Environmental Science and Pollution Research

, Volume 26, Issue 11, pp 10790–10799 | Cite as

Bacterial indicator reduction in dairy manure using hybrid zero-valent iron (h-ZVI) system

  • Sunghwa Han
  • Yongheng Huang
  • Zong LiuEmail author
Research Article


Novel and efficient animal wastewater treatment technologies of bacteria reduction are needed for preventing disease outbreak in animal herds and safeguarding environmental health. Zero-valent iron (ZVI) has been used to treat bacteria contaminated water for the past decades, but its passivation issue has been a major challenge. In this study, batch tests were performed to evaluate the effect of a hybrid zero-valent iron (h-ZVI) or a mixed ZVI/Fe3O4 media system on reduction of Escherichia coli (E. coli) levels. The h-ZVI media was created through a wet chemical process that uses nitrate to oxidize ZVI in the presence of externally added Fe2+ (aq.). Transforming ZVI into a h-ZVI system could overcome the passivation of ZVI and increase the reactivity of the media. The results demonstrated that E. coli cells in the bulk phase were removed rapidly by h-ZVI media. Majority of E. coli was attached (or adsorbed) to the surface of h-ZVI media within a few minutes, which suggested that adsorption was the dominant mechanism for bacterial removal in the initial phase. This adsorption was confirmed by fluorescence microscopy with CTC-DAPI double staining and transmission electron microscopy (TEM). Increasing contact time steadily inactivated E. coli; all cells were inactivated after 120 min of contact. The TEM results indicated that h-ZVI inactivated E. coli by causing direct damage on bacterial cell membrane. The results of this study strongly suggest that h-ZVI treatment can be used in water treatment industry where bacterial contamination is concerned.


Hybrid zero-valent iron (h-ZVI) Pathogen indicator reduction Manure water treatment Bacterial adsorption Bacterial inactivation Bacterial removal mechanism 



Colony forming unit


Tetrazolium salt 5-cyano-2,3-ditolyltetrazolium chloride



DDI water

Deoxygenated and deionized water

E. coli

Escherichia coli


Hybrid zero-valent iron




Optical density


Revolutions per minute


Transmission electron microscopy


Tryptic soy broth/Agar





This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors, but was supported by Texas A&M Agrilife and administered by the Department of Bio and Agricultural engineering at Texas A&M University.

Supplementary material

11356_2019_4501_MOESM1_ESM.docx (2 mb)
ESM 1 (DOCX 2069 kb)


