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.

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.

Acute exposure to oil induces age and species-specific transcriptional responses in embryo-larval estuarine fish.

Because oil spills frequently occur in coastal regions that serve as spawning habitat, characterizing the effects of oil in estuarine fish carries both economic and environmental importance. There is a breadth of research investigating the effects of crude oil on fish, however few studies have addressed how transcriptional responses to oil change throughout development or how these responses might be conserved across taxa. To investigate these effects, we performed RNA-seq and pathway analysis following oil exposure 1) in a single estuarine species (Cyprinodon variegatus) at three developmental time points (embryos, yolk-sack larvae, free-feeding larvae), and 2) in two ecologically similar species (C. variegatus and Fundulus grandis), immediately post-hatch (yolk-sack stage). Our results indicate that C. variegatus embryos mount a diminished transcriptional response to oil compared to later stages, and that few transcriptional responses are conserved throughout development. Pathway analysis of larval C. variegatus revealed dysregulation of similar biological processes at later larval stages, including alteration of cholesterol biosynthesis pathways, cardiac development processes, and immune functions. Our cross-species comparison showed that F. grandis exhibited a reduced transcriptional response compared to C. variegatus. Pathway analysis revealed that the two species shared similar immune and cardiac responses, however pathways related to cholesterol biosynthesis exhibited a divergent response as they were activated in C. variegatus but inhibited in F. grandis. Our results suggest that examination of larval stages may provide a more sensitive estimate of oil-impacts than examination of embryos, and challenge assumptions that ecologically comparable species respond to oil similarly.

Parental exposure to Deepwater Horizon oil in different environmental scenarios alters development of sheepshead minnow (Cyprinodon variegatus) offspring.

The explosion of the Deepwater Horizon (DWH) oil exploration platform on April 20, 2010 began a catastrophic leak of approximately 640 million liters crude oil into the northern Gulf of Mexico (GOM), affecting more than 2100 km of coastline, including wetlands and estuaries that provide habitat and nursery for many aquatic species. Estuaries of the GOM are dynamic environments, with constant fluctuations in salinity and dissolved oxygen, including large hypoxic zones during summer months. Spawning fish in northern GOM estuaries following the DWH incident were at significant risk of oil exposure, and adverse environmental conditions at the time of exposure, such as hypoxia and low salinity, could have exacerbated developmental effects in the offspring. The present study investigated the effects of F0 parental oil exposure in different environmental scenarios on development of F1 sheepshead minnow (SHM) offspring. Adult SHM were exposed to the high-energy water accommodated fraction (HEWAF) of crude oil in three environmental scenarios: normoxic (NORM), hypoxic (HYP), and hypoxic with low salinity (HYP-LS). Parental HEWAF exposure in the NORM scenario resulted in developmental effects in F1 offspring, including altered heart rate, decreased length at hatch, and impaired prey capture. Co-exposure of F0 SHM to HEWAF and adverse environmental conditions altered HEWAF effects on F1 heart rate, hatch rate, prey capture, and survival. Time to hatch was not significantly impacted by parental HEWAF in any environmental scenario. The present study demonstrates that parental exposure to HEWAF results in developmental changes in F1 embryos, and co-exposure to adverse environmental conditions altered the effects for several developmental endpoints. These data suggest that SHM exposed to oil in estuaries experiencing hypoxia or low salinity may produce offspring with worsened outcomes. These developmental effects, in addition to previously reported reproductive effects in adult fish, could lead to long-term population level impacts for SHM.

The combined effects of salinity, hypoxia, and oil exposure on survival and gene expression in developing sheepshead minnows, Cyprinodon variegatus.

