جداسازی و شناسایی باکتری های تجزیه کننده فنل از فاضلاب پالایشگاه نفت
محورهای موضوعی : آلودگی صنعتیرمضانعلی دیانتی تیلکی 1 , مرتضی قلعه نوئی 2 , معصومه اسلامی فر 3
1 - دانشیار گروه مهندسی بهداشت محیط و عضو هیأت علمی دانشکده بهداشت دانشگاه علوم پزشکی مازندران، ساری، ایران.
2 - کارشناسی ارشد مهندسی بهداشت محیط، دانشگاه علوم پزشکی مازندران، ساری، ایران
3 - دکترای میکروبیولوژی، دانشکده بهداشت، دانشگاه علوم پزشکی مازندران، ساری، ایران.
کلید واژه: فنل, باکتری, تصفیه فاضلاب, پالایشگاه,
چکیده مقاله :
زمینه و هدف: فنل و مشتقات آن برای همه موجودات زنده سمی بوده و در فاضلاب پالایشگاه نفت یافت میشوند. جداسازی و شناسایی باکتریها از فاضلاب پالایشگاه نفت به منظور شناسایی باکتریهای تجزیه کننده ترکیبات آروماتیک مهم است. هدف از این تحقیق جداسازی و شناسایی باکتریها از سیستم تصفیه فاضلاب پالایشگاه نفت تهران و تعیین میزان حذف فنل بوسیله باکتریها در محیط کشت حاوی فنل بود. مواد و روش ها: مطالعه از نوع تجربی- آزمایشگاهی بود. نمونه برداری از سیستم لجن فعال تصفیه خانه فاضلاب پالایشگاه نفت تهران در دو مرحله انجام شد. ابتدا باکتریها در محیط کشت حاوی فنل به مدت 3 هفته در انکوباتور دمای 30 درجه سانتیگراد به فنل خو داده شدند. سپس جداسازی و شناسایی باکتریها مطابق روش استاندارد انجام شد . با کشت باکتریهای ایزوله در محیطهای کشت حداقل حاوی غلظت های 5/0 تا 2/1گرم بر لیتر فنل به عنوان تنها منبع کربن، رشد باکتریها با اندازه گیری دانسیته اپتیک در طول موج 600 نانومتر بررسی شد. غلظت فنل به روش اسپکتروفوتومتری با استفاده از واکنشگر4-آمینوآنتی پیرین در طول موج 510 نانومتر مورد سنجش قرار گرفت. یافته ها: باکتریهای جداشده از تصفیه خانه فاضلاب پالایشگاه نفت شامل سویههای سودوموناس، اسینتوباکتر، اشرشیاکلی ، انتروکوکوس و انتروباکتر بودند. سویه سودوموناس قادر به حذف کامل فنل تا غلظت 9/0 گرم بر لیتر، سویههای اسینتو باکتر و اشرشیاکلی تا غلظت 7/0 گرم- بر لیتر فنل را به طور کامل حذف و سویههای انتروکوکوس و انتروباکتر تا غلظت 5/0 گرم بر لیتر فنل را به میزان صد در صد از محیط کشت حذف نمودند. بحث و نتیجه گیری: سودوموناس نسبت به دیگر باکتریهای جدا شده از فاضلاب پالایشگاه نفت بیشترین میزان تجزیه فنل را در زمان کمتر نشان داده است.
Background and Objective:Phenol and its derivatives are toxic to all living organism and are found in oil refinery wastewater. Isolation and identification of bacteria from oil refinery wastewater is important to identify aromatic compounds degrading bacteria. The aims of this study were isolation and identification of bacteria from Tehran oil refinery wastewater treatment system and determine amount of phenol degradation by these bacteria. Method: This experimental study was conducted by using two series of activated sludge samples collected from Tehran oil refinery wastewater treatment plant. Adaptation of bacteria to phenol was done by culturing in growth medium containing phenol at 30°C in incubator. After that isolation and identification of bacteria was done according to standard method. Isolated bacteria were cultured in growth medium containing different concentration (0.5- 1.2gL-1) of Phenol. Bacteria growth was assayed by measuring optical density at 600nm. Concentration of Phenol in the medium growth solution was measured by spectrophotometric method using 4- Amino antipyrine as color reagent at 510 nm. Findings: Pseudomonas, Acintobacter, E.coli, Enterococcus and Enterobacter spps were isolated from oil wastewater treatment plant. Pseudomonas spp. completely removed 0.9gL-1 of Phenol, Acintobacter and E.coli removed 0.7 gL-1, Enterococcus and Enterobacter removed 0.5 gL-1 of Phenol from growth solution. Discussion and conculation: Pseudomonas spp. isolated from oil refinery wastewater treatment plant has highest phenol removal rate in shorter contact time than other isolated bacteria.
