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Wastewater contamination threatens the shellfish aquaculture industry by posing risks to public health. Multiple indicators of wastewater contamination, including fecal coliforms (fc), male-specific coliphage (MSC), dissolved nutrients, stable isotope ratios, and artificial sweeteners were analyzed to determine possible sources of wastewater to local shellfish farms. Samples were collected at a wastewater treatment plant outfall (WTPO), nonpoint residential, and agricultural areas of a tidal river, and tidal creek inflows adjacent to farms. To capture seasonal variation, we sampled under warm and cold, and wet and dry conditions. Fc ranged −1, NH4+ concentrations ranged up to 9.58 μM, and δ15N ranged 1.4–7.8‰ across all sites and time periods. Fc and NH4+ were higher, and δ15N was lower in the cold wet period and near residential and agricultural areas. Acesulfame and sucralose concentrations ranged 0.004–0.05 μg L−1 and up to > 0.8 μg L−1, respectively, and did not correlate with other indicators but tended to be higher in residential areas and at the WTPO, supporting their value in differentiating human sewage from other sources. Shoreline disturbance during septic system upgrades may have inadvertently contributed bacterial indicators to shellfish farms. Overall, indicator source dominance depended on environmental conditions, with WTPO and residential sources conveying human-specific indicators to farms year-round, while agricultural and industrial sites contributed additional fc during cold wet periods. The use of multiple indicators will aid managers to detect and define wastewater sources, identify targets for monitoring or remediation, and manage shellfish areas in estuaries with a mosaic of land-derived wastewater sources.

Freshwater-dominated estuaries experience large fluctuations in their physical and chemical environments which may influence larval dispersal, settlement, and connectivity of populations with pelagic larval stages. We used settlement patterns and natural tagging along with numerical hydrodynamic model results to assess settlement and connectivity among oysters across the freshwater-dominated Mobile Bay-eastern Mississippi Sound (MB-EMS) system. Specifically, we (1) tested how freshwater inputs and associated environmental attributes influenced settlement patterns during high and low discharge conditions in 2014 and 2016, respectively, and (2) analyzed trace element (TE) ratios incorporated into multiple shell types (larval and settled shell of spat and adult shells) to determine if shells collected in situ incorporate temporally stable site-specific signatures. We also assessed if TE ratios compared between adult (TE natal signature proxy) and larval shells could infer connectivity. Larval settlement was 4× higher during low discharge than during high discharge when oyster larvae only settled in higher salinity regions (EMS). Spat and adult shells incorporated site-specific TE ratios that varied from weeks to months. Connectivity results (May-June 2016 only) suggest that EMS is an important larval source to EMS and lower MB. While we were able to infer probable connectivity patterns using adult and larval shells, more study is needed to assess the utility of adult shells as proxies for natal-location TE signatures. Results provide a baseline for measuring future larval connectivity and adult distribution changes in the MB-EMS system. Biological and geochemical data demonstrate the potential to identify environmental attributes that spatiotemporally mediate settlement and connectivity in dynamic systems.

Dolphin morbillivirus (DMV) is a virulent pathogen that causes high mortality outbreaks in delphinids globally and is spread via contact among individuals. Broadly ranging nearshore and open-ocean delphinids are likely reservoir populations that transmit DMV to estuarine populations. We assessed the seroprevalence of DMV antibodies and determined the habitat use of common bottlenose dolphins, Tursiops truncatus truncatus, from two estuarine sites, Barataria Bay and Mississippi Sound, in the northern Gulf of Mexico. We predicted that risk to DMV exposure in estuarine dolphins is driven by spatial overlap in habitat use with reservoir populations. Serum was collected from live-captured dolphins and tested for DMV antibodies. Habitat use of sampled individuals was determined by analysing satellite-tracked movements and stable isotope values. DMV seroprevalences were high among dolphins at Barataria Bay (37%) and Mississippi Sound (44%), but varied differently within sites. Ranging patterns of Barataria Bay dolphins were categorized into two groups: Interior and Island-associated. DMV seroprevalences were absent in Interior dolphins (0%) but high in Island-associated dolphins (45%). Ranging patterns of Mississippi Sound dolphins were categorized into three groups: Interior, Island-east and Island-west. DMV seroprevalences were detected across Mississippi Sound (Interior: 60%; Island-east: 20%; and Island-west: 43%). At both sites, dolphins in habitats with greater marine influence had enriched δ13 C values, and Barataria Bay dolphins with positive DMV titres had carbon isotope values indicative of marine habitats. Positive titres for DMV antibodies were more common in the lower versus upper parts of Barataria Bay but evenly distributed across Mississippi Sound. A dolphin's risk of exposure to DMV is influenced by how individual ranging patterns interact with environmental geography. Barataria Bay's partially enclosed geography likely limits the nearshore or open-ocean delphinids that carry DMV from interacting with dolphins that use interior, estuarine habitats, decreasing their exposure to DMV. Mississippi Sound's relatively open geography allows for greater spatial overlap and mixing among estuarine, nearshore and/or open-ocean cetaceans. The spread of DMV, and likely other diseases, is affected by the combination of individual movements, habitat use and the environment.

