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Despite increasing research effort on coral diseases, little is known about factors driving disease dynamics on the Great Barrier Reef (GBR). This is the first study to investigate the temporal patterns of coral disease prevalence and potential drivers of disease around Heron Island, in the southern Capricorn Bunker sector of the GBR. Surveys were conducted in two austral summers and three winters between November 2007 and August 2009 on six sites around the island. Six diseases were detected: brown band syndrome (BrB), growth anomalies (GA), ulcerative white spots (UWS), white syndrome (WS), skeletal eroding band disease (SEB) and black band disease (BBD). The lowest overall mean disease prevalence was 1.87 ± 0.75% (mean ± SE) in November 2007 and the highest 4.22 ± 1.72% in August 2008. There was evidence of seasonality for two diseases: BrB and UWS. This is the first study to report a higher prevalence of BrB in the winter. BrB had a prevalence of 3.29 ± 0.58% in August 2008 and 1.53 ± 0.28% in August 2009, while UWS was the most common syndrome in the summer with a prevalence of 1.12 ± 0.31% in November 2007 and 2.67 ± 0.52% prevalence in January 2008. The prevalence of GAs and SEB did not depend on the season, although the prevalence of GAs increased throughout the study period. WS had a slightly higher prevalence in the summer, but its overall prevalence was low (

This study compared the effect of heat stress on coral-associated bacterial communities among juveniles of the coral, Acropora tenuis, hosting different Symbiodinium types. In comparison to a control temperature treatment (28 degrees C), we documented dramatic changes in bacterial associates on juvenile corals harbouring ITS 1 type D Symbiodinium when placed in a high (32 degrees C) temperature treatment. In particular, there was a marked increase in the number of retrieved Vibrio affiliated sequences, which coincided with a 44% decline in the photochemical efficiency of the D-juveniles. Interestingly, these Vibrio sequences affiliated most closely with the coral pathogen, Vibrio coralliilyticus, which has been implicated in some coral disease outbreaks. In contrast, A. tenuis hosting ITS 1 type C1 Symbiodinium did not exhibit major bacterial shifts in the elevated temperature treatment, indicating a more stable bacterial community during thermal stress; concomitantly a decline (10%) in photochemical efficiency was minimal for this group. D juveniles that had been exposed to moderately elevated sea temperatures (30 degrees C) in the field before being placed in the control temperature treatment displayed a decrease in the number of Vibrio affiliated sequences and bacterial profiles shifted to become more similar to profiles of corals harbouring type C1 Symbiodinium. In combination, these results demonstrate that thermal stress can result in shifts in coral-associated bacterial communities, which may lead to deteriorating coral health. The lower resilience of A. tenuis to thermal stress when harbouring Symbiodinium D highlights the importance of inter-kingdom interactions among the coral host, dinoflagellate endosymbiont and bacterial associates for coral health and resilience.

Dimethylsulfoniopropionate (DMSP) and dimethylsulfide (DMS) are key compounds in the global sulfur cycle. Moreover, DMS is particularly important in climate regulation owing to its role in cloud formation. Reef building corals are major contributors to the production of these two compounds and also form diverse and complex associations with bacteria, which are known to play a crucial role in the degradation of DMSP and DMS. Here, we highlight an extensive overlap between bacterial species implicated in DMSP/DMS degradation and those associated with corals, leading to the hypothesis that these two compounds play a major role in structuring coral-associated bacterial communities, with important consequences for coral health and the resilience of coral reefs. We also explore the publically available metagenome databases and show that genes implicated in DMSP metabolism are abundant in the viral component of coral-reef-derived metagenomes, indicating that viruses can act as a reservoir for such genes.

Immunity is a key life history trait that may explain hierarchies in the susceptibility of corals to disease and thermal bleaching, two of the greatest current threats to coral health and the persistence of tropical reefs. Despite their ongoing and rapid global decline, there have been few investigations into the immunity mechanisms of reef-building corals. Variables commonly associated with invertebrate immunity, including the presence of melanin, size of melanin-containing granular cells, and phenoloxidase (PO) activity, as well as concentrations of fluorescent proteins (FPs), were investigated in hard (Scleractinia) and soft (Alcyonacea) corals spanning 10 families from the Great Barrier Reef. Detectable levels of these indicators were present in all corals investigated, although relative investment differed among coral taxa. Overall levels of investment were inversely correlated to thermal bleaching and disease susceptibility. In addition, PO activity, melanin-containing granular cell size, and FP concentration were each found to be significant predictors of susceptibility and thus may play key roles in coral immunity. Correlative evidence that taxonomic (family-level) variation in the levels of these constituent immunity parameters underpins susceptibility to both thermal bleaching and disease indicates that baseline immunity underlies the vulnerability of corals to these two threats. This reinforces the necessity of a holistic approach to understanding bleaching and disease in order to accurately determine the resilience of coral reefs.

A phenotypic and phylogenetic comparison of geographically disparate isolates of the coral pathogen Vibrio coralliilyticus was conducted to determine whether the bacterium exists as a single cosmopolitan clonal population, which might indicate rapid spread of a pandemic strain, or is grouped into endemic and genotypically distinct strains. All strains included in this study displayed similar phenotypic characteristics to those of the typed V. coralliilyticus strain LMG 20984T. Five phylogenetic marker genes (16S, rpoA, recA, pyrH and dnaJ) frequently used for discriminating closely related Vibrio species and a zinc-metalloprotease gene (vcpA) linked to pathogenicity were sequenced in 13 V. coralliilyticus isolates collected from corals, bivalves, and their surrounding seawater in the Red and Caribbean Seas, and Indian, Pacific and Atlantic Oceans. A high level of genetic polymorphism was observed with all isolates possessing unique genotypes at all six genetic loci examined. No consistent lineage structure was observed within the marker genes and homologous recombination was detected in the 16S and vcpA genes, suggesting that V. coralliilyticus does not possess a highly clonal population structure. Interestingly, two geographically distinct (Caribbean/south-Atlantic and Indo-Pacific/north-Atlantic) and highly divergent clades were detected within the zinc-metalloprotease gene, but it is not known if these clades correspond to phenotypic differences in virulence. These findings stress the need for a multi-locus approach for inferring V. coralliilyticus phylogeny and indicate that populations of this bacterium are likely an endemic component of coral reef ecosystems globally.

A limited diversity of character states for reproductive traits and a robust phylogeny make scleractinian corals an ideal model organism with which to explore the evolution of life-history traits. Here, we explore systematic and biogeographical patterns in the reproductive biology of the Scleractinia within the context of a new molecular phylogeny and using reproductive traits from nearly 400 species. Our analyses confirm that coral sexuality is highly conserved, and mode of larval development is relatively plastic. An overabundance of species with autotrophic larvae in the eastern Pacific and Atlantic is most likely the result of increased capacity for long-distance dispersal conferred by vertical transmission of symbiotic zooxanthellae. Spawning records from diverse biogeographical regions indicate that multispecies spawning occurs in all speciose coral assemblages. A new quantitative index of spawning synchrony shows peaks at mid-tropical latitudes in the Indo-Pacific, influenced in part by two spawning seasons in many species on equatorial reefs.

Reef-building corals are threatened by increasing sea surface temperatures resulting from global climate change. Whether corals can adapt to increasing temperatures over the course of generations will depend in part on heritable variation in thermal physiology and dispersal potential, which may serve as the raw material for natural selection. To investigate whether such variation exists in coral populations, and build a framework for identifying the coral-specific genetic factors involved, we performed controlled crosses between 3 genetically distinct colonies of the branching coral Acropora millepora. We compared the families of larvae (which in this species naturally lack symbionts) for several physiological traits, and observed between-family differences in nearly every case. Using larvae cultured at standard and elevated temperatures, we measured the developmental decrease in protein content and the expression of candidate heat response genes. We used an in vivo assay for mitochondrial enzyme activity to evaluate the metabolic response to temperature changes in individual larvae. We also compared the responsiveness of larvae from different families to a nat- ural settlement cue to gain insights into long-range dispersal potential. Partitioning the components of total phenotypic variance confirmed the existence of additive genetic effects for settlement rates and βγ-crystallin expression, while variance in mitochondrial Q10 and the expression of actin and Hsp16 were driven by non-additive effects. The phenotypic variance observed among the small num- ber of families analyzed here suggests the existence of considerable heritable variation in natural coral populations, which supports the possibility of effective adaptive responses to climate change.

