates microbial interaction to convert chemical energy to electrical energy and production of value added products through the metabolic activity of microorganisms [2,10–12]. A general design of a two chambered MFC consists of anodic compartment where microorganisms help in oxidative conversion of the substrate. Simultaneous chemical reductive conversion takes place in cathodic compartment. The electrons produced in anodic chamber, pass through the external circuit and at the same time protons passing through a proton exchange membrane (PEM) react with an oxidizing agent, such as oxygen, at the cathode surface which leads to close the
⁎
Corresponding author. E-mail address: [email protected] (U.K. Ghosh).
https://doi.org/10.1016/j.jwpe.2018.04.003 Received 18 January 2018; Received in revised form 22 March 2018; Accepted 2 April 2018 2214-7144/ © 2018 Elsevier Ltd. All rights reserved.
circuit [13–16,37]. With integration of multiple process, elements and experimental conditions, the setup may also have positive influence on the overall wastewater treatment efficiency. The conversion efficiency of organic wastes to bio-energy and economic viability depend on the sources of waste, its chemical composition, characteristics and concentration of the components that can be converted into bio-energy [17]. Distillery spent wash has become a major source of environmental pollution due to the presence of high organic load, dark brown color and unpleasant odor in it. It also contains considerable nutrients in terms of potassium, sulphur, nitrogen and phosphorus as well as large amount of micronutrients like Ca, Cu, Mn, and Zn [18]. Among all the conventional treatment processes, anaerobic treatment is widely accepted practice and also it has been tried at pilot and full scale operations [19]. Prior research has been carried out on the treatment of distillery spent wash in microbial fuel cells (MFCs) to utilize the high organic matters as an oxidizing agent for production of electricity. Brewery wastewater containing COD of 2000 mg/L was treated in dual chambered (rectangular) MFCs (0.2 L anode volume) to obtain power density of 305 mW/m2 at 30 ± 2 °C with 80% removal of COD [20]. Treatment
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Fig 1. Schematic diagram of MFCs used in this study.
2.2. Wastewater collection and characterization
of high strength alcohol distillery spent wash in an integrated AFBMFCs system showed 80–90% removal of COD and a maximum power density of 124.03 mW/m2 was attained with an external resistance of 120 Ω [21]. Traditional aerobic or anaerobic wastewater treatment technologies are energy consuming with emission of large amount of carbon dioxide (CO2) as the major gaseous end product when either glucose or acetate was used as substrate [22]. As algae are recognized as excellent CO2 sinkers and oxygen is generated simultaneously in their metabolic activities, several researchers showed interest to use algae in cathodic chamber for oxygen production which acts as a terminal electron acceptor [22–24]. Wang [22] reported a maximum voltage output of 706 ± 21 mV (1000 Ω) when Chlorella vulgaris was grown in cathodic chamber. In this study, sewage wastewater was mixed in a different proportion with distillery spent wash to examine the wastewater treatment, bioelectricity generation as well as utilization of CO2 for biomass production using dual chambered MFC. Four different concentration of distillery effluent mixed with sewage were used as substrate. Microalgae Scenedesmus abundans was grown in cathode chamber. CO2 produced in the anode chamber was supplied to the cathode for the use of microalgae for their metabolism and growth. The overall performance of the MFC was evaluated in terms of COD removal, metallic cations removal, maximum voltage production, maximum power density and biomass production along with specific growth rate and doubling time.
