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1 Bioresource Technology 124 (212) 1 7 Contents lists available at SciVerse ScienceDirect Bioresource Technology journal homepage: Investigation and optimization of the novel UASB MFC integrated system for sulfate removal and bioelectricity generation using the response surface methodology (RSM) Baogang Zhang a,, Jing Zhang a, Qi Yang a, Chuanping Feng a,, Yuling Zhu b, Zhengfang Ye c, Jinren Ni c a School of Water Resources and Environment, China University of Geosciences (Beijing), Key Laboratory of Groundwater Circulation and Evolution, Ministry of Education, Beijing 183, China b College of Life Sciences, Shaoxing University, Shaoxing 312, China c Department of Environmental Engineering, Peking University, The Key Laboratory of Water and Sediment Sciences, Ministry of Education, Beijing 1871, China highlights " COD/sulfate ratio and HRT influence the performance of UASB MFC system. " Power output and sulfate removal efficiency increase first and then decrease. " The response surface methodology is performed to optimize the system. " The simulation is undistorted and optimized results are reliable. article info abstract Article history: Received 28 May 212 Received in revised form 8 August 212 Accepted 11 August 212 Available online 19 August 212 Keywords: Microbial fuel cells Sulfate Sulfide Electricity generation Response surface methodology COD/sulfate ratio and hydraulic residence time (HRT), both of which influence sulfate loadings jointly, are recognized as the most two important affecting factors for sulfate removal and bioelectricity generation in the novel up-flow anaerobic sludge blanket reactor microbial fuel cell (UASB MFC) integrated system. The response surface methodology (RSM) was employed for the optimization of this system and the optimum condition with COD/sulfate ratio of 2.3 and HRT of 54.3 h was obtained with the target of maximizing the power output. In terms of maximizing the total sulfate removal efficiency, the obtained optimum condition was COD/sulfate ratio of 3.7 and HRT of 55.6 h. Experimental results indicated the undistorted simulation and reliable optimized results. These demonstrated that RSM was effective to evaluate and optimize the UASB MFC system for sulfate removal and energy recovery, providing a promising guide to further improvement of the system for potential applications. Ó 212 Elsevier Ltd. All rights reserved. 1. Introduction Many industrial wastewaters from sources such as sugar and paper mills (Zhang et al., 29b), contain high concentrations of biodegradable organics as well as high-strength sulfate, due to the use of sulfuric acid (Muyzer and Stams, 28). Wastewaters containing sulfate are normally treated by physicochemical and biological methods (Pant et al., 21; Sarti et al., 21). The former has some limitations including relatively high costs and energy consumption, even though may be effective (Silva et al., 22; Bayrakdar et al., 29). On the other hand, high-efficiency anaerobic technology has been employed to treat complex wastewaters such Corresponding authors. Tel.: ; fax: addresses: zbgcugb@gmail.com (B. Zhang), fengchuanping@gmail.com (C. Feng). as sulfate containing wastewater (Sarti and Zaiat, 211), due to its low operating cost, high efficiency and small footprint (Zhang et al., 211). However, hydrogen sulfide (H 2 S) with uncomfortable odor, toxicity and corrosivity is generated during the biological process, when sulfate is reduced by the sulfate-reducing bacteria (SRB) in the anaerobic bioreactors. Sulfide can lead to poisoning methanogens, hence the eventual failure of the anaerobic process and even causing problems during subsequent treatment (Parkin et al., 1991). Thus sulfide as well as H 2 S should be considered and handled before discharge. Normally, the biological oxidation of sulfide to elemental sulfur (S ) is carried out after the reduction of sulfate to sulfide in the anaerobic process (Lens et al., 1998). Aeration as well as complex operation is often needed in the biological unit. Nowadays, microbial fuel cells (MFCs), devices that use bacteria as catalysts to oxidize organic or inorganic matters and generate current, are /$ - see front matter Ó 212 Elsevier Ltd. All rights reserved.
