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Bioremediation of petroleum contaminated environments is a process in which the biological pathways within microorganisms or plants are used to degrade or sequester toxic hydrocarbons, heavy metals, and other volatile organic compounds found within fossil fuels. Oil spills happen frequently at varying degrees along with all aspects of the petroleum supply chain, presenting a complex array of issues for both environmental and public health.<ref>{{Cite journal|last=McGill|first=W.B.|date=1977-04-01|title=Soil Restoration Following Oil Spills - A Review|url=http://dx.doi.org/10.2118/77-02-07|journal=Journal of Canadian Petroleum Technology|volume=16|issue=02|doi=10.2118/77-02-07|issn=0021-9487}}</ref> While traditional cleanup methods such as chemical or manual containment and removal often result in rapid results, bioremediation is less labor-intensive, expensive, and averts chemical or mechanical damage.<ref>{{Cite journal|last=Dave|date=2011-05-01|title=Remediation Technologies for Marine Oil Spills: A Critical Review and Comparative Analysis|url=http://dx.doi.org/10.3844/ajessp.2011.423.440|journal=American Journal of Environmental Sciences|volume=7|issue=5|pages=423–440|doi=10.3844/ajessp.2011.423.440|issn=1553-345X}}</ref><ref>{{Cite journal|last=Walls|first=W.D.|date=2010-05|title=Petroleum refining industry in China|url=http://dx.doi.org/10.1016/j.enpol.2009.06.002|journal=Energy Policy|volume=38|issue=5|pages=2110–2115|doi=10.1016/j.enpol.2009.06.002|issn=0301-4215}}</ref><ref>{{Cite journal|last=YANG|first=Si-Zhong|last2=JIN|first2=Hui-Jun|last3=WEI|first3=Zhi|last4=HE|first4=Rui-Xia|last5=JI|first5=Yan-Jun|last6=LI|first6=Xiu-Mei|last7=YU|first7=Shao-Peng|date=2009-06|title=Bioremediation of Oil Spills in Cold Environments: A Review|url=http://dx.doi.org/10.1016/s1002-0160(09)60128-4|journal=Pedosphere|volume=19|issue=3|pages=371–381|doi=10.1016/s1002-0160(09)60128-4|issn=1002-0160}}</ref> The efficiency and effectiveness of bioremediation efforts are based on maintaining ideal conditions, such as pH, RED-OX potential, temperature, moisture, oxygen abundance, nutrient availability, soil composition, and pollutant structure, for the desired organism or biological pathway to facilitate reactions.<ref>( Council, National Research (1969-12-31). In Situ Bioremediation: When Does it Work?. [[Doi (identifier)|doi]]:10.17226/2131.[[ISBN (identifier)|ISBN]] [[Special:BookSources/9780309048965|9780309048965]].</ref> Three main types of bioremediation used for petroleum spills include microbial remediation, phytoremediation, and mycoremediation. Bioremediation has been implemented in various notable oil spills including the 1989 Exxon Valdez incident where the application of fertilizer on affected shoreline increased rates of biodegradation.<ref>{{Cite journal|last=Atlas|first=Ronald M.|last2=Hazen|first2=Terry C.|date=2011-08-15|title=Oil Biodegradation and Bioremediation: A Tale of the Two Worst Spills in U.S. History|url=https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3155281/|journal=Environmental Science & Technology|volume=45|issue=16|pages=6709–6715|doi=10.1021/es2013227|issn=0013-936X|pmc=3155281|pmid=21699212}}</ref>


== Introduction to oil spills ==
== Introduction to oil spills ==
Petroleum contamination of both terrestrial and marine environments results from prospection, extraction, refinement, transport, and storage of oil. Oil spills have been a global issue since the emergence of the oil industry in the early 1900s. The risk of unintentional and intentional spillage has increased as the energy industry and global demand expand.<ref>{{Cite journal|last=Sabir|first=Syed|date=2015-09-02|title=Approach of Cost-Effective Adsorbents for Oil Removal from Oily Water|url=http://www.tandfonline.com/doi/full/10.1080/10643389.2014.1001143|journal=Critical Reviews in Environmental Science and Technology|language=en|volume=45|issue=17|pages=1916–1945|doi=10.1080/10643389.2014.1001143|issn=1064-3389}}</ref> Petroleum is a toxic mixture of organic compounds, trace amounts of heavy metals, and hydrocarbons including many persistent volatile organic compounds (VOCs) and polycyclic aromatic hydrocarbons (PAHs).<ref>{{Cite book|last=Hooper|first=Craig H.|url=http://dx.doi.org/10.5962/bhl.title.62199|title=The IXTOC I oil spill : the Federal scientific response /|date=1982|publisher=U.S. Dept. of Commerce, National Oceanic and Atmospheric Administration, Office of Marine Pollution Assessment,|location=Boulder, Colo. :}}</ref><ref>IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS) e-ISSN:2278-3008, p-ISSN:2319-7676. Volume 11, Issue 3 Ver. IV (May - Jun.2016), PP 43-44 www.iosrjournals.org DOI: 10.9790/3008-1103044344 www.iosrjournals.org 43 | Page Profile of Heavy Metals in Crude Oil Commonly Consumed For Medicinal Purposes in Abakaliki OTI Wilberforce J.O Department of Industrial Chemistry, Ebonyi State University, P.M.B. 053, Abakaliki, Ebonyi State, Nigeria.</ref>  Discharged into marine environments oil is particularly damaging due to rapid dispersal and the creation of secondary pollutants through photolysis.<ref>{{Cite journal|last=Kingston|first=Paul F|date=2002-06|title=Long-term Environmental Impact of Oil Spills|url=http://dx.doi.org/10.1016/s1353-2561(02)00051-8|journal=Spill Science & Technology Bulletin|volume=7|issue=1-2|pages=53–61|doi=10.1016/s1353-2561(02)00051-8|issn=1353-2561}}</ref> Petroleum bioaccumulation in terrestrial and marine food chains cause both acute and long term health effects. Exposure to oil damages critical functions within organisms including reproduction, regulation of physiological and chemical processes, and organ function.<ref>{{Cite journal|last=Tabari|first=Khashayar|last2=Tabari|first2=Mahsa|date=2010-11|title=Biodegradation of heavy crude oil: effects and some innovative clean-up biotechnologies|url=http://dx.doi.org/10.1016/j.jbiotec.2010.09.220|journal=Journal of Biotechnology|volume=150|pages=285–285|doi=10.1016/j.jbiotec.2010.09.220|issn=0168-1656}}</ref> Large spills alter ecosystem dynamics leading to algae blooms and a mass die-off of marine life.<ref>{{Cite journal|last=Jernelöv|first=Arne|date=2010-7|title=The Threats from Oil Spills: Now, Then, and in the Future|url=https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3357709/|journal=Ambio|volume=39|issue=5-6|pages=353–366|doi=10.1007/s13280-010-0085-5|issn=0044-7447|pmc=3357709|pmid=21053719}}</ref> It is estimated that over 1000 sea otters, along with many birds, died from the Exxon Valdez spill.<ref>{{Cite journal|last=Jernelöv|first=Arne|date=2010-07|title=The Threats from Oil Spills: Now, Then, and in the Future|url=http://link.springer.com/10.1007/s13280-010-0085-5|journal=AMBIO|language=en|volume=39|issue=5-6|pages=353–366|doi=10.1007/s13280-010-0085-5|issn=0044-7447|pmc=PMC3357709|pmid=21053719}}</ref>
The spilling of petroleum into our natural waterways is a major issue, and petroleum causes both acute and long term issues. Oil is harvested and processed in so many ways, there is a lot of room for error. These spills can be caused by tankers, refineries, drilling operations, or even storage facilities.<ref name=":0">{{Cite web|url=http://response.restoration.noaa.gov/oil-and-chemical-spills|title=Oil and Chemical Spills|last=|first=|date=|website=response.restoration.noaa.gov|language=en|archive-url=|archive-date=|access-date=2017-02-19}}</ref> Regardless of how petroleum is released into nature, spills account for high percentages of marine death and complicate vegetative life.<ref name=":0" /> When oil leaks into an ecosystem, it alters the balance of both the habitat and the organisms that live there. These organisms experience altered growth and reproduction patterns, anatomical complications, and increased susceptibility to hypothermia.<ref>{{Cite web|url=http://oceanservice.noaa.gov/facts/oilimpacts.html|title=How does oil impact marine life?|last=Administration|first=US Department of Commerce, National Oceanic and Atmospheric|website=oceanservice.noaa.gov|language=EN-US|access-date=2017-02-19}}</ref> The water resistant properties of birds and mammals are compromised due to matting of fur and exposed skin.<ref>{{Cite web|url=https://www.bird-rescue.org/our-work/research-and-education/how-oil-affects-birds.aspx|title=How Oil Affects Birds|last=|first=|date=2011|website=IBR|archive-url=|archive-date=}}</ref>

