جداسازی و شناسایی Proteus sp.مقاوم به کادمیوم و بررسی پتانسیل آن در حذف بیولوژیکی کادمیوم
محورهای موضوعی : آلودگی محیط زیست (آب و فاضلاب)
مرجان میرحسینی نیا
1
(کارشناس ارشد آلودگی و حفاظت محیط زیست دریا، دانشکده علوم و فنون دریایی، دانشگاه آزاد اسلامی واحد تهران شمال.)
مریم قانع
2
(استادیار، گروه زیست شناسی، واحد اسلامشهر، دانشگاه آزاد اسلامی، اسلامشهر، ایران *(مسئول مکاتبات).)
پریسا نجات خواه
3
(دانشیار، گروه بیولوژی دریا، دانشکده علوم و فنون دریایی، دانشگاه آزاد اسلامی واحد تهران شمال.)
کلید واژه: رودخانه سالور, کادمیوم, Proteus sp. HM_AF12, حذف زیستی,
چکیده مقاله :
زمینه و هدف: آلودگی اکوسیستم ها با فلزات سنگین مبحث مهمی در دنیای امروز است. حذف فلزات سنگین از مکان های آلوده با استفاده از میکروارگانیسم ها، در مقایسه با روش های شیمیایی مقرون به صرفه تر است. هدف از این مطالعه جداسازی و شناسایی باکتری های مقاوم به کادمیوم، تعیین کم ترین غلظت مهاری کادمیوم (MIC) و بررسی توانایی حذف کادمیوم توسط سویه ها بود. روش بررسی: جداسازی باکتری های مقاوم با روش غنی سازی در محیط کشت حاوی فلز کادمیوم صورت گرفت. MIC کادمیوم با روش رقیق سازی در آگار و میزان حذف کادمیوم با استفاده از دستگاه جذب اتمی بررسی شد. شناسایی با روش برگی ((Bergey و مطالعات فیلوژنتیک بر مبنای توالی ژنrRNA 16S انجام شد. یافته ها: در این مطالعه 40 سویة مقاوم به کادمیوم از رودخانه سالور در شهرستان اسلامشهر جدا شدند. نتایج نشان داد سویة ST1 که از رسوبات جدا شده بود، مقاومت بالایی به کادمیوم داشته و MIC کادمیوم برای این سویه mM 6 بود. این سویه توانایی حذف 2/65 % کادمیوم را از محلول mM 1 کادمیوم داشت. مطالعات مولکولی نشان داد که سویة جدا شده به جنس Proteus تعلق داشته و 99 % با Proteus mirabilis مشابهت دارد. سویه جدا شدهProteus sp. HM_AF12 نامیده شد و توالی نوکلئوتیدی آن در GenBank ثبت شد. این سویه توانایی تحمل محدوده وسیعی از pH (9-5/5) را داشته و در 5/7 % نمک نیز رشد می کرد. نتیجه گیری: نتایج نشان می دهد که Proteus sp. HM_AF12 می تواند یک جاذب زیستی کم هزینه و دوست دار محیط زیست باشد و کاربردهای مهمی در حذف کادمیوم از محیط های آلوده داشته باشد.
Background: The contamination of ecosystems with heavy metals is an important issue in current world. Removal of heavy metals from contaminated sites using microorganisms is a cheaper alternative to chemical technologies. The aim of present study was isolation and characterization of Cadmium resistant bacteria, determination of Minimum Inhibitory Concentration (MIC) and bio removal potential of the isolates. Materials and Methods: Isolation of Cadmium resistant bacteria was carried out by enrichment method by medium supplemented with Cadmium chloride. The minimum inhibitory concentration of Cd2+ was determined by the agar plate dilution method and the Cadmium removal evaluated by atomic absorption spectroscopy. Identification was carried out in accordance with Bergey's Manual of Systematic Bacteriology and phylogenetic analysis was performed based on 16S rRNA gene sequences. Results: A total of 40 Cadmium resistant strains were isolated from Salour River in Islamshahr. The results showed that bacterial strain ST1 isolated from sediments was highly resistant to Cadmium. The MIC of Cd2+ for selected isolate was 6 mM. The isolate was able to remove 65.2% of Cadmium at Cadmium concentration of 1 mM. Phylogenetic analysis showed that ST1 belongs to the genus Proteus with 99% similarity to Proteus mirabilis then designated as Proteus sp. HM_AF12. The strain had a wide pH tolerance of 5.5–9.0, and salt tolerance was up to 7.5% NaCl. Conclusion: The results implied that Proteus sp. HM_AF12 can be a low cost and environmental friendly bio sorbent that may have important application in Cd2+removal from polluted environment.