  1. Bicudo JR, Goyal SM (2003) Pathogens and manure management systems: a review. Environ Technol 24(1):115–130Google Scholar
  2. Blowes DW, Ptacek CJ, Benner SG, McRae CWT, Bennett TA, Puls RW (2000) Treatment of inorganic contaminants using permeable reactive barriers. J Contam Hydrol 45:123–137CrossRefGoogle Scholar
  3. Btatkeu-K BD, Olvera-Vargas H, Tchatchueng JB, Noubactep C, Caré S (2014) Determining the optimum Fe0 ratio for sustainable granular Fe0/sand water filters. Chem Eng J 247:265–274CrossRefGoogle Scholar
  4. Chen J, Xiu Z, Lowry GV, Alvarez PJ (2011) Effect of natural organic matter on toxicity and reactivity of nano-scale zero-valent iron. Water Res 45(5):1995–2001CrossRefGoogle Scholar
  5. Coleman AW (1980) Enhanced detection of bacteria in natural environments by fluorochrome staining of DNA. Limnol Oceanogr 25(5):948–951CrossRefGoogle Scholar
  6. Diao M, Yao M (2009) Use of zero-valent iron nanoparticles in inactivating microbes. Water Res 43(20):5243–5251CrossRefGoogle Scholar
  7. Dos Santos Coelho F, Ardisson JD, Moura FC, Lago RM, Murad E, Fabris JD (2008) Potential application of highly reactive Fe(0)/Fe3O4 composites for the reduction of Cr(VI) environmental contaminants. Chemosphere 71(1):90–96CrossRefGoogle Scholar
  8. Edberg SC, Rice EW, Karlin RJ, Allen MJ (2000) Escherichia coli: the best biological drinking water indicator for public health protection. Journal of Applied Microbiology 88(S1):106S–116SGoogle Scholar
  9. Huang Y, Zhang TC (2005) Effects of dissolved oxygen on formation of corrosion products and concomitant oxygen and nitrate reduction in zero-valent iron systems with or without aqueous Fe2+. Water Res 39(9):1751–1760CrossRefGoogle Scholar
  10. Huang Y, Zhang TC, Shea PJ, Comfort SD (2003) Effects of oxide coating and selected cations on nitrate reduction by iron metal. J Environ Qual 32:1306–1315CrossRefGoogle Scholar
  11. Huang Y, Tang C, Zeng H (2012) Removing molybdate from water using a hybridized zero-valent iron/magnetite/Fe(II) treatment system. Chem Eng J 200-202:257–263CrossRefGoogle Scholar
  12. Huang Y, Peddi PK, Tang C, Zeng H, Teng X (2013) Hybrid zero-valent iron process for removing heavy metals and nitrate from flue-gas-desulfurization wastewater. Sep Purif Technol 118:690–698CrossRefGoogle Scholar
  13. Islam M, Doyle MP, Phatak SC, Millner P, Jiang X (2004) Persistence of enterohemorrhagic Eschierichai coli O157:H7 in soil and on leaf lettuce and parsley grown in fields treated with contaminated manure composts or irrigation water. J Food Prot 67(7):1365–1370CrossRefGoogle Scholar
  14. Keenan CR, Sedlak DL (2008) Factors affecting the yield of oxidants from the reaction of nanoparticulate zero-valent iron and oxygen. Environ Sci Technol 42:1262–1267CrossRefGoogle Scholar
  15. Keenan CR, Goth-Goldstein R, Lucas D, Sedlak DL (2009) Oxidative stress induced by zero-valent iron nanoparticles and Fe (II) in human bronchial epithelial cells. Environ Sci Technol 43:4555–4560CrossRefGoogle Scholar
  16. Kim JY, Park HJ, Lee C, Nelson KL, Sedlak DL, Yoon J (2010) Inactivation of Escherichia coli by nanoparticulate zerovalent iron and ferrous ion. Appl Environ Microbiol 76(22):7668–7670CrossRefGoogle Scholar
  17. Latour CD, Kolm HH (1976) High-gradient magnetic separation a water-treatment alternative. Water Res 68(6):325–327Google Scholar
  18. Lee C, Kim JY, Lee WI, Nelson KL, Yoon J, Sedlak DL (2008) Bactericidal effects of zero-valent iron nanoparticles on Escherichia coli. Environ Sci Technol 42:4927–4933CrossRefGoogle Scholar
  19. Liu Z, Sharara M, Gunasekaran S, Runge T (2015) Effects of large-scale manure treatment processes on pathogen reduction, protein distributions, and nutrient concentrations. Trans ASABE 59:695–702Google Scholar
