Document Type : Original Article


1 Department of Environmental Science, Faculty of Natural Resources and Environment, Ferdowsi University of Mashhad, Mashhad, Iran

2 Department of Biology, Faculty of Science, Ferdowsi University of Mashhad, Mashhad, Iran

3 School of Life Sciences, University of Technology Sydney, Ultimo, Australia


The soil in the Sangan iron-mining region in the east of Iran is contaminated with a high concentration of heavy metals, especially iron. The release of these pollutants into environment results in the transfer and accumulation of iron through the food chains, hence a reasonable solution is required to restore it. Bioaugmentation is an environmental friendly option to reduce the hazard effects of heavy metal in the contaminated soil. In this study, the consortia of two indigenous cyanobacteria isolated from soil of Sangan iron mining and used to bioremediate soil contaminated with iron, chromium, copper, lead, and nickel. The experiments were carried out by three treatment methods, including control soil, surface soil sprayed with cyanobacteria, and soil mixed with cyanobacteria for six months under laboratory condition. The scanning electron microscope showed the development of a network of filaments of the inoculated cyanobacteria (Oscillatoria sp. and Leptolyngbya sp.) with soil particles in both treatments. Bio-augmentation of the soil increased initial nitrogen content from 406 mg/kg in control to 664 mg/kg in soil mixed with cyanobacteria and 710 mg/kg in soil sprayed with cyanobacteria. Cyanobacteria inoculation decreased the available concentration of lead and nickel. The non-available heavy metal of soil in sample sprayed with cyanobacteria was in decreasing order: Cr > Fe > Ni > As > Pb > Cu. The maximum metal removal efficiency was 32%. In soil mixed with cyanobacteria increased in the root and hypocotyl lengths of radish and lettuce was observed compared to that in the control soil, indicated in the improvement of soil quality after bioremediation.


1. Liu, L., Li, W., Song, W. and Guo, M., 2018. Remediation techniques for heavy metal-contaminated soils: Principles and applicability. Science of the Total Environment. 633: 206-219.
2. Barkett, M.O. and Akün, E., 2018. Heavy metal contents of contaminated soils and ecological risk assessment in abandoned copper mine harbor in Yedidalga, Northern Cyprus. Environmental Earth Sciences, 77(10): 1-14.
3. He, Z.L., Yang, X.E. and Stoffella, P.J., 2005. Trace elements in agroecosystems and impacts on the environment. Journal of Trace elements in Medicine and Biology, 19(2-3): 125-140.
4. Iqbal, N., Hayat, M.T., Zeb, B.S., Abbas, Z. and Ahmed, T., 2019. Phytoremediation of Cd-Contaminated Soil and Water. In: Cadmium Toxicity and Tolerance in Plants. Academic Press, pp. 531-543.
5. Loganathan, P., Hedley, M.J. and Grace, N.D., 2008. Pasture soils contaminated with fertilizer-derived cadmium and fluorine: livestock effects. In: Reviews of Environmental Contamination and Toxicology. Springer, New York, NY., pp. 29-66.
6. Bolan, N., Kunhikrishnan, A., Thangarajan, R., Kumpiene, J., Park, J., Makino, T., Kirkham, M.B. and Scheckel, K., 2014. Remediation of heavy metal (loid) s contaminated soils–to mobilize or to immobilize?. Journal of hazardous materials, 266: 141-166.
7. Mahbub, K.R., Krishnan, K., Andrews, S., Venter, H., Naidu, R. and Megharaj, M., 2017. Bio-augmentation and nutrient amendment decrease concentration of mercury in contaminated soil. Science of The Total Environment, 576: 303-309.
8. Bhattacharya, S., Gupta, K., Debnath, S., Ghosh, U.C., Chattopadhyay, D. and Mukhopadhyay, A., 2012. Arsenic bioaccumulation in rice and edible plants and subsequent transmission through food chain in Bengal basin: a review of the perspectives for environmental health. Toxicological & Environmental Chemistry, 94(3): 429-441.
