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The unprecedented size of the deepwater horizon oil spill and scope of the subsequent response elicited intense and sustained interest in microbial responses to oiling, especially in salt marshes, which have featured prominently in debates about best practices to prevent and remediate oiling of vulnerable ecosystems. A number of studies have examined salt marsh soil microbial communities following the spill, but most have primarily concentrated on prokaryotes. The extent to which oiling elicited shifts in fungal diversity and community composition remains unclear. Here we present spatial and temporal comparisons of salt marsh soil fungal communities at two southern Louisiana salt marshes with contrasting oiling histories. We profiled fungal communities in 2013 alongside corresponding measurements of polycyclic aromatic hydrocarbons to assess whether and how responses to oiling are distinguishable from natural heterogeneity. Analyses based on high-resolution unbiased spatial sampling demonstrated that fungal communities did not align with shoreline classification of oiling less than three years after initial oiling, despite observable differences in oil residues and secondary oiling. Notably, extensive sampling allowed delineation of benchmark sampling thresholds and illustrated the value of using ranked differentials of relative abundance to characterize soil fungal communities. Our findings highlight the need for combining high-resolution sampling with judgment-based and systematic sampling approaches to accurately capture responses of salt marsh soil fungal community to oiling.

The 2010 Deepwater Horizon disaster remains one of the largest oil spills in history. This event caused significant damage to coastal ecosystems, the full extent of which has yet to be fully determined. Crude oil contains toxic heavy metals and substances such as polycyclic aromatic hydrocarbons that are detrimental to some microbial species and may be used as food and energy resources by others. As a result, oil spills have the potential to cause significant shifts in microbial communities. This study assessed the impact of oil contamination on the function of endophytic microbial communities associated with saltmarsh cordgrass (Spartina alterniflora). Soil samples were collected from two locations in coastal Louisiana, USA: one severely affected by the Deepwater Horizon oil spill and one relatively unaffected location. Spartina alterniflora seedlings were grown in both soil samples in greenhouses, and GeoChip 5.0 was used to evaluate the endophytic microbial metatranscriptome shifts in response to host plant oil exposure. Oil exposure was associated with significant shifts in microbial gene expression in functional categories related to carbon cycling, virulence, metal homeostasis, organic remediation, and phosphorus utilization. Notably, significant increases in expression were observed in genes related to metal detoxification with the exception of chromium, and both significant increases and decreases in expression were observed in functional gene subcategories related to hydrocarbon metabolism. These findings show that host oil exposure elicits multiple changes in gene expression from their endophytic microbial communities, producing effects that may potentially impact host plant fitness.

Storm-driven transport of MC252 oil, sand and shell aggregates was studied on a low-relief coastal headland beach in Louisiana, USA including measurement of alkylated PAHs and Illumina sequencing of intra-aggregate microbial populations. Weathering ratios, constructed from alkylated PAH data, were used to assess loss of 3-ring phenanthrenes and dibenzothiophenes relative to 4-ring chrysenes. Specific aggregate types showed relatively little weathering of 3-ring PAHs referenced to oil sampled near the Macondo wellhead with the exception of certain SRBs sampled from the supratidal environment and samples from deposition areas north of beach. Aggregates mobilized by these storm-driven washover events contains diverse microbial populations dominated by the class Gammaproteobacteria including PAH-degrading genera such as Halomonas, Marinobacter and Idiomarina. Geochemical assessment of porewater in deposition areas, weathering observations, and microbial data suggest that storm remobilization can contribute to susceptibility of PAHs to biodegradation by moving oil to beach microenvironments with more favorable characteristics. (149).

