Naphthenic acids are a group of polar organic carboxylic acids that are present in crude oil naturally. They are cycloaliphatic carboxylic acids which have 10 to 16 carbons, which gained importance since the early twentieth century because of corrosion in oil refineries. Moreover, they are the most important environmental pollutants caused by oil extraction from oil sand reserves. Heavy crude oils which have high concentration of naphthenic acids are usually considered as poor-quality oil and sold at a lower price. Often, the high concentration of naphthenic acids in crude oil reduces the life of the equipment which are used in the exploration and refining process because of corrosion. Hence, researchers are increasingly interested in the chemical properties of naphthenic acids and the acidic components of the crude oils. The most popular methods for the identification and analysis of naphthenic acids are liquid and gas chromatography (GC), liquid-liquid extraction, Fourier transform infrared spectroscopy (FTIR), and solid-phase extraction (SPE). Naphthenic acids are the most important environmental pollutants caused by oil extraction from oil sand reserves. Previous studies have revealed that naphthenic acids can be absorbed by fish, but their distribution in different tissues of fish has not been specified. Experimental samples showed the highest toxicity to fish, while there was less toxicity to invertebrates and algae. Moreover, naphthenates have various industrial utilizations; they are used in synthetic detergents, corrosion inhibitors, lubricants, fuel and oil additives, wood preservatives, insecticides, fungicides, pesticides, wetting agents, napalm thickening agents, and oil desiccants that are utilized in painting and treating wood surfaces.

Middle and southern Iraq suffers from polluted soils due to crude oil, spilled on land, leakage from transmitting pipe networks, or petroleum products from refineries. Many researchers have studied pollution impacts on the soil in details, but there is a clear lack of investigation on the influence of crude oil on soil erodibility. Recent researches have investigated the influence of pollution on erodibility parameters, which include critical shear stress (τc) and detachment factor (dc). The variability of dc and τc due to different in-situ scaling has not been thoroughly established for polluted and unpolluted soils. Thus this research aims at investigating the influence of different in-situ scaling ratios (1:1, 1:30, and 1:50) on variability of dc and τc for polluted and unpolluted soils under controlled laboratory conditions, using Jet Erosion Test (JET), and tries to compare the three solution techniques (namely, Blaisdell’s approach, depth scour approach, and iterative approach) to solve dc and τc from JETs for polluted and unpolluted soils. The polluted soil samples have been prepared by submerging the soil surface with crude oil for 24 hours prior to testing. Results show that there have been statistical differences in dc and τc between polluted and unpolluted soil samples on the dry side of water contents with no statistically significant difference of measured dc and τc being observed across different in-situ scale ratios for polluted and unpolluted soils. All told, the study shows less variability of measured dc and τc across different solution techniques, compared to previous study findings.

As ice extent in the Arctic is declining, oil and gas activities will increase, with higher risk of oil spills to the marine environment. To determine biotransformation of dispersed weathered oil in newly formed ice, oil dispersions (2–3 ppm) were incubated in a mixture of natural seawater and frazil ice for 125 days at −2 °C. Dispersed oil in seawater without frazil ice were included in the experimental setup. Presence or absence of frazil ice was a strong driver for microbial community structures and affected the rate of oil degradation. n-alkanes were degraded faster in the presence of frazil ice, the opposite was the case for naphthalenes and 2–3 ring PAHs. No degradation of 4–6 ring PAHs was observed in any of the treatments. The total petroleum oil was not degraded to any significant degree, suggesting that oil will freeze into the ice matrix and persist throughout the icy season. | publishedVersion

The interaction of dispersed oil droplets with large diameter suspended particulate materials (SPM) has been little studied. In the current study, particle size, oil characteristics and chemical dispersant significantly influence the adsorption of oil droplets to SPM in seawater. Sediments with a smaller particulate size (clay) approaching that of the oil droplets (2–20 lm) adsorbed more oil per gram than sediments with large particle size (sand). Heavier, more polar oils with a high asphaltene content adsorbed more efficiently to SPM than lighter, less polar oils. A decrease in the smaller, more water soluble oil components in the sediment adsorbed oil was observed for all oil types. Addition of chemical dispersant decreased the adsorption of oil droplets to suspended carbonate sand in an exponential-like manner. No change in the relative distribution of compounds adsorbed to the sediment was observed, indicating dispersants do not alter the dissolution of compounds from oil droplets. | acceptedVersion

