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Tropical reef-building corals exist in intimate symbiosis with diverse microbes and viruses. Coral microbiomes are generally much less diverse than their environment, but across studied corals, the biodiversity of these microbiomes varies greatly. It has previously been hypothesized that differences in coral innate immunity in general, and the copy number of TIR-domain containing innate immune genes in particular, may drive interspecific differences in microbiome structure. Despite many existing studies of coral microbiomes, this hypothesis has previously been difficult to test due to a lack of consistently collected cross-species data on coral microbiomes. In this manuscript, we reannotate TIR-domain containing genes across diverse coral genomes, and use phylogenetic comparative methods to compare these innate immune gene copy numbers against 16S rRNA marker gene data on coral mucus, tissue, and skeleton microbiomes from the Global Coral Microbiome Project (GCMP). The copy number of Toll-like receptor (TLRs) and Interleukin-1 receptor (IL-1Rs) gene families, as well as the total genomic count of their constituent domains (LRR and TIR domains; and Ig and TIR domains, respectively), explained most interspecific differences in microbiome richness and beta-diversity among corals with sequenced genomes. We find that these correlations are also anatomically specific, with an especially strong correlation between IL-1R gene copy numbers and microbiome richness in the coral’s endolithic skeleton. Together, these results suggest innate immunity may play a key role in sculpting microbiome structure in corals.

Evolutionary tradeoffs between life-history strategies are central to animal evolution. However, because microbes can influence aspects of host physiology, behavior, and resistance to stress or disease, changes in animal-microbial symbioses have the potential to mediate life-history tradeoffs. Scleractinian corals provide a highly biodiverse and data-rich host system to test this idea, made more relevant by increases in coral disease outbreaks as a result of anthropogenic changes to climate and reef ecosystems. Identifying factors that determine coral disease susceptibility has therefore become a focus for reef conservation efforts. Using a comparative approach, we tested if coral microbiomes correlate with disease susceptibility across 425 million years of coral evolution by combining a cross-species coral microbiome survey (the “Global Coral Microbiome Project”) with long-term disease prevalence data at multiple sites. Interpreting these data in their phylogenetic context, we show that microbial dominance and composition predict disease susceptibility. We trace this dominance-disease association to a single putatively beneficial bacterial symbiont,Endozoicomonas, whose relative abundance in coral tissue explained 30% of variation in disease susceptibility and 60% of variation in microbiome dominance across 40 coral genera. Conversely,Endozoicomonasabundances in coral tissue strongly correlated with high growth rates. These results demonstrate that the evolution of microbial symbiosis in corals correlates with both disease prevalence and growth rate. Exploration of the mechanistic basis for these findings will be important for our understanding of how microbial symbiosis influences animal life-history tradeoffs, and in efforts to use microbes to increase coral growth or disease resistancein-situ.Significance StatementThe evolution of tropical corals, like that of many organisms, involves tradeoffs in life-history strategy. We sought to test whether microbes influence coral life-history traits. Comparative data from a census of modern coral microbes, combined with long term disease surveys in three regions, provide evidence for a correlation between microbiome structure, growth rate, and disease susceptibility during coral evolution. These trends were driven primarily by changes in the relative abundance ofEndozoicomonasin coral tissue microbiomes, suggesting the novel hypothesis thatEndozoicomonasmay allow corals to grow more quickly at the cost of greater vulnerability to disease. Thus, symbiosis with microbes may be an important aspect of animal life-history strategy.

Corals are dependent upon lipids as energy reserves to mount a metabolic response to biotic and abiotic challenges. This study profiled lipids, fatty acids, and microbial communities of healthy and white syndrome (WS) diseased colonies of Acropora hyacinthus sampled from reefs in Western Australia, the Great Barrier Reef, and Palmyra Atoll. Total lipid levels varied significantly among locations, though a consistent stepwise decrease from healthy tissues from healthy colonies (HH) to healthy tissue on WS-diseased colonies (HD; i.e. preceding the lesion boundary) to diseased tissue on diseased colonies (DD; i.e. lesion front) was observed, demonstrating a reduction in energy reserves. Lipids in HH tissues were comprised of high energy lipid classes, while HD and DD tissues contained greater proportions of structural lipids. Bacterial profiling through 16S rRNA gene sequencing and histology showed no bacterial taxa linked to WS causation. However, the relative abundance of Rhodobacteraceae-affiliated sequences increased in DD tissues, suggesting opportunistic proliferation of these taxa. While the cause of WS remains inconclusive, this study demonstrates that the lipid profiles of HD tissues was more similar to DD tissues than to HH tissues, reflecting a colony-wide systemic effect and provides insight into the metabolic immune response of WS-infected Indo-Pacific corals. This project was funded in part by Earthwatch Institute and Mitsubishi Corporation. Scopus

