Prenatal exposure of bonny light crude oil induces embryotoxicity, impaired cognitive functions and cortico-hippocampal neurodegeneration on fetal outcomes of pregnant sprague-dawley rats.

The low Sulfur level, heavy metals and easy production rate of Bonny Light Crude Oil (BLCO) makes it one of Nigeria's most explored oil. This study investigated the memory impairments, embryotoxicity and cortico-hippocampal neurodegeneration induced by prenatal exposure to BLCO of pregnant Sprague-Dawley (S-D) rats. Twenty pregnant rats were divided into 4 groups (A-D) of 5 rats each. Group A received normal saline as placebo. Group B-D received oral doses of BLCO at 0.73 ml/kg, 2 ml/kg and 3.8 ml/kg on pregnancy day 8-12.5 respectively. The pregnant rats were allowed to litter and nurse their pups. At 6 weeks postnatal life, twelve (12) selected young rats (n = 12) were accessed for behavioral study (Y-maze) and then sacrificed for biochemical and histological analysis. The results showed spontaneous abortion, still births and significantly reduced number of live births in the high dose group of BLCO compared to control. Length of gestation was significantly increased in the high dose group when compared to the control. CAT levels reduced significantly with concomitant increase in 8-OHdG among BLCO treated groups compared to control. Spontaneous alteration and number of arm entries decreased in the BLCO groups in comparison to control. Histological observation showed reduced cellular size, chromatolysis and presence of extracellular senile plaques in the prefrontal cortex and mild histological changes in the hippocampus architecture in the BLCO treated groups compared to the control. BLCO is capable of inducing embryotoxicity, impair cognition and cortico-hippocampal neurodegeneration.

Optimization of the Biodegradation of Aliphatic, Aromatic, and UCM Hydrocarbons from Light Crude Oil in Marine Sediment Using Response Surface Methodology (RSM).

This study describes the optimization of the biodegradation of total aliphatic (tAHCs), total aromatic (tPAHs), and unresolved complex mixture (UCM) hydrocarbons from light crude oil in marine sediment. The response surface methodology (RSM), with a Box-Behnken design, was used to optimize the hydrocarbon fraction degradation, reported as degradation efficiency (E (%)), using four independent variables (inoculum, dispersant, light oil concentration, and carbon/nitrogen ratio), all at three levels. Analysis of variance (ANOVA) showed R2 values of 0.976, 0.974, and 0.975 for tAHCs, tPAHs, and UCM, respectively. All fractions exhibited a statistically significant effect (P < 0.05) in the second-order quadratic regression model for degradation. According to the models, the optimal degradation prediction was: 81.03% for tAHCs, 85.96% for tPAHs, and 92.86% for UCM. This work highlights the possibility of carrying out efficient biodegradation, of more than 80%, through an optimization process using different light oil concentrations, opening up possibilities of multiple response optimization.

Effects of a Light Crude Oil Spill on a Tropical Coastal Phytoplankton Community.

Oiling scenarios following spills vary in concentration and usually can affect large coastal areas. Consequently, this research evaluated different crude oil concentrations (10, 40, and 80 mg L-1) on the nearshore phytoplanktonic community in the southern Gulf of Mexico. This experiment was carried out for ten days using eight units of 2500 L each; factors monitored included shifts in phytoplankton composition, physicochemical parameters and the culturable bacterial abundance of heterotrophic and hydrocarbonoclastic groups. The temperature, salinity, and nutrient concentrations measured were within the ranges previously reported for Yucatan Peninsula waters. The total hydrocarbon concentration (TPH) in the control at T0 indicated the presence of hydrocarbons (PAHs 0.80 µg L-1, aliphatics 7.83 µg L-1 and UCM 184.09 µg L-1). At T0, the phytoplankton community showed a similar assemblage structure and composition in all treatments. At T10, the community composition remained heterogeneous in the control, in agreement with previous reports for the area. However, for oiled treatments, Bacillariophyceae dominated at T10. Hydrocarbonoclastic bacteria were associated with oiled treatments throughout the experiment, while heterotrophic bacteria were associated with control conditions. Our results agreed with previous works at the taxonomic level showing the presence of Bacillariophyceae and Dinophyceae in oil-related treatments, where these groups showed the major interactions in co-occurrence networks. In contrast, Chlorophyceae showed the key node in the co-occurrence network for the control. This study aims to contribute to knowledge on phytoplankton community shifts during a crude oil spill in subtropical oligotrophic regions.

