Marine Envenomation in Okinawa: Overview and Treatment Concept.

Okinawa prefecture is a popular tourist destination due to its beaches and reefs. The reefs host a large variety of animals, including a number of venomous species. Because of the popularity of the reefs and marine activities, people are frequently in close contact with dangerous venomous species and, thus, are exposed to potential envenomation. Commonly encountered venomous animals throughout Okinawa include the invertebrate cone snail, sea urchin, crown-of-thorns starfish, blue-ringed octopus, box jellyfish, and fire coral. The vertebrates include the stonefish, lionfish, sea snake, and moray eel. Treatment for marine envenomation can involve first aid, hot water immersion, antivenom, supportive care, regional anesthesia, and pharmaceutical administration. Information on venomous animals, their toxins, and treatment should be well understood by prehospital care providers and physicians practicing in the prefecture.

Captivity induces a sweeping and sustained genomic response in a starfish.

Marine animals in the wild are often difficult to access, so they are studied in captivity. However, the implicit assumption that physiological processes of animals in artificial environments are not different from those in the wild has rarely been tested. Here, we investigate the extent to which an animal is impacted by captivity by comparing global gene expression in wild and captive crown-of-thorns starfish (COTS). In a preliminary analysis, we compared transcriptomes of three external tissues obtained from multiple wild COTS with a single captive COTS maintained in aquaria for at least 1 week. On average, an astonishingly large 24% of the coding sequences in the genome were differentially expressed. This led us to conduct a replicated experiment to test more comprehensively the impact of captivity on gene expression. Specifically, a comparison of 13 wild with 8 captive COTS coelomocyte transcriptomes revealed significant differences in the expression of 20% of coding sequences. Coelomocyte transcriptomes in captive COTS remain different from those in wild COTS for more than 30 days and show no indication of reverting back to a wild state (i.e. no evidence of acclimation). Genes upregulated in captivity include those involved in oxidative stress and energy metabolism, whereas genes downregulated are involved in cell signalling. These changes in gene expression indicate that being translocated and maintained in captivity has a marked impact on the physiology and health of these echinoderms. This study suggests that caution should be exercised when extrapolating results from captive aquatic invertebrates to their wild counterparts.

Improving coral cover using an integrated pest management framework.

Integrated pest management (IPM) leverages our understanding of ecological interactions to mitigate the impact of pest species on economically and/or ecologically important assets. It has primarily been applied in terrestrial settings (e.g., agriculture), but has rarely been attempted for marine ecosystems. The crown-of-thorns starfish (CoTS), Acanthaster spp., is a voracious coral predator throughout the Indo-Pacific where it undergoes large population increases (irruptions), termed outbreaks. During outbreaks CoTS act as a pest species and can result in substantial coral loss. Contemporary management of CoTS on the Great Barrier Reef (GBR) adopts facets of the IPM paradigm to manage these outbreaks through strategic use of direct manual control (culling) of individuals in response to ecologically based target thresholds. There has, however, been limited quantitative analysis of how to optimize the implementation of such thresholds. Here we use a multispecies modeling approach to assess the performance of alternative CoTS management scenarios for improving coral cover trajectories. The scenarios examined varied in terms of their ecological threshold target, the sensitivity of the threshold, and the level of management resourcing. Our approach illustrates how to quantify multidimensional trade-offs in resourcing constraints, concurrent CoTS and coral population dynamics, the stringency of target thresholds, and the geographical scale of management outcomes (number of sites). We found strategies with low target density thresholds for CoTS (≤0.03 CoTS min-1 ) could act as "Effort Sinks" and limit the number of sites that could be effectively controlled, particularly under CoTS population outbreaks. This was because a handful of sites took longer to control, which meant other sites were not controlled. Higher density thresholds (e.g., 0.04-0.08 CoTS min-1 ), tuned to levels of coral cover, diluted resources among sites but were more robust to resourcing constraints and pest population dynamics. Our study highlights trade-off decisions when using an IPM framework and informs the implementation of threshold-based strategies on the GBR.

