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The environmental and economic impact of an oil spill can be significant. Biotechnologies applied during a marine oil spill involve bioaugmentation with immobilised or encapsulated indigenous hydrocarbonoclastic species selected under laboratory conditions to improve degradation rates. The environmental factors that act as stressors and impact the effectiveness of hydrocarbon removal are one of the challenges associated with these applications. Understanding how native microbes react to environmental stresses is necessary for effective bioaugmentation. Herein, Micrococcus luteus and M. yunnanensis isolated from a marine oil spill mooring system showed hydrocarbonoclastic activity on Maya crude oil in a short time by means of total petroleum hydrocarbons (TPH) at 144 h: M. luteus up to 98.79 % and M. yunnanensis 97.77 % removal. The assessment of Micrococcus biofilms at different temperature (30 °C and 50 °C), pH (5, 6, 7, 8, 9), salinity (30, 50, 60, 70, 80 g/L), and crude oil concentration (1, 5, 15, 25, 35 %) showed different response to the stressors depending on the strain. According to response surface analysis, the main effect was temperature > salinity > hydrocarbon concentration. The hydrocarbonoclastic biofilm architecture was characterised using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Subtle but significant differences were observed: pili in M. luteus by SEM and the topographical differences measured by AFM Power Spectral Density (PSD) analysis, roughness was higher in M. luteus than in M. yunnanensis. In all three domains of life, the Universal Stress Protein (Usp) is crucial for stress adaptation. Herein, the uspA gene expression was analysed in Micrococcus biofilm under environmental stressors. The uspA expression increased up to 2.5-fold in M. luteus biofilms at 30 °C, and 1.3-fold at 50 °C. The highest uspA expression was recorded in M. yunnanensis biofilms at 50 °C with 2.5 and 3-fold with salinities of 50, 60, and 80 g/L at hydrocarbon concentrations of 15, 25, and 35 %. M. yunnanensis biofilms showed greater resilience than M. luteus biofilms when exposed to harsh environmental stressors. M. yunnanensis biofilms were thicker than M. luteus biofilms. Both biofilm responses to environmental stressors through uspA gene expression were consistent with the behaviours observed in the response surface analyses. The uspA gene is a suitable biomarker for assessing environmental stressors of potential microorganisms for bioremediation of marine oil spills and for biosensing the ecophysiological status of native microbiota in a marine petroleum environment.

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Microbial remediation plays a pivotal role in the elimination of petroleum pollutants, making it imperative to investigate the capabilities of microorganisms in degrading petroleum. The present study describes the isolation of a promising strain, Acinetobacter sp. HX09, from petroleum-contaminated water. GC-MS analysis revealed a remarkable removal efficiency for short and medium-chain alkanes, with a rate of approximately 64% after a 7-days incubation at 30 °C. Transcriptome analysis of HX09 exhibited a predominant upregulation in gene expression levels by the induce of crude oil. Notably, genes such as alkane 1-monooxygenase, dehydrogenases and fatty acid metabolic enzymes exhibited fold changes range from 3.16 to 1.3. Based on the alkB gene sequences in HX09, the Phyre2 algorithm generated a three-dimensional structure that exhibited similarity to segments of acyl coenzyme desaturases and acyl lipid desaturases. Furthermore, three biodegradation-related gene clusters were predicted in HX09 based on the reference genome sequence. These findings contribute to our understanding of the hydrocarbon-degrading mechanisms employed by Acinetobacter species and facilitate the development of effective remediation strategies for crude oil- polluted environments.