Coral reef restoration is a rapidly growing movement galvanized by the accelerating degradation of the world's tropical coral reefs. The need for concerted and collaborative action focused on the recovery of coral reef ecosystems coalesced in the creation of the Coral Restoration Consortium (CRC) in 2017. In March 2020, the CRC leadership team met for a biennial review of international coral reef restoration efforts and a discussion of perceived knowledge and implementation bottlenecks that may impair scalability and efficacy. Herein we present six priorities wherein the CRC will foster scientific advancement and collaboration to: (1) increase restoration efficiency, focusing on scale and cost-effectiveness of deployment; (2) scale up larval-based coral restoration efforts, emphasizing recruit health, growth, and survival; (3) ensure restoration of threatened coral species proceeds within a population-genetics management context; (4) support a holistic approach to coral reef ecosystem restoration; (5) develop and promote the use of standardized terms and metrics for coral reef restoration; and (6) support coral reef restoration practitioners working in diverse geographic locations. These priorities are not exhaustive nor do we imply that accomplishing these tasks alone will be sufficient to restore coral reefs globally; rather these are topics where we feel the CRC community of practice can make timely and significant contributions to facilitate the growth of coral reef restoration as a practical conservation strategy. The goal for these collective actions is to provide tangible, local-scale advancements in reef condition that offset declines resulting from local and global stressors including climate change.

High-resolution benthic habitat data fill an important knowledge gap for many areas of the world and are essential for strategic marine conservation planning and implementing effective resource management. Many countries lack the resources and capacity to create these products, which has hindered the development of accurate ecological baselines for assessing protection needs for coastal and marine habitats and monitoring change to guide adaptive management actions. The PlanetScope (PS) Dove Classic SmallSat constellation delivers high-resolution imagery (4 m) and near-daily global coverage that facilitates the compilation of a cloud-free and optimal water column image composite of the Caribbean’s nearshore environment. These data were used to develop a first-of-its-kind regional thirteen-class benthic habitat map to 30 m water depth using an object-based image analysis (OBIA) approach. A total of 203,676 km2 of shallow benthic habitat across the Insular Caribbean was mapped, representing 5% coral reef, 43% seagrass, 15% hardbottom, and 37% other habitats. Results from a combined major class accuracy assessment yielded an overall accuracy of 80% with a standard error of less than 1% yielding a confidence interval of 78–82%. Of the total area mapped, 15% of these habitats (31,311.7 km2) are within a marine protected or managed area. This information provides a baseline of ecological data for developing and executing more strategic conservation actions, including implementing more effective marine spatial plans, prioritizing and improving marine protected area design, monitoring condition and change for post-storm damage assessments, and providing more accurate habitat data for ecosystem service models.

Over the past decade, coral restoration efforts have increased as reefs continue to decline at unprecedented rates. Identifying suitable coral outplanting locations to maximize coral survival continues to be one of the biggest challenges for restoration practitioners. Here, we demonstrate methods of using derivatives from imaging spectroscopy from the Global Airborne Observatory (GAO) to identify suitable coral outplant sites and report on the survival rates of restored coral at those sites. Outplant sites for a community-based, citizen science outplant event in Bávaro, Dominican Republic, were identified using expert-defined criteria applied to a suitability model from data layers derived from airborne imagery. Photo quadrat analysis of the benthic community confirmed the accuracy of airborne remote sensing maps with live coral cover averaging 3.5–4% and mean algal cover (macro algae and turf) ranging from 28 to 32%. Coral outplant sites were selected at 3–7 m depth with maximized levels of habitat complexity (i.e., rugosity) and live coral cover and minimized levels of macroalgal cover, as predicted by the imaging spectrometer data. In November 2019, 1,722 Acropora cervicornis fragments (80–180 mm in length) were outplanted to these sites. Surveys conducted in January 2020 in four of these sites confirmed that 92% of outplants survived after 3 months. By October 2020 (11 months after outplanting), survivorship remained above 76%. These results demonstrate higher than average success rates for coral outplant survival for this species. An online tool was developed to enable replication and facilitate future selection of coral restoration sites. Our objective is to present a case study that uses GAO-derived map products within a suitability model framework to provide a quantitative and replicable method for selecting coral restoration sites with the goal of increasing outplant survival over time.