Phytoplankton community structure and environmental factors during the outbreak of Crown-of-Thorns Starfish in Xisha Islands, South China Sea.

The "larval starvation hypothesis" proposed that the growing frequency of Crown-of-Thorns Starfish (CoTS) outbreaks could be attributed to increased availability of phytoplankton. However, comprehensive field investigation on the living environment of CoTS larvae and the availability of phytoplankton are still lacking. A cruise was conducted in June 2022 in Xisha Islands, South China Sea, to study the interaction between environmental conditions and phytoplankton communities during CoTS outbreak period. The average concentrations of dissolved inorganic phosphorus (0.05 ± 0.01 µmol L-1), dissolved inorganic nitrogen (0.66 ± 0.8 µmol L-1) and chlorophyll a (0.05 ± 0.05 µg L-1) suggested that phytoplankton may be limited for CoTS larvae in Xisha Islands. Microscopic observation and high-throughput sequencing were used to study the composition and structure of the phytoplankton communities. Bacillariophyta predominated in phytoplankton communities with the highest abundance and species richness. 29 dominant species, including 4 species with size-range preferred by CoTS larvae, were identified in Xisha Islands. The diversity index of all stations indicated a species-rich and structure-stable phytoplankton community in Xisha Islands during the period of CoTS outbreak, which may contribute to CoTS outbreak. These findings revealed the structure of phytoplankton community and environmental factors in the study area during CoTS outbreak, providing the groundwork for future research into the causes and processes of CoTS outbreak.

Sex-specific expression of pheromones and other signals in gravid starfish.

BACKGROUND: Many echinoderms form seasonal aggregations prior to spawning. In some fecund species, a spawning event can lead to population outbreaks with detrimental ecosystem impacts. For instance, outbreaks of crown-of-thorns starfish (COTS), a corallivore, can destroy coral reefs. Here, we examine the gene expression in gravid male and female COTS prior to spawning in the wild, to identify genome-encoded factors that may regulate aggregation and spawning. This study is informed by a previously identified exoproteome that attracts conspecifics. To capture the natural gene expression profiles, we isolated RNAs from gravid female and male COTS immediately after they were removed from the Great Barrier Reef.  RESULTS: Sexually dimorphic gene expression is present in all seven somatic tissues and organs that we surveyed and in the gonads. Approximately 40% of the exoproteome transcripts are differentially expressed between sexes. Males uniquely upregulate an additional 68 secreted factors in their testes. A suite of neuropeptides in sensory organs, coelomocytes and gonads is differentially expressed between sexes, including the relaxin-like gonad-stimulating peptide and gonadotropin-releasing hormones. Female sensory tentacles-chemosensory organs at the distal tips of the starfish arms-uniquely upregulate diverse receptors and signalling molecules, including chemosensory G-protein-coupled receptors and several neuropeptides, including kisspeptin, SALMFamide and orexin. CONCLUSIONS: Analysis of 103 tissue/organ transcriptomes from 13 wild COTS has revealed genes that are consistently differentially expressed between gravid females and males and that all tissues surveyed are sexually dimorphic at the molecular level. This finding is consistent with female and male COTS using sex-specific pheromones to regulate reproductive aggregations and synchronised spawning events. These pheromones appear to be received primarily by the sensory tentacles, which express a range of receptors and signalling molecules in a sex-specific manner. Furthermore, coelomocytes and gonads differentially express signalling and regulatory factors that control gametogenesis and spawning in other echinoderms.

Characterization and localization of relaxin-like gonad-stimulating peptide in the crown-of-thorns starfish, Acanthaster cf. solaris.