Along with this, petroleum is toxic. Petroleum, or crude oil, is a complex mixture of many hydrocarbons and can be used as raw material or lubricant.<ref>{{Cite news|url=https://www.britannica.com/science/petroleum|title=petroleum|newspaper=Encyclopedia Britannica|access-date=2017-02-19|language=en}}</ref> It is known to contain upwards of 17,000 organic compounds<ref name=":6">{{Cite journal|last=Brooijmans|first=Rob J. W.|last2=Pastink|first2=Margreet I.|last3=Siezen|first3=Roland J.|date=2009-11-01|title=Hydrocarbon-degrading bacteria: the oil-spill clean-up crew|journal=Microbial Biotechnology|language=en|volume=2|issue=6|pages=587–594|doi=10.1111/j.1751-7915.2009.00151.x|issn=1751-7915|pmc=3815313|pmid=21255292}}</ref> and both [[volatile organic compounds]] (VOC's), and [[polycyclic aromatic hydrocarbon]]s (PAH's).<ref name=":1">{{Cite web|url=http://response.restoration.noaa.gov/about/media/toxicity-oil-whats-big-deal.html|title=The Toxicity of Oil: What's the Big Deal? {{!}} response.restoration.noaa.gov|website=response.restoration.noaa.gov|language=en|access-date=2017-02-19}}</ref> VOC's can be carcinogenic and evaporate into the air very easily, making them toxic when taken in.<ref name=":1" /> PAH's have some of the same effects, and can last much longer in the environment.<ref name=":1" /> In many expansive oil spills, such as the ''[[Exxon Valdez oil spill|Exxon Valdez]]'' or ''[[Deepwater Horizon oil spill|BP Deepwater Horizon]]'' incidents, substantial effects were seen when petroleum was released into those marine ecosystems.<ref name=":3">{{Cite journal|last=Atlas|first=Ronald M.|last2=Hazen|first2=Terry C.|date=2011-08-15|title=Oil Biodegradation and Bioremediation: A Tale of the Two Worst Spills in U.S. History|journal=Environmental Science & Technology|volume=45|issue=16|pages=6709–6715|doi=10.1021/es2013227|issn=0013-936X|pmc=3155281|pmid=21699212}}</ref>{{TOC right }}


== Bioremediation ==
== Bioremediation ==
Bioremediation refers to the use of specific microorganisms to metabolize and remove harmful substances. These microorganisms are known for their affinity for hydrocarbons, both biochemically and physically. They have the ability to effectively recycle a wide range of compounds and hydrocarbons by converting them into non-toxic components.<ref>https://bionextechnology.com/</ref> In most cases, bioremediation works to either increase the numbers of naturally occurring microorganisms or add pollutant-specific microbes to the area.<ref name=":2">{{Cite web|url=http://ei.cornell.edu/biodeg/bioremed/|title=Environmental Inquiry - Bioremediation|website=ei.cornell.edu|access-date=2017-02-19}}</ref> [[Bioremediation]] can involve using many varieties of [[microorganism]]s as well, either synergistically or independently of each other. Either way, bioremediation is commonly better for the environment and less expensive than other chemical methods.<ref name=":2" /> Different types of bacteria, archaea, algae, and some species of plants are all able to breakdown specific toxic waste products into safer constituents. However, specific bacteria mostly commonly are used in bioremediation of oil. These specific microorganisms are not hard to find, either. There are many types of bacteria and archaea that evolve to degrade all types of substances, including petroleum.<ref name=":3" />
Bioremediation refers to the use of specific microorganisms or plants to metabolize and remove harmful substances. These organisms are known for their biochemical and physical affinity to hydrocarbons among other pollutants. Various types of bacteria, archaea, algae, fungi, and some species of plants are all able to break down specific toxic waste products into safer constituents. Bioremediation is classified by the organism responsible for remediation with three major subdivisions: microbial remediation, phytoremediation, and mycoremediation.<ref>{{Citation|last=Shukla|first=Anurakti|title=Emerging Aspects of Bioremediation of Arsenic|date=2017|url=http://dx.doi.org/10.1007/978-3-319-50654-8_17|work=Green Technologies and Environmental Sustainability|pages=395–407|publisher=Springer International Publishing|isbn=978-3-319-50653-1|access-date=2020-06-01|last2=Srivastava|first2=Sudhakar}}</ref> In most cases, bioremediation works to either increase the numbers of naturally occurring microorganisms or add pollutant-specific microbes to the area. [[Bioremediation]] can involve using many varieties of [[Microorganism|microorganisms]] as well, either synergistically or independently of each other. The costs and environmental impacts of bioremediation are often negligible when compared to traditional manual or chemical remediation efforts.<ref>{{Cite journal|last=Borasiya|first=Hiral|last2=Shah|first2=Maulin P|date=2017|title=Waste Water Treatment by Environmental Microbiology|url=http://dx.doi.org/10.4172/2155-6199.1000386|journal=Journal of Bioremediation & Biodegradation|volume=08|issue=02|doi=10.4172/2155-6199.1000386|issn=2155-6199}}</ref>


