پیامد آلودگی نفتی طبیعی و طولانی‌مدت بر جمعیت میکروبی و فعالیت آنزیم اوره‌آز خاک

نوع مقاله : مقالات پژوهشی

نویسندگان

1 گروه علوم و مهندسی خاک، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران

2 گروه علوم و مهندسی خاک، دانشکده کشاورزی، دانشگاه رازی کرمانشاه، کرمانشاه، ایران

3 به‌نژادی و بیوتکنولوژی گیاهی، گروه به‌نژادی و بیوتکنولوژی گیاهی، دانشکده کشاورزی، دانشگاه تبریز، تبریز، ایران

4 گروه گیاه‌پزشکی، دانشکده کشاورزی، دانشگاه رازی کرمانشاه، کرمانشاه، ایران

چکیده

آلاینده‌های آلی از جمله ترکیبات نفتی، یک مشکل جهانی برای سلامت محیط‌زیست و موجودات زنده محسوب می‌شوند که می­توانند ویژگی­های فیزیکی، شیمیایی و زیستی خاک را تحت تأثیر قرار دهند. در این تحقیق، شاخص‌های زیستی از جمله جمعیت میکروبی و فعالیت آنزیم اوره­آز در خاک­های آلوده به نفت نفت‌-شهر کرمانشاه مورد توجه بود. به‌منظور بررسی اثرات آلودگی نفتی طولانی­مدت و طبیعی، 120 نمونه خاک آلوده با سطوح مختلف نفت؛ آلودگی شدید (H:High)، متوسط (M:Moderate) و کم (L: Low) از عمق 15-0 سانتی­متری از 4 منطقه مختلف تهیه­شد. پس از اندازه‌گیری ویژگی‌های فیزیکوشیمیایی خاک‌ها، شمارش میکروبی و اندازه­گیری فعالیت اوره­آز انجام­شد. برای تعیین جمعیت میکروبی کل و باکتری­های درگیر در تجزیه نفت، به‌ترتیب اقدام به شمارش میکروبی در محیط کشت‌های NA (Nutrient Agar) و CFMM (Carbon Free Minimal Medium) شد که رابطه مستقیمی با افزایش میزان نفت داشت. میانگین درصد نفت اندازه‌گیری شده به روش سوکسله، به‌ترتیب 03/4، 95/9 و 50/22 درصد به‌ترتیب برای سطوح L، M و H به‌دست آمد. نتایج نشان داد که با افزایش شدت آلودگی، جمعیت میکروبی افزایش یافت. بالاترین جمعیت میکروبی شمارش­شده در محیط کشت NA، در خاک­های با آلودگی شدید CFU/g 105×54/9 و پایین­ترین جمعیت در خاک­های با آلودگی کمCFU/g  105×25/3 به‌دست آمد. در محیط کشت CFMM نیز بیشترین و کمترین جمعیت به‌ترتیب در خاک­های با آلودگی شدید و کم با مقادیر CFU/g 105×11/3 و 104×11/8 به‌دست آمد. میزان افزایش جمعیت میکروبی در دو محیط NA و CFMM با افزایش آلودگی به‌ترتیب 9/2 و 8/3 برابر بود. برای هردو محیط کشت، منطقه 1 دارای بیشترین جمعیت و بیشترین درصد آلودگی نفتی و منطقه 4 دارای کمترین جمعیت و کمترین درصد آلودگی نفتی بود. پایین­ترین فعالیت اوره­آز در منطقه 1 و بالاترین آن در منطقه 4 مشاهده شد. آلودگی نفتی نمونه­های خاک منجر به کاهش فعالیت اوره­آز شد به گونه­ای که بیشترین فعالیت آنزیمی در خاک­های با آلودگی کم ( µgNH4/g.h90/594) و کمترین فعالیت در خاک­های با آلودگی شدید (µgNH4/g.h 11/176) به‌دست آمد، میزان درصد کاهش فعالیت با افزایش سطح آلودگی 5/70 درصد بود. آنالیز مؤلفه­های اصلی نیز انجام شد و 71 درصد از واریانس تراکمی نمونه­ها توسط دو مؤلفه اول (مؤلفه بیوشیمیایی و مؤلفه فیزیکی) قابل توجیه بود. یافته‌های این تحقیق نشان داد که آلودگی نفتی طولانی­مدت و طبیعی باعث گزینش جامعه میکروبی مقاوم به نفت شده و بنابراین پاسخ مثبت آنها به حضور ترکیبات نفتی را شاهد بودیم اما فعالیت آنزیم اوره­آز تحت تأثیر آلودگی نفتی کاهش یافت. به‌نظر اثرات بازدارندگی ترکیبات نفتی بر فعالیت اوره‌از یا جامعه غالب میکروبی با فعالیت اوره‌از محدود، سبب شده است تا فعالیت اوره‌از واکنش منفی به حضور آلاینده نفتی نشان دهد.