2 2 B. Zhang et al. / Bioresource Technology 124 (212) 1 7 attracting increasing attention as they can convert chemical energy to electricity in mere one step (Logan et al., 26; He et al., 28; Zhuang et al., 21; Zhang et al., 212; Pant et al., 212). MFCs have been employed to oxidize sulfide to elemental sulfur effectively by Rabaey et al. (26) and our research group (Zhang et al., 29a). The UASB (up-flow anaerobic sludge blanket reactor) MFC coupled system has been demonstrated to treat sulfate containing wastewater with energy recovery successfully, in an easy operating form (Zhang et al., 29b). Moreover, the generated sulfide during the biological sulfate reduction can be oxidized to elemental sulfur, thus avoiding secondary pollution in the system (Zhang et al., 29b). Whereas this previous study is carried out only for the specific molasses wastewater treatment without operating parameters study, thus the affecting factors should be further investigated and the novel system should be optimized to confirm its actual applications. Response surface methodology (RSM) is a statistical method based on the multivariate non-linear model that is useful in studying interactions of various parameters affecting the process (Mundra et al., 27; Chou et al., 21). It has been widely used for optimization of affecting factors in the environmental pollution field (Halim et al., 29). COD/sulfate ratio and hydraulic residence time (HRT) are the most two important affecting factors during the anaerobic process for sulfate treatment. COD/sulfate ratio determines the amounts of organic matters and electron donors can be used by SRB to reduce sulfate to sulfide, while HRT determines the action time between pollutants and bacteria (Archilha et al., 21; Mockaitis et al., 21). Both of them influence the sulfate loading jointly and have not been investigated in previous research (Zhang et al., 29b), thus RSM can be introduced to evaluate and optimize the novel UASB MFC integrated system for sulfate removal and bioelectricity generation. In present research, the feasibility of sulfate removal and electricity generation based on the UASB MFC system was confirmed first. Subsequently, the behavior of COD/sulfate ratio and HRT as influencing factors was investigated separately. After that, RSM was performed to obtain the optimal values of the COD/sulfate ratio and HRT, aiming at maximizing power outputs and sulfate removal efficiencies, respectively. Results indicate that RSM is an effective method to evaluate and optimize the UASB MFC system for sulfate removal and energy recovery. 2. Methods 2.1. Wastewater sample and the UASB MFC system Simulated sulfate containing wastewater was applied during present research. The concentration of COD in the wastewater was fixed at 24 mg/l and was added in the form of C 6 H 12 O 6. Sulfate was added to the wastewater in the form of Na 2 SO 4 and its concentration was adjusted to form different COD/sulfate ratios. The simulated wastewater acted as the influent without additions of any other nutrient or trace metal. The schematic diagram of the proposed UASB MFC system was shown in Fig. 1. It was composed of one anaerobic unit (UASB reactor) and one MFC unit. This research was performed in the same system (omitting the aerobic unit) which had been described in detail in our previous research (Zhang et al., 29b). The UASB reactor was made of polymethyl methacrylate, with a total volume of 2.2 l (8 mm in diameter and 45 mm in height) and was inoculated with.7 l well developed anaerobic granular sludge obtained from Paques (Shanghai, China). The air cathode, single-chamber and membrane-less MFC unit built in cuboid shape was constructed using plane plexiglas, with a total volume of 1.8 l ( mm). The anode and cathode were 1 g granular graphite (1 5 mm) with a graphite rod (8 mm in diameter) and 11.5 cm 2 (exposed area) carbon paper with a catalyst layer (containing.5 mg/ cm 2 of Pt), respectively Operation of the UASB MFC system After domestication, the sulfate containing wastewater with COD of 24 mg/l and sulfate of 4 mg/l (COD/sulfate ratio of 4) was fed into the UASB MFC system, with HRT of 4 h. This lasted 3 days and the power output in the MFC unit as well as the pollutant removal efficiency was evaluated. Then the system was operated under different COD/sulfate ratios (2, 3, 5 in turn), respectively, with HRT being fixed at 4 h. Moreover, different HRTs (2, 3 and 5 h in turn) were performed, with COD/sulfate ratio of 4. After that, experiments and simulations based on RSM were carried out to investigate the operating principles and to obtain optimum values. The system was operated for at least 1 days under each condition. The