Oil Bioremediation Research Articles

Bioremediation of hydrocarbon pollutants by Pseudomonas putida under optimal conditions

Background: The extreme toxicity of petroleum products to human and environmental health, particularly when linked to large-scale unintentional spills, has made them one of the most serious and current environmental pollution issue. Petroleum products are a complex mixture of hydrocarbons.Methods: Oil-contaminated soil samples were gathered and utilized to isolate hydrocarbon-degrading bacterial isolates. The bacterial isolates possessing bioremediation capabilities were identified using phenotypic determination and biochemical properties. The Optimal conditions including incubation period, temperature, different pH values and different carbon sources were studied for bioremediation of crude oil by bacterial isolates.Result: In Wasit province of Iraq, Pseudomonas putida, P. fluorescens, and P. aeruginosa isolated from soil samples tainted with petroleum hydrocarbons. Pseudomonas putida have more ability for bioremediation of crude oil (68%) compared to P. aeruginosa and P. fluorescens (47%, 58%) respectively. The results of optical density (600nm), biomass and (E24%) index for Pseudomonas putida were (1.080, 1.98 and 60%) respectively. The Optimal conditions (incubation period, temperature, different pH values and different carbon sources) were studied for bioremediation of crude oil by Pseudomonas putida by using liquid BHM supplemented with 1% crude oil. The result of this study showed that the incubation period of 9 days was the optimum for bioremediation of hydrocarbons which was 88.33%. The optimum temperature and pH were 35 °C and 7 respectively. The carbon source (glucose and fructose) was optimal for hydrocarbon bioremediation.Conclusion: This bacterial isolate Pseudomonas putida can be use in petroleum hydrocarbon bioremediation process.Keywords: Pseudomonas putida; Bioremediation; Optical density

Optimization of Bioremediation of Crude Oil Polluted Soil through Variation in Moisture Content and Proportion of Augmenting Bacterium and Fungus

Aim: The aim of this study was to enhance bioremediation of crude-oil polluted soil through optimization procedures. Study Design: The Box-Benkhen coded values format and experimental design were used in the design of the study. Place and Duration of Study: The study was carried out in the Microbiology laboratory of the Department of Microbiology, Rivers State University, Nigeria, between February 2024 and July 2024. Methodology: Hydrocarbon utilizing bacteria (HUB) & fungi (HUF) were isolated from crude-oil polluted soil, and screened for hydrocarbon degrading ability (HCDA). HUB and HUF with the highest HCDA were identified and used for bioremediation optimization experiment. In the experiment, different combination of moisture content (MC), and proportion of HUB (pHUB) & HUF (pHUF) were investigated for their effect on extent of hydrocarbon reduction (EoHR). EoHR obtained from the different combinations were fitted using a generalized polynomial model so as to obtain a polynomial equation for predicting EoHR. The equation was used in generating prediction profiles from which the combined values of the 3 parameters that will lead to the highest EoHR was determined. The predicted combined values were implemented in a new setup. A control and a setup enhanced with fertilizer were also prepared. The setups were maintained for 3 weeks. On day 0, 14, and 21 samples were collected and analyzed for total hydrocarbon concentration (THC). Results: The results obtained showed that among the coded isolated HUB and HUF, HB5 and HF1 had the highest HCDA; 36.4% and 4.1% respectively, and were identified phenotypically as Klebsiella ornithinolytica and Aspergillus flavus respectively. The results of the optimization experiment and prediction profiles showed that the highest (68.6%) EoHR was achievable at MC = 20%, pHUB = 10%, and pHUF = 1%. The actual EoHR on day 21 in the optimized, enhanced optimized, and control setups were 60.3, 58.9, and 39.9% respectively. Conclusion: Bioremediation optimization studies is advantageous and should be carried out on crude-oil polluted sites before carrying out bioremediation.

Crude oil degrading efficiency of formulated consortium of bacterial strains isolated from petroleum-contaminated sludge.