Recognizing the imperative to evaluate species recovery and conservation impact, in 2012 the International Union for Conservation of Nature (IUCN) called for development of a "Green List of Species" (now the IUCN Green Status of Species). A draft Green Status framework for assessing species' progress toward recovery, published in 2018, proposed 2 separate but interlinked components: a standardized method (i.e., measurement against benchmarks of species' viability, functionality, and preimpact distribution) to determine current species recovery status (herein species recovery score) and application of that method to estimate past and potential future impacts of conservation based on 4 metrics (conservation legacy, conservation dependence, conservation gain, and recovery potential). We tested the framework with 181 species representing diverse taxa, life histories, biomes, and IUCN Red List categories (extinction risk). Based on the observed distribution of species' recovery scores, we propose the following species recovery categories: fully recovered, slightly depleted, moderately depleted, largely depleted, critically depleted, extinct in the wild, and indeterminate. Fifty-nine percent of tested species were considered largely or critically depleted. Although there was a negative relationship between extinction risk and species recovery score, variation was considerable. Some species in lower risk categories were assessed as farther from recovery than those at higher risk. This emphasizes that species recovery is conceptually different from extinction risk and reinforces the utility of the IUCN Green Status of Species to more fully understand species conservation status. Although extinction risk did not predict conservation legacy, conservation dependence, or conservation gain, it was positively correlated with recovery potential. Only 1.7% of tested species were categorized as zero across all 4 of these conservation impact metrics, indicating that conservation has, or will, play a role in improving or maintaining species status for the vast majority of these species. Based on our results, we devised an updated assessment framework that introduces the option of using a dynamic baseline to assess future impacts of conservation over the short term to avoid misleading results which were generated in a small number of cases, and redefines short term as 10 years to better align with conservation planning. These changes are reflected in the IUCN Green Status of Species Standard.Reconociendo que era imperativo evaluar la recuperación de especies y el impacto de la conservación, la Unión Internacional para la Conservación de la Naturaleza (UICN) convocó en 2012 al desarrollo de una “Lista Verde de Especies” (ahora el Estatus Verde de las Especies de la UICN). Un marco de referencia preliminar de una Lista Verde de Especies para evaluar el progreso de las especies hacia la recuperación, publicado en 2018, proponía 2 componentes separados pero interconectados: un método estandarizado (i.e., medición en relación con puntos de referencia de la viabilidad de especies, funcionalidad y distribución antes del impacto) para determinar el estatus de recuperación actual (puntuación de recuperación de la especie) y la aplicación de ese método para estimar impactos en el pasado y potenciales de conservación basados en 4 medidas (legado de conservación, dependencia de conservación, ganancia de conservación y potencial de recuperación). Probamos el marco de referencia con 181 especies representantes de diversos taxa, historias de vida, biomas, y categorías (riesgo de extinción) en la Lista Roja de la IUCN. Con base en la distribución observada de la puntuación de recuperación de las especies, proponemos las siguientes categorías de recuperación de la especie: totalmente recuperada, ligeramente mermada, moderadamente mermada, mayormente mermada, gravemente mermada, extinta en estado silvestre, e inderterminada. Cincuenta y nueve por ciento de las especies se consideraron mayormente o gravemente mermada. Aunque hubo una relación negativa entre el riesgo de extinción y la puntuación de recuperación de la especie, la variación fue considerable. Algunas especies en las categorías de riesgo bajas fueron evaluadas como más lejos de recuperarse que aquellas con alto riesgo. Esto enfatiza que la recuperación de especies es diferente conceptualmente al riesgo de extinción y refuerza la utilidad del Estado Verde de las Especies de la UICN para comprender integralmente el estatus de conservación de especies. Aunque el riesgo de extinción no predijo el legado de conservación, la dependencia de conservación o la ganancia de conservación, se correlacionó positivamente con la potencial de recuperación. Solo 1.7% de las especies probadas fue categorizado como cero en los 4 indicadores de impacto de la conservación, lo que indica que la conservación ha jugado, o jugará, un papel en la mejoría o mantenimiento del estatus de la especie la gran mayoría de ellas. Con base en nuestros resultados, diseñamos una versión actualizada del marco de referencia para la evaluación que introduce la opción de utilizar una línea de base dinámica para evaluar los impactos futuros de la conservación en el corto plazo y redefine corto plazo como 10 años.

Mobile, apex predators are commonly assumed to stabilize food webs through trophic coupling across spatially distinct habitats. The assumption that trophic coupling is common remains largely untested, despite evidence that individual behaviors might limit trophic coupling. We used stable isotope data from common bottlenose dolphins across the Gulf of Mexico to determine if these apex predators coupled estuarine and adjacent, nearshore marine habitats. δ13C values differed among the sites, likely driven by environmental factors that varied at each site, such as freshwater input and seagrass cover. Within most sites, δ13C values differed such that dolphins sampled in the upper reaches of embayments had values indicative of estuarine habitats while those sampled outside or in lower reaches of embayments had values indicative of marine habitats. δ15N values were more similar among and within sites than δ13C values. Data from multiple tissues within individuals corroborated that most dolphins consistently used a narrow range of habitats but fed at similar trophic levels in estuarine and marine habitats. Because these dolphins exhibited individual habitat specialization, they likely do not contribute to trophic coupling between estuarine and adjacent marine habitats at a regional scale, suggesting that not all mobile, apex predators trophically couple adjacent habitats.