9. Beiyuan, J., Li, J.S., Tsang, D.C., Wang, L., Poon, C.S., Li, X.D. and Fendorf, S., 2017. Fate of arsenic before and after chemical-enhanced washing of an arsenic-containing soil in Hong Kong. Science of the total environment, 599: 679-688.
10. WHO, Arsenic in Drinking-water 2011. Available from:
11. Kushwaha, A., Hans, N., Kumar, S. and Rani, R., 2018. A critical review on speciation, mobilization and toxicity of lead in soil-microbe-plant system and bioremediation strategies. Ecotoxicology and environmental safety, 147: 1035-1045.
12. Zhou, Z., Huang, T., Yuan, B. and Liao, X., 2016. Remediation of nitrogen-contaminated sediment using bioreactive, thin-layer capping with biozeolite. Soil and Sediment Contamination: An International Journal, 25(1): 89-100.
13. Dassekpo, J.B.M., Ning, J. and Zha, X., 2018. Potential solidification/stabilization of clay-waste using green geopolymer remediation technologies. Process Safety and Environmental Protection, 117: 684-693.
14. Alshawabkeh, A.N., 2009. Electrokinetic soil remediation: challenges and opportunities. Separation Science and Technology, 44(10): 2171- 2187.
15. Tajudin, S.A.A., Azmi, M.A.M. and Nabila, A.T.A., 2016. Stabilization/solidification remediation method for contaminated soil: a review. In IOP Conference Series: Materials Science and Engineering (Vol. 136, No. 1), IOP Publishing. doi: 10.1088/1757- 899X/136/1/012043
16. Bertagnolli, C., Grishin, A., Vincent, T. and Guibal, E., 2016. Recovering heavy metal ions from complex solutions using polyethylenimine derivatives encapsulated in alginate matrix. Industrial & Engineering Chemistry Research, 55(8): 2461-2470.
17. Guo, X., Wei, Z., Wu, Q., Li, C., Qian, T. and Zheng, W., 2016. Effect of soil washing with only chelators or combining with ferric chloride on soil heavy metal removal and phytoavailability: field experiments. Chemosphere, 147: 412-419.
18. Sasikala, S. and Muthuraman, G., 2016. Removal of heavy metals from wastewater using Tribulus terrestris herbal plants powder. Iranian (Iranica) Journal of Energy and Environment, 7(1): 39-47.
19. Deepali, A., 2011. Bioremediation of Chromium (VI) from textile industry’s effluent and contaminated soil using Pseudomonas putida. Iranian (Iranica) Journal of Energy & Environment, 2(1): 24-31.
20. Li, X., Peng, W., Jia, Y., Lu, L. and Fan, W., 2016. Bioremediation of lead contaminated soil with Rhodobacter sphaeroides. Chemosphere, 156: 228-235.
21. Marques, A.P., Rangel, A.O. and Castro, P.M., 2011. Remediation of heavy metal contaminated soils: an overview of site remediation techniques. Critical Reviews in Environmental Science and Technology, 41(10): 879-914.
22. Wu, G., Kang, H., Zhang, X., Shao, H., Chu, L. and Ruan, C., 2010. A critical review on the bio-removal of hazardous heavy metals from contaminated soils: issues, progress, eco-environmental concerns and opportunities. Journal of Hazardous Materials, 174(1-3): 1-8.
23. Issa, O.M., Défarge, C., Le Bissonnais, Y., Marin, B., Duval, O., Bruand, A., d’Acqui, L.P., Nordenberg, S. and Annerman, M., 2007. Effects of the inoculation of cyanobacteria on the microstructure and the structural stability of a tropical soil. Plant and soil, 290(1-2): 209-219.
24. Chamizo, S., Mugnai, G., Rossi, F., Certini, G. and De Philippis, R., 2018. Cyanobacteria inoculation improves soil stability and fertility on different textured soils: gaining insights for applicability in soil restoration. Frontiers in Environmental Science, 6. https://
25. Mager, D.M. and Thomas, A.D., 2011. Extracellular polysaccharides from cyanobacterial soil crusts: a review of their role in dryland soil processes. Journal of Arid Environments, 75(2): 91-97.