Complementary microbial and geochemical assessment techniques investigated the biodegradation of PAHs and alkanes in salt marshes impacted by crude oil following the Macondo spill. Contamination was observed in the top 10 cm of the marsh profile based on PAH analysis and measurement of the δ13C signature of impacted marsh soils. Measurement of evolution of 13C depleted CO2 indicated mineralization of crude oil ranging from 2.7–12.1 mg CO2-C/m2-hr. Changes in weathering ratios of alkylated phenanthrenes and dibenzothiophenes indicated loss of these 3-ring PAHs consistent with biodegradation. A diverse microbial population was observed at both locations dominated by Gammaproteobacteria and including known hydrocarbon degraders such as Marinobacter and Alcanivorax. There was shared richness between sites and across seasons but results suggested substantial turnover of phylotypes in space and time. Biodegradation of alkanes and alkylated PAHs occurred when oxygen was provided in laboratory reactors but not in the absence of oxygen.

Laboratory experiments investigated oxygen dynamics in buried oiled sands sampled from areas impacted by the Macondo spill. Measured oxygen fluxes in oil deposits that were permeable to tidal water ranged from 10−3 to 10−4 μmol/cm2-sec, orders of magnitude higher than fluxes in non-permeable deposits (10−6 to 10−7 μmol/cm2-sec). Oxygen dynamics were well described by 1-d models that represent increased oxygen consumption in oiled sands. Experiments demonstrated that when oxygen is present and the oil deposit is permeable to tidal water, biodegradation of alkylated phenanthrenes and dibenzothiophenes proceeded over time scales (i.e., weeks) to have a significant impact on the mass and quality of buried oil. For this biodegradation process to proceed, two independent conditions must be met, a source of oxygenated water has to be present (e.g., tidal flushing in the intertidal zone) and the oiled deposit has to be permeable to water (i.e., pores cannot be completely saturated with oil).

The plant microbiome, composed of diverse interacting microorganisms, is thought to undergird host integrity and well-being. Though it is well understood that environmental perturbations like oil pollution can alter the diversity and composition of microbiomes, remarkably little is known about how disturbance alters plant-fungal associations. Using Next-Generation sequencing of the 18S rDNA internal transcribed spacer (ITS1) region, we examined outcomes of enduring oil exposure on aboveground leaf and belowground endophytic root and rhizosphere fungal communities of Spartina alterniflora, a highly valued ecosystem engineer in southeastern Louisiana marshes affected by the 2010 Deepwater Horizon accident. We found that aboveground foliar fungal communities exhibited site-dependent compositional turnover with consequent loss in diversity according to oiling history. Rhizosphere soil communities also exhibited shifts in community composition associated with oiling history, whereas root endophytic communities did not. Oiling did not increase or decrease similarities among aboveground and belowground communities within an individual host, indicating that host plant characteristics exert stronger control than external factors on fungal community composition. These results show that fungal community responses to oiling vary within tissues of the same host plant, and that differences in the local environment, or alternatively, site-specific differences in residual oil constrain the magnitude of exposure responses. Our study offers novel perspectives on how environmental contaminants and perturbations can influence plant microbiomes, highlighting the importance of assessing long-term ecological outcomes of oil pollution to better understand how shifts in microbial communities influence plant performance and ecosystem function. Our findings are relevant to coastal management programs tasked with responding to oil spills and increasing pressures arising from intensifying development and climate change. Understanding how modification of plant-microbiome associations influences plant performance, particularly of ecosystem engineers like S. alterniflora, can help guide efforts to protect and restore at-risk coastal ecosystems.

The distribution and characteristics of MC252 oil:sand aggregates, termed surface residue balls (SRBs), were measured on the supratidal beach environment of oil-impacted Fourchon Beach in Louisiana (USA). Probability distributions of 4 variables, surface coverage (%), size of SRBs (mm2 of projected area), mass of SRBs per m2 (g/m2), and concentrations of polycyclic aromatic hydrocarbons (PAHs) and n-alkanes in the SRBs (mg of crude oil component per kg of SRB) were determined using parametric and nonparametric statistical techniques. Surface coverage of SRBs, an operational remedial standard for the beach surface, was a gamma-distributed variable ranging from 0.01% to 8.1%. The SRB sizes had a mean of 90.7 mm2 but fit no probability distribution, and a nonparametric ranking was used to describe the size distributions. Concentrations of total PAHs ranged from 2.5 mg/kg to 126 mg/kg of SRB. Individual PAH concentration distributions, consisting primarily of alkylated phenanthrenes, dibenzothiophenes, and chrysenes, did not consistently fit a parametric distribution. Surface coverage was correlated with an oil mass per unit area but with a substantial error at lower coverage (i.e.

