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Diseases of marine organisms, including sponges on coral reefs, are being reported with increasing frequency worldwide. Aplysina Red Band Syndrome (ARBS) occurs across the Caribbean basin, predominantly affecting Aplysina cauliformis, one of the most common reef sponges in this region. Existing ARBS lesions and their effects on the sponge holobiont have been documented, yet little is known about the biochemical and microbial changes associated with the onset of infection. Due to the transmissible nature of ARBS, infection can be induced and monitored through sponge-to-sponge direct contact. Nine-day contact experiments with healthy-diseased and healthy-healthy sponge pairings were conducted in the Bahamas in January and July to compare individual sponges sampled initially and at one of three successive time points. Temporal changes in bacterial assemblages and photosymbiont abundance (via concentrations of chlorophyll a), and concentrations of total protein, heat shock protein 70, and major secondary metabolites that may correspond with disease onset were characterized. All healthy sponges in contact with diseased sponges developed ARBS by day 9 in January and by day 6 in July, suggesting that observed changes in the holobiont corresponded with the development of ARBS. The concentrations of several major secondary metabolites, as well as heat shock protein 70 and chlorophyll a, changed significantly in samples of visibly healthy tissue from initially healthy sponges that became diseased. In contrast, the composition of the associated bacterial community changed in all attached sponges over time. These results suggest that infection with ARBS elicits rapid responses by the sponge holobiont, providing a model system in which to investigate immune responses in an early metazoan.

Oyster reefs are vital to estuarine health, but they experience multiple stressors and globally declining populations. This study examined effects of hypoxia and tributyltin (TBT) on adult Eastern oysters (Crassostrea virginica) exposed either in the laboratory or the field following a natural hypoxic event. In the laboratory, oysters were exposed to either hypoxia followed by a recovery period, or to hypoxia combined with TBT. mRNA expression of HIF1-α and Tβ-4 along with hemocyte counts, biomarkers of hypoxic stress and immune health, respectively, were measured. In field-deployed oysters, HIF1-α and Tβ-4 expression increased, while no effect on hemocytes was observed. In contrast, after 6 and 8 days of laboratory-based hypoxia exposure, both Tβ-4 expression and hemocyte counts declined. After 8 days of exposure to hypoxia + TBT, oysters substantially up-regulated HIF1-α and down-regulated Tβ-4, although hemocyte counts were unaffected. Results suggest that hypoxic exposure induces immunosuppression which could increase vulnerability to pathogens.

Aplysina cauliformis, the Caribbean purple rope sponge, is commonly affected by Aplysina Red Band Syndrome (ARBS). This transmissible disease manifests as circular lesions with red margins and results in bare spongin fibers. Leptolyngbya spp. appear to be responsible for the characteristic red coloration but transmission studies with a sponge-derived isolate failed to establish disease, leaving the etiology of ARBS unknown. To investigate the cause of ARBS, contact transmission experiments were performed between healthy and diseased sponges separated by filters with varying pore sizes. Transmission occurred when sponges were separated by filters with pore sizes ≥ 2.5 μm, suggesting a prokaryotic pathogen(s) but not completely eliminating eukaryotic pathogen(s). Using 16S rRNA gene sequencing methods, 38 prokaryotic taxa were significantly enriched in diseased sponges, including Leptolyngbya, whereas seven taxa were only found in some, but not all, of the ARBS-affected sponges. These results do not implicate a single taxon, but rather a suite of taxa that changed in relative abundance with disease, suggesting a polymicrobial etiology as well as dysbiosis. As a better understanding of dysbiosis is gained, changes in the composition of associated prokaryotic communities may have increasing importance for evaluating and maintaining the health of individuals and imperiled coral reef ecosystems.

Many studies have evaluated the impacts of hurricanes on coral communities, but far less is known about impacts, recovery, and resilience of sponge communities to these extreme events. In September 2017, St. Thomas, U.S. Virgin Islands, was impacted by two Category 5 hurricanes within 2 weeks: Hurricanes Irma and Maria. Such extreme events occurring in such rapid succession are virtually unprecedented. Pre-hurricane (2015, 2016) surveys of permanent transects at six sites around St. Thomas were compared with those at 10 weeks post-hurricanes (December 2017) to evaluate storm impacts on sponges and on benthic coral reef constituents. These surveys also established a baseline for evaluating future recovery. Percent cover of sponges declined by 24.9% post-hurricanes. In contrast, sponge density increased by 43.9% from 2015 to 2016 and declined slightly after the hurricanes. Overall sponge volume did not vary over time, and whereas sponge diversity was similar in 2015 and 2016, it increased post-hurricanes. Sponge morphologies were differentially affected by the hurricanes; the proportion of upright sponges declined by 36.9%, while there was a 24.4% increase in encrusting sponges. Coral and macroalgal cover did not change significantly over the sampling period, while percent cover of epilithic algae increased and non-living substrata decreased from 2015 to 2016 but did not change further post-hurricanes. At all sites, recruitment and/or regrowth of sponges was observed within 10 weeks post-hurricanes, indicating potential resilience in Caribbean sponge communities. Whether these sponge communities return to pre-hurricane conditions and how long that will take remains to be seen.