The 2010 Deepwater Horizon oil spill released approximately 780 million liters of crude oil contaminating coastal habitats from Texas to Florida which are important habitats for many fish species during early life stages. These diverse habitats are also prone to rapid fluctuations in water quality, such as dissolved oxygen concentration and salinity. The consequence of combined exposure to crude oil and suboptimal environmental conditions during early life stage development of fish is still largely unknown. The objective of this project was to investigate the impacts of exposure to crude oil in combination with varying environmental stressors on developing Cyprinodon variegatus survival, growth, and gene expression. Three life stages (embryonic, post-hatch, and post-larval) were exposed to four nominal concentrations (6.25%, 12.5%, 50% and 100% with actual polycyclic aromatic hydrocarbon (PAH) concentrations ranging from 0 to 512 µg/L) of high energy water accommodated fractions (HEWAF) under different oxic (2.0 or >5.0 mg/L) and salinity (10 or 30 ppt) regimes at 30 °C for 48 h. We found that the post-larval developmental stage was the most sensitive to oil toxicity. Median lethal concentrations during the post-larval exposures followed a treatment-dependent pattern with the highest mortality observed under hypoxic-high salinity conditions (64.55 µg/L). Real-time PCR analysis identified down regulation of target genes, encoding cytochrome P450-1α (cyp1a1), erythropoietin (epo), and the aryl hydrocarbon receptor nuclear translocator (arnt1) only when oil exposure occurred under hypoxic-high salinity conditions in treatments with PAH concentrations greater than 226 µg/L. The target genes measured in this experiment are involved in the aryl hydrocarbon receptor signaling pathway which modulates metabolism of PAHs (a major component of crude oil), and the hypoxia inducible 1-α signaling pathway which is responsible for resilience to hypoxic stress, and it is known that disruption of these pathways can lead to an array of acute and chronic effects. Our results indicated that sheepshead minnow are most sensitive to oil exposure during the post-larval developmental stage. Survival data from this age-stage also indicate that oil toxicity response is exacerbated in hypoxic and high salinity environments. The increased mortality observed during the post-larval developmental stage might be attributed to the suppression of the aryl hydrocarbon receptor signaling and the hypoxia inducible 1-α signaling pathways which is evident in by the down-regulated expression of cyp1a1, epo, and arnt1. These findings provide more information about interactions between oil and abiotic factors which enable us to make better assumptions of the ecological impacts of DWH on coastal estuaries.

Combined effects of salinity, temperature, hypoxia, and Deepwater Horizon oil on Fundulus grandis larvae.

Oil spills have polluted the marine environment for decades and continue to be a major source of polycyclic aromatic hydrocarbons (PAHs) to marine ecosystems around the globe, for example during the 2010 Deepwater Horizon spill. Although the toxicity of PAHs to fish has been well studied, their effects combined with abiotic stressors are poorly understood. The goal of this study was to describe the combined impacts of crude oil and environmental stressors on fish larvae, a sensitive life stage. Gulf killifish (Fundulus grandis) larvae (<24 h post-hatch) were exposed for 48 h to high energy water accommodated fractions (HEWAF; total PAHs 0-125 ppb) of Macondo oil from the Deepwater Horizon spill under different combinations of environmental conditions (dissolved oxygen 2, 6 ppm; temperature 20, 25, 30 °C; salinity 3, 10, 30 ppt). Even under optimal environmental conditions (25 °C, 10 ppt, 6 ppm) larval survival and development were negatively affected by PAHs, starting with the lowest concentration tested (∼15 ppb). Hypoxia and high temperature each increased the adverse effects of HEWAF on development and mortality. In contrast, salinity had little effect on any of the endpoints measured. Importantly, expression of the detoxifying gene cyp1a was highly induced in PAH-exposed larvae under normoxic conditions, but not under hypoxic conditions, potentially explaining the enhanced toxicity observed under hypoxia. This work highlights the importance of considering how suboptimal environmental conditions can exacerbate the effects of pollution on fish early life stages.

Hypoxia and reduced salinity exacerbate the effects of oil exposure on sheepshead minnow (Cyprinodon variegatus) reproduction.

Estuaries of the northern Gulf of Mexico are dynamic environments, with fluctuations in salinity and dissolved oxygen, including areas of seasonal hypoxia. Fish that reside and reproduce in these estuaries, including sheepshead minnow (Cyprinodon variegatus; SHM), were at significant risk of oil exposure following the Deepwater Horizon oil spill. It is poorly understood how differences in environmental conditions during oil exposure impact its toxicity. The present study investigated the effects of crude oil high-energy water accommodated fraction (HEWAF) on SHM reproduction in three environmental scenarios (normoxic, hypoxic, and hypoxic with low salinity) to determine if differences in salinity (brackish vs low salinity) and dissolved oxygen (normoxia vs hypoxia) could exacerbate the effects of HEWAF-derived polycyclic aromatic hydrocarbons (PAHs). We observed that HEWAF exposure significantly increased liver somatic index of SHM compared to control, but this effect was not exacerbated by hypoxia or low salinity. HEWAF exposure also significantly decreased egg production and egg fertilization rate, but only in the hypoxic and hypoxic with low salinity scenarios. A significant correlation existed between body burdens of PAHs and reproductive endpoints, providing substantial evidence that oil exposure reduced reproductive capacity in SHM, across a range of environmental conditions. These data suggest that oil spill risk assessments that fail to consider other environmental stressors (i.e. hypoxia and salinity) may be underestimating risk.