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- Movahedyan, H., Khorsandi, H., Salehi, R.., Nikaeen, M., detection of phenol degrading bacteria and pseudomonas putida in activated sludge by polymerase chain reaction. Journal of Environmental Health Science & Engineering, vol. 6(2), 115-120.
- Aresta, M., Acquaviva, MI., Baruzzi, F., Cavallo, R.A., 2010. Isolation and characterization of polyphenols-degrading bacteria from olive-mill wastewaters polluted soil, World Journal of Microbiology and Biotechnology, Vol. 26, Issue 4, pp. 639–647.
- Lin, J., Reddy, M., Moorthi, V., Qoma, B.E., 2008. Bacterial removal of toxic phenols from an industrial effluent. African Journal of Biotechnology, Vol. 7 (13), pp. 2232-2238.
- Hasanshahian, M., Lashkari, M., Kheikhah, B., 1395. Isolation and identification of Phenol degrading bacteria from effluent factory of Golgohar Sirjan. J.Water and Wastewater, Vol.1, 49-56.
- Zhenghui, L., Wenyu, X., Dehao, L., Yang Peng, Z., Shusi, L., 2016. Biodegradation of Phenol by Bacteria Strain Acinetobacter Calcoaceticus PA Isolated from Phenolic Wastewater. Int. J. Environ. Res. Public Health, vol.13, pp. 300.
- Bergey, D.H., Holt, J.H., Bergeys Manual of determination Bacteriology.9thEd., Lippi cott Williams and Wikins ,Baltimore.1992
- APHA, American Public Health Association. Standard Methods for examination of water and wastewater. 22nd Ed., AWWA, WPCF, Washington D.C.
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- Arutchelvan, V., et al., 2005. Isolation and identification of novel high strength phenol degrading bacterial strains from phenol-formaldehyde resin manufacturing industrial wastewater. J. Hazard.Mater, Vol. 127, 238–243.
- Whiteley, A.S., Bailey, M.J., 2000. Bacterial community structure and physiological state within an industrial phenol bioremediation system. Appl. Environ. Microbiol, vol.66, 2400–2407.
- Hooived, M., Heederik, D.J.J., Kogevinas, M., Boffetta, P., Needham, L.L., Patterson, D.G., Bueno-de-Mesquita, H.B., 1998. Second follow-up of a dutch cohort occupationally exposed to phenoxy herbicides, chlorophenols, and contaminants. Am J Epidemiol, 147, 891-899.
- Environmental Protection Agency (EPA). Sampling and Analysis Procedure for Screening of Industrial Effluents for Priority Pollutants, EPA Cincinnati, OH, USA, 1977.
- Lin, S.H., Chuang, T.S., 1994. Combined treatment of phenolic wastewater by wet air oxidation and activated sludge. Toxicol. Environ. Chem., vol. 44, 243–258.
- Dianati tilaki, R.A., Karimpoor, S., 2010. Kinetic study on removal of phenol from water by organo-bentonite. Asian Journal of Chemistry, vol. 22. No.6. pp.4703-06.
- Tay, S.-L., Moy, B.-P., Maszenan, A., Tay, J.-H., 2005. Comparing activated sludge and aerobic granules as microbial inocula for phenol biodegradation. Appl. Microbiol. Biotechnol, vol. 67, 708–713.
- Barbosa, T.C.P., Luz, A.P., Bedin, M.L., Gabilan, N.H.., 1996. Effect of ceramic plant effluent on phenol degradation by Acinetobacter calcoaceticus. Int. Biodeter. Biodegr. vol. 37, 122.
- Paller, G., Hommel, R.K.., Kleber, H.-P., 1995. Phenol degradation by Acinetobacter calcoaceticus NCIB 8250. J. Basic. Microbiol. vol. 35, 325–335.
- Geng, A., Soh, A., Lim, C., Loke, L., 2006. Isolation and characterization of a phenol-degrading bacterium from an industrial activated sludge. Appl. Microbiol. Biotechnol, vol. 71, 728–735.
