Identification of putative coral pathogens in endangered Caribbean staghorn coral using machine learning.

Coral diseases contribute to the rapid decline in coral reefs worldwide, and yet coral bacterial pathogens have proved difficult to identify because 16S rRNA gene surveys typically identify tens to hundreds of disease-associate bacteria as putative pathogens. An example is white band disease (WBD), which has killed up to 95% of the now-endangered Caribbean Acropora corals since 1979, yet the pathogen is still unknown. The 16S rRNA gene surveys have identified hundreds of WBD-associated bacterial amplicon sequencing variants (ASVs) from at least nine bacterial families with little consensus across studies. We conducted a multi-year, multi-site 16S rRNA gene sequencing comparison of 269 healthy and 143 WBD-infected Acropora cervicornis and used machine learning modelling to accurately predict disease outcomes and identify the top ASVs contributing to disease. Our ensemble ML models accurately predicted disease with greater than 97% accuracy and identified 19 disease-associated ASVs and five healthy-associated ASVs that were consistently differentially abundant across sampling periods. Using a tank-based transmission experiment, we tested whether the 19 disease-associated ASVs met the assumption of a pathogen and identified two pathogenic candidate ASVs-ASV25 Cysteiniphilum litorale and ASV8 Vibrio sp. to target for future isolation, cultivation, and confirmation of Henle-Koch's postulate via transmission assays.

Sea-Air Transfer of Ostreopsis Phycotoxins Is Driven by the Chemical Diversity of the Particulate Fraction in the Surface Microlayer.

Blooms of Ostreopsis cf. ovata pose an emerging health threat, causing respiratory disorders in various coastal regions. This dinoflagellate produce potent phycotoxins named ovatoxins that can be transferred from the seawater to the atmosphere. However, the biotic and abiotic conditions affecting their transfer are still unknown. In this study, we investigate the sea-to-air transfer of O. cf ovata phycotoxins using a process study in an aerosol reference tank (MART) and field observations. The process study exhibited a positive correlation between the phycotoxin content in sea spray aerosol (up to 832.59 ng m-3) and the particulate phycotoxin fraction in the water column and surface microlayer. In contrast, in the natural system, aerosolized phycotoxins were only observed in one out of six air collection (total toxins 0.59 ng m-3) despite optimal wind conditions. In both the process study and the natural system, ovatoxins represented only a minor fraction of the total toxin content, which was comprised of up to 90% liguriatoxins. In seawater, while no solubilized ovatoxins were detected, the concentration in dissolved liguriatoxin-a reached up to 19.07 µg L-1. These results underscore the need for future research on the liguriatoxins, and on their toxicity to establish safe exposure thresholds for beachgoers.

On the Hunt for New Toxin Families Produced by a Mediterranean Strain of the Benthic Dinoflagellate Ostreopsis cf. ovata.

Ostreopsis cf. ovata is a benthic dinoflagellate known to produce palytoxin (PLTX) and its analogues. Recent investigations suggested the production of unknown toxins by a Mediterranean strain. In the present work, two new families of toxins, potentially novel in their structures, were purified from this same Mediterranean strain of Ostreopsis cf. ovata. The low amount of material isolated only allowed for acquisition of high-resolution mass spectrometry data and the evaluation of their cytotoxicity to human lung cancer cells. Based on their HRMS data, none of these new compounds appear to be close PLTX analogues, although their mass spectra suggest poly-hydroxylated long chain compounds of high molecular weight (1370-2143 Da). The cell cytotoxicity concentrations (CC50) of these new purified toxins ranged between 0.68 and 3.12 µg/mL, and this was enhanced when they were tested as mixtures, suggesting synergistic effects of Ostreopsis toxins. The two families of compounds were named the liguriatoxins (LGTX) and rivieratoxins (RVTX), with each family containing three members. Additional work on purification is needed to fully characterize the structures of these six new dinoflagellate toxins.