The raw distillery spent wash (DSW) was collected from the biogas production unit of ethanol distillation plant and the sewage wastewater (SWW) was collected from domestic sewage treatment plant. Both the samples were preserved at 4 °C prior using. Heavy metals were characterized using inductively coupled plasma optical emission spectrometry (ICP-OES) (Teledyne Leeman Labs, Prodigy SPEC, USA). COD, TDS, TSS and TS were determined using method provided by APHA (1992). Electrical conductivity and pH was measured using digital meter (HI 3512, Hanna Instruments). Samples were prepared by mixing distillery spent wash and sewage wastewater in the ratios of 100:0, 75:25, 50:50 and 25:75 by v/v and were denoted as DSW100 + SWW0, DSW75 +SWW25, DSW50 + SWW50, DSW25 + SWW75, respectively. 2.3. Experimental setup An ‘H’ type dual chambered microbial fuel cell (Fig. 1) was designed and fabricated (Fig. 2) using transparent Plexiglas material of thickness 0.5 cm which consists two equal volume cylindrical chambers (each with working volume of 900 mL) separated by proton exchange membrane (PEM). The height and diameter of each chamber were 15 cm and 10 cm, respectively. Graphite rod with 17 cm length and 1.2 cm diameter were used as electrodes in both the chambers. The electrodes were connected to an external resistor of 100 Ω in a loop configuration and the voltage generated was monitored by multi-meter (Mashtech India pvt.ltd.). In this experiment pretreated Nafion-117 from DuPont was used as proton exchange membrane. Pretreatment of nafion-117 membrane was done by dipping the membrane first in a solution of H2O2 (3% (v/v) for 1 h at 80 °C and then in H2SO4 (0.5 M) for 1 h at 80 °C. The membrane was washed with boiling water after each solution treatment [25]. Pretreated (autoclaved) wastewater sample was fed in batch mode for 21 days in the MFC at 25 ± 1 °C. Anaerobic condition was maintained in anodic chamber by purging nitrogen gas was for around 15 mins. The gasses produced in anodic chamber including CO2 were transferred to cathodic chamber through an outlet duct. The gas was utilized by microalgae in cathodic chamber. BBM medium was used in the cathodic chamber for the growth of Scenedesmus abundans. Cathodic chamber was kept under temperature controlled environment and illumination using 4 LED strips with light intensity of 94.6 μmol m−2 s−1. The light and dark cycle maintained was 16 h: 8 h. A sample of 2 mL of sample was collected every day for test of different parameter test.
2. Materials and methods 2.1. Culture collection and cultivation Active bacterial cultures of Pseudomonas aeruginosa (MCC No. 2622) and Bacillus cereus (MCC No. 2240) were procured from National Center for Cell Science, Pune, India. Both the bacterial cultures were cultivated in nutrient agar with sodium chloride media for 48 h at 25 ± 1 °C in incubator. After incubation period of 48 h, cultures were transferred in a single media to develop an efficient bacterial consortium. Scenedesmus abundans (NCIM No. 2897) was obtained from National chemical Laboratory, Pune, India and cultivated in BBM medium with the following components per liter: NaNO3, 0.25 g; MgSO4. 7H2O, 0.075 g; NaCl, 0.025 g; : K2HPO4, 0.075 g; KH2PO4, 0.175 g; CaCl2 2H2O, 0.025 g; H3BO3, 0.011 g; EDTA, 0.05 g; KOH, 0.031 g; FeSO4·7H2O 0.0049 g; H2SO4, 0.001 mL and trace elements (ZnSO4·7H2O, 0.00882 g MnCl2·4H2O 0.00144 g, MoO3, 0.0071 CuSO4·5H2O, 0.00157 g Co (NO3)2·6H2O 0.00049 g). The microalga Scenedesmus abundans was cultivated in 1L Erlenmeyer flasks with 750 mL of autoclaved BBM medium at 25 ± 1 °C under continues illumination (22W, LED tube light) with light intensity of 94.6 μ mol m−2 s−1. There was no supply of CO2 externally except the availability in the atmosphere.
2.4. Electrochemical and chemical analysis Open circuit voltage was monitored and recorded at regular interval of 1hr for 21 days by digital multi-meter connected to 100 Ω external 307
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Fig. 2. Lab scale image of MFCs used in this study.
resistances (R) in series. Power was calculated by using Ohm’s formula, P = V2/R where V and R are the voltage and resistance respectively. The power was normalized to the anode surface in order to calculate power density (mW/A2). Power density was calculated based on the electrode surface area as, PD = P/A where A is the total surface area of the electrode in m2. The COD removal efficiency (%) was calculated using the following equation:
COD Removal (%) =
Ci − Cf Ci
× 100
(1)
Where Ci, Cf are the initial and final COD respectively. 2.5. Algal growth Fig. 3. Percentage removal of COD during the incubation period of 21 days. DW50 + SWW50 showing highest removal of 78.6%.