26. Zhao, Y., Zhu, Q., Li, P., Zhao, L., Wang, L., Zheng, X. and Ma, H., 2014. Effects of artificially cultivated biological soil crusts on soil nutrients and biological activities in the Loess Plateau. Journal of Arid Land, 6(6): 742-752.
27. Kheirfam, H., Sadeghi, S.H. and Darki, B.Z., 2020. Soil conservation in an abandoned agricultural rain-fed land through inoculation of cyanobacteria. Catena, 187. 2019.104341
28. Roncero-Ramos, B., Román, J.R., Gómez-Serrano, C., Cantón, Y. and Acién, F.G., 2019. Production of a biocrust-cyanobacteria strain (Nostoc commune) for large-scale restoration of dryland soils. Journal of Applied Phycology, 31(4): 2217-2230.
29. Chamizo, S., Adessi, A., Certini, G. and De Philippis, R., 2020. Cyanobacteria inoculation as a potential tool for stabilization of burned soils. Restoration Ecology.
30. Kheirfam, H., 2020. Increasing soil potential for carbon sequestration using microbes from biological soil crusts. Journal of Arid Environments, 172.
31. Nisha, R., Kiran, B., Kaushik, A. and Kaushik, C.P., 2018. Bioremediation of salt affected soils using cyanobacteria in terms of physical structure, nutrient status and microbial activity. International Journal of Environmental Science and Technology, 15(3): 571-580.
32. Muñoz-Rojas, M., Román, J.R., Roncero-Ramos, B., Erickson, T.E., Merritt, D.J., Aguila-Carricondo, P. and Cantón, Y., 2018. Cyanobacteria inoculation enhances carbon sequestration in soil substrates used in dryland restoration. Science of the Total Environment, 636: 1149-1154.
33. Tiwari, S., Parihar, P., Patel, A., Singh, R. and Prasad, S.M., 2019. Metals in Cyanobacteria: Physiological and Molecular Regulation. In: Cyanobacteria. Academic Press. pp. 261-276.
34. Dabiri, R., Bakhshi Mazdeh, M. and Mollai, H., 2017. Heavy metal pollution and identification of their sources in soil over Sangan ironmining region, NE Iran. Journal of Mining and Environment, 8(2): 277- 289.
35. Kaushik, B.D., 1987. Laboratory methods for blue-green algae. Associated Publishing Company.
36. Sepehr, A., Hassanzadeh, M. and Rodriguez-Caballero, E., 2019. The protective role of cyanobacteria on soil stability in two Aridisols in northeastern Iran. Geoderma Regional, 16. 10.1016/j.geodrs.2018.e00201
37. Skinner, S. and Entwisle, T.J., 2001. Non-marine algae of Australia: 1. Survey of colonial gelatinous blue-green macroalgae (Cyanobacteria). Telopea, 9(3): 573-599.
38. Anagnostidis, K. and Komarek, J., 2005. Cyanoprokariota. Teil 2: Oscillatoriales. Band 19/2 Spektrum Akademischer Verlag Heidelberg.
39. Sant’Anna, C.L., Azevedo, M.D.P., Fiore, M.F., Lorenzi, A.S., Kaštovský, J. and Komárek, J., 2011. Subgeneric diversity of Brasilonema (cyanobacteria, Scytonemataceae). Brazilian Journal of Botany, 34(1): 51-62.
40. Wang, Q., Li, Y. and Wang, Y., 2011. Optimizing the weight loss-onignition methodology to quantify organic and carbonate carbon of sediments from diverse sources. Environmental Monitoring and Assessment, 174(1-4): 241-257.