  20. Liu Z, Carroll ZS, Long SC, Gunasekaran S, Runge T (2016) Use of cationic polymers to reduce pathogen levels during dairy manure separation. J Environ Manag 166:260–266CrossRefGoogle Scholar
  21. Luo J, Song G, Liu J, Qian G, Xu ZP (2014) Mechanism of enhanced nitrate reduction via micro-electrolysis at the powdered zero-valent iron/activated carbon interface. J Colloid Interface Sci 435:21–25CrossRefGoogle Scholar
  22. Macrae IC, Evans SK (1983) Factors influencing the adsorption of bacteria to magnetite in water and wastewater. Water Res 17(3):271–227CrossRefGoogle Scholar
  23. Miettinen IT, Vartiainen T, Martikainen PJ (1997) Phosphorus and bacterial growth in drinking water. Appl Environ Microbiol 63(8):3242–3245Google Scholar
  24. Morrison SJ, Metzler DR, Dwyer BP (2002) Removal of As, Mn, Se, U, V and Zn from groundwater by zero-valent iron in a passive treatment cell: reaction progress modeling. J Contam Hydrol 56:99–116CrossRefGoogle Scholar
  25. Noubactep C (2011) On the mechanism of microbe inactivation by metallic iron. J Hazard Mater 198:383–386CrossRefGoogle Scholar
  26. Noubactep C, Btatkeu KBD, Tchatchueng JB (2011) Impact of MnO2 on the efficiency of metallic iron for the removal of dissolved CrVI, CuII, MoVI, SbV, UVI and ZnII. Chem Eng J 178:78–84CrossRefGoogle Scholar
  27. Payne JB, Lawrence J (2015) Manure as a source of crop nutrients and soil amendment. Cooperative extension. Accessed 26 Oct 2015
  28. Redman AD, Macalady DL, Ahmann D (2002) Natural organic matter affects arsenic speciation and sorption onto hematite. Environ Sci Technol 36:2889–2896CrossRefGoogle Scholar
  29. Ribaudo M, Gollehon N, Aillery M, Kaplan J, Johansson R, Agapoff J, Christensen L, Breneman V, Peters M (2003) Manure management for water quality costs to animal feeding operations of applying manure nutrients to land. USDA-ERS Agricultural Economic Report 824:1–90Google Scholar
  30. Rijnaarts HHM, Norde W, Lyklema J, Zehnder AJB (1994) The isoelectric point of bacteria as an indicator for the presence of cell surface polymers that inhibit adhesion. Colloid Surface B 4:191–197CrossRefGoogle Scholar
  31. Rodriguez GG, Phipps D, Ishiguro K, Ridgway HF (1992) Use of fluorescent redox probe for direct visualization of actively respiring bacteria. Appl Environ Microbiol 58(6):1801–1808Google Scholar
  32. Scherer MM, Richter S, Valentine RL, Alvarez PJ (2000) Chemistry and microbiology of permeable reactive barriers for in situ groundwater clean up. Crit Rev Microbiol 26(4):221–264CrossRefGoogle Scholar
  33. Shi J, Ai Z, Zhang L (2014) Fe@Fe2O3 core-shell nanowires enhanced Fenton oxidation by accelerating the Fe(III)/Fe(II) cycles. Water Res 59:145–153CrossRefGoogle Scholar
  34. Shokes TE, Moller G (2000) Removal of dissolved heavy metals from acid rock drainage using iron metal. Environ Sci Technol 33:282–287CrossRefGoogle Scholar
  35. Tang C, Huang Y, Zhang Z, Chen J, Zeng H, Huang Y (2016) Rapid removal of selenate in a zero-valent iron/Fe3O4 /Fe2+ synergetic system. Appl Catal B-Environ 184:320–327CrossRefGoogle Scholar
  36. Tombácz E, Majzik A, Horvát Z, Illés E (2006) Magnetite in aqueous medium: coating its surface and surface coated with it. Rom Rep Phys 58(3):281–286Google Scholar
  37. Touati D (2000) Iron and oxidative stress in bacteria. Arch Biochem Biophys 373(1):1–6CrossRefGoogle Scholar
  38. Vanotti MB, Millner PD, Hunt PG, Ellison AQ (2005) Removal of pathogen and indicator microorganisms from liquid swine manure in multi-step biological and chemical treatment. Bioresour Technol 96(2):209–214Google Scholar
  39. Xu J, Tang J, Baig SA, Lv X, Xu X (2013) Enhanced dechlorination of 2,4-dichlorophenol by Pd/FeFe3O4 nanocomposites. J Hazard Mater 244-245:628–636CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Biological and Agricultural EngineeringTexas A&M UniversityCollege StationUSA

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