Bacteria associated with coral hosts are diverse and abundant, with recent studies suggesting involvement of these symbionts in host resilience to anthropogenic stress. Despite their putative importance, the work dedicated to culturing coral-associated bacteria has received little attention. Combining published and unpublished data, here we report a comprehensive overview of the diversity and function of culturable bacteria isolated from corals originating from tropical, temperate, and cold-water habitats. A total of 3,055 isolates from 52 studies were considered by our metasurvey. Of these, 1,045 had full-length 16S rRNA gene sequences, spanning 138 formally described and 12 putatively novel bacterial genera across the Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria phyla. We performed comparative genomic analysis using the available genomes of 74 strains and identified potential signatures of beneficial bacterium-coral symbioses among the strains. Our analysis revealed >400 biosynthetic gene clusters that underlie the biosynthesis of antioxidant, antimicrobial, cytotoxic, and other secondary metabolites. Moreover, we uncovered genomic features-not previously described for coral-bacterium symbioses-potentially involved in host colonization and host-symbiont recognition, antiviral defense mechanisms, and/or integrated metabolic interactions, which we suggest as novel targets for the screening of coral probiotics. Our results highlight the importance of bacterial cultures to elucidate coral holobiont functioning and guide the selection of probiotic candidates to promote coral resilience and improve holistic and customized reef restoration and rehabilitation efforts. IMPORTANCE Our paper is the first study to synthesize currently available but decentralized data of cultured microbes associated with corals. We were able to collate 3,055 isolates across a number of published studies and unpublished collections from various laboratories and researchers around the world. This equated to 1,045 individual isolates which had full-length 16S rRNA gene sequences, after filtering of the original 3,055. We also explored which of these had genomes available. Originally, only 36 were available, and as part of this study, we added a further 38-equating to 74 in total. From this, we investigated potential genetic signatures that may facilitate a host-associated lifestyle. Further, such a resource is an important step in the selection of probiotic candidates, which are being investigated for promoting coral resilience and potentially applied as a novel strategy in reef restoration and rehabilitation efforts. In the spirit of open access, we have ensured this collection is available to the wider research community through the web site http://isolates.reefgenomics.org/ with the hope many scientists across the globe will ask for access to these cultures for future studies. published

Scleractinian corals’ microbial symbionts influence host health, yet how coral microbiomes assembled over evolution is not well understood. We survey bacterial and archaeal communities in phylogenetically diverse Australian corals representing more than 425 million years of diversification. We show that coral microbiomes are anatomically compartmentalized in both modern microbial ecology and evolutionary assembly. Coral mucus, tissue, and skeleton microbiomes differ in microbial community composition, richness, and response to host vs. environmental drivers. We also find evidence of coral-microbe phylosymbiosis, in which coral microbiome composition and richness reflect coral phylogeny. Surprisingly, the coral skeleton represents the most biodiverse coral microbiome, and also shows the strongest evidence of phylosymbiosis. Interactions between bacterial and coral phylogeny significantly influence the abundance of four groups of bacteria–including Endozoicomonas-like bacteria, which divide into host-generalist and host-specific subclades. Together these results trace microbial symbiosis across anatomy during the evolution of a basal animal lineage., Associations between corals and symbiotic microorganisms could be driven by the environment or shared evolutionary history. Here, the authors examine relationships between coral phylogenies and associated microbiomes, finding evidence of phylosymbiosis in microbes from coral skeleton and tissue, but not mucus.

Coral photosynthetic endosymbionts (Symbiodinium) are phylogenetically very diverse, yet the extent of inter- and intraspecific functional variation within clades remains largely underexplored. Understanding this variability will be critical for future research on climate change mediated responses. A properly functioning thylakoid membrane is essential for optimal photosynthetic performance both in free living and in hospite conditions. Here, we analyse the thylakoid membrane melting points of 13 Symbiodinium strains from species in Clades B and A, grown at both control (26 °C) and high temperature (31 °C). We observed a broad range of responses to thermal stress regardless of taxonomic rank. Our results support and augment a growing body of the literature demonstrating that functional differences among Symbiodinium spp. are as distinct at lower taxonomic levels (i.e. interspecific) as they are among major clades. These findings highlight the importance of assessing the variability of plastid traits across the Symbiodinium tree.

