Hydrocarbons in Deep-Sea Sediments following the 2010 Deepwater Horizon Blowout in the Northeast Gulf of Mexico

The Deepwater Horizon (DWH) spill released 4.9 million barrels of oil into the Gulf of Mexico (GoM) over 87 days. Sediment and water sampling efforts were concentrated SW of the DWH and in coastal areas. Here we present geochemistry data from sediment cores collected in the aftermath of the DWH event from 1000 – 1500 m water depth in the DeSoto Canyon, NE of the DWH wellhead. Cores were analyzed at high-resolution (at 2 mm and 5 mm intervals) in order to evaluate the concentration, composition and input of hydrocarbons to the seafloor. Specifically, we analyzed total organic carbon (TOC), aliphatic, polycyclic aromatic hydrocarbon (PAHs), and biomarker (hopanes, steranes, diasteranes) compounds to elucidate possible sources and transport pathways for deposition of hydrocarbons. Results showed higher hydrocarbon concentrations during 2010-2011 compared to years prior to 2010. Hydrocarbon inputs in 2010-2011 were composed of a mixture of sources including terrestrial, planktonic, and weathered oil. Our results suggest that after the DWH event, both soluble and highly insoluble hydrocarbons were deposited at enhanced rates in the deep-sea. We proposed two distinct transport pathways of hydrocarbon deposition: 1) sinking of oil-particle aggregates (hydrocarbon-contaminated marine snow and/or suspended particulate material), and 2) advective transport and direct contact of the deep plume with the continental slope surface sediments between 1000-1200 m. Our findings underline the complexity of the depositional event observed in the aftermath of the DWH event in terms of multiple sources, variable concentrations, and spatial (depth-related) variability in the DeSoto Canyon, NE of the DWH wellhead.     

Deep-Sea Benthic Footprint of the Deepwater Horizon Blowout

The Deepwater Horizon (DWH) accident in the northern Gulf of Mexico occurred on April 20, 2010 at a water depth of 1525 meters, and a deep-sea plume was detected within one month. Oil contacted and persisted in parts of the bottom of the deep-sea in the Gulf of Mexico. As part of the response to the accident, monitoring cruises were deployed in fall 2010 to measure potential impacts on the two main soft-bottom benthic invertebrate groups: macrofauna and meiofauna. Sediment was collected using a multicorer so that samples for chemical, physical and biological analyses could be taken simultaneously and analyzed using multivariate methods. The footprint of the oil spill was identified by creating a new variable with principal components analysis where the first factor was indicative of the oil spill impacts and this new variable mapped in a geographic information system to identify the area of the oil spill footprint. The most severe relative reduction of faunal abundance and diversity extended to 3 km from the wellhead in all directions covering an area about 24 km². Moderate impacts were observed up to 17 km towards the southwest and 8.5 km towards the northeast of the wellhead, covering an area 148 km². Benthic effects were correlated to total petroleum hydrocarbon, polycyclic aromatic hydrocarbons and barium concentrations, and distance to the wellhead; but not distance to hydrocarbon seeps. Thus, benthic effects are more likely due to the oil spill, and not natural hydrocarbon seepage. Recovery rates in the deep sea are likely to be slow, on the order of decades or longer.     

Oiling rates and condition indices of shorebirds on the northern Gulf of Mexico following the Deepwater Horizon oil spill