Ecosystems are experiencing elevated levels of disturbance, and species with narrower niches are often more vulnerable to disturbances. Niche breadth is often measured in terms of either diet or habitat use but diet and spatial use are infrequently considered in tandem. These different aspects of niche breadth potentially expose species to different types of disturbances; species with narrow dietary niches may be more affected by disturbances that alter trophic relationships, while species with narrow habitat niches may be more vulnerable to habitat loss and point‐source pollutants. We examined dietary and habitat niche breadth of common bottlenose dolphins, Tursiops truncatus truncatus, from three different nearshore sites in the Gulf of Mexico (GoM). Using stable isotopes, we determined proportional contributions of different prey groups to dolphin diets at each site and through time at one site. We used satellite‐linked telemetry at two sites to determine habitat use and site fidelity. Additionally, we examined the literature on cetacean diet, habitat use, movement, and IUCN status to determine relationships between niche breadth and population status for different species. Dolphin diets varied among sites as available prey varied, but Perciformes fish were the most frequently consumed prey. At the site for which we had temporal data, dolphins consumed more cephalopods in 2015 and 2018 but otherwise consumed primarily Perciformes fish. Dolphins had small utilization distributions and exhibited high site fidelity. Data from 31 cetacean species revealed that most species with vulnerable, threatened, or endangered IUCN statuses not only have specialized diets but also exhibit high site fidelity. Dolphins had diet characteristic of flexible generalists but were habitat specialists with high site fidelity. Dolphin populations in the GoM may have altered their diets in response to environmental changes that have altered community composition and trophic dynamics. On the other hand, their high site fidelity has exposed them to point‐source pollutants, such as oil spills, persistent organic pollutants, and freshwater. Our broader analysis of cetaceans confirmed that species with specialized diets and high site fidelity were the most vulnerable to disturbances, providing a framework to predict which nearshore dolphin populations, and cetaceans in general, are most vulnerable to environmental changes.

Knowledge of larval transport is important for restoration and management efforts; yet, there are no established methods to determine larval transport in situ. Calcein staining of oyster larvae may help fill this void, and a two-part study was conducted to determine its effectiveness at tracking larval oyster transport in the field. First, it was tested whether oysters could be successfully stained, survive, and grow at estuarine salinities (15, 20, 26), and at sufficiently large numbers (millions of oysters) to support field mark-recapture studies. Second, the field-based application was tested by releasing 22 million stained larvae twice (high and low salinity) into a major estuary, and two methods (fluorescent microscopy and FlowCam) were used to detect recaptured larvae. Results were compared with expected larval movement patterns simulated by an existing larval transport model. Calcein concentrations (100 mg L−1) did not affect larval growth or survival, but handling conditions (water salinity manipulations and tank size) did affect growth and survival during the post-staining period. Microscopy had double the detection capacity, but FlowCam was more practical and time efficient for the large-volume, high particulate load field samples. Larvae (n = 2) were recaptured during the second, higher salinity release, and model comparison showed a 1–2-day time-lag between field recapture and model predictions, suggesting need for model refinement. Calcein has potential to be a useful marker to track larval movement at large scales needed for field-based studies, providing critical information to aid in selection of restoration sites and management of commercially important shellfish species.

Collaboration among scientists and stakeholders is increasingly valued in research to coproduce knowledge and research products that better inform decision making and enact meaningful change. We present an example of effective coproduction of knowledge to protect water quality along the Mississippi–Alabama coast using a comprehensive approach that tracked progress from initial research through product assessment. We coproduced an education and decision support tool known as “Our Wastewater Footprint” and engaged communities through a variety of public outreach efforts, adapting the product to meet the needs of individual end users. We assessed the effectiveness of our efforts by tracking attendance at outreach activities, measuring website traffic, and collecting survey data from end users after product use. Data from >9,900 users indicated that presentations at community events and print and social media posts most efficiently reached large audiences using limited resources, and social media posts were most effective in promoting changes in behavior and attitudes on a social level. This case study exemplifies how involving stakeholders in research and product development can increase community engagement in stewardship and prompt change to enhance water quality. Our results tangibly demonstrate that meaningful assessment of the administrative and social impacts of coproduced knowledge is feasible and can be accomplished in a short period of time.