Bacterial diversity associated with corals has been studied extensively, however, localization of bacterial associations within the holobiont is still poorly resolved. Here we provide novel insight into the localization of coral-associated microbial aggregates (CAMAs) within tissues of the coral Acropora hyacinthus. In total, 318 and 308 CAMAs were characterized via histological and fluorescent in situ hybridization (FISH) approaches respectively, and shown to be distributed extensively throughout coral tissues collected from five sites in Japan and Australia. The densities of CAMAs within the tissues were negatively correlated with the distance from the coastline (i.e. lowest densities at offshore sites). CAMAs were randomly distributed across the six coral tissue regions investigated. Within each CAMA, bacterial cells had similar morphological characteristics, but bacterial morphologies varied among CAMAs, with at least five distinct types identified. Identifying the location of microorganisms associated with the coral host is a prerequisite for understanding their contributions to fitness. Localization of tissue-specific communities housed within CAMAs is particularly important, as these communities are potentially important contributors to vital metabolic functions of the holobiont.

Disease is an emerging threat to coral reef ecosystems worldwide, highlighting the need to understand how environmental conditions interact with coral immune function and associated microbial communities to affect holobiont health. Increased coral disease incidence on reefs adjacent to permanently moored platforms on Australia's Great Barrier Reef provided a unique case study to investigate environment–host–microbe interactions in situ . Here, we evaluate coral-associated bacterial community (16S rRNA amplicon sequencing), immune function (protein-based prophenoloxidase-activating system), and water quality parameters before, during and after a disease event. Over the course of the study, 31% of tagged colonies adjacent to platforms developed signs of white syndrome (WS), while all control colonies on a platform-free reef remained visually healthy. Corals adjacent to platforms experienced significant reductions in coral immune function. Additionally, the corals at platform sites that remained visually healthy throughout the study had reduced bacterial diversity compared to healthy colonies at the platform-free site. Interestingly, prior to the observation of macroscopic disease, corals that would develop WS had reduced bacterial diversity and significantly greater community heterogeneity between colonies compared to healthy corals at the same location. These results suggest that activities associated with offshore marine infrastructure impacts coral immunocompetence and associated bacterial community, which affects the susceptibility of corals to disease.

Environmental data, microbiome composition, water quality, SIMPER, and PERMANOVA results

The concept of symbiosis – defined in 1879 by de Bary as ‘the living together of unlike organisms’ – has a rich and convoluted history in biology. In part, because it questioned the concept of the individual, symbiosis fell largely outside mainstream science and has traditionally received less attention than other research disciplines. This is gradually changing. In nature organisms do not live in isolation but rather interact with, and are impacted by, diverse beings throughout their life histories. Symbiosis is now recognized as a central driver of evolution across the entire tree of life, including, for example, bacterial endosymbionts that provide insects with vital nutrients and the mitochondria that power our own cells. Symbioses between microbes and their multicellular hosts also underpin the ecological success of some of the most productive ecosystems on the planet, including hydrothermal vents and coral reefs. In November 2017, scientists working in fields spanning the life sciences came together at a Company of Biologists’ workshop to discuss the origin, maintenance, and long-term implications of symbiosis from the complementary perspectives of cell biology, ecology, evolution and genomics, taking into account both model and non-model organisms. Here, we provide a brief synthesis of the fruitful discussions that transpired., Summary: At a recent Company of Biologists workshop, evolutionary biologists discussed the major outstanding questions in symbiosis research.

Coral reefs are currently in steep decline worldwide due to changes in climate and anthropogenic activity. Given reefs’ key roles as centers of biodiversity and the variety of services they provide for humans, it is imperative that we develop reef management strategies that are sensitive to environmental changes and that allow timely interventions in response to specific threats. The use of bioindicators has been demonstrated as an effective way to monitor a broad range of ecosystems, and coral microbiomes show immense potential as bioindicators for coral reefs. Given the decline of coral reefs worldwide, and the diversity of species that are currently under threat, coral microbiomes can provide much-needed insights and information for the purposes of reef conservation and protection.