The coastline of the Gulf of Mexico in the United States is an important wintering and stopover region for migratory shorebirds. The Deepwater Horizon oil spill (April–August 2010) impacted more than 1700 km of this coastline and could potentially affect shorebirds through long‐term exposure to toxins, degraded habitats, and altered food chains. We investigated the exposure to Deepwater Horizon oil of seven species of shorebirds that winter or stopover along the northern Gulf of Mexico. From October 2010 to May 2012, we captured and banded 691 shorebirds at six sites that experienced varying levels of oil contamination. Of birds sampled, 22 were lightly oiled, with species that forage on the coast having higher rates of oiling than those that forage in more estuarine habitats. Although only 8.6% of birds captured from October 2010 to May 2011 and 0.6% of the birds captured from August 2011 to June 2012 showed signs of oiling, an unknown, but potentially larger, number of shorebirds were likely exposed to indirect effects of the spill, such as decreased foraging time due to oiling of sites or disturbance from cleanup activities. Fuel stores and fattening rates of Dunlins (Calidris alpina) during spring migration, as measured using plasma metabolites, were not influenced by site oiling level. However, the level of disturbance at study sites was a significant predictor of both fuel stores and glycerol levels, suggesting that Dunlins stopping over during spring migration may have had difficulty reaching necessary fuel stores in spring 2011 due to disturbance from cleanup activity on oiled beaches. These effects from disturbance were only observed at sites with high cleanup activity, suggesting that the impact of oil‐spill cleanup on shorebirds may be minimized by limiting cleanup activities to specific areas and times of day.     

Decline in condition of gorgonian octocorals on mesophotic reefs in the northern Gulf of Mexico: before and after the Deepwater Horizon oil spill

Hard-bottom ‘mesophotic’ reefs along the ‘40-fathom’ (73 m) shelf edge in the northern Gulf of Mexico were investigated for potential effects of the Deepwater Horizon (DWH) oil spill from the Macondo well in April 2010. Alabama Alps Reef, Roughtongue Reef, and Yellowtail Reef were near the well, situated 60–88 m below floating oil discharged during the DWH spill for several weeks and subject to dispersant applications. In contrast, Coral Trees Reef and Madison Swanson South Reef were far from the DWH spill site and below the slick for less than a week or not at all, respectively. The reefs were surveyed by ROV in 2010, 2011, and 2014 and compared to similar surveys conducted one and two decades earlier. Large gorgonian octocorals were present at all sites in moderate abundance including Swiftia exserta, Hypnogorgia pendula, Thesea spp., and Placogorgia spp. The gorgonians were assessed for health and condition in a before-after-control-impact (BACI) research design using still images captured from ROV video transects. Injury was modeled as a categorical response to proximity and time using logistic regression. Condition of gorgonians at sites near Macondo well declined significantly post-spill. Before the spill, injury was observed for 4–9 % of large gorgonians. After the spill, injury was observed in 38–50 % of large gorgonians. Odds of injury for sites near Macondo were 10.8 times higher post-spill, but unchanged at far sites. The majority of marked injured colonies in 2011 declined further in condition by 2014. Marked healthy colonies generally remained healthy. Background stresses to corals, including fishing activity, fishing debris, and coral predation, were noted during surveys, but do not appear to account for the decline in condition at study sites near Macondo well.     

Abundance and Size of Gulf Shrimp in Louisiana's Coastal Estuaries following the Deepwater Horizon Oil Spill

The Deepwater Horizon oil spill impacted Louisiana''s coastal estuaries physically, chemically, and biologically. To better understand the ecological consequences of this oil spill on Louisiana estuaries, we compared the abundance and size of two Gulf shrimp species (Farfantepeneus aztecus and Litopeneus setiferus) in heavily affected and relatively unaffected estuaries, before and after the oil spill. Two datasets were used to conduct this study: data on shrimp abundance and size before the spill were available from Louisiana Department of Wildlife and Fisheries (LDWF). Data on shrimp abundance and size from after the spill were independently collected by the authors and by LDWF. Using a Before-After-Control-Impact with Paired sampling (BACIP) design with monthly samples of two selected basins, we found brown shrimp to become more abundant and the mean size of white shrimp to become smaller. Using a BACIP with data on successive shrimp year-classes of multiple basins, we found both species to become more abundant in basins that were affected by the spill, while mean shrimp size either not change after the spill, or increased in both affected and unaffected basins. We conclude that following the oil spill abundances of both species increased within affected estuaries, whereas mean size may have been unaffected. We propose two factors that may have caused these results: 1) exposure to polycyclic aromatic hydrocarbons (PAHs) may have reduced the growth rate of shrimp, resulting in a delayed movement of shrimp to offshore habitats, and an increase of within-estuary shrimp abundance, and 2) fishing closures established immediately after the spill, may have resulted in decreased fishing effort and an increase in shrimp abundance. This study accentuates the complexities in determining ecological effects of oil spills, and the need of studies on the organismal level to reveal cause-and-effect relationships of such events.     

