Arsenic, iron and nitrate removal from groundwater by mixed bacterial culture and fate of arsenic-laden biosolids

  • A. K. Shakya
  • P. K. GhoshEmail author
Original Paper


Arsenic, iron and nitrate coexist in groundwater at a wide range of concentrations in various regions of the world. This study aims at investigating the concurrent arsenic and iron removal by combining the advantages of nitrate removal in a sulphidogenic bioreactor. A laboratory-scale suspended growth reactor was used to assess the performance of mixed bacterial culture at different arsenic, iron and nitrate concentrations. A semi-batch reactor (SmBR) was operated for more than 400 days in anoxic conditions at 30 ± 4 °C with different influent arsenate (250–1000 µg/L as arsenic), iron (2.0 mg/L) and nitrate (100–250 mg/L) concentrations in simulated groundwater and HRT of 3–6 days. Effects of different electron donors to deliver removing power on arsenic, iron and nitrate were also investigated. Nitrate was completely removed at all tested concentrations, while concentration of arsenic and iron met drinking water standards. The reactor was also charged with actual groundwater containing arsenic (up to 226 µg/L) as well as iron (up to 8.3 mg/L) and was able to remove both the contaminants below drinking water standards after addition of sufficient amount of sulphate. Toxicity characteristics leaching procedure results indicated that leachate arsenic concentrations were below the maximum United States Environmental Protection Agency guideline value for arsenic and biosolids which did not impose any environmental hazard.

Graphical abstract


Arsenic Iron Nitrate Sulphidogenic Suspended growth TCLP 



The authors are thankful to Ministry of Drinking Water and Sanitation (MDWS), India [Project No. W-11017/44/2011-WQ], for partially supporting purchase of consumables, minor equipment and scholarship to the first author for certain period of time. The authors also highly acknowledge the help from the Central Instrument Facility (CIF), Indian Institute of Technology, Guwahati, for providing man power, various instrumental facilities, etc. Mr. L. Rahman, Mr. A. Das and other staff members of PHED, Bongaigaon, Assam, are duly acknowledged for helping in the collection of actual groundwater.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest. The help in any form received from any party has been acknowledged.

Supplementary material

13762_2018_1978_MOESM1_ESM.xlsx (14 kb)
Supplementary material 1 (XLSX 13 kb)
13762_2018_1978_MOESM2_ESM.docx (23 kb)
Supplementary material 2 (DOCX 22 kb)
13762_2018_1978_MOESM3_ESM.docx (171 kb)
Supplementary material 3 (DOCX 170 kb)