کلیدواژه‌ها

موضوعات


عنوان مقاله [English]

Consequence of Natural and Long Term Oil Pollution on Microbial Population and Urease Activity of Soil

نویسندگان [English]

  • Shokufeh Moradi 1
  • Mohammad Reza Sarikhani 1
  • Ali Beheshti Ale Agha 2
  • Adel Reyhanitabarَ 1
  • Seyed Siamak Alavi-kia 3
  • Ali Bandehagh 3
  • Rouhallah Sharifi 4
1 Department of Soil Science, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
2 Department of Soil Science, Faculty of Agriculture, Razi University, Kermanshah, Iran
3 Department of Plant Breeding and Biotechnology, Faculty of Agriculture, University of Tabriz, Tabriz, Iran
4 Department of Plant Pathology, Faculty of Agriculture, Razi University, Kermanshah, Iran
چکیده [English]

Introduction
Oil contamination affects the biological, physical, and chemical properties of soil. The abundance and diversity of soil microbial communities can significantly be influenced by petroleum hydrocarbons. Soil biological indicators including microbial population and enzyme activity, are highly sensitive to environmental stresses and respond to them quickly. Measuring the microbial population is one of the most common biological indicators which is used to study the quality and health of the soil. Also, measuring the activity of enzymes such as urease is one of the most sensitive indicators of oil-contaminated soils. There are some studies on the effects of oil contamination on microbial population and soil enzyme activity. Most of the studies have tested non-natural and short-term oil pollution and reported the adverse effects of oil hydrocarbons on microbial activities in soil. While the soil sample used in this research had natural and long-term contamination and the microorganisms are compatible with polluted conditions. The aim of this study was to investigate changes in the microbial population and urease activity in the presence of different levels of oil contamination, and how petroleum hydrocarbons can affect them. Petroleum hydrocarbons are toxic and persistent in soil, so it is necessary to study the pattern of changes in soil biological characteristics in effective soil management.
 
Material and Methods
In this study, 120 samples of oil-contaminated soils were collected from the oil-rich area of Naft-Shahr (located in the west of Kermanshah province) which had natural and long-term oil pollution. A nested design was used to analysis data in this research. The test factors included locations (4 locations) and 3 different levels of oil pollution: low (L), moderate (M), and high (H). Also, 10 replications were considered in the three levels of oil contamination. The collected soils were analyzed for physico-chemical (pH, EC, Ɵm, CCE, OC, soil texture) and biological properties (including urease activity, BR and SIR) using standard methods, and the concentration of oil pollutants was determined by the Soxhlet extractor. To determine the abundance of the culturable microbial population, bacterial counting was performed using nutrient agar (NA) and carbon-free minimal medium (CFMM) supplemented with crude oil as the media. Urease activity was measured by the indophenol blue method and finally, the results of measuring chemical, physical and biological properties were analyzed using principal component analysis (PCA).
 