Knowledge of the transport and fate of pathogenic Escherichia coli, especially in the areas contaminated with crude oil, is required to assess contamination of shallow groundwater resources. The present study aims to investigate the effect of crude oil-mediated water repellency on the movement of nalidixic acid-resistant Escherichia coli strain (E. coli NAR) and bromide (Br) as an inert tracer in two soil types. The soils were contaminated at three levels of 0, 0.5 and 1% w/w of total petroleum hydrocarbons (TPHs) using crude oil. Steady-state saturated flow in the soil columns was controlled using a tension infiltrometer. Leaching experiments were conducted through the columns of repacked (un-weathered) and physically-weathered clay loam (CL) and sandy loam (SL) soils. The columns leachate was sampled at specific times for 4 pore volumes. The shape of breakthrough curves for the E. coli NAR and Br depended on soil texture and structure and the TPHs level. Preferential flow in the crude oil-mediated water-repellent soils facilitated the transport of contaminants especially E. coli NAR. Filtration coefficient and relative adsorption index of bacteria were greatest in the repacked CL soils and were lowest in the weathered SL soils. Discontinuity of soil pores and lower flow velocity resulted in greater filtration of E. coli NAR in the repacked CL soil than other treatments. Physical weathering induced the formation of aggregates which reduced soil particle surfaces available for retention of water-repellent oil and contaminants. Movement of both bacteria and Br tracer in the weathered SL soil with 1% TPHs was higher than other treatments. This finding was attributed to low specific surface area, continuity of the pores and water repellency-mediated preferential pathways in the weathered SL soil columns. Our findings implied that shallow groundwater resources could be very sensitive to microbial contamination particularly in the oil-mediated water-repellent soils.

Crude oil contaminant is one of the major problem to environment and its removal process considered as most challenging tool currently across the world. In this degradation study, crude oil hydrocarbons are degraded on various pH optimization conditions (pH 2, 4,6,7,8 and 10) by using two biosurfactant producing bacterial strains Pseudomonas aeruginosa PP3 and Pseudomonas aeruginosa PP4. During crude oil biodegradation, degradative enzymes alkane hydroxylase and alcohol dehydrogenase were examined and found to be higher in PP4 than PP3. Biodegradation efficiency (BE) of crude oil by both PP3 and PP4 were analysed by gas chromatography mass spectroscopy (GCMS). Based on strain PP3, the highest BE was observed in pH 2 and pH 4 were found to be 62% and 69% than pH 6, 7, 8 and 10 (47%, 47%, 49% and 45%). It reveals that PP3 was survived effectively in acidic condition and utilized the crude oil hydrocarbons. In contrast, the highest BE of PP4 was observed in pH 7 (78%) than pH4 (68%) and pH's 2, 6, 8 and 10 (52%, 52%, 43% and 53%) respectively. FTIR spectra results revealed that the presence of different functional group of hydrocarbons (OH, –CH₃, CO, C–H) in crude oil. GCMS results confirmed that both strains PP3 and PP4 were survived in acidic condition and utilized the crude oil hydrocarbons as sole carbon sources. This is the first observation on biodegradation of crude oil by the novel strains of Pseudomonas aeruginosa in acidic condition with higher BE. Overall, the extracellular enzymes and surface active compounds (biosurfactant) produced by bacterial strains were played a key role in crude oil biodegradation process.

In the present study, produced water sample collected from the Indian crude oil reservoir is used to enrich the bacterial communities. The impact of these enriched bacterial communities on the biodegradation of crude oil, biofilm formation, and biocorrosion process are elucidated. A crude oil degradation study is carried out with the minimal salt medium and 94% of crude oil was utilized by enriched bacterial communities. During the crude oil degradation many enzymes including alkane hydroxylase, alcohol dehydrogenase, and lipase are playing a key role in the biodegradation processes. The role of enriched bacterial biofilm on biocorrosion reactions are monitored by weight loss studies and electrochemical analysis. Weight loss study revealed that the biotic system has vigorous corrosion attacks compared to the abiotic system. Both AC-Impedance and Tafel analysis confirmed that the nature of the corrosion reaction take place in the biotic system. Very less charge transfer resistance and higher corrosion current are observed in the biotic system than in the abiotic system. Scanning electron microscope confirms that the dense biofilm formation favoured the pitting type of corrosion. X-ray diffraction analysis confirms that the metal oxides formed in the corrosion systems (biotic). From the metagenomic analysis of the V3–V4 region revealed that presence of diverse bacterial communities in the biofilm, and most of them are uncultured/unknown. Among the known genus, Bacillus, Halomonas, etc are dominant in the enriched bacterial biofilm sample. From this study, we conclude that the uncultured bacterial strains are found to be playing a key role in the pitting type of corrosion and they can utilize crude oil hydrocarbons, which make them succeeded in extreme oil reservoir environments.

