Crude oil pollution is one of the most serious environmental issues today, and the clean-up procedure is perhaps the most difficult. Within one to three weeks, the vast majority of oil bacteria may degrade approximately 60% of the crude oil, leaving approximately 40% intact. The by-product metabolites produced during the breakdown of oil are essentially organic molecules in nature. These metabolites inhibit its enzymes, preventing the oil bacteria from further degrading the oil. By combining a variety of different oils with heterotrophic bacteria in a bioreactor, the rate of crude oil biodegradation was accelerated. In this study, two strains of oil-resistant, heterotrophic bacteria (OG1 and OG2-Erythrobacter citreus) and a bacterium that uses hydrocarbons (AR3-Pseudomonas pseudoalcaligenes) were used. Gas chromatography-mass spectroscopy was used to investigate the effectiveness of this consortium of symbiotic bacteria in the biodegradation of crude oil. According to gravimetric and gas chromatography analyses, the consortium bacteria digested 69.6% of the crude oil in the bioreactor, while the AR3 single strain was only able to destroy 61.9% of it. Under the same experimental conditions, consortium bacteria degraded approximately 84550.851 ppb (96.3%) of 16 aliphatic hydrocarbons and 9333.178 ppb (70.5%) of 16 aromatic hydrocarbons in the bioreactor. It may be inferred that the novel consortium of symbiotic bacteria accelerated the biodegradation process and had great potential for use in increasing the bioremediation of hydrocarbon-contaminated locations.
This study was undertaken to analyze the efficiency of Botryococcus sp. in the phycoremediation of domestic wastewater and to determine the variety of hydrocarbons derived from microalgal oil after phycoremediation. The study showed a significant (p < 0.05) reduction of pollutant loads of up to 93.9% chemical oxygen demand, 69.1% biochemical oxygen demand, 59.9% total nitrogen, 54.5% total organic carbon, and 36.8% phosphate. The average dry weight biomass produce was 0.1 g/L of wastewater. In addition, the dry weight biomass of Botryococcus sp. was found to contain 72.5% of crude oil. The composition analysis using Gas Chromatogram - Mass Spectrometry (GC-MS) found that phthalic acid, 2-ethylhexyltridecyl ester (C29H48O4), contributed the highest percentage (71.6%) of the total hydrocarbon compounds to the extracted algae oil. The result of the study suggests that Botryococcus sp. can be used for effective phycoremediation, as well as to provide a sustainable hydrocarbon source as a value-added chemical for the bio-based plastic industry.
Waxy crude oil is a problem to the oil and gas industry because wax deposition in pipelines reduces the quality of the crude oil. Currently, the industry uses chemicals to solve the problem but it is not environmentally friendly. As an alternative, the biodegradation approach is one of the options. Previously eleven thermophilic bacteria were isolated and exhibited high ability to degrade hydrocarbon up to 70% of waxy crude oil. However, despite the successful study on these single bacteria strains, it is believed that biodegradation of paraffin wax requires more than a single species. Five consortia were developed based on the biodegradation efficiency of 11 bacterial strains. Consortium 3 showed the highest biodegradation (77.77%) with more long-chain alkane degraded throughout the incubation compared to other consortia. Enhancement of hydrocarbon degradation was observed for all consortia especially in long chain alkane (C18-C40). Consortium 3 exhibited higher alkane monooxygenase, alcohol dehydrogenase, lipase, and esterase activities. Moreover, the dominant bacteria in the consortia were determined by denaturing gradient gel electrophoresis (DGGE), which showed the domination of genera Geobacillus, Parageobacillus, and Anoxybacillus. It can be concluded that the bacterial consortia showed higher biodegradation and improved degrading more long-chain hydrocarbon compared to a single isolate.
Treatment of petroleum-contaminated soil to a less toxic medium via physical and chemical treatment is too costly and requires posttreatment. This review focuses on the employment of phytoremediation and mycoremediation technologies in cleaning hydrocarbon-contaminated soil which is currently rare. It is considered environmentally beneficial and possibly cost-effective as it implements the synergistic interaction between plants and biosurfactant producing mycorrhiza to degrade hydrocarbon contaminants. This review also covers possible sources of hydrocarbon pollution in water and soil, toxicity effects, and current technologies for hydrocarbon removal and degradation. In addition to these problems, this review also discusses the challenges and opportunities of transforming the resultant treated sludge and treating plants into potential by-products for a higher quality of life for future generations.
