Welcome to Bilingual Publishing Group

The Efficiency of Integrated Bioremediation Techniques in Restoring Oil-Contaminated Soils

Authors

  • Samira Nadjafova ORCID Department of Bioecology,Baku State University,Baku AZ1148,Azerbaijan
  • Mahluga Yusifova Email ORCID Department of Geographical Ecology,Baku State University,Baku AZ1148,Azerbaijan
  • Cinara Bagirova ORCID Department of Biology,Sumgait State University,Sumgait AZ5008,Azerbaijan
  • Aygun Gasimova ORCID Department of Soil and Water Microbiology,Institute of Molecular Biology,Ministry of Science and Education of the Republic ofAzerbaijan,Baku AZ1073,Azerbaijan
  • Tursynkul Bazarbayeva ORCID UNESCO Chair in Sustainable Development,Al-Farabi Kazakh National University,Almaty 050040,Kazakhstan
  • Sona Osmanova ORCID Department of Geographical Ecology,Baku State University,Baku AZ1148,Azerbaijan
  • Ramil Sadigov ORCID Department of Engineering and Applied Sciences,Azerbaijan State University of Economics,Baku AZ1001,Azerbaijan
  • Kamala Nuriyeva ORCID Department of Agroecology,Institute of Geography named after Academician H.A.Aliyev,Ministry of Science and Education of the Republic ofAzerbaijan,Baku AZ1143,Azerbaijan
  • Matanat Aliyeva ORCID Department of Agroecology,Institute of Geography named after Academician H.A.Aliyev,Ministry of Science and Education of the Republic ofAzerbaijan,Baku AZ1143,Azerbaijan
  • Nazila Alverdiyeva ORCID Department of Engineering and Applied Sciences,Azerbaijan State University of Economics,Baku AZ1001,Azerbaijan

DOI:

https://doi.org/10.30564/re.v8i5.117312

Received:16 August 2026 |Revised:18 September 2026 |Accepted:24 September 2026 |Published Online:12 October 2026

Abstract

Contamination of soil ecosystems with oil and petroleum products remains a key environmental problem, requiring the development of effective and environmentally friendly restoration methods. Bioremediation using degradative microorganisms and plants is considered the most promising method for restoring man-made lands. This article examines the effectiveness of the integrated, phased application of chemical, biotechnological, and phytomeliorative techniques for restoring oil-contaminated gray-brown soils (Gypsisols) on the Absheron Peninsula (Azerbaijan), covering approximately 25,000 ha. A laboratory model experiment on the bioremediation of oil-contaminated soils was conducted over 180 days using various ameliorant treatments and native oil-degrading bacteria isolated from these soils and tested for their destructive activity. Microbial counts were determined by plating on nutrient media. The degree of oil biodegradation was monitored using Expand to United States Environmental Protection Agency (EPA) method 3540, and the hydrocarbon-degrading activity of the strains was determined by optical density. Phytotoxicity before and after the experiment was assessed according to ISO 11269-2:2012. The effectiveness of phytoremediation using garden pea (Pisum sativum L.) was assessed based on the total microbial count and hydrocarbon loss in the soil. Laboratory modeling results showed that improving substrate properties with dolomite flour created favorable conditions for the development of an introduced consortium of native bacteria (Arthrobacter sp., Bacillus sp., Pseudomonas sp.), while the final use of phytoremediation (Pisum sativum L.) stimulated microbial community development through the rhizosphere effect. Thus, the integrated approach contributed to a 95–97% reduction in the initial amount of petroleum hydrocarbons within 6 months. These results confirm the high effectiveness of the integrated application of various techniques for the environmentally safe restoration of industrially oriented soils in arid climates, opening up prospects for the reclamation of disturbed lands.