Adrenal Gland and Lung Lesions in Gulf of Mexico Common Bottlenose Dolphins (Tursiops truncatus) Found Dead following the Deepwater Horizon Oil Spill

A northern Gulf of Mexico (GoM) cetacean unusual mortality event (UME) involving primarily bottlenose dolphins (Tursiops truncatus) in Louisiana, Mississippi, and Alabama began in February 2010 and continued into 2014. Overlapping in time and space with this UME was the Deepwater Horizon (DWH) oil spill, which was proposed as a contributing cause of adrenal disease, lung disease, and poor health in live dolphins examined during 2011 in Barataria Bay, Louisiana. To assess potential contributing factors and causes of deaths for stranded UME dolphins from June 2010 through December 2012, lung and adrenal gland tissues were histologically evaluated from 46 fresh dead non-perinatal carcasses that stranded in Louisiana (including 22 from Barataria Bay), Mississippi, and Alabama. UME dolphins were tested for evidence of biotoxicosis, morbillivirus infection, and brucellosis. Results were compared to up to 106 fresh dead stranded dolphins from outside the UME area or prior to the DWH spill. UME dolphins were more likely to have primary bacterial pneumonia (22% compared to 2% in non-UME dolphins, P = .003) and thin adrenal cortices (33% compared to 7% in non-UME dolphins, P = .003). In 70% of UME dolphins with primary bacterial pneumonia, the condition either caused or contributed significantly to death. Brucellosis and morbillivirus infections were detected in 7% and 11% of UME dolphins, respectively, and biotoxin levels were low or below the detection limit, indicating that these were not primary causes of the current UME. The rare, life-threatening, and chronic adrenal gland and lung diseases identified in stranded UME dolphins are consistent with exposure to petroleum compounds as seen in other mammals. Exposure of dolphins to elevated petroleum compounds present in coastal GoM waters during and after the DWH oil spill is proposed as a cause of adrenal and lung disease and as a contributor to increased dolphin deaths.     

Extent and Degree of Shoreline Oiling: Deepwater Horizon Oil Spill,Gulf of Mexico,USA

The oil from the 2010 Deepwater Horizon spill in the Gulf of Mexico was documented by shoreline assessment teams as stranding on 1,773 km of shoreline. Beaches comprised 50.8%, marshes 44.9%, and other shoreline types 4.3% of the oiled shoreline. Shoreline cleanup activities were authorized on 660 km, or 73.3% of oiled beaches and up to 71 km, or 8.9% of oiled marshes and associated habitats. One year after the spill began, oil remained on 847 km; two years later, oil remained on 687 km, though at much lesser degrees of oiling. For example, shorelines characterized as heavily oiled went from a maximum of 360 km, to 22.4 km one year later, and to 6.4 km two years later. Shoreline cleanup has been conducted to meet habitat-specific cleanup endpoints and will continue until all oiled shoreline segments meet endpoints. The entire shoreline cleanup program has been managed under the Shoreline Cleanup Assessment Technique (SCAT) Program, which is a systematic, objective, and inclusive process to collect data on shoreline oiling conditions and support decision making on appropriate cleanup methods and endpoints. It was a particularly valuable and effective process during such a complex spill.     

