Reef structure of the Florida Reef Tract for the period 2005-2020.

Shallow-water coral reefs of the Florida Reef Tract compose the third largest reef in the world, but during the last several decades, scleractinian (stony) corals have suffered unprecedented declines from global and local stressors. A program to evaluate the effects of high-temperature bleaching events was initiated by The Nature Conservancy's Florida Reef Resilience Program in 2005 and surveys have been completed across at least some portion of the entire region every year since. The program adopted a demographic (colony-based) assessment approach, which records colony species, size (height and maximum diameter), and estimated partial mortality (percent barren skeleton). Because reef structure is critical to ecosystem functioning and services, data from 2005 to 2020 were analyzed to describe the abundance, size, and morphological complexity of stony coral colonies forming the biogenic reef. Colony height, footprint, surface area, and volume summed for 6016 transects were used to describe reef structure and averages were used to characterize the components that contributed to the structure. Nearly 150,000 colonies representing 49 species were reported during this period and results demonstrated both spatial and temporal changes for the region and for geographic subregions. Some subregions showed increasing colony density, especially for three small, hemispheric species, and declining average colony size.

Skeletal growth capacity as a measure of coral species and community resilience.

Accretion and erosion of scleractinian (stony coral) carbonate skeletons determine whether a colony will increase or decrease in size with potential consequences for ecosystem processes, functions and services. The capacity for skeletal growth can be estimated by comparing a colony's rate of calcification with its rate of erosion. Calcification depends on the species-specific metabolic activity of living tissue, and erosion depends primarily on the availability and density of barren skeleton, those areas on the colony where polyps have died. Assessment of skeletal growth capacity requires data on calcification rates, erosion rates and both live and barren colony surface area. Rates of calcification and erosion are documented for many Caribbean species and others can be estimated from existing data. Three-dimensional surface area of colonies can be determined from data collected during demographic surveys, which identify species, measure dimensions, and estimate the proportion of live tissue on a colony. Data from demographic surveys conducted in the U.S. Virgin Islands are used to calculate the skeletal growth capacity (GC) as an indicator of coral species and community resilience. Scleractinia are the primary architects of coral reefs, and the gain or loss of skeletal framework is vitally important to reef ecosystem processes that lead to valued goods and services. Estimates of GC reflect stony coral resilience, which is the capacity to recover from disturbances by returning to previous physical and functional levels. GC can also provide insight to the effects of stressors such as ocean acidification, and can inform several management decisions, including restoration site selection and threatened species designation.

A biological condition gradient for Caribbean coral reefs: Part II. Numeric rules using sessile benthic organisms.

The Biological Condition Gradient (BCG) is a conceptual model used to describe incremental changes in biological condition along a gradient of increasing anthropogenic stress. As coral reefs collapse globally, scientists and managers are focused on how to sustain the crucial structure and functions, and the benefits that healthy coral reef ecosystems provide for many economies and societies. We developed a numeric (quantitative) BGC model for the coral reefs of Puerto Rico and the US Virgin Islands to transparently facilitate ecologically meaningful management decisions regarding these fragile resources. Here, reef conditions range from natural, undisturbed conditions to severely altered or degraded conditions. Numeric decision rules were developed by an expert panel for scleractinian corals and other benthic assemblages using multiple attributes to apply in shallow-water tropical fore reefs with depths <30 m. The numeric model employed decision rules based on metrics (e.g., % live coral cover, coral species richness, pollution-sensitive coral species, unproductive and sediment substrates, % cover by Orbicella spp.) used to assess coral reef condition. Model confirmation showed the numeric BCG model predicted the panel's median site ratings for 84% of the sites used to calibrate the model and 89% of independent validation sites. The numeric BCG model is suitable for adaptive management applications and supports bioassessment and criteria development. It is a robust assessment tool that could be used to establish ecosystem condition that would aid resource managers in evaluating and communicating current or changing conditions, protect water and habitat quality in areas of high biological integrity, or develop restoration goals with stakeholders and other public beneficiaries.

A biological condition gradient for coral reefs in the US Caribbean Territories: Part I. Coral narrative rules.