The concentration of algal biomass was collected in every day basis and was measured by UV spectrophotometer (Shimadzu Corp, Japan) at 680 nm. For estimation of dry biomass by gravimetric process, the sample was collected every day from the algal cultivation chamber, centrifuged at 6200 rpm for 5 min and vacuum dried overnight at 104 °C. Biomass productivity and specific growth rate were calculated as follows:
in cell voltage over time indicates the capability of the inoculated consortia of culture to remove COD of the distillery spent wash and generate electricity. The value of the voltage generated was decreased after a steady removal efficiency of COD was attained due to unavailability of organic nutrient after 17 days. In case of 100:0 v/v sample, lowest removal efficiency of COD (66%) was observed due to the presence of high organic load in the distillery wastewater as it was very difficult to sustain microbes in high COD environment. However a longer lag phase of more than 10 days was observed for higher dilution of DSW was observed. The efficiency of COD removal is increased (78.6%) at 50:50 (v/v) due to less complexity of organic nutrient however a little decreased of organic removed efficiency (66%) was noticed in case of 25: 75 (v/v) due to non-availability of nutrients. Mixed culture as used in the present study showed better COD removal efficiency of 78.6% as compared to single strain treatment (72.84%) dairy wastewater as reported by Venkata Mohan [26]. COD values are considered as an index for survival of living organism in wastewater [27]. Lesser activity of electrogenic consortia and decrease in mobility of charge transfer due to ohmic resistance under such higher concentration of feed might be the reason of reduced current generation. The gradual decrease in efficiency of the MFC might have happened due to overloading of the substrate.
Biomass mass productivity = (final dry biomass − initial dry biomass)/ cultivation period (2) Specific growth rate (μ) = [ln(X1) − ln(X0)]/t1 − t0
(3)
Where X1 and X0 are biomass concentrations at t1 and t0 days respectively. Doubling Time = ln (2)/μ
(4)
3. Result and discussion 3.1. Wastewater treatment 3.1.1. COD removal efficiency The COD concentration in wastewater is an important factor which limits the power generation tendency. The dual chambered ‘H’ type MFC was operated for 21 days in batch mode using four different dilutions of wastewater. The correlation between power density calculated from voltage outputs and COD concentration over the entire incubation period was recorded as shown in Fig. 4. The dilution of distillery effluent with sewage wastewater (50:50 v/v) showed highest electricity generation than the other diluted distillery sample. COD removal efficiency of four different samples varied in between 66% to 78.66%. The steady incline in COD reduction percentage with increase
3.1.2. Pollutant removal efficiency The consumption of macronutrients and micronutrients by the microorganism to enhance its growth and hence reduction of the nutrient concentration in the wastewater supports the purpose of the advanced wastewater treatment. The various physicochemical parameters as well as 308
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Fig. 4. Relation between COD removal (%) and Power density (PD) during the incubation period at different concentration A) DSW100 + SWW0, B) DSW75 + SWW25, C) DSW50 + SWW50, D) DSW25 + SWW75 of distillery spent.
complex of the metal ions inside the cell and later reduce the toxic metal ions to a non-toxic state. Among the bacterial community many research showed that bacillus genus has maximum biosorbents capacity of inorganic pollutant.