Flyingfish Spawning (Parexocoetus brachypterus) in the Northeastern Gulf of Mexico

A spawning aggregation of Parexocoetus brachypterus was observed in the northeastern Gulf of Mexico over the outer continental shelf, 100km south of Mobile, Alabama. The event was considered to be a spawning aggregation given the remarkably high abundance of flyingfish observed, together with unusually vigorous flying, jumping, and swimming activity near the surface, and observations of both males and females exuding ripe sex products when landed. The spawn occurred during May 2001 just after moonrise 2 days post full moon. More than 1000000 flyingfish were estimated to have participated in the spawn. Male flyingfish were three times more abundant than females, consistent with observations of 3–4 flyingfish grouped together before and after the spawning aggregation, if several males were simultaneously pursuing a single female. Although egg attachment to flotsam is the only reproductive mode previously described for flyingfishes with demersal eggs, the P. brachypterus spawn occurred in the absence of flotsam. Other possible egg development modes include egg suspension upon current, mid-water suspension above pycnocline or upon topographically induced turbulence, or benthic.     

Interannual patterns of the large free-living nematode assemblages in the Mexican Exclusive Economic Zone,NW Gulf of Mexico after the Deepwater Horizon oil spill

The presence and abundance of particular nematode genera in marine sediments are bioindicators of environmental disturbance. They can even reveal the existence of specific toxicants, such as trace-metals and hydrocarbons derived from fossil fuels. We studied this important infaunal component during a three-year monitoring program in Mexico’s Exclusive Economic Zone (EEZ) in the aftermath of a major oil spill in the northern Gulf of Mexico caused by the sinking of the Deepwater Horizon platform (DWH) in April of 2010. A closer look at the nematode composition and density values throughout and after the DWH oil spill revealed considerable changes. Continental shelf and upper slope sediments sampled in the summer of 2010 included 48 genera and a density of 44.45 ind/10 cm⁻². Eight months later in the winter of 2011, there was a critical decrement in genera number (23) and density (25.22 ind/10 cm⁻²). However, in 2012, the nematode community showed signs of recovery: genus diversity was 58 with a density of 91.45 ind/10 cm⁻². Significant differences were recorded between the first and the last sampling periods, driven by a remarkable turn-over in genera composition, and by an increase in density of the genera Sabatieria, Dorylaimopsis, and Cheironchus. Similarly, significant spatial differences between the nematofauna of the inner-middle shelf and the deepest zone (>2000 m) were detected. Correlation analyses confirmed the tolerant behavior of the above genera to Ni and Co, and their proliferation when hydrocarbon compounds presumably derived from the DWH oil spill increased throughout the study. The Maturity Index (MI) estimated for each survey ranged from 2.35 to 2.6. Environmental Quality Status conditions in the summer of 2010 were good while in the winter of 2011 were poor, and moderate towards the winter of 2012. Index of Trophic Diversity (ITD) values for each survey were high: 0.70, 0.68, and 0.74, respectively. A high predominance of feeding guilds 1B and 2B was recorded throughout the study.The estimated Index of Trophic Diversity (ITD) values resulted high (Table 5). These values apparently indicated that the four feeding types were proportionally distributed in the nematode assemblage. This fact was better reflected during the winter of 2012. Nonetheless, these results are not congruent with the high predominance of feeding guilds 1B and 2B observed throughout the study.     

Detection of Salt Marsh Vegetation Stress and Recovery after the Deepwater Horizon Oil Spill in Barataria Bay,Gulf of Mexico Using AVIRIS Data

The British Petroleum Deepwater Horizon Oil Spill in the Gulf of Mexico was the biggest oil spill in US history. To assess the impact of the oil spill on the saltmarsh plant community, we examined Advanced Visible Infrared Imaging Spectrometer (AVIRIS) data flown over Barataria Bay, Louisiana in September 2010 and August 2011. Oil contamination was mapped using oil absorption features in pixel spectra and used to examine impact of oil along the oiled shorelines. Results showed that vegetation stress was restricted to the tidal zone extending 14 m inland from the shoreline in September 2010. Four indexes of plant stress and three indexes of canopy water content all consistently showed that stress was highest in pixels next to the shoreline and decreased with increasing distance from the shoreline. Index values along the oiled shoreline were significantly lower than those along the oil-free shoreline. Regression of index values with respect to distance from oil showed that in 2011, index values were no longer correlated with proximity to oil suggesting that the marsh was on its way to recovery. Change detection between the two dates showed that areas denuded of vegetation after the oil impact experienced varying degrees of re-vegetation in the following year. This recovery was poorest in the first three pixels adjacent to the shoreline. This study illustrates the usefulness of high spatial resolution airborne imaging spectroscopy to map actual locations where oil from the spill reached the shore and then to assess its impacts on the plant community. We demonstrate that post-oiling trends in terms of plant health and mortality could be detected and monitored, including recovery of these saltmarsh meadows one year after the oil spill.     

