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. 2024 Sep 26;22(10):440.
doi: 10.3390/md22100440.

Diversity and Activity of Bacteria Cultured from a Cup-The Sponge Calyx nicaeensis

Lynne Itelson  1 , Mayan Merav  2 , Shai Haymi  1 , Shmuel Carmeli  2 , Micha Ilan  1
Affiliations

Diversity and Activity of Bacteria Cultured from a Cup-The Sponge Calyx nicaeensis

Lynne Itelson et al. Mar Drugs. .

Abstract

Marine sponges are well-known for hosting rich microbial communities. Sponges are the most prolific source of marine bioactive compounds, which are frequently synthesized by their associated microbiota. Calyx nicaeensis is an endemic Mediterranean sponge with scarce information regarding its (bioactive) secondary metabolites. East Mediterranean specimens of mesophotic C. nicaeensis have never been studied. Moreover, no research has inspected its associated bacteria. Thus, we studied the sponge's bacterial diversity and examined bacterial interspecific interactions in search of a promising antibacterial candidate. Such novel antimicrobial agents are needed since extensive antibiotic use leads to bacterial drug resistance. Bacteria cultivation yielded 90 operational taxonomic units (OTUs). A competition assay enabled the testing of interspecific interactions between the cultured OTUs. The highest-ranked antagonistic bacterium, identified as Paenisporosarcina indica (previously never found in marine or cold habitats), was mass cultured, extracted, and separated using size exclusion and reversed-phase chromatographic methods, guided by antibacterial activity. A pure compound was isolated and identified as 3-oxy-anteiso-C15-fatty acid-lichenysin. Five additional active compounds await final cleaning; however, they are lichenysins and surfactins. These are the first antibacterial compounds identified from either the C. nicaeensis sponge or P. indica bacterium. It also revealed that the genus Bacillus is not an exclusive producer of lichenysin and surfactin.

Keywords: Calyx nicaeensis; Paenisporosarcina indica; antibiotics; biosurfactants; mesophotic; sponges.

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Conflict of interest statement

The authors declare no conflicts of interest. The funders had no role in the design of this study; the collection, analyses, or interpretation of data; the writing of the manuscript; or the decision to publish the results.

Figures

Figure 1
Figure 1
Calyx nicaeensis sponge (in situ at 99 m). Photo by Micha Ilan’s Lab.
Figure 2
Figure 2
Distribution of the sponge-associated OTUs cultivated from C. nicaeensis per genus. Genera above 3% are named in this figure.
Figure 3
Figure 3
Growth inhibition and promotion effects of 18 C. nicaeensis-associated OTUs tested in the competition assay. (_)—inhibition, (_)—promotion.
Figure 4
Figure 4
Interspecific interactions assessed using eigenvector centrality between the cultivated OTUs from the C. nicaeensis sponge. Numbers represent different OTUs. Larger nodes represent the eigenvector centrality score. Different colors mark bacteria that inhibit a large number of bacteria. Green arrows represent inhibition >1 cm and purple >1 cm. The four OTUs with the strongest inhibitions were 83, 72, 43, and 10.
Figure 5
Figure 5
Lichenysins and surfactins from Paenisporosarcina indica associated with the marine sponge Calyx nicaeensis.
Figure 6
Figure 6
A schematic representation of a competition assay. A—The tested bacterium along the center line. B—Different OTUs smeared vertically with 1.2 cm between them, without touching the vertical line.
Figure 7
Figure 7
Disk diffusion assay. MeOH, DCM, and bacteria cell mass extracts on filters. NC—negative control. Extracts show antibacterial activity (halo), while the NC does not.
See this image and copyright information in PMC

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