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Oceanic oil spills present significant ecological risks that have the potential to contaminate extensive areas, including coastal regions. The occurrence of the 2019 oil spill event in Brazil resulted in over 3000 km of contaminated beaches and shorelines. While assessing the impact on benthic and beach ecosystems is relatively straightforward due to direct accessibility, evaluating the ecotoxicological effects of open ocean oil spills on the pelagic community is a complex task. Difficulties are associated with the logistical challenges of responding promptly and, in case of the Brazilian mysterious oil spill, to the subsurface propagation of the oil that impeded remote visual detection. An oceanographic expedition was conducted in order to detect and evaluate the impact of this oil spill event along the north-eastern Brazilian continental shelf. The pursuit of dissolved and dispersed oil compounds was accomplished by standard oceanographic methods including seawater polycyclic aromatic hydrocarbons (PAHs) analysis, biomass stable carbon isotope (δ13C), particulate organic carbon to particulate organic nitrogen (POC:PON) ratios, nutrient analysis and ecotoxicological bioassays using the naupliar phase of the copepod Tisbe biminiensis. Significant ecotoxicological effects, reducing naupliar development by 20-40 %, were indicated to be caused by the presence of dispersed oil in the open ocean. The heterogeneous distribution of oil droplets aggravated the direct detection and biochemical indicators for oil are presented and discussed. Our findings serve as a case study for identifying and tracing subsurface propagation of oil, demonstrating the feasibility of utilizing standard oceanographic and ecotoxicological methods to assess the impacts of oil spill events in the open ocean. Ultimately, it encourages the establishment of appropriate measures and responses regarding the liability and regulation of entities to be held accountable for oil spills in the marine environment.

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In this comprehensive study, we delved into the capabilities of five fungal strains: Aspergillus flavus, Aspergillus niger, Penicillium chrysogenum, Penicillium glabrum, and Penicillium rubens (the latter isolated from heavy crude oil [HCO]) in metabolizing HCO as a carbon source. Employing a meticulously designed experimental approach, conducted at room temperature (25 °C), we systematically explored various culture media and incubation periods. The results unveiled the exceptional resilience of all these fungi to HCO, with A. flavus standing out as the top performer. Notably, A. flavus exhibited robust growth, achieving a remarkable 59.1% expansion across the medium's surface, accompanied by distinctive macroscopic traits, including a cottony appearance and vibrant coloration. In an effort to further scrutinize its biotransformation prowess, we conducted experiments in a liquid medium, quantifying CO2 production through gas chromatography, which reached its zenith at day 30, signifying substantial bioconversion with a 38% increase in CO2 production. Additionally, we monitored changes in surface tension using the Du Noüy ring method, revealing a reduction in aqueous phase tension from 72.3 to 47 mN/m. This compelling evidence confirms that A. flavus adeptly metabolizes HCO to fuel its growth, while concurrently generating valuable biosurfactants. These findings underscore the immense biotechnological potential of A. flavus in addressing challenges related to HCO, thereby offering promising prospects for bioremediation and crude oil bioupgrading endeavors.

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Asphaltenes are the most polar and heavy fraction of petroleum, and their complex structure and toxicity make them resistant to biodegradation. The ability to tolerate high asphaltene concentrations is crucial to reducing the toxicity-related inhibition of microbial growth and improving their capacity for adaptation, survival, and biodegradation in soils highly contaminated with asphaltenes. This study developed a highly tolerant consortium for efficient asphaltene biodegradation in soils from 22 bacterial isolates obtained from heavy-crude oil-contaminated soils. Isolates corresponded to the Rhodococcus, Bacillus, Stutzerimonas, Cellulosimicrobium, Pseudomonas, and Paenibacillus genera, among others, and used pure asphaltenes and heavy crude oil as the only carbon sources. Surface plate assays were used to evaluate the tolerance of individual isolates to asphaltenes, and the results showed variations in the extension and inhibition rates with maximum tolerance levels at 60,000 mg asphaltenes l-1. Inhibition assays were used to select non-antagonistic bacterial isolates among those showing the highest tolerance levels to asphaltenes. A consortium made up of the five most tolerant and non-antagonistic bacterial isolates was able to degrade up to 83 wt.% out of 10,000 mg asphaltenes kg-1 in the soil after 52 days. Due to its biological compatibility, high asphaltene tolerance, and ability to utilise it as a source of energy, the degrading consortium developed in this work has shown a high potential for soil bioremediation and is a promising candidate for the treatment of aged soil areas contaminated with heavy and extra-heavy crude oil. This would be the first research to assess and consider extreme bacterial tolerance and microbial antagonism between individual degrading microbes, leading to the development of an improved consortium capable of efficiently degrading high amounts of asphaltenes in soil.