Disease outbreaks are implicated in coral reef degradation worldwide, but little is known about the role of viruses in coral health. In this study, transmission electron microscopy (TEM) was employed in parallel with flow cytometry to compare viral communities associated with visually healthy and white syndrome (WS)-infected tissues of the coral Acropora hyacinthus at Lizard Island on Australia's northern Great Barrier Reef. Viral community shifts were observed on WS-infected corals that were characterized by higher abundance, smaller size and distinct morphology of virus-like particles (VLPs) on disease lesions relative to healthy tissues. Coral tissues displaying WS contained 65% more VLPs, with 87% of these falling in the sub-100 nm size range, compared to only 7% from healthy tissues. While the observed viral community shifts are not necessarily indicative of disease causation, they may provide diagnostic criteria to discriminate between distinct, but macroscopically similar, WS and WS-like coral diseases. Furthermore, these results highlight the need to incorporate virology in investigations of coral health and disease.

Brown band (BrB) disease manifests on corals as a ciliate-dominated lesion that typically progresses rapidly causing extensive mortality, but it is unclear whe- ther the dominant ciliate Porpostoma guamense is a pri- mary or an opportunistic pathogen, the latter taking advantage of compromised coral tissue or depressed host resistance. In this study, manipulative aquarium-based experiments were used to investigate the role of P. gua- mense as a pathogen when inoculated onto fragments of the coral Acropora hyacinthus that were either healthy, preyed on by Acanthaster planci (crown-of-thorns starfish; COTS), or experimentally injured. Following ciliate inoc- ulation, BrB lesions developed on all of COTS-predated fragments (n = 9 fragments) and progressed up to 4.6 ± 0.3 cm d -1 , resulting in *70 % of coral tissue loss after 4 d. Similarly, BrB lesions developed rapidly on experimentally injured corals and *38 % of coral tissue area was lost 60 h after inoculation. In contrast, no BrB lesions were observed on healthy corals following experi- mental inoculations. A choice experiment demonstrated that ciliates are strongly attracted to physically injured corals, with over 55 % of inoculated ciliates migrating to injured corals and forming distinct lesions, whereas ciliates did not migrate to healthy corals. Our results indicate that ciliates characteristic of BrB disease are opportunistic pathogens that rapidly migrate to and colonise compro- mised coral tissue, leading to rapid coral mortality, par- ticularly following predation or injury. Predicted increases in tropical storms, cyclones, and COTS outbreaks are likely to increase the incidence of coral injury in the near future, promoting BrB disease and further contributing to declines in coral cover.

Coral tissue loss diseases, collectively known as white syndromes (WSs), induce significant mortality on reefs throughout the Indo-Pacific, yet definitive confirmation of WS etiologies remains elusive. In this study, we integrated ecological disease monitoring, bacterial community profiling, in situ visualization of microbe-host interactions, and cellular responses of the host coral through an 18-month repeated-sampling regime. We assert that the observed pathogenesis of WS lesions on acroporid corals at Lizard Island (Great Barrier Reef) is not the result of apoptosis or infection by Vibrio bacteria, ciliates, fungi, cyanobacteria, or helminths. Histological analyses detected helminths, ciliates, fungi, and cyanobacteria in fewer than 25% of WS samples, and helminths and fungi were also observed in 12% of visually healthy samples. The abundances of Vibrio -affiliated sequences (assessed using 16S rRNA amplicon sequencing) did not differ significantly between health states and never exceeded 3.3% of reads in any individual sample. In situ visualization detected Vibrio bacteria only in summer WS lesion samples and revealed no signs of these bacteria in winter disease samples (or any healthy tissue samples), despite continued disease progression year round. However, a 4-fold increase in Rhodobacteraceae -affiliated bacterial sequences at WS lesion fronts suggests that this group of bacteria could play a role in WS pathogenesis and/or serve as a diagnostic criterion for disease differentiation. While the causative agent(s) underlying WSs remains elusive, the microbial and cellular processes identified in this study will help to identify and differentiate visually similar but potentially distinct WS etiologies. IMPORTANCE Over the past decade, a virulent group of coral diseases known as white syndromes have impacted coral reefs throughout the Indian and Pacific Oceans. This article provides a detailed case study of white syndromes to combine disease ecology, high-throughput microbial community profiling, and cellular-scale host-microbe visualization over seasonal time scales. We provide novel insights into the etiology of this devastating disease and reveal new diagnostic criteria that could be used to differentiate visually similar but etiologically distinct forms of white syndrome.