In starfish, a relaxin-like gonad-stimulating peptide (RGP) is the gonadotropin responsible for final gamete maturation. RGP comprises two different peptides, A- and B-chains with two interchain and one intrachain disulfide bonds. The existence of two isomers of RGP in the crown-of-thorns starfish, Acanthaster planci, has been reported previously, but it was recently shown that A. planci represents a species complex with four different species. Here we elucidated the authentic sequence of the Pacific species, Acanthaster cf. solaris, RGP (Aso-RGP). The Aso-RGP precursor encoded by a 354 base pair open reading frame was composed of 117 amino acids (aa). The amino acid identity of Aso-RGP to Patiria pectinifera RGP (Ppe-RGP) and Asterias amurensis RGP (Aam-RGP) was 74% and 60%, respectively. Synthetic Aso-RGP induced spawning of ovarian fragments from A. cf. solaris. Ppe-RGP and Aam-RGP also induced spawning by A. cf. solaris ovaries. In contrast, Ppe-RGP and Aso-RGP induced spawning by P. pectinifera ovaries, but Aam-RGP was inactive. Notably, anti-Ppe-RGP antibodies recognized Aso-RGP as well as Ppe-RGP. Localization of Aso-RGP was observed immunohistochemically using anti-Ppe-RGP antibodies, showing that Aso-RGP was mainly present in the radial nerve cords of A. cf. solaris. Aso-RGP was distributed not only in the epithelium of the ectoneural region but also in the neuropile of the ectoneural region. These results suggest that Aso-RGP is synthesized in the epithelium of the ectoneural region, then transferred to fibers in the neuropile of the ectoneural region in radial nerve cords.

The hidden army: corallivorous crown-of-thorns seastars can spend years as herbivorous juveniles.

Crown-of-thorns seastar (COTS) outbreaks are a major threat to coral reefs. Although the herbivorous juveniles and their switch to corallivory are key to seeding outbreaks, they remain a black box in our understanding of COTS. We investigated the impact of a delay in diet transition due to coral scarcity in cohorts reared on crustose coralline algae for 10 months and 6.5 years before being offered coral. Both cohorts achieved an asymptotic size (16-18 mm diameter) on algae and had similar exponential growth on coral. After 6.5 years of herbivory, COTS were competent coral predators. This trophic and growth plasticity results in a marked age-size disconnect adding unappreciated complexity to COTS boom-bust dynamics. The potential that herbivorous juveniles accumulate in the reef infrastructure to seed outbreaks when favourable conditions arise has implications for management of COTS populations.

High-resolution characterization of the abiotic environment and disturbance regimes on the Great Barrier Reef, 1985-2017.

This data compilation synthesizes 36 static environmental and spatial variables, and temporally explicit modeled estimates of three major disturbances to coral cover on the Great Barrier Reef (GBR): (1) coral bleaching, (2) tropical cyclones, and (3) outbreaks of the coral-eating crown-of-thorns starfish Acanthaster cf. solaris. Data are provided on a standardized grid (0.01° × 0.01° ~ 1 × 1 km) for reef locations along the GBR, containing 15,928 pixels and excluding the northernmost sections (<12° S) where empirical data were sparse. This compilation provides a consistent and high-resolution characterization of the abiotic environment and disturbance regimes for GBR reef locations at a fine spatial scale to be used in the development of complex ecosystem models. Static estimates of environmental variables (e.g., depth, bed shear stress, average temperature, temperature variation) originally developed by the Commonwealth of Australia's Environment Research Facility (CERF) Marine Biodiversity Hub were provided by Geoscience Australia. Annual (1985-2017) disturbance estimates were either interpolated from empirical data (A. cf. solaris), predicted from proxy indicators (e.g., degree heating weeks [DHW] as a proxy for bleaching severity), or explicitly modeled (e.g., wave height model for each cyclone). This data set synthesizes some of the most recent advances in remote sensing and modeling of environmental conditions on the GBR; yet it is not exhaustive and we highlight areas that should be expanded through future research. The characterization of abiotic and disturbance regimes presented here represent an essential tool for the development of complex regional scale models of the GBR; preventing redundancy between working groups and promoting collaboration, innovation, and consistency. When using the data set, we kindly request that you cite this article and/or the articles cited in the reference section, recognizing the work that went into compiling the data together and the original authors' willingness to make it publicly available.