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The frequent occurrence of oil spills has led to serious environmental pollution and ecological issues. Given the high-viscosity of crude oil, it is essential to develop sorbents with efficient viscosity reduction and sorption capacity in various environmental conditions. Herein, a superhydrophobic carboxymethyl cellulose (CMC) aerogel co-modified by MXene and graphene jointly (M-Mxene/Gr CA) with aligned channels structure was prepared. The aligned channels structure can effectively improve the longitudinal thermal conductivity and reduce the sorption resistance. Through the modification of MXene and graphene, the aerogel realized efficient photo/electro-thermal conversion, thus ensuring its adaption to various working environments. The rapid heat generation can significantly reduce the viscosity of crude oil, achieving rapid recovery. Under one sun illumination (1.0 kW/m2), the surface temperature of M-Mxene/Gr CA can reach 72.6 °C and its sorption capability for high-viscous crude oil reaches 18 g/g. Combining photo-thermal and electro-thermal (0.5 kW/m2 and 23 V), the average sorption rate of crude oil can reach 1.3 × 107 g m-3 s-1. Finally, we present a continuous sorption system to recover offshore oil spills under the assistance of a pump. This work provides a new option for tackling high-viscous offshore oil spills due to its environmental friendliness and fast sorption capacity.

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Bacterial degradation of crude oil is a promising strategy for reducing the concentration of hydrocarbons in contaminated environments. In the first part of this study, we report the enrichment of two bacterial consortia from deep sediments of the Gulf of Mexico with crude oil as the sole carbon and energy source. We conducted a comparative analysis of the bacterial community in the original sediment, assessing its diversity, and compared it to the enrichment observed after exposure to crude oil in defined cultures. The consortium exhibiting the highest hydrocarbon degradation was predominantly enriched with Rhodococcus (75%). Bacterial community analysis revealed the presence of other hydrocarbonoclastic members in both consortia. In the second part, we report the isolation of the strain Rhodococcus sp. GOMB7 with crude oil as a unique carbon source under microaerobic conditions and its characterization. This strain demonstrated the ability to degrade long-chain alkanes, including eicosane, tetracosane, and octacosane. We named this new strain Rhodococcus qingshengii GOMB7. Genome analysis revealed the presence of several genes related to aromatic compound degradation, such as benA, benB, benC, catA, catB, and catC; and five alkB genes related to alkane degradation. Although members of the genus Rhodococcus are well known for their great metabolic versatility, including the aerobic degradation of recalcitrant organic compounds such as petroleum hydrocarbons, this is the first report of a novel strain of Rhodococcus capable of degrading long-chain alkanes under microaerobic conditions. The potential of R. qingshengii GOMB7 for applications in bioreactors or controlled systems with low oxygen levels offers an energy-efficient approach for treating crude oil-contaminated water and sediments.

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The current investigation was carried out to assess the potential of fungi isolated from polluted soil samples in Al Jubail, Saudi Arabia, to degrade crude oil. In a minimal salt medium with 1% crude oil as the carbon source, the growth potential of various fungal isolates was examined. Among twelve fungal isolates, YS-6 and YS-10, identified as Cunninghamella echinulata and Mucor circinelloides based on multiple sequence comparisons and phylogenetic analyses, were selected as having superior crude oil degrading abilities. To the best of our knowledge, the isolated species have never been detected in polluted soil samples in the eastern province of Saudi Arabia. YS-6 and YS-10 have shown their capacity to metabolize crude oil by removing 59.7 and 78.1% of crude oil, respectively. Interestingly, they succeeded in reducing the surface tension to 41.2 and 35.9 mN/m, respectively. Moreover, the emulsification activity and hydrophobicity were determined to be 36.7, 44.9, 35.9, and 53.4%, respectively. The recovery assays included zinc sulfate, ammonium sulfate, acid precipitation, and solvent extraction techniques. All these approaches showed that the amount of biosurfactants correlates to the tested hydrocarbons. Furthermore, the enzyme activity of these two isolates generated significantly more laccase (Lac) than manganese peroxidase (MnP) and lignin peroxidase (LiP), as compared to the control. In conclusion, our study highlights new perspectives on the fungal resources found in persistently polluted terrestrial ecosystems. This knowledge will be useful for bioremediation, safe disposal of petroleum-oil contamination, and other industrial uses.

