Metallic wastewater treatment by sulfate reduction using anaerobic rotating biological contactor reactor under high metal loading conditions

  • Mothe Gopi Kiran
  • Kannan PakshirajanEmail author
  • Gopal Das
Research Article
Part of the following topical collections:
  1. Special Issue—Bio-based Technologies for Resource Recovery


This study was aimed at investigating the performance of anaerobic rotating biological contactor reactor treating synthetic wastewater containing a mixture of heavy metals under sulfate reducing condition. Statistically valid factorial design of experiments was carried out to understand the dynamics of metal removal using this bioreactor system. Copper removal was maximum (>98%), followed by other heavy metals at their respective low inlet concentrations. Metal loading rates less than 3.7 mg/L∙h in case of Cu(II); less than 1.69 mg/L∙h for Ni(II), Pb(II), Zn(II), Fe(III) and Cd(II) are favorable to the performance of the An-RBC reactor. Removal efficiency of the heavy metals from mixture depended on the metal species and their inlet loading concentrations. Analysis of metal precipitates formed in the sulfidogenic bioreactor by field emission scanning electron microscopy along with energy dispersive X-ray spectroscopy (FESEM-EDX) confirmed metal sulfide precipitation by SRB. All these results clearly revealed that the attached growth biofilm bioreactor is well suited for heavy metal removal from complex mixture.


Factorial design analysis Sulfate reducing bacteria Multi-metal solution Heavy metal removal Anaerobic rotating biological contactor reactor High metal loading 



Authors gratefully acknowledge Centre for the Environment, IIT Guwahati, Assam, India, for providing the facilities to carry out the reactor studies and the Central Instruments Facility, IIT Guwahati, Assam, India, for FESEM-EDX analysis of the samples. Authors also thank Mr. Partha Protim Bakal for his help with heavy metal analysis