As coral reef condition and sustainability continue to decline worldwide, losses of critical habitat and their ecosystem services have generated an urgency to understand and communicate reef response to management actions, environmental contamination, and natural disasters. Increasingly, coral reef protection and restoration programs emphasize the need for robust assessment tools for protecting high-quality waters and establishing conservation goals. Of equal importance is the need to communicate assessment results to stakeholders, beneficiaries, and the public so that environmental consequences of decisions are understood. The Biological Condition (BCG) model provides a structure to evaluate the condition of a coral reef in increments of change along a gradient of human disturbance. Communication of incremental change, regardless of direction, is important for decision makers and the public to better understand what is gained or lost depending on what actions are taken. We developed a narrative (qualitative) Biological Condition Gradient (BCG) from the consensus of a diverse expert panel to provide a framework for coral reefs in US Caribbean Territories. The model uses narrative descriptions of biological attributes for benthic organisms to evaluate reefs relative to undisturbed or minimally disturbed conditions. Using expert elicitation, narrative decision rules were proposed and deliberated to discriminate among six levels of change along a gradient of increasing anthropogenic stress. Narrative rules for each of the BCG levels are presented to facilitate the evaluation of benthic communities in coral reefs and provide specific narrative features to detect changes in coral reef condition and biological integrity. The BCG model can be used in the absence of numeric, or quantitative metrics, to evaluate actions that may encroach on coral reef ecosystems, manage endangered species habitat, and develop and implement management plans for marine protected areas, watersheds, and coastal zones. The narrative BCG model is a defensible model and communication tool that translates scientific results so the nontechnical person can understand and support both regulatory and non-regulatory water quality and natural resource programs.

Biological Assessment of Coral Reefs in Southern Puerto Rico: A Technical Approach for Coral Reef Protection Under the U.S. Clean Water Act.

States and other jurisdictions may protect coral reefs using biological water quality standards outlined by the United States Clean Water Act (CWA). Such protection will require long-term, regional monitoring of the resource using biological indicators and a probability-based sampling design. A 60-station survey targeting nearshore linear coral reef was conducted across southern Puerto Rico in December 2011 to document the status of reef inhabitants using a probabilistic, regional sampling design. The quantity, type and condition of stony corals, fish, gorgonians and sponges were documented from each station, providing a robust representation of linear reef status and composition across the region. Fish represented 106 unique taxa and stony corals 32 unique taxa. Benthic organisms (stony corals, sponges and gorgonians) averaged nearly 12 colonies per square meter, more than half of which were gorgonians. Assessment results can be used as a baseline to compare with future regional surveys to quantify change in reef condition over time (trend). Both temporal and spatial changes can be expected after large-scale disturbances like hurricanes Maria and Irma in 2017. The indicators and probabilistic sampling design support the long-term regional monitoring envisioned by the Environmental Protection Agency to implement CWA protections in Puerto Rico and elsewhere.

An introduction to Bayesian networks as assessment and decision support tools for managing coral reef ecosystem services.

Quantifying ecosystem goods and services can help evaluate policies aimed at protecting present and future generations from losing ecosystem benefits. Explicating and quantifying the relationships among risk factors, ecological structure and function, and delivery of ecosystem goods and services requires analytical methodologies that propagate uncertainties. The capabilities of Bayesian networks in generating predictions and accounting for uncertainty are explored with a focus on coral reef ecosystem service assessments. The qualitative aspects of Bayesian networks can be applied to conceptual frameworks developed for coral reef ecosystem service assessments. This is demonstrated using qualitative graphs that describe the relationships between coral reef condition endpoints and benefits from ecosystem services including property protection, recreational opportunities, fish for fisheries, and biochemical metabolites for commercial products developed from reef organisms. Bayesian networks help weigh uncertainties between management decision impacts on stressors and the corresponding delivery of ecosystem services. Quantitative capabilities for inferences are examined in hypothetical scenarios evaluating how decisions affect coral reef ecosystem services and economic benefits and resilience to episodic stress. The described methods facilitate visualizing the potential impacts on ecosystem services from alternative scenarios.

Spatial Distribution of Collections Yielding Marine Natural Products.