Continental-margin sediments account for ~ 50% of the oceanic biogenic silica burial despite covering

Traditional research methods are often limited in their ability to capture broad spatial and temporal changes in species distribution that affect global patterns of biodiversity. To provide range-wide demographic data needed to quantify and evaluate changes in habitat use and support ongoing recovery efforts for the endangered West Indian manatee Trichechus manatus, we analyzed data from a formal manatee sighting network that uses citizen-sourced data for the understudied north-central Gulf of Mexico region. Although historically considered to be outside manatees’ typical US range, more than 1700 opportunistic, publicly reported manatee sightings and 23 mortalities have been documented in Alabama and Mississippi since the early 1900s. Live manatee sightings have occurred primarily during warmer months in rivers and subembayments. Manatee mortalities have significantly increased since the mid-1980s and have most often been attributed to cold stress, with 2 known mortalities due to vessel strikes in recent years. Sightings of individual manatees were most common, but group sizes of up to 17 were reported, typically in late summer. Decadal-scale trends in opportunistic sighting records demonstrate persistent spatial and temporal patterns of manatee occurrence in the north-central Gulf of Mexico and suggest greater use and importance of the region as seasonal manatee habitat than previously documented. If applied appropriately, citizen-sourced data have the potential to enhance targeted research efforts, significantly contribute to ecological datasets for a number of species, and provide a useful tool to enhance conservation and management.

Oysters (Crassostrea virginica) were an important food resource for native peoples of the northern Gulf of Mexico, who deposited waste shells in middens. Nitrogen (N) stable isotopes (δ15N) in bivalve shells have been used as modern proxies for estuarine N sources because they approximate δ15N in suspended particulate matter. We tested the use of midden shell δ15N as a proxy for ancient estuarine N sources. We hypothesized that isotopic signatures in ancient shells from coastal Mississippi would differ from modern shells due to increased anthropogenic N sources, such as wastewater, through time. We decalcified shells using an acidification technique previously developed for modern bivalves, but modified to determine δ15N, δ13C, %N, and % organic C of these low-N, high-C specimens. The modified method resulted in the greatest percentage of usable data from midden shells. Our results showed that oyster shell δ15N did not significantly differ between ancient (500–2100 years old) and modern oysters from the same locations where the sites had undergone relatively little land-use change. δ15N values in modern shells, however, were positively correlated with water column nitrate concentrations associated with urbanization. When N content and total shell mass were combined, we estimated that middens sequestered 410–39,000 kg of relic N, buried at a rate of up to 5 kg N m−2 yr−1. This study provides a relatively simple technique to assess baseline conditions in ecosystems over long time scales by demonstrating that midden shells can be an indicator of pre-historic N source to estuaries and are a potentially significant but previously uncharacterized estuarine N sink.

The potential role of morbillivirus was evaluated in the deaths of >1100 bottlenose dolphins Tursiops truncatus and other small cetaceans that stranded from February 2010 through July 2014, during the northern Gulf of Mexico (GoM) unusual mortality event (UME). Morbillivirus analysis was carried out on 142 live or freshly dead cetaceans and results were combined with samples from 102 live, free-ranging bottlenose dolphins sampled during capture-release health assessments conducted from 2011 to 2014. Polymerase chain reaction (PCR) testing for morbillivirus showed that 9.9% (14/142) of the stranded cetaceans and 1% (1/83) of the free-ranging live dolphins were positive for dolphin morbilliviral (DMV) RNA. In contrast, previous DMV dolphin die-offs had DMV detectable by PCR in 61 to 97% of animals tested. Histologic findings consistent with morbillivirus infection, including lymphoid depletion, bronchointerstitial pneumonia, syncytial cell formation, or meningoencephalitis, were found in 6.6% (9/136) of the cetaceans that underwent histologic examinations. Serological analysis using a virus neutralization assay found that 29% (5/17) of live stranded and 23% (23/102) of live free-ranging bottlenose dolphins had titers of 64 or greater for cetacean morbillivirus, indicating prior but not necessarily recent exposure to morbillivirus. Current findings suggest that DMV infection, although present in the northern GoM, was sporadic and occurred at low levels and therefore was not the primary cause of the northern GoM UME. Confirmation of DMV infections and existing DMV titers demonstrate continued exposure to morbillivirus among northern GoM cetaceans since the first detection of this virus in the early 1990s.

The potential for stranded dolphins to serve as a tool for monitoring free-ranging populations would be enhanced if their stocks of origin were known. We used stable isotopes of carbon, nitrogen, and sulfur from skin to assign stranded bottlenose dolphins Tursiops truncatus to different habitats, as a proxy for stocks (demographically independent populations), following the Deepwater Horizon oil spill. Model results from biopsy samples collected from dolphins from known habitats (n = 205) resulted in an 80.5% probability of correct assignment. These results were applied to data from stranded dolphins (n = 217), resulting in predicted assignment probabilities of 0.473, 0.172, and 0.355 to Estuarine, Barrier Island (BI), and Coastal stocks, respectively. Differences were found west and east of the Mississippi River, with more Coastal dolphins stranding in western Louisiana and more Estuarine dolphins stranding in Mississippi. Within the Estuarine East Stock, 2 groups were identified, one predominantly associated with Mississippi and Alabama estuaries and another with western Florida. δ15N values were higher in stranded samples for both Estuarine and BI stocks, potentially indicating nutritional stress. High probabilities of correct assignment of the biopsy samples indicate predictable variation in stable isotopes and fidelity to habitat. The power of δ34S to discriminate habitats relative to salinity was essential. Stable isotopes may provide guidance regarding where additional testing is warranted to confirm demographic independence and aid in determining the source habitat of stranded dolphins, thus increasing the value of biological data collected from stranded individuals. Publisher PDF

There have been efforts around the globe to track individuals of many marine species and assess their movements and distribution, with the putative goal of supporting their conservation and management. Determining whether, and how, tracking data have been successfully applied to address real-world conservation issues is, however, difficult. Here, we compile a broad range of case studies from diverse marine taxa to show how tracking data have helped inform conservation policy and management, including reductions in fisheries bycatch and vessel strikes, and the design and administration of marine protected areas and important habitats. Using these examples, we highlight pathways through which the past and future investment in collecting animal tracking data might be better used to achieve tangible conservation benefits.