Black band disease (BBD) consists of a mat-forming microbial consortium that migrates across coral colonies causing rapid tissue loss. Although BBD-associated microbial communities have been well characterized, little is known regarding how these complex bacterial consortia develop. This study analyzed successional changes in microbial communities leading to the development of BBD. Long-term monitoring of tagged corals throughout outbreaks of BBD in the central Great Barrier Reef documented cyanobacterium-infected lesions, herein termed cyanobacterial patch(es) (CP), which were macroscopically distinct from BBD and preceded the onset of BBD in 19% of the cases. Dominant cyanobacteria within CP lesions were morphologically distinct from ones dominating BBD lesions. Clone libraries and terminal restriction fragment length polymorphism analysis confirmed shifts within cyanobacterial assemblages, from Blennothrix sp.-affiliated sequences dominating CP lesions, to Oscillatoria sp.-affiliated sequences, similar to those retrieved from other BBD samples worldwide, dominating BBD lesions. Bacterial 16S ribosomal RNA clone libraries also showed shifts in bacterial ribotypes during transitions from CP to BBD, with Alphaproteobacteria-affiliated sequences dominant in CP libraries, whereas gammaproteobacterial and cyanobacterial ribotypes were more abundant in BBD clone libraries. Sequences affiliated with organisms identified in sulfur cycling were commonly retrieved from lesions showing characteristic field signs of BBD. As high sulfide concentrations have been implicated in BBD-mediated coral tissue degradation, proliferation of a microbial community actively involved in sulfur cycling potentially contributes to the higher progression rates found for BBD compared with CP lesions. Results show how microbial colonization of indistinct lesions may facilitate a common coral disease with proven ecological effects on coral populations.

The coral holobiont consists of the host and its microbial partners, including the dinoflagellate endosymbiont Symbiodinium and bacteria living both on and within coral tissues. Although genetically different, Symbiodinium types have been shown to differentially affect the physiology of the coral host; their effects on the bacterial partners in the association are unknown. The present study compares profiles of the bacterial communities associated with juvenile corals of Acropora millepora and A. tenuis that had been experimentally infected with 2 different clades of Symbiodinium, Clade C1 and D, to investigate possible interactions between bacterial and Symbiodinium communities. Three culture-independent 16S rRNA gene profiling methods (clone library construction, terminal restriction length polymorphism and denaturing gradient gel electrophoresis) revealed no discernible pattern in bacterial communities on 9 mo old juvenile corals containing different clades of zooxanthellae, suggesting that coral-associated bacteria are not linked to Symbiodinium types in hospite in early ontogeny. In contrast to bacterial profiles of adult corals, bacterial communities associated with juvenile corals were highly variable, indicating that bacterial associates are not conserved in these early stages. When 12 mo old juveniles were sampled again in summer, bacterial communities associated with A. tenuis hosting Clade D Symbiodinium were dominated by sequences affiliating with Vibrio species, indicating that corals harbouring this symbiont may be more susceptible to temperature stress, allowing growth of opportunistic microbial community members possibly detrimental to coral health.

The majority of reef-building corals acquire their obligate algal symbionts (Symbiodinium) from the environment. However, factors shaping the initial establishment of coral-algal symbioses, including parental effects, local environmental conditions and local availability of symbionts, are not well understood. This study monitored the uptake and maintenance of Symbiodinium in juveniles of two common corals, Acropora tenuis and Acropora millepora, that were reciprocally explanted between sites where adult colonies host different types of Symbiodinium. We found that coral juveniles were rapidly dominated by type D Symbiodinium, even though this type is not found in adult colonies (including the parental colonies) in four out of the five study populations. Furthermore, type D Symbiodinium was found in less than one-third of a wide range of coral species (n > 50) sampled at the two main study sites, suggesting that its dominance in the acroporid juveniles is not because it is the most abundant local endosymbiotic type. Moreover, dominance by type D was observed irrespective of the light intensity to which juveniles were exposed in a field study. In summary, despite its relatively low abundance in coral assemblages at the study sites and irrespective of the surrounding light environment, type D Symbiodinium is the main symbiont type initially acquired by juveniles of A. millepora and A. tenuis. We conclude that during early ontogeny in these corals, there are few barriers to the uptake of Symbiodinium types which differ from those found in parental colonies, resulting in dominance by a highly infectious and potentially opportunistic symbiont.

Marine bacteria play a central role in the degradation of dimethylsulfoniopropionate (DMSP) to dimethyl sulfide (DMS) and acrylic acid, DMS being critical to cloud formation and thereby cooling effects on the climate. High concentrations of DMSP and DMS have been reported in scleractinian coral tissues although, to date, there have been no investigations into the influence of these organic sulfur compounds on coral-associated bacteria. Two coral species, Montipora aequituberculata and Acropora millepora , were sampled and their bacterial communities were characterized by both culture-dependent and molecular techniques. Four genera, Roseobacter, Spongiobacter, Vibrio , and Alteromonas , which were isolated on media with either DMSP or DMS as the sole carbon source, comprised the majority of clones retrieved from coral mucus and tissue 16S rRNA gene clone libraries. Clones affiliated with Roseobacter sp. constituted 28% of the M. aequituberculata tissue libraries, while 59% of the clones from the A. millepora libraries were affiliated with sequences related to the Spongiobacter genus. Vibrio spp. were commonly isolated from DMS and acrylic acid enrichments and were also present in 16S rRNA gene libraries from coral mucus, suggesting that under “normal” environmental conditions, they are a natural component of coral-associated communities. Genes homologous to dddD , and dddL , previously implicated in DMSP degradation, were also characterized from isolated strains, confirming that bacteria associated with corals have the potential to metabolize this sulfur compound when present in coral tissues. Our results demonstrate that DMSP, DMS, and acrylic acid potentially act as nutrient sources for coral-associated bacteria and that these sulfur compounds are likely to play a role in structuring bacterial communities in corals, with important consequences for the health of both corals and coral reef ecosystems.

Recurring summer outbreaks of black band disease (BBD) on an inshore reef in the central Great Barrier Reef (GBR) constitute the first recorded BBD epizootic in the region. In a 2.7 year study of 485 colonies of Montipora species, BBD affected up to 10 per cent of colonies in the assemblage. Mean maximum abundance of BBD reached 16±6 colonies per 100 m 2 ( n =3 quadrats, each 100 m 2 ) in summer, and decreased to 0–1 colony per 100 m 2 in winter. On average, BBD lesions caused 40 per cent tissue loss and 5 per cent of infections led to whole colony mortality. BBD reappearance on previously infected colonies and continuous tissue loss after the BBD signs had disappeared suggest that the disease impacts are of longer duration than indicated by the presence of characteristic signs. Rates of new infections and linear progression of lesions were both positively correlated with seasonal fluctuations in sea water temperatures and light, suggesting that seasonal increases in these environmental parameters promote virulence of the disease. Overall, the impacts of BBD are greater than previously reported on the GBR and likely to escalate with ocean warming.