41. Park, C.H., Li, X.R., Zhao, Y., Jia, R.L. and Hur, J.S., 2017. Rapid development of cyanobacterial crust in the field for combating desertification. PLoS One, 12(6). 10.1371/journal.pone.0179903 42. Ritchie, R.J., 2006. Consistent sets of spectrophotometric chlorophyll equations for acetone, methanol and ethanol solvents. Photosynthesis Research, 89(1): 27-41.
43. Alghanmi, S.I., Al Sulami, A.F., El-Zayat, T.A., Alhogbi, B.G. and Salam, M.A., 2015. Acid leaching of heavy metals from contaminated soil collected from Jeddah, Saudi Arabia: kinetic and thermodynamics studies. International Soil and Water Conservation Research, 3(3): 196- 208.
44. Turan, V., Khan, S.A., Iqbal, M., Ramzani, P.M.A. and Fatima, M., 2018. Promoting the productivity and quality of brinjal aligned with heavy metals immobilization in a wastewater irrigated heavy metal polluted soil with biochar and chitosan. Ecotoxicology and environmental safety, 161: 409-419.
45. Aparicio, J., Solá, M.Z.S., Benimeli, C.S., Amoroso, M.J. and Polti, M.A., 2015. Versatility of Streptomyces sp. M7 to bioremediate soils cocontaminated with Cr (VI) and lindane. Ecotoxicology and Environmental Safety, 116: 34-39.
46. Fuentes, M.S., Alvarez, A., Sáez, J.M., Benimeli, C.S. and Amoroso, M.J., 2014. Methoxychlor bioremediation by defined consortium of environmental Streptomyces strains. International Journal of Environmental Science and Technology, 11(4): 1147-1156.
47. Saez, J.M., Álvarez, A., Benimeli, C.S. and Amoroso, M.J., 2014. Enhanced lindane removal from soil slurry by immobilized Streptomyces consortium. International Biodeterioration & Biodegradation, 93: 63-69.
48. Singh, R.N., 1961. Role of blue-green algae in nitrogen economy of Indian agriculture. Indian Council of Agricultural Research, New Delhi.
49. Prabu, P.C. and Udayasoorian, C., 2007. Native cyanobacteria Westiellopsis (TL-2) sp for reclaiming paper mill effluent polluted saline sodic soil habitat of India. Electronic Journal of Environmental, Agricultural and Food Chemistry, 6(2): 1775-1786.
50. Pade, N. and Hagemann, M., 2015. Salt acclimation of cyanobacteria and their application in biotechnology. Life, 5(1): 25-49.
51. Bergi, J. and Trivedi, R., 2020. Bioremediation of Saline Soil by Cyanobacteria. In: Microbial Bioremediation & Biodegradation. Springer, Singapore. pp. 447-465.
52. Sitther, V. and Tabatabai, B., Morgan State University, 2020. Engineered cyanobacteria with enhanced salt tolerance. U.S. Patent 10,626,363.
53. Tsujimoto, R., Kamiya, N. and Fujita, Y., 2014. Transcriptional regulators ChlR and CnfR are essential for diazotrophic growth in nonheterocystous cyanobacteria. Proceedings of the National Academy of Sciences, 111(18): 6762-6767.
54. Stal, L.J. and Heyer, H., 1987. Dark anaerobic nitrogen fixation (acetylene reduction) in the cyanobacterium Oscillatoria sp. FEMS Microbiology Ecology, 3(4): 227-232.
55. Stal, L.J., Paerl, H.W., Bebout, B. and Villbrandt, M., 1994. Heterocystous versus non-heterocystous cyanobacteria in microbial mats. In Microbial Mats. Springer, Berlin, Heidelberg. pp. 403-414.
56. El-Enany, A.E. and Issa, A.A., 2000. Cyanobacteria as a biosorbent of heavy metals in sewage water. Environmental toxicology and pharmacology, 8(2): 95-101.
57. Vanhoudt, N., Vandenhove, H., Leys, N. and Janssen, P., 2018. Potential of higher plants, algae, and cyanobacteria for remediation of radioactively contaminated waters. Chemosphere, 207: 239-254.