Telemetry is a key, widely used tool to understand marine megafauna distribution, habitat use, behavior, and physiology; however, a critical question remains: “How many animals should be tracked to acquire meaningful data sets?” This question has wide-ranging implications including considerations of statistical power, animal ethics, logistics, and cost. While power analyses can inform sample sizes needed for statistical significance, they require some initial data inputs that are often unavailable. To inform the planning of telemetry and biologging studies of marine megafauna where few or no data are available or where resources are limited, we reviewed the types of information that have been obtained in previously published studies using different sample sizes. We considered sample sizes from one to >100 individuals and synthesized empirical findings, detailing the information that can be gathered with increasing sample sizes. We complement this review with simulations, using real data, to show the impact of sample size when trying to address various research questions in movement ecology of marine megafauna. We also highlight the value of collaborative, synthetic studies to enhance sample sizes and broaden the range, scale, and scope of questions that can be answered., A. M. M. Sequeira was supported by an ARC Grant (DE170100841), and the Australian Institute of Marine Science, G.C. Hays by the Bertarelli Foundation as part of the Bertarelli Programme in Marine Science, and H. J. Calich by an Australian Government RTP scholarship at UWA. Workshop funding was granted to M. Thums, A. M. M. Sequeira, and C. M. Duarte by the UWA Oceans Institute, the Australian Institute of Marine Science, and the Office of Sponsored Research at King Abdullah University of Science and Technology (KAUST).

This study provides regional-scale data on drivers of horseshoe crab (Limulus polyphemus) presence along the northcentral Gulf of Mexico coast and has implications for understanding habitat suitability for sparse horseshoe crab populations of conservation concern worldwide. To collect baseline data on the relationship between environmental factors and presence of horseshoe crabs, we surveyed four sites from the Fort Morgan peninsula of Mobile Bay, Alabama (AL) to Horn Island, Mississippi (MS). We documented number, size and sex of live animals, molts, and carcasses as metrics of horseshoe crab presence and demographics for two years. Data were compared to in situ and remotely sensed environmental attributes to assess environmental drivers of occurrence during the time of study. Overall, greater evidence of horseshoe crab presence was found at western sites (Petit Bois and Horn Islands) compared to eastern sites (Dauphin Island, Fort Morgan peninsula), mediated by a combination of distance from areas of high freshwater discharge and interannual variation in weather. Higher sex ratios also were found associated with higher occurrence, west of Mobile Bay. Land cover, particularly Bare Land and Estuarine Emergent Wetland classes that are common to western sites, was most predictive of live animal and to some extent carcass occurrence. Our findings suggest that small-scale variation in habitat quality can affect occurrence of horseshoe crabs in sparse populations where density is not a limiting factor. Data from molts and carcasses were informative to supplement live animal data and may be useful to enhance ecological assessment and support conservation and management in regions with sparse populations.

Effects of land use on hydrology, organic matter sources, and processing may be proportionately greater in tidal creeks than large estuaries, yet tidal creek systems have been undervalued in assessments of emerging effects of anthropogenic land use. Through sampling of dated sediment cores, we identified indicators of historical land use change (past 150 yr) in a microtidal northern Gulf of Mexico tidal creek system in the early stages of urbanization. We found that tidal creeks differed from open water sites, and urbanized sites differed from less altered sites, primarily due to changes in carbon sources indicated by differences in sediment stable carbon isotope (δ13C) values, and concentrations of fecal indicator bacterium Clostridium perfringens. Total organic carbon (%TOC) and carbon : nitrogen (C : N) increased twofold in tidal creeks during upstream urbanization in the early 20th century, which led to elevated mid-century sediment TOC accumulation rates (5–10 mg C cm−2 yr−1), followed by decreases in TOC accumulation in tidal creeks and open waters since the 1960s (0.4–1.8 mg C cm−2 yr−1). C. perfringens and nitrogen stable isotope values (δ15N) were, respectively, 1.8 and 1.5 times higher at wastewater-influenced sites than at other sites, increasing through time or remaining high at wastewater-influenced sites from approximately the 1950s-present, when human populations quadrupled. Hence, urbanization altered estuarine inputs from upland C sources and increased inputs from human-derived N and microbes. These findings suggest that tidal creeks are more sensitive archives of land-use change than open water systems due to their proximity and greater connectivity to the watershed.