MFC unit was operated at a fixed load (1 X, unless stated otherwise). Temperatures inside the two units were maintained at 3 ± 3 C by means of a water bath Analytical methods and data representation Sulfate was measured by standard barium chromate colorimetry (n = 42 nm). Sulfide was determined according to the methylene blue method (n = 665 nm) (Cline, 1969). The indication of sulfide described all species (H 2 S, HS, and S 2 ). Measurement of COD was based on digestion with potassium dichromate in concentrated sulfuric acid for 2 h at 15 C (Xu et al., 28). ph was measured by a ph-21 meter (Hanna, Italy). The open circuit voltage and voltage outputs of the MFC unit were taken at 5 min intervals throughout the whole test by linking the MFC unit to the serial communications port of a desktop personal computer via an 8-channel data acquisition system (Measurement, the USA). After the performance of the MFC unit stabilized, polarization curves were used to obtain the maximum power density by varying external resistances from 5 to 1 X using a resistor box. The MFC unit was run at least twice under each resistance to ensure the repeatability of power outputs. Current (I) was calculated at a resistance (R) from the voltage (V) by I = V/R. Power (P) was calculated by P = I V and normalized by the cathode area. Coulombic efficiency (CE) was calculated as reported previously (Lu et al., 29). 3. Results and discussion 3.1. Performance of the UASB MFC system The UASB MFC system was fed with the simulated sulfate containing wastewater with COD of 24 mg/l and sulfate of 4 mg/l (COD/sulfate ratio of 4) and operated under the HRT of 4 h for 3 days. Sulfate as well as COD could be removed, with effective energy recovery and the control of the generated sulfide during sulfate reduction. Table 1 showed that sulfate could be removed effectively in the UASB MFC system, especially in the UASB reactor, where sulfate was biologically reduced to sulfide. The sulfate removal efficiency of the UASB MFC system reached as high as 69.9 ± 1.7%, which was higher than results obtained in our previous research, where sulfate removal values were less than 6.% when actual molasses wastewater was employed as influent (Zhang et al., 29b). This could be attributed to the much easier degradation of glucose by SRB as carbon source for sulfate reduction than complex organic compounds in the molasses wastewater. Besides, MFC performed well in the aspect of hazardous by-product control in present
3 B. Zhang et al. / Bioresource Technology 124 (212) Fig. 1. Schematic diagram of the experimental system. (1) Influent tank; (2) Pump; (3) Water bath; (4) UASB reactor; (5) Water distributor; (6) Sludge blanket; (7) Baffle; (8) Gas bubble; (9) Gas liquid solid separator; (1) Biogas; (11) Water seal; (12) MFC unit; (13) Granular graphite; (14) Pt coated carbon paper; (15) Graphite rod; (16) Resistance; (17) Sample port; (18) Effluent. Table 1 Pollutants removals during the 3 d operation of the UASB MFC integrated process. The influent (mg/l) UASB reactor MFC unit The total removal efficiency (%) The effluent (mg/l) The removal efficiency (%) The effluent (mg/l) The removal efficiency (%) Sulfate ± ± ± ± 1.7 Sulfide ± ± ± ± 1.9 COD ± ± ± ± ± 2.8 study. In the MFC unit, the generated sulfide was biologically and chemically oxidized. The main oxidation product was elemental sulfur, with a small amount of sulfate, which was consistent with the previously reported research (Lee et al., 212). Sulfur element could be removed from the aqueous solution, instead of its stay in the wastewater with only valence state transformations, showing the advantage of the UASB MFC system for sulfur-bearing wastewater treatment. During the 3 days operation, the voltage outputs of the MFC unit operated in closed circuit were 4 mv, with the fixed external resistance of 1 X (Fig. 2a). The continuous current outputs were.4.6 ma correspondingly, demonstrating the successful energy recovery from sulfate containing wastewater with MFC technology. The maximum power density obtained from Fig. 2b during the operation was ± 1.5 mw/m 2, indicating the better performance of MFC in our research, compared with results from other artificial wastewater (Liu et al., 24; Jong et al., 211). The generated sulfide was beneficial to lower anode potential, since the residual sulfate and the generated sulfide acted as soluble redox mediator, which could help electrons transfer from bacterial cells to electrode surface, thus promoting the bioelectricity generation in proposed system (Ieropoulos et al., 25). Moreover, bacteria on the anode surface exhibited high electrochemical activities as reported in our previous research (Zhang