Great Barrier Reef recovery through multiple interventions.

The decline of coral cover on Australia's Great Barrier Reef (GBR) has largely been attributed to the cumulative pressures of tropical cyclones, temperature-induced coral bleaching, and predation by crown-of-thorns starfish (CoTS). In such a complex system, the effectiveness of any management intervention will become apparent only over decadal time scales. Systems modeling approaches are therefore essential to formulating and testing alternative management strategies. For a network of reefs, we developed a metacommunity model that incorporated the cumulative pressures of tropical cyclones, coral bleaching, predation, and competition between corals. We then tested the response of coral cover to management interventions including catchment restoration to reduce discharge onto the reef during cyclone-induced flood events and enhanced protection of trophic networks supporting predation of CoTS. Model results showed good agreement with long-term monitoring of the GBR, including cyclical outbreaks of CoTS driven by predator-prey dynamics on the network of reefs. Testing of intervention strategies showed that catchment restoration would likely improve coral cover. However, strategies that combined catchment restoration with enhanced CoTS predation were far more effective than catchment restoration alone.

Chemical Ecology of Chemosensation in Asteroidea: Insights Towards Management Strategies of Pest Species.

Within the Phylum Echinodermata, the class Asteroidea, commonly known as starfish and sea stars, encompasses a large number of benthos inhabiting genera and species with various feeding modalities including herbivores, carnivores, omnivores and detritivores. The Asteroidea rely on chemosensation throughout their life histories including hunting prey, avoiding or deterring predators, in the formation of spawning aggregations, synchronizing gamete release and targeting appropriate locations for larval settlement. The identities of many of the chemical stimuli that mediate these physiological and behavioural processes remain unresolved even though evidence indicates they play pivotal roles in the functionality of benthic communities. Aspects of chemosensation, as well as putative chemically-mediated behaviours and the molecular mechanisms of chemoreception, within the Asteroidea are reviewed here, with particular reference to the coral reef pest the Crown-of-Thorns starfish Acanthaster planci species complex, in the context of mitigation of population outbreaks.

Environmental and Ecological Effects of Climate Change on Venomous Marine and Amphibious Species in the Wilderness.

INTRODUCTION: Recent analyses of data show a warming trend in global average air and sea surface ocean temperatures. The atmosphere and ocean have warmed, the amounts of snow and ice have diminished, the sea level has risen, and the concentrations of greenhouse gases have increased. This article will focus on climate change and projected effects on venomous marine and amphibious creatures with the potential impact on human health. METHODS: Retrospective analysis of environmental, ecological, and medical literature with a focus on climate change, toxinology, and future modeling specific to venomous aquatic and amphibious creatures. Species included venomous jellyfish, poisonous fish, crown-of-thorns starfish, sea snakes, and toxic frogs. RESULTS: In several projected scenarios, rising temperatures, weather extremes, and shifts in seasons will increase poisonous population numbers, particularly with certain marine creatures like jellyfish and crown-of-thorns starfish. Habitat expansions by lionfish and sea snakes are projected to occur. These phenomena, along with increases in human populations and coastal development will likely increase human-animal encounters. Other species, particularly amphibious toxic frogs, are declining rapidly due to their sensitivity to any temperature change or subtle alterations in the stability of their environment. If temperatures continue to rise to record levels over the next decades, it is predicted that the populations of these once plentiful and critically important animals to the aquatic ecosystem will decline and their geographic distributions will shrink. CONCLUSION: Review of the literature investigating the effect and forecasts of climate change on venomous marine and amphibious creatures has demonstrated that temperature extremes and changes to climatic norms will likely have a dramatic effect on these toxicological organisms. The effects of climate change on these species through temperature alteration and rising coastal waters will influence each species differently and in turn potentially affect commercial industries, travel, tourism, and human health.

Indirect effects of ocean acidification drive feeding and growth of juvenile crown-of-thorns starfish, Acanthaster planci.