Rationale Extraction of lipids from tissues prior to carbon stable isotope analysis (SIA) has become a common practice, despite a lack of species-specific data to indicate when lipid extraction is needed. Marine invertebrates, including bivalves, are known to store carbon as glycogen and less in the form of lipids than other species, potentially reducing the need for lipid extraction even when C:N values are above 3.5, a value that previous studies suggest indicates a need for lipid extraction of animal tissues. Methods We investigated the need for lipid extraction on individual tissues (adductor muscle, gut gland, gill) and whole tissue of a glycogen-storing species, the oyster Crassostrea virginica. Bulk and lipid-extracted samples were analyzed for their C and N stable isotope ratios by continuous flow isotope ratio mass spectrometry (IRMS). Samples were analyzed on a 20–20 isotope ratio mass spectrometer (PDZ Europa) after combustion in an elemental analyzer (PDZ Europa Automatic Analyzer-Gas Solid Liquid). Results Although the C:N values for most bulk (unextracted) tissue samples were greater than 3.5, the lipid-extracted δ13C values did not differ from the bulk values. Lipid extraction, however, affected δ15N values in all tissue types except adductor muscle, indicating that separate SIA may be required when tissues are lipid extracted. Conclusions These data demonstrate that it is not necessary to lipid extract oyster tissues in all cases, and that C:N thresholds for lipid extraction in other species may not be reliable for organisms such as oysters that store glycogen. Our data indicate that minimizing unnecessary lipid extraction through preliminary testing will save researchers time and expense by avoiding superfluous sample handling, reducing concern over secondary effects on data quality, and reducing the costs of reagents and additional separate stable isotope analysis to ensure analytical accuracy. Copyright © 2016 John Wiley & Sons, Ltd.

The extent of increasing anthropogenic impacts on large marine vertebrates partly depends on the animals’ movement patterns. Effective conservation requires identification of the key drivers of movement including intrinsic properties and extrinsic constraints associated with the dynamic nature of the environments the animals inhabit. However, the relative importance of intrinsic versus extrinsic factors remains elusive. We analyze a global dataset of ∼2.8 million locations from >2,600 tracked individuals across 50 marine vertebrates evolutionarily separated by millions of years and using different locomotion modes (fly, swim, walk/paddle). Strikingly, movement patterns show a remarkable convergence, being strongly conserved across species and independent of body length and mass, despite these traits ranging over 10 orders of magnitude among the species studied. This represents a fundamental difference between marine and terrestrial vertebrates not previously identified, likely linked to the reduced costs of locomotion in water. Movement patterns were primarily explained by the interaction between species-specific traits and the habitat(s) they move through, resulting in complex movement patterns when moving close to coasts compared with more predictable patterns when moving in open oceans. This distinct difference may be associated with greater complexity within coastal microhabitats, highlighting a critical role of preferred habitat in shaping marine vertebrate global movements. Efforts to develop understanding of the characteristics of vertebrate movement should consider the habitat(s) through which they move to identify how movement patterns will alter with forecasted severe ocean changes, such as reduced Arctic sea ice cover, sea level rise, and declining oxygen content., Workshop funding was granted by the University of Western Australia (UWA) Oceans Institute, the Australian Institute of Marine Science (AIMS), and King Abdullah University of Science and Technology (KAUST). A.M.M.S. was supported by Australian Research Council Grant DE170100841 and an Indian Ocean Ocean Marine Research Centre (UWA, AIMS, Commonwealth of Scientific and Industrial Research Organisation) fellowship. J.P.R., V.M.E., and J.F.G. were supported by Agencia Estatal de Investigación (AEI, Spain) and Fondo Europeo de Desarrollo Regional (FEDER) through project Spatiotemporality in Sociobological Interactions, Models and Methods (SPASIMM) (FIS2016-80067-P AEI/FEDER, European Union), and by research funding from KAUST. J.P.R. was supported by Ministerio de Educación, Cultura y Deporte (Formación de Profesorado Universitario Grant, Spain). D.W.S. was supported by the UK Natural Environment Research Council and Save Our Seas Foundation. N.Q. was supported by Fundação para a Ciência e Tecnologia (Portugal). M.M.C.M. was supported by a Coordenação de Aperfeiçoamento de pessoal de Nível Superior fellowship (Ministry of Education).

An unusual mortality event (UME) was declared for cetaceans in the northern Gulf of Mexico (GoM) for Franklin County, Florida, west through Louisiana, USA, beginning in February 2010 and was ongoing as of September 2014. The 'Deepwater Horizon' (DWH) oil spill began on 20 April 2010 in the GoM, raising questions regarding the potential role of the oil spill in the UME. The present study reviews cetacean mortality events that occurred in the GoM prior to 2010 (n = 11), including causes, durations, and some specific test results, to provide a historical context for the current event. The average duration of GoM cetacean UMEs prior to 2010 was 6 mo, and the longest was 17 mo (2005-2006). The highest number of cetacean mortalities recorded during a previous GoM event was 344 (in 1990). In most previous events, dolphin morbillivirus or brevetoxicosis was confirmed or suspected as a causal factor. In contrast, the current northern GoM UME has lasted more than 48 mo and has had more than 1000 reported mortalities within the currently defined spatial and temporal boundaries of the event. Initial results from the current UME do not support either morbillivirus or brevetoxin as primary causes of this event. This review is the first summary of cetacean UMEs in the GoM and provides evidence that the most common causes of previous UMEs are unlikely to be associated with the current UME.

