Hydrocarbon Biodegradation Rates Research Articles

Enhancing bioremediation of petroleum-contaminated soil by sophorolipids-modified biochar: Combined metagenomic and metabolomic analyses

In this study, sophorolipids (SLs)-modified biochar (BC-SLs) was used to enhance the bioremediation of petroleum hydrocarbons (PHs) contaminated soil. The biodegradation rate of petroleum hydrocarbons (PHs) by BC-SLs and BC treatments were 62.86 % and 52.64 % after 60 days of remediation experiments, respectively, higher than non-biochar treatment group (24.09 %). The metagenomic analysis showed that the abundance of petroleum-degrading bacteria Actinobacteria and Proteobacteria were increased by 3.8 % and 5.3 %, respectively in BC-SLs treatment, and the abundance of functional genes for PHs degradation, such as alkB, nidA and pcaG, were significantly increased by 12.85 %, 30.08 % and 21.01 %, respectively. The metabolomic analysis showed that BC-SLs facilitated the metabolic process of PHs, the microbial metabolism of petroleum hydrocarbons (PHs) became more active. Fatty acid degradation and polycyclic aromatic hydrocarbons (PAHs) degradation were up-regulated, indicating the promoting effect of the BC-SLs for PHs metabolism. The combined metagenomic and metabolomic analysis demonstrated the strong positive correlations between PHs metabolites and PHs-degrading bacteria, such as lauric acid vs. Actinobacteria, benzoic vs. Proteobacteria. The strong positive correlations between PHs metabolites and PHs-degrading genes were also observed, such as o-ehyltoluene vs. nahD, 4-isopropylbenzoic acid vs. etbAa. The modification of biochar with SLs increased the oxygen-containing functional groups on the surface of biochar. Meanwhile, the emulsification and solubilization of SLs promoted the bioavailability of PHs. The effects of BC-SLs on the nitrogen cycle during PHs remediation showed that it facilitated the accumulation of nitrogen-fixing genes, promoted nitrification but inhibited denitrification process. This study confirms that the application of BC-SLs is an effective remediation of PHs contamination and a sustainable method for controlling agricultural waste resources.

Characterization of hydrocarbon degraders from Northwest Passage beach sediments and assessment of their ability for bioremediation.

Global warming-induced sea ice loss in the Canadian Northwest Passage (NWP) will result in more shipping traffic, increasing the risk of oil spills. Microorganisms inhabiting NWP beach sediments may degrade hydrocarbons, offering a potential bioremediation strategy. In this study, the characterization and genomic analyses of 22 hydrocarbon-biodegradative bacterial isolates revealed that they contained a diverse range of key alkane and aromatic hydrocarbon-degradative genes, as well as cold and salt tolerance genes indicating they are highly adapted to the extreme Arctic environment. Some isolates successfully degraded Ultra Low Sulfur Fuel Oil (ULSFO) at temperatures as low as -5°C and high salinities (3%-10%). Three isolates were grown in liquid medium containing ULSFO as sole carbon source over 3 months and variation of hydrocarbon concentration was measured at three time points to determine their rate of hydrocarbon biodegradation. Our results demonstrate that two isolates (Rhodococcus sp. R1B_2T and Pseudarthrobacter sp. R2D_1T) possess complete degradation pathways and can grow on alkane and aromatic components of ULSFO under Arctic conditions. Overall, these results demonstrate that diverse hydrocarbon-degrading microorganisms exist in the NWP beach sediments, offering a potential bioremediation strategy in the events of a marine fuel spill reaching the shores of the NWP.