Results and Discussion
 The average percentage of oil measured by Soxhlet method was 4.03%, 9.95% and 22.50% respectively for L, M and H levels. The results showed that the microbial population increased with the increase of contamination intensity. The highest microbial population counted in NA culture medium was 9.54 ×105 CFU/g in H soils and the lowest population was 3.25 × 105 CFU/g in L soils. In the CFMM culture medium, the highest population in H soils was 11.3 × 105 CFU/g and the lowest population in L soils was 11.8 × 104 CFU/g. For both NA and CFMM mediums, location 1 had the highest population and location 4 had the lowest microbial population. Oil contamination of soil samples led to a decrease in urease activity in such a way that the highest enzyme activity in soils was obtained with low contamination (594.90 µgNH4/g.h) and the lowest activity in heavily contaminated soils (176.11 µgNH4/g.h). Also, the lowest urease activity was observed in location 1 and the highest in location 4. Principal components analysis (PCA) was also performed and 71% of the variance of the samples could be explained by the first two components (biochemical component and physical component). The results of this research indicated an increase in the microbial population with an increasing of the intensity of oil pollution. It seems that the results obtained from the studies conducted on man-made pollution and natural pollution have differences in terms of the type of biological responses. Aged, long-term and natural oil pollution has caused the selection of oil-resistant microbial community, and therefore we see their positive response to the presence of oil compounds. Conversely, urease enzyme activity was found to be higher in soils with low pollution. This suggests that microbial activity, while influential, is not the sole determinant of urease activity, and various factors contribute to Soil Enzyme Activity (SEA). The type of petroleum pollutant, the direct effect of petroleum compounds on urease-producing microorganisms, as well as the non-microbial origin of urease in soil can be possible reasons for reducing urease activity in contaminated soils.
 
Conclusion
In areas where petroleum pollutants are naturally and long-term present in the soil, some oil-decomposing microbial groups use petroleum hydrocarbons as a source of carbon for their nutrition, so the abundance of oil-decomposing communities increases. The results showed an increase in the microbial population with an increase in the intensity of oil pollution. On the other hand, the activity of urease enzyme measured in soils with low pollution was higher because non-microbial factors may affect the activity of this enzyme and the increase in the microbial population is not related to the increase in the population of urease-producing microbes.

کلیدواژه‌ها [English]

  • Biological indicators
  • Enzyme activity
  • Microbial population
  • Oil pollution

©2023 The author(s). This is an open access article distributed under Creative Commons Attribution 4.0 International License (CC BY 4.0), which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source.

  1.  