Transgenerational effects of polycyclic aromatic hydrocarbon exposure on sheepshead minnows (Cyprinodon variegatus).

The Deepwater Horizon oil spill resulted in the release of over 640 million L of crude oil into the Gulf of Mexico, affecting over 2000 km of shoreline, including estuaries that serve as important habitats and nurseries for aquatic species. Cyprinodon variegatus (sheepshead minnow) are small-bodied fish that inhabit northern Gulf of Mexico estuaries, are easily adaptable to laboratory conditions, and are commonly used in toxicological assessment studies. The purpose of the present study was to determine the somatic, reproductive, and developmental effects of an environmentally relevant polycyclic aromatic hydrocarbon (PAH) mixture, the oil high-energy water accommodated fraction (HEWAF), on experimentally exposed sheepshead minnow (F0 ) as well as 2 generations of offspring (F1 and F2 ) without additional exposure. The F0 generation exposed to HEWAF had increased liver somatic indices, altered egg production, and decreased fertilization. Several developmental endpoints in the F1 were altered by F0 HEWAF exposure. As adults, low HEWAF-exposed F1 females demonstrated decreased weight and length. Both the F1 and F2 generations derived from high HEWAF-exposed F0 had deficits in prey capture compared to control F1 and F2 , respectively. Correlations between endpoints and tissue PAHs provide evidence that the physiological effects observed were associated with hydrocarbon exposure. These data demonstrate that PAHs were capable of causing physiological changes in exposed adult sheepshead minnow and transgenerational effects in unexposed offspring, both of which could have population-level consequences. Environ Toxicol Chem 2019;38:638-649. © 2018 SETAC.

Combined effects of Deepwater Horizon crude oil and environmental stressors on Fundulus grandis embryos.

In the present study, we examined how sensitivity to oil changes in combination with environmental stressors in Fundulus grandis embryos. We exposed embryos (<24 h post fertilization) to a range of high-energy water accommodated fraction (HEWAF) concentrations (0-50 parts per billion [ppb] total polycyclic aromatic hydrocarbons [PAHs]) made from Macondo crude oil in conjunction with various environmental conditions (temperature: 20 and 30 °C; salinity: 3, 7, and 30 practical salinity units [PSU]; and dissolved oxygen: 2 and 6 mg/L). Endpoints included mortality, hatching rates, and expression of cytochrome p450 1a and 1c (cyp1a, cyp1c) in hatched larvae. There was 100% mortality for all fish under the 2 parts per million (ppm) dissolved oxygen regimes. For the 6 mg/L dissolved oxygen treatments, mortality and median lethal time (LT50) were generally higher in the 30 °C treatments versus the 20 °C treatments. Oil increased mortality in fish exposed to the highest concentration in the 20-3-6 (°C-PSU-mg/L), 25-7-6, and 30-30-6 conditions. Hatching was driven by environmental conditions, with oil exposure having a significant impact on hatching in only the 25-7-6 and 30-30-6 groups at the greatest HEWAF exposure. Expression of cyp1a was up-regulated in most treatment groups versus the controls, with cyp1c expression exhibiting a similar pattern. These data suggest interactive effects among temperature, salinity, and PAHs, highlighting a need to further assess the effects of oil exposure under various environmental conditions. Environ Toxicol Chem 2018;37:1916-1925. © 2018 SETAC.

Exposure to Deepwater Horizon oil and Corexit 9500 at low concentrations induces transcriptional changes and alters immune transcriptional pathways in sheepshead minnows.

The 2010 Deepwater Horizon (DWH) oil spill caused the release of 4.9 million barrels of crude oil into the Gulf of Mexico, followed by the application of 2.9 million L of the dispersant, Corexit™ to mitigate the spread of oil. The spill resulted in substantial shoreline oiling, potentially exposing coastal organisms to polyaromatic hydrocarbon (PAH) and dispersant contaminants. To investigate molecular effects in fish following exposure to environmentally relevant concentrations of DWH oil and dispersants, we exposed adult sheepshead minnows (Cyprinodon variegatus) to two concentrations of high-energy water-accommodated fraction (HEWAF), chemically enhanced water-accommodated fraction (CEWAF) or Corexit 9500™ for 7 and 14days. Resulting changes in hepatic gene expression were measured using 8×15K microarrays. Analytical chemistry confirmed PAH concentrations in HEWAF and CEWAF treatments were low (ranging from 0.26 to 5.98µg/L), and likely representative of post-spill environmental concentrations. We observed significant changes to gene expression in all treatments (relative to controls), with Corexit and CEWAF having a greater effect on expression patterns in the liver than HEWAF treatments. Sub-network enrichment analysis of biological pathways revealed that the greatest number of altered pathways in high dose HEWAF and CEWAF treatments occurred following a 7-day exposure. Pathways related to immunity comprised the majority of pathways affected in each treatment, followed by pathways related to blood and circulation processes. Our results indicate that oil composition, concentration, and exposure duration all affect molecular responses in exposed fish, and suggest that low-concentration exposures may result in sub-lethal adverse effects.