Spatial,Temporal, and Habitat-Related Variation in Abundance of Pelagic Fishes in the Gulf of Mexico: Potential Implications of the Deepwater Horizon Oil Spill

Time-series data collected over a four-year period were used to characterize patterns of abundance for pelagic fishes in the northern Gulf of Mexico (GoM) before (2007–2009) and after (2010) the Deepwater Horizon oil spill. Four numerically dominant pelagic species (blackfin tuna, blue marlin, dolphinfish, and sailfish) were included in our assessment, and larval density of each species was lower in 2010 than any of the three years prior to the oil spill, although larval abundance in 2010 was often statistically similar to other years surveyed. To assess potential overlap between suitable habitat of pelagic fish larvae and surface oil, generalized additive models (GAMs) were developed to evaluate the influence of ocean conditions on the abundance of larvae from 2007–2009. Explanatory variables from GAMs were then linked to environmental data from 2010 to predict the probability of occurrence for each species. The spatial extent of surface oil overlapped with early life habitat of each species, possibly indicating that the availability of high quality habitat was affected by the DH oil spill. Shifts in the distribution of spawning adults is another factor known to influence the abundance of larvae, and the spatial occurrence of a model pelagic predator (blue marlin) was characterized over the same four-year period using electronic tags. The spatial extent of oil coincided with areas used by adult blue marlin from 2007–2009, and the occurrence of blue marlin in areas impacted by the DH oil spill was lower in 2010 relative to pre-spill years.     

High Numbers of Vibrio vulnificus in Tar Balls Collected from Oiled Areas of the North-Central Gulf of Mexico Following the 2010 BP Deepwater Horizon Oil Spill

The Deepwater Horizon Oil Spill was the largest oil spill in USA history releasing approximately 4.9 million barrels of crude oil into the Gulf of Mexico. Soon after the spill started, tar balls and other forms of weathered oil appeared in large numbers on beaches in Mississippi and Alabama. In this study, we analyzed tar balls for total aerobic bacterial (TAB) counts and also for the presence of Vibrio vulnificus, a human pathogen known to be abundant in the Gulf Coast environment and capable of causing severe wound infections by contact with contaminated surfaces. Our results showed that TAB counts were significantly higher in tar balls than in sand and seawater collected at the same location. In addition, V. vulnificus numbers were 10× higher in tar balls than in sand and up to 100× higher than in seawater. Densities of V. vulnificus were higher than 10(5) colony forming units/g of tar ball in all samples analyzed. Our data suggest that tar balls can act as reservoirs for bacteria including human pathogens.     

Dramatic shifts in benthic microbial eukaryote communities following the Deepwater Horizon oil spill

Benthic habitats harbour a significant (yet unexplored) diversity of microscopic eukaryote taxa, including metazoan phyla, protists, algae and fungi. These groups are thought to underpin ecosystem functioning across diverse marine environments. Coastal marine habitats in the Gulf of Mexico experienced visible, heavy impacts following the Deepwater Horizon oil spill in 2010, yet our scant knowledge of prior eukaryotic biodiversity has precluded a thorough assessment of this disturbance. Using a marker gene and morphological approach, we present an intensive evaluation of microbial eukaryote communities prior to and following oiling around heavily impacted shorelines. Our results show significant changes in community structure, with pre-spill assemblages of diverse Metazoa giving way to dominant fungal communities in post-spill sediments. Post-spill fungal taxa exhibit low richness and are characterized by an abundance of known hydrocarbon-degrading genera, compared to prior communities that contained smaller and more diverse fungal assemblages. Comparative taxonomic data from nematodes further suggests drastic impacts; while pre-spill samples exhibit high richness and evenness of genera, post-spill communities contain mainly predatory and scavenger taxa alongside an abundance of juveniles. Based on this community analysis, our data suggest considerable (hidden) initial impacts across Gulf beaches may be ongoing, despite the disappearance of visible surface oil in the region.     