== Bioremediation of petroleum ==
== Bioremediation of petroleum ==
Due to their ubiquity across environments, many organisms have evolved to use the hydrocarbons and organic compounds in petroleum as energy while simultaneously denaturing toxins through molecular transfer mechanisms.<ref>{{Cite book|url=http://www.nap.edu/catalog/2131|title=In Situ Bioremediation: When Does it Work?|date=1993-01-01|publisher=National Academies Press|isbn=978-0-309-04896-5|location=Washington, D.C.|doi=10.17226/2131. isbn 9780309048965.)}}</ref>
Oil degrading organisms have evolved to use the hydrocarbons and organic compounds in petroleum as energy, and use molecular transfer mechanisms to denature these toxins.<ref name=":4">{{Cite book|title=In Situ Bioremediation: When Does it Work?|last=Council|first=National Research|date=1969-12-31|isbn=9780309048965|language=en|doi=10.17226/2131}}</ref> The aerobic and anaerobic properties of these microbes allow them to respire and ferment compounds as well, and this tends to result in the transformation of toxins into innocuous compounds.<ref name=":4" /> These compounds have more stable pH levels, increased solubility in water, and are less aggressive molecularly. It is known that the composition of oil-degrading microorganisms in marine ecosystems is originally less than 1%. When these organisms are given the necessary substrate, they tend to thrive and grow to almost 10% of the complete microbiome.<ref name=":6" /> Dependent on physical and chemical properties, petroleum-degenerative microorganisms take longer to degrade high-molecular weighted compounds, such as polycyclic aromatic hydrocarbons (PAH's). These microbes require a wide array of enzymes for the breakdown of petroleum, and require very specific nutrient composition to work at an efficient rate.<ref name=":7">{{Cite journal|last=Das|first=Nilanjana|last2=Chandran|first2=Preethy|date=2010-09-13|title=Microbial Degradation of Petroleum Hydrocarbon Contaminants: An Overview|journal=Biotechnology Research International|language=en|volume=2011|pages=941810|doi=10.4061/2011/941810|issn=2090-3138|pmc=3042690|pmid=21350672}}</ref>


Microbial bioremediation uses aerobic and anaerobic properties of various microbes to respire and ferment compounds transforming toxins into innocuous compounds.<ref>{{Cite book|url=http://www.nap.edu/catalog/2131|title=In Situ Bioremediation: When Does it Work?|date=1993-01-01|publisher=National Academies Press|isbn=978-0-309-04896-5|location=Washington, D.C.|doi=10.17226/2131. isbn 9780309048965.)}}</ref> These resulting compounds exhibit more neutral pH levels, increased solubility in water, and are less reactive molecularly. Baseline populations of oil-degrading microorganisms typically account for less than 1% of microbiomes associated with marine ecosystems. Remediation techniques which remove reaction limiting factors through the addition of substrate, can boots microbe population towards 10% of the ecosystems microbiome.<ref>Brooijmans, Rob J. W.; Pastink, Margreet I.; Siezen, Roland J. (2009-11-01). "Hydrocarbon-degrading bacteria: the oil-spill clean-up crew". Microbial Biotechnology. 2 (6): 587–594. [[Doi (identifier)|doi]]:10.1111/j.1751-7915.2009.00151.x. [[ISSN (identifier)|ISSN]] 1751-7915. [[PMC (identifier)|PMC]] 3815313. [[PMID (identifier)|PMID]] 21255292.</ref> Dependent on physical and chemical properties, petroleum-degenerative microorganisms take longer to degrade compounds with high-molecular-weight, such as polycyclic aromatic hydrocarbons (PAH's). These microbes require a wide array of enzymes for the breakdown of petroleum, and very specific nutrient compositions to work at an efficient rate.<ref name=":0">Das, Nilanjana; Chandran, Preethy (2010-09-13). "Microbial Degradation of Petroleum Hydrocarbon Contaminants: An Overview". Biotechnology Research International. 2011: 941810. [[Doi (identifier)|doi]]:10.4061/2011/941810. [[ISSN (identifier)|ISSN]] 2090-3138. [[PMC (identifier)|PMC]] 3042690. [[PMID (identifier)|PMID]] 21350672.</ref>
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Microbes work in a step-wise fashion to breakdown and metabolize the components of petroleum.<ref name=":7" />

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# Linear Alkanes

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# Branched Alkanes
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Microbes work in a step-wise fashion to breakdown and metabolize the components of petroleum.<ref name=":0" />
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# Small aromatic compounds

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# Linear Alkanes
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# Branched Alkanes
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# Small aromatic compounds
# Cyclic Alkanes
# Cyclic Alkanes


Treatments that use these breakdown processes most commonly use heat and chemicals to extend the efficacy.<ref name=":8">{{Cite journal|last=Al Disi|first=Zulfa|last2=Jaoua|first2=Samir|last3=Al-Thani|first3=Dhabia|last4=Al-Meer|first4=Saeed|last5=Zouari|first5=Nabil|date=2017-01-24|title=Considering the Specific Impact of Harsh Conditions and Oil Weathering on Diversity, Adaptation, and Activity of Hydrocarbon-Degrading Bacteria in Strategies of Bioremediation of Harsh Oily-Polluted Soils|journal=BioMed Research International|language=en|volume=2017|pages=8649350|doi=10.1155/2017/8649350|issn=2314-6133|pmc=5294359|pmid=28243605}}</ref> Later, more biological systems are used for specific ecosystems that use specific mechanisms.<ref name=":8" />
Treatments that use these breakdown processes most commonly use heat and chemicals to extend the efficacy.<ref name=":1">Al Disi, Zulfa; Jaoua, Samir; Al-Thani, Dhabia; Al-Meer, Saeed; Zouari, Nabil (2017-01-24). "Considering the Specific Impact of Harsh Conditions and Oil Weathering on Diversity, Adaptation, and Activity of Hydrocarbon-Degrading Bacteria in Strategies of Bioremediation of Harsh Oily-Polluted Soils". BioMed Research International. 2017: 8649350. [[Doi (identifier)|doi]]:10.1155/2017/8649350. [[ISSN (identifier)|ISSN]] 2314-6133. [[PMC (identifier)|PMC]] 5294359. [[PMID (identifier)|PMID]] 28243605</ref> Later, more biological systems are used for specific ecosystems that use specific mechanisms.<ref name=":1" />