Symbiotic microbial communities of sponges serve critical functions that have shaped the evolution of reef ecosystems since their origins. Symbiont abundance varies tremendously among sponges, with many species classified as either low microbial abundance (LMA) or high microbial abundance (HMA), but the evolutionary dynamics of these symbiotic states remain unknown. This study examines the LMA/HMA dichotomy across an exhaustive sampling of Caribbean sponge biodiversity and predicts that the LMA symbiotic state is the ancestral state among sponges. Conversely, HMA symbioses, consisting of more specialized microorganisms, have evolved multiple times by recruiting similar assemblages, mostly since the rise of scleractinian-dominated reefs. Additionally, HMA symbioses show stronger signals of phylosymbiosis and cophylogeny, consistent with stronger co-evolutionary interaction in these complex holobionts. These results indicate that HMA holobionts are characterized by increased endemism, metabolic dependence and chemical defences. The selective forces driving these patterns may include the concurrent increase in dissolved organic matter in reef ecosystems or the diversification of spongivorous fishes.

Oyster reefs provide essential ecosystem services but are severely degraded worldwide. Extreme flooding events, which can be intensified by water management decisions, reduce water quality in estuaries and further threaten oyster populations. Restoration and conservation of oysters is dependent on the success of early oyster life stages. This study examined the effect of water quality stressors associated with flooding events on the growth and survival of larval and juvenile oysters ( Crassostrea virginica ). In 96-h assays, we exposed D-stage larvae to a range of dissolved oxygen, microcystin-LR, pH, and salinity concentrations. These conditions were selected based on water quality data from the Mississippi Sound during a 2019 freshwater flooding event caused by the Bonnet Carre Spillway opening. There was no negative effect of microcystin-LR or pH on early veligers at the concentrations tested, but low salinity significantly reduced shell growth, and hypoxia ( −1 O2) decreased both larval growth and survival. Post-metamorphosis juvenile oysters were exposed to the same water quality stressors for 24 days in the lab. Low DO, pH, and salinity treatments reduced juvenile change in wet weight and shell growth rates, but had no effects on survival. These laboratory-exposed juveniles were subsequently deployed into the field to assess the ability of juveniles to recover from short-term exposure to simulated flooding-associated stressors. After deployment to natural conditions in the Mississippi Sound, juvenile oysters were able to compensate for reduced growth during the lab exposure, even though survival was reduced for juveniles previously exposed to low pH during the first two weeks in the field. In general, early oyster life stages were relatively tolerant of the duration and stressor concentrations tested, but negative sublethal impacts of flood-associated stressors must be considered in the face of increasing frequency and duration of flooding events due to climate change.

As coral cover has declined on Caribbean reefs, space has become occupied by other benthic taxa, including sponges, which may affect the recruitment of new corals, thereby affecting the ability of reefs to recover to coral-dominated states. Sponges may inhibit coral recruitment by pre-empting potential recruitment space, overgrowing recruits, or through allelopathy. This study examined coral recruitment across six coral reef sites surrounding St. Thomas, US Virgin Islands, and the impact of one species of sponge and the sponge community as a whole on coral recruitment. To test the effect of a single species of sponge on coral recruitment, fragments of living or non-living Aplysina cauliformis were attached to terracotta recruitment tiles and deployed at all six sites, along with unoccupied tiles as controls. At two of the sites, a community-level experiment consisted of deploying recruitment tiles in 1 m2 plots that were either cleared of the entire sponge community or control plots where no sponges were removed. Recruitment rates showed a consistent difference among sites over multiple years and experiments. Results of the species-specific experiment showed that the proximity of live or dead A. cauliformis did not affect coral recruitment. However, results of the community-level experiment found greater coral recruitment rates in plots cleared of sponges, suggesting that the presence of the sponge community negatively affected coral recruitment. This study is one of the first to experimentally test and find a significant impact of sponges on coral recruitment, and highlights the need for additional research in this area.

Predation on corals by visual predators is a significant source of partial or total mortality on coral reefs, and corals have evolved strategies, including chemical defenses, to deter predation. One mechanism that organisms use to communicate the presence of chemical defenses is aposematic coloration, or the display of bright coloration as a warning to visual predators such as fish. Corals exhibit multiple colors, and it has been hypothesized that one role for this variability in coloration is as an aposematic warning of adverse palatability. Here, we test green and orange color morphs of the Caribbean coral Montastraea cavernosa for the presence of chemical defenses and whether their differences in coloration elicited different feeding responses. While M. cavernosa is chemically defended, there is no difference in feeding deterrence between color morphs; thus, the different color morphs of this coral species do not appear to represent an example of aposematic coloration.