  1. American Public Health Association (APHA) (2005). Standard Methods for the Examination of Water and Wastewater, 21st ed. American WaterWorks Association (AWWA) &Water Environment Federation (WEF), Washington D.C.Google Scholar
  2. Bai H J, Zhang Z M, Yang G E, Li B Z (2008). Bioremediation of cadmium by growing Rhodobacter sphaeroides: Kinetic characteristic and mechanism studies. Bioresource Technology, 99(16): 7716–7722CrossRefGoogle Scholar
  3. Brahmacharimayum B, Ghosh P K (2014). Sulfate bioreduction and elemental sulfur formation in a packed bed reactor. Journal of Environmental Chemical Engineering, 2(3): 1287–1293CrossRefGoogle Scholar
  4. Cao J, Li Y, Zhang G, Yang C, Cao X (2013). Effect of Fe(III) on the biotreatment of bioleaching solutions using sulfate-reducing bacteria. International Journal of Mineral Processing, 125: 27–33CrossRefGoogle Scholar
  5. Chen C Y, Lin K C, Yang D T (1997). Comparison of the relative toxicity relationships based on batch and continuous algal toxicity tests. Chemosphere, 35(9): 1959–1965CrossRefGoogle Scholar
  6. Cord-Ruwisch R (1985). A quick method for the determination of dissolved and precipitated sulfides in cultures of sulfate-reducing bacteria. Journal of Microbiological Methods, 4(1): 33–36CrossRefGoogle Scholar
  7. Dev S, Roy S, Bhattacharya J (2016). Understanding the performance of sulfate reducing bacteria based packed bed reactor by growth kinetics study and microbial profiling. Journal of Environmental Management, 177: 101–110CrossRefGoogle Scholar
  8. Dev S, Roy S, Bhattacharya J (2017). Optimization of the operation of packed bed bioreactor to improve the sulfate and metal removal from acid mine drainage. Journal of Environmental Management, 200: 135–144CrossRefGoogle Scholar
  9. Gikas P (2007). Kinetic responses of activated sludge to individual and joint nickel (Ni(II)) and cobalt (Co(II)): An isobolographic approach. Journal of Hazardous Materials, 143(1-2): 246–256CrossRefGoogle Scholar
  10. Guo J, Kang Y, Feng Y (2017). Bioassessment of heavy metal toxicity and enhancement of heavy metal removal by sulfate-reducing bacteria in the presence of zero valent iron. Journal of Environmental Management, 203(Pt 1): 278–285CrossRefGoogle Scholar
  11. Hill J W, Petrucci R H, McCreary T W, Perry S S (2005). General Chemistry. 4th Edition, Pearson Prentice Hall, Upper Saddle River.Google Scholar
  12. Hu Z, Chandran K, Grasso D, Smets B F (2004). Comparison of nitrification inhibition by metals in batch and continuous flow reactors. Water Research, 38(18): 3949–3959CrossRefGoogle Scholar
  13. Jiménez-Rodríguez A M, Durán-Barrantes M M, Borja R, Sánchez E, ColmenarejoMF, Raposo F (2009). Heavy metals removal from acid mine drainage water using biogenic hydrogen sulphide and effluent from anaerobic treatment: Effect of pH. Journal of Hazardous Materials, 165(1-3): 759–765CrossRefGoogle Scholar
  14. Jin S, Drever J I, Colberg P J (2007). Effects of copper on sulfate reduction in bacterial consortia enriched from metal-contaminated and uncontaminated sediments. Environmental Toxicology and Chemistry, 26(2): 225–230CrossRefGoogle Scholar
  15. Kaksonen A H, Puhakka J A (2007). Sulfate reduction based bioprocesses for the treatment of acid mine drainage and the recovery of metals. Engineering in Life Sciences, 7(6): 541–564CrossRefGoogle Scholar
  16. Kieu H T Q, Müller E, Horn H (2011). Heavy metal removal in anaerobic semi-continuous stirred tank reactors by a consortium of sulfate-reducing bacteria. Water Research, 45(13): 3863–3870CrossRefGoogle Scholar
  17. Kiran M G, Pakshirajan K, Das G (2016). Heavy metal removal using sulfate reducing biomass obtained from a lab scale upflow anaerobic packed bed reactor. Journal of Environmental Engineering, 142(9): C4015010CrossRefGoogle Scholar
  18. Kiran M G, Pakshirajan K, Das G (2017a). A new application of anaerobic rotating biological contactor reactor for heavy metal removal under sulfate reducing condition. Chemical Engineering Journal, 321: 67–75CrossRefGoogle Scholar
  19. Kiran M G, Pakshirajan K, Das G (2017b). Heavy metal removal from multicomponent system by sulfate reducing bacteria: Mechanism and cell surface characterization. Journal of Hazardous Materials, 324 (Part A): 62–70CrossRefGoogle Scholar
  20. Lens P N L, van den Bosch M C, Hulshoff Pol L W, Lettinga G (1998). Effect of staging on volatile fatty acid degradation in a sulfidogenic granular sludge reactor. Water Research, 32(4): 1178–1192CrossRefGoogle Scholar
  21. Madamba P S, Liboon F A (2001). Optimization of the vacuum dehydration of celery (Apium graveolens) using the response surface methodology. Drying Technology, 19(3-4): 611–626CrossRefGoogle Scholar