[[File:Biodegradation_of_Pollutants.png|thumb|362x362px|Mechanisms involved in bioremediation of toxic compounds.]]
Phytoremediation is a process in which plants are used to sequester toxins and hydrocarbons into plant tissue from contaminated soils. The main mechanisms for phytoremediation stem from complex relationships between roots and rhizobia. Plants secrete sugars, enzymes, and oxygen from roots which provide necessary substrates for rhizobia and associated rhizosphere microbes to stimulate degradation of organic pollutants.<ref>{{Cite journal|last=Gerhardt|first=Karen E.|last2=Huang|first2=Xiao-Dong|last3=Glick|first3=Bernard R.|last4=Greenberg|first4=Bruce M.|date=2009-01|title=Phytoremediation and rhizoremediation of organic soil contaminants: Potential and challenges|url=http://dx.doi.org/10.1016/j.plantsci.2008.09.014|journal=Plant Science|volume=176|issue=1|pages=20–30|doi=10.1016/j.plantsci.2008.09.014|issn=0168-9452}}</ref> Studies have demonstrated the bioaccumulation abilities of various plants with rhizobial associations, in particular ''Chromolaena odorata'' were able to remove 80% of petroleum and heavy metal toxins from soils.<ref>{{Cite journal|last=Atagana|first=Harrison Ifeanyichukwu|date=2010-06-05|title=Bioremediation of Co-contamination of Crude Oil and Heavy Metals in Soil by Phytoremediation Using Chromolaena odorata (L) King & H.E. Robinson|url=http://dx.doi.org/10.1007/s11270-010-0476-z|journal=Water, Air, & Soil Pollution|volume=215|issue=1-4|pages=261–271|doi=10.1007/s11270-010-0476-z|issn=0049-6979}}</ref> While more commonly used on terrestrial environments, contaminated marine environments also benefit from plants based bioremediation through the use of various algae and macrophytes.<ref>{{Cite journal|last=Ndimele|first=P.E.|last2=Kumolu-Joh|first2=C.A.|last3=Anetekhai|first3=M.A.|date=2011-06-01|title=The Invasive Aquatic Macrophyte, Water Hyacinth {Eichhornia crassipes (Mart.) Solm-Laubach: Pontedericeae}: Problems and Prospects|url=http://dx.doi.org/10.3923/rjes.2011.509.520|journal=Research Journal of Environmental Sciences|volume=5|issue=6|pages=509–520|doi=10.3923/rjes.2011.509.520|issn=1819-3412}}</ref> Phytoremediation is most effective when used in conjunction with microbial remediation and Mycoremediation.<ref>{{Cite journal|last=Vouillamoz|first=J.|last2=Milke|first2=M. W.|date=2001-01-01|title=Effect of compost in phytoremediation of diesel-contaminated soils|url=http://dx.doi.org/10.2166/wst.2001.0102|journal=Water Science and Technology|volume=43|issue=2|pages=291–295|doi=10.2166/wst.2001.0102|issn=0273-1223}}</ref><ref>{{Cite journal|last=Alarcón|first=Alejandro|last2=Davies|first2=Fred T.|last3=Autenrieth|first3=Robin L.|last4=Zuberer|first4=David A.|date=2008-07-08|title=Arbuscular Mycorrhiza and Petroleum-Degrading Microorganisms Enhance Phytoremediation of Petroleum-Contaminated Soil|url=http://dx.doi.org/10.1080/15226510802096002|journal=International Journal of Phytoremediation|volume=10|issue=4|pages=251–263|doi=10.1080/15226510802096002|issn=1522-6514}}</ref>

Mycoremediation techniques make use of pollutant tolerant fungi which sequester or denature environmental toxins particularly heavy metals. Toxins are sequestered into highly absorbent molecules such chitin and glucan which are found in fungal cell walls.<ref>{{Citation|last=Shukla|first=Anurakti|title=Emerging Aspects of Bioremediation of Arsenic|date=2017|url=http://dx.doi.org/10.1007/978-3-319-50654-8_17|work=Green Technologies and Environmental Sustainability|pages=395–407|publisher=Springer International Publishing|isbn=978-3-319-50653-1|access-date=2020-06-01|last2=Srivastava|first2=Sudhakar}}</ref> ''Saccharomyces cerevisiae'' (baker's yeast) can be used to remediate heavy metal contaminated marine ecosystems, with 80% to 90% success in the case of arsenic.<ref>{{Cite journal|last=Singh|first=Asha Lata|last2=Sarma|first2=P. N.|date=2010-05-13|title=Removal of Arsenic(III) from Waste Water UsingLactobacillus acidophilus|url=http://dx.doi.org/10.1080/10889861003767050|journal=Bioremediation Journal|volume=14|issue=2|pages=92–97|doi=10.1080/10889861003767050|issn=1088-9868}}</ref> Polycyclic aromatic hydrocarbons (PAH) concentrations of soil samples taken from contaminated oil drilling cuttings in Nigeria have been decreased by 7% to 19% using white rot fungi under experimental conditions.<ref>{{Cite journal|last=Araka|first=Perez P.|last2=Okparanma|first2=Reuben N.|last3=Ayotamuno|first3=Josiah M.|date=2019-10|title=Diagnostic screening of organic contaminant level in solidified/stabilized pre-treated oil-based drill cuttings|url=http://dx.doi.org/10.1016/j.heliyon.2019.e02644|journal=Heliyon|volume=5|issue=10|pages=e02644|doi=10.1016/j.heliyon.2019.e02644|issn=2405-8440}}</ref> Soil contaminated with crude oil displays toxic levels of various heavy metals such as lead, zinc and magnesium. Application of mycoremediation techniques to crude contaminated soils have shown significant reductions of heavy metal concentrations.<ref>{{Cite journal|last=Maduekwe|first=C.|last2=Nwachukwu|first2=E. O.|last3=Joel|first3=O. F.|date=2016|title=Comparative Study of Rena and Mycoremediation Techniques in Reduction of Heavy Metals in Crude Oil Impacted Soil|url=http://dx.doi.org/10.2118/184348-ms|journal=SPE Nigeria Annual International Conference and Exhibition|publisher=Society of Petroleum Engineers|doi=10.2118/184348-ms}}</ref>[[File:Biodegradation_of_Pollutants.png|thumb|362x362px|Mechanisms involved in bioremediation of toxic compounds.]]


