Sea urchin larvae decipher the epiphytic bacterial community composition when selecting sites for attachment and metamorphosis.

Most marine invertebrates have dispersive larvae and relatively immobile adults. These developmental stages are linked by a settlement event, which is often mediated by specific cues in bacterial biofilms. While larvae distinguish between biofilms from different environments, it remains unknown if they receive information from all, only a few or even just a single bacterial species in natural biofilms. Here we asked how specific is larval settlement to the bacterial community structure and/or taxonomically distinguishable groups of bacteria in epiphytic marine biofilms? We used novel multivariate statistical approaches to investigate if larval settlement of two sea urchins correlated with the microbial community composition. Larval settlement of Heliocidaris erythrogramma revealed a strong correlation with the community composition, highlighted by canonical analysis of principle components, a constrained ordination technique. Using this technique, the importance of operational taxonomic units (OTUs) within communities relative to larval settlement was investigated. Larval settlement not only correlated, both positively and negatively, with the epiphytic bacterial community composition but also with the relative abundance of few OTUs within these communities. In contrast, no such correlation was observed for the other urchin, Holopneustes purpurascens, whose larvae likely respond to bacterial biofilms in a more general way and specifically respond to a defined settlement cue of algal origin.

Do cues matter? Highly inductive settlement cues don't ensure high post-settlement survival in sea urchin aquaculture.

Increasing settlement and post-settlement survival during the critical transition from planktonic larvae to benthic juveniles will increase efficiency for sea urchin aquaculture. This study investigated the effects of temperature and settlement cues on the settlement and post-settlement survival of the sea urchin Tripneustes gratilla during this phase. The current commercial methodology, which utilises natural biofilm settlement plates, was tested and resulted in low settlement (<2%) and poor post-settlement survival (<1% of settled urchins). In laboratory trials, settlement was high and unaffected by temperatures between 24 and 30°C, but significantly decreased at 33°C. Development of spines, however, was significantly affected by temperatures over 29°C. Mirroring this result, post-settlement survival was optimal between 24-28°C. In laboratory assays, the macroalgae Sargassum linearifolium and Corallina officinalis, and seawater conditioned with these algae, induced significantly higher settlement (>90%) than a natural biofilm (∼25%). The addition of macroalgae-conditioned seawater to natural biofilm significantly increased settlement rates (>85%). Mixed consortia and single strains of bacteria isolated from macroalgae, biofilms and adult conspecifics all induced significant settlement, but at significantly lower rates than macroalgae. No evidence was found that higher rates of settlement to bacteria on macroalgae were generated by a cofactor from the macroalgae. Age of bacterial cultures, culturing bacteria on solid and liquid media and concentration of nutrients in cultures had little effect on settlement rates. Finally, macroalgae-conditioned seawater combined with natural biofilm settlement plates induced significantly higher settlement than to the biofilm plates alone in a commercial scale trial. However, high post-settlement mortality resulted in equivalent survival between treatments after 25 days. This study highlights that settlement studies should extend to post-settlement survival, which remains poor for T. gratilla and is a significant obstacle to increasing efficiency for aquaculture.