This study investigated the optimum conditions for total petroleum hydrocarbon (TPH) removal from diesel-contaminated water using phytoremediation treatment with Scirpus grossus. In addition, TPH removal from sand was adopted as a second response. The optimum conditions for maximum TPH removal were determined through a Box-Behnken Design. Three operational variables, i.e. diesel concentration (0.1, 0.175, 0.25% Vdiesel/Vwater), aeration rate (0, 1 and 2 L/min) and retention time (14, 43 and 72 days), were investigated by setting TPH removal and diesel concentration as the maximum, retention time within the given range, and aeration rate as the minimum. The optimum conditions were found to be a diesel concentration of 0.25% (Vdiesel/Vwater), a retention time of 63 days and no aeration with an estimated maximum TPH removal from water and sand of 76.3 and 56.5%, respectively. From a validation test of the optimum conditions, it was found that the maximum TPH removal from contaminated water and sand was 72.5 and 59%, respectively, which was a 5 and 4.4% deviation from the values given by the Box-Behnken Design, providing evidence that S. grossus is a Malaysian native plant that can be used to remediate wastewater containing hydrocarbons.
Two types of flow system, free surface flow (FSF) and sub-surface flow (SSF), were examined to select a better way to remove total petroleum hydrocarbons (TPH) using diesel as a hydrocarbon model in a phytotoxicity test to Scirpus grossus. The removal efficiencies of TPH for the two flow systems were compared. Several wastewater parameters, including temperature (T, °C), dissolved oxygen (DO, mgL(-1)), oxidation-reduction potential (ORP, mV), and pH were recorded during the experimental runs. In addition, overall plant lengths, wet weights, and dry weights were also monitored. The phytotoxicity test using the bulrush plant S. grossus was run for 72 days with different diesel concentrations (1%, 2%, and 3%) (Vdiesel/Vwater). A comparison between the two flow systems showed that the SSF system was more efficient than the FSF system in removing TPH from the synthetic wastewater, with average removal efficiencies of 91.5% and 80.2%, respectively. The SSF system was able to tolerate higher diesel concentrations than was the FSF system.
In this study, we isolated two indigenous hydrocarbon-degrading bacteria from tarball found in Rhu Sepuluh beach, Terengganu, Malaysia. These bacteria were identified based on their physiological characteristic and 16S rRNA gene sequence analysis, and they showed 99% similarity with Cellulosimicrobium cellulans DSM 43879 and Acinetobacter baumannii ATCC 19606 respectively. Their hydrocarbon-degrading capabilities were tested using diesel-oil as sole carbon source. Results analysed using GC-MS, showed diesel-oil alkanes were degraded an average 64.4% by C. cellulans and 58.1% by A. baumannii with medium optical density reaching 0.967 (C. cellulans) and 1.515 (A. baumannii) in minimal salt media at 32°C for 10days. Individual diesel-oil alkanes were degraded between 10%-95.4% by C. cellulans and 0.2%-95.9% by A. baumannii. Both strains utilized diesel-oil for growth. The study suggests both strains are part of indigenous hydrocarbon-degrading bacteria in tarball with potential for bioremediation of oil-polluted marine environment.
The unique cellular enzymatic machinery of halophilic microbes allows them to thrive in extreme saline environments. That these microorganisms can prosper in hypersaline environments has been correlated with the elevated acidic amino acid content in their proteins, which increase the negative protein surface potential. Because these microorganisms effectively use hydrocarbons as their sole carbon and energy sources, they may prove to be valuable bioremediation agents for the treatment of saline effluents and hypersaline waters contaminated with toxic compounds that are resistant to degradation. This review highlights the various strategies adopted by halophiles to compensate for their saline surroundings and includes descriptions of recent studies that have used these microorganisms for bioremediation of environments contaminated by petroleum hydrocarbons. The known halotolerant dehalogenase-producing microbes, their dehalogenation mechanisms, and how their proteins are stabilized is also reviewed. In view of their robustness in saline environments, efforts to document their full potential regarding remediation of contaminated hypersaline ecosystems merits further exploration.
Greenhouse experiments were carried out to determine the phytotoxic effects on the plant Ludwigia octovalvis in order to assess its applicability for phytoremediation gasoline-contaminated soils. Using plants to degrade hydrocarbons is a challenging task. In this study, different spiked concentrations of hydrocarbons in soil (1, 2, and 3 g/kg) were tested. The results showed that the mean efficiency of total petroleum hydrocarbon (TPH) removal over a 72-day culture period was rather high. The maximum removal of 79.8 % occurred for the 2 g/kg concentration, while the removal rate by the corresponding unplanted controls was only (48.6 %). The impact of gasoline on plants included visual symptoms of stress, yellowing, growth reduction, and perturbations in the developmental parameters. The dry weight and wet weight of the plant slightly increased upon exposure to gasoline until day 42. Scanning electron microscopy (SEM) indicated change to the root and stem structure in plant tissue due to the direct attachment with gasoline contaminated compared to the control sample. The population of living microorganisms in the contaminated soil was found to be able to adapt to different gasoline concentrations. The results showed that L. octovalvis and rhizobacteria in gasoline-contaminated soil have the potential to degrade organic pollutants.