Metagenomic analysis revealed highly diverse carbon fixation microorganisms in a petroleum-hydrocarbon-contaminated aquifer

As one of the ultimate products of hydrocarbon biodegradation, inorganic carbon always be used to evaluate hydrocarbon biodegradation rates in petroleum-hydrocarbon-contaminated (PHC) aquifers. The evaluation method was challenged because of the existence of carbon fixation microorganisms, which may uptake inorganic carbons and consequently cause the biodegradation rates to be underestimated. We wonder if there are carbon fixation microorganisms in PHC aquifers. Although an extremely limited number of carbon fixation microorganisms in PHC sites have been studied in previous studies, the vast majority of microorganisms that participate in carbon fixation have not been systematically identified. To systematically reveal carbon fixation microorganisms and their survival environmental conditions, high-throughput metagenomic sequencing technologies, which are characterized by culture-independent, unbiased, and comprehensive methods for the detection and taxonomic characterization of microorganisms, were introduced to analyze the groundwater samples collected from a PHC aquifer. Results showed that 1041 genera were annotated as carbon fixation microorganisms, which accounted for 49% of the total number of genera in the PHC aquifer. Carbon fixation genes involved in Calvin-Benson-Bassham (CBB), 3-hydroxy propionate (3HP), reductive tricarboxylic acid (rTCA), and Wood-Ljungdahl (WL) cycles accounted for 2%, 41%, 34%, and 23% of the total carbon fixation genes, respectively, and 3HP, rTCA, and WL can be deemed as the dominant carbon fixation pathways. Most of the identified carbon fixation microorganisms are potential hydrocarbon biodegraders, and the most abundant carbon fixation microorganisms, such as Microbacterium, Novosphingobium, and Reyranella, were just the most abundant microorganisms in the aquifer system. It's deduced that most of the microorganisms in the aquifer were facultative autotrophic, and undertaking the dual responsibilities of degrading hydrocarbons to inorganic carbon and uptaking inorganic carbon to biomass.

Different strategies and bio-removal mechanisms of petroleum hydrocarbons from contaminated sites

PurposePetroleum hydrocarbons are naturally occurring flammable fossil fuels used as conventional energy sources. It has carcinogenic, mutagenic properties and is considered a hazardous pollutant. Soil contaminated with petroleum hydrocarbons adversely affects the properties of soil. This paper aim to remove pollutants from the environment is an urgent need of the hour to maintain the proper functioning of soil ecosystems.Design/methodology/approachThe ability of micro-organisms to degrade petroleum hydrocarbons makes it possible to use these microorganisms to clean the environment from petroleum pollution. For preparing this review, research papers and review articles related to petroleum hydrocarbons degradation by micro-organisms were collected from journals and various search engines.FindingsVarious physical and chemical methods are used for remediation of petroleum hydrocarbons contaminants. However, these methods have several disadvantages. This paper will discuss a novel understanding of petroleum hydrocarbons degradation and how micro-organisms help in petroleum-contaminated soil restoration. Bioremediation is recognized as the most environment-friendly technique for remediation. The research studies demonstrated that bacterial consortium have high biodegradation rate of petroleum hydrocarbons ranging from 83% to 89%.Social implicationsProper management of petroleum hydrocarbons pollutants from the environment is necessary because of their toxicity effects on human and environmental health.Originality/valueThis paper discussed novel mechanisms adopted by bacteria for biodegradation of petroleum hydrocarbons, aerobic and anaerobic biodegradation pathways, genes and enzymes involved in petroleum hydrocarbons biodegradation.

Effects of Dispersant on the Petroleum Hydrocarbon Biodegradation and Microbial Communities in Seawater from the Baltic Sea and Norwegian Sea.

Dispersants have been used in several oil spill accidents, but little information is available on their effectiveness in Baltic Sea conditions with low salinity and cold seawater. This study investigated the effects of dispersant use on petroleum hydrocarbon biodegradation rates and bacterial community structures. Microcosm experiments were conducted at 5 °C for 12 days with North Sea crude oil and dispersant Finasol 51 with open sea Gulf of Bothnia and coastal Gulf of Finland and Norwegian Sea seawater. Petroleum hydrocarbon concentrations were analysed with GC-FID. Bacterial community structures were studied using 16S rDNA gene amplicon sequencing, and the abundance of genes involved in hydrocarbon degradation with quantitative PCR. The highest oil degradation gene abundances and oil removal were observed in microcosms with coastal seawater from the Gulf of Bothnia and Gulf of Finland, respectively, and the lowest in the seawater from the Norwegian Sea. Dispersant usage caused apparent effects on bacterial communities in all treatments; however, the dispersant's effect on the biodegradation rate was unclear due to uncertainties with chemical analysis and variation in oil concentrations used in the experiments.