Differential Gene Expression Induced by Acute Exposure to Water Accommodated Fraction (WAF) and Chemically Enhanced WAF (CEWAF) of Light Crude Oil and Nokomis 3-F4 in Limulus polyphemus Larvae.

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.

Bacterial succession and co-occurrence patterns of an enriched marine microbial community during light crude oil degradation in a batch reactor.

AIMS: The aim of this study was to investigate the dynamic changes in the bacterial structure and potential interactions of an acclimatized marine microbial community during a light crude oil degradation experiment. METHODS AND RESULTS: The bacterial community effectively removed 76·49% of total petroleum hydrocarbons after 30 days, as evidenced by GC-FID and GC-MS analyses. Short-chain alkanes and specific aromatic compounds were completely degraded within the first 6 days. High-throughput sequencing of 16S rRNA gene indicated that the starting bacterial community was mainly composed by Marinobacter and more than 30 non-dominant genera. Bacterial succession was dependent on the hydrocarbon uptake with Alcanivorax becoming dominant during the highest degradation period. Sparse correlations for compositional data algorithm revealed one operational taxonomic unit (OTU) of Muricauda and an assembly of six OTUs of Alcanivorax dieselolei and Alcanivorax hongdengensis as critical keystone components for the consortium network maintenance and stability. CONCLUSIONS: This work exhibits a stabilized marine bacterial consortium with the capability to efficiently degrade light crude oil in 6 days, under laboratory conditions. Successional and interaction patterns were observed in response to hydrocarbon consumption, highlighting potential interactions between Alcanivorax and keystone non-dominant OTUs over time. SIGNIFICANCE AND IMPACT OF THE STUDY: Our results contribute to the understanding of interactions and potential roles of specific members of hydrocarbonoclastic marine bacterial communities, which will be useful for further bioaugmentation studies concerning the associations between indigenous and introduced micro-organisms.

Quercetin and vitamin E attenuate Bonny Light crude oil-induced alterations in testicular apoptosis, stress proteins and steroidogenic acute regulatory protein in Wistar rats.

Studies have shown the reproductive effects of Bonny Light crude oil (BLCO) via the mechanism of oxidative stress and testicular apoptosis. We investigated the protective role of quercetin and vitamin E on BLCO-induced testicular apoptosis. Experimental rats were divided into four groups of four each. Animals were orally administered 2 ml/kg corn oil (control: group 1), BLCO-800 mg/kg body weight + 10 mg/kg quercetin (group 2), BLCO-800 mg/kg body weight + 50 mg/kg vitamin E (group 3) and BLCO-800 mg/kg body weight only (group 4) for 7 d. Protein levels of caspase 3, FasL, NF-kB, steroidogenic acute regulatory protein and stress response proteins were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Immunofluorescence staining was used to quantify the expression of caspase 3, FasL and NF-kB. Apoptosis was quantified by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) assay. Administration of BLCO resulted in a significant increase in the levels of stress response proteins and apoptosis-related proteins by 50% and above after 7 d following BLCO exposure and a concomitant increase in expression of caspase 3, FasL and NF-kB expression by immunofluorescence staining. Apoptosis showed a significant increase in TUNEL positive cells. Co-administration with quercetin or vitamin E reversed BLCO-induced apoptosis and levels of stress protein, relative to control. These findings suggest that quercetin and vitamin E may confer protection against BLCO-induced testicular oxidative stress-related apoptosis.

Transient effect of single dose exposure of Nigerian Bonny-light crude oil on testicular steroidogenesis in Wistar rats is accompanied by oxidative stress.

The folkloric use of Nigerian Bonny-light crude oil (BLCO) in Niger Delta area of Nigeria is a common practice. There is increasing experimental evidence portending the adverse effects of BLCO an environmental toxicant on testicular function. We investigated the effects of single dose of BLCO (800 mg/kg body weight) on the activities of steroidogenic and antioxidant enzymes such as serum follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone, 3 ß-hydroxy-steroid dehydrogenase (3 ß-HSD), 17 ß-hydroxy-steroid dehydrogenase (17 ß-HSD), superoxide dismutase (SOD), glutathione-S-transferase (GST) and glutathione peroxidase (GSH-Px), levels of lipid peroxidation (LPO), glutathione reduced (GSH) and steroidogenic acute regulatory (StAR) protein, in testes of rats. There was a sequential reduction in the concentration of steroid hormones and activities of steroidogenic enzymes with a concomitant decrease in levels of StAR protein, followed by a parallel increase in antioxidant enzyme activities and levels of LPO. These findings revealed inhibitory effects of BLCO on testicular steroidogenesis and the possible role of oxidative stress in testicular dysfunction observed in this study.