The societal benefits of coral reef ecosystems include shoreline protection, habitat provision for reef fish, tourism, and recreation. Rarely considered in valuation of reefs is the considerable contribution of marine natural products (MNPs) to both human health and the economy. To better understand the relation of MNP discovery with the characteristics and condition of coral reef ecosystems, we initiated a study to track the collection location and taxonomic identity of organisms that have provided pharmacological products. We reviewed collection information and associated data from 298 pharmacological products originating from marine biota during the past 47 years. The products were developed from 232 different marine species representing 15 phyla, and the 1296 collections of these specimens occurred across 69 countries and seven continents. Our evaluation of the collection data was hampered by sundry observational and reporting issues, including imprecise location descriptions and omission of collection dates. Nonetheless, the study provides an important synopsis and appraisal of years of study and exploration by the marine natural product community. Understanding and quantifying the benefits of MNP discovery will depend upon improved reporting of collections, including accurate taxonomic identification, collection dates, and locations.

Calcification in Caribbean reef-building corals at high pCO2 levels in a recirculating ocean acidification exposure system.

Projected increases in ocean pCO2 levels are anticipated to affect calcifying organisms more rapidly and to a greater extent than other marine organisms. The effects of ocean acidification (OA) have been documented in numerous species of corals in laboratory studies, largely tested using flow-through exposure systems. We developed a recirculating ocean acidification exposure system that allows precise pCO2 control using a combination of off-gassing measures including aeration, water retention devices, venturi injectors, and CO2 scrubbing. We evaluated the recirculating system performance in off-gassing effectiveness and maintenance of target pCO2 levels over an 84-day experiment. The system was used to identify changes in calcification and tissue growth in response to elevated pCO2 (1000 µatm) in three reef-building corals of the Caribbean: Pseudodiploria clivosa, Montastraea cavernosa, and Orbicella faveolata. All three species displayed an overall increase in net calcification over the 84-day exposure period regardless of pCO2 level (control +0.28- 1.12 g, elevated pCO2 +0.18- 1.16 g), and the system was effective at both off-gassing acidified water to ambient pCO2 levels, and maintaining target elevated pCO2 levels over the 3-month experiment.

Linking ecosystem service supply to stakeholder concerns on both land and sea: An example from Guánica Bay watershed, Puerto Rico.

Policies to protect coastal resources may lead to greater social, economic, and ecological returns when they consider potential co-benefits and trade-offs on land. In Guánica Bay watershed, Puerto Rico, a watershed management plan is being implemented to restore declining quality of coral reefs due to sediment and nutrient runoff. However, recent stakeholder workshops indicated uncertainty about benefits for the local community. A total of 19 metrics were identified to capture stakeholder concerns, including 15 terrestrial ecosystem services in the watershed and 4 metrics in the coastal zone. Ecosystem service production functions were applied to quantify and map ecosystem service supply in 1) the Guánica Bay watershed and 2) a highly engineered upper multi-watershed area connected to the lower watershed via a series of reservoirs and tunnels. These two watersheds were compared to other watersheds in Puerto Rico. Relative to other watersheds, the Upper Guánica watershed had high air pollutant removal rates, forest habitat area, biodiversity of charismatic and endangered species, but low farmland quality and low sediment retention. The Lower Guánica watershed had high rates of denitrification and high levels of marine-based recreational and fishing opportunities compared to other watersheds, but moderate to low air pollutant removal, soil carbon content, sediment and nutrient retention, and terrestrial biodiversity. Our results suggest that actions in the watershed to protect coral reefs may lead to improvements in other ecosystem services that stakeholders care about on land. Considering benefits from both coastal and terrestrial ecosystems in making coastal management decisions may ultimately lead to a greater return on investment and greater stakeholder acceptance, while still achieving conservation goals.

Regional status assessment of stony corals in the US Virgin Islands.