macro and micro nutrients were analyzed, before treatment of different samples as recorded in Table 1. In this study the maximum reduction percentage of TDS and TSS of the four samples DSW100 + SWW0, DSW75 + SWW25, DSW50 + SWW50, DSW25 + SWW75 were found 27, 32, 39.66, 32.66 and 64.66, 88, 97, 94, respectively as shown in Fig. 7. The TDS and TSS levels directly related to the various parameters such as EC, HRT etc. In the present study, the maximum reductions of heavy metals like Mg, P, Cd, Ca, Cu, Mn, Pb in different sample were 60.68%, 58.12%, 77.45%, 62.78%, 95.63% and 96.29%, respectively as shown in Fig. 9. Trace metals like K+, Na+, Mg2+, Mn2+, Cu2+and Ca2+, respectively play a crucial role in bacterial growth because of their major participation in the activation of various metabolic pathways like energy consumption and storage. These mechanisms of removal of heavy metals include the efflux of metal ions outside the cell, accumulation and forms
3.2. Production of electricity Four MFCs were used in this current study were fed with distillery effluent and sewage wastewater under different concentration. All the MFCs were operated with proper anaerobic condition maintained in the anodic chamber for 21 days. The output data of voltage was monitored with multi-meter and illustrated in Fig. 5. Stable voltage growth was seen after 12–14 days of incubation periods, which indicates the highest degradation of organic substances after this period. Increasing the
Table 1 Physiochemical parameters. PARAMETERS
DISTILLERY SPENT WASH (DSW)
SWW(mg/L)
DW75 + SWW25
DW50 + SWW50
DW25 + SWW75
Color Odor pH COD (mg/L) TDS (mg/L) TSS (mg/L) TS (mg/L) Total Alkalinity (mg/L) Calcium (mg/L) Total Phosphorous (mg/L) Total Potassium (mg/L)
Dark brown Unpleasant 4.5 ± 1.5 65000–75000 37500 6500 44000 2160 1850 14680 1000
Light brown Unpleasant 7 ± 0.5 870 554 410 964 450 62 5 4
Light brown Unpleasant 6.5 ± 1 (Adjusted) 59000 28300 5000 33300 1970 1420 11050 775
Light brown Unpleasant 6.5 ± 1(Adjusted) 36750 19800 3700 23500 1350 980 7350 565
Brown Unpleasant 6.5 ± 1(Adjusted) 18950 9800 1975 11780 890 510 3690 300
309
310
63.5 ± 1.5 98−62 60.3–85.7 72.84 64.8 ± 0.3 92 – 78.6 412 ± 12 – – 354.29 323.4 ± 4 130 645 (mW/m3) – 202 ± 6 50.6–67.1 151.2–283.5 124.35 123.5 ± 3 930 457 (mW/m3) 836.81 Nafion PVAA/SSA Nafion-117 Nafion-117 Nafion-117 Clayware ceramic Nafion Nafion-117 Lysinibacillussphaericus L. sphaericus SN-2 Aspergillus awamori Pseudomonas aeruginosa, Bacillus cereus Graphite Plate Carbon cloth Carbon cloth Graphite plate Graphite plate Carbon felt graphite fiber brush Graphite rod Dual Chambered Air cathode Dual chambered Single chambered Dual chambered Dual chambered Dual chambered Dual chambered
(−) denote not present.
Microorganism Wastewater Electrode
Fig. 6. Power density (PD) in different concentration of distillery spent wash. Maximum PD of 836.81 mW/m2 in DSW50 + SWW50.
Type of MFC
Table 2 Comparative study of COD removal and Electricity production with current study.
PEM
wastewater COD concentration, increased the conductivity of the solutions which might help to improve the performance of MFC [38] but extended start-up period of 8–9 days might be due to the availability of higher concentration of organic matter for metabolic reaction in distillery effluent and inappropriate growth environment for biocatalyst [28]. The output of 100% DSW also showed higher power density from than other previous studies. The peak power density and COD removal efficiency was observed as 836.81 mW/m2 and 78.66%, respectively for sample DSW50 + SW50 at optimum environment as shown in Fig. 3. The maximum voltage generation of different dilution of DSW100 + SWW0, DSW75 + SW25, DSW50 + SW50 and DSW25 + SWW75 were 346.2 mV, 414.6 mV, 745.13 mV 615.1 mV and, respectively. The mixture of DSW and SWW were utilized as an organic substrate for bacterial biocatalyst and produces electrons (e−) and protons (H+) through redox reaction which facilitating power generation Venkata Mohan et al. [26]. After 10 days of incubation period, the MFC yielded a relatively stable operating potential of 193.33 mV in 100% distillery effluent. The generation of maximum voltage and power density of 346.2 mV and 180.42 mW/m2, respectively, noticed on 17th day and the output was higher than previous studies. From Figs. 5 and 6, it can be seen that, for 50% diluted distillery effluent the voltage generation and power density were increased up to 745.13 mV and 836.81 mW/m2, respectively, on 16th day. In 25% distillery and 75% sewage treated wastewater the maximum voltage of 615.1 mV and power density of 570.23 mW/m2 on 17th day were registered. For distillery effluent diluted with SWW the output was found higher as compared to the previous studies with distillery effluents using MFCs (Table 2). This decrease of power
Distillery Distillery Alcohol Distillery Distillery Distillery post methanation distillery effluent (PMDE) Distillery
References COD Removal (%) Current density (mA/m2) Power density (mW/m2)
Fig. 5. Open circuit voltage generation of different dilution ratio distillery spent wash. Maximum OCV of 745.13 mV in DSW50 + SWW50.