Crude oil contamination has been widely recognized as a major environmental issue due to its various adverse effects. The use of inhabitant microorganisms (native to the contaminated sites) to detoxify/remove pollutants owing to their diverse metabolic capabilities is an evolving method for the removal/degradation of petroleum industry contaminants. The present study deals with the exploitation of native resident bacteria from crude oil contaminated site (oil exploration field) for bioremediation procedures. Fifteen (out of forty-four) bioremediation-relevant aerobic bacterial strains, belonging to the genera of Bacillus, Stenotrophomonas, Pseudomonas, Paenibacillus, Rhizobium, Burkholderia, and Franconibacter, isolated from crude oil containing sludge, have been selected for the present bioremediation study. Crude oil bioremediation performance of the selected bacterial consortium was assessed using microcosm-based studies. Stimulation of the microbial consortium with nitrogen or phosphorous led to the degradation of 60-70% of total petroleum hydrocarbon (TPH) in 0.25% and 0.5% crude oil experimental sets. CO2 evolution, indicative of crude oil mineralization, was evident with the highest evolution being 28.6mgmL-1. Ecotoxicity of treated crude oil-containing media was assessed using plant seed germination assay, in which most of the 0.25% and 0.5% treated crude oil sets gave positive results thereby suggesting a reduction in crude oil toxicity.

Bioremediation of Oil and Natural Gas Industry Waste Using Methanogens: Current Status and Future Perspective to Biohythane Production

Bioremediation of Oil and Natural Gas Industry Waste Using Methanogens: Current Status and Future Perspective to Biohythane Production

Differential biodegradation of alkanes in crude oil by three oleophilic strains

Ex situ bioremediation of crude oil in bioreactors by oleophilic bacteria permits to regulate temperature, nutrients and oxygen and the results in petroleum removal are much more effective than in situ bioremediation. This article quantify the biodegradation of alkanes which conform crude oil in bioreactors by three oleophilic strains. Samples were collected from the bioreactors and extracted in dichloromethane (10:1 aqueous/organic v/v) for amplification in GC-FID analysis of individual alkanes. 94% of crude oil (initial concentration 8.6 g l−1) is removed in 24 days by Bacillus licheniformis and in 38 days by Pseudomonas putida and Paenibacillus glucanolyticus. Higher biodegradation rates (9.0–26.5 mg l−1d−1) are reached for the lightest fraction of crude oil by Bacillus licheniformis (C10–C16), and much lower for C22–C25 alkanes (4.4–8.9 mg l−1d−1). Pseudomonas putida and Paenibacillus glucanolyticus degrade a wide range of alkanes at 1.6–9.6 mg l−1d−1, with preference over C8–C16 hydrocarbons. Heptane and octane (alkanes cited to be toxic) are difficult for biodegradation in two strains (B. licheniformis and P. putida) and a recalcitrant “window” C18–C21 is observed for the three oleophilic strains in the alkane range studied.

Bioremediation of crude oil polluted surface water using specialised alginate-based nanocomposite beads loaded with hydrocarbon-degrading bacteria and inorganic nutrients

This study explored the elimination of petroleum from river water using matrix-bound composites of iron oxide nanoparticles (IONPs) decorated on biochar with immobilized hydrocarbon-degrading bacteria (HCB) and monoammonium phosphate (MAP) at 10%v/v pollution levels. The beads containing biochar and IONPs (BCNP) showed better pollutant removal than those with biochar alone (BC). The treatment with beads containing biochar, IONPs, HCB and MAP had the highest removal efficiency for TPH (98.5%) and PAH (93.7%) and the lowest biodegradation half-lives. The removal efficiency for TPH and PAH was greater in the treatment microcosms than in the controls with significant differences (p ≤ 0.05) between the two groups. TPH and PAH elimination levels differed significantly (p ≤ 0.05) between systems with the BC bead and those with the BCNP beads. The study revealed a steady increase in the abundance of the total cultivable heterotrophic bacteria and cultivable hydrocarbon utilizing bacteria in the treatment microcosms after an initial decline, with strong negative correlation between bacterial abundance and TPH and PAH concentrations based on r values (0.5 − 1.0) and shared variances obtained. Bacillus, Pseudomonas and Klebsiella pneumoniae dominated as the drivers of the bioremediation process while organic acids and esters (25.0%) and ketones (14.3%) were the main biodegradation metabolite groups.