States may protect coral reefs using biological water quality standards outlined by the Clean Water Act. This requires biological assessments with indicators sensitive to human disturbance and regional, probability-based survey designs. Stony coral condition was characterized on a regional scale for the first time in the nearshore waters of the US Virgin Islands (USVI). Coral composition, abundance, size, and health were assessed at 66 stations in the St. Croix region in fall 2007 and at 63 stations in the St. Thomas and St. John region in winter 2009. Indicators were chosen for their sensitivity to human disturbance. Both surveys were probability-based (random) designs with station locations preselected from areas covered by hardbottom and coral reef substrate. Taxa richness was as high as 21 species but more than half the area of both regions exhibited taxa richness of <10 species in the 25 m(2) transect area. Coral density was as high as 5 colonies m(-2) but more than half the area of both regions had <2 colonies m(-2). Both regions showed similar dominant species based on frequency of occurrence and relative abundance. Because of large colony sizes, Montastrea annularis provided more total surface area and live surface area than more abundant species. The surveys establish baseline regional conditions and provide a foundation for long-term regional monitoring envisioned by the USVI Department of Planning and Natural Resources. The probabilistic sampling design assures the data can be used in Clean Water Act reporting.

A decision support framework for science-based, multi-stakeholder deliberation: a coral reef example.

We present a decision support framework for science-based assessment and multi-stakeholder deliberation. The framework consists of two parts: a DPSIR (Drivers-Pressures-States-Impacts-Responses) analysis to identify the important causal relationships among anthropogenic environmental stressors, processes, and outcomes; and a Decision Landscape analysis to depict the legal, social, and institutional dimensions of environmental decisions. The Decision Landscape incorporates interactions among government agencies, regulated businesses, non-government organizations, and other stakeholders. It also identifies where scientific information regarding environmental processes is collected and transmitted to improve knowledge about elements of the DPSIR and to improve the scientific basis for decisions. Our application of the decision support framework to coral reef protection and restoration in the Florida Keys focusing on anthropogenic stressors, such as wastewater, proved to be successful and offered several insights. Using information from a management plan, it was possible to capture the current state of the science with a DPSIR analysis as well as important decision options, decision makers and applicable laws with a the Decision Landscape analysis. A structured elicitation of values and beliefs conducted at a coral reef management workshop held in Key West, Florida provided a diversity of opinion and also indicated a prioritization of several environmental stressors affecting coral reef health. The integrated DPSIR/Decision landscape framework for the Florida Keys developed based on the elicited opinion and the DPSIR analysis can be used to inform management decisions, to reveal the role that further scientific information and research might play to populate the framework, and to facilitate better-informed agreement among participants.

Exosomes are secreted at similar densities by M21 and PC3 human cancer cells and show paclitaxel solubility.

Exosomes are cell-secreted vesicles less than ≈150 nm in size that contain gene-encoding and gene-silencing RNA and cytosolic proteins with roles in intercellular communication. Interest in the use of exosomes as targeted drug delivery vehicles has grown since it was shown that they can bind specific cells and deliver intact genetic material to the cytosol of target cells. We isolated extracellular vesicles (EVs), consisting of a mixture of exosomes and microvesicles, from prostate (PC3) and melanoma (M21) cancer cell lines using serial ultracentrifugation. Interrogation via western blot analysis confirmed enrichment of CD63, a widely recognized EV surface protein, in the EV pellet from both cell lines. Nanoparticle tracking analysis (NTA) of EV pellets revealed that the two cell lines produced distinct vesicle size profiles in the ≈30 nm to ≈400 nm range. NTA further showed that the fraction of exosomes to all EVs was constant, suggesting cellular mechanisms that control the fraction of secreted vesicles that are exosomes. Transmission electron microscopy (TEM) images of the unmodified PC3 EVs showed vesicles with cup-like (i.e., nanocapsule) and previously unreported prolate morphologies. The observed non-spherical morphologies for dehydrated exosomal vesicles (size ≈30-100 nm) are most likely related to the dense packing of proteins in exosome membranes. Solubility phase diagram data showed that EVs enhanced the solubility of paclitaxel (PTX) in aqueous solution compared to a water-only control. Combined with their inherent targeting and cytosol delivery properties, these findings highlight the potential advantages of using exosomes as chemotherapeutic drug carriers in vivo.

Paclitaxel-Loaded Cationic Fluid Lipid Nanodiscs and Liposomes with Brush-Conformation PEG Chains Penetrate Breast Tumors and Trigger Caspase-3 Activation.