Quantification of biodegradation rate of hydrocarbons in a contaminated aquifer by CO2 δ13C monitoring at ground surface

Ground surface analysis of CO2 emissions with δ13C determination is experimentally demonstrated to be a potential methodology to monitor, on line, the dynamics of petroleum-hydrocarbon biodegradation in soil aquifers, thanks to the improvement of the Isotopic Ratio Infra Red Spectroscopy technique. Biodegradation rate of remaining hydrocarbon substrates in groundwater can be quantified using basic application of the Rayleigh equations, by δ13CCO2 analysis released at ground surface above the pollution plume instead of usual approaches based on groundwater hydrocarbons δ13C analysis, when physical and chemical properties for the contaminated site meet appropriate conditions.The validation approach for that gasoline contaminated specific site is discussed and verified by comparison of first order attenuation rate constant determined from δ13CCO2 analysis emitted at ground surface and from δ13CTOLUENE analysis in ground water. A kinetic fractionation factor α of 0.9979 (or ε value of −2.1 ± 0.5‰) is estimated for the biodegradation of the most reactive hydrocarbon substrates (TEX). The treatment of this Rayleigh equations by linear regression of δ13CCO2 values along the predominant direction of groundwater flow leads to the following results and conclusions for that site: (i) first order biodegradation rate constants (and annual variation) are maximum after the activation of a Permeable Reactive Barrier (PRB) in May 2014: 0.92(+0.29–0.17) year−1, and during July and October: 0.46(+0.14–0.09) year−1 and minimum in mid-winter in February 2015: 0.17(+0.05–0.03) year−1, given by the estimation range for ε. These results are in the lower range with reported in literature for similar contaminated sites (1.6–18 year−1) considering natural attenuation under sulfate reducing conditions and (ii) the seasonal variation of the first order biodegradation rate constant is mainly correlated with the seasonal variation of the CO2 flux, where maximum values are in summers and minimum values in winters. Both seasonal variations are mainly due to the annual cycle of the natural biodegradation activity at the scale of the pollution plume, rather than the activation of the PRB.This work demonstrates that δ13CCO2 analysis released at ground surface from biodegradation of groundwater hydrocarbons could provide, under characterized and appropriate conditions, a non-intrusive (without soil samplings), fast, and low-cost online method to monitor and therefore to optimize soil remediation processes in real time. (Monitored Natural Attenuation or Enhanced Bioremediation).

A Mathematical Model for Bioremediation of Hydrocarbon-Contaminated Soils

Bioremediation of hydrocarbons in soil is a highly complex process, involving a multiplicity of physical, chemical and biological phenomena. Therefore, it is extremely difficult to control and boost the bioremediation of these systems after an oil spill. A mathematical model was developed to assist in the prediction and decision-making regarding the in situ bioremediation of hydrocarbon-contaminated soils. The model considered the most relevant processes involved in the mass transfer and biodegradation of alkanes over time and along the depth of a flooded soil column. Aliphatic hydrocarbons were chosen since they are less water soluble than aromatics and account for 50–90% of the hydrocarbon fraction in several petroleum products. The effect of adding oxygen, nitrate, iron (III) or sulfate as electron acceptors was then simulated (bioremediation scenarios). Additionally, and to feed the model, batch assays were performed to obtain experimental data on hydrocarbon adsorption to soil particles (more than 60% of hydrocarbons tends to be adsorbed to soil particles), as well as hydrocarbon biodegradation rates in the presence of nitrate (0.114 d−1) and oxygen (0.587 d−1). The model indicates that saturated hydrocarbon removal occurs mainly with adsorption/desorption and transport processes in the upper layers of soil due to methanogenic biodegradation in deeper layers, since the other microbial processes are soon limited by the lack of electron acceptors. Simulation results show that higher initial electron acceptor concentrations led to higher hydrocarbon removal, confirming that the model is performing in accordance with the expected. Close to the surface (at 0.1 m depth), all scenarios predicted more than 83% hydrocarbon removal after two years of simulation. Soil re-aeration results in faster hydrocarbon removal (more than 20% after one year) and surfactants addition (around 15% after one year) may also accelerate soil bioremediation. With this model, the simultaneous contributions of the various physicochemical and biological processes are integrated, facilitating the simulation and comparison of different bioremediation scenarios. Therefore, it represents a useful support tool for the management of contaminated sites.