Using PyCaret to model Chlorella vulgaris's growth response to salinity and oil contamination for crude oil bioremediation

ABSTRACT Crude oil spills significantly impact aquatic ecosystems, necessitating innovative remediation strategies. Microalgae-based bioremediation, particularly with Chlorella vulgaris, offers a promising solution. This study introduces a novel framework that evaluates the combined effects of selected environmental stressors on microalgal adaptability, advancing beyond traditional isolated factor analyses. By integrating a factorial experimental design with a machine learning approach using PyCaret AutoML and SHAP values, we provide a detailed examination of how crude oil concentration, salinity, and exposure duration affect C. vulgaris growth. The Extra Trees Regressor model emerged as highly accurate in predicting biomass concentration, a crucial adaptability indicator, achieving an MAE of 0.0202, RMSE of 0.029, and an R² of 0.8875. SHAP analysis highlighted salinity and crude oil as significant growth influencers, with exposure duration playing a minor role. Notably, C. vulgaris exhibited more sensitivity to salinity than to crude oil, indicating potential high-salinity challenges but also a strong tolerance to oil pollutants. These findings enhance our understanding of microalgal responses in polluted environments and suggest improved bioremediation approaches for saline waters affected by oil spills, leveraging the synergy of environmental factors and machine learning insights.

The use of pigeon droppings in bioremediation of crude oil contaminated open farm

The use of pigeon droppings in bioremediation of crude oil contaminated open farm

Microbial remediation of petroleum-contaminated soil focused on the mechanism and microbial response: a review.

The environmental pollution caused by petroleum hydrocarbons has received considerable attention in recent years. Microbial remediation has emerged as the preferred method for the degradation of petroleum hydrocarbons, which is experiencing rapid development driven by advancements in molecular biology. Herein, the capacity of different microorganisms used for crude oil bioremediation was reviewed. Moreover, factors influencing the effectiveness of microbial remediation were discussed. Microbial remediation methods, such as bioaugmentation, biostimulation, and bioventilation, are summarized in this review. Aerobic and anaerobic degradation mechanisms were reviewed to elucidate the metabolic pathways involved. The impacts of petroleum hydrocarbons on microorganisms and the environment were also revealed. A brief overview of synthetic biology and a unique perspective of technique combinations were presented to provide insight into research trends. The challenges and future outlook were also presented to stimulate contemplation of the mechanisms involved and the development of innovative techniques.

Bioremediation of Crude Oil Contaminated Saline Soil Using a Bacterial Consortium and Different Carriers

AbstractBioremediation of crude‐oil‐contaminated soils is critical to supporting soil ecosystem health and functioning. Here, to explore the potential for different remediation strategies to restore soil microbial functioning and enhance remediation, three strains of bacteria, Bacillus toyonensis, Bacillus sp. and Bacillus cereus, were inoculated to saline, crude oil‐contaminated soil collected from Azadegan Oil Field, Iran. Bacteria were added using different carriers, including corn biochar, humic acid, chitosan and kaolin, and free bacteria as a control. Changes in soil metal concentrations and residual crude oil were determined after 60 days of incubation. Soil respiration, microbial biomass, and enzyme (dehydrogenase and catalase) activities were measured. The carriers accompanying a bacterial consortium were more efficient than free bacteria in removing crude oil and heavy metals from soil, and they performed better in removing n‐alkanes with carbon chains of C12–24. The oil removal rate and metal immobilization efficiencies were greatest in soils with chitosan‐bacteria treatment, with removal of crude oil, Al, Pb, Sr, and Cr (93%, 63%, 37%, 54%, and 15%, respectively) greater than for soils with free cells without treatment (61%, 54%, 9%, 50%, and 3%, respectively). Soil electrical conductivity decreased after application of bacterial mixtures with and without carriers. The inoculation of bacteria and application of soil amendments increased soil microbial biomass carbon and the activity of catalase and dehydrogenase. Overall, our results indicated the greatest potential remediation was achieved by applying immobilized microorganisms in chitosan to accelerate the biodegradation of crude oil and the removal of heavy metals.