Novel approaches are required to address the urgent need to develop lipid-based carriers of paclitaxel (PTX) and other hydrophobic drugs for cancer chemotherapy. Carriers based on cationic liposomes (CLs) with fluid (i.e., chain-melted) membranes (e.g., EndoTAG-1) have shown promise in preclinical and late-stage clinical studies. Recent work found that the addition of a cone-shaped poly(ethylene glycol)-lipid (PEG-lipid) to PTX-loaded CLs (CLsPTX) promotes a transition to sterically stabilized, higher-curvature (smaller) nanoparticles consisting of a mixture of PEGylated CLsPTX and PTX-containing fluid lipid nanodiscs (nanodiscsPTX). These CLsPTX and nanodiscsPTX show significantly improved uptake and cytotoxicity in cultured human cancer cells at PEG coverage in the brush regime (10 mol % PEG-lipid). Here, we studied the PTX loading, in vivo circulation half-life, and biodistribution of systemically administered CLsPTX and nanodiscsPTX and assessed their ability to induce apoptosis in triple-negative breast-cancer-bearing immunocompetent mice. We focused on fluid rather than solid lipid nanodiscs because of the significantly higher solubility of PTX in fluid membranes. At 5 and 10 mol % of a PEG-lipid (PEG5K-lipid, molecular weight of PEG 5000 g/mol), the mixture of PEGylated CLsPTX and nanodiscsPTX was able to incorporate up to 2.5 mol % PTX without crystallization for at least 20 h. Remarkably, compared to preparations containing 2 and 5 mol % PEG5K-lipid (with the PEG chains in the mushroom regime), the particles at 10 mol % (with PEG chains in the brush regime) showed significantly higher blood half-life, tumor penetration, and proapoptotic activity. Our study suggests that increasing the PEG coverage of CL-based drug nanoformulations can improve their pharmacokinetics and therapeutic efficacy.

Assessing Coral Reef Condition Indicators for Local and Global Stressors Using Bayesian Networks.

Coral reefs are highly valued ecosystems currently threatened by both local and global stressors. Given the importance of coral reef ecosystems, a Bayesian network approach can benefit an evaluation of threats to reef condition. To this end, we used data to evaluate the overlap between local stressors (overfishing and destructive fishing, watershed-based pollution, marine-based pollution, and coastal development threats), global stressors (acidification and thermal stress), and management effectiveness with indicators of coral reef health (live coral index, live coral cover, population bleaching, colony bleaching, and recently killed corals). Each of the coral health indicators had Bayesian networks constructed globally and for Pacific, Atlantic, Australia, Middle East, Indian Ocean, and Southeast Asia coral reef locations. Sensitivity analysis helped evaluate the strength of the relationships between different stressors and reef condition indicators. The relationships between indicators and stressors were also evaluated with conditional analyses of linear and nonlinear interactions. In this process, a standardized direct effects analysis was emphasized with a target mean analysis to predict changes in the mean value of the reef indicator from individual changes to the distribution of the predictor variables. The standardized direct effects analysis identified higher risks in the Middle East for watershed-based pollution with population bleaching and in Australia for overfishing and destructive fishing with living coral. For thermal stress, colony bleaching and recently killed coral in the Indian Ocean were found to have the strongest direct associations along with living coral in the Middle East. For acidification threat, Australia had a relatively strong association with colony bleaching, and the Middle East had the strongest overall association with recently killed coral, although extrapolated spatial data were used for the acidification estimates. The Bayesian network approach helped to explore the relationships among existing databases used for policy development in coral reef management by examining the sensitivity of multiple indicators of reef condition to spatially distributed stress. Integr Environ Assess Manag 2021;17:165-187. Published 2020. This article is a US Government work and is in the public domain in the USA.

Paclitaxel loading in cationic liposome vectors is enhanced by replacement of oleoyl with linoleoyl tails with distinct lipid shapes.