=== Bioremediation parameters ===
=== Bioremediation parameters ===
The efficiency and efficacy of each method of remediation has limitations. The goal of remediation is to eliminate the environmental pollutant as quickly as possible; only inefficient processes require human intervention.<ref name=":8" /> Environmental factors such as requirements of reaction, mobility of substances, and physiological needs of organisms will affect the rate and degree that contaminants are degraded.<ref name=":4" /> Over time, many of these requirements are overcome. This is when petroleum degrading bacteria and archaea are able to mediate oil spills most efficiently. Weathering and environmental factors play large roles in the success of bioremediation. Interacting soil and pollutant chemicals truly account for the work that can be completed by these microorganisms. These processes change the soil composition and layering, along with the biochemistry of the ecosystem. These chemical and biological changes require adaptation from soil microbes to bioremediate.<ref name=":8" /> The susceptibility of the pollutant is also important to consider. Properties such as solubility, temperature, and pH will affect bioremediation and affect the process.<ref name=":5">{{Cite journal|last=Gkorezis|first=Panagiotis|last2=Daghio|first2=Matteo|last3=Franzetti|first3=Andrea|last4=Hamme|first4=Van|last5=D|first5=Jonathan|last6=Sillen|first6=Wouter|last7=Vangronsveld|first7=Jaco|date=2016-01-01|title=The Interaction between Plants and Bacteria in the Remediation of Petroleum Hydrocarbons: An Environmental Perspective|journal=Frontiers in Microbiology|language=English|volume=7|pages=1836|doi=10.3389/fmicb.2016.01836|issn=1664-302X|pmc=5116465|pmid=27917161}}</ref> Pollutants that are more soluble will be easier for microbes to transform into the environment. Otherwise, pollutants with rigid molecular structures extend bioremediation as they are harder to convert into innocuous substances. Bioaccesability, the amount of pollutant available for absorption, and [[bioavailability]] of pollutant will affect efficiency as well.<ref name=":5" /> In many instances, needed nutrients are collected and allocated for petroleum degrading microorganisms in order to maximize the efficiency of the process.<ref name=":8" /> Providing microorganisms with the nutrients and conditions they need allow them to thrive.
The efficiency and efficacy of each method of remediation has limitations. The goal of remediation is to eliminate the environmental pollutant as quickly as possible; only inefficient processes require human intervention.<ref name=":8">{{Cite journal|last=Al Disi|first=Zulfa|last2=Jaoua|first2=Samir|last3=Al-Thani|first3=Dhabia|last4=Al-Meer|first4=Saeed|last5=Zouari|first5=Nabil|date=2017-01-24|title=Considering the Specific Impact of Harsh Conditions and Oil Weathering on Diversity, Adaptation, and Activity of Hydrocarbon-Degrading Bacteria in Strategies of Bioremediation of Harsh Oily-Polluted Soils|journal=BioMed Research International|language=en|volume=2017|pages=8649350|doi=10.1155/2017/8649350|issn=2314-6133|pmc=5294359|pmid=28243605}}</ref> Environmental factors such as requirements of reaction, mobility of substances, and physiological needs of organisms will affect the rate and degree that contaminants are degraded.<ref name=":4">{{Cite book|last=Council|first=National Research|title=In Situ Bioremediation: When Does it Work?|date=1969-12-31|isbn=9780309048965|language=en|doi=10.17226/2131}}</ref> Over time, many of these requirements are overcome. This is when petroleum degrading bacteria and archaea are able to mediate oil spills most efficiently. Weathering and environmental factors play large roles in the success of bioremediation. Interacting soil and pollutant chemicals truly account for the work that can be completed by these microorganisms. These processes change the soil composition and layering, along with the biochemistry of the ecosystem. These chemical and biological changes require adaptation from soil microbes to bioremediate.<ref name=":8" /> The susceptibility of the pollutant is also important to consider. Properties such as solubility, temperature, and pH will affect bioremediation and affect the process.<ref name=":5">{{Cite journal|last=Gkorezis|first=Panagiotis|last2=Daghio|first2=Matteo|last3=Franzetti|first3=Andrea|last4=Hamme|first4=Van|last5=D|first5=Jonathan|last6=Sillen|first6=Wouter|last7=Vangronsveld|first7=Jaco|date=2016-01-01|title=The Interaction between Plants and Bacteria in the Remediation of Petroleum Hydrocarbons: An Environmental Perspective|journal=Frontiers in Microbiology|language=English|volume=7|pages=1836|doi=10.3389/fmicb.2016.01836|issn=1664-302X|pmc=5116465|pmid=27917161}}</ref> Pollutants that are more soluble will be easier for microbes to transform into the environment. Otherwise, pollutants with rigid molecular structures extend bioremediation as they are harder to convert into innocuous substances. Bioaccessibility, the amount of pollutant available for absorption, and [[bioavailability]] of pollutant will affect efficiency as well.<ref name=":5" /> In many instances, needed nutrients are collected and allocated for petroleum degrading microorganisms in order to maximize the efficiency of the process.<ref name=":8" /> Providing microorganisms with the nutrients and conditions they need allow them to thrive.


=== Factors that affect bioremediation<ref name=":8" /> ===
=== Factors that affect bioremediation<ref name=":8" /> ===

Revision as of 09:25, 1 June 2020

Bioremediation of petroleum contaminated environments is a process in which the biological pathways within microorganisms or plants are used to degrade or sequester toxic hydrocarbons, heavy metals, and other volatile organic compounds found within fossil fuels. Oil spills happen frequently at varying degrees along with all aspects of the petroleum supply chain, presenting a complex array of issues for both environmental and public health.[1] While traditional cleanup methods such as chemical or manual containment and removal often result in rapid results, bioremediation is less labor-intensive, expensive, and averts chemical or mechanical damage.[2][3][4] The efficiency and effectiveness of bioremediation efforts are based on maintaining ideal conditions, such as pH, RED-OX potential, temperature, moisture, oxygen abundance, nutrient availability, soil composition, and pollutant structure, for the desired organism or biological pathway to facilitate reactions.[5] Three main types of bioremediation used for petroleum spills include microbial remediation, phytoremediation, and mycoremediation. Bioremediation has been implemented in various notable oil spills including the 1989 Exxon Valdez incident where the application of fertilizer on affected shoreline increased rates of biodegradation.[6]