Dietary specialists tend to be less susceptible to the effects of chemical defenses produced by their prey compared to generalist predators that feed upon a broader range of prey species. While many researchers have investigated the ability of insects to detoxify dietary allelochemicals, little research has been conducted in marine ecosystems. We investigated metabolic detoxification pathways in three species of butterflyfishes: the hard coral specialist feeder, Chaetodon multicinctus, and two generalist feeders, Chaetodon auriga and Chaetodon kleinii. Each species was fed tissue homogenate of the hard coral Porites lobata or the feeding deterrent compound homarine (found in the coral extract), and the expression and catalytic activity of cytochrome P450 (CYP) 3A-like and CYP2-like enzymes were examined after one-week of treatment. The P. lobata homogenate significantly induced content and catalytic activity of CYP2-like and CYP3A-like forms, by 2-3 fold and by 3-9 fold, respectively, in C. multicinctus. Homarine caused a significant decrease of CYP2-like and CYP3A-like proteins at the high dose in C. kleinii and 60-80% mortality in that species. Homarine also induced CYP3A-like content by 3-fold and catalytic activity by 2-fold in C. auriga, while causing non-monotonic increases in CYP2-like and CYP3A-like catalytic activity in C. multicinctus. Our results indicate that dietary exposure to coral homogenates and the feeding deterrent constituent within these homogenates caused species-specific modulation of detoxification enzymes consistent with the prey selection strategies of generalist and specialist butterflyfishes.

The transition between shallow and mesophotic coral reef communities in the tropics is characterized by a significant gradient in abiotic and biotic conditions that could result in potential trade-offs in energy allocation. The mesophotic reefs in the Bahamas and the Cayman Islands have a rich sponge fauna with significantly greater percent cover of sponges than in their respective shallow reef communities, but relatively low numbers of spongivores. Plakortis angulospiculatus, a common sponge species that spans the depth gradient from shallow to mesophotic reefs in the Caribbean, regenerates faster following predation and invests more energy in protein synthesis at mesophotic depths compared to shallow reef conspecifics. However, since P. angulospiculatus from mesophotic reefs typically contain lower concentrations of chemical feeding deterrents, they are not able to defend new tissue from predation as efficiently as conspecifics from shallow reefs. Nonetheless, following exposure to predators on shallow reefs, transplanted P. angulospiculatus from mesophotic depths developed chemical deterrence to predatory fishes. A survey of bioactive extracts indicated that a specific defensive metabolite, plakortide F, varied in concentration with depth, producing altered deterrence between shallow and mesophotic reef P. angulospiculatus. Different selective pressures in shallow and mesophotic habitats have resulted in phenotypic plasticity within this sponge species that is manifested in variable chemical defense and tissue regeneration at wound sites.

Over the past few decades, Caribbean coral reefs have undergone a phase shift from coral-dominated to algal-dominated communities due to several factors, including increased input of anthropogenic nutrients. With the decline in coral cover, sponges have also become more dom- inant members of Caribbean coral reef communities. Increased algal and sponge dominance on Caribbean reefs has led to an increase in the frequency of interaction between these 2 groups. This study used a factorial design to assess the independent and interactive effects of contact and elevated nutrient levels on 2 common members of these communities, the sponge Aplysina cauli- formis, and the macroalga Microdictyon marinum. Algal contact had a significant negative phys- iological effect on A. cauliformis, affecting both the host sponge and its cyanobacterial symbionts. While elevated nutrient levels had some positive effects on the sponge photosymbionts, this only occurred in the absence of algal contact or a shading/abrasion control, and elevated nutrient lev- els had a negative effect on the sponge holobiont. In contrast, M. marinum responded positively to experimentally enhanced nutrient levels and to sponge contact under ambient nutrient regimes, but was not affected by sponge contact under elevated nutrient concentrations. Stable isotope enrichment experiments showed that the alga's positive response to sponge contact was associated with nitrogen transfer from the sponge over the course of the experiment. Thus, while A. cauliformis facilitates increased productivity in M. marinum, algal contact competitively inhibits sponge condition.

Chemical investigation of the tunicate Trididemnum solidum resulted in the isolation of two new chlorinated compounds belonging to the didemnin class, along with two known compounds didemnin A and didemnin B. The structural determination of the compounds was based on extensive NMR and mass spectroscopic analysis. The isolated compounds 1–4 were tested for their anti-inflammatory activity using in vitro assays for inhibition of inducible nitric oxide synthase (iNOS) and nuclear factor-kappa B (NF-κB) activity. The anti-cell proliferative activity of the above compounds was also evaluated against four solid tumor cell lines.