Patterns in the diversity of bacterial communities associated with three species of Acropora (Acropora millepora, Acropora tenuis and Acropora valida) were compared at two locations (Magnetic Island and Orpheus Island) on the Great Barrier Reef to better understand the nature and specificity of coral-microbial symbioses. Three culture-independent techniques demonstrated consistent bacterial communities among replicate samples of each coral species, confirming that corals associate with specific microbiota. Profiles were also conserved among all three species of Acropora within each location, suggesting that closely related corals of the same genus harbor similar bacterial types. Bacterial community profiles of A. millepora at Orpheus Island were consistent in samples collected throughout the year, indicating a stable community despite temporal changes. However, DGGE and T-RFLP profiles differed on corals from different reefs. Nonmetric multidimensional scaling of T-RFLP profiles showed that samples grouped according to location rather than coral species. Although similar sequences were retrieved from clone libraries of corals at both Magnetic and Orpheus Island, differences in the relative dominant bacterial ribotypes within the libraries drive bacterial community structure at different geographical locations. These results indicate certain bacterial groups associated specifically with corals, but the dominant bacterial genera differ between geographically-spaced corals.

The extent of larval dispersal on coral reefs has important implications for the persistence of coral reef metapopulations, their resilience and recovery from an increasing array of threats, and the success of protective measures. This article highlights a recent dramatic increase in research effort and a growing diversity of approaches to the study of larval retention within (self-recruitment) and dispersal among (connectivity) isolated coral reef populations. Historically, researchers were motivated by alternative hypotheses concerning the processes limiting populations and structuring coral reef assemblages, whereas the recent impetus has come largely from the need to incorporate dispersal information into the design of no-take marine protected area (MPA) networks. Although the majority of studies continue to rely on population genetic approaches to make inferences about dispersal, a wide range of techniques are now being employed, from small-scale larval tagging and paternity analyses, to large-scale biophysical circulation models. Multiple approaches are increasingly being applied to cross-validate and provide more realistic estimates of larval dispersal. The vast majority of empirical studies have focused on corals and fishes, where evidence for both extremely local scale patterns of self-recruitment and ecologically significant connectivity among reefs at scales of tens of kilometers (and in some cases hundreds of kilometers) is accumulating. Levels of larval retention and the spatial extent of connectivity in both corals and fishes appear to be largely independent of larval duration or reef size, but may be strongly influenced by geographic setting. It is argued that high levels of both self-recruitment and larval import can contribute to the resilience of reef populations and MPA networks, but these benefits will erode in degrading reef environments.

The resilience of Symbiodinium harboured by corals is dependent on the genetic diversity and extent of connectivity among reef populations. This study presents genetic analyses of Great Barrier Reef (GBR) populations of clade C Symbiodinium hosted by the alcyonacean coral, Sinularia flexibilis. Allelic variation at four newly developed microsatellite loci demonstrated that Symbiodinium populations are genetically differentiated at all spatial scales from 16 to 1,360 km (pairwise ΦST = 0.01–0.47, mean = 0.22); the only exception being two neighbouring populations in the Cairns region separated by 17 km. This indicates that gene flow is restricted for Symbiodinium C hosted by S. flexibilis on the GBR. Patterns of population structure reflect longshore circulation patterns and limited cross-shelf mixing, suggesting that passive transport by currents is the primary mechanism of dispersal in Symbiodinium types that are acquired horizontally. There was no correlation between the genetic structure of Symbiodinium populations and their host S. flexibilis, most likely because different factors affect the dispersal and recruitment of each partner in the symbiosis. The genetic diversity of these Symbiodinium reef populations is on average 1.5 times lower on inshore reefs than on offshore reefs. Lower inshore diversity may reflect the impact of recent bleaching events on Sinularia assemblages, which have been more widespread and severe on inshore reefs, but may also have been shaped by historical sea level fluctuations or recent migration patterns.

Previously undetected earlier phases of the coral disease 'atramentous necrosis' are documented and described. New observations relating to the occurrence of initial stages and progression of the disease are reported, and potential cause(s) are examined. In direct contrast to earlier published findings, temperature data indicated that occurrence of early bleached stages of atramentous necrosis is not correlated with warmer water temperatures; however, the relationship between temperature and disease prevalence is still unclear.

The abundance and distribution of free-living dinoflagellates in the genus Symbiodinium have important implications for the ecology of coral reefs, determining both the symbionts available to newly recruited corals and symbiont types available for uptake by adult corals during environmental stress. However, little is known about where symbiotic dinoflagellates reside outside the host, due to the difficulty of capturing and detecting unicellular organisms in the marine environment. This study presents a successful protocol for sampling Symbiodinium from both the benthos and the water column. Comparisons of two detection methods for enumerating Symbiodinium indicated that conventional microscope analysis is accurate and more efficient when estimating Symbiodinium densities in sediment samples, while an automated particle counter (FlowCAM) was more efficient in detecting cells in the water column where densities are low. Symbiodinium densities were found to be relatively high (1000–4000 cells/mL) in sediment samples and much lower (up to 80 cells/mL) in the water column, indicating that the free-living form resides mainly in the benthos. Symbiodinium densities were found to be highly variable spatially, differing significantly between two reef locations. Within sites, elevated densities of Symbiodinium along reef margins combined with significant decreases in densities one meter away from the reef, suggest that cells aggregate within the reef habitat.

A balanced approach to coral disease investigation is critical for understanding the global decline of corals. Such an approach should involve the proper use of biomedical concepts, tools, and terminology to address confusion and promote clarity in the coral disease literature. Investigating disease in corals should follow a logical series of steps including identification of disease, systematic morphologic descriptions of lesions at the gross and cellular levels, measurement of health indices, and experiments to understand disease pathogenesis and the complex interactions between host, pathogen, and the environment. This model for disease investigation is widely accepted in the medical, veterinary and invertebrate pathology disciplines. We present standard biomedical rationale behind the detection, description, and naming of diseases and offer examples of the application of Koch's postulates to elucidate the etiology of some infectious diseases. Basic epidemiologic concepts are introduced to help investigators think systematically about the cause(s) of complex diseases. A major goal of disease investigation in corals and other organisms is to gather data that will enable the establishment of standardized case definitions to distinguish among diseases. Concepts and facts amassed from empirical studies over the centuries by medical and veterinary pathologists have standardized disease investigation and are invaluable to coral researchers because of the robust comparisons they enable; examples of these are given throughout this paper. Arguments over whether coral diseases are caused by primary versus opportunistic pathogens reflect the lack of data available to prove or refute such hypotheses and emphasize the need for coral disease investigations that focus on: characterizing the normal microbiota and physiology of the healthy host; defining ecological interactions within the microbial community associated with the host; and investigating host immunity, host-agent interactions, pathology, pathogenesis, and factors that promote the pathogenicity of the causative agent(s) of disease.

Recovery of degraded reefs is dependent on the settlement of coral larvae into habitats typically dominated by benthic algae, so that benthic algae may play pivotal roles in coral settlement and reef recovery. Here we demonstrate that waterborne influences of macroalgae could affect coral settlement before larvae contact reef substrata and that such effects vary between macroalgae. We tested for waterborne effects of algae on both pre-settlement behaviour and settlement of larvae of the coral Acropora millepora onto live fragments of the crustose coralline alga Hydrolithon reinboldii. Treatments comprised seawater collected from aquaria that had previously contained 1 of 3 macroalgae common on degraded reefs. The foliose brown macroalga, Lobophora variegata, enhanced coral settlement by 40% relative to substratum control treatments. In contrast, the filamentous green macroalga Chlorodesmis fastigiata ('turtle weed'), hindered coral settlement by delaying settlement of larvae, although final settlement was similar to that in control treatments. Padina sp., a foliose brown macroalga closely related to L. variegata, reduced coral settlement by 30% compared with substratum controls. The demonstration of waterborne effects suggests that macroalgae can influence coral settlement before larvae reach reef substrata, even on a crustose coralline alga known to induce settlement, and even where the immediate settlement location is free of macroalgal cover. These results demonstrate the complexity in the mechanisms underlying the effects that over-abundant macroalgal growth may have on reef recovery. These effects have critical implications for the ecological resilience of coral reefs, especially as climate change increases the frequency and severity of disturbances to reefs.