Keywords:

Soils,Oil Pollution,Dolomite Flour,Hydrocarbon-Oxidizing Microorganisms,Phytomeliorants,Self-Purification

References

[1] Ossai, I.C., Ahmed, A., Hassan, A., et al., 2020. Remediation of soil and water contaminated with petroleum hydrocarbon: A review. Environmental Technology & Innovation. 17, 100526. DOI: https://doi.org/10.1016/j.eti.2019.100526

[2] Ambaye, T.G., Chebbi, A., Formicola, F., et al., 2022. Ex-situ bioremediation of petroleum hydrocarbon contaminated soil using mixed stimulants: Response and dynamics of bacterial community and phytotoxicity. Journal of Environmental Chemical Engineering. 10(6), 108814. DOI: https://doi.org/10.1016/j.jece.2022.108814

[3] Gao, H., Wu, M., Liu, H., et al., 2022. Effect of petroleum hydrocarbon pollution levels on the soil microecosystem and ecological function. Environmental Pollution. 293, 118511. DOI: https://doi.org/10.1016/j.envpol.2021.118511

[4] Mammadov, G.Sh., Mammadova, S.Z., Yusifova, M.M., et al., 2025. Soil fertility management in the Gusar-Gonagkend cadastral district of Azerbaijan based on agroecological land assessment. Advances in Biology & Earth Sciences. 10(2), 226–244. DOI: https://doi.org/10.62476/abes.102226

[5] Koshlaf, E., Ball, A.S., 2017. Soil bioremediation approaches for petroleum hydrocarbon polluted environments. AIMS Microbiology. 3(1), 25–49. DOI: https://doi.org/10.3934/microbiol.2017.1.25

[6] Ismailov, N.M., Nadjafova, S.I., 2017. Landscape Stability along the Main Export Pipeline Baku–Tbilisi–Ceyhan (Baku–Supsa) against Crude Oil Contamination. INFRA-M: Moscow, Russia. Available from: https://www.preslib.az/ru/book/2ci58qiguX2paq3 (in Russian)

[7] Filatov, D.A., Ovsyannikova, V.S., 2017. Natural processes of self-restoration of oil-contaminated biocenoses. Ecological Bulletin of Russia. 10, 17–20. (in Russian)

[8] Das, N., Chandran, P., 2011. Microbial degradation of petroleum hydrocarbon contaminants: An overview. Biotechnology Research International. 2011(1), 941810. DOI: https://doi.org/10.4061/2011/941810

[9] Roszak, M., Jabłońska, J., Stachurska, X., et al., 2021. Development of an autochthonous microbial consortium for enhanced bioremediation of PAH-contaminated soil. International Journal of Molecular Sciences. 22(24), 13469. DOI: https://doi.org/10.3390/ijms222413469

[10] Kanaujia, P.K., Gupta, P., Dubey, A.K., et al., 2026. Microbial and plant-based bioremediation of petroleum hydrocarbons: Mechanisms, strategies and environmental applications. Journal of Environmental & Earth Sciences. 8(4), 66–87. DOI: https://doi.org/10.30564/jees.v8i4.13137

[11] El-Baz, A.F., Shetaia, Y.M., Abdelghani, D.Y., et al., 2025. Microbial bioremediation of contaminated soil. In: Shah, M.P., Deka, D. (Eds.). Soil Bioremediation. Springer: Cham, Switzerland. pp. 33–74. DOI: https://doi.org/10.1007/978-3-031-76886-6_2

[12] Natarajan, S., Subbiah, M., Manam, V.K., et al., 2024. Microbial-meditated remediation of crude oil-contaminated soil. In: Karnwal, A., Mohammad Said Al-Tawaha, A.R. (Eds.). Microbial Applications for Environmental Sustainability. Springer: Singapore. pp. 71–98. DOI: https://doi.org/10.1007/978-981-97-0676-1_4

[13] Barzegar, G., Jorfi, S., Sanaei, D., et al., 2025. Bioremediation of diesel-contaminated saline soil and enhancement of microbial salinity tolerance by a biosurfactant-producing Bacillus subtilis AHV-KH11 and external surfactant application: Bio-toxicity assessment. Scientific Reports. 15, 20673. DOI: https://doi.org/10.1038/s41598-025-06954-2