et al., 29a). Preliminary results of the phylogenetic analysis indicated the main electrochemically active bacteria were Geobacter sulfurreducens and Thiobacillus, which would be further reported in our following research. In another aspect, economic analysis is essential for new technology evaluation. Common materials for MFC study were used in present system, which were also 5 higher than the cost of anaerobic digestion according to Pant et al. (211). The high cost of the electrode was the primary barrier and non-platinised cathodes were exploited to lower the electrode cost (Pant et al., 211). The capital costs of construction for the proposed system Voltage (mv) Voltage (mv) (a) Voltage outputs Time (d) (b) Current density (ma/m 2 ) 1 Fig. 2. The voltage outputs (a) and polarization curves (b) obtained from the MFC unit during the operation of the UASB MFC integrated process. could be further lowered when non-platinised cathodes were employed, which should be tested and optimized before the actual applications of the proposed system Power density (mw/m 2 )
4 4 B. Zhang et al. / Bioresource Technology 124 (212) Effect rules of COD/sulfate ratio and HRT Effect rules of the two important influencing factors for the proposed system, as COD/sulfate ratio and HRT, were evaluated, respectively. Different COD/sulfate ratios (2, 3, 4, 5, 6) were investigated first, with the fixed HRT of 4 h and COD concentration of 24 mg/l. Both maximum power density and total sulfate removal efficiency of the UASB MFC system increased first and then decreased with the gradual increase of COD/sulfate ratios as shown in Fig. 3a. The concentration of generated sulfide in the UASB reactor and in the effluent of the MFC unit might be responsible for this phenomenon. When the COD/sulfate ratio was low, the sulfate concentration in the influent was relatively high, resulting in the higher sulfide concentration in the influent of the MFC. For example, when the COD/sulfate ratio was 2, the obtained sulfide concentration in the influent of the MFC was as high as 18 ± 3.9 mg/l. Higher concentration of sulfide had toxic effect on electrochemically active bacteria (Zhao et al., 29), thus influencing the power output of the MFC unit. When the COD/sulfate ratio increased, the generated sulfide decreased, and then the potential of the anode increased while the power output was lowered accordingly. When the COD/sulfate ratio was 6, the sulfide concentration in the influent of the MFC unit was only 9 ± 1.2 mg/l. The highest maximum power density of 13.5 ± 19.2 mw/m 2 was obtained with COD/ sulfate ratio of 3. The trend of sulfate removal was similar to that of power output under different COD/sulfate ratios. When the value was low, sulfate could not be reduced fully, due to the lack of carbon source for SRB. When the COD/sulfate ratio was high, the regeneration of sulfate in the MFC unit became significant, despite of the higher reduction of sulfate in the UASB reactor. The highest total sulfate removal efficiency in the combined process was 74.1 ± 2.1%, when COD/sulfate ratio reached 3. Electricity generation and pollutant removal were also investigated under different HRTs (2, 3, 4, 5 and 6 h), with the fixed COD/sulfate ratio of 4 and the influent COD of 24 mg/l. As shown in Fig. 3b, both power output and total sulfate removal efficiency (a) Power density (mw/m 2 ) (b) Power density (mw/m 2 ) COD/Sulfate ratio Power Sulfate Power Sulfate HRT (h) Fig. 3. Power outputs and sulfate removals during the operation of the UASB MFC integrated process with different COD/sulfate ratios (a) and HRTs (b) Removal efficiency (%) Removal efficiency (%) increased first and then decreased with the increase of HRTs. When the HRT was small, there was no enough time for SRB to reduce sulfate, thus affecting the energy generation in the MFC unit and the final sulfate removal efficiency. Since the HRT was 2 h, only about 1 ± 3.2 mg/l sulfide was obtained in the UASB reactor. While the HRT was large, sulfide was in turn over-oxidized to sulfate in the MFC unit, which also influenced energy recovery and sulfate removal. When the HRT was 6 h, the total sulfate removal efficiency of 83.9 ± 1.9% was obtained in the UASB reactor. However, since sulfide, which involved in the energy generation process, was in turn oxidized to sulfate partly in the MFC unit, the final sulfate removal efficiency dropped to 72.1 ± 1.1%. Studies of influencing factors indicated that COD/sulfate ratio and HRT affected the performance of the UASB MFC system. The generation and utilization of sulfide was the key point, while the control of sulfide was also very important during the biological treatment of sulfate containing