تعیین حساسیت به فلزات سنگین در باکتری‌های مولد سورفکتانت و کارایی آنها در حذف هیدروکربن‌های نفتی کل

نوع مقاله : مقاله پژوهشی

نویسندگان

1 گروه علوم و مهندسی خاک، دانشگاه تهران، کرج، ایران.

2 گروه مهندسی محیط زیست، دانشکده محیط زیست، دانشگاه تهران،تهران، ایران.

3 گروه علوم و مهندسی خاک، دانشکدگان کشاورزی و منابع طبیعی دانشگاه تهران، کرج، ایران

چکیده

به دلیل همراه بودن برخی از فلزات سنگین با هیدروکربن‌های نفتی، بکارگیری باکتری‌های بومی تجزیه‌کننده نفت مولد سورفکتانت که مقاوم به فلزات سنگین باشند از اولویت‌های فناوری زیست پالائی محسوب می‌شود. به همین منظور، در این مطالعه از 39 سویه تجزیه کننده آلکان‌ها و تولید کننده سورفکتانت زیستی (جداسازی شده از خاک‌های شور آلوده به نفت در مناطق نفت خیز جنوب استان خوزستان) متعلق به بانک ژن آزمایشگاه میکروبیولوژی خاک دانشگاه تهران استفاده شد. سپس حساسیت جدایه‌ها در برابر غلظت‌های مختلف (40 تا 80 ppm) فلزات سنگین (کروم، سرب، مس، نیکل، کادمیوم) مورد آزمون قرار گرفت. نتایج نشان داد فقط سه سویه Ochrobactrum lupini strain SH23، Ochrobactrum lupine strain SH24 و SH34 Bacillus subtilis subsp. Inaquosorum توانایی رشد در حضور همه فلزات سنگین را داشتند. نتایج حاصل تاثیر غلظت‌های مختلف فلزات سنگین بر MBC و MIC باکتری‌های منتخب نشان داد باکتری  Ochrobactrum lupini strain SH23 بیشترین میزان تحمل به فلزات سنگین را داشت. ارزیابی توانایی تجزیه کنندگی هیدروکربن‌های نفتی در حضور فلزات سنگین توسط باکتری‌های منتخب، نشان داد کنسرسیوم میکروبی (SH23، SH24 و SH34)  و باکتری Ochrobactrum lupini strain SH23 به ترتیب توانستند 71 و 64 درصد نفت خام با غلظت 5/0 درصد (حجمی/حجمی) را بعد از 10 روز هوادهی و در دمای 30 درجه سلسیوس تجزیه کنند. یافته‌های این پژوهش ثابت کرد با انجام مطالعات زیست‌محیطی تکمیلی می‌توان از باکتری Ochrobactrum lupini strain SH23  در تصفیه آب و پساب آلوده به هیدروکربن‌های نفتی و فلزات سنگین استفاده کرد.

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Determining sensitivity to heavy metals in surfactant-producing bacteria and their efficiency in removing Total petroleum hydrocarbons

نویسندگان [English]

  • Sahar Karimi 1
  • Shayan Shariati 2
  • Ahmad Ali pourbabaei 1
  • Hossein Ali Alikhani 3
  • Reyhaneh Kalami 1
1 Department of Soil Science Engineering, University of Tehran, Karaj, Iran.
2 Department of Environmental Engineering, Faculty of Environment, University of Tehran, Tehran, Iran.
3 Department of Soil Science, College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran.
چکیده [English]

Due to the association of some heavy metals with petroleum hydrocarbons, the use of native degrading bacteria of surfactant-producing oil that is resistant to heavy metals is one of the priorities of bioremediation technology. For this purpose, in this study, 39 alkane-degrading and biosurfactant-producing strains were used (isolated from saline soils contaminated with oil in the oil-rich areas of southern Khuzestan province) belonging to the soil microbiology laboratory gene bank of Tehran University. Then, the sensitivity of the isolates to different concentrations (40 to 80 ppm) of heavy metals (chromium, lead, copper, nickel, and cadmium) was tested. The results showed that only three strains Ochrobactrum lupini strain SH23, Ochrobactrum lupini strain SH24, and Bacillus subtilis subsp. Inaquosorum strain SH34 could grow in the presence of all heavy metals. The effects of different concentrations of heavy metals on the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of selected bacteria showed that Ochrobactrum lupini strain SH23 had the highest tolerance to heavy metals. The evaluation of the ability to degrade petroleum hydrocarbons in the presence of heavy metals by selected bacteria showed that the microbial consortium (SH23, SH24, and SH34) and Ochrobactrum lupini strain SH23 were able to degrade 71 and 64% of crude oil with a concentration of 0.5% (v/v), respectively after 10 days of aeration at 30 0C. The findings of this research proved that by conducting additional environmental studies, the bacterium Ochrobactrum lupini strain SH23 can be used in the treatment of water and wastewater contaminated with petroleum hydrocarbons and heavy metals.

