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Palytoxin (PTX), one of the most potent and chemically complex marine toxins, is predominantly found in zoanthid corals and sporadically in dinoflagellates. Its biosynthesis and metabolic pathways are largely unknown. However, the widespread occurrence of the toxin in phylogenetically distinct marine organisms is consistent with its production by microorganisms and subsequent accumulation in the food chain. To investigate a possible microbial origin, bacteria from two zoanthid corals (Palythoa caribaeorum, Zoanthus pulchellus) and one sponge (Neofibularia nolitangere) were isolated. More than 250 bacteria were screened for hemolysis using a newly developed PTX-screening assay of which 7% showed PTX-like hemolytic activity. 16S rRNA gene sequencing revealed that these bacterial isolates belonged to strains of Bacillus cereus group (n = 11) as well as the genera Brevibacterium (n = 4) and Acinetobacter (n = 2). The results indicate the presence of Na+/K+-ATPase toxins and possibly PTX in hemolytic bacteria from P. caribaeorum.

In extranodalen Marginalzonen-B-Zell-Lymphomen vom MALT-Typ (eMZBCL) wurde fur verschiedene anatomische Lokalisationen eine Assoziation mit einer durch einen spezifischen Erreger verursachten chronischen Entzundung (z. B. Helicobacter pylori im Magen) beschrieben. Fur die Lunge ist eine solche Erregerassoziation bislang nicht bekannt. Mittels einer erstmalig auf diese Fragestellung angewandten sensitiven Screeningmethode (SHARP-Screening), die auf der Analyse der bakteriellen 16S-rRNA-Gene beruht, haben wir die mikrobielle Diversitat von 9 eMZBCL der Lunge analysiert und mit 9 Kontrollfallen verglichen. Interessanterweise konnten in 8 der 9 eMZBCL-Falle Bakterien der Alcaligenacea-Familie (Alcaligenes, Achromobacter, AKIW733 ) nachgewiesen werden, wahrend keiner der 9 Kontrollfalle eine Besiedlung mit diesen Keimen zeigte. Weitere Untersuchungen, z. B. mit spezifischen PRC-Assays an groseren Fallzahlen sind notig, um zu klaren, ob Betaproteobakterien der Alcaligenacea-Familie mit der Genese von eMZBCL in der Lunge assoziiert oder sogar ursachlich beteiligt sind.

The dichotomy between high microbial abundance (HMA) and low microbial abundance (LMA) sponges has been long recognized. In the present study, 56 sponge species from three geographic regions (greater Caribbean, Mediterranean, Red Sea) were investigated by transmission electron microscopy for the presence of microorganisms in the mesohyl matrix. Additionally, bacterial enumeration by DAPI-counting was performed on a subset of samples. Of the 56 species investigated, 28 were identified as belonging to the HMA and 28 to the LMA category. The sponge orders Agelasida and Verongida consisted exclusively of HMA species, and the Poecilosclerida were composed only of LMA sponges. Other taxa contained both types of microbial associations (e.g., marine Haplosclerida, Homoscleromorpha, Dictyoceratida), and a clear phylogenetic pattern could not be identified. For a few sponge species, an intermediate microbial load was determined, and the microscopy data did not suffice to reliably determine HMA or LMA status. To experimentally determine the HMA or LMA status of a sponge species, we therefore recommend a combination of transmission electron microscopy and 16S rRNA gene sequence data. This study significantly expands previous reports on microbial abundances in sponge tissues and contributes to a better understanding of the HMA-LMA dichotomy in sponge-microbe symbioses.

The aim of this study was to isolate bacteria with antimicrobial activities from the marine sponges Aplysina aerophoba and Aplysina cavernicola. The obtained 27 isolates could be subdivided into eight phylogenetically different clusters based on comparative sequence analysis of their 16S rDNA genes. The sponge isolates were affiliated with the low (Bacillus) and high G+C Gram-positive bacteria (Arthobacter, Micrococcus), as well as the alpha-Proteobacteria (unknown isolate) and gamma-Proteobacteria (Vibrio, Pseudoalteromonas). One novel Bacillus species was identified and two species were closely related to previously uncharacterized strains. Isolates with antimicrobial activity were numerically most abundant in the genera Pseudoalteromonas and the alpha-Proteobacteria. The sponge isolates show antimicrobial activities against Gram-positive and Gram-negative reference strains but not against the fungus Candida albicans. A general pattern was observed in that Gram-positive bacteria inhibited Gram-positive strains while Gram-negative bacteria inhibited Gram-negative isolates. Antimicrobial activities were also found against clinical isolates, i.e. multi-resistant Staphylococcus aureus and Staphylococcus epidermidis strains isolated from hospital patients. The high recovery of strains with antimicrobial activity suggests that marine sponges represent an ecological niche which harbors a hitherto largely uncharacterized microbial diversity and, concomitantly, a yet untapped metabolic potential.