Introduction to oil spills

Petroleum contamination of both terrestrial and marine environments results from prospection, extraction, refinement, transport, and storage of oil. Oil spills have been a global issue since the emergence of the oil industry in the early 1900s. The risk of unintentional and intentional spillage has increased as the energy industry and global demand expand.[7] Petroleum is a toxic mixture of organic compounds, trace amounts of heavy metals, and hydrocarbons including many persistent volatile organic compounds (VOCs) and polycyclic aromatic hydrocarbons (PAHs).[8][9]  Discharged into marine environments oil is particularly damaging due to rapid dispersal and the creation of secondary pollutants through photolysis.[10] Petroleum bioaccumulation in terrestrial and marine food chains cause both acute and long term health effects. Exposure to oil damages critical functions within organisms including reproduction, regulation of physiological and chemical processes, and organ function.[11] Large spills alter ecosystem dynamics leading to algae blooms and a mass die-off of marine life.[12] It is estimated that over 1000 sea otters, along with many birds, died from the Exxon Valdez spill.[13]

Bioremediation

Bioremediation refers to the use of specific microorganisms or plants to metabolize and remove harmful substances. These organisms are known for their biochemical and physical affinity to hydrocarbons among other pollutants. Various types of bacteria, archaea, algae, fungi, and some species of plants are all able to break down specific toxic waste products into safer constituents. Bioremediation is classified by the organism responsible for remediation with three major subdivisions: microbial remediation, phytoremediation, and mycoremediation.[14] In most cases, bioremediation works to either increase the numbers of naturally occurring microorganisms or add pollutant-specific microbes to the area. Bioremediation can involve using many varieties of microorganisms as well, either synergistically or independently of each other. The costs and environmental impacts of bioremediation are often negligible when compared to traditional manual or chemical remediation efforts.[15]

Bioremediation of petroleum

Due to their ubiquity across environments, many organisms have evolved to use the hydrocarbons and organic compounds in petroleum as energy while simultaneously denaturing toxins through molecular transfer mechanisms.[16]

Microbial bioremediation uses aerobic and anaerobic properties of various microbes to respire and ferment compounds transforming toxins into innocuous compounds.[17] These resulting compounds exhibit more neutral pH levels, increased solubility in water, and are less reactive molecularly. Baseline populations of oil-degrading microorganisms typically account for less than 1% of microbiomes associated with marine ecosystems. Remediation techniques which remove reaction limiting factors through the addition of substrate, can boots microbe population towards 10% of the ecosystems microbiome.[18] Dependent on physical and chemical properties, petroleum-degenerative microorganisms take longer to degrade compounds with high-molecular-weight, such as polycyclic aromatic hydrocarbons (PAH's). These microbes require a wide array of enzymes for the breakdown of petroleum, and very specific nutrient compositions to work at an efficient rate.[19]


Microbes work in a step-wise fashion to breakdown and metabolize the components of petroleum.[19]

  1. Linear Alkanes
  2. Branched Alkanes
  3. Small aromatic compounds
  4. Cyclic Alkanes

Treatments that use these breakdown processes most commonly use heat and chemicals to extend the efficacy.[20] Later, more biological systems are used for specific ecosystems that use specific mechanisms.[20]

Phytoremediation is a process in which plants are used to sequester toxins and hydrocarbons into plant tissue from contaminated soils. The main mechanisms for phytoremediation stem from complex relationships between roots and rhizobia. Plants secrete sugars, enzymes, and oxygen from roots which provide necessary substrates for rhizobia and associated rhizosphere microbes to stimulate degradation of organic pollutants.[21] Studies have demonstrated the bioaccumulation abilities of various plants with rhizobial associations, in particular Chromolaena odorata were able to remove 80% of petroleum and heavy metal toxins from soils.[22] While more commonly used on terrestrial environments, contaminated marine environments also benefit from plants based bioremediation through the use of various algae and macrophytes.[23] Phytoremediation is most effective when used in conjunction with microbial remediation and Mycoremediation.[24][25]

Mycoremediation techniques make use of pollutant tolerant fungi which sequester or denature environmental toxins particularly heavy metals. Toxins are sequestered into highly absorbent molecules such chitin and glucan which are found in fungal cell walls.[26] Saccharomyces cerevisiae (baker's yeast) can be used to remediate heavy metal contaminated marine ecosystems, with 80% to 90% success in the case of arsenic.[27] Polycyclic aromatic hydrocarbons (PAH) concentrations of soil samples taken from contaminated oil drilling cuttings in Nigeria have been decreased by 7% to 19% using white rot fungi under experimental conditions.[28] Soil contaminated with crude oil displays toxic levels of various heavy metals such as lead, zinc and magnesium. Application of mycoremediation techniques to crude contaminated soils have shown significant reductions of heavy metal concentrations.[29]

Mechanisms involved in bioremediation of toxic compounds.

Bioremediation parameters

The efficiency and efficacy of each method of remediation has limitations. The goal of remediation is to eliminate the environmental pollutant as quickly as possible; only inefficient processes require human intervention.[30] Environmental factors such as requirements of reaction, mobility of substances, and physiological needs of organisms will affect the rate and degree that contaminants are degraded.[31] Over time, many of these requirements are overcome. This is when petroleum degrading bacteria and archaea are able to mediate oil spills most efficiently. Weathering and environmental factors play large roles in the success of bioremediation. Interacting soil and pollutant chemicals truly account for the work that can be completed by these microorganisms. These processes change the soil composition and layering, along with the biochemistry of the ecosystem. These chemical and biological changes require adaptation from soil microbes to bioremediate.[30] The susceptibility of the pollutant is also important to consider. Properties such as solubility, temperature, and pH will affect bioremediation and affect the process.[32] Pollutants that are more soluble will be easier for microbes to transform into the environment. Otherwise, pollutants with rigid molecular structures extend bioremediation as they are harder to convert into innocuous substances. Bioaccessibility, the amount of pollutant available for absorption, and bioavailability of pollutant will affect efficiency as well.[32] In many instances, needed nutrients are collected and allocated for petroleum degrading microorganisms in order to maximize the efficiency of the process.[30] Providing microorganisms with the nutrients and conditions they need allow them to thrive.

Factors that affect bioremediation[30]

  • pH
  • RED-OX reaction potential
  • Temperature
  • Moisture
  • Oxygen and other molecules present
  • Nutrient availability
  • Soil composition
  • Solubility of pollutant

Bioremediation mechanisms

Microorganisms use many unique mechanisms to convert molecules and transfer electrons.[31]
Bioremediation Technique Conversion Products
Aerobic Respiration Petroleum substrate with molecular oxygen Nitrogen Gas, Hydrogen Sulfide,

Methane, Metals, Carbon Dioxide, Water

Inorganic Electron Donation Ammonium, Nitrite, Iron, Manganese are oxidized. Nitrate, Nitrite, Iron, Manganese, Sulfate
Fermentation Toxic petroleum compounds of organic nature Harmless Compounds, Fermentation Products
Demobilization Iron, Sulfate, Mercury, Chromium, Uranium Ferric Hydroxide, Sulfide, Pyrite, Reduced Chromium,

Uraninite

Reductive Dehalogenation Halogen compound with electron donor Reduced contaminant

Listed above, the chemicals required and products formed in petroleum degradation are shown. These microbes will reduce, oxidize, ferment, and demobilize the constituents of oil spills over time, and create innocuous compounds. Bioremediation techniques[33] involve using these mechanisms to reduce pollutant amounts and are dependent on pollutant aspects:

Ex situ bioremediation

Ex situ remediation refers to reactions performed outside the natural habitat of these organisms.