    1. Andreoni, V., Cavalca, L., Rao, M. A., Nocerino, G., Bernasconi, S., Dell’Amico, E., ... & Gianfreda, L. (2004). Bacterial communities and enzyme activities of PAHs polluted soils. Chemosphere, 57(5), 401-412. https://doi.org/10.1016/j.chemosphere.2004.06.013
    2. Christopher, S., Hein, P., Marsden, J., & Shurleff, A.S. (1988). Evaluation of methods 3540 (soxhlet) and 3550 (Sonication) for evaluation of appendix IX analyses from solid samples. S-CUBED, Report for EPA contract 68-03-33-75, work assignment No. 03, Document No (pp. 523-546). SSS.
    3. Cox, J.F., Blackstone, J.H., & Schleier, J.G. (2003). Managing operations: A focus on excellence. North River Press.
    4. Deaker, R., Kecskés, M.L., Rose, M.T., Amprayn, K., Ganisan, K., Tran, T.K.C., Vu, T.N., Phan, T.C., Nguyen, T.H., & Kennedy, I.R. (2011). Practical methods for the quality control of inoculant biofertilisers. Australian Centre for International Agricultural Research (ACIAR). 101 pp
    5. Dos Santos, H. F., Cury, J.C., Do Carmo, F.L., Dos Santos, A.L., Tiedje, J., van Elsas, J.D., ... & Peixoto, R.S. (2011). Mangrove bacterial diversity and the impact of oil contamination revealed by pyrosequencing: bacterial proxies for oil pollution. PloS one, 6(3), e16943. https://doi.org/10.1371/journal.pone.0016943
    6. Ebrahimi, M., Falah, M., & Sarikhani, M.R., (2013). Isolation and identification of some bacteria that decompose petroleum substances from soil contaminated with petroleum substances and checking their growth ability in the presence of gasoline. Water and Soil Science, 3(1), 109-121. (In Persian with English abstract)
    7. Gianfreda, L., Rao, M.A., Piotrowska, A., Palumbo, G., & Colombo, C. (2005). Soil enzyme activities as affected by anthropogenic alterations: intensive agricultural practices and organic pollution. Science of the Total Environment, 341(1-3), 265-279. https://doi.org/10.1016/j.scitotenv.2004.10.005
    8. Guo, H., Yao, J., Cai, M., Qian, Y., Guo, Y., Richnow, H.H., ... & Ceccanti, B. (2012). Effects of petroleum contamination on soil microbial numbers, metabolic activity and urease activity. Chemosphere, 87(11), 1273-1280. https://doi.org/10.1016/j.chemosphere.2012.01.034
    9. Hui, L.I., Zhang, Y., Kravchenko, I., Hui, X.U., & Zhang, C.G. (2007). Dynamic changes in microbial activity and community structure during biodegradation of petroleum compounds: a laboratory experiment. Journal of Environmental Sciences, 19(8), 1003-1013. https://doi.org/10.1016/S1001-0742(07)60163-6
    10. Klamerus-Iwan, A., Błońska, E., Lasota, J., Kalandyk, A., & Waligórski, P. (2015). Influence of oil contamination on physical and biological properties of forest soil after chainsaw use. Water, Air, & Soil Pollution, 226, 1-9. https://doi.org/10.1007/s11270-015-2649-2
    11. Labud, V., Garcia, C., & Hernandez, T. (2007). Effect of hydrocarbon pollution on the microbial properties of a sandy and a clay soil. Chemosphere, 66(10), 1863-1871. https://doi.org/10.1016/j.chemosphere.2006.08.021
    12. Lee, S.H., Oh, B.I., & Kim, J.G. (2008). Effect of various amendments on heavy mineral oil bioremediation and soil microbial activity. Bioresource Technology, 99(7), 2578-2587. https://doi.org/10.1016/j.biortech.2007.04.039
    13. Liang, Y., Zhang, X., Zhou, J., & Li, G. (2015). Long‐term oil contamination increases deterministic assembly processes in soil microbes. Ecological Applications, 25(5), 1235-1243. https://doi.org/10.1890/14-1672.1
    14. Liao, J., Wang, J., Jiang, D., Wang, M.C., & Huang, Y. (2015). Long-term oil contamination causes similar changes in microbial communities of two distinct soils. Applied Microbiology and Biotechnology, 99, 10299-10310. https://doi.org/10.1007/s00253-015-6880-y
    15. Lipińska, A., Kucharski, J., & Wyszkowska, J. (2013). Urease activity in soil contaminated with polycyclic aromatic hydrocarbons. Polish Journal of Environmental Studies, 22(5), 1393-1400.
