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A ciliate belonging to theDiadema antillarumscuticociliatosis (DaSc)-associatedPhilasterclade (DaScPc) caused catastrophic long-spined urchin mass mortality in spring and summer of 2022. The ciliate can be grown in culture in both the presence and absence ofD. antillarumtissues, suggesting that it may persist outside its host by consuming microorganisms or dissolved organic nutrients. We hypothesized that DaScPc was present outside its host during and after mass mortality and absent prior to 2022. We examined DaScPc in DNA extracted from 500 swabs of sympatric metazoa and abiotic surfaces, and plankton samples, collected at 35 sites in the Caribbean in 2022 and 2023. DaScPc was detected on corals, turf algae, and a sponge, predominantly at sites with active or prior DaSc. We examined whether it was present prior to 2022 by surveying extracted DNA from Caribbean corals and water collected near corals by PCR and by mining publicly available transcriptomes and metagenomes for DaScPc rRNAs. These efforts yielded no DaScPc genes. We further hypothesized that DaScPc may recruit to the specific corals detected in field surveys, and that these may then infect naïve hosts. A mesocosm experiment to test DaScPc recruitment suggested that, while it recruited to corals, it did so inconsistently between coral species. Incubation of corals that recruited DaScPc with naïve urchins yielded inconclusive results since urchins died without characteristic DaSc signs. Overall, our results suggest that DaScPc may occur outside its urchin host, and that it may have been absent in the region prior to 2022. 

Stony corals (Scleractinia) are invertebrates that form symbiotic relationships with eukaryotic algal endosymbionts and the prokaryotic microbiome. The microbiome has the potential to produce bioactive natural products providing defense and resilience to the coral host against pathogenic microorganisms, but this potential has not been extensively explored. Bacterial pathogens can pose a significant threat to corals, with some species implicated in primary and opportunistic infections of various corals. In response, probiotics have been proposed as a potential strategy to protect corals in the face of increased incidence of disease outbreaks. In this study, we screened bacterial isolates from healthy and diseased corals for antibacterial activity. The bioactive extracts were analyzed using untargeted metabolomics. Herein, UpSet plot and hierarchical clustering analyses were performed to identify isolates with the largest number of unique metabolites. These isolates also displayed different antibacterial activities. Through application of in silico and experimental approaches coupled with genome analysis, we dereplicated natural products from these coral-derived bacteria from Florida’s coral reef environments. The metabolomics approach highlighted in this study serves as a useful resource to select probiotic candidates and enables insights into natural product-mediated chemical ecology in holobiont symbiosis.