1. Torab-Mostaedi, M., Asadollahzadeh, M., Alireza Hemmati, A., Khosravi, A., 2013. Equilibrium, kinetic, and thermodynamic studies for biosorption of Cadmium and nickel on grapefruit peel. Journal of the Taiwan Institute of Chemical Engineers, Vol. 44, pp. 295–302.
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4. Pérez-Marín, A B., Ballester, A., González, F., Blázquez, M.L., Mu˜noz, J.A., Sáez,J., Zapata, V.M., 2008. Study of Cadmium, Zinc and Lead biosorption by orange wastes using the subsequent addition method. Bioresour. Technol, Vol. 99, pp. 8101–8106.
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12. Mishra, A., Malik, A., 2014. Novel fungal consortium for bioremediation of metals and dyes from mixed waste stream. Bioresour. Technol, Vol. 171, pp. 217-26.
13. Machado, M.D., Soares, E.V., Soares, H.M., 2010. Removal of heavy metals using a brewer’s yeast strain of Saccharomyces cerevisiae: Chemical speciation as a tool in the prediction and improving of treatment efficiency of real electroplating effluents Journal of Hazardous Materials, Vol. 180, pp. 347–353.
14. Arivalagan, P., Singaraj, D., Haridass, V., Kaliannan, T., 2014. Removal of Cadmium from aqueous solution by batch studies usingBacillus cereus. Ecological Engineering, Vol. 71, pp. 728–735.
15. Ghane, M., Tabandeh, F., Bandehpour, M., Ghane, M., 2013. Isolation and characterization of a heavy metal resistant Comamonas sp. from industrial effluents. Iranian Journal of Science & Technology, Vol. 37A2, pp. 173-179.
16. Bagheri Bejestani, F., Ghane, M., Mirhosseininia, M., Bagheri Bejestani, O., 2013. Isolation and phylogenetic analysis of zinc resistant Acinetobacter sp. and its potential for bioremediation. African journal of biotechnology, Vol. 12, no. 26, pp. 4123-28.
17. Mohammadzadeh Karakagh, R., Chorom, M., Motamedi, H., Kianpoor, K.Y., Oustan, S., 2012. Biosorption of Cd and Ni by inactivated bacteria isolated from agricultural soil treated with sewage sludge. Ecohydrology & Hydrobiology, Vol. 12, no. 3, pp. 191-198.
18. Yazdi, M., Behzad, N., 2009. Geochemical Contamination in Seyab River. Islamshahr, Iran. Environmental Sciences, Vol. 6, no. 4, pp. 55-64.
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20. Sambrok, J., Russell D.W., Maniatis, T., 2001. Molecular cloning a laboratory manual. New York: Cold Spring Harbor Laboratory Press.
21. Johncy, M., Hemambika, B., Hemapriya, J., Ajesh Kannan, V., 2010. Comparative assessment of heavy metal removal by immobilized and dead bacterial cells: A biosorption approach. Afr. J. Environ. Sci. Technol, vol. 4, no. 2, pp. 77-83.
22. Hou, Y., Cheng, K., Li, Z., Ma, X., Wei, Y., Zhang, L., Wang, Y., 2015. Biosorption of Cadmium and Manganese Using Free Cells of Klebsiella sp. Isolated from Waste Water PLoS One, Vol. 10, no. 10 e0140962.
23. Harrison, J.J., Ceri, H., Stremick, C.A., Turner, R.J., 2004. Biofilm susceptibility to metal toxicity, Environmental Microbiology, Vol. 6, no. 12, pp. 1220-1227.
24. Raja, C.E., Anbazhagan, K. and Selvam, G.S., 2006. Isolation and characterization of a metal-resistant Pseudomonas aeruginosa strain, World Journal of Microbiology and Technology, Vol. 22, pp. 577-585.
25. Wei, G., Fan, L., Zhu, W., Fu, Y., Yu, J., Tang, M., 2009. Isolation and characterization of the heavy metal resistant bacteria CCNWRS33-2 isolated from root nodule of Lespedeza cuneata in gold mine tailings in China. Hazardous Materials, Vol. 162, pp. 50–56.
26. Roane, T.M., Rensing C., Pepper, I.L., Maier, R.M., 2008. Microorganisms and Metal Pollutants. Environmental Microbiology, 2nd Edition. Elsevier Science, San Diego, CA.
27. Qing, H., Min-na, D., Hong-yan, Q., Xiang-ming X., Guo-qiang, Z., Min, Y., 2007. Detection, isolation, and identification of Cadmium-resistant bacteria based on PCR-DGGE. Journal of Environmental Sciences, Vol. 19, pp. 1114–1119.
28. Tortora, G.J., Funke, D.R., Case, C.L., 2005. Microbiology: An Introduction, 8th ed. San Fransisco, CA: Pearson Education Inc., publishing as Benjamin Cummings.