Vicia faba seed: a bioindicator of phytotoxicity, genotoxicity, and cytotoxicity of light crude oil.

Crude oil contamination is a serious threat to the environment and human health as it can contaminate food chains. Therefore, it is necessary to find efficient tests to monitor soils for crude oil contamination. The present study investigates the efficacy of Vicia faba seeds for monitoring contaminated soils with light crude oil. Vicia faba seeds were planted in 0 (control), 1, 2, and 4 percentages (weight percentage) light crude oil-contaminated soils. The seed germination and root length were measured to evaluate phytotoxicity, while the mitotic index, chromosome aberrations, and micronucleus formation in the root tip cells were examined for cytotoxicity and genotoxicity tests. The results showed that light crude oil had toxic effects on Vicia faba growth characteristics, even at 1% contamination. The phytotoxicity assay showed that crude oil reduced seed germination and root length by 45% and 61.67%, respectively. In contrast, cellular observations indicated an increase in mitotic index, chromosome aberrations, and micronucleus formation up to 3, 3.59, and 5.6 times, respectively, compared to the control. The light crude oil at 4% induced the simultaneous occurrence of nuclear bud, polyploidy, and micronucleus that may be considered as severe clastogenic and aneugenic effects. Accordingly, Vicia faba can be considered a reliable living system for monitoring light crude oil pollution in soils, even at low concentrations.

Gill and liver transcriptomic responses of Achirus lineatus (Neopterygii: Achiridae) exposed to water-accommodated fraction (WAF) of light crude oil reveal an onset of hypoxia-like condition.

Crude oil is one of the most widespread pollutants released into the marine environment, and native species have provided useful information about the effect of crude oil pollution in marine ecosystems. We consider that the lined sole Achirus lineatus can be a useful monitor of the effect of crude oil in the Gulf of Mexico (GoM) because this flounder species has a wide distribution along the GoM, and its response to oil components is relevant. The objective of this study was to compare the transcriptomic changes in liver and gill of adults lined sole fish (Achirus lineatus) exposed to a sublethal acute concentration of water-accommodated fraction (WAF) of light crude oil for 48 h. RNA-Seq was performed to assess the transcriptional changes in both organs. A total of 1073 differentially expressed genes (DEGs) were detected in gills; 662 (61.69%) were upregulated, and 411 (38.30%) were downregulated whereas in liver, 515 DEGs; 306 (59.42%) were upregulated, and 209 (40.58%) were downregulated. Xenobiotic metabolism and redox metabolism, along with DNA repair mechanisms, were activated. The induction of hypoxia-regulated genes and the generalized regulation of multiple signaling pathways support the hypothesis that WAF exposition causes a hypoxia-like condition.

Changes in the Bacterioplankton Community Structure from Southern Gulf of Mexico During a Simulated Crude Oil Spill at Mesocosm Scale.

The southern Gulf of Mexico (sGoM) is highly susceptible to receiving environmental impacts due to the recent increase in oil-related activities. In this study, we assessed the changes in the bacterioplankton community structure caused by a simulated oil spill at mesocosms scale. The 16S rRNA gene sequencing analysis indicated that the initial bacterial community was mainly represented by Gamma-proteobacteria, Alpha-proteobacteria, Flavobacteriia, and Cyanobacteria. The hydrocarbon degradation activity, measured as the number of culturable hydrocarbonoclastic bacteria (CHB) and by the copy number of the alkB gene, was relatively low at the beginning of the experiment. However, after four days, the hydrocarbonoclastic activity reached its maximum values and was accompanied by increases in the relative abundance of the well-known hydrocarbonoclastic Alteromonas. At the end of the experiment, the diversity was restored to similar values as those observed in the initial time, although the community structure and composition were clearly different, where Marivita, Pseudohongiella, and Oleibacter were detected to have differential abundances on days eight-14. These changes were related with total nitrogen (p value = 0.030 and r2 = 0.22) and polycyclic aromatic hydrocarbons (p value = 0.048 and r2 = 0.25), according to PERMANOVA. The results of this study contribute to the understanding of the potential response of the bacterioplankton from sGoM to crude oil spills.