Reports of marine sponge diseases have increased in recent years, but few etiologic agents have been identified. Aplysina red band syndrome (ARBS), a condition observed in the Caribbean sponge Aplysina cauliformis, is characterized by a rust-colored leading margin. Culture-independent methods (terminal restriction fragment length polymorphism and clone library analyses) were used to assess bacterial communities associated with healthy and ARBS-affected sponges from two locations over 2 years. Although the bacterial communities associated with healthy and ARBS-affected sponges were significantly different, the sponges maintained a core bacterial community across space, time, and health status. Ten terminal restriction fragments were shown to change significantly between sponge health conditions, with six increasing in abundance with disease and four decreasing. The prevalence of the photosymbiont Synechococcus spongiarum decreased with ARBS infection, suggesting a functional consequence of disease. After cultivating a red-pigmented Leptolyngbya strain from ARBS lesions, transmission studies were conducted to determine whether this organism was the ARBS pathogen. Despite significantly increased abundance of Leptolyngbya spp. in diseased sponges, signs of ARBS were not observed in healthy sponges following 24 days of contact with the cultured strain. Additional work with this model system is needed to increase our understanding of the dynamics of marine diseases.

Alcyonacean soft corals form major components of the biomass and biodiversity on many shallow Indo-Pacific reefs. In spite of the observed increase in marine diseases worldwide, disease has rarely been reported from this taxonomic group. Here, we describe a chronic tissue loss disease affecting soft corals of the genus Sinularia on reefs in Guam. The disease presents as a diffuse wrinkling of the otherwise smooth fingers, followed by tissue sloughing, necrosis, and disintegration. Until a cause has been confirmed, we propose the name Sinularia Tissue Loss Disease. This disease was first observed at low prevalence (

Marine caves are understudied ecosystems that are frequently associated with coral reef communities; many are tidally influenced and may host a highly diverse sponge fauna. Although each cave represents a distinct habitat likely structured by site-specific hydrographic processes, a more complete understanding of the ecology of these communities requires com parative studies. Based on a gradient of sponge cover within 5 Bahamian caves, we conducted a natural experiment in sponge-derived nutrient enrichment of nearby patch reefs. We tested the hypothesis that water exiting the cave during low tide provides a nutrient-rich resource that facilitates the diversity and health of nearby reef communities. The percent cover and diversity of corals surrounding the openings of caves were significantly higher than in similar habitats farther removed from these communities. There was a significant correlation between percent sponge cover within the caves and nitrate concentrations in seawater flowing out of the caves, and δ15N stable isotope signatures indicated enrichment of the nearby reefs by sponge-derived nitrate. Zooxanthellae abundance and total protein concentration were higher in corals from reefs near cave entrances, suggesting that those corals benefited more from cave nutrients than did corals farther from cave openings. In addition to corals, percent algal cover increased near cave entrances, but these potential competitors of corals were kept in check by increased levels of herbivory relative to sites removed from cave mouths. As global environmental changes continue to impact coral reef ecosystems, diversity ‘hot spots’, such as these marine caves, could serve as refuges and ‘seedbanks’ for nearby dwindling reef habitats.

Understanding prey response to predators and their utilization of sensory cues to assess local predation risk is crucial in determining how predator avoidance strategies affect population demographics. This study examined the antipredator behaviors of two ecologically similar species of Caribbean coral reef fish, Coryphopterus glaucofraenum and Gnatholepis thompsoni, and characterized their responses to different reef predators. In laboratory assays, the two species of gobies were exposed to predator visual cues (native Nassau grouper predator vs. invasive lionfish predator), damage-released chemical cues from gobies, and combinations of these, along with appropriate controls. Behavioral responses indicate that the two prey species differ in their utilization of visual and chemical cues. Visual cues from predators were decisive for both species’ responses, demonstrating their relative importance in the sensory hierarchy, whereas damage-released cues were a source of information only for C. glaucofraenum. Both prey species could distinguish between native and invasive predators and subsequently altered their antipredator responses.

Many butterflyfish species exhibit feeding preferences for scleractinian corals that represent a continuum from facultative to obligate strategies, and which affect their predicted responses to temporal and spatial changes in coral community structure. Less frequently studied are interactions between butterflyfishes and soft corals, even though the latter represent significant habitat and energetic resources on Indo-Pacific coral reefs. Changes to the benthic community structure of three disjunct back-reef communities on the leeward side of Guam were monitored over two decades, from 1994 to 2014. These communities include a reef that has remained stable and relatively pristine through time, a reef that is significantly impaired but has remained stable over time, and a reef that has experienced significant changes in response to anthropogenic and natural disturbances during this period. Density and behavior of six species of butterflyfishes, including an obligate hard corallivore, two soft corallivores, and three generalists of varied dietary breadth, were recorded at each of the sites in 1994, 2004, and 2014. The populations of butterflyfishes varied through time and space, and there were significant changes in foraging behaviors, largely associated with changes in the soft coral community. Moreover, the soft corallivores exhibited differential prey preferences, some of which were directly attributable to their ability to handle soft coral chemical defenses, while others appeared to be more opportunistic. Given this tight coupling between butterflyfishes and soft corals, this resource needs to be a focus of future studies.