- Arutchelvan, V. et al., 2006. Kinetics of high strength phenol degradation using Bacillus brevis. J. Hazard. Mater, vol. 129, 216–222.
- Chowdhury, MAZ., AL Mahin, A., Fakhruddin, N.M., 2006. Degradation of phenol by Pseudomonas putida when supplied as the sole carbon source and in the presence of glucose. Bangladesh J Microbiol, Vol. 23, 29-33.
- Menke, B., Rehm, H.J., 1992. Degradation of mixture of monochlorophenols and phenols as substrates for free and immobilized cells of Alcaligens sp. A7-2. Appl Microbiol Biotechnol, Vol. 37, 665-661.
- Ahmad, S.A., Syed MA, Arif N.M., Abdul Shukor, MY., Shamaan, NA., 2011. Isolation, identification and characterization of elevated phenol degrading Acinetobacter sp. Strain AQ5NOL 1. Aust J Basic Appl Sci, vol. 5(8), 1035-1045.
- Quan, X., Shi, H., Zhang, Y., Wang, J., Qian, Y., 2004. Biodegradation of 2, 4-dichlorophenol and phenol in airlift inner-loop bioreactor immobilized with Achromobacter sp. Sep Purif Technol, vol. 34 (13), 97-103.
- Ahmed, A.M., Nakhla, G.F., Farooq, S.J., 1995. Phenol degradation by Pseudomonas aeruginosa, J.Environ. Sci. Health, vol. 30, 99–107.
- Chitra, S., Sekaran, G., Padmavathi, S., Gowri, C., 1995. Removal phenolic compounds from wastewater using mutant strain of Pseudominas pictorium. J. Gen. Appl. Microbiol, vol. 41, 229–237.
- Balasankar, T., Nagarajan, S., 2000. Biodegradtion of phenol by Bacillus brevis. Asian Jr. Microbiol. Biotech. Env. Sci, vol. 2, 155–158.
- Sadia Afrin, J., 2008.Isolation and Identification of Phenol and Monochlorophenols-Degrading Bacteria: Pseudomonas and Aeromonas Species. Bangladesh J Microbiol, Vol. 25, No. 1, pp. 41-44.
- El-Sayed, w., et al. 2003 .Isolation and identification of a novel strain of the genus Ochrobactrum with phenol-degrading activity. Journal of Bioscience and Bioengineering, Vol. 96, Issue 3, 310-312.
- Przybulewska, K ., Wieczorek, A ., Nowak, A ., Pochrzaszcz, M.,2006.The isolation of microorganisms capable of phenol degradation.. Polish Journa of Microbiology, vol. 55, No (1), 63-67.
- Kafilzadeh, F., et al. 2010. Isolation and identification of phenol degrading bacteria from Lake Parishan and their growth kinetic assay. African Journal of Biotechnology, Vol. 9(40), pp. 6721-6726.
- Movahedyan, H., Khorsandi, H., Salehi, R.., Nikaeen, M., detection of phenol degrading bacteria and pseudomonas putida in activated sludge by polymerase chain reaction. Journal of Environmental Health Science & Engineering, vol. 6(2), 115-120.
- Aresta, M., Acquaviva, MI., Baruzzi, F., Cavallo, R.A., 2010. Isolation and characterization of polyphenols-degrading bacteria from olive-mill wastewaters polluted soil, World Journal of Microbiology and Biotechnology, Vol. 26, Issue 4, pp. 639–647.
- Lin, J., Reddy, M., Moorthi, V., Qoma, B.E., 2008. Bacterial removal of toxic phenols from an industrial effluent. African Journal of Biotechnology, Vol. 7 (13), pp. 2232-2238.
- Hasanshahian, M., Lashkari, M., Kheikhah, B., 1395. Isolation and identification of Phenol degrading bacteria from effluent factory of Golgohar Sirjan. J.Water and Wastewater, Vol.1, 49-56.
- Zhenghui, L., Wenyu, X., Dehao, L., Yang Peng, Z., Shusi, L., 2016. Biodegradation of Phenol by Bacteria Strain Acinetobacter Calcoaceticus PA Isolated from Phenolic Wastewater. Int. J. Environ. Res. Public Health, vol.13, pp. 300.
- Bergey, D.H., Holt, J.H., Bergeys Manual of determination Bacteriology.9thEd., Lippi cott Williams and Wikins ,Baltimore.1992
- APHA, American Public Health Association. Standard Methods for examination of water and wastewater. 22nd Ed., AWWA, WPCF, Washington D.C.