Bioremediation of oil-contaminated water by bacterial consortium immobilized on environment-friendly biocarriers

BACKGROUND: Accidental oil spills in the Nile River represents a public health concern in Egypt. Bioremediation of oil-contaminated water is efficiently competitive and economically effective compared to mechanical and chemical methods. AIM: The current study aimed to: i) assess using environment-friendly materials as carriers for immobilizing oil-degrading bacteria and ii) evaluate applying these biocarriers for removal of oil from water in a batch experiment. MATERIALS AND METHODS: Four bacterial strains Pseudomonas aeruginosa, Cronobacter sakazakii (recently named Enterobacter sakazakii), Klebsiella oxytoca and Bordetella bronchispetica were immobilized on grinded corn qgualh, Egyptian loofah, palm leaf raffia and sponge. Immobilizing bacterial consortium was carried out by incubating bacteria with carriers at 37°C with shaking at 120 rpm for three days. Biofilm formed on biocarriers was checked using scanning electron microscopy. Biocarriers with immobilized bacterial consortium were used for the removal of oil. Experiments were carried out in flasks each containing 100 ml of mineral salt broth fortified with 1 g/l light crude oil. Flasks were incubated at 37°C with shaking at 120 rpm for nine weeks. Total viable bacterial count and oil and grease were determined weekly. RESULTS: Oil and grease measurements showed a significant removal that reached 68.3, 71.5, 77.7 and 81% for grinded corn qgualh, Egyptian loofah, palm leaf raffia and sponge respectively. Flasks inoculated with sponge biocarriers showed rapid oil removal compared with other biocarriers. Gas chromatography-mass spectrometry (GC-MS) showed a percentage removal that reached 87% for compounds C11 to C32 in case of sponge carrier. CONCLUSION: Immobilizing bacteria on biocarriers showed a good biofilm formation. Immobilized bacterial consortium showed rapid and efficient oil removal in bioremediation processes. Sponge showed dense bacterial colonization and consequently biofilm formation compared with other biocarriers.

Exposures of zebrafish through diet to three environmentally relevant mixtures of PAHs produce behavioral disruptions in unexposed F1 and F2 descendant.

The release of polycyclic aromatic hydrocarbons (PAHs) into the environment has increased very substantially over the last decades. PAHs are hydrophobic molecules which can accumulate in high concentrations in sediments acting then as major secondary sources. Fish contamination can occur through contact or residence nearby sediments or though dietary exposure. In this study, we analyzed certain physiological traits in unexposed fish (F1) issued from parents (F0) exposed through diet to three PAH mixtures at similar and environmentally relevant concentrations but differing in their compositions. For each mixture, no morphological differences were observed between concentrations. An increase in locomotor activity was observed in larvae issued from fish exposed to the highest concentration of a pyrolytic (PY) mixture. On the contrary, a decrease in locomotor activity was observed in larvae issued from heavy oil mixture (HO). In the case of the third mixture, light oil (LO), a reduction of the diurnal activity was observed during the setup of larval activity. Behavioral disruptions persisted in F1-PY juveniles and in their offspring (F2). Endocrine disruption was analyzed using cyp19a1b:GFP transgenic line and revealed disruptions in PY and LO offspring. Since no PAH metabolites were dosed in larvae, these findings suggest possible underlying mechanisms such as altered parental signaling molecule and/or hormone transferred in the gametes, eventually leading to early imprinting. Taken together, these results indicate that physiological disruptions are observed in offspring of fish exposed to PAH mixtures through diet.

BIOB: a mathematical model for the biodegradation of low solubility hydrocarbons.

Modeling oil biodegradation is an important step in predicting the long term fate of oil on beaches. Unfortunately, existing models do not account mechanistically for environmental factors, such as pore water nutrient concentration, affecting oil biodegradation, rather in an empirical way. We present herein a numerical model, BIOB, to simulate the biodegradation of insoluble attached hydrocarbon. The model was used to simulate an experimental oil spill on a sand beach. The biodegradation kinetic parameters were estimated by fitting the model to the experimental data of alkanes and aromatics. It was found that parameter values are comparable to their counterparts for the biodegradation of dissolved organic matter. The biodegradation of aromatics was highly affected by the decay of aromatic biomass, probably due to its low growth rate. Numerical simulations revealed that the biodegradation rate increases by 3-4 folds when the nutrient concentration is increased from 0.2 to 2.0 mg N/L.