Biochemical, molecular, and physiological assessments of crude oil dietary exposure in sub-adult red drum (Sciaenops ocellatus).

A 14-day exposure study in which sub-adult red drum (Sciaenops ocellatus) were fed a petroleum crude oil-treated pellet feed was conducted to assess the potential effects of ingesting an oil-contaminated food source. Though food consumption decreased, significant polycyclic aromatic hydrocarbons accumulated in the body and liver, which did not affect the body and liver's fatty acid composition. In the red drum given the crude oil-treated feed, a significant decrease in the RNA:DNA growth rate index was noted, while only subtle changes in body and liver lipid composition were seen. Differentially expressed gene analysis in the liver demonstrated a significant down-regulation of leptin and up-regulation of the aryl hydrocarbon receptor nuclear translocator-like protein 1. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses indicated enrichment of terms and pathways associated with cholesterol biosynthesis and oxidative stress. Ingenuity Pathway Analysis further predicted activation of seven pathways associated with cholesterol biosynthesis. Measured oxidative stress biomarkers in the blood indicated decreased systemic antioxidants with increased lipid peroxidation. The results of this study suggest that dietary oil exposure alters the signaling of biological pathways critical in cholesterol biosynthesis and disruptions in systemic oxidative homeostasis.

Dispersed Crude Oil Induces Dysbiosis in the Red Snapper Lutjanus campechanus External Microbiota.

The fish external microbiota competitively excludes primary pathogens and prevents the proliferation of opportunists. A shift from healthy microbiota composition, known as dysbiosis, may be triggered by environmental stressors and increases host susceptibility to disease. The Deepwater Horizon (DWH) oil spill was a significant stressor event in the Gulf of Mexico. Despite anecdotal reports of skin lesions on fishes following the oil spill, little information is available on the impact of dispersed oil on the fish external microbiota. In this study, juvenile red snapper (Lutjanus campechanus) were exposed to a chemically enhanced water-accommodated fraction (CEWAF) of Corexit 9500/DWH oil (CEWAF) and/or the bacterial pathogen Vibrio anguillarum in treatments designed to detect changes in and recovery of the external microbiota. In fish chronically exposed to CEWAF, immunoglobulin M (IgM) expression significantly decreased between 2 and 4 weeks of exposure, coinciding with elevated liver total polycyclic aromatic hydrocarbons (PAHs). Dysbiosis was detected on fish chronically exposed to CEWAF compared to seawater controls, and addition of a pathogen challenge altered the final microbiota composition. Dysbiosis was prevented by returning fish to clean seawater for 21 days after 1 week of CEWAF exposure. Four fish exhibited lesions during the trial, all of which were exposed to CEWAF but not all of which were exposed to V. anguillarum. This study indicates that month-long exposure to dispersed oil leads to dysbiosis in the external microbiota. As the microbiota is vital to host health, these effects should be considered when determining the total impacts of pollutants in aquatic ecosystems. IMPORTANCE Fish skin is an immunologically active tissue. It harbors a complex community of microorganisms vital to host homeostasis as, in healthy fish, they competitively exclude pathogens found in the surrounding aquatic environment. Crude oil exposure results in immunosuppression in marine animals, altering the relationship between the host and its microbial community. An alteration of the healthy microbiota, a condition known as dysbiosis, increases host susceptibility to pathogens. Despite reports of external lesions on fishes following the DWH oil spill and the importance of the external microbiota to fish health, there is little information on the effect of dispersed oil on the external microbiota of fishes. This research provides insight into the impact of a stressor event such as an oil spill on dysbiosis and enhances understanding of long-term sublethal effects of exposure to aid in regulatory decisions for protecting fish populations during recovery.

A review of the toxicology of oil in vertebrates: what we have learned following the Deepwater Horizon oil spill.

In the wake of the Deepwater Horizon (DWH) oil spill, a number of government agencies, academic institutions, consultants, and nonprofit organizations conducted lab- and field-based research to understand the toxic effects of the oil. Lab testing was performed with a variety of fish, birds, turtles, and vertebrate cell lines (as well as invertebrates); field biologists conducted observations on fish, birds, turtles, and marine mammals; and epidemiologists carried out observational studies in humans. Eight years after the spill, scientists and resource managers held a workshop to summarize the similarities and differences in the effects of DWH oil on vertebrate taxa and to identify remaining gaps in our understanding of oil toxicity in wildlife and humans, building upon the cross-taxonomic synthesis initiated during the Natural Resource Damage Assessment. Across the studies, consistency was found in the types of toxic response observed in the different organisms. Impairment of stress responses and adrenal gland function, cardiotoxicity, immune system dysfunction, disruption of blood cells and their function, effects on locomotion, and oxidative damage were observed across taxa. This consistency suggests conservation in the mechanisms of action and disease pathogenesis. From a toxicological perspective, a logical progression of impacts was noted: from molecular and cellular effects that manifest as organ dysfunction, to systemic effects that compromise fitness, growth, reproductive potential, and survival. From a clinical perspective, adverse health effects from DWH oil spill exposure formed a suite of signs/symptomatic responses that at the highest doses/concentrations resulted in multi-organ system failure.