wastewater. Moreover, the generation and behavior of sulfide were related to COD/sulfate ratio and HRT, and these two factors also had close relationship. Thus it was necessary to optimize these two factors to improve the performance of the proposed system for bioelectricity generation and sulfate removal Implement of RSM for optimization RSM is an efficient method to obtain the more accurate combination of optimized conditions with less experiment, for it is able to take all the interactional factors into account, and balance the relationship between the whole and part (Teng et al., 21). RSM was designed and practical verification experiments were carried out to optimize the proposed system in present study, with maximizing voltage power densities and total sulfate removal efficiencies, respectively Target of maximizing power outputs Central composite design (CCD) (2 5 ) was carried out to obtain optimum values of COD/sulfate ratio and HRT, with the target of maximizing power outputs. The COD/sulfate ratio of 4 and HRT of 4 h were chosen as the optimum center. The experimental scheme and results were shown in Table 2. By applying multiple regression analysis on the experimental coded data, a second-order polynomial equation for power density of the MFC unit in our proposed system (Y 1 ) was obtained as follows: Y 1 ¼ 413:1 þ 413:X 1 þ 33:278X 2 42:6X 2 1 :22X2 2 3:97X 1X 2 where Y 1 represented power density (mw/m 2 ), X 1 and X 2 were coded values of COD/sulfate ratio and HRT, respectively. The polynomial equality is more accurate to describe influences of independent variables studied when the obtained R 2 is closer to 1(Liu and Tzeng, 1998). Thus, the calculated R 2 1 (.9552) obtained during present analysis implied a satisfied degree between the measured and modeled results. Moreover, results in Table 3 suggested that linear terms of X 1, X 2 and quadratic terms of X 1, X 2 had the most significant effects on power outputs (p <.5), followed by the cross product of X 1 X 2 (.5 < p <.5), while effects of other terms were insignificant (p >.5). Besides, the coefficient of X 1 was much larger than that of X 2 regardless of linear or quadratic terms in Eq. (1), indicating that COD/sulfate ratio had more influence on power outputs in the proposed process than HRT, which was consistent with findings reported before (Archilha et al., 21). Two-dimensional contour plots for the effect of the interaction variables on power outputs were generated in present study to determine the interaction among these two factors and their ð1þ
5 B. Zhang et al. / Bioresource Technology 124 (212) Table 2 The CCD for COD/sulfate ratio and HRT with the experimental responses. Run Uncoded values Coded values Experimental power density (mw/m 2 ) Experimental total sulfate removal efficiency (mw/m 2 ) COD/sulfate ratio HRT (h) COD/sulfate ratio HRT (h) ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± 1.7 Table 3 The analysis of the estimate regression coefficients for the model equation of CCD. (a) Target 1 a Target 2 b Term Coefficient p Term Coefficient p Constant Constant X X X X X 1 X X 1 X X 2 X X 2 X X 1 X X 1 X F <.1 F R R a b Maximizing power outputs. Maximizing total sulfate removal efficiencies. optimum concentration values. In this kind of plot, responses were studied by taking two variables at a time while keeping the other one at the level (Avishek and Arun, 28). Fig. 4a showed the interaction of COD/sulfate ratio and HRT when maximizing power outputs was set as the target. The elliptical contour plot showed a significant interaction between the two variables (Wang and Lu, 25). It could be seen that the response effect of HRT was less significant when compared with COD/sulfate ratio, while the interaction between them had obvious effect on power outputs. COD/ sulfate ratio might have more direct impact on sulfide generation, and sulfide could affect the behavior of the MFC as reported (Ieropoulos et al., 25). Their dominant interactions might be also attributed to the sulfide accumulation and reuse in the MFC unit. Applying point prediction feature, optimum condition with the COD/sulfate ratio of 2.3 and HRT of 54.3 h was obtained. In this condition, the predicted maximum power density of the MFC unit was mw/m 2, while the experimental result was found to be ± 22.9 mw/m 2, indicating that the simulation was undistorted and the optimized results were reliable. (b) Target of maximizing total sulfate removal efficiencies CCD (2 5 ) was also performed to maximize total sulfate removal efficiencies of the proposed system by obtaining the optimum combination of COD/sulfate ratio and HRT. The experimental scheme and results were also shown in Table 