غربالگری باکتری‌های تجزیه‌کننده نفت خام فاضلاب‌های صنعتی شیراز
محورهای موضوعی : زیست فناوری میکروبی
مهدی حسن شاهیان
1
(استاد میکروبیولوژی دانشگاه شهید باهنر کرمان، دانشکده علوم، گروه زیست شناسی)
هاجر مرادی
2
(دانشجوی ارشد میکروبیولوژی، دانشگاه شهید باهنر کرمان، دانشکده علوم، گروه زیست شناسی)
کلید واژه: نفت خام, تجزیه زیستی, هیدروکربن, جداسازی باکتری,
چکیده مقاله :
سابقه و هدف:آلودگی محیط زیست با نفت و مشتقات آن تهدیدی جدی برای سلامت انسان و محیط زیست می‌باشد. با استفاده از فرایند زیست پالایی، امکان حذف بسیاری از آلاینده‌های زیست محیطی به ویژه فاضلاب‌های صنعتی وجود دارد. این مطالعه با هدف جداسازی و تعیین ویژگی های باکتری های تجزیه کننده نفت خام در برخی ازفاضلاب های صنعتی شیراز انجام شد.مواد و روش ها: دراین پژوهش به منظور جداسازی باکتری‌های تجزیه کننده نفت خام نمونه برداری از برخی فاضلاب‌های صنعتی در شیراز انجام شد. به منظور بررسی اثر نفت خام بر روی فراوانی و تنوع جمعیت میکروبی نمونه‌های خاک، نمونه برداری از پساب و لجن نفتی انجام شد. شناسایی و شمارش باکتری‌های تجزیه کننده و هتروتروف با روش‌های رقت تدریجی و بیشترین شمارش احتمالی (MPN) انجام شد. پس از غربالگری مشخص شد که 20 سویه توانایی رشد و تجزیه نفت خام را دارند. شناسایی مقدماتی سویه‌های جداشده بر اساس آزمون های بیوشمیایی انجام شد. سپس ویژگی هایی مانند گسترش قطره، فعالیت امولسیون کنندگی و میزان رشد و تجزیه در این باکتری‌ها بررسی شد.یافته ها: بیشترین گسترش قطره، هیدروفوبیسیته سلولی، فعالیت امولسیون کنندگی در جدایه هایPE3، SN1، DS1 به ترتیب 23، 16 و 22 درصد بود. با استفاده از این نتایج هفت سویه مناسب انتخاب شدند و میزان حذف نفت خام با روش اسپکتروفتومتری و گراویمتری مشخص گردید. سپس با روش مولکولی تعیین هویت سویه سلولومیکروبیوم سلولانس(Cellulosimicrobium cellulans) دارای بیشترین میزان تجزیه نفت خام انجام شد. این سویه بهترین عملکرد در تجزیه(67 درصد) را در غلظت 5/1 درصد نشان داد. با بهینه سازی برخی عوامل موثر بر روی تجزیه نفت خام مانند کشت مخلوط، منبع کربن، منبع ازت و زمان، حداکثر تجزیه نفت خام (100 درصد) حاصل گردید.نتیجه گیری: نتایج نشان داد که با بهینه سازی شرایط میکروارگانیسم های جداشده در این پژوهش، امکان حذف و کنترل آلوگی نفتی وجود دارد.
