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

Document Type : Research Paper

Authors

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 Engineering, University of Tehran, Karaj, Iran

4 Department of Soil Science, College of Agriculture & Natural Resources, University of Tehran, Karaj, Iran.

Abstract

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.

Keywords

Main Subjects

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.

References

Abolahrar, S., Kafilzadeh, F., & Kargar, M. (2014). Identifying cadmium resistant bacteria and evaluating their resistance spectrum during quarterly monitoring on the Kor River. Journal of Environmental Science and Technology, 16 (3), 165-179.
Abou-Shanab, R. A., Eraky, M., Haddad, A. M., Abdel-Gaffar, A. R. B., & Salem, A. M. (2016). Characterization of crude oil degrading bacteria isolated from contaminated soils surrounding gas stations. Bulletin of environmental contamination and toxicology, 97, 684-688.
Al-Mur, B. A., Pugazhendi, A., & Jamal, M. T. (2021). Application of integrated extremophilic (halo-alkalo-thermophilic) bacterial consortium in the degradation of petroleum hydrocarbons and treatment of petroleum refinery wastewater under extreme condition. Journal of Hazardous Materials, 413, 125351.
Aljerf, L., & Choukaife, A. E. (2018). Review: assessment of the doable utilisation of dendrochronology as an element tracer technology in soils artificially contaminated with heavy metals. Biodiversity Int J, 2 (1), 00037.
Angle, J. S., & Chaney, R. L. (1989). Cadmium resistance screening in nitrilotriacetate-buffered minimal media. Applied and environmental microbiology, 55 (8), 2101-2104.
Arulazhagan, P., & Vasudevan, N. (2011). Biodegradation of polycyclic aromatic hydrocarbons by a halotolerant bacterial strain Ochrobactrum sp. VA1. Marine pollution bulletin, 62 (2), 388-394.
Behdarvandan, P., Jalilzadeh Yengejeh, R., Sabzalipour, S., Roomiani, L., & Payandeh, K. (2020). Bioremediation of crude oil by indigenous species isolated from oil sludge contaminated soil. A case study: Karun Gas Oil Production Company,(IRAN). Journal of Advances in Environmental Health Research, 8 (4), 234-241.
Bezza, F. A., Beukes, M., & Chirwa, E. M. N. (2015). Application of biosurfactant produced by Ochrobactrum intermedium CN3 for enhancing petroleum sludge bioremediation. Process biochemistry, 50 (11), 1911-1922.
Bhattacharya, M., Biswas, D., Sana, S., & Datta, S. (2015). Biodegradation of waste lubricants by a newly isolated Ochrobactrum sp. C1. 3 Biotech, 5, 807-817.
Bhattacharya, M., Biswas, D., Sana, S., & Datta, S. (2014). Utilization of waste engine oil by Ochrobactrum pseudintermedium strain C1 that secretes an exopolysaccharide as a bioemulsifier. Biocatalysis and Agricultural Biotechnology, 3 (4), 167-176.
Bruins, M. R., Kapil, S., & Oehme, F. W. (2000). Microbial resistance to metals in the environment. Ecotoxicology and environmental safety, 45 (3), 198-207.
Cerqueda-García, D., García-Maldonado, J. Q., Aguirre-Macedo, L., & García-Cruz, U. (2020). A succession of marine bacterial communities in batch reactor experiments during the degradation of five different petroleum types. Marine Pollution Bulletin, 150, 110775.
Chen, C., Lei, W., Lu, M., Zhang, J., Zhang, Z., Luo, C., ... & Shen, Z. (2016). Characterization of Cu (II) and Cd (II) resistance mechanisms in Sphingobium sp. PHE-SPH and Ochrobactrum sp. PHE-OCH and their potential application in the bioremediation of heavy metal-phenanthrene co-contaminated sites. Environmental Science and Pollution Research, 23, 6861-6872.
Chen, W., Kong, Y., Li, J., Sun, Y., Min, J., & Hu, X. (2020). Enhanced biodegradation of crude oil by constructed bacterial consortium comprising salt-tolerant petroleum degraders and biosurfactant producers. International Biodeterioration & Biodegradation, 154, 105047.
Chodak, M., Gołębiewski, M., Morawska-Płoskonka, J., Kuduk, K., & Niklińska, M. (2013). Diversity of microorganisms from forest soils differently polluted with heavy metals. Applied Soil Ecology, 64, 7-14.
Bong, C. W., Malfatti, F., Azam, F., Obayashi, Y., & Suzuki, S. (2010). The effect of zinc exposure on the bacteria abundance and proteolytic activity in seawater. Interdisciplinary studies on environmental chemistry—biological responses to contaminants. Terrapub, Tokyo, 57-63.