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Small wild mammals have been used to measure the damage caused by exposure to oil-contaminated soil, including deer mice. However, the study of toxic effects of crude oil using oxidative damage biomarkers in the wild rodent Calomys laucha (Vesper mouse) is absent. This investigation aimed to evaluate the effects of acute exposure to contaminated soil with different concentrations of crude oil (0, 1, 2, 4 and 8% w/w), simulating an accidental spill, using oxidative stress biomarkers in the liver, kidneys, lungs, testes, paw muscle, and lymphocytes of C. laucha. Animals exposed to the contaminated soil showed increases in lipid peroxidation and protein carbonylation at the highest exposure concentrations in most organ homogenates analyzed and also in blood cells, but responses to total antioxidant capacity were tissue-dependent. These results showed that acute exposure to oil-contaminated soil caused oxidative damage in C. laucha and indicate these small mammals may be susceptible to suffer the impacts of such contamination in its occurrence region, threatening the species' survival.

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The objective of this study is to examine the saturation process in a column containing Brazil nuts and possible changes in the quality of the product. Brazil nut samples were initially placed in a cylindrical PVC column 15 cm in diameter and 110 cm in height. The ozone gas concentrations of 2.5, 4.5, 9.0, and 14.0 mg L-1 and a flow rate of 3.0 L min-1 were applied at a temperature of 25 ºC. Ozone gas was injected at the base of the cylindrical column, and the seed column height values adopted were 0.25, 0.50, and 0.75 m. Saturation concentration and time were determined. To measure possible changes in the quality of ozonized Brazil nuts, moisture and color, as well as qualitative variables of the crude oil were evaluated at the exposure times of 0, 3, 6, 9, and 12 h. To evaluate the quality of the crude oil extracted from ozonized nuts, the free fatty acid content, peroxide value, and iodine value were analyzed. Increasing ozonation times increased ozone concentration at all inlet gas concentrations. Saturation time decreased as the inlet gas concentration was increased, at the different product column heights. There was no change in product moisture in response to ozonation. Ozonation did not induce significant changes in color or in the crude oil, due to the triple interaction between column height, ozone concentration, and exposure time. In conclusion, the height of the product's column influences saturation time and concentration during the ozonation process. Considering the color of the product and characteristics of its crude oil, the use of ozone under the conditions adopted in the present study does not affect the quality of Brazil nuts to the point of rendering them unmarketable.

O objetivo do presente trabalho é estudar o processo de saturação em coluna contendo castanha-do-Brasil e possíveis alterações na qualidade do produto. Inicialmente as amostras de castanha-do-Brasil foram acondicionadas em coluna cilíndrica de PVC de 15 cm de diâmetro e 110 cm de altura. Foram adotadas as concentrações do gás ozônio de 2,5, 4,5, 9,0 e 14,0 mg L-1 e vazão de 3,0 L min-1, na temperatura de 25 ºC. O gás ozônio foi injetado na base da coluna cilíndrica e os valores adotados de altura da coluna de grãos foram de 0,25, 0,50, e 0,75 m. Determinaram-se o tempo e a concentração de saturação. Na avaliação de possíveis alterações na qualidade de castanhas-do-Brasil ozonizadas foram determinados a umidade, coloração e variáveis qualitativas do óleo bruto, com tempos de exposição de 0, 3, 6, 9 e 12 h. Para avaliação da qualidade do óleo bruto extraído de castanhas ozonizadas foram analisadas o teor de ácidos graxos livres, o índice de peróxido e o índice de iodo. A elevação do período de ozonização promoveu aumento da concentração do ozônio para todas as concentrações de entrada do gás. No que se refere aos valores de tempo de saturação, à medida que se elevou a concentração de entrada do gás, houve redução do tempo de saturação, nas diferentes alturas de coluna do produto. Não houve variação da umidade do produto em decorrência da ozonização. A ozonização não provocou alterações significativas na cor e no óleo bruto, em decorrência da interação tripla entre altura da coluna do produto, concentração do ozônio e tempo de exposição. É possível concluir que a altura da coluna do produto influencia o tempo e a concentração de saturação, durante o processo de ozonização. O uso do ozônio nas condições adotadas no presente estudo não afeta a qualidade da castanhado-Brasil, considerando-se a cor do produto e características do óleo bruto, de tal forma a inviabilizar a comercialização.