A novel kinetic model has been developed to explain the degradation of total petroleum hydrocarbons. Microbiome engineered biochar amendment may result in a synergistic impact on degradation of total petroleum hydrocarbons (TPHs). Therefore, the present study analyzed the potential of hydrocarbon-degrading bacteria A designated as Aeromonas hydrophila YL17 and B as Shewanella putrefaciens Pdp11 morphological characterized as rod shaped, anaerobic and gram-negative immobilized on biochar, and the degradation efficiency was measured by gravimetric analysis and gas chromatography-mass spectrometry (GC-MS). Whole genome sequencing of both strains revealed the existence of genes responsible for hydrocarbon degradation. In 60 days remediation setup, the treatment consisting of immobilization of both strains on biochar proved more efficient with less half-life and better biodegradation potentials compared to biochar without strains for decreasing the content of TPHs and n-alkanes (C12-C18). Enzymatic content and microbiological respiration showed that biochar acted as a soil fertilizer and carbon reservoir and enhanced microbial activities. The removal efficiency of hydrocarbons was found to be a maximum of 67% in soil samples treated with biochar immobilized with both strains (A + B), followed by biochar immobilized with strain B 34%, biochar immobilized with strain A 29% and with biochar 24%, respectively. A 39%, 36%, and 41% increase was observed in fluorescein diacetate (FDA) hydrolysis, polyphenol oxidase and dehydrogenase activities in immobilized biochar with both strains as compared to control and individual treatment of biochar and strains. An increase of 35% was observed in the respiration rate with the immobilization of both strains on biochar. While a maximum colony forming unit (CFU/g) was found 9.25 with immobilization of both strains on biochar at 40 days of remediation. The degradation efficiency was due to synergistic effect of both biochar and bacteria based amendment on the soil enzymatic activity and microbial respiration.
Hydrocarbon-degrading bacteria, which can be found living with eukaryotic phytoplankton, play a pivotal role in the fate of oil spillage to the marine environment. Considering the susceptibility of calcium carbonate-bearing phytoplankton under future ocean acidification conditions and their oil-degrading communities to oil exposure under such conditions, we investigated the response of non-axenic E. huxleyi to crude oil under ambient versus elevated CO2 concentrations. Under elevated CO2 conditions, exposure to crude oil resulted in the immediate decline of E. huxleyi, with concomitant shifts in the relative abundance of Alphaproteobacteria and Gammaproteobacteria. Survival of E. huxleyi under ambient conditions following oil enrichment was likely facilitated by enrichment of oil-degraders Methylobacterium and Sphingomonas, while the increase in relative abundance of Marinobacter and unclassified Gammaproteobacteria may have increased competitive pressure with E. huxleyi for micronutrient acquisition. Biodegradation of the oil was not affected by elevated CO2 despite a shift in relative abundance of known and putative hydrocarbon degraders. While ocean acidification does not appear to affect microbial degradation of crude oil, elevated mortality responses of E. huxleyi and shifts in the bacterial community illustrates the complexity of microalgal-bacterial interactions and highlights the need to factor these into future ecosystem recovery projections.
This study aimed to isolate biosurfactant-producing and hydrocarbon-degrading actinomycetes from different soils using glycerol-asparagine and starch-casein media with an antifungal agent. The glycerol-asparagine agar exhibited the highest number of actinomycetes, with a white, low-opacity medium supporting pigment production and high growth. Biosurfactant analyses, such as drop collapse, oil displacement, emulsification, tributyrin agar test, and surface tension measurement, were conducted. Out of 25 positive isolates, seven could utilize both olive oil and black oil for biosurfactant production, and only isolate RP1 could produce biosurfactant when grown in constrained conditions with black oil as the sole carbon source and inducer, demonstrating in situ bioremediation potential. Isolate RP1 from oil-spilled garden soil is Gram-staining-positive with a distinct earthy odor, melanin formation, and white filamentous colonies. It has a molecular size of ~621 bp and 100% sequence similarity to many Streptomyces spp. Morphological, biochemical, and 16 S rRNA analysis confirmed it as Streptomyces sp. RP1, showing positive results in all screenings, including high emulsification activity against kerosene (27.2%) and engine oil (95.8%), oil displacement efficiency against crude oil (7.45 cm), and a significant reduction in surface tension (56.7 dynes/cm). Streptomyces sp. RP1 can utilize citrate as a carbon source, tolerate sodium chloride, resist lysozyme, degrade petroleum hydrocarbons, and produce biosurfactant at 37°C in a 15 mL medium culture, indicating great potential for bioremediation and various downstream industrial applications with optimization.