Enhancingpopulations ofsuspension feedingbivalves,particularlythe easternoyster,Crassostreavirginica, has been proposed as a means of mitigating eutrophication in coastal waters. Review of studies evaluating the effects of C. virginica on nitrogen (N) cycling found that oysters can have effects on water quality that vary by orders of magnitude among sites, seasons, and growing condition (e.g., oyster reefs, aquaculture). Nitrogen contained in phytoplankton consumed by oysters may be returned to the water column, assimilatedintooystertissue and shell,buriedin the sediments,orreturnedtothe atmosphereas dinitrogen gas, primarily via denitrification. Accurately quantifying oyster-related N removal requires detailed knowledge oftheseprimaryfatesofNincoastalwaters.Areviewofexistingdatademonstratedthatthecurrentstateof knowledgeisincompleteinmanyrespects.Nitrogenassimilatedintooystertissueandshellpergramofdry weight was generallysimilaracrosssitesand in oysters growingon reefscomparedto aquaculture.Data on long-term burial of N associated with oyster reefs or aquaculture are lacking. When compared to suitable reference sites, denitrification rates were not consistently enhanced. Depending on environmental and oyster growing conditions, changes in denitrification rates varied by orders of magnitude among studies and did not always occur. Oyster aquaculture rarely enhanced denitrification. Unharvested oyster reefs frequently enhanced denitrification rates. Incorporating oysters into nutrient reduction strategies will require filling gaps in existing data to determine the extent to which relationships between N removal and environmental and/or growing conditions can be generalized.

Predation of fish on their scyphozoan hosts has not been clearly defined using analysis of gut contents because gelatinous prey are difficult to visually detect and are dissolved by fixative solutions. Therefore, scyphomedusae have been generally considered not relevant in fish diet. To determine the contribution of their scyphozoan host tissue to the assimilated diet of age-0 Chloroscombrus chrysurus, we determined δ13C and δ15N of fish, their hosts (scyphomedusae Aurelia sp. and Drymonema larsoni) and their potential prey—small plankton ( 200 μm)—in the coastal waters of Alabama, USA. The diet of C. chrysurus was defined using the Bayesian mixing model Stable Isotope Analysis in R (SIAR). Models indicated that the scyphozoan hosts contributed on average ~90 % to fish assimilated diet. In contrast with previous dietary assessments based on analysis of gut contents, these results highlight that scyphozoans are important to the diet of fish associated with them. Because several ecologically and economically important fish species live in association with scyphomedusae, a redefinition of trophic links in marine food webs may be needed in light of the findings in this study.

Application of stable isotope analysis (SIA) in jellyfish allows definition of trophic patterns not detectable using gut content analysis alone, but analytical protocols require standardization to avoid bias in interpreting isotopic data. We determined δ13C and δ15N in Aurelia sp. from the northern Gulf of Mexico (30°00′N, 89°00′W–30°24′N, 88°00′W) to define differences in stable isotope composition between body parts and whole body, the effect of lipid extraction on δ13C in tissues, and fractionation values from medusa to prey. The isotopic composition of bell and whole Aurelia sp. was not different. The increase in δ13C values after lipid removal suggested a correction is needed. To aid future analyses, we derived a correction equation from empirical data for jellyfish samples. Laboratory feeding experiments indicated medusae increased +4 ‰ in δ13C and +0.1 ‰ in δ15N compared to their diet. These results suggest protocols commonly applied for other species may be inaccurate to define Aurelia sp. trophic ecology. Because Aurelia spp. are commonly found in marine ecosystems, accurately defining their trophic role by use of SIA has implications for understanding marine food webs worldwide.

Veliko Jezero (Mljet, Croatia) is a nearly enclosed karstic depression filled with saltwater, where jellyfish and prey exchange with the Adriatic Sea is negligible, making this small ecosystem ideal for the controlled study of medusae in food webs. Based on the analysis of their gut contents, medusae appeared to ingest less carbon than expected on the basis of their carbon content. To accurately define carbon ingestion by medusae, we determined the diet of Aurelia sp. 5 (Dawson and Jacobs, 2001) (Scyphozoa: Semaeostomeae) from Veliko Jezero by combining gut content and stable isotope analyses. During daytime, gut contents identified a mix of small copepods as the dominant prey (62%). In contrast, feeding models based on stable isotope values of medusae and their potential prey analyzed using Stable Isotope Analysis in R indicated appendicularians could have made the greatest contribution to Aurelia sp. 5 diet (14–78%), followed by calanoid copepods (0–50%) and fish larvae (0–43%). Because appendicularians and fish larvae are abundant with medusae at night near the bottom of the “lake” and contain more carbon than small copepods, we suggest diel movements of Aurelia sp. 5 and carbon content of prey determine the carbon assimilated by medusae, with night-time prey making a