Emulating Deep-Sea Bioremediation: Oil Plume Degradation by Undisturbed Deep-Sea Microbial Communities Using a High-Pressure Sampling and Experimentation System

Hydrocarbon biodegradation rates in the deep-sea have been largely determined under atmospheric pressure, which may lead to non-representative results. In this work, we aim to study the response of deep-sea microbial communities of the Eastern Mediterranean Sea (EMS) to oil contamination at in situ environmental conditions and provide representative biodegradation rates. Seawater from a 600 to 1000 m depth was collected using a high-pressure (HP) sampling device equipped with a unidirectional check-valve, without depressurization upon retrieval. The sample was then passed into a HP-reactor via a piston pump without pressure disruption and used for a time-series oil biodegradation experiment at plume concentrations, with and without dispersant application, at 10 MPa and 14 °C. The experimental results demonstrated a high capacity of indigenous microbial communities in the deep EMS for alkane degradation regardless of dispersant application (>70%), while PAHs were highly degraded when oil was dispersed (>90%) and presented very low half-lives (19.4 to 2.2 days), compared to published data. To our knowledge, this is the first emulation study of deep-sea bioremediation using undisturbed deep-sea microbial communities.

THE PETROLEUM HYDROCARBON BIOREMEDIATION PERFORMANCE BY INOCULATING CONSORTIUM PETROPHYLIC AND THE CHICKEN MANURE AMENDMENT

Biodegradation of petroleum hydrocarbons in a land-treatment bioremediation system requires an effective degrading microbes and optimum system conditions. The study aimed to obtain the characteristics of the interaction between the petrophylic consortium (Pseudomonas sp. and Aspergillus sp.) and the amendment of chicken manure and to obtain the optimum dose to increase the biodegradation rate of petroleum hydrocarbons, soil pH change, and increased the population petrophylic fungi. The experiment was carried out on a greenhouse scale, using a factorial randomized block design (RBD) with two factors and three replications. The first factor was the inoculation of the petrophylic consortium consisting of: (i) without Petrophylic, (ii) 1% Petrophylic, (iii) 3% Petrophylic, (iv) 5% Petrophylic. The second factor was the amendment of chicken manure, consisting of 3 levels: (i) without chicken manure, (ii) 2% chicken manure, and (iii) 4% chicken manure. The results showed that petrophylic consortium inoculation and chicken manure amendment did not show an interaction effect on increasing the rate of biodegradation of petroleum hydrocarbons, soil pH, and increasing the population of petrophylic fungi. Likewise, the effect of independent treatment could not increase the biodegradation rate and soil pH.But, the application of chicken manure increased growth of the petrophylic fungus population during the 4-week incubation time. However, increasing Petrophylic fungi population density did not show increasing in degrading petroleum hydrocarbons. The dose of chicken manure used has not been able to supply N and P nutrients to achieve optimal C: N: P ratio conditions for degrading hydrocarbons effectively, resulting the inoculated petrophylic could not degrade hydrocarbons maximally. Compatibility between consortium microbes members as a hydrocarbon-degrading agent is an important factor for achieving effectively bioremediation performance.

Temporal variation of crude and refined oil biodegradation rates and microbial community composition in freshwater systems