Toxic inorganic and organic chemicals are major contributors to environmental contamination and pose major health risks to human population. In this work, Dracaena reflexa and Podocarpus polystachyus were investigated for their potential to remove hydrocarbons from 2.5% and 1% diesel fuel-contaminated soil amended individually with 5% organic wastes (tea leaf, soy cake and potato skin) for a period of 270 days. Loss of 90% and 99% oil was recorded in soil contaminated with 2.5% and 1% oil with soy cake amendment, respectively, compared with 52% and 62% in unamended soil with D. reflexa at the end of 270 days. Similarly, 84% and 91% oil loss was recorded for P. polystachyus amended with organic wastes in 2.5% and 1% oil, respectively. Diesel fuel disappeared more rapidly in the soil amendment with SC than in other organic waste supplementation. It was evident that plants did not accumulate hydrocarbon from the soil, while the number of hydrocarbon-utilizing bacteria was high in the rhizosphere, thus suggesting that the mechanism of the oil degradation was rhizodegradation. The kinetic model result indicated a high rate of degradation in soil amendment with SC at 1% with D. reflexa compared with other treatments. Thus, a positive relationship was observed between diesel hydrocarbon degradation with plant biomass production. Dracaena reflexa with organic wastes amendment has a greater potential of restoring hydrocarbon-contaminated soil compared to P. polystachyus plant.
Major degraders of petroleum hydrocarbons in tropical seas have been indicated only by laboratory culturing and never through observing the bacterial community structure in actual environments. To demonstrate the major degraders of petroleum hydrocarbons spilt in actual tropical seas, indigenous bacterial community in seawater at Sentosa (close to a port) and East Coast Park (far from a port) in Singapore was analyzed. Bacterial species was more diverse at Sentosa than at the Park, and the composition was different: γ-Proteobacteria (57.3%) dominated at Sentosa, while they did not at the Park. Specialized hydrocarbonoclastic bacteria (SHCB), which use limited carbon sources with a preference for petroleum hydrocarbons, were found as abundant species at Sentosa, indicating petroleum contamination. On the other hand, SHCB were not the abundant species at the Park. The abundant species of SHCB at Sentosa were Oleibacter marinus and Alcanivorax species (strain 2A75 type), which have previously been indicated by laboratory culturing as important petroleum-aliphatic-hydrocarbon degraders in tropical seas. Together with the fact that SHCB have been identified as major degraders of petroleum hydrocarbons in marine environments, these results demonstrate that the O. marinus and Alcanivorax species (strain 2A75 type) would be major degraders of petroleum aliphatic hydrocarbons spilt in actual tropical seas.
In this study, bulrush (Scirpus grossus) was subjected to a 72 day phytotoxicity test to assess its ability to phytoremediate diesel contamination in simulated wastewater at different concentrations (0, 8700, 17,400 and 26,100mg/L). Diesel degradation by S. grossus was measured in terms of total petroleum hydrocarbon (TPH-D). The TPH-D concentration in the synthetic wastewater was determined with the liquid-liquid extraction method and gas chromatography. S. grossus was found to reduce TPH-D by 70.0 and 80.2% for concentrations of 8700 mg/L and 17,400mg/L, respectively. At a diesel concentration of 26,100mg/L, S. grossus died after 14 days. Additionally, the biomass of S. grossus plants was found to increase throughout the phytotoxicity test, confirming the ability of the plant to survive in water contaminated with diesel at rates of less than 17,400mg/L.
Weathered crude oil (WCO) removals in shoreline sediment samples were monitored for 60 days in bioremediation experimentation. Experimental modeling was carried out using statistical design of experiments. At optimum conditions maximum of 83.13, 78.06 and 69.92% WCO removals were observed for 2, 16 and 30 g/kg initial oil concentrations, respectively. Significant variations in the crude oil degradation pattern were observed with respect to oil, nutrient and microorganism contents. Crude oil bioremediation were successfully described by a first-order kinetic model. The study indicated that the rate of hydrocarbon biodegradation increased with decrease of crude oil concentrations.