Chemical diversity represents a measure of selective pressures acting on genotypic variability. In order to understand patterns of chemical ecology and biodiversity in the environment, it is necessary to enhance our knowledge of chemical diversity within and among species. Many sponges produce variable levels of secondary metabolites in response to diverse biotic and abiotic environmental factors. This study evaluated intra-specific variability in secondary metabolites in the common Indo-Pacific sponge Stylissa massa over various geographic scales, from local to ocean basin. Several major metabolites were quantified in extracts from sponges collected in American Samoa, Pohnpei, Saipan, and at several sites and depths in Guam. Concentrations of several of these metabolites varied geographically across the Pacific basin, with American Samoa and Pohnpei exhibiting the greatest differences, and Guam and Saipan more similar to each other. There were also significant differences in concentrations among different sites and depths within Guam. The crude extract of S. massa exhibited feeding deterrence against the omnivorous pufferfish Canthigaster solandri at natural concentrations, however, none of the isolated compounds was deterrent at the maximum natural concentrations observed, nor were mixtures of these compounds, thus emphasizing the need for bioassay-guided isolation to characterize specific chemical defenses. Antibacterial activity against a panel of ecologically relevant pathogens was minimal. Depth transplants, predator exclusion, and UV protection experiments were performed, but although temporal variability in compound concentrations was observed, there was no evidence that secondary metabolite concentration in S. massa was induced by any of these factors. Although the reasons behind the variability observed in the chemical constituents of S. massa are still in question, all sponges are not created equal from a chemical standpoint, and these studies provide further insights into patterns of chemical diversity within S. massa.

Diseases of marine organisms, and sponges in particular, are increasingly reported worldwide. Prior research indicates that the survival of sponges on reefs is due largely to their production of biologically active secondary metabolites that provide protection from a diversity of stressors. Aplysina Red Band Syndrome (ARBS) is an emerging disease affecting Caribbean rope sponges (Aplysina spp.), but it is not known whether secondary metabolites play a role in disease susceptibility and resistance. To investigate whether differences in secondary metabolites may explain variability in susceptibility to ARBS in Aplysina cauliformis, we used high performance liquid chromatography (HPLC) to generate chemical profiles from healthy tissue in both healthy and diseased sponges, and quantified peak areas for 15 metabolites. Analyses of healthy and diseased sponges revealed qualitative and quantitative differences in their chemical profiles. Aplysamine-1 and fistularin-3 were produced in significantly higher concentrations by healthy sponges, whereas aerothionin and 11-oxoaerothionin were found only in diseased sponges. At natural concentrations, extracts from both healthy and diseased sponges deterred feeding by an omnivorous reef fish. Fistularin-3 deterred feeding at concentrations found in healthy sponges, but not at concentrations found in diseased sponges. Aerothionin deterred feeding at concentrations found in diseased sponges, and may at least partially replace the loss of fistularin-3 as a feeding deterrent compound following pathogenesis, suggesting a trade-off in the production of feeding deterrent compounds. Extracts from healthy and diseased sponges inhibited bacterial growth, and both aplysamine-1 and fistularin-3 displayed selective antibacterial activity. Despite differences in secondary metabolite production between healthy and diseased sponges, the stress associated with ARBS does not appear to compromise the ability of A. cauliformis to maintain defenses against some of its natural enemies.

Territorial defense by the herbivorous damselfishes, Stegastes nigricans and S. lividus, benefits the hermatypic corals growing inside their territories. Coral diversity was significantly higher inside damselfish territories at 2 sites in Moorea, French Polynesia, and at 1 site in Guam, Mariana Islands. In Guam, this pattern was stable for at least 10 yr. Certain coral species, including Pocillopora damicornis, were found only inside damselfish territories at these sites. All fishes, including coral-feeding butterflyfishes, approaching territories of Stegastes spp. were vigorously chased. Colonies of P. damicornis inside territories were not preyed upon by any potential intruders. When transplanted outside of territories, colonies of P. damicornis were preyed upon rapidly by several species of butterflyfishes. Feeding rates of individual butterflyfishes on transplanted P. dami- cornis colonies were initially very high and decreased significantly over 30 min as accessible coral tissue was removed. In contrast, transplanted Porites rus, a major component of the reef outside of territories in Guam, attracted no predators. This study suggests that territorial damselfishes provide an associational defense for certain coral species that live inside their territories, whereas these species settling outside territories experience heavy predation, resulting in suppressed growth and negligible survival. This demonstrates the potential importance of this type of positive indirect interaction between species in structuring coral reef communities, that protection from predation by coral-feeding fishes is a mechanism by which damselfishes can enhance coral diversity on reefs, and that this association can remain stable for many years.