  • Increased microbial activities through aeration, irrigation, and creation of bio-piles.
  • Increased degradation activities via turning of polluted soils and addition of minerals and water.
  • The use of bioreactors, to enhance and speed up the biological reactions of microorganisms to decrease bioremediation time.
  • Farming techniques that call for addition of nutrients in soil to stimulate microbial mechanisms

In situ bioremediation

In situ remediation refers to reactions performed inside a reaction mixture.

  • Bio-venting, using moisture and nutrients to enhance the transformation of pollutants to more innocent substances.
  • Bio-slurping, using pumping to apply oxygen and water, thus separating and compiling soils to increase remediation of microbes.
  • Bio-sparging, where air is pushed into soil to stimulate microbial bioremediation.
  • Phytoremediation, uses the mechanisms of plants to decrease efficacy of pollutants.

References

  1. ^ McGill, W.B. (1977-04-01). "Soil Restoration Following Oil Spills - A Review". Journal of Canadian Petroleum Technology. 16 (02). doi:10.2118/77-02-07. ISSN 0021-9487.
  2. ^ Dave (2011-05-01). "Remediation Technologies for Marine Oil Spills: A Critical Review and Comparative Analysis". American Journal of Environmental Sciences. 7 (5): 423–440. doi:10.3844/ajessp.2011.423.440. ISSN 1553-345X.
  3. ^ Walls, W.D. (2010-05). "Petroleum refining industry in China". Energy Policy. 38 (5): 2110–2115. doi:10.1016/j.enpol.2009.06.002. ISSN 0301-4215. {{cite journal}}: Check date values in: |date= (help)
  4. ^ YANG, Si-Zhong; JIN, Hui-Jun; WEI, Zhi; HE, Rui-Xia; JI, Yan-Jun; LI, Xiu-Mei; YU, Shao-Peng (2009-06). "Bioremediation of Oil Spills in Cold Environments: A Review". Pedosphere. 19 (3): 371–381. doi:10.1016/s1002-0160(09)60128-4. ISSN 1002-0160. {{cite journal}}: Check date values in: |date= (help)
  5. ^ ( Council, National Research (1969-12-31). In Situ Bioremediation: When Does it Work?. doi:10.17226/2131.ISBN 9780309048965.
  6. ^ Atlas, Ronald M.; Hazen, Terry C. (2011-08-15). "Oil Biodegradation and Bioremediation: A Tale of the Two Worst Spills in U.S. History". Environmental Science & Technology. 45 (16): 6709–6715. doi:10.1021/es2013227. ISSN 0013-936X. PMC 3155281. PMID 21699212.
  7. ^ Sabir, Syed (2015-09-02). "Approach of Cost-Effective Adsorbents for Oil Removal from Oily Water". Critical Reviews in Environmental Science and Technology. 45 (17): 1916–1945. doi:10.1080/10643389.2014.1001143. ISSN 1064-3389.
  8. ^ Hooper, Craig H. (1982). The IXTOC I oil spill : the Federal scientific response /. Boulder, Colo. :: U.S. Dept. of Commerce, National Oceanic and Atmospheric Administration, Office of Marine Pollution Assessment,.{{cite book}}: CS1 maint: extra punctuation (link)
  9. ^ IOSR Journal of Pharmacy and Biological Sciences (IOSR-JPBS) e-ISSN:2278-3008, p-ISSN:2319-7676. Volume 11, Issue 3 Ver. IV (May - Jun.2016), PP 43-44 www.iosrjournals.org DOI: 10.9790/3008-1103044344 www.iosrjournals.org 43 | Page Profile of Heavy Metals in Crude Oil Commonly Consumed For Medicinal Purposes in Abakaliki OTI Wilberforce J.O Department of Industrial Chemistry, Ebonyi State University, P.M.B. 053, Abakaliki, Ebonyi State, Nigeria.
  10. ^ Kingston, Paul F (2002-06). "Long-term Environmental Impact of Oil Spills". Spill Science & Technology Bulletin. 7 (1–2): 53–61. doi:10.1016/s1353-2561(02)00051-8. ISSN 1353-2561. {{cite journal}}: Check date values in: |date= (help)
  11. ^ Tabari, Khashayar; Tabari, Mahsa (2010-11). "Biodegradation of heavy crude oil: effects and some innovative clean-up biotechnologies". Journal of Biotechnology. 150: 285–285. doi:10.1016/j.jbiotec.2010.09.220. ISSN 0168-1656. {{cite journal}}: Check date values in: |date= (help)
  12. ^ Jernelöv, Arne (2010-7). "The Threats from Oil Spills: Now, Then, and in the Future". Ambio. 39 (5–6): 353–366. doi:10.1007/s13280-010-0085-5. ISSN 0044-7447. PMC 3357709. PMID 21053719. {{cite journal}}: Check date values in: |date= (help)
  13. ^ Jernelöv, Arne (2010-07). "The Threats from Oil Spills: Now, Then, and in the Future". AMBIO. 39 (5–6): 353–366. doi:10.1007/s13280-010-0085-5. ISSN 0044-7447. PMC 3357709. PMID 21053719. {{cite journal}}: Check date values in: |date= (help)CS1 maint: PMC format (link)
  14. ^ Shukla, Anurakti; Srivastava, Sudhakar (2017), "Emerging Aspects of Bioremediation of Arsenic", Green Technologies and Environmental Sustainability, Springer International Publishing, pp. 395–407, ISBN 978-3-319-50653-1, retrieved 2020-06-01
  15. ^ Borasiya, Hiral; Shah, Maulin P (2017). "Waste Water Treatment by Environmental Microbiology". Journal of Bioremediation & Biodegradation. 08 (02). doi:10.4172/2155-6199.1000386. ISSN 2155-6199.{{cite journal}}: CS1 maint: unflagged free DOI (link)
  16. ^ In Situ Bioremediation: When Does it Work?. Washington, D.C.: National Academies Press. 1993-01-01. doi:10.17226/2131. isbn 9780309048965.). ISBN 978-0-309-04896-5. {{cite book}}: Check |doi= value (help)