This study examined the capacity of immobilized bacteria to degrade petroleum hydrocarbons. A mixture of hydrocarbon-degrading bacterial strains was immobilized in alginate and incubated in crude oil-contaminated artificial seawater (ASW). Analysis of hydrocarbon residues following a 30-day incubation period demonstrated that the biodegradation capacity of the microorganisms was not compromised by the immobilization. Removal of n-alkanes was similar in immobilized cells and control cells. To test reusability, the immobilized bacteria were incubated for sequential increments of 30 days. No decline in biodegradation capacity of the immobilized consortium of bacterial cells was noted over its repeated use. We conclude that immobilized hydrocarbon-degrading bacteria represent a promising application in the bioremediation of hydrocarbon-contaminated areas.
Due to the increasing importance of diesel and petroleum for industrial development during the last century, petrochemical effluents have significantly contributed to the pollution of aquatic and soil environments. The contamination generated by petroleum hydrocarbons can endanger not only humans but also the environment. Phytoremediation or plant-assisted remediation can be considered one of the best technologies to manage petroleum product-contaminated water and soil. The main advantages of this method are that it is environmentally-friendly, potentially cost-effective and does not require specialised equipment. The scope of this review includes a description of hydrocarbon pollutants from petrochemical industries, their toxicity impacts and methods of treatment and degradation. The major emphasis is on phytodegradation (phytotransformation) and rhizodegradation since these mechanisms are the most favourable alternatives for soil and water reclamation of hydrocarbons using tropical plants. In addressing these issues, this review also covers challenges to retrieve the environment (soil and water) from petroleum contaminations through phytoremediation, and its opportunities to remove or reduce the negative environmental impacts of petroleum contaminations and restore damaged ecosystems with sustainable ways to keep healthy life for the future.
The coral reefs of the Persian Gulf are the most diverse systems of life in the marine environment of the Middle East. Unfortunately, they are highly threatened by local and global stressors, particularly oil pollutants. This is the first quantitative and qualitative study aimed at assessing the concentration and sources of n-alkanes and POPs (PAHs, PCBs and PCNs) in coral tissues, symbiotic algae (zooxanthellae), reef sediments and seawaters in coral reefs of Lark and Kharg in the Persian Gulf, Iran. This work was conducted on eight species of six genera and three families of hard corals and one family of soft coral. A significant variation in the concentration of ∑30n-alkanes and POPs (∑40PAHs, ∑22PCBs and 20PCNs) was found in the decreasing order: zooxanthellae > coral tissue > skeleton > reef sediment > seawater. The bioaccumulation of these compounds was 2-times higher in ahermatypic than in hermatypic corals, among which significant variations were observed in both sites. In Kharg, Porites lutea had the highest mean concentration of ∑30n-alkanes and ∑40PAHs in soft tissue, whereas the lowest values were in Platygyra daedalea. A contrasting trend was documented for ∑22PCBs and 20PCNs, with the highest level reported in soft tissue of P. daedalea and the lowest in P. lutea at Kharg. Compositional pattern of AHs and PAHs demonstrated the predominance of LMW-PAHs and n-alkanes. In skeleton and reef sediments, tetra, penta and tri-CBs were the most abundant PCBs congeners followed by di-CB > hexa-CB > hepta-CB > octa-CB,whiletri-CB > di-CB > tetra-CB > penta-CB > hexa-CB > hepta-CB > octa-CB was observed for soft tissue, zooxanthellae and seawater. The results of RAD test indicated significantly negative correlation between total concentration of these compounds with zooxanthellae density, the chlorophyll-a and C2 in corals at both reefs. This is the first report on levels, health assessment and source apportionments of POPs in zooxanthellae and a first step in the implementation of specific coral reef management measures.
Soil contamination by hydrocarbons, especially by used lubricating oil, is a growing problem in developing countries, which poses a serious threat to the environment. Phytoremediation of these contaminated soils offers environmental friendly and a cost effective method for their remediation. Hibiscus cannabinus was studied for the remediation of soil contaminated with 2.5 and 1% used lubricating oil and treated with organic wastes [banana skin (BS), brewery spent grain (BSG) and spent mushroom compost (SMC)] for a period of 90 days under natural conditions. Loss of 86.4 and 91.8% used lubricating oil was recorded in soil contaminated with 2.5 and 1% oil and treated with organic wastes respectively at the end of 90 days. However, 52.5 and 58.9% oil loss was recorded in unamended soil contaminated with 2.5 and 1% oil, respectively. The plant did not accumulate hydrocarbon from the soil but shows appreciable accumulation of Fe and Zn in the root and stem of H. cannabinus at the end of the experiment. The first order kinetic rate of uptake of Fe and Zn in H. cannabinus was higher in organic wastes amendment treatments compared to the unamended treatments, which are extremely low. The results of this study suggest that H. cannabinus has a high potential for remediation of hydrocarbon and heavy metal contaminated soil.