Exploring Hydrocarbon Degradation Capacity and Mathematical Modeling of Mixotrophic Growth Kinetics in Tetradesmus obliquus and Desmodesmus armatus for Crude Oil Bioremediation

Exploring Hydrocarbon Degradation Capacity and Mathematical Modeling of Mixotrophic Growth Kinetics in Tetradesmus obliquus and Desmodesmus armatus for Crude Oil Bioremediation

POTENTIAL OF MYCORRHIZAL INOCULATION AND CATTLE RUMEN DIGESTA IN THE BIOREMEDIATION OF SPENT ENGINE OIL CONTAMINATED SOIL

Soil pollution by crude oil contamination has become a major constraint on agricultural productivity. Physicochemical techniques are often expensive. However, bioremediation of petroleum hydrocarbon polluted soil is cost-effective. Therefore, the study was carried out to analyze the influence of mycorrhiza and cattle rumen digesta on bioremediation of Spent Engine Oil (SEO) contaminated soil in Dutse, Jigawa state. Soil samples were randomly collected from the University Research and Teaching Farm. About 2.5 kg of sterilized topsoil (0–15 cm) was filled into pots and arranged in a 2×2×3 factorial experiment in completely randomized design with three replications. Mycorrhiza and cattle rumen digesta were at two levels, while SEO was at three levels. Data were collected on the total petroleum hydrocarbon (TPH) content, bacterial and fungal colony count. Data were analyzed using ANOVA at α 0.05. Results obtained from the study show that mycorrhiza and cattle rumen digesta increased the colonies of fungi and bacteria resulting in significantly enhanced TPH degradation in the contaminated soil. However, cattle rumen digesta significantly (p<0.05) enhanced TPH degradation, bacterial and fungal population the most compared to mycorrhiza alone. Combined cattle rumen digesta and mycorrhiza application resulted in significantly (p<0.05) lower residual TPH content in the contaminated soil compared to using cattle rumen digesta or mycorrhiza alone. Thus, cattle rumen digesta and mycorrhiza should be used in bioremediation of petroleum hydrocarbon impacted soils.

Bioremediation of diesel oil polluted seawater by a hydrocarbon-degrading bacterial consortium with oleophilic nutrients

The use of oleophilic nutrients that contained nitrogen and phosphorus is a valid strategy to enhance bioremediation rate in the open marine environments contaminated with hydrocarbons where the presence of nutrients is limited. The bioremediation effectiveness of the natural oleophilic nutrients (uric acid and soya bean lecithin) and an artificial oleophilic fertilizer prepared in this study were tested with an efficient bacterial consortium. The artificial oleophilic fertilizer was prepared using urea solution, soya bean lecithin, alcohol and oleic acid to form a water-in-oil type microemulsion system. The bioremediation potential of the oil-degrading bacterial consortium and these oleophilic nutrients were implemented by flask-shaking tests and laboratory mesocosm experiments. The diesel oil degradation ratios of the natural oleophilic nutrients and oleophilic fertilizer were up to 73% and 60%, respectively, at the optimal application rate during the shake flask trials. In the mesocosm experiments, the oil concentration of the oleophilic fertilizer + bacterial consortium group was decreased from about 8000 mg/L to 823 mg/L after six days of incubation, and the number of bacteria in the seawater increased from 3 × 104 to 1.8 × 1010 CFU/mL. The combination of these oleophilic nutrients with and the consortium was an effective strategy to enhance bioremediation rate. This method could be exploited further for the development of an effective bioremediation technology for the marine oil pollution.

Isolation and characterization of Candida tropicalis B: a promising yeast strain for biodegradation of petroleum oil in marine environments

The increasing interest in environmental protection laws has compelled companies to regulate the disposal of waste organic materials. Despite efforts to explore alternative energy sources, the world remains heavily dependent on crude petroleum oil and its derivatives. The expansion of the petroleum industry has significant implications for human and environmental well-being. Bioremediation, employing living microorganisms, presents a promising approach to mitigate the harmful effects of organic hydrocarbons derived from petroleum. This study aimed to isolate and purify local yeast strains from oil-contaminated marine water samples capable of aerobically degrading crude petroleum oils and utilizing them as sole carbon and energy sources. One yeast strain (isolate B) identified as Candida tropicalis demonstrated high potential for biodegrading petroleum oil in seawater. Physiological characterization revealed the strain’s ability to thrive across a wide pH range (4–11) with optimal growth at pH 4, as well as tolerate salt concentrations ranging from 1 to 12%. The presence of glucose and yeast extract in the growth medium significantly enhanced the strain's biomass formation and biodegradation capacity. Scanning electron microscopy indicated that the yeast cell diameter varied based on the medium composition, further emphasizing the importance of organic nitrogenous sources for initial growth. Furthermore, the yeast strain exhibited remarkable capabilities in degrading various aliphatic and aromatic hydrocarbons, with a notable preference for naphthalene and phenol at 500 and 1000 mg/l, naphthalene removal reached 97.4% and 98.6%, and phenol removal reached 79.48% and 52.79%, respectively. Optimization experiments using multi-factorial sequential designs highlighted the influential role of oil concentration on the bioremediation efficiency of Candida tropicalis strain B. Moreover, immobilized yeast cells on thin wood chips demonstrated enhanced crude oil degradation compared to thick wood chips, likely due to increased surface area for cell attachment. These findings contribute to our understanding of the potential of Candida tropicalis for petroleum oil bioremediation in marine environments, paving the way for sustainable approaches to address oil pollution.