In recent decades, coral reef ecosystems have declined to the extent that reefs are now threatened globally. While many water quality parameters have been proposed to contribute to reef declines, little evidence exists conclusively linking specific water quality parameters with increased disease prevalence in situ. Here we report evidence from in situ coral health surveys confirming that chronic exposure to dredging-associated sediment plumes significantly increase the prevalence of white syndromes, a devastating group of globally important coral diseases. Coral health surveys were conducted along a dredging-associated sediment plume gradient to assess the relationship between sedimentation, turbidity and coral health. Reefs exposed to the highest number of days under the sediment plume (296 to 347 days) had two-fold higher levels of disease, largely driven by a 2.5-fold increase in white syndromes, and a six-fold increase in other signs of compromised coral health relative to reefs with little or no plume exposure (0 to 9 days). Multivariate modeling and ordination incorporating sediment exposure level, coral community composition and cover, predation and multiple thermal stress indices provided further confirmation that sediment plume exposure level was the main driver of elevated disease and other compromised coral health indicators. This study provides the first evidence linking dredging-associated sedimentation and turbidity with elevated coral disease prevalence in situ. Our results may help to explain observed increases in global coral disease prevalence in recent decades and suggest that minimizing sedimentation and turbidity associated with coastal development will provide an important management tool for controlling coral disease epizootics.

A real-time quantitative PCR-based detection assay targeting the dnaJ gene (encoding heat shock protein 40) of the coral pathogen Vibrio coralliilyticus was developed. The assay is sensitive, detecting as little as 1 CFU per ml in seawater and 10 4 CFU per cm 2 of coral tissue. Moreover, inhibition by DNA and cells derived from bacteria other than V. coralliilyticus was minimal. This assay represents a novel approach to coral disease diagnosis that will advance the field of coral disease research.

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.

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)

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 20984(T) . 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.

Vibrio coralliilyticus is an important coral pathogen demonstrated to cause disease outbreaks worldwide. This study investigated the feasibility of applying bacteriophage therapy to treat the coral pathogen V. coralliilyticus. A specific bacteriophage for V. coralliilyticus strain P1 (LMG23696), referred to here as bacteriophage YC, was isolated from the seawater above corals at Nelly Bay, Magnetic Island, central Great Barrier Reef (GBR), the same location where the bacterium was first isolated. Bacteriophage YC was shown to be a lytic phage belonging to the Myoviridae family, with a rapid replication rate, high burst size, and high affinity to its host. By infecting its host bacterium, bacteriophage YC was able to prevent bacterial-induced photosystem inhibition in pure cultures of Symbiodinium, the photosymbiont partner of coral and a target for virulence factors produced by the bacterial pathogen. Phage therapy experiments using coral juveniles in microtiter plates as a model system revealed that bacteriophage YC was able to prevent V. coralliilyticus-induced photoinactivation and tissue lysis. These results demonstrate that bacteriophage YC has the potential to treat coral disease outbreaks caused by the bacterial pathogen V. coralliilyticus, making it a good candidate for phage therapy treatment of coral disease.

Coral disease has emerged over recent decades as a significant threat to coral reef ecosystems, with declines in coral cover and diversity of Caribbean reefs providing an example of the potential impacts of disease at regional scales. If similar trends are to be mitigated or avoided on reefs worldwide, a deeper understanding of the factors underlying the origin and spread of coral diseases and the steps that can be taken to prevent, control, or reduce their impacts is required. In recent years, an increased focus on coral microbiology and the application of classic culture techniques and emerging molecular technologies has revealed several coral pathogens that could serve as targets for novel coral disease diagnostic tools. The ability to detect and quantify microbial agents identified as indicators of coral disease will aid in the elucidation of disease causation and facilitate coral disease detection and diagnosis, pathogen monitoring in individuals and ecosystems, and identification of pathogen sources, vectors, and reservoirs. This information will advance the field of coral disease research and contribute knowledge necessary for effective coral reef management. This paper establishes the need for sensitive and specific molecular-based coral pathogen detection, outlines the emerging technologies that could serve as the basis of a new generation of coral disease diagnostic assays, and addresses the unique challenges inherent to the application of these techniques to environmentally derived coral samples.