Although hybridization is a relatively widespread phenomenon in marine ecosystems, the ecological importance of hybrids is poorly understood. As crosses between two distinct genotypes, hybrids might express fitness characteristics similar to either parent species, or they might produce unique phenotypic attributes that make them more or less fit than either parent species. We identified a potential hybrid zone among soft corals in a back-reef community on Guam, where the broadcast-spawning species Sinularia maxima and S. polydactyla co-occur. Morphological and chemical traits confirmed the intermediate and unique status of the putative hybrid. Laboratory cross-fertilization experiments using S. maxima and S. polydactyla gametes demonstrated that barriers to hybridization are absent and that the laboratory-reared hybrids developed specific characteristics identified in adult hybrid field populations. Changes in the populations of each parent species and the hybrid were monitored over a decade. While popula...

Diseases of marine organisms, and corals in particular, have been reported with increas- ing frequency over the past 3 decades. Although little is known about resistance to disease in corals, a potential mechanism of defense is the production of antimicrobial compounds that protect corals from microbial pathogens. We assessed the antibacterial activity of crude aqueous extracts from 3 common Hawaiian reef corals, Montipora capitata, Porites lobata, and Pocillopora meandrina, against 9 strains of bacteria. The bacteria selected included known coral pathogens, potential marine pathogens found in human waste, and bacteria previously identified from the surfaces of Hawaiian corals. Extracts from all 3 coral species exhibited high levels of antibacterial activity. Overall, 54.3% of assays resulted in bacterial growth inhibition. In addition, 9.9% of extracts stimulated the growth of certain bacteria. Although levels of overall antibacterial activity did not differ significantly among coral species (ranging from 40.7% of assays for P. meandrina to 63.0% of assays for M. capitata), the 3 species exhibited a high degree of selectivity as demonstrated by significant variability in their effects toward specific bacteria. Extracts also exhibited intraspecific variability, both within and between sites, and significant site-by-species interactions were observed against 6 of 9 bacterial strains. In M. capitata, healthy corals had significantly higher levels of antibacterial activity than those affected by Montipora White Syndrome, and affected tissues had significantly higher levels of antibacterial activity than unaffected tissues. Variability in antibacterial activity of Hawaiian corals may, in part, explain differential susceptibility to disease at the colony, species, or population level.

Cnidarians rely on the innate immune defenses based on self/non-self recognition, signaling and effector responses to kill pathogens and heal wounds. Like other invertebrates, the immune system of cnidarians can be classified into several functional components and many of these elements have now been described in various cnidarian model systems. These include recognition receptors, toll-like receptors and peptidoglycan binding proteins, prophenoloxidase and melanin synthesis for an impermeable melanin barrier, anti-microbial proteins and molecules, reactive oxygen-producing and scavenging systems and wound repair and cellular systems. This chapter will summarize the current state of knowledge of cnidarian immune pathways and mechanisms. We will summarize the available data, guiding the reader through the steps of initiating and executing an immune response: recognition, signaling, effector and repair mechanisms. We will also explore the current research of constitutive immunity observed in healthy organisms and elicited immune responses seen in naturally infected organisms or organisms exposed to live pathogens or pathogen associated molecular patterns. We will connect these known immune pathways with how they are expressed and regulated and how this may influence the wide variation in disease resistance observed in the field, both within and between species. The overarching goal of this chapter is to take the reader from a genomic to phenotypic perspective, while keeping pathways and mechanisms in a whole organism and ecological context, whenever possible.

Emerging diseases of marine invertebrates have been implicated as one of the major causes of the continuing decline in coral reefs worldwide. To date, most of the focus on marine diseases has been aimed at hard (scleractinian) corals, which are the main reef builders worldwide. However, soft (alcyonacean) corals are also essential components of tropical reefs, representing food, habitat and the 'glue' that consolidates reefs, and they are subject to the same stressors as hard corals. Sinularia maxima and S. polydactyla are the dominant soft corals on the shallow reefs of Guam, where they hybridize. In addition to both parent species, the hybrid soft coral population in Guam is particularly affected by Sinularia tissue loss disease. Using label-free shotgun proteomics, we identified differences in protein expression between healthy and diseased colonies of the hybrid S. maxima × S. polydactyla. This study provided qualitative and quantitative data on specific proteins that were differentially expressed under the stress of disease. In particular, metabolic proteins were down-regulated, whereas proteins related to stress and to symbiont photosynthesis were up-regulated in the diseased soft corals. These results indicate that soft corals are responding to pathogenesis at the level of the proteome, and that this label-free approach can be used to identify and quantify protein biomarkers of sub-lethal stress in studies of marine disease.