Background & Objectives: Pollution of the environment with petroleum and its derivatives is a serious threat to human health and the environment, so treatment of these substances is of great importance. The bioremediation process is defined as the use of microorganisms to remove many industrial effluents, including petroleum products. The aim of this research is the isolation and characterization of crude oil degrading bacteria from some industrial sewage in Shiraz. Material & Methods: This study was performed to isolate crude oil degrading bacteria from some industrial areas of Shiraz. To investigate the effect of crude oil on the frequency and variety of microbial populations of soil and water sampling, identification and counting of crude-oil degrading bacteria and heterotrophic bacteria were performed by CFU and MPN methods. Preliminary identification of the strains was carried out based on biochemical characteristics. Then properties such as oil-spreading,emulsifying activity, growth and degradation rate of these bacteria were investigated. Results:After screening, it was found that 20 strains were capable of growing and decomposing crude oil. The highest oil-spreading,bacterial adhesion to hydrocarbon, emulsifying activity were related to PE3, SN1, DS1, 23%, 16% and 22%, respectively. Using these results seven suitable strains were selected and crude oil removal rates determined by spectrophotometric and gravimetric methods for each strain. One strain with the highest rate of crude oil degradation was Cellulosimicrobium cellulans by molecular method. This strain had the best efficiency of degradation (67 %) in 1.5 parentage of crude oil. By optimization of factors affecting crude oil degradation such as carbon source, nitrogen source, time and mixed culture the best degradation (100%) was take place. Conclusion: The results of this study showed that the use of these microorganisms by creating optimal conditions can be an important step in the removal and control of oil pollution.
bacterial consortium comprising bioemulsifier producer and petroleum hydrocarbon degraders.
Bioresour Technol ep. 2019; 282: 456-463.
2. Cameotra SS, Singh P. Bioremediation of oil sludge using crude biosurfactants. Int Biodeterior
Biodegradation. 2008; 62(3): 274-280.
3. Hassanshahian M, Tebyanian H, Cappello S. Isolation and characterization of two crude-oil
degrading yeast strains, arrowia lipolytica P -20 and P -32 from Persian ulf. Mar Pollut
Bull.2012; 64: 1386–1391.
4. Hassanshahian M, Emtiazi , Cappello S. Isolation and characterization of crude-oil-degrading
bacteria from the Persian ulf and the Caspian Sea. Mar Pollut Bull. 2012; 64: 7–12.
5. Hassanshahian M, Ahmadinejad M, Tebyanian H, Kariminik A. Isolation and characterization
of alkane degrading bacteria from petroleum reservoir waste water in Iran (Kerman and Tehran
provenances). Mar Pollut Bull. 2013; 73: 300–305.
6. Shen T, Pi , Bao M, Xu N, Li , Lu J. Biodegradation of different petroleum hydrocarbons by
free and immobilized microbial consortia. Environ Sci Process Impacts. 2015; 17(12):
2022-2033.
7. Sheari Moghadam. A. The History of petrochemical plants in Iran. Hirman Publicatio. 2013; 4:
245pp.
8. Hesham AE, Alrumman SA, Al-Amari JA. 16S rDNA Phylogenetic and APD–PC Analyses
of Petroleum Polycyclic Aromatic Hydrocarbons-Degrading Bacteria Enriched from
Oil-Polluted Soils. Arab J Sci Eng. 2016; 41(6): 2095-2106.
9. Balba MT, Al-Awadhi N, Al-Daher . Bioremediation of oil-contaminated soil:
microbiological methods for feasibility assessment and field evaluation. J Microbiol Methods.
1998; 32(2): 155-164.
10. Hassanshahian M, Emtiazi , Kermanshahi K, Cappello S. Comparison of oil degrading
microbial communities in sediments from the Persian ulf and Caspian Sea. Soil Sediment
Contam. 2010; 19(3): 277-291.
11. Wrenn BA, Venosa AD. Selective enumeration of aromatic and aliphatic hydrocarbon
degrading bacteria by a most-probable-number procedure. Can J Microbiol 1996 Mar 1; 42(3):
252-258.
12. ussel M, Li X, Qu M, Wu M, Liu L, Alam MM. Exploring the novel indigenous strains for
degrading the crude oil contaminants in soil sample. Int J Environ Sci Technol. 2018: 1-12.
13. Sun W, Dong , ao P, Fu M, Ta K, Li J. Microbial communities inhabiting oil-contaminated
soils from two major oilfields in Northern China: Implications for active petroleum-degrading
capacity. J Microbio. 2015 Jun 1; 53(6): 371-378.
14. Hassanshahian M, Zeynalipour M, Hosseinzadeh Musa F .Isolation and characterization of
crude oil degrading bacteria from the Persian ulf (Khorramshahr provenance). Mar Pollut
Bull. 2014; 82: 39–44.