The Influence of Habitat Complexity on Reef Fish Communities in the Northeastern Gulf of Mexico

Several previous studies have attempted to correlate habitat complexity and reef fish species diversity. These studies have mostly examined natural reef systems, but results differed. To examine this relation, we built 1 m² habitats with 20 replicates of five complexity levels from July to August 2001 in the northeastern Gulf of Mexico (n=100). In June and July 2002, we built new habitats using the 2001 design, but also added a sixth complexity level (n=120). In order of increasing complexity these included: cage, shell, cage-shell, block-shell, cage-block-shell, and shell-block-pyramid habitats. Most fish in both years were juveniles and included species common to reef structures in the northeastern Gulf of Mexico. In 2001, we identified 26 fish species, and the dominant species was red snapper, Lutjanus campechanus (41%), followed by rock sea bass, Centropristis philadelphica (23%), and sand perch, Diplectrum spp. (14%). In 2002 we identified 36 species, and the dominant species was tomtate, Haemulon aurolineatum (36%), followed by Diplectrum spp. (19%), and L. campechanus (13%). In 2001, species diversity and richness were significantly (P<0.05) higher on more complex habitats (H′=1.7, S=11–12) compared to less complex habitats (H′=0.8–1.0, S=4–9). In 2002, patterns among diversity, richness and reef complexity were less apparent with only the least complex habitats shell and cage showing significantly lower values. In both years, multidimensional scaling grouped by complexity levels with cage and shell habitats showing the clearest separation from other habitat types. Also, with few exceptions (only 8%) analysis of similarities showed significant (P<0.05) differences in fish communities across complexity levels. Although community composition varied between years, this study provided evidence to support the hypothesis that habitat complexity increased reef fish species diversity.     

A suite of citrate-derived siderophores from a marine Vibrio species isolated following the Deepwater Horizon oil spill

Nearly all microbes require iron for growth. The low concentration of iron found in the ocean makes iron acquisition a particularly difficult task. In response to these low iron conditions, many bacteria produce low-molecular-weight iron-binding molecules called siderophores to aid in iron uptake. We report herein the isolation and structural characterization of a suite of amphiphilic siderophores called the ochrobactins-OH, which are produced by a Vibrio species isolated from the Gulf of Mexico after the 2010 Deepwater Horizon oil spill. The citrate-based ochrobactins-OH are derivatives of aerobactin, replacing the acetyl groups with fatty acid appendages ranging in size from C8 to C12, and are distinctly different from the ochrobactins in that the fatty acid appendages are hydroxylated rather than unsaturated. The discovery of the marine amphiphilic ochrobactin-OH suite of siderophores increases the geographic and phylogenetic diversity of siderophore-producing bacteria.     

Regime Shift in Sandy Beach Microbial Communities following Deepwater Horizon Oil Spill Remediation Efforts