Skeletal eroding band (SEB), which manifests as dense aggregations of the ciliate Halofolliculina corallasia, was the first coral disease described from the Indo-Pacific. Little is known about its etiology or impact. This study describes the distribution, prevalence and host range of SEB on a 500 km extent of the Great Barrier Reef (GBR), together with in situ rates of progression and infection following experimental injury. SEB occurred on 90–100% of reefs surveyed (n=18) in each of 3 years, demonstrating that SEB is widely distributed and persistent. SEB had the highest prevalence of any disease, affecting approximately 2% of 283,486 scleractinians and hydrocorals surveyed. Its host range was large, affecting 12 families and at least 82 scleractinian species, as well as the hydrocoral, Millepora. Corals in the families Pocilloporidae and Acroporidae were most susceptible, the former being up to five times more susceptible than other families. Progressive tissue loss was recorded on 95% of Acropora muricata colonies monitored (n = 18), with rates of SEB progression averaging ∼2 mm/day. Injury experiments demonstrated that H. corallasia, the putative pathogen of SEB, readily colonised recently exposed coral skeleton in the absence of a vector, but did not colonise intact coral tissue. Invading ciliates failed to form band-like aggregations associated with progressive tissue loss on any of three coral species tested experimentally, suggesting that, while H. corallasia readily colonises recently exposed coral skeleton, it may not be sufficient in itself to cause tissue mortality. Interactions with additional agents or factors, increasing ciliate virulence or lowering disease resistance of coral hosts may be required before halofolliculinid infections become associated with tissue loss.

The photosystem II (PSII) herbicide diuron is commonly used within catchments that drain into the Great Barrier Reef lagoon, where it potentially reduces photosynthesis in Symbiodinium, the dinoflagellate symbiont associated with reef corals. Here we examine the importance of energy (carbohydrates) derived from photosynthesis to the gametogenesis of corals following long-term, experimental exposures to diuron. Two broadcast spawning corals, Acropora tenuis and A. valida, and a brooding coral, Pocillopora damicornis, were exposed to 0 (controls), 1.0 (low) and 10 (moderate) µg l–1 diuron treatments for 2 to 3 mo prior to spawning or planulation. Diuron caused photoinhibition in each species, with pulse amplitude modulation (PAM) fluorometery recording consistent declines in effective quantum yields of 20% at 1.0 µg l–1 and 75% at 10 µg l–1 diuron compared to control corals. A. valida and P. damicornis were both sensitive to chronic diuron-induced photoinhibition, becoming severely bleached (loss of Symbiodinium spp. and/or reductions in light-harvesting pigments), especially at 10 µg l–1 diuron. At this moderate concentration, A. valida sustained both partial and full colony mortality. A. tenuis was more resistant to these concentrations of diuron, and neither bleached nor sustained partial mortality in any of the treatments. We found 2.5- to 5-fold reductions in total lipid content (coral tissue, oocytes and planulae) for the 3 species in the presence of diuron, indicating significant use of storage lipid to meet nutritional demands under conditions of chronic photoinhibition. Polyp fecundity was reduced by 6-fold in A. valida and both A. valida and P. damicornis were unable to spawn or planulate following long-term exposures to 10 µg l–1 diuron. This is the first study to investigate the sub-lethal effects of chronic herbicide-induced photoinhibition on symbiotic corals and provides evidence of a link between reduced energy acquisition due to PSII photoinhibition and reduced reproductive output in zooxanthellate corals.

Integrins are involved in a wide variety of cell adhesion processes, and have roles in gamete binding and fusion in mammals. Integrins have been also discovered in the scleractinian coral Acropora millepora (Cnidaria: Anthozoa). As a first step toward understanding the molecular basis of fertilization in corals, we examined the effect of polyclonal antisera raised against recombinant coral integrins on gamete interactions in A. millepora. Antiserum raised against integrin betacn1 dramatically decreased the binding of Acropora sperm to eggs and significantly decreased fertilization rates relative to preimmune serum and seawater controls. However, the antiserum against AmIntegrin alpha1 did not affect significantly either sperm-egg binding or fertilization. One possible explanation for this is that AmIntegrin alpha1 may preferentially mediate interactions with RGD-containing ligands, whereas mammalian alpha6 integrin (which is most directly implicated in gamete interactions) preferentially interacts with laminin-related ligands. Our results suggest that beta1 type integrins are involved in the fertilization process in Acropora and that some functions of these molecules may have been conserved between corals and mammals.

across the globe, we are witnessing the decline of coral reef ecosystems. One relatively new factor contributing to this decline is the outbreak of destructive infectious diseases, especially on Caribbean reefs. As the Coral Disease Working Group of the Coral Reef Targeted Research Program, our research focuses on four priorities: (1) assessing the global prevalence of coral disease, (2) investigating the environmental drivers of disease, (3) identifying the pathogens that cause disease, and (4) evaluating the coral’s ability to resist disease. Monitoring has revealed new coral-disease syndromes at each of four Global Environmental Fund Centers of Excellence: the Caribbean, the Philippines, Australia, and East Africa. Over the last 20 years, drastic (> 50 percent) loss of coral cover has occurred on the Yucatan Peninsula, even in pristine areas. Global surveys have revealed significant levels of disease and disease outbreaks occurring not only in the Caribbean “hotspots,” but also in sites throughout the Pacific and Indian Oceans. By monitoring coral disease, we will create a baseline and long-term data set that can be used to test specific hypotheses about how climate and anthropogenic drivers, such as decreasing water quality, threaten coral reef sustainability. One such hypothesis is that high-temperature anomalies drive outbreaks of disease by hindering the coral’s ability to fi ght infection and by increasing the pathogens’ virulence. We observed recurrent outbreaks following the warm summer months of two of the most damaging diseases in the Caribbean. In addition, we found that coral disease in the Great Barrier Reef correlated with warm temperature anomalies. In the Caribbean and Mediterranean Seas, virulence of known coral pathogens and the normal coral flora changed during high-temperature periods. Other stresses such as high nutrients and sedimentation may similarly alter the balance between the coral and its resident microbial flora.

Rates of progression and transmission of black band disease (BBD) on the staghorn coral, Acropora muricata, were compared between months for seasonal in situ studies and between temperature treatments in experimental aquaria manipulations at Lizard Island on the Great Barrier Reef (GBR). In situ field experiments demonstrated that BBD progressed along branches approximately twice as fast (1.7–2.4 times) during the austral summer month of January (0.99 ± 0.04 cm/day) than in the cooler months of July (0.58 ± 0.04 cm/day) and May (0.41 ± 0.07 cm/day). Transmission of BBD between colonies was also accelerated in warmer months, with signs of infection becoming visible 1.2 days earlier in January compared to May. The greater seawater temperatures by ∼2 to 3°C and light intensities by up to 650 μE/m2/s in January, suggest that rates of progression and transmission of BBD are linked to one or both of these parameters. Manipulative experiments in summer provide corroborative evidence that elevated temperatures increase rates of BBD progression, with the disease progressing 1.3 times more rapidly in the 32°C elevated temperature treatment than in the 30°C ambient treatment (1.17 ± 0.06 cm/day versus 0.92 ± 0.07 cm/day; F2,6 = 7.66, P = 0.022). In contrast, although a trend for greatest BBD progression was measured in elevated temperature treatments of 29°C (0.46 ± 0.07 cm/day) and 31°C (0.52 ± 0.06 cm/day) in winter, these rates did not differ significantly (F3,7 = 1.72, P = 0.249) from those measured for the ambient 27°C treatment (0.37 ± 0.06 cm/day) or the field controls (0.41 ± 0.09 cm/day). The lower rates of BBD progression in the 31°C winter treatment (0.52 ± 0.06 cm/day) than in the 30°C (0.92 ± 0.07 cm/day) summer treatment, may have been a response to 28-fold decreased light irradiance in the former, suggesting that high irradiance in combination with elevated temperatures may promote progression of BBD. Results from this study indicate that the impact of elevated temperature on BBD progression is complex with a combination of environmental factors including temperature and light playing key roles in progression and transmission of the disease.