15. Hassanshahian M, akimov MM, Denaro , enovese M, Cappello S. Using eal-Time PC
to assess changes in the crude oil degrading microbial community in contaminated
seawater mesocosms. Int Biodeterior Biodegradation. 2014; 93: 241–248.
16. Latha , Kalaivani . Bacterial degradation of crude oil by gravimetric analysis. Eur J Exp
Biol. 2012; 3(5): 2789-2795.
17. Hassanshahian M. Isolation and characterization of biosurfactant producing bacteria from
Persian ulf (Bushehr provenance). Mar Pollut Bull. 2014; 86: 361–366.
18. Dadrasnia A, Usman MM, Wei KS, Velappan D, Jamali H, Mohebali N, Ismail S. Native
soil bacterial isolate in Malaysia exhibit promising supplements on degrading organic
pollutants. Process Saf Environ Prot. 2016 Mar 1; 100: 264-271.
19. Bell KS, Philp JC, Aw DW, Christofi N. The genus Rhodococcus. J Appl Microbiol. 1998; 85
(2): 195-210.
20. Esedafe W, Fagade O, Umaru F, Akinwotu O. Bacterial Degradation of the Polycyclic
Aromatic Hydrocarbon (PAH) -Fraction of efinery Effluent. Int J Environ Bioremediat
Biodegrad. 2015; 3, 1: 23-27.
21. Chikere C, Ekwuabu C. Culture-dependent characterization of hydrocarbon utilizing bacteria
in selected crude oil-impacted sites in Bodo, Ogoniland, Nigeria. Afr J Environ Sci Tech.
2014 ; 8(6): 401-406.
22. Shaieb F, Elghazawani A, Issa A. Studies on crude oil degrading Bacteria isolated from
Libyan desert. Int J Curr Microbiol Appl Sci. 2015; 4(2): 920-927.
23. Nkem B, Halimoon N, usof F, Johari W, Zakaria M, eddy S. Isolation, identification and
diesel-oil biodegradation capacities of indigenous hydrocarbon-degrading strains of
Cellulosimicrobium cellulans and cinetobacter baumannii from tarball at Terengganu beach,
Malaysia. Mar Pollut Bull . 2016 ; 107(1): 261-268.
24. Sevilla A, Ceballos L, Ballestas I, ojas W, estrepo J, Verbel J. Biodegradation of
biodiesel-oil by Cellulosimicrobium sp. Isolated from Colombian Caribbean soils. Environ
Technol. 2018: 1-23.
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bacterial consortium comprising bioemulsifier producer and petroleum hydrocarbon degraders.
Bioresour Technol ep. 2019; 282: 456-463.
2. Cameotra SS, Singh P. Bioremediation of oil sludge using crude biosurfactants. Int Biodeterior
Biodegradation. 2008; 62(3): 274-280.
3. Hassanshahian M, Tebyanian H, Cappello S. Isolation and characterization of two crude-oil
degrading yeast strains, arrowia lipolytica P -20 and P -32 from Persian ulf. Mar Pollut
Bull.2012; 64: 1386–1391.
4. Hassanshahian M, Emtiazi , Cappello S. Isolation and characterization of crude-oil-degrading
bacteria from the Persian ulf and the Caspian Sea. Mar Pollut Bull. 2012; 64: 7–12.
5. Hassanshahian M, Ahmadinejad M, Tebyanian H, Kariminik A. Isolation and characterization
of alkane degrading bacteria from petroleum reservoir waste water in Iran (Kerman and Tehran
provenances). Mar Pollut Bull. 2013; 73: 300–305.
6. Shen T, Pi , Bao M, Xu N, Li , Lu J. Biodegradation of different petroleum hydrocarbons by
free and immobilized microbial consortia. Environ Sci Process Impacts. 2015; 17(12):
2022-2033.
7. Sheari Moghadam. A. The History of petrochemical plants in Iran. Hirman Publicatio. 2013; 4:
245pp.
8. Hesham AE, Alrumman SA, Al-Amari JA. 16S rDNA Phylogenetic and APD–PC Analyses
of Petroleum Polycyclic Aromatic Hydrocarbons-Degrading Bacteria Enriched from
Oil-Polluted Soils. Arab J Sci Eng. 2016; 41(6): 2095-2106.