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Research reports reveal the importance of applying various substances to enhance extra-heavy crude oil pipeline transportation. During the crude oil conduction process, shearing occurs in the equipment and pipe accessories, producing a water-in-crude emulsion associated with forming a rigid film by adsorbing natural surfactant molecules in the droplets water, leading to increased Viscosity. This study presents the effect of a flow enhancer (FE) on the behavior of the Viscosity of an extra heavy crude oil (EHCO) and in emulsions formed with 5% and 10% water (W). The results revealed the effectiveness of the 1%, 3%, and 5% flow enhancer in lowering the Viscosity and presenting a Newtonian flow behavior which will help reduce the cost of heat treatment during the transportation of crude oil through the pipeline.

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In this study, a miniaturized liquid-liquid extraction (LLE) method for pre-concentration of Na, K, Ca, and Mg in crude oil was proposed. Analytes in crude oil were quantitatively extracted to the aqueous phase, followed by flame atomic absorption spectrometry (FAAS) determination. The following parameters were evaluated: type of extraction solution, sample mass, heating temperature and time, stirring time, centrifugation time, and the use of toluene and chemical demulsifier. Accuracy was evaluated by comparing the results obtained by the proposed method (LLE-FAAS) with those obtained after high-pressure microwave-assisted wet digestion and FAAS determination (reference values). No statistical difference was observed between the reference values and those using the optimized conditions for LLE-FAAS: 2.5 g of sample; 1000 µL of 2 mol L-1 HNO3, 50 mg L-1 of chemical demulsifier in 500 µL of toluene, 10 min of heating at 80 °C, 60 s of stirring, and 10 min of centrifugation. Relative standard deviations were lower than 6%. The limits of quantification (LOQ) were 1.2, 1.5, 5.0, and 0.50 µg g-1 for Na, K, Ca, and Mg, respectively. The proposed miniaturized LLE method presents several advantages, such as ease-of-use, high throughput (up 10 samples can be processed per 1 h), uses a high sample mass reaching low LOQs. In addition, the use of a diluted solution for extraction reduces the amount of reagents (around 40 times) and consequently laboratory residue generation, becoming an environmental friendly method. Suitable LOQs were achieved for analyte determination at low concentration even using a simple and low-cost sample preparation system (miniaturized LLE method) and a relatively low-cost determination technique (FAAS), avoiding the use of microwave ovens and more sensitivity techniques, which are required for routine analyses.

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Crude oil distillates are a highly useful industrial product, mainly for energy generation. Unfortunately, they are rarely studied, mainly due to the low accessibility to products directly obtained from the distillation process, which is a laborious, expensive, and time-consuming operation. This work presents and discusses the use of time-domain nuclear magnetic resonance (TD-NMR) as a simple, affordable, and straightforward tool for the development of correlations supported on the transverse relaxation time (T2 ) and boiling temperature. The results point out a high convergence between TD-NMR experimental data and the ASTM D2892 method for distillates from light, medium, and heavy oils, with up to 52.20% of accumulated mass and boiling point temperature (Tb ) up to 400°C. Furthermore, an unprecedented relationship between T2 values and the accumulated mass of the distillates is first demonstrated. This new insight opens new perspectives for future prediction of accumulated mass for unknown crude oils, placing the TD-NMR relaxometry as an appeal spectroscopy approach with a potential to meaningfully contribute to the daily refining petrochemical industry field operations.

