Recruitment via settlement of pelagic larvae is critical for the persistence of benthic marine populations. For many benthic invertebrates, larval settlement occurs in response to surface microbial films. Larvae of the serpulid polychaete Hydroides elegans can be induced to settle by single bacterial species. Until now, only Pseudoalteromonas luteoviolacea had been subjected to detailed genetic and mechanistic studies. To determine if the complex structures, termed tailocins, derived from phage-tail gene assemblies and hypothesized to be the settlement cue in P. luteoviolacea were present in all inductive bacteria, genomic comparisons with inductive strains of Cellulophaga lytica, Bacillus aquimaris and Staphylococcus warneri were undertaken. They revealed that the gene assemblies for tailocins are lacking in these other bacteria. Negatively stained TEM images confirmed the absence of tailocins and revealed instead large numbers of extracellular vesicles in settlement-inductive fractions from all three bacteria. TEM imaging confirmed for C. lytica that the vesicles are budded from cell surfaces in a manner consistent with the production of outer membrane vesicles. Finding multiple bacteria settlement cues highlights the importance of further studies into the role of bacterial extracellular vesicles in eliciting settlement and metamorphosis of benthic marine larvae.

The microbial community composition in affected and unaffected portions of diseased sponges and healthy control sponges of Aplysina aerophoba was assessed to ascertain the role of microbes in the disease process. Sponge secondary metabolites were also examined to assess chemical shifts in response to infection. The microbial profile and aplysinimine levels in unaffected tissue near the lesions closely reflected those of healthy sponge tissue, indicating a highly localized disease process. DGGE detected multiple sequences that were exclusively present in diseased sponges. Most notably, a Deltaproteobacteria sequence with high homology to a coral black band disease strain was detected in all sponge lesions and was absent from all healthy and unaffected regions of diseased sponges. Other potential pathogens identified by DGGE include an environmental Cytophaga strain and a novel Epsilonproteobacteria strain with no known close relatives. The disease process also caused a major shift in prokaryote community structure at a very high taxonomic level. Using 16S rRNA gene sequence analysis, only the diseased sponges were found to contain sequences belonging to the Epsilonproteobacteria and Firmicutes, and there was a much greater number of Bacteroidetes sequences within the diseased sponges. In contrast, only the healthy sponges contained sequences corresponding to the cyanobacteria and 'OP1' candidate division, and the healthy sponges were dominated by Chloroflexi and Gammaproteobacteria sequences. Overall bacterial diversity was found to be considerably higher in diseased sponges than in healthy sponges. These results provide a platform for future cultivation-based experiments to isolate the putative pathogens from A. aerophoba and perform re-infection trials to define the disease aetiology.

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Dereplication of a methanolic extract of the marine sponge Ianthella flabelliformis using FTICR-MS accurate mass determination and MS(n) techniques enabled rapid and unambiguous detection of a new compound among a plethora of known compounds. Isolation of this compound and the known 19-deoxy analogue using the hyphenated technique LC-UV/MS-SPE-NMR was undertaken, and the structures were confirmed, from a single chromatographic run, as 19-hydroxyaraplysillin-I N(20)-sulfamate (1) and araplysillin-I N(20)-sulfamate (2).

Significance New surfaces in the sea are quickly populated by dense communities of invertebrate animals, whose establishment and maintenance require site-specific settlement of larvae from the plankton. Larvae selectively settle in sites where they can metamorphose and thrive largely due to inductive cues from bacteria residing on these surfaces. However, the nature of the cues used to identify “right places” has remained enigmatic. Here, we demonstrate that lipopolysaccharide, the main component of the outer membrane of Gram-negative bacteria from the bacterium Cellulophaga lytica , induces metamorphosis for a marine worm. We then discuss the likelihood that lipopolysaccharide provides the variation necessary to explain settlement site selectivity for many of the bottom-living invertebrate animals that metamorphose in response to bacterial biofilms.