9. Balba MT, Al-Awadhi N, Al-Daher . Bioremediation of oil-contaminated soil:
microbiological methods for feasibility assessment and field evaluation. J Microbiol Methods.
1998; 32(2): 155-164.
10. Hassanshahian M, Emtiazi , Kermanshahi K, Cappello S. Comparison of oil degrading
microbial communities in sediments from the Persian ulf and Caspian Sea. Soil Sediment
Contam. 2010; 19(3): 277-291.
11. Wrenn BA, Venosa AD. Selective enumeration of aromatic and aliphatic hydrocarbon
degrading bacteria by a most-probable-number procedure. Can J Microbiol 1996 Mar 1; 42(3):
252-258.
12. ussel M, Li X, Qu M, Wu M, Liu L, Alam MM. Exploring the novel indigenous strains for
degrading the crude oil contaminants in soil sample. Int J Environ Sci Technol. 2018: 1-12.
13. Sun W, Dong , ao P, Fu M, Ta K, Li J. Microbial communities inhabiting oil-contaminated
soils from two major oilfields in Northern China: Implications for active petroleum-degrading
capacity. J Microbio. 2015 Jun 1; 53(6): 371-378.
14. Hassanshahian M, Zeynalipour M, Hosseinzadeh Musa F .Isolation and characterization of
crude oil degrading bacteria from the Persian ulf (Khorramshahr provenance). Mar Pollut
Bull. 2014; 82: 39–44.
15. Hassanshahian M, akimov MM, Denaro , enovese M, Cappello S. Using eal-Time PC
to assess changes in the crude oil degrading microbial community in contaminated
seawater mesocosms. Int Biodeterior Biodegradation. 2014; 93: 241–248.
16. Latha , Kalaivani . Bacterial degradation of crude oil by gravimetric analysis. Eur J Exp
Biol. 2012; 3(5): 2789-2795.
17. Hassanshahian M. Isolation and characterization of biosurfactant producing bacteria from
Persian ulf (Bushehr provenance). Mar Pollut Bull. 2014; 86: 361–366.
18. Dadrasnia A, Usman MM, Wei KS, Velappan D, Jamali H, Mohebali N, Ismail S. Native
soil bacterial isolate in Malaysia exhibit promising supplements on degrading organic
pollutants. Process Saf Environ Prot. 2016 Mar 1; 100: 264-271.
19. Bell KS, Philp JC, Aw DW, Christofi N. The genus Rhodococcus. J Appl Microbiol. 1998; 85
(2): 195-210.
20. Esedafe W, Fagade O, Umaru F, Akinwotu O. Bacterial Degradation of the Polycyclic
Aromatic Hydrocarbon (PAH) -Fraction of efinery Effluent. Int J Environ Bioremediat
Biodegrad. 2015; 3, 1: 23-27.
21. Chikere C, Ekwuabu C. Culture-dependent characterization of hydrocarbon utilizing bacteria
in selected crude oil-impacted sites in Bodo, Ogoniland, Nigeria. Afr J Environ Sci Tech.
2014 ; 8(6): 401-406.
22. Shaieb F, Elghazawani A, Issa A. Studies on crude oil degrading Bacteria isolated from
Libyan desert. Int J Curr Microbiol Appl Sci. 2015; 4(2): 920-927.
23. Nkem B, Halimoon N, usof F, Johari W, Zakaria M, eddy S. Isolation, identification and
diesel-oil biodegradation capacities of indigenous hydrocarbon-degrading strains of
Cellulosimicrobium cellulans and cinetobacter baumannii from tarball at Terengganu beach,
Malaysia. Mar Pollut Bull . 2016 ; 107(1): 261-268.
24. Sevilla A, Ceballos L, Ballestas I, ojas W, estrepo J, Verbel J. Biodegradation of
biodiesel-oil by Cellulosimicrobium sp. Isolated from Colombian Caribbean soils. Environ
Technol. 2018: 1-23.