  1. Abass OK, Teng Ma, Kong S, Wang Z, Mpinda MT (2016) A novel MD-ZVI integrated approach for high arsenic groundwater decontamination and effluent immobilization. Process Saf Environ 102:190–203CrossRefGoogle Scholar
  2. Akunna JC, Bizeau C, Moletta R (1993) Nitrate and nitrite reductions with anaerobic sludge using various carbon sources: glucose, glycerol, acetic acid, lactic acid and methanol. Water Res 27:1303–1312CrossRefGoogle Scholar
  3. Altun M, Sahinkaya E, Durukan I, Bektas S, Komnitsas K (2014) Arsenic removal in a sulfidogenic fixed-bed column bioreactor. J Hazard Mater 269:31–37CrossRefGoogle Scholar
  4. Amburgey JE, Amirtharajah A (2005) Strategic filter backwashing techniques and resulting particle passage. J Environ Eng 131:535–547CrossRefGoogle Scholar
  5. APHA (2005) Standard methods for the examination of water and wastewater, 21st edn. American Public Health Association, Washington, DCGoogle Scholar
  6. ATSDR (2000) Toxicological profile for arsenic. US Department of Health and Human Services, Public Health Service Agency for Toxic Substances and Disease Registry, p 428Google Scholar
  7. Attinti R, Sarkar D, Barrett K, Datta R (2015) Adsorption of arsenic (V) from aqueous solutions by goethite/silica nanocomposite. Int J Environ Sci Technol 12:3905–3914CrossRefGoogle Scholar
  8. Baig JA, Kazi TG, Arain MB, Afridi HI, Kandhro GA, Sarfraz RA, Jamal MK, Shah AQ (2009) Evaluation of arsenic and other physico-chemical parameters of surface and ground water of Jamshoro, Pakistan. J Hazard Mater 166(2):662–669CrossRefGoogle Scholar
  9. Barringer JL, Reilly PA (2013) Arsenic in groundwater: a summary of sources and the biogeochemical and hydrogeologic factors affecting arsenic occurrence and mobility. In: Current perspectives in contaminant hydrology and water resources sustainability. InTechGoogle Scholar
  10. Battaglia-Brunet F, Crouzet C, Burnol A, Coulon S, Morin D, Joulian C (2012) Precipitation of arsenic sulphide from acidic water in a fixed-film bioreactor. Water Res 46:3923–3933CrossRefGoogle Scholar
  11. Behari JR, Prakash R (2006) Determination of total arsenic content in water by atomic absorption spectroscopy (AAS) using vapour generation assembly (VGA). Chemosphere 63(1):17–21CrossRefGoogle Scholar
  12. Bhatnagar A, Sillanpaa M (2011) A review of emerging adsorbents for nitrate removal from water. Chem Eng J 168:493–504CrossRefGoogle Scholar
  13. BIS:10500 (2012) Indian standard: drinking water: specification, 2nd revision. Bureau of Indian standard, New DelhiGoogle Scholar
  14. Brahmacharimayum B (2014) Studies on sulfate reduction to elemental sulfur under anaerobic/microaerobic conditions. Ph.D. Thesis, Indian Institute of Technology, GuwahatiGoogle Scholar
  15. Briones-Gallardo R, Escot-Espinoza V, Cervantes-González E (2017) Removing arsenic and hydrogen sulfide production using arsenic-tolerant sulfate-reducing bacteria. Int J Environ Sci Technol 14:609–622CrossRefGoogle Scholar
  16. Chaturvedi R, Banerjee S, Chattopadhyay P, Bhattacharjee CR, Raul P, Borah K (2014) High iron accumulation in hair and nail of people living in iron affected areas of Assam, India. Ecotoxicol Environ Safe 110:216–220CrossRefGoogle Scholar
  17. Clancy TM, Hayes KF, Raskin L (2013) Arsenic waste management: a critical review of testing and disposal of arsenic-bearing solid wastes generated during arsenic removal from drinking water. Environ Sci Technol 47:10799–10812CrossRefGoogle Scholar
  18. DeSantis TZ, Hugenholtz P, Larsen N, Rojas M, Brodie EL, Keller K, Huber T, Dalevi D, Hu P, Andersen GL (2006) Greengenes, a chimera-checked 16S rRNA gene database and workbench compatible with ARB. Appl Environ Microbiol 72:5069–5072CrossRefGoogle Scholar
  19. EA (2002) Guidance on whether wastes containing metals or metal compounds are regulated under the Hazaradous Waste Act, 2nd edn. In: Information paper, no. 5, Department of Environment and Heritage, Australia, Environment Australia (EA), pp 1–22Google Scholar
  20. Frunzo L, Esposito G, Pirozzi F, Lens P (2012) Dynamic mathematical modeling of sulfate reducing gas-lift reactors. Process Biochem 47:2172–2181CrossRefGoogle Scholar
  21. Ghanbari F, Moradi M, Mohseni-Bandpei A, Gohari F, Abkenar TM, Aghayani E (2014) Simultaneous application of iron and aluminum anodes for nitrate removal: a comprehensive parametric study. Int J Environ Sci Technol 11:1653–1660CrossRefGoogle Scholar