Crude oil impairs immune function and increases susceptibility to pathogenic bacteria in southern flounder.

Exposure to crude oil or its individual constituents can have detrimental impacts on fish species, including impairment of the immune response. Increased observations of skin lesions in northern Gulf of Mexico fish during the 2010 Deepwater Horizon oil spill indicated the possibility of oil-induced immunocompromisation resulting in bacterial or viral infection. This study used a full factorial design of oil exposure and bacterial challenge to examine how oil exposure impairs southern flounder (Paralichthys lethostigma) immune function and increases susceptibility to the bacteria Vibrio anguillarum, a causative agent of vibriosis. Fish exposed to oil prior to bacterial challenge exhibited 94.4% mortality within 48 hours of bacterial exposure. Flounder challenged with V. anguillarum without prior oil exposure had <10% mortality. Exposure resulted in taxonomically distinct gill and intestine bacterial communities. Mortality strongly correlated with V. anguillarum levels, where it comprised a significantly higher percentage of the microbiome in Oil/Pathogen challenged fish and was nearly non-existent in the No Oil/Pathogen challenged fish bacterial community. Elevated V. anguillarum levels were a direct result of oil exposure-induced immunosuppression. Oil-exposure reduced expression of immunoglobulin M, the major systemic fish antibody, and resulted in an overall downregulation in transcriptome response, particularly in genes related to immune function, response to stimulus and hemostasis. Ultimately, sediment-borne oil exposure impairs immune function, leading to increased incidences of bacterial infections. This type of sediment-borne exposure may result in long-term marine ecosystem effects, as oil-bound sediment in the northern Gulf of Mexico will likely remain a contamination source for years to come.

Responses of juvenile southern flounder exposed to Deepwater Horizon oil-contaminated sediments.

The Deepwater Horizon oil spill released millions of barrels of crude oil into the northern Gulf of Mexico, much of which remains associated with sediments and can have continuing impacts on biota. Juvenile southern flounder (Paralichthys lethostigma) were exposed for 28 d in the laboratory under controlled conditions to reference and Deepwater Horizon oil-contaminated sediments collected from coastal Louisiana to assess the impacts on an ecologically and commercially important benthic fish. The measured polycyclic aromatic hydrocarbon (PAH) concentrations in the sediments ranged from 0.25 mg/kg to 3940 mg/kg suite of 50 PAH analytes (tPAH50). Mortality increased with both concentration and duration of exposure. Exposed flounder length and weight was lower compared to controls after 28 d of exposure to the sediments with the highest PAH concentration, but condition factor was significantly higher in these fish compared with all other treatments. Histopathological analyses showed increased occurrence of gill abnormalities, including telangiectasis, epithelial proliferation, and fused lamellae in flounder exposed to sediments with the highest tPAH50 concentrations. In addition, hepatic vascular congestion and macrovesicular vacuolation were observed in flounder exposed to the more contaminated sediments. These data suggest that chronic exposure to field collected oil-contaminated sediments results in a variety of sublethal impacts to a benthic fish, with implications for long-term recovery from oil spills. Environ Toxicol Chem 2017;36:1067-1076. © 2016 SETAC.

Simultaneous exposure to chronic hypoxia and dissolved polycyclic aromatic hydrocarbons results in reduced egg production and larval survival in the sheepshead minnow (Cyprinodon variegatus).