Narragansett Bay, Rhode Island, has been heavily influenced by anthropogenic nutrients for more than 200 years. Recent efforts to improve water quality have cut sewage nitrogen (N) loads to this point source estuary by more than half. Given that the bay has been heavily fertilized for longer than monitoring programs have been in place, we sought additional insight into how N dynamics in the system have historically changed. To do this, we measured the N stable isotope (δ15N) values in clam shells from as early as 3000 BP to the present. Samples from Native American middens were compared with those collected locally from museums, an archeological company, and graduate student thesis projects, during a range of time periods. Overall, δ15N values in clam shells from Narragansett Bay have increased significantly over time, reflecting known patterns of anthropogenic nutrient enrichment. Pre-colonization midden shell δ15N values were significantly lower than those post-European contact. While there were no statistical differences among shells dated from the late fifteenth century to 2005, there was a significant difference between 2005 and 2015 shells, which we attribute to the higher δ15N values in the effluent associated with recent sewage treatment upgrades. In contrast, the δ15N values of shells from the southern Rhode Island coast remained constant through time; while influenced by human activities, these areas are not directly influenced by point-source sewage discharge. Overall, our results show that this isotope technique for measuring δ15N values in clam shells provides useful insight into how N dynamics in coastal ecosystems have changed during thousands of years, providing managers vital historical information when setting goals for N reduction.

An unusual mortality event (UME) involving primarily common bottlenose dolphins Tursiops truncatus of all size classes stranding along coastal Louisiana, Mississippi, and Alabama, USA, started in early 2010 and continued into 2014. During this northern Gulf of Mexico UME, a distinct cluster of perinatal dolphins (total body length

Threats to wild horseshoe crab populations and growing interest in their use for research, education and biomedical applications have prompted demand for improved techniques to rear and maintain crabs in captivity. Although numerous laboratory studies have been conducted to determine growth and survival of horseshoe crabs under various conditions, these data have not been compiled and summarized to inform culture practices. We surveyed the literature and analysed the range of available techniques to identify and define a consistent set of conditions for maximum growth and survival of horseshoe crabs in culture. We considered three age classes; embryo, juvenile and adult, and included all extant species (Limulus polyphemus, Carcinoscorpius rotundicauda, Tachypleus gigas and Tachypleus tridentatus). We discovered relatively few published studies that clearly related husbandry conditions to growth and survival. Comparison among studies was complicated by inconsistent data collection and reporting techniques. Most published sources reported data for younger age classes, and more studies considered L. polyphemus than Asian species. The most commonly reported variables (temperature, salinity, enclosure maintenance and diet composition) showed size dependent and in some cases species-specific effects on growth and survival that will be important in guiding culture efforts. We suggest that future studies give additional consideration to substrate type, water flow, dissolved oxygen concentrations, diet quality and the quantity and frequency of feeding. If laboratory-reared stocks are to be used for propagation and restoration activities, future studies will benefit from closing these data gaps and promoting international data sharing.

Quantifying the trophic role of sharks in coastal ecosystems is crucial for the construction of accurate ecosystem models. This is particularly important for wide-ranging species like the Atlantic sharpnose shark (Rhizoprionodon terraenovae), ubiquitous across the northern Gulf of Mexico. We used gut content and stable isotope analyses to determine if differences in abundance of Atlantic sharpnose sharks in the waters around Mobile Bay, Alabama translated into differences in dietary sources or trophic position among sharks sampled east and west relative to the mouth of the bay. Gut content analysis suggested that Atlantic sharpnose sharks eat primarily teleost fishes (%IRI > 90% across size classes), and both stomach content and stable isotope analyses highlighted an ontogenetic shift in diet. Nitrogen stable isotope data from liver and muscle tissues indicated regional shifts in trophic position for Atlantic sharpnose sharks. The mixing model SIAR (stable isotope analysis in R) v.4.0.2 was used to suggest possible contributions from likely prey items for Atlantic sharpnose sharks sampled east and west of Mobile Bay. Portunid crabs and shrimp made higher contributions to the diet of Atlantic sharpnose sharks in the western region, compared to higher and more variable contributions from fish like croaker (Micropogonias undulatus) and hardhead catfish (Arius felis) in the eastern region. Our results suggest trophic plasticity in Atlantic sharpnose sharks, findings that emphasize the importance of examining regional variation in trophic position when constructing coastal foodweb models.

We measured seasonal effects of wastewater treatment plant (WTP) effluent on growth, survival, and accumulation of microbes in oysters near a major WTP in Mobile Bay, AL. Despite higher nutrients near the WTP, seasonal conditions rather than distance affected chloro- phyll a concentration and oyster growth. In summer and fall, when oyster growth was higher, δ 15 N‰ in oysters near the WTP changed through time to reflect δ 15 N‰ in effluent (approx. −4‰). Microbial indicators (male-specific coli- phage, fecal coliforms) were highest in oysters near the WTP in all seasons and correlated with δ 15 N‰ in fall and summer. Increased riverine discharge and slower acquisition of δ 15 N‰ likely confounded correlations in winter/spring.