Enhancing Bioremediation of Crude Oil Contaminated Soil from Kokori, Delta State, Nigeria Using Cow Dung as Biostimulant

This study was aimed at assessing the bioremediation of crude oil contaminated soil from kokori, Delta state, Nigeria by indigenous bacteria flora using cow dung as biostimulants. The activities of commercial agricultural investors, small-scale farmers, and the indigenous people of the Nigerian Niger Delta, whose livelihood has been heavily dependent on agriculture, have all been severely impeded and even destroyed by the crude oil spill. Therefore, a solution to the issue of oil spills in these places is required. Hydrocarbon contaminated soil was collected from a crude oil polluted sites and a non-contaminated soil was collected from Agricultural cultivable farmland in kokori, Delta state and its physicochemical properties were determined before the bioremediation period. The nutrients composition of cow dung used for the study was determined. After adding different amounts of cow dung to 200g of soil, the mixture was left to incubate at room temperature for six (6) weeks. Throughout the course of the experiment, measurements of the total petroleum hydrocarbon eliminated and the hydrocarbon-utilizing bacteria count (HUB) were made at one-week intervals. Increased in the HUB counts with corresponding decrease in the amount of hydrocarbons were observed. While highest HUB count was observed for the various amendment at week six, 5g/kg cow dung treatments had the highest count (3.35 x103cfu/g) with residual hydrocarbon (519.56mg/kg) while treatments 2.5g/kg cow dung had 2.91x103cfu/g with residual hydrocarbon of 541.06mg/kg. The result further revealed the degree of degradation likewise the bioremediation efficiency with C having the highest degree of degradation of (49.04%) at week six and bioremediation efficiency of (75.26%) at the first week of study. The introduction of cow dung into soil that has been contaminated with hydrocarbons promotes the growth of naturally occurring bacteria that break down hydrocarbons, leading to a greater amount of hydrocarbon degradation.

Crude oil biodegradation potential of lipase produced by Bacillus subtilis and Pseudomonas aeruginosa isolated from hydrocarbon contaminated soil

Microbial biodegradation of oil pollutants and their derivatives has become the most environmental-friendly method in the developing world. The aim of this study was to evaluate crude oil biodegradation potential of lipase produced by indigenous bacteria from oil contaminated soil. Indigenous bacteria isolates were identified as species of Bacillus subtilis and Pseudomonas aeruginosa, the isolates were able to produce lipase as revealed in their zone of clearance on tween 80 agar plates and the presence of lipase produced by the two bacteria were further confirmed using spectrophotometric analyses. Lipase produced by B. subtilis showed maximal lipase activity at pH 8 and 40°C while the enzyme produced by P. aeruginosa showed maximal lipase activity (U/mL) at pH 8 and 50°C when subjected to various pH and temperature respectively. Lipase produced by B. subtilis recorded 8.11 ± 0.70% of crude oil degradation in mineral salt medium within 28 days, while that of P. aeruginosa recorded 15.6 ± 0.03% of crude oil biodegradation. The GC–MS analysis of the crude oil treatment showed complete mineralization of several compounds, and also showed peak reduction which indicates lipase efficiency in the degradation of hydrocarbons. As revealed by GC–MS analysis, out of the 8 hydrocarbons identified in an undegraded oil, 5 were completely degraded by the enzyme activities while 2 (toluene and methyl, cyclopentane) were identified with hydrocarbons treated with lipase. The enzymes produced by B. subtilis and P. aeruginosa can serve as useful product for bioremediation of crude oil contaminated soil.