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This study aims to develop and evaluate fracturing nanofluids from the laboratory to the field trial with the dual purpose of increasing heavy crude oil mobility and reducing formation damage caused by the remaining fracturing fluid (FF). Two fumed silica nanoparticles of different sizes, and alumina nanoparticles were modified on the surface through basic and acidic treatments. The nanoparticles were characterized by transmission electron microscopy, dynamic light scattering, zeta potential and total acidity. The rheological behavior of the linear gel and the heavy crude oil after adding different chemical nature nanoparticles were measured at two concentrations of 100 and 1000 mg/L. Also, the contact angle assessed the alteration of the rock wettability. The nanoparticle with better performance was the raw fumed silica of 7 nm at 1000 mg/L. These were employed to prepare a fracturing nanofluid from a commercial FF. Both fluids were evaluated through their rheological behavior as a function of time at high pressure following the API RP39 test, and spontaneous imbibition tests were carried out to assess the FF's capacity to modify the wettability of the porous media. It was possible to conclude that the inclusion of 7 nm commercial silica nanoparticles allowed obtaining a reduction of 10 and 20% in the two breakers used in the commercial fracture fluid formulation without altering the rheological properties of the system. Displacement tests were also performed on proppant and rock samples at reservoir conditions of overburden and pore pressures of 3200 and 1200 psi, respectively, while the temperature was set at 77 °C and the flow rate at 0.3 cm3/min. According to the effective oil permeability, a decrease of 31% in the damage was obtained. Based on these results, the fracturing nanofluid was selected and used in the first worldwide field application in a Colombian oil field with a basic sediment and water (BSW%) of 100 and without oil production. After two weeks of the hydraulic fracture operation, crude oil was produced. Finally, one year after this work, crude oil viscosity and BSW% kept showing reductions near 75% and 33%, respectively; and having passed two years, the cumulative incremental oil production is around 120,000 barrels.

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During the oil production and processing stages, the asphaltene precipitation is one of the great operation problems of oil industry. It can precipitate in the formation, tubing, or surface, causing operating problems, such as reduction in oil recovery by changing the reservoir permeability and wettability, clogging of the pipelines, and difficulty in separations process. The quantification of asphaltenes in petroleum by ASTM D6560 standard method is very laborious and use of a larger solvent volume than necessary. The present work proposes the use of time domain nuclear magnetic resonance (TD-NMR) as new methodology to quantify the asphaltene precipitated in crude oil. Three (light, medium, and heavy) crude oils with asphaltenes content of 0.97, 1.88, and 7.00 wt% were mixed with n-heptane in different R (ml of solvent/g of oil) values and analyzed by means of transverse relaxation time (T2 ). According NMR results, the R values enough for complete asphaltene precipitation for the oils A, B, and C were, respectively, equal to 16.50, 23.00, and 39.50 ml g-1 . These outcomes represent a reduction of 58.75%, 42.50%, and 1.25% in the solvent volume per mass of oil for the oil A, B, and C, respectively, compared to the ASTM D6560 method, which imposes 40 ml g-1 . Therefore, it has been shown that TD-NMR can be applied to estimate the amount of asphaltene precipitated in petroleum and have potential to be applied in routine analysis with advantages of saving time and costs.

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This work investigates the impact of the Covid-19 outbreak on crude oil market efficiency. The approach is based on the singular value decomposition (SVD) entropy. Iso-distributional surrogate data test was used to contrast the results against random patterns, and phase randomization based on Fourier transform was used to assess nonlinearities. The analysis considered the WTI market and focused on the Covid-19 pandemic period January 2020-November 2021 and contrasted with the long preceding period from January 2000 to date. It was found that the crude oil market was informationally efficient most of the time with small sporadic deviations from efficiency in the pre-Covid-19 years. The Covid-19 period exhibited the largest deviations from efficiency, mainly in the first months of the outbreak, accompanied by a marked reduction of nonlinear components. The analysis was conducted for different scales, and the results showed that the deviations from efficiency were more pronounced for quarterly scales. For the sake of comparison, the analysis was also carried out on the trading volume dynamics and the results showed that the market activity is not fully random. The dynamics of the trading volume exhibited significant deviations from the randomness behavior when the crude oil market was efficient, and a behavior that was consistent with nonlinear patterns. The opposite behavior was noted for stages when the crude oil market showed strong deviations from efficiency. Overall, the findings of this study suggest an increasing opportunity for crude oil price predictions and abnormal returns during the Covid-19 pandemic.