Freshwater systems are vulnerable to contamination by oil extraction and transportation. Thus, it is critical to understand how large freshwater ecosystems such as the Great Lakes will respond to released oil. In this study, we investigated differences in the microbial response to oil in the Straits of Mackinac at different times throughout the year and if crude (Bakken) and refined (non-highway diesel) oil exposure differentially altered the microbial community composition and hydrocarbon biodegradation rates. We also investigated the impact of temperature on the microbial response to oil by incubating samples collected in October of 2018 at 23 °C and at 4 °C. Ambient microbial communities differed between sample collection times, with significantly enriched microbial groups present between most sample types. We found significantly different microbial communities between control and oil-amended samples, but no significant differences between either oil type. We found that the bacterial family Solimonadaceae were significantly enriched in all oil-amended microcosms compared to the control microcosms across sampling times. We assessed oil biodegradation using CO2 production as a proxy for hydrocarbon metabolism. We observed a general trend of increased respiration rates with oil amendment compared to the control. No statistically significant differences in daily CO2 production rates existed between the two oil types. These findings suggest that microbial community in the Straits of Mackinac shifts over time even without oil amendment, and that the microbial communities in the Straits of Mackinac are compositionally and metabolically responsive to the presence of varying oil types throughout the year.

Effects of Dispersants and Biosurfactants on Crude-Oil Biodegradation and Bacterial Community Succession.

This study evaluated the effects of three commercial dispersants (Finasol OSR 52, Slickgone NS, Superdispersant 25) and three biosurfactants (rhamnolipid, trehalolipid, sophorolipid) in crude-oil seawater microcosms. We analysed the crucial early bacterial response (1 and 3 days). In contrast, most analyses miss this key period and instead focus on later time points after oil and dispersant addition. By focusing on the early stage, we show that dispersants and biosurfactants, which reduce the interfacial surface tension of oil and water, significantly increase the abundance of hydrocarbon-degrading bacteria, and the rate of hydrocarbon biodegradation, within 24 h. A succession of obligate hydrocarbonoclastic bacteria (OHCB), driven by metabolite niche partitioning, is demonstrated. Importantly, this succession has revealed how the OHCB Oleispira, hitherto considered to be a psychrophile, can dominate in the early stages of oil-spill response (1 and 3 days), outcompeting all other OHCB, at the relatively high temperature of 16 °C. Additionally, we demonstrate how some dispersants or biosurfactants can select for specific bacterial genera, especially the biosurfactant rhamnolipid, which appears to provide an advantageous compatibility with Pseudomonas, a genus in which some species synthesize rhamnolipid in the presence of hydrocarbons.

Quantifying natural source zone depletion at petroleum hydrocarbon contaminated sites: A comparison of 14C methods

Surficial CO2 efflux surveys have been used to delineate hydrocarbon source zones in contaminated aquifers and provide estimates of hydrocarbon biodegradation rates. This approach requires distinguishing between CO2 derived from petroleum degradation and CO2 produced from natural soil respiration. To this end, radiocarbon has been used to differentiate between 14C–depleted CO2 from hydrocarbon degradation and 14C–enriched CO2 from natural soil respiration to effectively quantify the contribution of each source to total CO2 efflux, and by deduction natural source zone depletion (NSZD) rates. In this study, a systematic method comparison has been conducted to evaluate available approaches for collecting CO2 gas samples for radiocarbon analysis used to correct total CO2 efflux measurements for quantifying natural source zone depletion rates. Gas samples for radiocarbon analysis were sampled from (i) dynamic closed chambers (located at ground surface), (ii) static chambers (also at ground surface), (iii) shallow soil gas probes (0.3 m bgs), and (iv) soil gas monitoring wells (~0.6 m below ground surface) during a CO2 efflux survey conducted at the site of a historical pipeline rupture near Bemidji, MN. The mean fraction of radiocarbon (F14C) obtained from samples overlying the source zone were (i) 0.93 ± 0.01, (ii) 0.73 ± 0.03, (iii) 0.71 ± 0.04, and (iv) 0.41 ± 0.06, for the four methods respectively. These F14C values were used to apportion total CO2 efflux measurements into contributions of contaminant–derived CO2 efflux and natural soil respiration to evaluate natural source zone depletion processes. Results suggest that the method of radiocarbon sampling has a significant effect on the calculated fraction of the CO2 efflux originating from contaminant-related soil respiration, with contributions ranging between 27% and 59% of total soil respiration. Results indicate that radiocarbon sampled from static chambers and shallow soil gas probes methods offer the best compromise between CO2 sample yield and sample representativeness, providing the most reliable estimates of CO2 effluxes originating from contaminant degradation. However, the results also show that for this study, all methods agree within a factor of <2.3 regarding the inferred NSZD rates.