[29] [30] [31] [32] [33] [28] [34] This study
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generation is due to the lower concentration of organic substances which leads the effective electron (e−) discharge. 3.3. Biomass production In general, growth of the microalgae depends upon the cultivation media, culture condition and optimized environment. In this study, the culture conditions are same but in the MFCs CO2 for algae growth was supplied from the anode chamber to the cathode chamber in which algae was cultured. It is shown in principle that gaseous CO2 generated from substrate degradation in anode chamber can be completely removed and converted into algal biomass in the cathode chamber. Furthermore, CH4 was reported not to be the main gaseous product in MFCs. Scenedesmus abundans showed the maximum biomass yield of 0.74 g/L and biomass productivity 0.038 g/Ld−1 in 50% dilution of distillery spent wash over 21 days of cultivation period as shown in Fig. 8, which indicates the maximum anaerobic digestion in anodic chamber with supply of CO2 to cathodic chamber. From Fig. 8, the almost same growth pattern was found in all the different concentrations of wastewater. Scenedesmus abundans has a lag phase of 5 days before going to the exponential phase. It attained the stationary phase between 16 and 21 days of cultivation period. Previous studies [35] showed, a promising biomass production by Scenedesmus abundans.
Fig. 8. Biomass growth (g/L) of Scenedesmus abundans in cathodic chamber for all four sample. Maximum growth of 0.74 g/Ld−1 of DSW50 + SWW50.
dioxide source with BBM medium is the most suitable for Scenedesmus abundans as used in the present work. 4. Conclusions
3.3.1. Specific growth rate and doubling time Specific growth rate and doubling time are the major parameters of algal proliferation which indicate the relationship between the adaptation and growth in a particular condition. Table 3 indicates that the specific growth rate (0.20 d−1) and doubling time (3.38) of algae Scenedesmus abundans with the combination of sample DSW50 + SWW50 is consistently higher than all the other three samples with short of doubling time. Al-shatri et al. [36] reported specific growth rate of 0.104 d−1at 25 °C for growth of Scenedesmus dimorph on in BBM medium under constant photon flux density. In the present study all the four experiments showed higher specific growth rate in comparison to the data reported by Ali Hussain for Scenedesmus abundans. Thus the combination of anaerobic treated DSW50 + SWW50 sample as carbon
In this study, the efficiency of H-type two chamber MFC using mixture of distillery wastewater and sewage wastewater as a substrate was systematically observed. The feasibility of power generation using microalgae (Scenedesmus abundans) was demonstrated. The maximum power density of (836 mW/m2) and maximum OCV of (745.13 mV) in this study were found relatively higher as compared to other reports. The production of CO2 by the degradation of organic and inorganic compounds could be used by microalgae to promote their growth as well as oxygen generation at cathode. The maximum growth of biomass 0.74 g/Ld−1 was noticed. Due to dilution of highly organic distillery wastewater with sewage wastewater and effective adaptation of both
Fig. 7. Removal (%) of TSS and TDS in four different concentration of distillery spent wash.
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Fig. 9. Microbial degradation (%) of metallic anion and micronutrients using MFC for incubation periods of 21 days. Table 3 Biomass, Specific growth rate, doubling time and Biomass Productivity of Scenedesmus abundans. Sample
Final Biomass (g L−1)
Initial Biomass (g L−1)
Specific Growth Rate (d−1)
Doubling Time
Biomass Productivity (g/L d−1)
DSW100+ SWW0 DSW75 + SW25 DSW50 + SW50 DSW75 + SWW25
0.51 0.58 0.74 0.43
0.01 0.02 0.01 0.01
0.18 0.16 0.20 0.17
3.70 4.32 3.38 3.87
0.027 0.028 0.038 0.021
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