    16. Margesin, R., Hämmerle, M., & Tscherko, D. (2007). Microbial activity and community composition during bioremediation of diesel-oil-contaminated soil: effects of hydrocarbon concentration, fertilizers, and incubation time. Microbial Ecology, 53, 259-269. https://doi.org/10.1007/s00248-006-9136-7
    17. Martin, A.E., & Reeve, R. (1955). A rapid manometeic method for determining soil carbonate. Soil Science, 79(3), 187-198.
    18. Moradi, S. H., Sarikhani, M. R., & Alliasgharzad, N. (2019). Isolation of endophytic bacteria from grasses root and assessing phosphate solubilization, potassium releasing and auxin production abilities of isolated bacteria. Biological Journal of Microorganism, 36(2020), 1-13. https://doi.org/10.22108/bjm.2019.117674.1207
    19. Moreno, B., Nogales, R., Macci, C., Masciandaro, G., & Benitez, E. (2011). Microbial eco-physiological profiles to estimate the biological restoration of a trichloroethylene-contaminated soil. Ecological Indicators 11(6), 1563-1571. https://doi.org/10.1016/j.ecolind.2011.03.026
    20. Nie, M., Zhang, X.D., Wang, J.Q., Jiang, L.F., Yang, J., Quan, Z.X., ... & Li, B. (2009). Rhizosphere effects on soil bacterial abundance and diversity in the Yellow River Deltaic ecosystem as influenced by petroleum contamination and soil salinization. Soil Biology and Biochemistry, 41(12), 2535-2542. https://doi.org/10.1016/ j.soilbio.2009.09.012
    21. Phillips, L.A., Greer, C.W., Farrell, R.E., & Germida, J.J. (2009). Field-scale assessment of weathered hydrocarbon degradation by mixed and single plant treatments. Applied Soil Ecology, 42(1), 9-17. https://doi.org/10.1016/ j.apsoil.2009.01.002
    22. Rao, M.A., Scelza, R., Acevedo, F., Diez, M. C., & Gianfreda, L. (2014). Enzymes as useful tools for environmental purposes. Chemosphere 107: 145-162. https://doi.org/10.1016/j.chemosphere.2013.12.059.
    23. Rowell, D.L. (1994). Soil Science: Methods and Applications. Longman, UK.
    24. Saadoun, I., Mohammad, M.J., Hameed, K.M., & Shawaqfah, M.A. (2008). Microbial populations of crude oil spill polluted soils at the Jordan-Iraq desert (the Badia region). Brazilian Journal of Microbiology, 39, 453-456.
    25. Schinner, F., Öhlinger, R., Kandeler, E., & Margesin, R. (Eds.). (2012). Methods in soil biology. Springer Science & Business Media.
    26. Sutton, N.B., Maphosa, F., Morillo, J.A., Abu Al-Soud, W., Langenhoff, A.A., Grotenhuis, T., Rijnaarts, H.H., & Smidt, H. (2013). Impact of long-term diesel contamination on soil microbial community structure. Applied and Environmental Microbiology, 79(2): 619–630. https://doi.org/10.1128/AEM.02747-12
    27. Tabatabai, M.A. (1994). Soil enzymes. Methods of soil analysis: Part 2 Microbiological and biochemical properties, 5, 775-833. https://doi.org/10.2136/sssabookser5.2.c37
    28. Trasar-Cepeda, C., Leiros, M.C., Seoane, S., & Gil-Sotres, F. (2000). Limitations of soil enzymes as indicators of soil pollution. Soil Biology and Biochemistry, 32(13), 1867-1875. https://doi.org/10.1016/S0038-0717(00)00160-7
    29. Vincent, A.O., Felix, E., Weltime, M.O., Ize-iyamu, O.K., & Daniel, E.E. (2011). Microbial degradation and its kinetics on crude oil polluted soil. Research Journal of Chemical Sciences.
    30. Wyszkowska, J., Kucharski, M., & Kucharski, J. (2006). Application of the activity of soil enzymes in the evaluation of soil contamination by Diesel oil. Polish Journal of Environmental Studies, 15(3), 501-506.
    31. Xiao, K.Q., Li, L.G., Ma, L.P., Zhang, S.Y., Bao, P., Zhang, T., & Zhu, Y.G. (2016). Metagenomic analysis revealed highly diverse microbial arsenic metabolism genes in paddy soils with low-arsenic contents. Environmental Pollution, 211, 1-8. https://doi.org/10.1016/j.envpol.2015.12.023
    32. Xu, J.G., & Johnson, R.L. (1995). Root growth, microbial activity and phosphatase activity in oil-contaminated, remediated and uncontaminated soils planted to barley and field pea. Plant and Soil, 173(1), 3-10. https://doi.org/10.1007/BF00155512
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