Fungi are well associated with the degradation of hydrocarbons by the production of different enzymes, among which catalases (CBH), laccases (LCC) and peroxidases (LiP and MnP) are of immense importance. In this study, crude oil tolerance and enzyme secretions were demonstrated by rhizospheric fungal strains. Four most abundant strains were isolated from the rhizosphere of grasses growing in aged oil spill sites and identified through morphological characterization and molecular PCR-amplification of 5.8–28S ribosomal rRNA using ITS1 and ITS4 primers. These strains were subjected to crude oil tolerance test at 0–20% concentrations. Presence and transcriptase responses of putative genes lig (1–6), mnp, cbh (1.1, 1.1 and 11), and lcc encoding lignin peroxidase, manganese peroxidase, catalase, and laccase enzymes respectively were also studied in these strains using RT-PCR. In addition, activities of secreted enzymes by each strain were studied in aliquots. The strains were identified as Aspergillus niger asemoA (KY473958), Talaromyces purpurogenus asemoF (KY488463), Trichoderma harzianum asemoJ (KY488466), and Aspergillus flavus asemoM (KY488467) through sequencing and comparing the sequences’ data at NCBI BLAST search software. All the isolated strains showed tolerance to crude oil at 20% concentration, but the growth rate reduced with increasing in oil concentrations. All the isolated strains possess the tested genes and lig 1–6 gene was overexpressed in A. niger and T. harzianum while lcc and mnp genes were moderately expressed in all the four strains. Almost 145 U.mL⁻¹ of lignin and manganese peroxidase, 87 U.mL⁻¹ of catalase, and 180 U.mL⁻¹ of laccase enzymes were produced by these strains and it was also observed that these strain mostly produced studied enzymes in response to increasing crude oil concentrations. Considering the robust nature and diverse production of these catalytic enzymes by these strains, they can be exploited for various bioremediation technologies as well as other biotechnological applications.

Bacterial degradation of crude oil in response to nutrient treatments has been vastly studied. But there is a paucity of information on kinetic parameters of crude oil degradation. Here we report the nutrient stimulated kinetic parameters of crude oil degradation assessed in terms of CO2 production and oil removal by Pseudomonas aeruginosa AKS1 and Bacillus sp. AKS2. The hydrocarbon degradation rate of P. aeruginosa AKS1 in oil only amended sediment was 10.75 ± 0.65 μg CO2-C g−1 sediment day−1 which was similar to degradation rate in sediments with no oil. In presence of both inorganic N & P, the degradation rate increased to 47.22 ± 1.32 μg CO2-C g−1 sediment day−1. The half-saturation constant (Ks) and maximum degradation rate (Vmax) for P. aeruginosa AKS1 under increasing N and saturating P concentration were 13.57 ± 0.53 μg N g−1 sediment and 39.36 ± 1.42 μg CO2-C g−1 sediment day−1 respectively. The corresponding values at increasing P and a constant N concentration were 1.60 ± 0.13 μg P g−1 sediment and 43.90 ± 1.03 μg CO2-C g−1 sediment day−1 respectively. Similarly the degradation rate of Bacillus sp. AKS2 in sediments amended with both inorganic nutrients N & P was seven fold higher than the rates in oil only or nutrient only treated sediments. The Ks and Vmax estimates of Bacillus sp. AKS2 under increasing N and saturating P concentration were 9.96 ± 1.25 μg N g−1 sediment and 59.96 ± 7.56 μg CO2-C g−1 sediment day−1 respectively. The corresponding values for P at saturating N concentration were 0.46 ± 0.24 μg P g−1 sediment and 63.63 ± 3.54 μg CO2-C g−1 sediment day−1 respectively. The rates of CO2 production by both isolates were further stimulated when oil concentration was increased above 12.5 mg g−1 sediment. However, oil degradation activity declined at oil concentration above 40 mg g−1 sediment when treated with constant nutrient: oil ratio. Both isolates exhibited alkane hydroxylase activity but aromatic degrading catechol 1, 2-dioxygenase and catechol 2, 3-dioxygenase activities were shown by P. aeruginosa AKS1 only.

We investigated how additions of choline, a naturally occurring methylated nitrogen-containing compound, accelerated hydrocarbon degradation in sandy sediments contaminated with moderately weathered crude oil (4000 mg kg−1 sediment). Addition of lauroylcholine chloride (LCC) and tricholine citrate (TCC) to oil contaminated sediments resulted in 1.6 times higher hydrocarbon degradation rates compared to treatments without added choline derivatives. However, the degradation rate constant for the oil contaminated sediments amended with LCC was similar to that in contaminated sediments amended with inorganic nitrogen, phosphorus, and glucose. Additions of LLC and TCC to sediments containing extensively weathered oil also resulted in enhanced mineralization rates. Cultivation-free 16S rRNA analysis revealed the presence of an extant microbial community with clones closely related to known hydrocarbon degraders from the Gammaproteobacteria, Alphaproteobacteria, and Firmicutes phyla. The results demonstrate that the addition of minimal amounts of organic compounds to oil contaminated sediments enhances the degradation of hydrocarbons.