Within populations of brooding sessile corals, sperm dispersal constitutes the mechanism by which gametes interact and mating occurs, and forms the first link in the network of processes that determine specieswide connectivity patterns. However, almost nothing is known about sperm dispersal for any internally fertilizing coral. In this study, we conducted a parentage analysis on coral larvae collected from an area of mapped colonies, to measure the distance sperm disperses for the first time in a reef-building coral and estimated the mating system characteristics of a recently identified putative cryptic species within the Seriatopora hystrix complex (ShA; Warner et al. 2015). We defined consensus criteria among several replicated methods (COLONY 2.0, CERVUS 3.0, MLTR v3.2) to maximize accuracy in paternity assignments. Thirteen progeny arrays indicated that this putative species produces exclusively sexually derived, primarily outcrossed larvae (mean t(m) = 0.999) in multiple paternity broods (mean r(p) = 0.119). Self-fertilization was directly detected at low frequency for all broods combined (2.8%), but comprised 23% of matings in one brood. Although over 82% of mating occurred between colonies within 10 m of each other (mean sperm dispersal = 5.5 m ± 4.37 SD), we found no evidence of inbreeding in the established population. Restricted dispersal of sperm compared to slightly greater larval dispersal appears to limit inbreeding among close relatives in this cryptic species. Our findings establish a good basis for further work on sperm dispersal in brooding corals and provide the first information about the mating system of a newly identified and abundant cryptic species.

The term white syndromes (WS) applies to irregular patterns of tissue loss exposing bands or patches of bare white skeleton. There are many different white diseases in the Caribbean, each with potentially different causative agents, there are likely to be many different types of white syndromes causing tissue loss on Indo-Pacific corals. Coral disease diagnosis and nomenclature are predominantly based on underwater observations of macroscopic characteristics, such as lesion color, lesion progression rate, and patterns and extent of tissue loss and exposed coral skeleton. Variability in the presence of bacteria associated with WS lesions has also been found in ultrastructural studies. Robust ecological assessments and rapidly developing molecular biological and chemical technologies all aimed at understanding the interactions between the agent, host and the environment, which all play a role in the etiology of white syndromes of the Indo-Pacific must be considered in determining causation.

Emerging infectious diseases are contributing to global declines in coral reef ecosystems, highlighting a growing need for aetiological knowledge to develop effective management strategies. In this review, we focus on black band disease (BBD), one of the most virulent diseases and the only polymicrobial disease so far known to affect corals. A multipartite microbial consortium dominated by Cyanobacteria, but also including sulfur-cycling bacteria, other bacterial groups and members of the Archaea and Eukarya, forms a sulfide-rich anaerobic mat that migrates across the surface of coral colonies, killing the underlying tissues. The polymicrobial nature of the disease challenges classic aetiological approaches to unravelling disease causation. Here, we synthesize current knowledge on the range of pathogens forming the microbial consortium with recent studies on the transmission, biogeochemistry and environmental drivers of BBD to develop a conceptual model of BBD pathogenesis. The model illustrates how the development of BBD virulence factors is linked to a cascade of microbial community shifts and associated functional roles that progressively develop the microbial consortium from comparatively benign cyanobacterial patches to virulent BBD lesions. This review showcases how an approach that integrates multiple key aspects of the disease provides insights essential to elucidating the aetiology of BBD.

We investigated the effect of stressors associated with offshore reef-based tourist platforms on the coral immune system. Our findings suggest that stressors associated with platform-related activities, in synergy with seasonal factors, compromise the coral immune system, contributing to the higher coral disease prevalence observed at these locations., Unravelling the contributions of local anthropogenic and seasonal environmental factors in suppressing the coral immune system is important for prioritizing management actions at reefs exposed to high levels of human activities. Here, we monitor health of the model coral Acropora millepora adjacent to a high-use and an unused reef-based tourist platform, plus a nearby control site without a platform, over 7 months spanning a typical austral summer. Comparisons of temporal patterns in a range of biochemical and genetic immune parameters (Toll-like receptor signalling pathway, lectin–complement system, prophenoloxidase-activating system and green fluorescent protein-like proteins) among healthy, injured and diseased corals revealed that corals exhibit a diverse array of immune responses to environmental and anthropogenic stressors. In healthy corals at the control site, expression of genes involved in the Toll-like receptor signalling pathway (MAPK p38, MEKK1, cFos and ATF4/5) and complement system (C3 and Bf) was modulated by seasonal environmental factors in summer months. Corals at reef platform sites experienced additional stressors over the summer, as evidenced by increased expression of various immune genes, including MAPK p38 and MEKK1. Despite increased expression of immune genes, signs of white syndromes were detected in 31% of study corals near tourist platforms in the warmest summer month. Evidence that colonies developing disease showed reduced expression of genes involved in the complement pathway prior to disease onset suggests that their immune systems may have been compromised. Responses to disease and physical damage primarily involved the melanization cascade and GFP-like proteins, and appeared to be sufficient for recovery when summer heat stress subsided. Overall, seasonal and anthropogenic factors may have interacted synergistically to overwhelm the immune systems of corals near reef platforms, leading to increased disease prevalence in summer at these sites.

International audience; Rising sea temperatures are likely to increase the frequency of disease outbreaks affecting reef-building corals through impacts on coral hosts and pathogens. We present and compare climate model projections of temperature conditions that will increase coral susceptibility to disease, pathogen abundance and pathogen virulence. Both moderate (RCP 4.5) and fossil fuel aggressive (RCP 8.5) emissions scenarios are examined. We also compare projections for the onset of disease-conducive conditions and severe annual coral bleaching, and produce a disease risk summary that combines climate stress with stress caused by local human activities. There is great spatial variation in the projections, both among and within the major ocean basins, in conditions favouring disease development. Our results indicate that disease is as likely to cause coral mortality as bleaching in the coming decades. These projections identify priority locations to reduce stress caused by local human activities and test management interventions to reduce disease impacts.

The importance of hybridization in the evolution of plant species is widely accepted, but its contributions to animal species evolution remain less recognized. Here we review evidence that hybridization has contributed to the evolution of reef corals, a group underpinning the coral reef ecosystem. Increasingly threatened by human and climate-related impacts, there is need to understand the evolutionary processes that have given rise to their diversity and contribute to their resilience. Reticulate evolutionary pathways among the ecologically prominent, mass-spawning genus Acropora suggest that hybridization, although rare on ecological timescales, has been instrumental in their diversification on evolutionary timescales. Evidence that coral hybrids colonize marginal habitats distinct from those of parental species' and that hybridization may be more frequent at peripheral boundaries of species' ranges supports a role for hybridization in range expansion and adaptation to changing environments. We conclude that outcomes of hybridization are significant for the future resilience of reef corals and warrant inclusion in conservation strategies.

We evaluated the role that demography may play in the formation of local aggregations of Sinularia flexibilis (Quoy & Gaimard, 1833), a soft coral that commonly dominates inshore coral reefs of the Great Barrier Reef (GBR), Australia. Two populations on inshore reefs of the Palm Islands were censused once a year for 3 years, starting 10 mo after the extensive bleaching mortality in early 1998. Larger colonies became more prevalent over time; mean colony size increasing by 35%, from 276 cm2 in 1998 to 373 cm2 in 2000. Growth rates were size dependent, with smaller colonies growing proportionally faster than larger colonies. Change in size relative to initial size indicated an expected mean annual growth of 128 cm2 for a 50-cm2 colony. Zero growth was predicted at 532±21cm2, with colonies larger than this likely to undergo fission or shrink. Forty-three percent of colonies were undergoing fission at any time at both localities. Most new colonies were produced by fission (70%, n=285), with the remainder produced by the recruitment of sexually produced larvae (19%) or by colony translocation (11%). The sexual and asexual recruitment rates were 0.24 and 1.0 recruits m- 2 year−1, respectively. Opportunistic recruitment and rapid growth following disturbances are commonly assumed to be the mechanisms leading soft corals to dominate locally. In this study, these mechanisms operated more slowly than expected, with no net change in population size.