  22. Ghosh A (2013) Studies on microbial reduction of perchlorate in batch and continuous system. Ph.D. Thesis, Indian Institute of Technology, GuwahatiGoogle Scholar
  23. Giménez MC, Blanes PS, Buchhamer EE, Osicka RM, Morisio Y, Farías SS (2013) Assessment of heavy metals concentration in arsenic contaminated groundwater of the Chaco Plain, Argentina. Environ Chem 2013:1–12Google Scholar
  24. Hao OJ, Chen JM, Huang L, Buglass RL (1996) Sulfate-reducing bacteria. Crit Rev Environ Sci Technol 26:155–187CrossRefGoogle Scholar
  25. Henke K (2009) Arsenic: environmental chemistry, health threats and waste treatment. Wiley, Chichester. CrossRefGoogle Scholar
  26. Hooper K et al (1998) Toxicity characteristic leaching procedure fails to extract oxoanion-forming elements that are extracted by municipal solid waste leachates. Environ Sci Technol 32:3825–3830CrossRefGoogle Scholar
  27. Jadhav SV, Bringas E, Yadav GD, Rathod VK, Ortiz I, Marathe KV (2015) Arsenic and Fluoride contaminated groundwaters: a review of current technologies or contaminant removal. J Environ Manag 162:306–325CrossRefGoogle Scholar
  28. Jong T, Parry DL (2005) Evaluation of the stability of arsenic immobilized by microbial sulfate reduction using TCLP extractions and long-term leaching techniques. Chemosphere 60:254–265CrossRefGoogle Scholar
  29. Kirk MF, Roden EE, Crossey LJ, Brealey AJ, Spilde MN (2010) Experimental analysis of arsenic precipitation during microbial sulfate and iron reduction in model aquifer sediment reactors. Geochim Cosmochim Acta 74:2538–2555CrossRefGoogle Scholar
  30. Kleerebezem R, van Loosdrecht MCM (2007) Mixed culture biotechnology for bioenergy production. Curr Opin Biotechnol 18:207–212CrossRefGoogle Scholar
  31. Lee J, Ahn W-Y, Lee C-H (2001) Comparison of the filtration characteristics between attached and suspended growth microorganisms in submerged membrane bioreactor. Water Res 35:2435–2445CrossRefGoogle Scholar
  32. Li X, Zhang L, Wang G (2014) Genomic evidence reveals the extreme diversity and wide distribution of the arsenic-related genes in Burkholderiales. PLoS ONE 9:e92236CrossRefGoogle Scholar
  33. Li B, Pan X, Zhang D, Lee D-J, Al-Misned FA, Mortuza MG (2015) Anaerobic nitrate reduction with oxidation of Fe(II) by Citrobacter freundii strain PXL1: a potential candidate for simultaneous removal of As and nitrate from groundwater. Ecol Eng 77:196–201CrossRefGoogle Scholar
  34. Liamleam W, Annachhatre AP (2007) Electron donors for biological sulfate reduction. Biotechnol Adv 25:452–463CrossRefGoogle Scholar
  35. Liu X, Gao C, Zhang A, Jin P, Wang L, Feng L (2008) The nos gene cluster from gram-positive bacterium Geobacillus thermodenitrificans NG80-2 and functional characterization of the recombinant NosZ. FEMS Microbiol Lett 289:46–52CrossRefGoogle Scholar
  36. Liu F, Zhang G, Liu S, Fu Z, Chen J, Ma C (2018) Bioremoval of arsenic and antimony from wastewater by a mixed culture of sulfate-reducing bacteria using lactate and ethanol as carbon sources. Int Biodeterior Biodegradation 126:152–159CrossRefGoogle Scholar
  37. Lovley DR, Chapelle FH (1995) Deep subsurface microbial processes. Rev Geophys 33(3):365–381CrossRefGoogle Scholar
  38. Mayorga P, Moyano A, Anawar HM, García-Sánchez A (2013) Temporal variation of arsenic and nitrate content in groundwater of the Duero River Basin (Spain). Phys Chem Earth, Parts A/B/C 58–60:22–27CrossRefGoogle Scholar
  39. Meng X, Korfiatis GP, Jing C, Christodoulatos C (2001) Redox transformations of arsenic and iron in water treatment sludge during aging and TCLP extraction. Environ Sci Technol 35:3476–3481CrossRefGoogle Scholar
  40. Mohseni-Bandpi A, Elliott DJ, Zazouli MA (2013) Biological nitrate removal processes from drinking water supply-a review. J Environ Health Sci Eng 11:35CrossRefGoogle Scholar
  41. Onstott T, Chan E, Polizzotto M, Lanzon J, DeFlaun M (2011) Precipitation of arsenic under sulfate reducing conditions and subsequent leaching under aerobic conditions. Appl Geochem 26:269–285CrossRefGoogle Scholar
  42. Postgate J (2013) The sulfate-reducing bacteria: contemporary perspectives. Springer, BerlinGoogle Scholar