Characterization of the differential expressed genes and transcriptomic pathway analysis in the liver of sub-adult red drum (Sciaenops ocellatus) exposed to Deepwater Horizon chemically dispersed oil.

The Deepwater Horizon blowout resulted in the second-largest quantity of chemical dispersants used as a countermeasure for an open water oil spill in the Gulf of Mexico. Of which, the efficacy of dispersant as a mitigation strategy and its toxic effects on aquatic fauna remains controversial. To enhance our understanding of potential sub-lethal effects of exposure to chemically dispersed-oil, sub-adult red drum (Sciaenops ocellatus) were continuously exposed to a Corexit 9500: DWH crude oil chemically enhanced water accommodated fraction (CEWAF) for 3-days and transcriptomic responses were assessed in the liver. Differential expressed gene (DEG) analysis demonstrated that 63 genes were significantly impacted in the CEWAF exposed fish. Of these, 37 were upregulated and 26 downregulated. The upregulated genes were primarily involved in metabolism and oxidative stress, whereas several immune genes were downregulated. Quantitative real-time RT-PCR further confirmed upregulation of cytochrome P450 and glutathione S-transferase, along with downregulation of fucolectin 2 and chemokine C-C motif ligand 20. Ingenuity Pathway Analysis (IPA) predicted 120 pathways significantly altered in the CEWAF exposed red drum. The aryl hydrocarbon receptor pathway was significantly activated, while pathways associated with immune and cellular homeostasis were primarily suppressed. The results of this study indicate that CEWAF exposure significantly affects gene expression and alters signaling of biological pathways important in detoxification, immunity, and normal cellular physiology, which can have potential consequences on organismal fitness.

Characterizing transcriptomic responses of southern flounder (Paralichthys lethostigma) chronically exposed to Deepwater Horizon oiled sediments.

To obtain a deeper understanding of the transcriptomic responses to oil in southern flounder (Paralichthys lethostigma), we performed quantitative PCR and RNA sequencing on liver and gill tissue after a chronic exposure (35 days) to Deepwater Horizon crude oiled sediment and after a 30-day recovery period. We wanted to understand which specific genes are differentially expressed in liver and gill tissues directly after oiled sediment exposure and with the addition of a recovery period. Furthermore, we wanted to examine specific enriched pathways in these two tissues to determine the impact of exposure with and without a recovery period on biological processes (e.g. immune function). Liver and gill tissues were chosen because they represent two distinct organs that are highly important to consider when examining the impacts of oiled sediment exposure. The liver is the classic detoxification organ, while the gill is in direct contact with sediment in benthic fishes. Examination of these two tissues, therefore, generates a broad understanding of the transcriptomic consequences of oil exposure across an organism. Gene expression for interleukin 8 (il8) and interleukin 1B (il1ß) was significantly increased versus control measurements for fish exposed to oiled sediments for 35 days in gill tissue. Hierarchical clustering of gene expression showed that tissue type was the main driver of gene expression (rather than treatment). The inclusion of a 30-day post-exposure recovery period showed a return of il8 and il1ß gene expression in the gill to baseline expression levels. However, the recovery period increased the number of differentially expressed genes and significantly affected canonical pathways in both tissue types. Pathways related to cholesterol biosynthesis were significantly suppressed in oil-exposed flounder with a recovery period, but not in the exposed flounder without a recovery period. At the end of the exposure, 17 pathways were significantly affected in the gill, including thyroid hormone metabolism-related pathways, which were the most influenced. Liver tissue from the recovered fish had the greatest number of enriched pathways for any tissue or time point (187). Cellular and humoral immune response pathways were considerably impacted in the liver after the recovery period, suggesting that the immune system was attempting to respond to potential damage caused from the chronic oil exposure. Our results demonstrate that liver and gill tissues from southern flounder were differentially altered by Deepwater Horizon oiled sediment exposure and that a 30-day recovery period after exposure substantially shifted gene expression and canonical pathway profiles.

Nonlethal Biomarkers of Oxidative Stress in Oiled Sediment Exposed Southern Flounder (Paralichthys lethostigma): Utility for Field-Base Monitoring Exposure and Potential Recovery.