Symbiotic relationships have contributed greatly to the evolution and maintenance of biological diversity. On the Great Barrier Reef, species of obligate coral-dwelling fishes (genus Gobiodon) coexist by selectively recruiting to colonies of Acropora nasuta that differ in branch-tip colour. In this study, we investigate genetic variability among sympatric populations of two colour morphs of A. nasuta ('blue-tip' and 'brown-tip') living in symbiosis with two fish species, Gobiodon histrio and G. quinquestrigatus, respectively, to determine whether gobies are selecting between intraspecific colour polymorphisms or cryptic coral species. We also examine genetic differentiation among coral populations containing both these colour morphs that are separated by metres between local sites, tens of kilometres across the continental shelf and hundreds of kilometres along the Great Barrier Reef. We use three nuclear DNA loci, two of which we present here for the first time for Acropora. No significant genetic differentiation was detected between sympatric colour morphs at these three loci. Hence, symbiotic gobies are selecting among colour morphs of A. nasuta, rather than cryptic species. Significant genetic geographical structuring was observed among populations, independent of colour, at regional (i.e. latitudinal separation by < 500 km) and cross-shelf (< 50 km) scales, alongside relative homogeneity between local populations on within reef scales (< 5 km). This contrasts with the reported absence of large-scale genetic structuring in A. valida, which is a member of the same species group as A. nasuta. Apparent differences in biogeographical structuring between species within the A. nasuta group emphasize the need for comparative sampling across both spatial (i.e. within reefs, between reefs and between regions) and taxonomic scales (i.e. within and between closely related species).

In colonial modular organisms, differences in module size and colony growth patterns among species have the potential to impose varying constraints on reproductive investment. Here, we compare reproductive output among seven morphologically different species of spawning reef corals, and analyse the relationship between reproductive output and module (polyp) size. Reproductive output ranged between 132 and 384 J cm(-2), with lipid constituting the key indicator of energy investment. Lipid decreased by 85-100%, whereas protein and carbohydrate were relatively invariant between pre- and post-spawning tissues in all species, representing 1-15% and1%, respectively, of the energy investment to reproductive output. The ratio of energy content in reproductive to somatic tissues (gonadosomatic index, GSI) varied among species from 0.20 (Symphyllia recta) to 1.31 (Acropora tenuis), the latter being the highest value reported for any iteroparous marine invertebrate. Surprisingly, small-polyped species (Acropora, Montipora) had 2- to 6-fold higher GSIs than large-polyped ones (Lobophyllia, Symphyllia). Energy equivalents of tissues increased with the 1.50-1.76 power of polyp diameter for somatic tissues and with the 1.42-1.80 power of polyp diameter for reproductive output. In both cases, increases in energy equivalents with polyp diameter were less than the scaling exponent of 3 predicted for an isometric relationship between tissue volume (or mass) and polyp diameter, indicating significant constraints of space, design or physiological energetics with increasing polyp size. We hypothesise that such constraints have played a key role in the evolution of modularity in cnidarians.

Species boundaries among five sympatric coral species of the Indo-Pacific Acropora aspera group were examined by a combination of in vitro breeding trials, comparisons of spawning times and DNA sequence analysis of ribosomal DNA internal transcribed spacer (rDNA ITS) and 5.8S regions. The breeding trials showed that reproductive compatibility exists between at least some colonies of all the species pairs tested, suggesting a large potential for natural hybridization and introgression. The Acropora ITS regions exhibited extremely high levels of variability (up to approximately 62% for ITS1, approximately 11% for 5.8S and approximately 43% for ITS2), but most of the variation was shared among four of the five species, A. millepora, A. papillare, A. pulchra and A. spathulata, consistent with extensive introgression. Phylogenetic analyses did not resolve these four species as distinct clusters across a wide biogeographic region stretching from the southern Great Barrier Reef to Papua New Guinea. However, most colonies of the fifth species, A. aspera, constituted a distinct clade in phylogenetic analyses. This is consistent with our observations of a semi-permeable temporal barrier involving differences in spawning times between this and the other four species. Although the majority of colonies of all five species generally spawned within 90 min of each other, in two out of four years, gametes were absent prior to mass spawning episodes from at least some A. aspera colonies. Hence, our data suggest that transient reproductive barriers may be the result of year-to-year variation in the date of spawning and that this difference in spawning time contributes to the genetic structure detected among Acropora species in this group. Occasional leakage through the reproductive barrier was confirmed by the observation of A. aspera xA. pulchra F1 hybrids, identified based on additivity of ITS sequences.

A major challenge for understanding the evolutionary genetics of mass-spawning corals is to explain the maintenance of discrete morphospecies in view of high rates of interspecific fertilization in vitro and nonmonophyletic patterns in molecular phylogenies. In this study, we focused on Acropora cytherea and A. hyacinthus, which have one of the highest potentials for interspecific fertilization. Using sequences of a nuclear intron, we performed phylogenetic and nested clade analyses (NCA). Both species were polyphyletic in molecular phylogenies, but the NCA indicated that they constitute statistically distinguishable lineages. Phylogenetic analysis using an intergenic region of the mitochondrial DNA (mtDNA), was inconclusive because of low levels of variability in this marker. The position of these two species differed between the nuclear DNA (nDNA) and mtDNA phylogenies and was also at odds with a cladistic analysis based on morphology. We conclude that despite the potential for high levels of hybridization and introgression, A. cytherea and A. hyacinthus constitute statistically distinguishable lineages and their taxonomic status is consistent with the cohesion species concept.

High cross-fertilization rates in vitro and non-monophyletic patterns in molecular phylogenies challenge the taxonomic status of species in the coral genus Acropora. We present data from eight polymorphic allozyme loci that indicate small, but significant, differentiation between sympatric populations of Acropora cytherea and Acropora hyacinthus (F(ST) = 0.025-0.068, p < 0.05), a pair of acroporid corals with very high interspecific fertilization rates in vitro. Although no fixed allelic differences were found between these species, the absence of genetic differentiation between widely allopatric populations suggests that allele frequency differences between A. cytherea and A. hyacinthus in sympatry are biologically significant. By contrast, populations of Acropora tenuis, a species which spawns 2-3 hours earlier and shows low cross-fertilization rates with congeners in vitro, were clearly distinct from A. cytherea and A. hyacinthus (F(ST) = 0.427-0.465, p < 0.05). Moreover, allopatric populations of A. tenuis differed significantly, possibly as a consequence of its relatively short period of larval competency. Our results effectively rule out the possibility that A. hyacinthus and A. cytherea are morphotypes within a single species, and indicate that hybridization occurs relatively infrequently between these taxa in nature.

