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.

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.

Increasing anthropogenic activities in the Arctic represent an enhanced threat for oil pollution in a marine environment that is already at risk from climate warming. In particular, this applies to species with free-living pelagic larvae that aggregate in surface waters and under the sea ice where hydrocarbons are likely to remain for extended periods of time due to low temperatures. We exposed the positively buoyant eggs of polar cod (Boreogadus saida), an arctic keystone species, to realistic concentrations of a crude oil water-soluble fraction (WSF), mimicking exposure of eggs aggregating under the ice to oil WSF leaking from brine channels following encapsulation in ice. Total hydrocarbon and polycyclic aromatic hydrocarbon levels were in the ng/L range, with most exposure concentrations below the limits of detection throughout the experiment for all treatments. The proportion of viable, free-swimming larvae decreased significantly with dose and showed increases in the incidence and severity of spine curvature, yolk sac alterations and a reduction in spine length. These effects are expected to compromise the motility, feeding capacity, and predator avoidance during critical early life stages for this important species. Our results imply that the viability and fitness of polar cod early life stages is significantly reduced when exposed to extremely low and environmentally realistic levels of aqueous hydrocarbons, which may have important implications for arctic food web dynamics and ecosystem functioning.

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.

Crude oil is a complex mixture of petroleum hydrocarbons (PHC) with distinct chemical, physical and toxicological properties relevant for contaminated site risk assessment. Ecotoxicological effects of crude oil distillation fractions on luminescent bacteria (Vibrio fischeri), earthworms (Dendrobaena hortensis) and invertebrates (Heterocypris incongruens) were tested using two spiked soils and their elutriates. Fraction 2 (F2) had an equivalent carbon number (ECN) range of >10 to 16, and F3 from >16 to 39. F2 showed a substantially higher ecotoxicological effect than F3 for Vibrio and Dendrobaena. In contrast, severe inhibition of Heterocypris by the poorly soluble F3 is attributed to mechanical organ blockage. Immediate sequestration of PHC to the organic matter-rich soil effected reduced toxicity for all organisms. This study indicates that a more differentiated consideration (i) of PHC mixtures based on ECN range and (ii) of model soil properties employed for ecotoxicity testing should be included into PHC-contaminated site risk assessment.

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.

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.

A pyrene-degrading consortium OPK containing Mycolicibacterium strains PO1 and PO2, Novosphingobium pentaromativorans PY1 and Bacillus subtilis FW1 effectively biodegraded medium- and long-chain alkanes as well as mixed hydrocarbons in crude oil. The detection of alkB and CYP153 genes in the genome of OPK members supports its phenotypic ability to effectively degrade a broad range of saturated hydrocarbons in crude oil. Zeolite-immobilized OPK was developed as a ready-to-use bioproduct and it exhibited 74% removal of 1000 mg L⁻¹ crude oil within 96 h in sterilized seawater without nutrient supplementation and maintained high crude oil-removal activity under a broad range of pH values (5.0–9.0), temperatures (30–40 °C) and salinities (20–60‰). In addition, the immobilized OPK retained a high crude oil removal efficacy in semicontinuous experiments and showed reusability for at least 5 cycles. Remarkably, bioaugmentation with zeolite-immobilized OPK in sandy soil microcosms significantly increased crude oil (10,000 mg kg⁻¹ soil) removal from 45% to 80.67% within 21 days compared to biostimulation and natural attenuation. Moreover, bioaugmentation with exogenous immobilized OPK stimulated an increase in the relative abundances of Alcanivorax genus, indigenous hydrocarbon-degrading bacteria, which in turn enhanced removal efficiency of crude oil contamination from sandy soil microcosms. The results indicate positive interactions between the bioaugmented immobilized consortium, harboring Mycolicibacterium as a key player, and indigenous Alcanivorax, which exhibited crucial functions for improving crude oil removal efficacy. The knowledge obtained forms an important basis for further synthesis and handling of a promising bio-based product for enhancing the in situ bioremediation of crude oil-polluted marine environments.