The Deepwater Horizon (DWH) blowout resulted in the deposition of toxic polycyclic aromatic hydrocarbons (PAHs), in the coastal sediments of the Gulf of Mexico. The immediate effects on an ecosystem from an oil spill are clearly recognizable, however the long-term chronic effects and recovery after a spill are still not well understood. Current methodologies for biomonitoring wild populations are invasive and mostly lethal. Here, two potential nonlethal biomonitoring tools for the assessment of PAH toxicity and induced biological alterations in the field, were identified using laboratory-validated methods. In this study, subadult southern flounder (Paralichthys lethostigma) were chronically exposed to DWH surrogate oiled sediments for 35 days; a subset of these exposed flounder were then provided a clean nonexposure period to ascertain the utility of selected biomarkers to monitor recovery post exposure. After chronic exposure, there was an increase in gene expression of cytochrome P450 1A but not glutathione S-transferase. There was also a notable imbalance of oxidants to antioxidants, measured as reduced glutathione, oxidized glutathione, and their ratio in the blood. Evidence of subsequent oxidative damage due to chronic exposure was found through lipid peroxidation and DNA damage assessments of liver, gill, and blood.

Toxicity of Deepwater Horizon source oil and the chemical dispersant, Corexit® 9500, to coral larvae.

Acute catastrophic events can cause significant damage to marine environments in a short time period and may have devastating long-term impacts. In April 2010 the BP-operated Deepwater Horizon (DWH) offshore oil rig exploded, releasing an estimated 760 million liters of crude oil into the Gulf of Mexico. This study examines the potential effects of oil spill exposure on coral larvae of the Florida Keys. Larvae of the brooding coral, Porites astreoides, and the broadcast spawning coral, Montastraea faveolata, were exposed to multiple concentrations of BP Horizon source oil (crude, weathered and WAF), oil in combination with the dispersant Corexit® 9500 (CEWAF), and dispersant alone, and analyzed for behavior, settlement, and survival. Settlement and survival of P. astreoides and M. faveolata larvae decreased with increasing concentrations of WAF, CEWAF and Corexit® 9500, however the degree of the response varied by species and solution. P. astreoides larvae experienced decreased settlement and survival following exposure to 0.62 ppm source oil, while M. faveolata larvae were negatively impacted by 0.65, 1.34 and 1.5 ppm, suggesting that P. astreoides larvae may be more tolerant to WAF exposure than M. faveolata larvae. Exposure to medium and high concentrations of CEWAF (4.28/18.56 and 30.99/35.76 ppm) and dispersant Corexit® 9500 (50 and 100 ppm), significantly decreased larval settlement and survival for both species. Furthermore, exposure to Corexit® 9500 resulted in settlement failure and complete larval mortality after exposure to 50 and 100 ppm for M. faveolata and 100 ppm for P. astreoides. These results indicate that exposure of coral larvae to oil spill related contaminants, particularly the dispersant Corexit® 9500, has the potential to negatively impact coral settlement and survival, thereby affecting the resilience and recovery of coral reefs following exposure to oil and dispersants.

Accumulation and distribution of petroleum hydrocarbons found in mussels (Mytilus galloprovincialis) in the canals of Venice, Italy.

Petroleum contamination was assessed in indigenous and transplanted mussels for three types of environments within the Venice Lagoon and its associated interior canals. Indigenous and transplanted mussels were least impacted by petroleum contamination at open-water stations, but more affected in partially-enclosed areas indicating that physical processes of tidal or wind activities can remove organic contaminants such as petroleum from these areas more effectively. Limited tidal flushing of the interior canals resulted in contaminated locations unable to support indigenous mussels. Clean mussels transplanted to these highly impacted sites accumulated the highest levels of petroleum hydrocarbons. The distribution of polycyclic aromatic hydrocarbons (PAHs) indicated mainly combustion-related activities from sources such as the nearby industrial zones and boat traffic, with some evidence of fresher petroleum inputs. Total hydrocarbons found in all mussels could be related to concentrations found in the surrounding sediments as reported in an earlier study.

Persistence of petroleum hydrocarbon contamination in sediments of the canals in Venice, Italy: 1995 and 1998.

Total hydrocarbon and selected polycyclic aromatic hydrocarbon (PAH) levels were examined in sediments collected from known problematic areas in the canals of Venice, Italy, in 1995 and 1998. Hydrocarbon concentrations were greatest in the interior canals, moderate in the partially enclosed locations and lowest in the open-water sites. Total hydrocarbon and PAH concentrations declined from 1995 to 1998. Ancillary data suggest that this decline may have been in response to the elimination of many industrial activities in the lagoon and to initiating an aggressive canal dredging program. The distributions of individual components were generally similar both years regardless of the total PAH concentration or the location of sample collected. PAH alkyl homolog distributions suggest that atmospheric deposition of petrochemical combustion products is the main source of PAHs to Venice's sediments. In some cases however, the presence of low levels of the two-ring naphthalene homologous series indicate additional low-level inputs of fresh oil.