A substantial and increasing number of reports have documented dramatic changes and continuing declines in Caribbean coral reef communities over the past 2 decades. To date, the majority of disease reports have focused on scleractinian corals, whereas sponge diseases have been less frequently documented. In this study, we describe Aplysina red band syndrome (ARBS) affecting Caribbean rope sponges of the genus Aplysina observed on shallow reefs in the Bahamas. Visible signs of disease presence included 1 or more rust-colored leading edges, with or without a trailing area of necrotic tissue, such that the lesion forms a contiguous band around part or all of the sponge branch. Microscopic examination of the leading edge of the disease margin indicated that a cyanobacterium was consistently responsible for the coloration. Although the presence of this distinctive coloration was used to characterize the diseased state, it is not yet known whether this cyanobacterium is directly responsible for disease causation. The prevalence of ARBS declined significantly from July to October 2004 before increasing above July levels in January 2005. Transmission studies in the laboratory demonstrated that contact with the leading edge of an active lesion was sufficient to spread ARBS to a previously healthy sponge, suggesting that the etiologic agent, currently undescribed, is contagious. Studies to elucidate the etiologic agent of ARBS are ongoing. Sponges are an essential component of coral reef communities and emerging sponge diseases clearly have the potential to impact benthic community structure on coral reefs.

In clonal organisms, such as corals, one consequence of partial predation may be an elaboration of defenses in remaining portions of the clone, thereby reducing the probability or sever- ity of future predation events. Inducible defenses have been found in terrestrial and marine plants and in several taxa of marine invertebrates. Predators can detect differences in various aspects of prey quality that translate into preferences for certain prey items. Differences in quantity or types of defenses may determine which species, individuals or parts of a prey item are consumed. Coral- feeding butterflyfishes show distinct preferences for certain coral species, and may prefer particular individuals of a species over others. This study examines the potential for inducible defenses in a hard coral in response to grazing by a natural coral predator, the butterflyfish Chaetodon multicinctus. Pairs of genetically identical fragments of the Hawaiian coral Porites compressa were exposed to grazed and ungrazed treatments. These colonies were then offered to naive fish in preference tests at various intervals following the treatment period. Grazing by butterflyfishes induced changes in polyp behavior (prolonged withdrawal of coral polyps) in the short term, and increases in nematocyst density over the longer term, and these changes were associated with reductions in palatability and subsequent predation rates on the damaged corals. These inducible responses may play a role in regulating the intensity of grazing, and ultimately territory size and the density of corallivorous reef fishes.

Background Marine diseases are of increasing concern for coral reef ecosystems, but often their causes, dynamics and impacts are unknown. The current study investigated the epidemiology of Aplysina Red Band Syndrome (ARBS), a disease affecting the Caribbean sponge Aplysina cauliformis, at both the individual and population levels. The fates of marked healthy and ARBS-infected sponges were examined over the course of a year. Population-level impacts and transmission mechanisms of ARBS were investigated by monitoring two populations of A. cauliformis over a three year period using digital photography and diver-collected data, and analyzing these data with GIS techniques of spatial analysis. In this study, three commonly used spatial statistics (Ripley’s K, Getis-Ord General G, and Moran’s Index) were compared to each other and with direct measurements of individual interactions using join-counts, to determine the ideal method for investigating disease dynamics and transmission mechanisms in this system. During the study period, Hurricane Irene directly impacted these populations, providing an opportunity to assess potential storm effects on A. cauliformis and ARBS. Results Infection with ARBS caused increased loss of healthy sponge tissue over time and a higher likelihood of individual mortality. Hurricane Irene had a dramatic effect on A. cauliformis populations by greatly reducing sponge biomass on the reef, especially among diseased individuals. Spatial analysis showed that direct contact between A. cauliformis individuals was the likely transmission mechanism for ARBS within a population, evidenced by a significantly higher number of contact-joins between diseased sponges compared to random. Of the spatial statistics compared, the Moran’s Index best represented true connections between diseased sponges in the survey area. This study showed that spatial analysis can be a powerful tool for investigating disease dynamics and transmission in a coral reef ecosystem.

