جداسازی باکتریهای تجزیه‌کننده نفت از خاک‌های آلوده نفتی پالایشگاه و پتروشیمی تبریز و شناسایی باکتریهای کارآمد

نویسندگان

1 دانشیار بیولوژی و بیوتکنولوژی خاک، گروه علوم و مهندسی خاک، دانشکده کشاورزی دانشگاه تبریز

2 دانش آموخته دکتری بیولوژی و بیوتکنولوژی خاک، گروه علوم و مهندسی خاک، دانشکده کشاورزی دانشگاه تبریز

3 استادیار شیمی کاربردی، دانشکده شیمی دانشگاه تبریز

چکیده

هیدروکربن­های نفتی از عمده­ترین آلاینده­های آلی در اکوسیستم­های خاکی در سراسر دنیا محسوب می‌شوند. استفاده از ریزجانداران برای حذف یا کاهش آلودگی نفتی، یکی از راه­های ارزان و دوستدار محیط زیست است، که جداسازی و ارزیابی کارایی باکتریها تجزیه‌کننده نفت، نخستین گام در استفاده از این روش می‌باشد. بر این اساس در این مقاله به نحوه جداسازی و ارزیابی کارایی باکتریهای نفت‌خوار پرداخته شده است. با بهره‌گیری از محیط‌های حداقل عاری از کربن (MSM) و غنی شده با نفت خام، 60 جدایه باکتری از مکان‌های آلوده پالایشگاه و پتروشیمی تبریز جداسازی شد. از این میان، کارایی 20 جدایه با استفاده از روش‌های مختلف کمی و کیفی (همانند OD و تولید بیوماس میکروبی، درصد تجزیه نفتی و تولید بیوسورفکتانت) بررسی شد. کاراترین جدایه­ها که هم دارای OD، بیومس و درصد تجزیه نفتی بالایی بودند و هم به تست‌های تولید بیوسورفکتانت پاسخ مثبت دادند شامل 10 جدایه COD2-1، COD1-5، COD4-3، COD4-2، COD9-3، COD6-3، COD7-1، COD3-1، COD1-1 و COD8-1 بودند. 10 جدایه دیگر شامل COD1-4، COD5-6، COD4-5، COD7-3، COD4-6، COD6-1، COD6-4، COD2-3، COD8-2 و COD3-3 علی‌رغم داشتن OD، بیومس و درصد تجزیه نفتی بالا توانایی تولید بیوسورفکتانت نداشتند. نتایج شناسایی مولکولی باکتری‌ها کارآمد نشان داد که این جدایه‌ها متعلق به جنس‌ها و گونه‌های Stenotrophomonas sp.،sp.  Achromobacter، Psedochrobactrum sp.، Arthrobacter sp.، Shewanella sp.، Alcaligenes sp.، sp. Pseudomonas و Acinetobacter baumannii هستند و از بین این جدایه­ها جنس‌های Stenotrophomonas sp.،Psedochrobactrum sp.، Arthrobacter sp. و Shewanella sp. با در نظر گرفتن شاخص‌های کیفی و کمی مورد مطالعه، برای آزمایشات زیست پالایی در خاکهای آلوده به نفت معرفی می‌شوند.

کلیدواژه‌ها

عنوان مقاله [English]

Isolation of Oil Degrading Bacteria from Oil Contaminated Soil Around the Oil Refinery and Petrochemical Plants of Tabriz and Identification of the Efficient Bacteria

نویسندگان [English]

  • Mohammad Reza Sarikhani 1
  • Mahnaz Afsharnia 2
  • Mahmud Zarei 3
1 Assoc. Prof. of Soil Biology and Biotechnology, Department of Soil Science, Faculty of Agriculture, University of Tabriz, Iran
2 Graduated PhD Student of Soil Biology and Biotechnology, Department of Soil Science, Faculty of Agriculture, University of Tabriz, Iran
3 Assist. Prof. of Applied Chemistry, Faculty of Chemistry, University of Tabriz, Iran
چکیده [English]

Background and Objectives
Petroleum hydrocarbons are the most common organic contaminants in water and soil ecosystems around the world. Application of soil microorganisms is one of the cost-effective and environmentally friendly approaches to remove or reduce the oil pollution. Isolation and assessment of oil-degrading bacteria is the first step in bioremediation. Accordingly, the aim of this study was isolation of crude oil degrading (COD) bacteria from contaminated soil samples around the refinery and petrochemical plants of Tabriz and assessment of their ability in oil degradation.
Methodology
In order to isolate the oil degrading bacteria from oil contaminated sludge collected from evaporation ponds of Tabriz Refinery, the carbon free minimal medium (CFMM) by enrichment methods were used. Based on bacterial growth indicators (e.g. optical density (OD) and microbial biomass, percent of crude oil degradation and production of biosurfactant), 20 efficient isolates were selected among the 60 isolates using CFMM supplemented with crude oil. All of these 20 isolates were evaluated by biosurfactant production tests. Then, 20 isolates were selected based on their growth rate, crude oil degradation potential, and their ability to degrade recalcitrant compounds (light naphta, heavy naphta, styrene and anthracene).
Findings
The most efficient strains that had the highest OD, microbial biomass and percentage of oil biodegradation with biosurfactant production ability were the isolates COD2-1, COD1-5, COD4-3, COD4-2, COD9-3, COD6-3, COD7-1, COD3-1, COD1-1 and COD8-1. While other efficient isolates (COD1-4, COD5-6, COD4-5, COD7-3, COD4-6, COD6-1, COD6-4, COD2-3, COD8-2 and COD3-3) did not produced biosurfactant. Among these bacteria, 11 efficient isolates were identified by molecular techniques. The results of molecular identification of bacteria showed that these isolates belong to the genus and species Stenotrophomonas sp., Achromobacter sp., Psedochrobactrum sp., Arthrobacter sp., Shewanella sp., Alcaligenes sp., Pseudomonas sp. and Acinetobacter baumannii.
 
Conclusion
Among the isolates, the genus Stenotrophomonas sp., Psedochrobactrum sp., Arthrobacter sp. and Shewanella sp. with high quantitative and qualitative indices in terms of bacterial growth, microbial biomass, crude oil degradation and production of biosurfactant were the best candidates for bioremediation experiments in contaminated soil.

کلیدواژه‌ها [English]

  • Biosurfactant
  • Oil bioremediation
  • Oil contaminated soil
  • Oil degrading bacteria

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دانش آب و خاک
دوره 32، شماره 4
دی 1401
صفحه 91-104

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