Tracking trash to treasure: in situ monitoring of single microbial cell oil biosynthesis from waste cooking oil using Raman spectroscopy and imaging.

Waste cooking oil is a major pollutant that contaminates terrestrial and aquatic bodies which is generated from household kitchens and eateries. The bioremediation of waste cooking oil (WCO) into microbial oil, also known as single microbial cell oil (SMCO), can be accomplished by oleaginous microbes. Conventional methods excel in SMCO analysis but lack efficacy for in situ or lysis-free monitoring of nascent SMCO synthesis and turnover. To bridge this knowledge gap, this study shows the applicability of Raman reverse stable isotope probing (RrSIP) in monitoring time-dependent nascent SMCO synthesis and assimilation in Yarrowia lipolytica, an oleaginous yeast grown in hydrophilic (glucose) as well as hydrophobic carbon sources (cooking oil and waste cooking oil). This study also combines the RrSIP approach with Raman imaging for temporal visualization of the distribution and turnover dynamics of the SMCO pool in a single cell. Our finding provides a unique perspective utilizing optical spectroscopy methods for lysis-free SMCO analysis and holds potential for prospective utility as an adjunct tool in bioprocess and biofuel industries.

Bioremediation of Crude Oil Polluted Soil by Cyanobacteria and Bacterial Consortium in Izombe Community of Imo State, Nigeria.

Bioremediation of Crude Oil Polluted Soil by Cyanobacteria and Bacterial Consortium in Izombe Community of Imo State, Nigeria.

Evaluation of Organic Treatments for Enhanced Bioremediation of Crude Oil Impacted Soil in Bayelsa State, Nigeria

The aim of this study was to evaluate organic treatments for enhanced bioremediation of crude oil impacted soil in Bayelsa State, Nigeria. Koroama community in Gbarain clan, Yenagoa local government area, Bayelsa State, Nigeria was randomly selected for the study. The experimental research design was adopted for this study, which was carried out for 28 days. The required soil samples were collected at a depth of 0 – 15cm from a 3 x 3m experimental plot developed in a farmland in Koroama community. Five sampled points, in the form of flat beds A, B, C, D and E respectively were randomly selected. Sampled beds A to D were simulated (contaminated) with 2.25kg of crude oil. The objective is to simulate conditions of a major crude oil spill. The crude oil contaminated soil samples were then allowed to condition for 6 days before treatment with 2kg organic treatments like goat manure (GM), poultry droppings (PD) and the combination of goat manure and poultry droppings (GM & PD). Bed E was unpolluted and untreated (control A) while Bed D was crude oil impacted and untreated soil (control B). The study showed that the impact of crude oil on the soil affects both the physical properties of the soil. The study also showed that the sampled organic treatments were all effective in restoring the physical properties of the crude oil impacted soil. At the end of the study, GM showed the highest TPH degradation rate (62.1%) followed by PD (57.1%) and the least, the combination of GM & PD (52.0%). Hence, the application of GM, PD and the combination of GM & PD are highly recommended for bioremediation of crude of impacted soil with special preference to GM.

Plant biomass extracted eco-friendly natural surfactant enhanced bio-electrokinetic remediation of crude oil contaminated soil

The current work investigates bioremediation (BIO) and electrokinetic (EK) remediation of crude oil hydrocarbons utilizing the biomass-electrokinetic (BIO-EK) approaches. The use of natural surfactants derived from plant biomass may improve remediation capacity by enhancing the solubility of organic pollutants. Sapindus mukorossi, a natural surfactant producer, was extracted from plant biomass in this study. The crude oil biodegradation efficiency was reported to be 98 %. In nature, FTIR confirms that plant biomass is lipopeptide. GCMS revealed that the crude oil (C7 – C23) was efficiently bio-degraded from lower to higher molecular weight. The application of natural surfactants in electokinetic remediation increased the plant biomass degradation of crude oil polluted soil by 98% compared to electrokinetic 55% in 2 days. Natural surfactant improves hydrocarbon solubilization and accelerates hydrocarbon electro migration to the anodic compartment, as confirmed by the presence of greater total organic content than the electrokinetic. This study proves that BIO-EK compared with a natural surfactant derived from plant biomass may be utilized to improve in situ bioremediation of crude oil polluted soils.