Lipid carriers of hydrophobic paclitaxel (PTX) are used in clinical trials for cancer chemotherapy. Improving their loading capacity requires enhanced PTX solubilization. We compared the time-dependence of PTX membrane solubility as a function of PTX content in cationic liposomes (CLs) with lipid tails containing one (oleoyl; DOPC/DOTAP) or two (linoleoyl; DLinPC/newly synthesized DLinTAP) cis double bonds by using microscopy to generate kinetic phase diagrams. The DLin lipids displayed significantly increased PTX membrane solubility over DO lipids. Remarkably, 8 mol% PTX in DLinTAP/DLinPC CLs remained soluble for approximately as long as 3 mol% PTX (the solubility limit, which has been the focus of most previous studies and clinical trials) in DOTAP/DOPC CLs. The increase in solubility is likely caused by enhanced molecular affinity between lipid tails and PTX, rather than by the transition in membrane structure from bilayers to inverse cylindrical micelles observed with small-angle X-ray scattering. Importantly, the efficacy of PTX-loaded CLs against prostate cancer cells (their IC50 of PTX cytotoxicity) was unaffected by changing the lipid tails, and toxicity of the CL carrier was negligible. Moreover, efficacy was approximately doubled against melanoma cells for PTX-loaded DLinTAP/DLinPC over DOTAP/DOPC CLs. Our findings demonstrate the potential of chemical modifications of the lipid tails to increase the PTX membrane loading while maintaining (and in some cases even increasing) the efficacy of CLs. The increased PTX solubility will aid the development of liposomal PTX carriers that require significantly less lipid to deliver a given amount of PTX, reducing side effects and costs.

Acidification in the U.S. Southeast: Causes, Potential Consequences and the Role of the Southeast Ocean and Coastal Acidification Network.

Coastal acidification in southeastern U.S. estuaries and coastal waters is influenced by biological activity, run-off from the land, and increasing carbon dioxide in the atmosphere. Acidification can negatively impact coastal resources such as shellfish, finfish, and coral reefs, and the communities that rely on them. Organismal responses for species located in the U.S. Southeast document large negative impacts of acidification, especially in larval stages. For example, the toxicity of pesticides increases under acidified conditions and the combination of acidification and low oxygen has profoundly negative influences on genes regulating oxygen consumption. In corals, the rate of calcification decreases with acidification and processes such as wound recovery, reproduction, and recruitment are negatively impacted. Minimizing the changes in global ocean chemistry will ultimately depend on the reduction of carbon dioxide emissions, but adaptation to these changes and mitigation of the local stressors that exacerbate global acidification can be addressed locally. The evolution of our knowledge of acidification, from basic understanding of the problem to the emergence of applied research and monitoring, has been facilitated by the development of regional Coastal Acidification Networks (CANs) across the United States. This synthesis is a product of the Southeast Coastal and Ocean Acidification Network (SOCAN). SOCAN was established to better understand acidification in the coastal waters of the U.S. Southeast and to foster communication among scientists, resource managers, businesses, and governments in the region. Here we review acidification issues in the U.S. Southeast, including the regional mechanisms of acidification and their potential impacts on biological resources and coastal communities. We recommend research and monitoring priorities and discuss the role SOCAN has in advancing acidification research and mitigation of and adaptation to these changes.

Development and implementation of coral reef biocriteria in U.S. jurisdictions.

Coral reefs worldwide are declining at an alarming rate and are under continuous threat from both natural and anthropogenic environmental stressors. Warmer sea temperatures attributed to global climate change and numerous human activities at local scales place these valuable ecosystems at risk. Reefs provide numerous services, including shoreline protection, fishing, tourism and biological diversity, which are lost through physical damage, overfishing, and pollution. Pollution can be controlled under provisions of the Clean Water Act, but these options have not been fully employed to protect coral reefs. No U.S. jurisdiction has implemented coral reef biocriteria, which are narrative or quantitative water quality standards based on the condition of a biological resource or assemblage. The President's Ocean Action Plan directs the U.S. Environmental Protection Agency (EPA) to develop biological assessment methods and biological criteria for evaluating and maintaining the health of coral reef ecosystems. EPA has formed the Coral Reef Biocriteria Working Group (CRBWG) to foster development of coral reef biocriteria through focused research, evaluation and communication among Agency partners and U.S. jurisdictions. Ongoing CRBWG activities include development and evaluation of a rapid bioassessment protocol for application in biocriteria programs; development of a survey design and monitoring strategy for the U.S. Virgin Islands; comprehensive reviews of biocriteria approaches proposed by states and territories; and assembly of data from a variety of monitoring programs for additional metrics. Guidance documents are being prepared to assist U.S. jurisdictions in reaching protective and defensible biocriteria.