29. Krishna, M.P., Varghese, R., Babu, A.V., Mohamed Hatha, A.A., 2012. Bioaccumulation of Cadmium by Pseudomonas sp. isolated from metal polluted industrial region. Environmental Research Engineering and Management, Vol. 3, no. 61, pp. 58-64.
30. Parungao1, M.M., Tacata1, P.S., Tanayan, C.R.G., and Lorele C. Trinidad L.C., 2007. Biosorption of copper, Cadmium and lead by copper-resistant bacteria isolated from Mogpog River, Marinduque. Philippine Journal of Science, Vol. 136, no. 2, pp. 155-165.
31. Abd-Elnaby, H., Abou-Elela, G.M., El-Sersy, N.A., 2011. Cadmium resisting bacteria in Alexandria Eastern Harbor (Egypt) and optimization of Cadmium bioaccumulation by Vibrio harveyi. African Journal of Biotechnology, Vol.
10, no.17, pp. 3412-3423.
_||_1. Torab-Mostaedi, M., Asadollahzadeh, M., Alireza Hemmati, A., Khosravi, A., 2013. Equilibrium, kinetic, and thermodynamic studies for biosorption of Cadmium and nickel on grapefruit peel. Journal of the Taiwan Institute of Chemical Engineers, Vol. 44, pp. 295–302.
2. Ghoneim, M., El-Desoky, H., El-Moselhy, K, M., Amer, A., Abou El-Naga, E., Mohamedein, L., Al-Prol, A., 2014. Removal of Cadmium from aqueous solution using marine green algae, Ulva lactuca. Egyptian Journal of Aquatic Research, Vol. 40, pp. 235–242.
3. Jabbari Nezhad Kermani, A., Faezi Ghasemi , M., Khosravan, A., Farahmand, A., Shakibaie, M.R., 2010. Cadmium bioremediation by metal resistant mutated bacteria isolated from active sludge of industrial effluent. Iran. J. Environ. Health. Sci. Eng, Vol. 7, pp. 279-286.
4. Pérez-Marín, A B., Ballester, A., González, F., Blázquez, M.L., Mu˜noz, J.A., Sáez,J., Zapata, V.M., 2008. Study of Cadmium, Zinc and Lead biosorption by orange wastes using the subsequent addition method. Bioresour. Technol, Vol. 99, pp. 8101–8106.
5. Hossain, A., Aditya, G., 2013. Cadmium biosorption potential of shell dust of the fresh water invasive snail Physa acuta. J. Environ. Chem. Eng, Vol. 1, pp. 574–580.
6. Zeng , Xi., Tang, X., Liu, X., Jiang, P., 2009. Isolation identification and characterization of Cadmium-resistant Pseudomonas aeruginosa Strain E1. j.Cent.South Univ.Technol, Vol. 16, pp. 416-421.
7. Fu, F., Wang, Q., 2011. Removal of heavy metal ions from wastewaters: a review. J.Environ. Manage, Vol. 92, pp. 407–418.
8. Xiao, X., Luo, S., Zeng, G., Wei, W., Wan, Y., Chen, L., Guo, H., Cao, Z., Yang, L., Chen, J., Xi,Q., 2010. Biosorption of Cadmium by endophytic fungus (EF) Microsphaeropsis sp. LSE10 isolated from Cadmium hyper accumulator Solanum nigrum L. Bioresour.Technol, Vol. 101, pp. 1668–1674.
9. Rathinam, A., Maharshi, B., Janardhanan, S.K., Jonnalagadda, R.R., Nair, B.U., 2010. Biosorption of Cadmium metal ion from simulated wastewaters using Hypnea valentiae biomass: a kinetic and thermodynamic study. Bioresour. Technol, Vol. 101, pp. 1466–1470.
10. Malik, A., 2004. Metal bioremediation through growing cells. Environ. Int, Vol. 30, pp. 261–278.
11. Carpio, I.E., Machado-Santelli, G., Sakata, S.K, Ferreira Filho, S.S., Rodrigues, DF., 2014. Copper removal using a heavy-metal resistant microbial consortium in a fixed-bed reactor. Water Res. Vol. 1, no. 62, pp. 156-66.
12. Mishra, A., Malik, A., 2014. Novel fungal consortium for bioremediation of metals and dyes from mixed waste stream. Bioresour. Technol, Vol. 171, pp. 217-26.
13. Machado, M.D., Soares, E.V., Soares, H.M., 2010. Removal of heavy metals using a brewer’s yeast strain of Saccharomyces cerevisiae: Chemical speciation as a tool in the prediction and improving of treatment efficiency of real electroplating effluents Journal of Hazardous Materials, Vol. 180, pp. 347–353.