Regional-scale variation of recruitment by marine organisms may reflect geographic patterns in adult stock sizes or fecundities, large-scale hydrodynamic features that influence the transport of larvae (e.g., currents, upwelling), and patterns of early mortality. In turn, recruitment may play a vital role in determining patterns of adult abundance and community structure, from local to biogeographic scales. We examined spatial variation in recruitment by corals at a regional scale, along 3300 km of the tropical and subtropical coast of eastern Australia (10°–31° S). We used two complementary approaches: (1) a metaanalysis of 21 different studies undertaken over a 16-yr period, each of which was generally conducted at a single reef, and (2) a large-scale sampling effort in which recruitment was measured in two years on 33 reefs arrayed along the length of the Great Barrier Reef (GBR). Our goal is to compare the emergent large-scale picture derived from many small-scale studies with patterns revealed by shorter-term regional sampling. The two approaches show very similar large-scale patterns. Recruitment by spawning corals (mainly acroporids) was highest in the central GBR and declined steadily with increasing latitude by up to more than 20-fold. A smaller decline occurred on the northern GBR between Australian and Papua New Guinea. Recruitment by brooding corals (mostly pocilloporids) was greatest in the northern GBR and also declined to the south. The latitudinal decline in brooders was three-to fivefold, i.e., not as great as for spawners. Consequently, the proportion of brooded recruits increased to the south, and they generally exceeded spawners on the southern GBR and on isolated subtropical reefs at higher latitudes. Our meta-analysis shows that fully half of the variation in the ratio of spawners to brooders is attributable to one of 11 variables that we extracted from the published studies: the month when the recruitment panels were deployed. This result suggests that the intensity and timing of spawning have a crucial impact on large-scale patterns of recruitment. Elsewhere, we tested this hypothesis in the field, and confirmed that regional variation in recruitment by spawning acroporid corals was driven by spatial and temporal variation in the extent of mass spawning. Together, large-scale sampling and meta-analyses provide a powerful, combined approach for investigating large-scale patterns and the mechanisms underlying them.

While coral reefs are increasingly threatened worldwide, they are also increasingly used for recreational activities. Given the environmental and socio-economic significance of coral reefs, understanding the links between human activities and coral health and evaluating the efficacy of marine protected areas (MPAs) as a management regime to prevent further deterioration are critically important. The aim of this study was to quantify indicators of coral health at sites inside and outside a newly rezoned MPA framework in the dive tourism hotspot of Koh Tao, Thailand. We found that patterns in the health and diversity of coral communities one year on did not reflect the protected status conferred by newly zoned MPAs, but instead reflected past history of recreational use around the island. Sites characterised as past high-use sites had lower mean percent cover of hard corals overall and of corals in the typically disease- and disturbance-susceptible family Acroporidae, but higher mean cover of species in the more weedy family Agariciidae. Past high use sites also had higher mean prevalence of infectious diseases and other indicators of compromised health. Sites within the newly established MPAs are currently subjected to higher levels of environmental and anthropogenic pressures, with sedimentation, algal overgrowth, feeding scars from Drupella snails, and breakage particularly prevalent compared to sites in non-MPA areas. Given the greater prevalence of these factors within protected sites, the capacity of the MPA framework to effectively prevent further deterioration of Koh Tao's reefs is unclear. Nevertheless, our study constitutes a strong baseline for future long-term evaluations of the potential of MPAs to maintain coral health and diversity on highly threatened reefs.

Increasing physical damage on coral reefs from predation, storms and anthropogenic disturbances highlights the need to understand the impact of injury on the coral immune system. In this study, we examined the regulation of the coral immune response over 10 days following physical trauma artificially inflicted on in situ colonies of the coral Acropora aspera, simultaneously with bacterial colonization of the lesions. Corals responded to injury by increasing the expression of immune system-related genes involved in the Toll-like and NOD-like receptor signalling pathways and the lectin-complement system in three phases (

The bacterium Vibrio coralliilyticus has been implicated as the causative agent of coral tissue loss diseases (collectively known as white syndromes) at sites across the Indo-Pacific and represents an emerging model pathogen for understanding the mechanisms linking bacterial infection and coral disease. In this study, we used a mini-Tn7 transposon delivery system to chromosomally label a strain of V. coralliilyticus isolated from a white syndrome disease lesion with a green fluorescent protein gene (GFP). We then tested the utility of this modified strain as a research tool for studies of coral host–pathogen interactions. A suite of biochemical assays and experimental infection trials in a range of model organisms confirmed that insertion of the GFP gene did not interfere with the labeled strain’s virulence. Using epifluorescence video microscopy, the GFP-labeled strain could be reliably distinguished from non-labeled bacteria present in the coral holobiont, and the pathogen’s interactions with the coral host could be visualized in real time. This study demonstrates that chromosomal GFP labeling is a useful technique for visualization and tracking of coral pathogens and provides a novel tool to investigate the role of V. coralliilyticus in coral disease pathogenesis., Human Frontier Science Program (Strasbourg, France) (No. RGY0089RS)

The capacity of reef-building corals to associate with environmentally-appropriate types of endosymbionts from the dinoflagellate genus Symbiodinium contributes significantly to their success at local scales. Additionally, some corals are able to acclimatize to environmental perturbations by shuffling the relative proportions of different Symbiodinium types hosted. Understanding the dynamics of these symbioses requires a sensitive and quantitative method of Symbiodinium genotyping. Electrophoresis methods, still widely utilized for this purpose, are predominantly qualitative and cannot guarantee detection of a background type below 10% of the total Symbiodinium population. Here, the relative abundances of four Symbiodinium types (A13, C1, C3, and D1) in mixed samples of known composition were quantified using deep sequencing of the internal transcribed spacer of the ribosomal RNA gene (ITS-2) by means of Next Generation Sequencing (NGS) using Roche 454. In samples dominated by each of the four Symbiodinium types tested, background levels of the other three types were detected when present at 5%, 1%, and 0.1% levels, and their relative abundances were quantified with high (A13, C1, D1) to variable (C3) accuracy. The potential of this deep sequencing method for resolving fine-scale genetic diversity within a symbiont type was further demonstrated in a natural symbiosis using ITS-1, and uncovered reef-specific differences in the composition of Symbiodinium microadriaticum in two species of acroporid corals (Acropora digitifera and A. hyacinthus) from Palau. The ability of deep sequencing of the ITS locus (1 and 2) to detect and quantify low-abundant Symbiodinium types, as well as finer-scale diversity below the type level, will enable more robust quantification of local genetic diversity in Symbiodinium populations. This method will help to elucidate the role that background types have in maximizing coral fitness across diverse environments and in response to environmental change.

The role of microhabitat in structuring epifaunal communities on four corals of varying morphology in the genus Acropora (A. millepora, A. hyacinthus, A. pulchra, A. formosa) was determined on two fringing reefs in the central Great Barrier Reef. Greater abundance and species richness of epifauna on tightly branched coral species in comparison to their rarity or absence on open-branched species suggests that protection afforded by complex habitats is important in structuring coral epifaunal communities. Within species, neither total colony space nor live surface area of corals was correlated with either the abundance or species richness of associated epifauna. However, space between branches significantly affected the size of Tetralia crabs associated with different coral species. Patterns in the size distribution of Tetralia on two species of Acropora suggest that crabs select coral hosts according to branch spacing, changing host species as they grow larger.

Experimental bleaching reduces the levels of important biochemical parameters in adult tissues and eggs of the soft coral Lobophytum compactum. Protein, lipid, mycosporine-like amino acids (MAAs) and carotenoid concentrations remained lower in bleached adults than in controls for at least 8 months. Reductions in concentrations of all four parameters were greater in eggs than in maternal tissues, potentially jeopardizing egg and larval viability. In particular, reductions in lipids, proteins and carotenoids in tissues of heavily bleached soft corals were amplified approximately twofold in eggs. In comparison, amplification of maternal tissue reductions were not as great for MAAs, suggesting that MAAs are given higher priority in egg provisioning. Our finding that MAA levels are normally three times higher in eggs than in unbleached maternal tissues supports the importance of MAAs for larval survival. Twenty months after experimental bleaching the biochemical composition of both adult tissues and their eggs were indistinguishable from those of control (unbleached) soft corals.

We document long-term effects of a simulated bleaching event on the reproductive output and offspring viability of the soft coral Lobophytum compactum. Corals were subjected to temperature and solar radiation treatments to produce both moderately (48–60%) and heavily (90–95%) bleached colonies. Although bleached colonies recovered their zooxanthellae within 10 to 18 weeks, impacts on reproductive output were significant for at least two annual spawning seasons. In the first year, both polyp fecundity and mean oocyte diameter were reduced and inversely correlated with the degree of bleaching, with complete failure of fertilization in the group of heavily bleached colonies. For moderately bleached soft corals, survival and growth of sexual offspring did not differ significantly from those of unbleached colonies. Although no further reductions in zooxanthellae densities in experimental soft corals were recorded throughout the subsequent second year, egg size and fecundity of the heavily bleached soft corals were still significantly reduced 20 months later. Severe bleaching clearly has long-term sub-lethal impacts, reducing overall reproductive output for at least two spawning seasons.