The indirect effects of changing climate in modulating trophic interactions can be as important as the direct effects of climate stressors on consumers. The success of the herbivorous juvenile stage of the crown-of-thorns starfish (COTS), Acanthaster planci, may be affected by the impacts of ocean conditions on its crustose coralline algal (CCA) food. To partition the direct effects of near future ocean acidification on juvenile COTS and indirect effects through changes in their CCA food, COTS were grown in three pHT levels (7.9, 7.8, 7.6) and fed CCA grown at similar pH levels. Consumption of CCA by COTS was bolstered when the COTS were grown in low pH and when they were fed CCA grown in low pH regardless of the pH in which the COTS were reared. COTS fed CCA grown at pH 7.6 grew fastest, but the pH/pCO2 that the COTS were reared in had no direct effect on growth. Ocean acidification conditions decreased the C : N ratio and carbonate levels in the CCA. Bolstered growth in COTS may be driven by enhanced palatability, increased nutritive state and reduced defences of their CCA food. These results indicate that near future acidification will increase the success of early juvenile COTS and boost recruitment into the coral-eating life stage.

Marine protected areas increase resilience among coral reef communities.

With marine biodiversity declining globally at accelerating rates, maximising the effectiveness of conservation has become a key goal for local, national and international regulators. Marine protected areas (MPAs) have been widely advocated for conserving and managing marine biodiversity yet, despite extensive research, their benefits for conserving non-target species and wider ecosystem functions remain unclear. Here, we demonstrate that MPAs can increase the resilience of coral reef communities to natural disturbances, including coral bleaching, coral diseases, Acanthaster planci outbreaks and storms. Using a 20-year time series from Australia's Great Barrier Reef, we show that within MPAs, (1) reef community composition was 21-38% more stable; (2) the magnitude of disturbance impacts was 30% lower and (3) subsequent recovery was 20% faster that in adjacent unprotected habitats. Our results demonstrate that MPAs can increase the resilience of marine communities to natural disturbance possibly through herbivory, trophic cascades and portfolio effects.

Plancitoxin I from the venom of crown-of-thorns starfish (Acanthaster planci) induces oxidative and endoplasmic reticulum stress associated cytotoxicity in A375.S2 cells.

The crown-of-thorns starfish Acanthaster planci is a venomous starfish whose venom provokes strong cytotoxicity. In the present study, the purified cytotoxic toxin of A. planci venom (CAV) was identified as plancitoxin I protein by mass spectrum analyses. This study aims to investigate the molecular mechanism underlying the cytotoxicity function of plancitoxin I by focusing on the oxidative stress, mitochondrial dysfunction and endoplasmic reticulum (ER) stress pathway in human melanoma A375.S2 cells. The results indicated that after being treated with CAV toxin, A375.S2 cells significantly decreased viability in a dose-dependent manner. The CAV was found to reduce the cellular antioxidant enzymes such as SOD and CAT, and there was a significant decrease in total thiol level and mtDNA integrity, and it enhanced the lipid peroxidation. In addition, CAV increased cytosolic Ca(2+) concentration, and enhanced the expression of the ER molecular chaperones GRP78 and CHOP in a dose-dependent manner. CAV significantly elevated the activity of caspase-3, -8 and -9, and reduced the ratio of Bcl-2/Bax. The cells exhibited apoptosis were determined by using propidium iodide (PI) staining of DNA fragmentation (sub-G1 peak). In summary, the results demonstrated that plancitoxin I inhibits the proliferation of A375.S2 cells through induction of oxidative stress, mitochondrial dysfunction and ER stress associated apoptosis.

Cytotoxic and apoptotic activities of the plancitoxin I from the venom of crown-of-thorns starfish (Acanthaster planci) on A375.S2 cells.