Sandy beaches support a wide variety of underappreciated biodiversity that is critical to coastal ecosystems. Prior to the 2010 Deepwater Horizon oil spill, the diversity and function of supratidal beach sediment microbial communities along Gulf of Mexico coastlines were not well understood. As such, it was unclear if microbial community compositional changes would occur following exposure to beached oil, if indigenous communities could biodegrade oil, or how cleanup efforts, such as sand washing and sediment redistribution, would impact microbial ecosystem resiliency. Transects perpendicular to the shoreline were sampled from public beaches on Grand Isle, Louisiana, and Dauphin Island, Alabama, over one year. Prior to oil coming onshore, elevated levels of bacteria associated with fecal contamination were detected (e.g., Enterobacteriales and Campylobacterales). Over time, significant shifts within major phyla were identified (e.g., Proteobacteria, Firmicutes, Actinobacteria) and fecal indicator groups were replaced by taxa affiliated with open-ocean and marine systems (e.g., Oceanospirillales, Rhodospirillales, and Rhodobacterales). These new bacterial groups included putative hydrocarbon degraders, similar to those identified near the oil plume offshore. Shifts in the microbial community composition strongly correlated to more poorly sorted sediment and grain size distributional changes. Natural oceanographic processes could not account for the disrupted sediment, especially from the backshore well above the maximum high-tide levels recorded at these sites. Sand washing and tilling occurred on both open beaches from August through at least December 2010, which were mechanisms that could replace fecal indicator groups with open-ocean groups. Consequently, remediation efforts meant to return beaches to pre-spill compositions caused a regime shift that may have added potential ecosystem function, like hydrocarbon degradation, to the sediment. Future research will need to assess the persistence and impact of the newly formed microbial communities to the overall sandy beach ecosystems.     

Disturbance and recovery of salt marsh arthropod communities following BP Deepwater Horizon oil spill

Oil spills represent a major environmental threat to coastal wetlands, which provide a variety of critical ecosystem services to humanity. The U.S. Gulf of Mexico is a hub of oil and gas exploration activities that historically have impacted intertidal habitats such as salt marsh. Following the BP Deepwater Horizon oil spill, we sampled the terrestrial arthropod community and marine invertebrates found in stands of Spartina alterniflora, the most abundant plant in coastal salt marshes. Sampling occurred in 2010 as oil was washing ashore and a year later in 2011. In 2010, intertidal crabs and terrestrial arthropods (insects and spiders) were suppressed by oil exposure even in seemingly unaffected stands of plants; however, Littoraria snails were unaffected. One year later, crab and arthropods had largely recovered. Our work is the first attempt that we know of assessing vulnerability of the salt marsh arthropod community to oil exposure, and it suggests that arthropods are both quite vulnerable to oil exposure and quite resilient, able to recover from exposure within a year if host plants remain healthy.     

Culture-Independent Characterization of Bacterial Communities Associated with the Cold-Water Coral Lophelia pertusa in the Northeastern Gulf of Mexico