[14] Krishna Raj, C., Siranjeevi, R., Susmitha, R., et al., 2026. Bioremediation in action: Recent progress, challenges, and future directions for environmental restoration. Biodegradation. 37(1), 9. DOI: https://doi.org/10.1007/s10532-025-10232-3

[15] Baruah, N.P., Goswami, M., Sarma, N., et al., 2024. Pioneering technologies over time to rehabilitate crude oil-contaminated ecosystems: A review. Environmental Science and Pollution Research. 31(55), 63576–63602. DOI: https://doi.org/10.1007/s11356-024-35442-9

[16] Salum, J.A., Kapoor, B., Sharma, S., 2026. Genetically engineered organisms for sustainable bioremediation of polluted environments. Biodegradation. 37(2), 49. DOI: https://doi.org/10.1007/s10532-026-10268-z

[17] Elshafei, A.M., Mansour, R., 2024. Microbial bioremediation of soils contaminated with petroleum hydrocarbons. Discover Soil. 1, 9. DOI: https://doi.org/10.1007/s44378-024-00004-5

[18] Veluri, S., Somadri, V., Vaishnavi, V., et al., 2026. In situ and ex situ bioremediation techniques for removal of emerging pollutants in the environment. In: Reddy, K.V., Maddela, N.R., Reddy, A.V.K. (Eds.). Emerging Contaminants in the Environment. Springer: Cham, Switzerland. pp. 207–233. DOI: https://doi.org/10.1007/978-3-032-10432-8_10

[19] Sahith, V.N., Kumar, J.A., Sruthi, V.S., et al., 2026. Bioremediation of polycyclic aromatic hydrocarbons contaminated soils/water for environmental remediation. Biodegradation. 37(1), 3. DOI: https://doi.org/10.1007/s10532-025-10229-y

[20] Pinto, A.P.H., Marques, C.S., Carreiro, E.P., et al., 2026. Sustainable bioremediation techniques for soil petroleum hydrocarbons. In: Parnian, A., Vara Prasad, M.N., Furze, J.N., et al. (Eds.). Bioremediation of Petroleum Contaminated Soil and Sediment. Springer: Cham, Switzerland. pp. 123–169. DOI: https://doi.org/10.1007/978-3-031-95524-2_7

[21] Hashim, H.S., Zayan, M.M., Mohamed, A.A., 2026. Actinomycetes in the spotlight: Biodiversity and their role in bioremediation. World Journal of Microbiology and Biotechnology. 42, 44. DOI: https://doi.org/10.1007/s11274-025-04610-5

[22] Manickam, N., Arumugamurthi, S., Gopal, A., et al., 2025. Investigation of three traditional soil amendments for reviving soil health in petroleum-contaminated soils. Environmental Science and Pollution Research. 32(46), 26513–26523. DOI: https://doi.org/10.1007/s11356-025-37167-9

[23] Ruley, J.A., Tomor, B.M., 2024. Biostimulation of microbes for enhanced oil removal from petroleum hydrocarbon-contaminated soils: A zero waste remediation approach. In: Bhat, R.A., Dar, G.H., Hajam, Y.A. (Eds.). Zero Waste Management Technologies. Springer: Cham, Switzerland. pp. 323–341. DOI: https://doi.org/10.1007/978-3-031-57275-3_16

[24] Das, A., Das, N., Rajkumari, J., et al., 2024. Exploring the bioremediation potential of Bacillus spp. for sustainable mitigation of hydrocarbon contaminants. Environmental Sustainability. 7(2), 135–156. DOI: https://doi.org/10.1007/s42398-024-00309-9

[25] Okoye, A.U., Selvarajan, R., Chikere, C.B., et al., 2024. Characterization and identification of long-chain hydrocarbon-degrading bacterial communities in long-term chronically polluted soil in Ogoniland: An integrated approach using culture-dependent and independent methods. Environmental Science and Pollution Research. 31(21), 30867–30885. DOI: https://doi.org/10.1007/s11356-024-33326-6