This study reports on a cytotoxic toxin derived from the venom of the crown-of-thorns starfish Acanthaster planci (CAV). The protein toxin was isolated through both ion-exchange and gel-filtration chromatography, and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrum analyzes. The CAV was identified as plancitoxin I protein. The mechanistic role of the CAV toxin was explored in human malignant melanoma A375.S2 cell death. The results indicated that after incubation with CAV toxin, cells significantly decreased in A375.S2 cell viability and increased in the lactate dehydrogenase (LDH) level in a dose-dependent manner. The assays indicated that CAV toxin promoted reactive oxygen species (ROS) production, induced nitric oxide (NO) formation, lost mitochondrial membrane potential (ΔΨm) and induced inter-nucleosomal DNA fragmentation in A375.S2 cells. The molecular cytotoxicity of the CAV toxin was tested through evaluation of the apoptosis/necrosis ratio by double staining with annexin V-FITC and a propidium iodide (PI) assay. The results suggested that CAV toxin induced a cytotoxic effect in A375.S2 cells via the apoptotic procedure, and may be associated with the regulation of the p38 pathways.

A new strategy based on a cascade amplification strategy biosensor for on-site eDNA detection and outbreak warning of crown-of-thorns starfish.

Population outbreaks of the crown-of-thorns starfish (COTS) seriously threaten the sustainability of coral reef ecosystems. However, traditional ecological monitoring techniques cannot provide early warning before the outbreaks, thus preventing timely intervention. Therefore, there is an urgent need for a more accurate and faster technology to predict the outbreaks of COTS. In this work, we developed an electrochemical biosensor based on a programmed catalytic hairpin assembly (CHA) and hybridization chain reaction (HCR) cyclic amplification strategy for sensitive and selective detection of COTS environmental DNA (eDNA) in water bodies. This biosensor exhibited excellent electrochemical characteristics, including a low limit of detection (LOD = 18.4 fM), low limit of quantification (LOQ = 41.1 fM), and wide linear range (50 fM - 10 nM). The biosensing technology successfully allowed the detection of COTS eDNA in the aquarium environment, and the results also demonstrated a significant correlation between eDNA concentration and COTS number (r = 0.990; P < 0.001). The reliability and accuracy of the biosensor results have been further validated through comparison with digital droplet PCR (ddPCR). Moreover, the applicability and accuracy of the biosensor were reconfirmed in field tests at the COTS outbreak site in the South China Sea, which has shown potential application in dynamically monitoring the larvae before the COTS outbreak. Therefore, this efficient electrochemical biosensing technology offers a new solution for on-site monitoring and early warning of the COTS outbreak.

THREE-DECADE changes of reef cover in Pulau Layang-Layang, Malaysia using multitemporal Landsat images.

Over the years, coral reefs in the South China Sea have degraded and faced severe threats from rapid development, coral bleaching, and Crown-of-Thorns Starfish (COTS) outbreak. There is limited knowledge relating to the effects of anthropogenic disturbances and natural events on the coral reefs of Pulau Layang-Layang. This study aims to assess reef cover changes by utilizing Landsat satellite images spanning from 1989 to 2022. Using the object-based image analysis method, this study classified the reef cover into three categories: coral, rock and rubble, and sand. The supervised classification had an overall accuracy of 86.41-87.38 % and Tau's coefficients of 0.80-0.81. The results showed island development and construction of artificial bird sanctuary have led to an increase in coral cover. Furthermore, it was illustrated that the impact of COTS outbreaks in 2010 and 2020 differed significantly, with the latter showing no signs of recovery. Our study underscores the importance of timely intervention to mitigate the spread of COTS. This study provides insights into the resilience and vulnerability of these ecosystems in the face of various stressors.

Crown-of-thorns starfish spines secrete defence proteins.