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This study provides the first assessment of the impact of the worst oil spill ever faced by Brazilian sandy beaches by analyzing changes in the community structure of intertidal macroinfauna. Four sandy beaches were sampled three times after the oil spill, and the results were compared with previous data. The first sampling, conducted 15 days after the oil spill (October 2019), showed higher abundance of macroinfauna, which decreased in subsequent sampling conducted 60 and 120 days later, but never reached a lower level than in previous sampling. Of the macroinfauna species, Scolelepis sp. was most abundant in October 2019, while Donax gemmula was predominant during the other time periods. Changes observed in macroinfauna were due to natural fluctuations rather than a response to this disturbance. Characteristics of both the oil spill and macroinfauna contributed to the lack of acute impacts.

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The effect of abiotic factors, such as weather and climate can alter the properties of vegetable oils. In this work, the effects of the refining process and pre-drying of Moringa oleifera Lam. seeds collected in the dry and rainy seasons (seasonality) were evaluated on the characteristics of the extracted oils. The refined and crude dry season oils had lower acidity and moisture content than the dry and raw rainy season oils. Oleic acid (C18:1) showed the highest concentration in the different Moringa oils studied. The results suggest that dry season oils are more suitable for application as feedstocks in the cosmetic, food, medicinal and pharmaceutical industries. Furthermore, refining proved to be efficient in removing free fatty acids and in lightening the oil.

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Fish development can be affected by environmental pollutants such as crude oil (anthropogenic or natural sources), causing alterations especially in cardiac function and morphology. Most such studies have focused on saltwater species, whereas studies in freshwater fishes are scant. The objective of the current study was to evaluate the effects of crude oil exposure (as 0, 5, 10, 15, or 20% high-energy water accommodated fractions, HEWAF) on cardiac function and edema formation during two early periods of development (embryo and eleuteroembryo, 48 h each) individually using the tropical gar Atractosteus tropicus as a model. Embryos did not exhibit alterations in body mass, total length, condition factor, and cardiac function as a function of oil. In contrast, eleuteroembryos proved to be more sensitive and exhibited increased body mass, total length, and condition factor, decreased heart rate and phenotypic alterations such as cardiac dysmorphia (tubular hearts) and spine curvature at high concentrations of HEWAF. Moreover, edema formation was observed in both stages This study shows different functional responses of A. tropicus after crude oil exposure and provides useful information of the developmental impacts of these compounds on the early life stages of freshwater tropical fishes.

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In 2018 we evaluated at 48 h and 96 h, the gene expression profile of larvae of Limulus polyphemus exposed to 10% and 100% of water-accommodated fraction (WAF) of light crude oil (API 35), and 10% and 100% of a chemically enhanced WAF (CEWAF) with the dispersant Nokomis 3-F4® in a static-acute (96 h) bioassay. Alkanes and PAHs concentrations were higher in CEWAF than in WAF stock solutions. Under the proved conditions, the expression profile of genes associated to detoxification processes (glutathione S-transferase and glutathione peroxidase), stress (heat shock protein), innate immunity (tumor necrosis factor receptor-associated factor 4 traf4), cell death (apoptosis inhibitor 5) and DNA repairing (E3 ubiquitin protein ligase), showed a deregulation at 48 h followed by an upregulation at 96 h, with exception of glutathione peroxidase, heat shock protein and innate immunity that remained low in CEWAF. In conclusion, by using genes that have been proposed as biomarkers to pollutants exposure, L. polyphemus larvae showed an early activation of genes related to the immune system, antioxidant, heat shock and NER.

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