Initial oil concentration affects hydrocarbon biodegradation rates and bacterial community composition in seawater

During oil spills in the field or for laboratory incubation studies, different oil concentrations are often encountered or applied, yet how initial oil concentration affects biodegradation rates of hydrocarbons and the development of oil degraders remains unclear. We incubated seawater for 50 d with different oil concentrations (0, 50, 100, 200, 400 and 800 ppm). n-Alkanes and polycyclic aromatic hydrocarbons (PAHs), and the bacterial community were analyzed periodically. Results show that the biodegradation rates of alkanes, derived from first order kinetics, decreased with increasing oil concentration, but percent residual was ~50% regardless of the initial concentration. In contrast, the biodegradation rates of PAHs increased with concentration, and the percent residual increased with oil concentration. Increasing oil concentration resulted in increased abundances of Rhodobacterales, Altererythrobacter, and Neptuniibacter. However, Alcanivorax abundance was barely detected in 400 and 800 ppm. Overall, oil concentration critically affected the degradation of hydrocarbons and the bacterial community.

Potensi Tanaman Sorgum (Sorghum bicolor L.), Azotobacter sp. dan Pseudomonas sp. Sebagai agen biologis dalam proses Fitoremediasi Hidrokarbon minyak bumi

Phytoremediation using Sorghum (Sorghum bicolor L.) plant is an alternative green technology to overcome oil polluted soil. Improving of the phytoremediation performance are needed stimulating agent to enhance the rate of hydrocarbon biodegradation and increase the plant growth. One of the bioagent that can act as biostimulant is Azotobacter sp. and Pseudomonas sp., because thouse bioagent can provide elements N and P, and phytohormone for sorghum plants. The purpose of this study is to examine the role of Azotobacter sp. and Pseudomonas sp. as a biostimulant for Sorghum bicolor L. in the process of biodegradation of petroleum hydrocarbon in soil system. The factorial randomized block design was used as the experimental design. The treatment consisted of 4 application levels of Azotobacter sp. and 4 levels of application of Pseudomonas sp. The results showed that there was no interaction between Azotobacter sp. and Pseudomonas sp. aplication on the efficiency of hydrocarbon degradation of the population of Azotobacrer spp., Pseudomonas spp. and plant height increase. Azotobacter sp. and Pseudomonas sp. also did not show an increase in value on each test variable. While, the potential of sorghum plants without the application of Azotobacter sp. and Pseudomonas sp. (control treatment) showed the value of the efficiency of biodegradation in the range of values of 60.442% - 68.165% during 14 weeks period and not significantly different from other treatments.

Enhanced bioremediation of diesel range hydrocarbons in soil using biochar made from organic wastes.

Hydrocarbon contamination due to anthropogenic activities is a major environmental concern worldwide. The present study focuses on biochar prepared from fruit and vegetable waste and sewage sludge using a thermochemical approach and its application for the enhanced bioremediation (biostimulation and bioaugmentation) of diesel-polluted soil. The biochar was characterized using FTIR (Fourier-transform infrared spectroscopy), elemental analysis, surface area analysis, and pore analysis. Adsorption experiments showed that hydrocarbon degradation was attributed to biological processes rather than adsorption. The study found that various biochar amendments could significantly increase the rate of hydrocarbon biodegradation with removal efficiencies > 70%. Bioaugmentation using cow dung further improved the removal efficiency to 82%. Treatments showing the highest degree of removal efficiency indicated the presence of 27 different bacteria phyla with Proteobacteria and Actinobacteria as the most abundant phyla. The present study concludes that biochar amendments have great potential for enhancing the bioremediation of soils contaminated with diesel range hydrocarbons.