[26] Melo, L.B.U., Borrego, B.B., Gracioso, L.H., et al., 2025. Microbial degradation of diesel fuel as a potential environmental bioremediation. Biodegradation. 36(5), 100. DOI: https://doi.org/10.1007/s10532-025-10194-6

[27] Khalili, B., Zhao, S., Khodadadi, N., et al., 2025. Using magnetic susceptibility for assessing total petroleum hydrocarbon biodegradation in Pseudomonas aeruginosa and Marinobacter hydrocarbonoclasticus-inoculated soils. Water, Air, & Soil Pollution. 236(7), 428. DOI: https://doi.org/10.1007/s11270-025-08091-y

[28] Soto-Mancilla, M., Fioroni, F., Ebrecht, L., et al., 2026. Fungal bioremediation of petroleum hydrocarbons in terrestrial environments: A systematic review and meta-analysis. Biodegradation. 37, 68. DOI: https://doi.org/10.1007/s10532-026-10292-z

[29] Vasilyeva, G.K., Strijakova, E.R., Ortega-Calvo, J.J., 2024. Remediation of soils polluted by oil industries. In: Ortega-Calvo, J.J., Coulon, F. (Eds.). Soil Remediation Science and Technology. Springer: Cham, Switzerland. pp. 191–234. DOI: https://doi.org/10.1007/698_2024_1080

[30] Kumar, S., Banerjee, S., Ghosh, A., et al., 2026. Biological synergies for heavy metal bioremediation across microbial, algal, and phytoremediation perspectives. Discover Green Chemistry. 1, 3. DOI: https://doi.org/10.1007/s44509-026-00003-5

[31] Ibrahim, S.Y., Abdelhamid, E.A., El-Hagrassi, A.M., et al., 2025. Statistical optimization of crude oil bioremediation using Streptomyces aurantiogriseus isolated from Egypt’s Western Desert. Biodegradation. 36(4), 68. DOI: https://doi.org/10.1007/s10532-025-10154-0

[32] Yu, J., Zhang, X., Zhang, Z., et al., 2025. Effects of bioremediation on soil fertility and microbial communities of degraded grassland soil under dual petroleum contamination and saline-alkali stress. Journal of Forestry Research. 36(1), 20. DOI: https://doi.org/10.1007/s11676-024-01809-4

[33] Hajiyeva, S., Aliyeva, T., Yusifova, M., 2020. The ecological state of Boyuk Shor lake of Azerbaijan. Environmental Monitoring and Assessment. 192(12), 780. DOI: https://doi.org/10.1007/s10661-020-08762-9

[34] Nadjafova, S.I., Bagirova, C.Z., 2021. Ways to improve biogeneity, assimilation potential and fertility of soil cover of the urban landscapes of Sumgait city. South of Russia: Ecology, Development. 16(3), 88–94. DOI: https://doi.org/10.18470/1992-1098-2021-3-88-94 (in Russian)

[35] Babayev, M.P., Ramazanova, F., Mirzazade, R., et al., 2023. Soils of the Azerbaijan Republic (Kur-Araz Valley, Ganja-Gazakh Inclination Plain and Irrigated Soils of the Absheron Peninsula and Their Productive Ability). Elm: Baku, Azerbaijan. Available from: https://books.google.com/books/about/Soils_of_the_Azerbaijan_Republic_Kur_Ara.html?id=ZFAa0gEACAAJ

[36] ISO 18400-102:2017. 2017. Soil Quality—Sampling—Part 102: Selection and Application of Sampling Techniques. Available from: https://www.iso.org/standard/62843.html

[37] US EPA SW-846 Method 3540C. 1996. Soxhlet Extraction. Available from: https://www.epa.gov/hw-sw846/sw-846-test-method-3540c-soxhlet-extraction