کلیدواژه‌ها [English]

  • Biosurfactant
  • Diesel
  • Heavy metals tolerant bacteria
  • Total Petroleum hydrocarbons

Determining sensitivity to heavy metals in surfactant-producing bacteria and their efficiency in removing Total petroleum hydrocarbons

EXTENDED ABSTRACT

Introduction

Petroleum hydrocarbons with a variety of derivatives and varying degrees of toxicity, along with heavy metals, are the most important group of pollutants from oil entering the soil and aquatic environments, which have caused many problems for the environment. Owing to the co-contamination of heavy metals and total petroleum hydrocarbons in the ecosystem, the aim of this study was the isolation and application of native degrading bacteria of surfactant-producing oil and resistance to heavy metals in the degradation of total petroleum hydrocarbon along with heavy metals.

Materials and Methods

Thirty-nine bacterial strains with the ability to break down alkanes and produce biosurfactants (isolated from 10 oil-contaminated saline soils in the oil-rich areas of southern Khuzestan province) were selected from the microorganism gene bank of the soil microbiology laboratory of the University of Tehran.

To investigate the sensitivity of the strains to heavy metals, the growth rate of each strain (inoculation: 107 bacteria/mL and 1% v/v) was separately measured by the Spectrophotometer (OD600) in the presence of heavy metals on condition 72 h incubation,  36-38 C0, and 120 rpm. After selecting the best strains, the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of heavy metals chromium, copper, nickel, lead, and cadmium were determined according to the Clinical & Laboratory Standard Institute protocol. Then, the rate of degradation of 0.5% (v/v) crude oil by superior strains and their consortium (1% v/v, OD600 = 0.5) was investigated in the presence of heavy metals with three replication during 10 days of incubation, 38 C0, and 120 rpm. The growth rate of bacteria was measured using a spectrophotometer (USA, Unico 2100) at a wavelength of 600 nm (OD600). To measure the amount of crude oil degraded, 10 ml of the culture media was centrifuged at 6000 rpm for 20 minutes to precipitate the bacterial cells. Then, an equal volume of n-hexane was added to the solution as a solvent. Afterward, the extraction was performed by a separatory funnel. Then, the turbidity of the organic phase was measured by a spectrophotometer at a wavelength of 420 nm (OD420).

Results and Discussion

The results of the sensitivity of 39 bacterial strains to heavy metals showed that about 10% of the isolates were sensitive to all heavy metal and the highest level of sensitivity to metals was for cadmium (20%). Among the 39 bacterial strains, only three isolates Ochrobactrum lupini SH23, Ochrobactrum lupini SH24, and Bacillus subtilis subsp. Inaquosorum SH34 showed the ability to grow in media containing all heavy metals. The results of the effect of different concentrations of heavy metals on MBC and MIC of selected bacteria showed that Ochrobactrum lupini SH23 had the highest tolerance to heavy metals. The results of the crude oil degradation test in the presence of heavy metals showed that the consortium of selected bacteria, Ochrobactrum lupini SH23, Ochrobactrum lupini SH24, and Bacillus subtilis subsp. Inaquosorum SH34 were able to degrade 71%, 64%, 27%, and 22% of crude oil respectively, at 10 d on incubation (120 rpm, 38 C0).

Conclusion

Overall, the results showed that the new bacterial strains isolated from soils contaminated with a mixture of pollutants may have advanced catabolic capabilities and the ability to tolerate extreme conditions of toxic metals and nutrient limitations. Ochrobactrum lupini SH23 had a high ability in biosurfactant production, resistance to heavy metals (lead, chromium, cadmium, nickel, and copper), and removal of total petroleum hydrocarbons in the presence of heavy metal in the liquid media. Based on our findings, the bacterium Ochrobactrum lupini strain SH23 can be used in the treatment of water and wastewater contaminated with petroleum hydrocarbons and heavy metals.

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تحقیقات آب و خاک ایران
دوره 54، شماره 10
دی 1402
صفحه 1565-1579

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