Chunyan, X., Qaria, M. A., Qi, X., & Daochen, Z. (2023). The role of microorganisms in petroleum degradation: Current development and prospects. Science of The Total Environment, 865, 161112.
Cui, J. Q., He, Q. S., Liu, M. H., Chen, H., Sun, M. B., & Wen, J. P. (2020). Comparative study on different remediation strategies applied in petroleum-contaminated soils. International journal of environmental research and public health, 17 (5), 1606.
Eraky, M., Abou-Shanab, R. A. I., Salem, A. M., & Abdelgaffer, A. R. B. (2015). Petroleum hydrocarbon degradation potential of Ochrobactrum lupini isolated from BTEX enrichment soil. International Journal of Environment. 4 (3): 204-209.
Gonzalez Henao, S., & Ghneim-Herrera, T. (2021). Heavy metals in soils and the remediation potential of bacteria associated with the plant microbiome. Frontiers in Environmental Science, 15.
Hemmat-Jou, M. H., Safari-Sinegani, A. A., Mirzaie-Asl, A., & Tahmourespour, A. A. (2018). Analysis of microbial communities in heavy metals-contaminated soils using the metagenomic approach. Ecotoxicology, 27, 1281-1291.
Ibrahim, H. M. (2016). Biodegradation of used engine oil by novel strains of Ochrobactrum anthropi HM-1 and Citrobacter freundii HM-2 isolated from oil-contaminated soil. 3 Biotech, 6 (2), 226.
Isaac, P., Martínez, F. L., Bourguignon, N., Sánchez, L. A., & Ferrero, M. A. (2015). Improved PAHs removal performance by a defined bacterial consortium of indigenous Pseudomonas and actinobacteria from Patagonia, Argentina. International Biodeterioration & Biodegradation, 101, 23-31.
Jadoon, S., & Malik, A. D. N. A. (2017). DNA damage by heavy metals in animals and human beings: an overview. Biochem Pharmacol, 6 (3), 1-8.
Jaishankar, M., Tseten, T., Anbalagan, N., Mathew, B. B., & Beeregowda, K. N. (2014). Toxicity, mechanism and health effects of some heavy metals. Interdisciplinary toxicology, 7 (2), 60.
Kalami, R., & Pourbabaee, A.A. (2021). Investigating the potential of bioremediation in aged oil-polluted hypersaline soils in the south oilfields of Iran. Environmental Monitoring and Assessment. 193 (8), 517. doi: 10.1007/s10661-021-09304-7. PMID: 34309727.
Kalvandi, S., Garousin, H., Pourbabaee, A. A., & Alikhani, H. A. (2022a). Formulation of a culture medium to optimize the production of lipopeptide biosurfactant by a new isolate of Bacillus sp.: a soil heavy metal mitigation approach. Frontiers in Microbiology, 214.
Kalvandi, S., Garousin, H., Pourbabaee, A. A., & Farahbakhsh, M. (2022b). The release of petroleum hydrocarbons from a saline-sodic soil by the new biosurfactant-producing strain of Bacillus sp. Scientific Reports, 12 (1), 19770.
Kumar, C. G., Sujitha, P., Mamidyala, S. K., Usharani, P., Das, B., & Reddy, C. R. (2014). Ochrosin, a new biosurfactant produced by halophilic Ochrobactrum sp. strain BS-206 (MTCC 5720): purification, characterization and its biological evaluation. Process Biochemistry, 49 (10), 1708-1717.
Li, X., Meng, D., Li, J., Yin, H., Liu, H., Liu, X., ... & Yan, M. (2017). Response of soil microbial communities and microbial interactions to long-term heavy metal contamination. Environmental Pollution, 231, 908-917.
Li, Q., You, P., Hu, Q., Leng, B., Wang, J., Chen, J., ... & Ouyang, K. (2020). Effects of co-contamination of heavy metals and total petroleum hydrocarbons on soil bacterial community and function network reconstitution. Ecotoxicology and Environmental Safety, 204, 111083.
Margesin, R., Płaza, G. A., & Kasenbacher, S. (2011). Characterization of bacterial communities at heavy-metal-contaminated sites. Chemosphere, 82 (11), 1583-1588.
Ortega-González, D. K., Cristiani-Urbina, E., Flores-Ortíz, C. M., Cruz-Maya, J. A., Cancino-Díaz, J. C., & Jan-Roblero, J. (2015). Evaluation of the removal of pyrene and fluoranthene by Ochrobactrum anthropi, Fusarium sp. and their coculture. Applied biochemistry and biotechnology, 175, 1123-1138.
Peng, H., Xie, W., Li, D., Wu, M., Zhang, Y., Xu, H., ... & Liu, W. (2019). Copper-resistant mechanism of Ochrobactrum MT180101 and its application in membrane bioreactor for treating electroplating wastewater. Ecotoxicology and Environmental Safety, 168, 17-26.
Poszytek, K., Karczewska-Golec, J., Ciok, A., Decewicz, P., Dziurzynski, M., Gorecki, A., ... & Dziewit, L. (2018). Genome-guided characterization of Ochrobactrum sp. POC9 enhancing sewage sludge utilization—Biotechnological potential and biosafety considerations. International Journal of Environmental Research and Public Health, 15 (7), 1501.