  17. ^ In Situ Bioremediation: When Does it Work?. Washington, D.C.: National Academies Press. 1993-01-01. doi:10.17226/2131. isbn 9780309048965.). ISBN 978-0-309-04896-5. {{cite book}}: Check |doi= value (help)
  18. ^ Brooijmans, Rob J. W.; Pastink, Margreet I.; Siezen, Roland J. (2009-11-01). "Hydrocarbon-degrading bacteria: the oil-spill clean-up crew". Microbial Biotechnology. 2 (6): 587–594. doi:10.1111/j.1751-7915.2009.00151.x. ISSN 1751-7915. PMC 3815313. PMID 21255292.
  19. ^ a b Das, Nilanjana; Chandran, Preethy (2010-09-13). "Microbial Degradation of Petroleum Hydrocarbon Contaminants: An Overview". Biotechnology Research International. 2011: 941810. doi:10.4061/2011/941810. ISSN 2090-3138. PMC 3042690. PMID 21350672.
  20. ^ a b Al Disi, Zulfa; Jaoua, Samir; Al-Thani, Dhabia; Al-Meer, Saeed; Zouari, Nabil (2017-01-24). "Considering the Specific Impact of Harsh Conditions and Oil Weathering on Diversity, Adaptation, and Activity of Hydrocarbon-Degrading Bacteria in Strategies of Bioremediation of Harsh Oily-Polluted Soils". BioMed Research International. 2017: 8649350. doi:10.1155/2017/8649350. ISSN 2314-6133. PMC 5294359. PMID 28243605
  21. ^ Gerhardt, Karen E.; Huang, Xiao-Dong; Glick, Bernard R.; Greenberg, Bruce M. (2009-01). "Phytoremediation and rhizoremediation of organic soil contaminants: Potential and challenges". Plant Science. 176 (1): 20–30. doi:10.1016/j.plantsci.2008.09.014. ISSN 0168-9452. {{cite journal}}: Check date values in: |date= (help)
  22. ^ Atagana, Harrison Ifeanyichukwu (2010-06-05). "Bioremediation of Co-contamination of Crude Oil and Heavy Metals in Soil by Phytoremediation Using Chromolaena odorata (L) King & H.E. Robinson". Water, Air, & Soil Pollution. 215 (1–4): 261–271. doi:10.1007/s11270-010-0476-z. ISSN 0049-6979.
  23. ^ Ndimele, P.E.; Kumolu-Joh, C.A.; Anetekhai, M.A. (2011-06-01). "The Invasive Aquatic Macrophyte, Water Hyacinth {Eichhornia crassipes (Mart.) Solm-Laubach: Pontedericeae}: Problems and Prospects". Research Journal of Environmental Sciences. 5 (6): 509–520. doi:10.3923/rjes.2011.509.520. ISSN 1819-3412.
  24. ^ Vouillamoz, J.; Milke, M. W. (2001-01-01). "Effect of compost in phytoremediation of diesel-contaminated soils". Water Science and Technology. 43 (2): 291–295. doi:10.2166/wst.2001.0102. ISSN 0273-1223.
  25. ^ Alarcón, Alejandro; Davies, Fred T.; Autenrieth, Robin L.; Zuberer, David A. (2008-07-08). "Arbuscular Mycorrhiza and Petroleum-Degrading Microorganisms Enhance Phytoremediation of Petroleum-Contaminated Soil". International Journal of Phytoremediation. 10 (4): 251–263. doi:10.1080/15226510802096002. ISSN 1522-6514.
  26. ^ Shukla, Anurakti; Srivastava, Sudhakar (2017), "Emerging Aspects of Bioremediation of Arsenic", Green Technologies and Environmental Sustainability, Springer International Publishing, pp. 395–407, ISBN 978-3-319-50653-1, retrieved 2020-06-01
  27. ^ Singh, Asha Lata; Sarma, P. N. (2010-05-13). "Removal of Arsenic(III) from Waste Water UsingLactobacillus acidophilus". Bioremediation Journal. 14 (2): 92–97. doi:10.1080/10889861003767050. ISSN 1088-9868.
  28. ^ Araka, Perez P.; Okparanma, Reuben N.; Ayotamuno, Josiah M. (2019-10). "Diagnostic screening of organic contaminant level in solidified/stabilized pre-treated oil-based drill cuttings". Heliyon. 5 (10): e02644. doi:10.1016/j.heliyon.2019.e02644. ISSN 2405-8440. {{cite journal}}: Check date values in: |date= (help)
  29. ^ Maduekwe, C.; Nwachukwu, E. O.; Joel, O. F. (2016). "Comparative Study of Rena and Mycoremediation Techniques in Reduction of Heavy Metals in Crude Oil Impacted Soil". SPE Nigeria Annual International Conference and Exhibition. Society of Petroleum Engineers. doi:10.2118/184348-ms.
  30. ^ a b c d Al Disi, Zulfa; Jaoua, Samir; Al-Thani, Dhabia; Al-Meer, Saeed; Zouari, Nabil (2017-01-24). "Considering the Specific Impact of Harsh Conditions and Oil Weathering on Diversity, Adaptation, and Activity of Hydrocarbon-Degrading Bacteria in Strategies of Bioremediation of Harsh Oily-Polluted Soils". BioMed Research International. 2017: 8649350. doi:10.1155/2017/8649350. ISSN 2314-6133. PMC 5294359. PMID 28243605.{{cite journal}}: CS1 maint: unflagged free DOI (link)
  31. ^ a b Council, National Research (1969-12-31). In Situ Bioremediation: When Does it Work?. doi:10.17226/2131. ISBN 9780309048965.
  32. ^ a b Gkorezis, Panagiotis; Daghio, Matteo; Franzetti, Andrea; Hamme, Van; D, Jonathan; Sillen, Wouter; Vangronsveld, Jaco (2016-01-01). "The Interaction between Plants and Bacteria in the Remediation of Petroleum Hydrocarbons: An Environmental Perspective". Frontiers in Microbiology. 7: 1836. doi:10.3389/fmicb.2016.01836. ISSN 1664-302X. PMC 5116465. PMID 27917161.{{cite journal}}: CS1 maint: unflagged free DOI (link)
  33. ^ Azubuike, Christopher Chibueze; Chikere, Chioma Blaise; Okpokwasili, Gideon Chijioke (2016-11-01). "Bioremediation techniques–classification based on site of application: principles, advantages, limitations and prospects". World Journal of Microbiology and Biotechnology. 32 (11): 180. doi:10.1007/s11274-016-2137-x. ISSN 0959-3993. PMC 5026719. PMID 27638318.
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