Surfactin for enhanced removal of aromatic hydrocarbons during biodegradation of crude oil

Oil biodegradation studies using the powerful surfactant have mainly focused on saturated hydrocarbons or parent aromatic hydrocarbons, but not specifically on the position or degree of alkyl substitution of aromatic hydrocarbons. Removal of substituted aromatic hydrocarbons remains a controversial topic in biodegradation. In this study, the effect of surfactin addition on specific alkylated aromatic isomers and saturated hydrocarbons was observed in aerobic biodegradation experiments with four species of bacteria. The relative susceptibility of the individual alkylphenanthrene, alkyldibenzothiophene and methyltriaromatic steroid isomers to biodegradation was determined by their depletion rates. The results show that different bacteria induce removal of alkylated aromatic hydrocarbons and alteration of other petroleum hydrocarbons in various ways. Bacillus amyloliquefaciens strain BC is the best for biodegradation of general petroleum hydrocarbons, because it can convert n-alkanes, n-alkylcyclohexanes, isoprenoids, and aromatic hydrocarbons. Achromobacter strain J3 can degrade methylphenanthrenes, methyltriaromatic steroids, triaromatic steroids, and steranes better than other bacteria. Citrobacter sp strain J1 induces the highest degradation rate for dimethylphenanthrenes, trimethylphenanthrenes, and dimethyldibenzothiophenes, and Brucella melitensis strain J2 selectively degrades methyldibenzothiophenes. Surfactin addition increases the biodegradation rate of alkylaromatic hydrocarbons by 1.5–87.2%, and can significantly enhance the biodegradation of alkylphenanthrenes, alkyldibenzothiophenes and methyltriaromatic steroids. Alkyldibenzothiophenes can be used as markers for determining the levels of biodegradation of crude oils, and when used in conjunction with triaromatic steroids are a powerful indicator of the biodegradation of petroleum. The use of surfactin for enhancing the removal of aromatic hydrocarbons provides a wide range of applications in the environmental remediation and petroleum industry.

Synergistic Effect of Rhamnolipids and Inoculation on the Bioremediation of Petroleum-Contaminated Soils by Bacterial Consortia.

Crude oil is a serious soil pollutant, requiring large-scale remediation efforts. Bacterial consortia in combination with rhamnolipids can be an effective bioremediation method. However, the underlying mechanisms and associated changes in soil bacterial composition remain uncharacterized. Therefore, this study sought to evaluate the effectiveness of rhamnolipids in petroleum hydrocarbon removal, and the associated bacterial community dynamics during bioremediation of petroleum-contaminated soils. Contaminated soils were subjected to natural attenuation, bioremediation with rhamnolipids, bioremediation with bacterial consortia, or bioremediation with bacterial consortia supplemented with rhamnolipids (BMR). High-throughput sequencing of bacterial sample partial 16S rRNA sequences was performed. Additionally, the n-alkanes and aromatic fractions were analyzed by gas chromatography-mass spectroscopy. The results showed that rhamnolipid supplementation increased the rate and extent of total petroleum hydrocarbon biodegradation to a maximum of 81% within 35days. Further, phylogenetic analysis revealed that the bacterial community was composed of 14 phylotypes (similarity level = 97%). Actinobacteria and Proteobacteria were the two core phyla in all samples, accounting for 63-89%, but Proteobacteria was the most dominant phylum in the BMR sample (~ 53%). Among the top 20 genera, Pseudomonas, Pseudoxanthomonas, Cavicella, Mycobacterium, Rhizobium, and Acinetobacter were more abundant in BMR samples compared to other samples. Predicted functional profiles revealed that rhamnolipid addition also induced changes in gene abundance related to hydrocarbon metabolic pathways. This study provided comprehensive insights into the synergistic effect of rhamnolipids and bacterial consortia for altering bacterial populations and specific functional traits, which may serve to improve bacteria-mediated petroleum hydrocarbon biodegradation in contaminated soils.

Dispersant Enhances Hydrocarbon Degradation and Alters the Structure of Metabolically Active Microbial Communities in Shallow Seawater From the Northeastern Gulf of Mexico.