Cultivated bacteria such as actinomycetes are a highly useful source of biomedically important natural products. However, such 'talented' producers represent only a minute fraction of the entire, mostly uncultivated, prokaryotic diversity. The uncultured majority is generally perceived as a large, untapped resource of new drug candidates, but so far it is unknown whether taxa containing talented bacteria indeed exist. Here we report the single-cell- and metagenomics-based discovery of such producers. Two phylotypes of the candidate genus 'Entotheonella' with genomes of greater than 9 megabases and multiple, distinct biosynthetic gene clusters co-inhabit the chemically and microbially rich marine sponge Theonella swinhoei. Almost all bioactive polyketides and peptides known from this animal were attributed to a single phylotype. 'Entotheonella' spp. are widely distributed in sponges and belong to an environmental taxon proposed here as candidate phylum 'Tectomicrobia'. The pronounced bioactivities and chemical uniqueness of 'Entotheonella' compounds provide significant opportunities for ecological studies and drug discovery.

Nature 506, 58–62 (2014); doi:10.1038/nature12959 One of the accession numbers for this Article was listed as AZHXW01000000 instead of AZHX01000000. It has been corrected in the online versions of the paper.

Marine sponges are a rich source of bioactive natural products and are promising sources for drug discovery and development An impressive example is the sponge Theonella swinhoei, which has yielded more than 120 compounds belonging to diverse structural types. Many sponges also harbor highly complex consortia of symbiotic bacteria that are suspected to be the true source of at least some of the secondary metabolites. In previous work, our group demonstrated a bacterial origin of onnamide- and psymberin-type polyketides for two different sponges (1–3), but there were no insights into the producer of compounds from other natural product families. In addition, the exact taxonomic identity of sponge-associated producers remained unknown. This talk will present new insights into these two issues. Isolation of genes encoding a peptide biosynthetic pathway from the T. swinhoei metagenome demonstrated a bacterial origin. Several genes were heterologously expressed and functionally characterized, which revealed unprecedented biosynthetic transformations. The novelty of these modifications suggests the existence of a structurally distinct natural product family, for which we propose the name proteusins. Using a strategy consisting of single-cell analysis and metagenomic sequencing, we identified the bacterial producer of onnamide polyketides in T. swinhoei. Surprisingly, the data suggest the symbiont to be a chemically exceptionally prolific bacterium, producing not only onnamides but most other compounds from this sponge chemotype, including the known and two previously unknown proteusins. Further biosynthetic studies and a survey of other sponges indicate that close relatives of the producer are widespread in these animals and vary with respect to their biosynthetic capabilities. These bacteria might therefore represent the first uncultivated taxon with a metabolic richness resembling that of major cultivated bacterial natural product sources. These results reveal a key role of symbiotic bacteria in the chemistry of their sponge hosts and provide new strategies to study uncultivated symbionts in a more systematic fashion.

Many marine sponges contain massive numbers of largely uncultivated, phylogenetically diverse bacteria that seem to be important contributors to the chemistry of these animals. Insights into the diversity, origin, distribution, and function of their metabolic gene communities are crucial to dissect the chemical ecology and biotechnological potential of sponge symbionts. This study reveals a sharp dichotomy between high and low microbial abundance sponges with respect to polyketide synthase (PKS) gene content, the presence of methyl-branched fatty acids, and the presence of members of the symbiotic candidate phylum "Poribacteria". For the symbiont-rich sponge Cacospongia mycofijiensis, a source of the tubulin-inhibiting fijianolides (=laulimalides), near-exhaustive large-scale sequencing of PKS gene-derived PCR amplicons was conducted. Although these amplicons exhibit high diversity at the sequence level, almost all of them belong to a single, architecturally unique group of PKSs present in "Poribacteria" and are proposed to synthesize methyl-branched fatty acids. Three components of this PKS were studied in vitro, providing initial insight into its biochemistry.

To provide insight into the phylogenetic bacterial diversity of the freshwater sponge Spongilla lacustris, a 16S rRNA gene libraries were constructed from sponge tissues and from lake water. Restriction fragment length polymorphism (RFLP) analysis of >190 freshwater sponge-derived clones resulted in six major restriction patterns, from which 45 clones were chosen for sequencing. The resulting sequences were affiliated with the Alphaproteobacteria (n = 19), the Actinobacteria (n = 15), the Betaproteobacteria (n = 2), and the Chloroflexi (n = 2) lineages. About half of the sequences belonged to previously described actinobacterial (hgc-I) and betaproteobacterial (beta-II) sequence clusters of freshwater bacteria that were also present in the lake water 16S rRNA gene library. At least two novel, deeply rooting alphaproteobacterial lineages were recovered from S. lacustris that showed

The recent application of molecular microbial ecology tools to sponge-microbe associations has revealed a glimpse into the biodiversity of these microbial communities, that is considered just ‘the tip of the iceberg’. This chapter provides an overview over these new findings with regard to identity, diversity and distribution patterns of sponge-associated microbial consortia. The sponges Aplysina aerophoba (Verongida), Rhopaloeides odorabile (Dicytoceratida) and Theonella swinhoei (Lithistida) were chosen as model systems for this review because they have been subject to both, cultivation-dependent and cultivation-independent approaches. A discussion of the microbial assemblages of Halichondria panicea is presented in the accompanying chapter by Imhoff and Stohr. Considering that a large fraction of sponge-associated microbes is not yet amenable to cultivation, an emphasis has been placed on the techniques centering around the 16S rRNA gene. A section has been included that covers the potential of sponge microbial communities for drug discovery. Finally, a ‘sponge-microbe interaction model’ is presented that summarizes our current understanding of the processes that might have shaped the community structure of the microbial assemblages within sponges.