58. De Philippis, R., Colica, G. and Micheletti, E., 2011. Exopolysaccharideproducing cyanobacteria in heavy metal removal from water: molecular basis and practical applicability of the biosorption process. Applied Microbiology and Biotechnology, 92(4): 697-708.
59. Pereira, S., Micheletti, E., Zille, A., Santos, A., Moradas-Ferreira, P., Tamagnini, P. and De Philippis, R., 2011. Using extracellular polymeric substances (EPS)-producing cyanobacteria for the bioremediation of heavy metals: do cations compete for the EPS functional groups and also accumulate inside the cell?. Microbiology, 157(2): 451-458.
60. Shukla, D., Vankar, P.S. and Srivastava, S.K., 2012. Bioremediation of hexavalent chromium by a cyanobacterial mat. Applied Water Science, 2(4): 245-251.
61. Shashirekha, V., Sridharan, M.R. and Swamy, M., 2015. Biochemical response of cyanobacterial species to trivalent chromium stress. Algal Research, 12: 421-430.
62. Faisal, M., Hameed, A. and Hasnain, S., 2005. Chromium-resistant bacteria and cyanobacteria: impact on Cr (VI) reduction potential and plant growth. Journal of Industrial Microbiology and Biotechnology, 32(11-12): 615-621.
63. Kranzler, C., Rudolf, M., Keren, N. and Schleiff, E., 2013. Iron in cyanobacteria. In Advances in botanical research (Vol. 65). Academic Press. pp. 57-105
64. Scanlan, D.J., Ostrowski, M., Mazard, S., Dufresne, A., Garczarek, L., Hess, W.R., Post, A.F., Hagemann, M., Paulsen, I. and Partensky, F., 2009. Ecological genomics of marine picocyanobacteria. Microbiology and Molecular Biology Reviews, 73(2): 249-299.
65. López-Maury, L., Sánchez-Riego, A.M., Reyes, J.C. and Florencio, F.J., 2009. The glutathione/glutaredoxin system is essential for arsenate reduction in Synechocystis sp. strain PCC 6803. Journal of Bacteriology, 191(11): 3534-3543.
66. Shaheen, R.I.F.F.A.T., Mahmud, R. and Sen, J., 2007. A study on arsenic decontaminating cyanobacteria of an arsenic affected soil. Journal of Soil and Nature, 1(2): 23-29.
67. Liu, S., Zhang, F., Chen, J. and Sun, G., 2011. Arsenic removal from contaminated soil via biovolatilization by genetically engineered bacteria under laboratory conditions. Journal of Environmental Sciences, 23(9): 1544-1550.
68. Huertas, M.J., López-Maury, L., Giner-Lamia, J., Sánchez-Riego, A.M. and Florencio, F.J., 2014. Metals in cyanobacteria: analysis of the copper, nickel, cobalt and arsenic homeostasis mechanisms. Life, 4(4): 865-886.
69. Chen, J., Dong, J., Chang, J., Guo, T., Yang, Q., Jia, W. and Shen, S., 2018. Characterization of an Hg (II)-volatilizing Pseudomonas sp. strain, DC-B1, and its potential for soil remediation when combined with biochar amendment. Ecotoxicology and environmental safety, 163: 172- 179.
70. Wang, T., Sun, H., Mao, H., Zhang, Y., Wang, C., Zhang, Z., Wang, B. and Sun, L., 2014. The immobilization of heavy metals in soil by bioaugmentation of a UV-mutant Bacillus subtilis 38 assisted by NovoGro biostimulation and changes of soil microbial community. Journal of hazardous materials, 278: 483-490.
71. Sharma, D.C., Sharma, C.P. and Tripathi, R.D., 2003. Phytotoxic lesions of chromium in maize. Chemosphere, 51(1): 63-68.
72. Polti, M.A., Atjián, M.C., Amoroso, M.J. and Abate, C.M., 2011. Soil chromium bioremediation: synergic activity of actinobacteria and plants. International Biodeterioration & Biodegradation, 65(8): 1175-1181.