14. Arivalagan, P., Singaraj, D., Haridass, V., Kaliannan, T., 2014. Removal of Cadmium from aqueous solution by batch studies usingBacillus cereus. Ecological Engineering, Vol. 71, pp. 728–735.
15. Ghane, M., Tabandeh, F., Bandehpour, M., Ghane, M., 2013. Isolation and characterization of a heavy metal resistant Comamonas sp. from industrial effluents. Iranian Journal of Science & Technology, Vol. 37A2, pp. 173-179.
16. Bagheri Bejestani, F., Ghane, M., Mirhosseininia, M., Bagheri Bejestani, O., 2013. Isolation and phylogenetic analysis of zinc resistant Acinetobacter sp. and its potential for bioremediation. African journal of biotechnology, Vol. 12, no. 26, pp. 4123-28.
17. Mohammadzadeh Karakagh, R., Chorom, M., Motamedi, H., Kianpoor, K.Y., Oustan, S., 2012. Biosorption of Cd and Ni by inactivated bacteria isolated from agricultural soil treated with sewage sludge. Ecohydrology & Hydrobiology, Vol. 12, no. 3, pp. 191-198.
18. Yazdi, M., Behzad, N., 2009. Geochemical Contamination in Seyab River. Islamshahr, Iran. Environmental Sciences, Vol. 6, no. 4, pp. 55-64.
19. Holt, G.J., Krieg, R.N., Sneath A.H.P., Staley, T., Williams, T.S., 1993. Bergey’s Manual of Systematic Bacteriology. 9nd Edition. London: Williams and Willkins.
20. Sambrok, J., Russell D.W., Maniatis, T., 2001. Molecular cloning a laboratory manual. New York: Cold Spring Harbor Laboratory Press.
21. Johncy, M., Hemambika, B., Hemapriya, J., Ajesh Kannan, V., 2010. Comparative assessment of heavy metal removal by immobilized and dead bacterial cells: A biosorption approach. Afr. J. Environ. Sci. Technol, vol. 4, no. 2, pp. 77-83.
22. Hou, Y., Cheng, K., Li, Z., Ma, X., Wei, Y., Zhang, L., Wang, Y., 2015. Biosorption of Cadmium and Manganese Using Free Cells of Klebsiella sp. Isolated from Waste Water PLoS One, Vol. 10, no. 10 e0140962.
23. Harrison, J.J., Ceri, H., Stremick, C.A., Turner, R.J., 2004. Biofilm susceptibility to metal toxicity, Environmental Microbiology, Vol. 6, no. 12, pp. 1220-1227.
24. Raja, C.E., Anbazhagan, K. and Selvam, G.S., 2006. Isolation and characterization of a metal-resistant Pseudomonas aeruginosa strain, World Journal of Microbiology and Technology, Vol. 22, pp. 577-585.
25. Wei, G., Fan, L., Zhu, W., Fu, Y., Yu, J., Tang, M., 2009. Isolation and characterization of the heavy metal resistant bacteria CCNWRS33-2 isolated from root nodule of Lespedeza cuneata in gold mine tailings in China. Hazardous Materials, Vol. 162, pp. 50–56.
26. Roane, T.M., Rensing C., Pepper, I.L., Maier, R.M., 2008. Microorganisms and Metal Pollutants. Environmental Microbiology, 2nd Edition. Elsevier Science, San Diego, CA.
27. Qing, H., Min-na, D., Hong-yan, Q., Xiang-ming X., Guo-qiang, Z., Min, Y., 2007. Detection, isolation, and identification of Cadmium-resistant bacteria based on PCR-DGGE. Journal of Environmental Sciences, Vol. 19, pp. 1114–1119.
28. Tortora, G.J., Funke, D.R., Case, C.L., 2005. Microbiology: An Introduction, 8th ed. San Fransisco, CA: Pearson Education Inc., publishing as Benjamin Cummings.
29. Krishna, M.P., Varghese, R., Babu, A.V., Mohamed Hatha, A.A., 2012. Bioaccumulation of Cadmium by Pseudomonas sp. isolated from metal polluted industrial region. Environmental Research Engineering and Management, Vol. 3, no. 61, pp. 58-64.
30. Parungao1, M.M., Tacata1, P.S., Tanayan, C.R.G., and Lorele C. Trinidad L.C., 2007. Biosorption of copper, Cadmium and lead by copper-resistant bacteria isolated from Mogpog River, Marinduque. Philippine Journal of Science, Vol. 136, no. 2, pp. 155-165.
31. Abd-Elnaby, H., Abou-Elela, G.M., El-Sersy, N.A., 2011. Cadmium resisting bacteria in Alexandria Eastern Harbor (Egypt) and optimization of Cadmium bioaccumulation by Vibrio harveyi. African Journal of Biotechnology, Vol.
10, no.17, pp. 3412-3423.