Evaluation of stony coral indicators for coral reef management.

Colonies of reef-building stony corals at 57 stations around St. Croix, US Virgin Islands were characterized by species, size and percentage of living tissue. Taxonomic, biological and physical indicators of coral condition were derived from these measurements and assessed for their response to gradients of human disturbance-a requirement for indicators used in regulatory assessments under authority of the Clean Water Act. At the most intensely disturbed location, five of eight primary indicators were highly correlated with distance from the source of disturbance: Coral taxa richness, average colony size, the coefficient of variation of colony size, total topographic coral surface area, and live coral surface area. An additional set of exploratory indicators related to rarity, reproductive and spawning mode and taxonomic identity were also screened. The primary indicators demonstrated sufficient precision to detect levels of change that would be applicable in a regional-scale regulatory program.

IDENTIFYING AND STRUCTURING OBJECTIVES FOR A CORAL REEF PROTECTION PLAN AT THE U.S. ENVIRONMENTAL PROTECTION AGENCY.

Region 2 of the U.S. Environmental Protection Agency initiated a Coral Reef Protection Plan (CRPP) in 2014 to reduce anthropogenic stress on Caribbean coral reefs. The CRPP is intended to foster institutional practices that improve reef condition and focus regulatory and nonregulatory decision making on minimizing pollutant release to coastal systems. A framework incorporating two sets of objectives was constructed to examine the short- and long-term costs and benefits of tasks. The first set of objectives was derived from existing tasks in the CRPP and was intended to support an update of the CRPP for 2015. Fundamental objectives were constructed to communicate the end objectives across tasks and means objectives were constructed to communicate the means for achieving them. The second set of objectives was created to reflect costs and benefits of the CRPP beyond 2015. These objectives contained fundamental objectives comprising broad social, economic, learning and governance topics. The means objectives included tasks such as building capacity, providing regulatory oversight, and learning and reducing uncertainties. The second set of objectives also included strategic objectives that identify long-range benefits such as coral reef integrity and reef ecosystem services. The process of defining objectives helped to ascertain and better elucidate the important consequences for the CRPP. Understanding objectives not only provides a roadmap for coral reef protection but can help Region 2 communicate internally and externally with other agencies, industry, and the public.

Greater hemocyte bactericidal activity in oysters (Crassostrea virginica) from a relatively contaminated site in Pensacola Bay, Florida.

Bivalve mollusks such as Crassostrea virginica inhabiting polluted estuaries and coastal areas may bioaccumulate high concentrations of contaminants without apparent ill effects. However, changes in putative internal defense activities have been associated with contaminant accumulation in both experimental and long-term field exposures. In an effort to elucidate these relationships, 40 oysters were collected from Bayou Chico (BC) and East Bay (EB) in Pensacola Bay, FL, two estuaries known to differ in the type and magnitude of chemical contaminants present. Oyster tissue concentrations of metals, tri- and dibutyltin (TBT, DBT), polycyclic aromatic hydrocarbons (PAHs) and polychlorinated biphenyls (PCBs) were measured in individual oysters, as were hemocyte counts (HCs), hemocyte bacterial killing indices (KI), serum lysozyme (LYS) and serum protein (PRO) levels. Average HC, KI, LYS and PRO were significantly higher in BC oysters, which also had significantly higher tissue concentrations of total trace metals, butyltins (BTs), PAHs, PCBs, pesticides, and Mn, Cu, Zn and Sn. EB oysters had low organic contaminant levels and no detectable BTs, but significantly higher concentrations of Al, Cr, Fe, Ag, Cd, and Hg. Simple correlation analysis between specific defense measurements and specific chemical analytes showed specific positive relationships that corroborated previous findings in other FL estuaries. Canonical correlation analysis was used to examine relationships between defense measurements and tissue metals using linearly combined sets of variables. Results were also consistent with previous findings-the highest possible canonical correlation was positive: r=0.864, P<0.0019 among canonical variables composed of HC, KI and LYS for defense, and Fe, Cu, Ag, Cd, Sb, Sn, Ni, Pb and Hg for metals.