2. With the similar multiple regression analysis on the experimental coded data, a second-order polynomial equation for total sulfate removal efficiency of the proposed system (Y 2 ) was also obtained as follows: Y 2 ¼ 26:3 þ 9:1X 1 þ 1:X 2 :8X 2 1 :1X 1X 2 where Y 2 represented total sulfate removal efficiency (mw/m 2 ). ð2þ Fig. 4. Contours of mutual-influences between COD/sulfate ratio and HRT with targets of maximizing power outputs (a) and total sulfate removal efficiencies (b). The calculated R 2 2 obtained during this analysis was.933, indicating a relatively satisfied degree between the measured and modeled results. It could be seen from Table 3 that linear terms of X 1, X 2 and quadratic term of X 1 affected the total sulfate removal efficiency significantly (.5 < p <.5), while the effect of the cross product X 1 X 2 was insignificant (p >.5), indicating the less effect of the interaction between the two factors on the total sulfate removal efficiency. Unlike the Eq. (1), the coefficient of
6 6 B. Zhang et al. / Bioresource Technology 124 (212) 1 7 the quadratic term of X 2 in Eq. (2) was zero, suggesting that this quadratic term had no effect on the total sulfate removal efficiency. The coefficient of X 1 was much larger than that of X 2 regardless of linear or quadratic terms in Eq. (2), again demonstrating that COD/ sulfate ratio had more significant impact on anaerobic digestion and subsequent treatment in MFC unit for sulfate removal than HRT. Two-dimensional contour plots for the effect of these two interaction variables on total sulfate removal efficiencies were also drawn. From Fig. 4b it could be found that the elliptical shape was not obvious as Fig. 4a, which indicated that the interaction of COD/sulfate ratio and HRT was not significant when total sulfate removal efficiencies were considered. Fig. 4b also showed that COD/sulfate ratio had more significant effect on sulfate removal than HRT, just like in the aspect of bioelectricity generation. These results demonstrated again that COD/sulfate ratio had more direct relationship with sulfide which affected activities of SRB and electrochemically active bacteria (Zhao et al., 29). The obtained optimum condition was COD/sulfate ratio of 3.7 and HRT of 55.6 h, by applying point prediction feature. In this condition, the predicted maximum total sulfate removal efficiency was 71.3%, while the experimental result was found to be 72.7 ± 1.8%, indicating the reliable optimized result. It should be noted that the obtained optimum conditions were different with respective targets. Both efficiencies of energy recovery and sulfate removal should be considered when the proposed system is actually applied in the future. Moreover, COD/sulfate ratio is a fixed value for the specific sulfate containing wastewater without adjustment. Thus HRT may become the main influencing factor under this condition and the optimum condition can be determined according to the rules of single factor reported in Section 3.2 in present study. In another aspect, the proposed system was operated steadily for about 15 days during these operating factors investigation and optimization, with favorable regularities in the aspects of power outputs and total sulfate removals, demonstrating the feasibility of its long term operation in the future. Moreover, sulfide existing in the influent of the MFC unit was absorbed first on the anode surface and then was oxidized, while the residual sulfate might also been absorbed by the anode competitively, reducing the efficiency of sulfide oxidation and electricity generation. Thus most of sulfate should be biologically reduced in the UASB reactor to obtain better performance of the MFC unit. This should be further optimized combined with materials and configurations improvement before actual applications. 4. Conclusions COD/sulfate ratio and HRT were recognized as the most two important affecting factors for sulfate removal and bioelectricity generation in the UASB MFC integrated system. Both power outputs and total sulfate removal efficiencies increased first and then decreased with the increase of COD/sulfate ratio and HRT, respectively. Then RSM was performed to optimize the system, with the target of maximizing power outputs and total sulfate removal efficiencies, respectively. Results indicated that RSM was effective to evaluate and optimize the proposed system for sulfate removal and energy recovery, providing guidance to the practical application of the system in the future. Acknowledgements Financial support is gratefully acknowledged from the Fundamental Research Funds for the Central Universities (211YYL19). 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