Estuarine fish in the northern Gulf of Mexico are exposed annually to hypoxic conditions. In addition to hypoxia, fish located throughout the northern Gulf of Mexico were potentially exposed to oil released during the Deepwater Horizon incident. Therefore, the interaction between oil exposure and hypoxia is worth investigating. To examine this interaction, the authors exposed adult and larval sheepshead minnow (Cyprinodon variegatus) to crude or dispersed oil under both normoxic and hypoxic conditions. The authors examined total egg production, egg hatching success, and larval survival post hatch. The authors' results indicate that co-exposure to crude or dispersed oil and hypoxia resulted in a significant decrease in egg production, as well as a significant decrease in both egg hatch success and larval survival post hatch. The significant impact on reproductive success following crude or dispersed oil and hypoxia exposure indicates the importance of including environmental parameters such as hypoxia when evaluating the impact of an oil spill.

Photo-induced toxicity of Deepwater Horizon slick oil to blue crab (Callinectes sapidus) larvae.

The 2010 Deepwater Horizon oil spill resulted in the accidental release of approximately 700 million L of crude oil into the Gulf of Mexico. Photo-induced toxicity after co-exposure to ultraviolet (UV) radiation is 1 mechanism by which polycyclic aromatic hydrocarbons (PAHs) from oil spills may exert toxicity. Blue crab are an important commercial and ecological resource in the Gulf of Mexico, and their largely transparent larvae may make them sensitive to PAH photo-induced toxicity. The goal of the present study was to examine the sensitivity of early lifestage blue crab (Callinectes sapidus) zoea to slick oil collected during the Deepwater Horizon spill. Blue crab zoea were exposed to 1 of several dilutions of water accommodated fractions from 1 of 2 sources of oil and gradations of natural sunlight in a factorial design. Two 7-h solar exposures were carried out with a recovery period (dark) in between. Survival was found to be UV- and PAH-dependent. Toxicity was observed within the range of surface PAH concentrations reported in the Gulf of Mexico during the Deepwater Horizon spill. These findings indicate that early lifestage blue crab are sensitive to photo-induced toxicity from Deepwater Horizon slick oil.

A multiple endpoint analysis of the effects of chronic exposure to sediment contaminated with Deepwater Horizon oil on juvenile Southern flounder and their associated microbiomes.

Exposure to oiled sediments can negatively impact the health of fish species. Here, we examine the effects of chronic exposure of juvenile southern flounder, Paralichthys lethostigma, to a sediment-oil mixture. Oil:sediment mixtures are persistent over time and can become bioavailable following sediment perturbation or resuspension. Juvenile flounder were exposed for 32 days under controlled laboratory conditions to five concentrations of naturally weathered Macondo MC252 oil mixed into uncontaminated, field-collected sediments. The percent composition of individual polycyclic aromatic hydrocarbons (PAHs) of the weathered oil did not change after mixing with the sediment. Spiked exposure sediments contained 0.04-395mg/kg tPAH50 (sum of 50 individual PAH concentration measurements). Mortality increased with both exposure duration and concentration of sediment-associated PAHs, and flounder exposed to concentrations above 8mg/kg tPAH50 showed significantly reduced growth over the course of the experiment. Evident histopathologic changes were observed in liver and gill tissues of fish exposed to more than 8mg/kg tPAH50. All fish at these concentrations showed hepatic intravascular congestion, macrovesicular hepatic vacoulation, telangiectasia of secondary lamellae, and lamellar epithelial proliferation in gill tissues. Dose-dependent upregulation of Cyp1a expression in liver tissues was observed. Taxonomic analysis of gill and intestinal commensal bacterial assemblages showed that exposure to oiled sediments led to distinct shifts in commensal bacterial population structures. These data show that chronic exposure to environmentally-relevant concentrations of oiled sediments produces adverse effects in flounder at multiple biological levels.

Gene expression and growth as indicators of effects of the BP Deepwater Horizon oil spill on spotted seatrout (Cynoscion nebulosus).

The BP Deepwater Horizon oil spill has great potential to negatively affect estuarine fish populations. In order to assess possible impacts of this event, a series of sublethal lab experiments were performed, using the economically and ecologically important species spotted seatrout (Cynoscion nebulosus). Larval and juvenile spotted seatrout were exposed to sublethal concentrations of high energy water accommodated fraction (HEWAF), chemically enhanced water accommodated fraction (CEWAF), or dispersant alone in an acute exposure. Response to exposure was evaluated with quantative polymerase chain reaction (qPCR) to examine expression of cytochrome P-4501A (CYP1A). Growth of larvae and juveniles over the duration of the experiment was measured as an index of physiological response. Our data showed that the different life stages respond differently to crude and dispersed oil, with larval spotted seatrout affected most by CEWAF, while juvenile spotted seatrout were affected to a greater extent by HEWAF. In both cases, the treatment with the highest CYP1A levels resulted in the greatest reductions in growth.