The application of proteomics to marine sciences has increased in recent years because the proteome represents the interface between genotypic and phenotypic variability and, thus, corresponds to the broadest possible biomarker for eco-physiological responses and adaptations. Likewise, proteomics can provide important functional information regarding biosynthetic pathways, as well as insights into mechanism of action, of novel marine natural products. The goal of this review is to (1) explore the application of proteomics methodologies to marine systems, (2) assess the technical approaches that have been used, and (3) evaluate the pros and cons of this proteomic research, with the intent of providing a critical analysis of its future roles in marine sciences. To date, proteomics techniques have been utilized to investigate marine microbe, plant, invertebrate, and vertebrate physiology, developmental biology, seafood safety, susceptibility to disease, and responses to environmental change. However, marine proteomics studies often suffer from poor experimental design, sample processing/optimization difficulties, and data analysis/interpretation issues. Moreover, a major limitation is the lack of available annotated genomes and proteomes for most marine organisms, including several "model species". Even with these challenges in mind, there is no doubt that marine proteomics is a rapidly expanding and powerful integrative molecular research tool from which our knowledge of the marine environment, and the natural products from this resource, will be significantly expanded.

Chemical investigations of two collections of the deep reef Caribbean sponge Plakortis angulospiculatus resulted in the isolation of a new compound (1) along with the known compound spiculoic acid B (2) belonging to the spiculoic acid class and four other new compounds (3-6) belonging to the zyggomphic acid class. Three new aromatic compounds (7-9) were isolated from the Caribbean sponge Plakortis halichondrioides. The structural determination of the compounds was based on extensive NMR and mass spectroscopic analysis. The isolated compounds 1-7 were tested for their anti-inflammatory activity using in vitro assays for inhibition of inducible nitric oxide synthase and nuclear factor kappa B (NFκB) activity, as well as inhibition of intracellular reactive oxygen species generation as a result of oxidative stress. The cytotoxicity of these compounds was also evaluated to determine the selectivity index of their bioactivity with respect to cytotoxicity. Compounds 1 and 4 were more potent than the positive control in inhibiting NFκB activity and had IC(50) values of 0.47 and 2.28 μM, respectively.

Scleractinian corals appear to be increasingly susceptible to pathogenic diseases, yet it is poorly understood why certain individuals, populations or species are more susceptible to diseases than others. Clearly an understanding of mechanisms of disease resistance in corals is essential to our understanding of patterns of disease incidence and virulence; this work must begin by examining the colony and population levels of organization. The Caribbean coral Siderastrea siderea exhibits variability in susceptibility to dark spot syndrome (DSS), a disease of unknown origin that can result in tissue necrosis. On the reef scale, variability in DSS prevalence in S. siderea occurred through time, but was not correlated with site, seawater temperature or depth. We monitored colonies of S. siderea affected by DSS, as well as their nearest neighbor controls, for 2 years in the Bahamas and found a marked decline in extent of DSS infection in October of both years. A preliminary survey of antimicrobial activity in S. siderea indicated selective activity against certain ecologically relevant bacteria. To assess whether changes in chemical defenses were responsible for the observed temporal variability in DSS prevalence, we sampled S. siderea for qualitative and quantitative analysis of chemical variability between resistant and susceptible colonies of S. siderea. These data suggest that phenotypic plasticity in antimicrobial activity may impact microbial settlement and/or survival.

The bioassay- and spectroscopic-guided fractionation of the antimalarial extract from a Jamaican sponge, Plakortis sp., resulted in the isolation of three metabolites. The previously reported bromoaromatic filiformin (1) was obtained from our sample of Plakortis sp., and the potential origins of this compound are discussed. The peroxide-containing metabolite, plakortide F (2), is a more typical Plakortis metabolite and was shown to exhibit significant activity against Plasmodium falciparum in vitro. The isolation, structure, and bioactivity of a new lactone, plakortone G (3), are also reported.

We compared species presence, abundance, and size characteristics of fish in three brackish, coastal marshes at Humacao, Roosevelt Roads, and Boqueron, Puerto Rico, in February and March 1988. The three marsh ecosystems were similar with respect to the presence of large expanses of open water bordered by emergent vegetation, creeks, and mangroves, and all had some recreational use. We sampled fish using gill nets. Tilapia (Oreochromis) mossambica were the most abundant fish, accounting for 55–79% of the samples at all three marshes. Overall, tilapia were both the largest (North Lagoon) and the smallest (Frontera Creek) at Humacao. Tilapia were most common in open lagoons rather than creeks or bays (except for Mandri Creek), and their distribution seemed unrelated to salinity. Tarpon (Megalops atlantica) were more abundant at low salinities, whereas other fish were more abundant at higher salinities.

We surveyed several offshore islands around Culebra for nesting Zenaida Doves and seabirds to determine if the doves preferred islands with nesting seabirds, and whether or not the doves selected the same types of sites on all islands. Doves nested on eight of the islands with seabirds, and did not nest on four islands that were devoid of nesting seabirds. Nest-site characteristics differed among the islands, although there were no inter-island differences with respect to nest size, slope at the nest, and percent of the nest visible from above and from 1 m. Thus slope and nest visibility seem to be significant habitat cues for nesting Zenaida Doves.