Although Acropora is the most species-rich genus of the scleractinian (stony) corals, only three species occur in the Caribbean: A. cervicornis, A. palmata and A. prolifera. Based on overall coral morphology, abundance and distribution patterns, it has been suggested that A. prolifera may be a hybrid between A. cervicornis and A. palmata. The species boundaries among these three morphospecies were examined using DNA sequence analyses of the nuclear Pax-C 46/47 intron and the ribosomal DNA Internal Transcribed Spacer (ITS1 and ITS2) and 5.8S regions. Moderate levels of sequence variability were observed in the ITS and 5.8S sequences (up to 5.2% overall sequence difference), but variability within species was as large as between species and all three species carried similar sequences. Since this is unlikely to represent a shared ancestral polymorphism, the data suggest that introgressive hybridization occurs among the three species. For the Pax-C intron, A. cervicornis and A. palmata had very distinct allele frequencies and A. cervicornis carried a unique allele at a frequency of 0.769 (although sequence differences between alleles were small). All A. prolifera colonies examined were heterozygous for the Pax-C intron, whereas heterozygosity was only 0.286 and 0.333 for A. cervicornis and A. palmata, respectively. These data support the hypothesis that A. prolifera is the product of hybridization between two species that have a different allelic composition for the Pax-C intron, i.e. A. cervicornis and A. palmata. We therefore suggest that A. prolifera is a hybrid between A. cervicornis and A. palmata, which backcrosses with the parental species at low frequency.

"Supply-side" ecology recognizes the potential role that recruitment plays in the local population dynamics of open systems. Apart from the applied fisheries literature, the converse link between adults and the production of cohorts of recruits has received much less attention. We used a hierarchical sampling design to investigate the relationships between adult abundance, fecundity, and rates of larval recruitment by acroporid corals on 33 reefs in five sectors (250-400 km apart) stretching from north to south along the length of the Great Barrier Reef, Australia. Our goal was to quantify patterns of recruitment at multiple scales, and to explore the underlying mechanisms. Specifically, we predicted that large-scale patterns of recruitment could be driven by changes in the abundance of adults and/or their fecundity, i.e., that corals exhibit a stock-recruitment relationship. The amount of recruitment by acroporids in each of two breeding seasons varied by more than 35-fold among the five sectors. Adult density varied only twofold among sectors and was not correlated with recruitment at the sector or reef scale. In contrast, fecundity levels (the proportion of colonies on each reef that contained ripe eggs) varied from 15% to 100%, depending on sector, year, and species. Spatial and temporal variation in the fecundity of each of three common Acropora species explained most of the variation (72%) in recruitment by acroporids, indicating that the production of larvae is a major determinant of levels of recruitment at large scales. Once fecundity was accounted for, none of the other variables we examined (sector, reef area, abundance of adults, or year) contributed significantly to variation in recruitment. The relationship between fecundity and recruitment was nonlinear, i.e., rates of recruitment increased disproportionately when and where the proportion of gravid colonies approached 100%. This pattern is consistent with the hypothesis that enhanced fertilization success and/or predator satiation occurs during mass-spawning events. Furthermore, it implies that small, sublethal changes in fecundity of corals could result in major reductions in recruitment.

Experimental studies of the upper thermal limits of corals from Orpheus Island, an inshore reef in the central Great Barrier Reef, show that Acropora formosa has a 5-day 50%-bleaching threshold of between 31 and 32 °C in summer, only 2 to 3 °C higher than local mean summer temperatures (29 °C). Summer bleaching thresholds for Pocillopora damicornis and A. elseyi were 1 °C higher (between 32 and 33 °C). The winter bleaching threshold of Pocillopora damicornis was 1 °C lower than its summer threshold, indicating that seasonal acclimatisation may take place. This seasonal difference raises the possibility that at least some corals may be capable of short-term thermal acclimatisation. Neither P. damicornis nor A. elseyi showed habitat-specific (reef flat versus reef slope) differences in bleaching thresholds. Further, colonies of P. damicornis collected from sites 3 km apart also showed no difference in bleaching threshold despite populations of this species responding differently at these two sites during a natural bleaching event. The bleaching thresholds determined in this study are best considered as the maximum tolerable temperatures for local populations of these species because they were determined in the absence of additional stressors (e.g. high light) which often co-occur during natural bleaching events. We consider the 5-day 50% bleaching thresholds determined in these experiments to be fair indicators of upper thermal limits, because >50% of a sample population died when allowed to recover in situ. We found a delay of up to a month in the bleaching response of corals following thermal stress, a result that has implications for identifying the timing of stressful conditions in natural bleaching events.

Synchronous spawning of many coral species that co-occur on Indo-Pacific reefs raises the possibility that hybridization plays a role in their evolution. Here we use experimental crosses to examine mating compatibilities and breeding barriers in a group of sessile animals whose mating systems are primarily governed by interactions among free-spawned gametes. We found hybridization occurs readily in more than one-third of 42 species pairs from the common genera Acropora, Montipora and Platygyra. Mean fertilization success ranged from 1% to 50% in species crosses, but standard deviations about these means were large and in some cases, fertilization success in individual colony matings was greater than 95%. Cases of high fertility in individual, interspecific matings were found in all three genera. Hybridization occurred most readily between species that were morphologically similar, identifying areas where current taxonomic judgements may require further testing. However, cases of significant hybridization also occurred between species that are morphologically distinct. Evidence of in vitro hybridization combined with the lack of either temporal or spatial barriers to interbreeding among field populations of these species, indicates that natural hybridization may occur commonly between congeneric corals that are currently recognized as distinct species. We also detected mating incompatibilities between some colonies within some species. In some cases, incompatible colonies corresponded to distinct morphotypes, but not in others. Thus some breeding groups in scleractinian corals are potentially larger, but others are smaller, than would be predicted using morphological criteria. Gamete incompatibilities within a morphospecies that readily hybridizes with other species may be the result of a mating system that is governed by gamete-level interactions. Imprecision in the alignment of morphological and breeding boundaries suggests a single species concept may not apply to scleractinian corals and challenges the tacit assumption that currently defined coral species encompass biological, evolutionary and phylogenetic species. Hybridization between supposedly isolated species introduces a reticulate nature to the evolution of corals and has profound implications for present understanding of the population genetics, phylogenetics, and evolutionary biology of scleractinian corals.

Black band disease (BBD) is a microbial consortium that creates anoxic, sulfide-rich microenvironments and kills underlying coral tissues as it rapidly migrates across colonies. Although bacterial communities associated with BBD have been studied extensively, the presence and roles of archaea are unexplored. Using amplicon-pyrosequencing of 16S ribosomal RNA genes, we investigated the community structure of both archaea and bacteria within microbial lesions of BBD and the less-virulent precursor stage, 'cyanobacterial patches' (CP), affecting the coral Montipora hispida. We detected characteristic shifts in microbial communities during the development of BBD from CP, reflecting microenvironmental changes within lesions. Archaeal profiles in CP suggested a diverse assemblage affiliated with the Thaumarchaeota and Euryarchaeota, similar to communities described for oxic marine environments. In contrast, a novel ribotype, distantly affiliated to the Euryarchaeota, dominated up to 94% of archaeal sequences retrieved from BBD. The physiological characteristics of this dominant archaeal ribotype are unknown because of the novelty of its 16S ribosomal RNA gene sequences; however, their prominent associations with BBD lesions suggest the ability to thrive in the organic- and sulfide-rich anoxic microenvironment characteristic of BBD lesions. Discovery of this novel archaeal ribotype provides new insights into the microbial ecology and aetiology of BBD.