Dispersant application is a primary emergency oil spill response strategy and yet the efficacy and unintended consequences of this approach in marine ecosystems remain controversial. To address these uncertainties, ex situ incubations were conducted to quantify the impact of dispersant on petroleum hydrocarbon (PHC) biodegradation rates and microbial community structure at as close as realistically possible to approximated in situ conditions [2 ppm v/v oil with or without dispersant, at a dispersant to oil ratio (DOR) of 1:15] in surface seawater. Biodegradation rates were not substantially affected by dispersant application at low mixing conditions, while under completely dispersed conditions, biodegradation was substantially enhanced, decreasing the overall half-life of total PHC compounds from 15.4 to 8.8 days. While microbial respiration and growth were not substantially altered by dispersant treatment, RNA analysis revealed that dispersant application resulted in pronounced changes to the composition of metabolically active microbial communities, and the abundance of nitrogen-fixing prokaryotes, as determined by qPCR of nitrogenase (nifH) genes, showed a large increase. While the Gammaproteobacteria were enriched in all treatments, the Betaproteobacteria and different families of Alphaproteobacteria predominated in the oil and dispersant treatment, respectively. Results show that mixing conditions regulate the efficacy of dispersant application in an oil slick, and the quantitative increase in the nitrogen-fixing microbial community indicates a selection pressure for nitrogen fixation in response to a readily biodegradable, nitrogen-poor substrate.

EVALUATION OF DIFFERENT SCREENING METHODS FOR BIOSURFACTANT PRODUCERS ISOLATED FROM EGYPTIAN FRESH WATER SAMPLES CONTAMINATED BY OIL SPILLS USING BACILLUS SUBTILIS AND BACILLUS LICHENIFORMIS

In all countries, oil exploration and use threatens the health of the environment and living creatures including humans. An oil spill is the release of petroleum hydrocarbon into the environment. One of the most applicable and safe method is the bioremediation treatment, using microorganisms .This work aims at treating oil spills in fresh water and to compile information on types and properties of biosurfactant. It also describes factors affecting these biosufactants production. Different screening methods e.g. oil spreading assay, Emulsification index (E24), Drop collapse assay, Blood agar test, Hemolysis, MaCconkey agar test were tested. Two isolates gave highly Emulsification tests named O3, O11, which were identified as Bacillus subtilis and Bacillus licheniformis respectively, were isolated from petroleum hydrocarbon contaminated water. Optimization of two isolates were done and highest biosurfactant activity at temperature ranges from 10 oC to 50 oC and pH ranges from 3 to 10 and salinity ranges from 0%to 30%. Also, the biosurfactant was characterized by Fourier transform infrared spectroscopy (FTIR) and surface tension measurement. FTIR showed the production of biosurfactant from Bacillus subtilis similar to surfactin, while the biosurfactant from Bacillus licheniformis similar to lichenysins. The use of microbial biosurfactants significantly decreases the hydrophobicity and increases the rate of hydrocarbon biodegradation. Biosurfactants obtain demonstrated good surface tension reduction capacity by Bacillus subtilis and Bacillus licheniformis up to 30 and 36Nm/m and have emulsifying activity 71% and 65% respectively.

Effect of spatial origin and hydrocarbon composition on bacterial consortia community structure and hydrocarbon biodegradation rates.

Oil reserves in deep-sea sediments are currently subject to intense exploration, with associated risks of oil spills. Previous research suggests that microbial communities from deep-sea sediment (>1000m) can degrade hydrocarbons (HCs), but have a lower degradation ability than shallow (<200m) communities, probably due to in situ temperature. This study aimed to assess the effect of marine origin on microbial HC degradation potential while separating the influence of temperature, and to characterise associated HC-degrading bacterial communities. Microbial communities from 135 and 1000 m deep sediments were selectively enriched on crude oil at in situ temperatures and both consortia were subsequently incubated for 42 days at 20°C with two HC mixtures: diesel fuel or model oil. Significant HC biodegradation occurred rapidly in the presence of both consortia, especially of low molecular weight HCs and was concomitant with microbial community changes. Further, oil degradation was higher with the shallow consortium than with the deep one. Dominant HC-degrading bacteria differed based on both spatial origin of the consortia and supplemented HC types. This study provides evidence for influence of sediment spatial origin and HC composition on the selection and activity of marine HC-degrading bacterial communities and is relevant for future bioremediationdevelopments.