Primeia, S., Inoue, C., & Chien, M. F. (2020). Potential of biosurfactants’ production on degrading heavy oil by bacterial consortia obtained from tsunami-induced oil-spilled beach areas in Miyagi, Japan. Journal of Marine Science and Engineering, 8 (8), 577.
Pugazhendi, A., Qari, H., Basahi, J. M. A. B., Godon, J. J., & Dhavamani, J. (2017). Role of a halothermophilic bacterial consortium for the biodegradation of PAHs and the treatment of petroleum wastewater at extreme conditions. International Biodeterioration & Biodegradation, 121, 44-54.
Rahman, K.S.M., Thahira-Rahman, J., Lakshmanaperumalsamy, P., & Banat, IM. 2002. Towards efficient crude oil degradation by a mixed bacterial consortium. Bioresource technology, 85(3), pp.257-261.
Rashid, M. I., Mujawar, L. H., Shahzad, T., Almeelbi, T., Ismail, I. M., & Oves, M. (2016). Bacteria and fungi can contribute to nutrients bioavailability and aggregate formation in degraded soils. Microbiological research, 183, 26-41.
Ron, E. Z., & Rosenberg, E. (2014). Enhanced bioremediation of oil spills in the sea. Current Opinion in biotechnology, 27, 191-194.
Roszak, M., Jabłońska, J., Stachurska, X., Dubrowska, K., Kajdanowicz, J., Gołębiewska, M., ... & Karakulska, J. (2021). Development of an autochthonous microbial consortium for enhanced bioremediation of PAH-contaminated soil. International Journal of Molecular Sciences, 22 (24), 13469.
Safahieh, A., Lamoochi, R., Salamat, N., & Abyar, H. (2014) Isolation and Identification of Bacillus firmus from the Marine Sediments of Imam Khomeini Port and Study Its Ability in Biosorption of Lead. Journal of Oceanography, 5 (17):11-19. (In Persian)
Shahaliyan, F., Safahieh, A. R., & Abyar, H. (2013). Isolation and Identification of Nickel Resistant Bacteria in Khor Mousa Sediments and Study of Bacterial Capability in Nickel Biosorption. Journal of Environmental Studies, 39 (2), 93-100. (In Persian).
 Sharma, B., & Shukla, P. (2021). A comparative analysis of heavy metal bioaccumulation and functional gene annotation towards multiple metal resistant potential by Ochrobactrum intermedium BPS-20 and Ochrobactrum ciceri BPS-26. Bioresource Technology, 320, 124330.
Sales da Silva, I. G., Gomes de Almeida, F. C., Padilha da Rocha e Silva, N. M., Casazza, A. A., Converti, A., & Asfora Sarubbo, L. (2020). Soil bioremediation: Overview of technologies and trends. Energies, 13 (18), 4664.
Singh, S., Parihar, P., Singh, R., Singh, V. P., & Prasad, S. M. (2016). Heavy metal tolerance in plants: role of transcriptomics, proteomics, metabolomics, and ionomics. Frontiers in plant science, 6, 1143.
Slepecky R.A, Hemphill H.E. The Prokaryotes, 3rd Ed.. New York: Springer; 2006
Syed, S., & Chinthala, P. (2015). Heavy metal detoxification by different Bacillus species isolated from solar salterns. Scientifica, 2015 (Article ID 319760):1- 8.
Tiwari, B., Manickam, N., Kumari, S., & Tiwari, A. (2016). Biodegradation and dissolution of polyaromatic hydrocarbons by Stenotrophomonas sp. Bioresource Technology, 216, 1102-1105.
Villagrasa, E., Palet, C., López-Gómez, I., Gutiérrez, D., Esteve, I., Sánchez-Chardi, A., & Solé, A. (2021). Cellular strategies against metal exposure and metal localization patterns linked to phosphorus pathways in Ochrobactrum anthropi DE2010. Journal of Hazardous Materials, 402, 123808.
Xue, J., Yu, Y., Bai, Y., Wang, L., & Wu, Y. (2015). Marine oil-degrading microorganisms and biodegradation process of petroleum hydrocarbon in marine environments: a review. Current microbiology, 71, 220-228.
Wang, S., Wang, D., Yu, Z., Dong, X., Liu, S., Cui, H., & Sun, B. (2021). Advances in research on petroleum biodegradability in soil. Environmental Science: Processes & Impacts, 23 (1), 9-27.
Yang, Y., Wang, J., Liao, J., Xie, S., & Huang, Y. (2015). Abundance and diversity of soil petroleum hydrocarbon-degrading microbial communities in oil exploring areas. Applied microbiology and biotechnology, 99, 1935-1946.
Zhang, X., Zhang, Q., Yan, T., Jiang, Z., Zhang, X., & Zuo, Y. Y. (2015). Quantitatively predicting bacterial adhesion using surface free energy determined with a spectrophotometric method. Environmental science & technology, 49 (10), 6164-6171.
Iranian Journal of Soil and Water Research
Volume 54, Issue 10
January 2024
Pages 1565-1579

Files

Share

How to cite

Statistics