Background: The crown-of-thorns starfish (COTS; Acanthaster species) is a slow-moving corallivore protected by an extensive array of long, sharp toxic spines. Envenomation can result in nausea, numbness, vomiting, joint aches and sometimes paralysis. Small molecule saponins and the plancitoxin proteins have been implicated in COTS toxicity. Methods: Brine shrimp lethality assays were used to confirm the secretion of spine toxin biomolecules. Histological analysis, followed by spine-derived proteomics helped to explain the source and identity of proteins, while quantitative RNA-sequencing and phylogeny confirmed target gene expression and relative conservation, respectively. Results: We demonstrate the lethality of COTS spine secreted biomolecules on brine shrimp, including significant toxicity using aboral spine semi-purifications of >10 kDa (p > 0.05, 9.82 µg/ml), supporting the presence of secreted proteins as toxins. Ultrastructure observations of the COTS aboral spine showed the presence of pores that could facilitate the distribution of secreted proteins. Subsequent purification and mass spectrometry analysis of spine-derived proteins identified numerous secretory proteins, including plancitoxins, as well as those with relatively high gene expression in spines, including phospholipase A2, protease inhibitor 16-like protein, ependymin-related proteins and those uncharacterized. Some secretory proteins (e.g., vitellogenin and deleted in malignant brain tumor protein 1) were not highly expressed in spine tissue, yet the spine may serve as a storage or release site. This study contributes to our understanding of the COTS through functional, ultrastructural and proteomic analysis of aboral spines.

Ultrasensitive and on-site eDNA detection for the monitoring of crown-of-thorns starfish densities at the pre-outbreak stage using an electrochemical biosensor.

The coral reef crisis has significantly intensified over the last decades, mainly due to severe outbreaks of crown-of-thorns starfish (COTS). Current ecological monitoring has failed to detect COTS densities at the pre-outbreak stage, thus preventing early intervention. In this work, we developed an effective electrochemical biosensor modified by a MoO2/C nanomaterial, as well as a specific DNA probe that could detect trace COTS environmental DNA (eDNA) at a lower detection limit (LOD = 0.147 ng/µL) with excellent specificity. The reliability and accuracy of the biosensor were validated against the standard methods by an ultramicro spectrophotometer and droplet digital PCR (p > 0.05). The biosensor was then utilized for the on-site analysis of seawater samples from SYM-LD and SY sites in the South China Sea. For the SYM-LD site suffering an outbreak, the COTS eDNA concentrations were 0.33 ng/µL (1 m, depth) and 0.26 ng/µL (10 m, depth), respectively. According to the ecological survey, the COTS density was 500 ind/hm2 at the SYM-LD site, verifying the accuracy of our measurements. At the SY site, COTS eDNA was also detected at 0.19 ng/µL, but COTS was not found by the traditional survey. Hence, larvae were possibly present in this region. Therefore, this electrochemical biosensor could be used to monitor COTS populations at the pre-outbreak stages, and potentially serve as a revolutionary early warning method. We will continue to improve this method for picomolar or even femtomolar detection of COTS eDNA.

Significant Changes in Bacterial Communities Associated with Pocillopora Corals Ingestion by Crown-of-Thorns Starfish: An Important Factor Affecting the Coral's Health.

Coral ingestion by crown-of-thorns starfish (COTS) is an important cause of coral reef degradation, although the impacts of COTS feeding on coral-associated microbial communities are not well understood. Therefore, in this study, we analyzed the coral tissue-weight, Symbiodiniaceae density (SD), bacterial community composition, and the predicted functions of bacterial genes associated with Pocillopora corals in healthy portions and feeding scars, following COTS feeding. Coral tissue-weight loss rate in the feeding scars was 71.3-94.95%. The SDs were significantly lower in the feeding scars, and the SD-loss rate was 92.05% ± 2.12%. The relative abundances of bacterial communities associated with Pocillopora corals after COTS feeding changed significantly and were almost completely reorganized at the phylum and genus levels. Analysis of the microbial metagenomic-functional capacities showed that numerous physiological functions of the coral-bacterial holobionts in the feeding scars were different, including amino acid metabolism, xenobiotic biodegradation and metabolism, lipid metabolism, membrane transport, signal transduction, and cell motility, and all these capacities could be corroborated based on metagenomic, transcriptomic or proteomic technologies. Overall, our research suggests that coral holobionts may be destroyed by COTS, and our findings imply that bacterial communities in feeding scars could affect the health of Pocillopora corals.