  22. Min X, Chai L, Zhang C, Takasaki Y, Okura T (2008). Control of metal toxicity, effluent COD and regeneration of gel beads by immobilized sulfate-reducing bacteria. Chemosphere, 72(7): 1086–1091CrossRefGoogle Scholar
  23. Montgomery D C (2004). Design and analysis of experiments, 7th ed. New York: WileyGoogle Scholar
  24. Nagpal S, Chuichulcherm S, Livingston A, Peeva L (2000). Ethanol utilization by sulfate-reducing bacteria: an experimental and modeling study. Biotechnology and Bioengineering, 70(5): 533–543CrossRefGoogle Scholar
  25. Omil F, Lens P, Hulshoff Pol L, Lettinga G (1996). Effect of upward velocity and sulphide concentration on volatile fatty acid degradation in a sulphidogenic granular sludge reactor. Process Biochemistry, 31 (7): 699–710CrossRefGoogle Scholar
  26. Pakshirajan K, Kheria S (2012). Continuous treatment of coloured industry wastewater using immobilized Phanerochaete chrysosporium in a rotating biological contactor reactor. Journal of Environmental Management, 101: 118–123CrossRefGoogle Scholar
  27. Postgate J R (1984). The Sulphate-reducing Bacteria. Cambridge: Cambridge University Press, 107–152Google Scholar
  28. Rinzema A, Lettinga G (1998). Anaerobic treatment of sulfate containing wastewater. In: Biotreatment Systems, 3: (Wise, DL, Ed). Boca Raton: CRC Press, Inc., 65–109Google Scholar
  29. Robinson-Lora M A, Brennan R A (2009). Efficient metal removal and neutralization of acid mine drainage by crab-shell chitin under batch and continuous-flow conditions. Bioresource Technology, 100(21): 5063–5071CrossRefGoogle Scholar
  30. Saifullah, Meers E, Qadir M, de Caritat P, Tack F M, Du Laing G, Zia M H (2009). EDTA-assisted Pb phytoextraction. Chemosphere, 74: 1279–1291CrossRefGoogle Scholar
  31. Sema S S, Gikas P, James G M, Brent M P, Timothy R G (2012). Comparison of single and joint effects of Zn and Cu in continuous flow and batch reactors. Journal of Chemical Technology and Biotechnology (Oxford, Oxfordshire), 87(3): 374–380CrossRefGoogle Scholar
  32. Sen M, Dastidar M G, Roy Choudhury P K (2007). Biological removal of Cr(VI) using Fusarium solani in batch and continuous modes of operation. Enzyme and Microbial Technology, 41(1-2): 51–56CrossRefGoogle Scholar
  33. Teclu D, Tivchev G, Laing M, Wallis M (2009). Determination of the elemental composition of molasses and its suitability as carbon source for growth of sulphate-reducing bacteria. Journal of Hazardous Materials, 161(2-3): 1157–1165CrossRefGoogle Scholar
  34. Utgikar V P, Chaudhary N, Koeniger A, Tabak H H, Haines J R, Govind R (2004). Toxicity of metals and metal mixtures: Analysis of concentration and time dependence for zinc and copper. Water Research, 38(17): 3651–3658CrossRefGoogle Scholar
  35. Velasco A, Ramírez M, Volke-Sepúlveda T, González-Sánchez A, Revah S (2008). Evaluation of feed COD/sulfate ratio as a control criterion for the biological hydrogen sulfide production and lead precipitation. Journal of Hazardous Materials, 151(2-3): 407–413CrossRefGoogle Scholar
  36. Villa-Gomez D K, Pakshirajan K, Maestro R, Mushi S, Lens P N L (2015). Effect of process variables on the sulfate reduction process in bioreactors treating metal-containing wastewaters: Factorial design and response surface analyses. Biodegradation, 26(4): 299–311CrossRefGoogle Scholar
  37. Wang W, LampiMA, Huang X D, Gerhardt K, Dixon D G, Greenberg B M (2009). Assessment of mixture toxicity of copper, cadmium, and phenanthrenequinone to the marine bacterium Vibrio fischeri. Environmental Toxicology, 24(2): 166–177CrossRefGoogle Scholar
  38. Widdel F (1998). Microbiology and ecology of sulfate-and sulfurreducing bacteria. In: Zehnder A, ed. Biology of Anaerobic Microorganisms. New York: WileyGoogle Scholar
  39. Zhang M, Wang H (2016). Preparation of immobilized sulfate reducing bacteria (SRB) granules for effective bioremediation of acid mine drainage and bacterial community analysis. Minerals Engineering, 92: 63–71CrossRefGoogle Scholar
  40. Zhang M, Wang H, Han X (2016). Preparation of metal-resistant immobilized sulfate reducing bacteria beads for acid mine drainage treatment. Chemosphere, 154: 215–223CrossRefGoogle Scholar

Copyright information

© Higher Education Press and Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  • Mothe Gopi Kiran
    • 1
  • Kannan Pakshirajan
    • 1
    • 2
    Email author
  • Gopal Das
    • 1
    • 3
  1. 1.Centre for the EnvironmentIndian Institute of Technology GuwahatiGuwahatiIndia
  2. 2.Department of Biosciences and BioengineeringIndian Institute of Technology GuwahatiGuwahatiIndia
  3. 3.Department of ChemistryIndian Institute of Technology GuwahatiGuwahatiIndia

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