  43. Ramamoorthy S et al (2006) Desulfosporosinus lacus sp. nov., a sulfate-reducing bacterium isolated from pristine freshwater lake sediments. Int J Syst Evol Microbiol 56:2729–2736CrossRefGoogle Scholar
  44. Sarkar A, Paul B (2016) The global menace of arsenic and its conventional remediation: a critical review. Chemosphere 158:37–49CrossRefGoogle Scholar
  45. Shafiquzzaman M, Azam MS, Nakajima J, Bari QH (2010) Arsenic leaching characteristics of the sludges from iron based removal process. Desalination 261:41–45CrossRefGoogle Scholar
  46. Shakya AK, Ghosh PK (2018a) Simultaneous removal of arsenic and nitrate in absence of iron in an attached growth bioreactor to meet drinking water standards: importance of sulphate and empty bed contact time. J Clean Prod 186:304–312CrossRefGoogle Scholar
  47. Shakya AK, Ghosh PK (2018b) Simultaneous removal of arsenic, iron and nitrate in an attached growth bioreactor to meet drinking water standards: importance of sulphate and empty bed contact time. J Clean Prod 186:1011–1020CrossRefGoogle Scholar
  48. Shakya AK, Rajput P, Ghosh PK (2018) Investigation on stability and leaching characteristics of mixtures of biogenic arsenosulphides and iron sulphides formed under reduced conditions. J Hazard Mater 353:320–328CrossRefGoogle Scholar
  49. Sima J, Cao X, Zhao L, Luo Q (2015) Toxicity characteristic leaching procedure over-or under-estimates leachability of lead in phosphate-amended contaminated soils. Chemosphere 138:744–750CrossRefGoogle Scholar
  50. Snyder KV, Webster TM, Upadhyaya G, Hayes KF, Raskin L (2016) Vinegar-amended anaerobic biosand filter for the removal of arsenic and nitrate from groundwater. J Environ Manag 171:21–28CrossRefGoogle Scholar
  51. Spain AM, Krumholz LR (2011) Nitrate-reducing bacteria at the nitrate and radionuclide contaminated Oak Ridge Integrated Field research challenge site: a review. Geomicrobiol J 28:418–429CrossRefGoogle Scholar
  52. Tabelin C, Igarashi T (2009) Mechanisms of arsenic and lead release from hydrothermally altered rock. J Hazard Mater 169:980–990CrossRefGoogle Scholar
  53. Tabelin CB, Hashimoto A, Igarashi T, Yoneda T (2014) Leaching of boron, arsenic and selenium from sedimentary rocks: II. pH dependence, speciation and mechanisms of release. Sci Total Environ 473:244–253CrossRefGoogle Scholar
  54. Teunissen K, Abrahamse A, Leijssen H, Rietveld L, Dijk HV (2008) Removal of both dissolved and particulate iron from groundwater. Drink Water Eng Sci Discuss 1:87–115CrossRefGoogle Scholar
  55. Upadhyaya G, Jackson J, Clancy TM, Hyun SP, Brown J, Hayes KF, Raskin L (2010) Simultaneous removal of nitrate and arsenic from drinking water sources utilizing a fixed-bed bioreactor system. Water Res 44:4958–4969CrossRefGoogle Scholar
  56. USEPA (1986) Hazardous waste management system; land disposal restriction. In: Appendix I to part 268: toxicity characteristics leaching procedure (TCLP), pp 40643–40654Google Scholar
  57. USEPA (1992) Test methods for evaluating solid waste, physical/chemical methods, 3rd edn. In: SW-846, Method 1311. U S Government Printing Office, Washington, DCGoogle Scholar
  58. USEPA (1996) Acid digestion of sediments, sludges and soils. Method 3050B. Accessed 23 Aug 2018
  59. USEPA (2002) Office of ground water and drinking water implementation guidance for the arsenic rule EPA Report-816-D-02-005 (I3–I4) USEPA, Cincinnati, USA, 2002Google Scholar
  60. USEPA (2004) SW-846 test method 9045D: soil and waste pH. Accessed 23 Aug 2018
  61. Venkataraman K, Uddameri V (2012) Modeling simultaneous exceedance of drinking-water standards of arsenic and nitrate in the Southern Ogallala aquifer using multinomial logistic regression. J Hydrol 458–459:16–27CrossRefGoogle Scholar
  62. Wang Z, Ma T, Zhu Y, Abass OK, Liu L, Su C, Shan H (2018) Application of siderite tailings in water-supply well for As removal: experiments and field tests. Int Biodeterior Biodegradation 128:85–93CrossRefGoogle Scholar
  63. Wrighton KC et al (2010) Bacterial community structure corresponds to performance during cathodic nitrate reduction. ISME J 4:1443–1455CrossRefGoogle Scholar

Copyright information

© Islamic Azad University (IAU) 2018

Authors and Affiliations

  1. 1.Department of Civil EngineeringIndian Institute of Technology GuwahatiGuwahatiIndia

Personalised recommendations