Buried and surface MC252 oil from the intertidal on a coastal headland beach was sampled using a randomized block methodology. Weathering indices contrasting alkylated 3- (phenanthrenes) and 4-ring (chrysene) PAH concentrations were computed based on analyses of these samples. Buried oil was detected at a frequency of 18% and, of those samples, 29% had indices indistinguishable from oil sampled near the wellhead indicating that PAH weathering was not occurring. These samples with persistent PAHs were associated with one or more conditions in the beach profile: located at or near the depth of the water table, nearest to the shoreline, or within a thicker oil mat deposit. Surface oil samples, consisting of oil:sand aggregates recently washed in by the surf, had a higher percentage (81%) of oil with weathering indices indicating persistence. These observations, coupled with measurements of biogeochemical analyses on the beach from other studies, suggest that insufficient oxygen in the nearshore environment of these coastal headlands creates conditions for PAH persistence over time frames of years to decades.

Unique oil:sand aggregates, termed surface residue balls (SRBs), were formed on coastal headland beaches along the northern Gulf of Mexico as emulsified MC252 crude oil mixed with sand following the Deepwater Horizon spill event. The objective of this study is to assess the biodegradation potential of crude oil components in these aggregates using multiple lines of evidence on a heavily-impacted coastal headland beach in Louisiana, USA. SRBs were sampled over a 19-month period on the supratidal beach environment with reasonable control over and knowledge of the residence time of the aggregates on the beach surface. Polycyclic aromatic hydrocarbons (PAHs) and alkane concentration ratios were measured including PAH/C30-hopane, C2/C3 phenanthrenes, C2/C3 dibenzothiophenes and alkane/C30-hopane and demonstrated that biodegradation was occurring in SRBs in the supratidal. These biodegradation reactions occurred over time frames relevant to the coastal processes moving SRBs off the beach. In contrast, submerged oil mat samples from the intertidal did not demonstrate chemical changes consistent with biodegradation. Review and analysis of additional biogeochemical parameters suggested the existence of a moisture and nutrient-limited biodegradation regime on the supratidal beach environment. At this location, SRBs possess moisture contents

The distribution and characteristics of MC252 oil:sand aggregates, termed surface residue balls (SRBs), were measured on the supratidal beach environment of oil-impacted Fourchon Beach in Louisiana (USA). Probability distributions of 4 variables, surface coverage (%), size of SRBs (mm(2) of projected area), mass of SRBs per m(2) (g/m(2)), and concentrations of polycyclic aromatic hydrocarbons (PAHs) and n-alkanes in the SRBs (mg of crude oil component per kg of SRB) were determined using parametric and nonparametric statistical techniques. Surface coverage of SRBs, an operational remedial standard for the beach surface, was a gamma-distributed variable ranging from 0.01% to 8.1%. The SRB sizes had a mean of 90.7 mm(2) but fit no probability distribution, and a nonparametric ranking was used to describe the size distributions. Concentrations of total PAHs ranged from 2.5 mg/kg to 126 mg/kg of SRB. Individual PAH concentration distributions, consisting primarily of alkylated phenanthrenes, dibenzothiophenes, and chrysenes, did not consistently fit a parametric distribution. Surface coverage was correlated with an oil mass per unit area but with a substantial error at lower coverage (i.e.,2%). These data provide probabilistic risk assessors with the ability to specify uncertainty in PAH concentration, exposure frequency, and ingestion rate, based on SRB characteristics for the dominant oil form on beaches along the US Gulf Coast.

MC252 oil:sand aggregates, termed surface residue balls (SRBs), were sampled for physical, chemical and microbial characteristics from different tidal zones on a coastal headland beach in Louisiana, USA. Supratidal SRBs were smaller, had low moisture content, and salinities that were