Bacteria are recognized as an important part of the total biology of shallow-water corals. Studies of shallow-water corals suggest that associated bacteria may benefit the corals by cycling carbon, fixing nitrogen, chelating iron, and producing antibiotics that protect the coral from other microbes. Cold-water or deep-sea corals have a fundamentally different ecology due to their adaptation to cold, dark, high-pressure environments and as such have novel microbiota. The goal of this study was to characterize the microbial associates of Lophelia pertusa in the northeastern Gulf of Mexico. This is the first study to collect the coral samples in individual insulated containers and to preserve coral samples at depth in an effort to minimize thermal shock and evaluate the effects of environmental gradients on the microbial diversity of samples. Molecular analysis of bacterial diversity showed a marked difference between the two study sites, Viosca Knoll 906/862 (VK906/862) and Viosca Knoll 826 (VK826). The bacterial communities from VK826 were dominated by a variety of unknown mycoplasmal members of the Tenericutes and Bacteroidetes, whereas the libraries from VK906/862 were dominated by members of the Proteobacteria. In addition to novel sequences, the 16S rRNA gene clone libraries revealed many bacterial sequences in common between Gulf of Mexico Lophelia corals and Norwegian fjord Lophelia corals, as well as shallow-water corals. Two Lophelia-specific bacterial groups were identified: a cluster of gammaproteobacteria related to sulfide-oxidizing gill symbionts of seep clams and a group of Mycoplasma spp. The presence of these groups in both Gulf and Norwegian Lophelia corals indicates that in spite of the geographic heterogeneity observed in Lophelia-associated bacterial communities, there are Lophelia-specific microbes.Cold-water and deep-sea corals have become a topic of interest due to conservation concerns over the impacts of trawling, exploration for oil and gas, and climate change (51, 52). Although the existence of these corals has been known since the 1800s, our knowledge of their distribution, ecology, and biology is limited due to the technical difficulties of studying them. Lophelia pertusa is a globally distributed cold-water scleractinian coral (53). In the Gulf of Mexico, Lophelia reefs occur primarily along the continental shelf break (300- to 500-m depth), providing an important complex habitat for a wide variety of fishes, crustaceans, and other invertebrates living below the photic zone (48).The microbial ecology of cold-water corals in deep water is fundamentally different from that of shallow-water corals due to the ambient environmental parameters (e.g., darkness, low temperature, and increased pressure) and the absence of symbiotic zooxanthellae. A few studies have begun to address the microbial associates of deep-sea corals, focusing on octocorals (9, 44) and on L. pertusa (27, 41, 42, 57, 72). To date, all the Lophelia studies have been conducted on the eastern side of the Atlantic: the Mediterranean basin (72), Mingulay Bay, Scotland (27), and Norwegian fjords (41, 42, 57). These studies have confirmed that the Lophelia-associated bacterial community is distinct from that of the surrounding seawater and sediments (27, 42, 57, 72). A variety of community profile methods (automated rRNA intergenic spacer analysis, terminal restriction fragment length polymorphism, and denaturing gradient gel electrophoresis [DGGE]) were used to demonstrate differences between samples within a geographic area, suggesting that the Lophelia-associated microbial community varies depending on regional environmental factors (27, 42, 57). Sequencing of 16S rRNA genes was done in only two studies, and there was no overlap between their data (42, 72). However, different methods of collection, extraction, amplification, and sequencing were employed, so the lack of commonality may be due to methodology rather than biogeography.Methodology is a concern, particularly the care with which samples need to be collected for microbial ecology studies. Deep-sea coral samples are typically collected by a trawl, net, or dredge or by a submersible/remotely operated vehicle (ROV). With these methods, many corals may be combined in a single container, which is not acceptable for microbiological studies because the microbial community of one coral could contaminate that of the other. Similarly, contact with sediment, other invertebrates, mobile fauna, or water masses between the collection point and the surface could contaminate the coral samples. Unlike the case with the northeastern Atlantic and Norwegian fjords, the temperature and salinity gradients in the Gulf of Mexico during the warm months of the year can be considerable. In the case of the Viosca Knoll sites, the bottom temperature was 8 to 11°C, compared to a surface temperature of ≥30°C. Coral samples collected in uninsulated containers in this area have been observed to be affected (e.g., polyps retracted and copious stress mucus production) compared to those in insulated containers. Viosca Knoll is also impacted by the Mississippi River plume. The surface waters at these sites were turbid and green and had a salinity of 30 practical salinity units (psu), but below the plume the waters were clear and had a salinity of 35 psu. With this in mind, we designed a sampling container that would protect the coral samples from dramatic changes in temperature and salinity by sealing them in individual insulated compartments (see Fig. S1 in the supplemental material). However, the question remained whether environmental gradients in light and pressure would have an effect on the microbial diversity of the samples. To address this question, each sample was collected in duplicate: one piece was sealed in a compartment alive, and a replicate piece was sealed in another compartment and preserved at depth with a fixative solution. Both sample types (“live” versus “fixed”) were sealed and insulated, so temperature and salinity gradients did not affect them; live samples were subject to gradients in light and pressure, while fixed samples were not.The main objective of this study was to characterize the bacterial associates of Lophelia pertusa from two sites in the northern Gulf of Mexico. Comparing multiple individual colonies from two geographic locations in the Gulf to each other and to bacterial data from Lophelia samples on the eastern side of the Atlantic will clarify whether Lophelia has a species-specific bacterial community, as has been described for shallow-water corals (49, 55). The results of this study will also better define the total microbial diversity associated with this cold-water coral. A specialized sampling device (see Fig. S1 in the supplemental material) was designed to minimize contamination and thermal shock and to allow the introduction of preservative at depth to determine if environmental gradients were affecting microbial diversity during sampling.