[38] Green, L.H., Goldman, E. (Eds.), 2021. Practical Handbook of Microbiology, 4th ed. CRC Press: Boca Raton, FL, USA. DOI: https://doi.org/10.1201/9781003099277

[39] ABIS, 2023. Advanced Bacterial Identification Software for Windows and Android. Regnum Prokaryotae Database. Available from: https://www.abisonline.eu/ABIS/bacteria_abis.html

[40] ISO 11269-2:2012. 2012. Soil Quality—Determination of the Effects of Pollutants on Soil Flora—Part 2: Effects of Contaminated Soil on the Emergence and Early Growth of Higher Plants. Available from: https://www.iso.org/standard/51382.html

[41] ISO 7218:2024. 2024. Microbiology of the Food Chain—General Requirements and Guidance for Microbiological Examinations. Available from: https://www.iso.org/standard/79508.html

[42] ISO 22174:2024. 2024. Microbiology of the Food Chain—Polymerase Chain Reaction (PCR) for the Detection and Quantification of Microorganisms—General Requirements and Definitions. Available from: https://www.iso.org/standard/81777.html

[43] Yolchueva, U.J., Jafarova, R.A., Azizbeyli, E.I., et al., 2017. Luminescent aromatic hydrocarbons of Balakhani oil of the Absheron Peninsula of Azerbaijan. In Proceedings of the II All-Russian Scientific Conference with International Participation “Actual Problems of Adsorption and Catalysis”, Plyos, Russia, 28–30 June 2017; pp. 158–159. (in Russian)

[44] Taki, H., Syutsubo, K., Mattison, R.G., et al., 2007. Identification and characterization of o-xylene-degrading Rhodococcus spp. which were dominant species in the remediation of o-xylene-contaminated soils. Biodegradation. 18(1), 17–26. DOI: https://doi.org/10.1007/s10532-005-9030-x

[45] Jeong, E., Hirai, M., Shoda, M., 2008. Removal of o-xylene using biofilter inoculated with Rhodococcus sp. BTO62. Journal of Hazardous Materials. 152(1), 140–147. DOI: https://doi.org/10.1016/j.jhazmat.2007.06.078

[46] National Hydrometeorology Service of Azerbaijan, 2024. Multi-Year Climate Baselines and Agro-Climatic Resources of the Absheron Peninsula. Ministry of Ecology and Natural Resources: Baku, Azerbaijan. Available from: https://meteo.az/index.php?ln=az&pg=65 (in Azerbaijani)

[47] de Santana Nazaré, P.G.B., de Menezes Neto, A.A., de Oliveira, L.A., et al., 2026. Metabolic specialization and synergy in alkane degradation by a microbial consortium: An integrated genomic and geochemical approach. Current Microbiology. 83(5), 261. DOI: https://doi.org/10.1007/s00284-026-04858-6

[48] Mahmud, M.S., Chong, K.P., 2022. Effects of liming on soil properties and its roles in increasing the productivity and profitability of the oil palm industry in Malaysia. Agriculture. 12(3), 322. DOI: https://doi.org/10.3390/agriculture12030322

[49] Singha, L.P., Pandey, P., 2021. Rhizosphere assisted bioengineering approaches for the mitigation of petroleum hydrocarbons contamination in soil. Critical Reviews in Biotechnology. 41(5), 749–766. DOI: https://doi.org/10.1080/07388551.2021.1888066

[50] Pysarenko, P., Samojlik, M., Taranenko, A., et al., 2022. Microbial remediation of petroleum-polluted soil. Agraarteadus: Journal of Agricultural Science. 33(2), 434–441. DOI: https://doi.org/10.15159/jas.22.30

[51] Vasilyeva, G., Mikhedova, E., Zinnatshina, L., et al., 2022. Use of natural sorbents for accelerated bioremediation of grey forest soil contaminated with crude oil. Science of the Total Environment. 850, 157952. DOI: https://doi.org/10.1016/j.scitotenv.2022.157952

Downloads

Issue

Article Type

Article