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. 2015 May 7:6:389.
doi: 10.3389/fmicb.2015.00389. eCollection 2015.

The antimicrobial activity of heterotrophic bacteria isolated from the marine sponge Erylus deficiens (Astrophorida, Geodiidae)

Ana Patrícia Graça  1 , Flávia Viana  2 , Joana Bondoso  1 , Maria Inês Correia  3 , Luis Gomes  3 , Madalena Humanes  4 , Alberto Reis  5 , Joana R Xavier  6 , Helena Gaspar  4 , Olga M Lage  1
Affiliations

The antimicrobial activity of heterotrophic bacteria isolated from the marine sponge Erylus deficiens (Astrophorida, Geodiidae)

Ana Patrícia Graça et al. Front Microbiol. .

Abstract

Interest in the study of marine sponges and their associated microbiome has increased both for ecological reasons and for their great biotechnological potential. In this work, heterotrophic bacteria associated with three specimens of the marine sponge Erylus deficiens, were isolated in pure culture, phylogenetically identified and screened for antimicrobial activity. The isolation of bacteria after an enrichment treatment in heterotrophic medium revealed diversity in bacterial composition with only Pseudoalteromonas being shared by two specimens. Of the 83 selected isolates, 58% belong to Proteobacteria, 23% to Actinobacteria and 19% to Firmicutes. Diffusion agar assays for bioactivity screening against four bacterial strains and one yeast, revealed that a high number of the isolated bacteria (68.7%) were active, particularly against Candida albicans and Vibrio anguillarum. Pseudoalteromonas, Microbacterium, and Proteus were the most bioactive genera. After this preliminary screening, the bioactive strains were further evaluated in liquid assays against C. albicans, Bacillus subtilis and Escherichia coli. Filtered culture medium and acetone extracts from three and 5 days-old cultures were assayed. High antifungal activity against C. albicans in both aqueous and acetone extracts as well as absence of activity against B. subtilis were confirmed. Higher levels of activity were obtained with the aqueous extracts when compared to the acetone extracts and differences were also observed between the 3 and 5 day-old extracts. Furthermore, a low number of active strains was observed against E. coli. Potential presence of type-I polyketide synthases (PKS-I) and non-ribosomal peptide synthetases (NRPSs) genes were detected in 17 and 30 isolates, respectively. The high levels of bioactivity and the likely presence of associated genes suggest that Erylus deficiens bacteria are potential sources of novel marine bioactive compounds.

Keywords: Candida albicans; Erylus deficiens; PKS-I and NRPS genes; bioactivity screening assay; sponge associated heterotrophic bacteria.

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Figures

Figure 1
Figure 1
Phylogenetic 16S rRNA gene tree generated by maximum-likelihood analysis based in General Time Reversible model and Gamma distributed with Invariant sites (G+I) indicating the relationship of the heterotrophic bacteria isolated from three E. deficiens specimens. Thermatoga maritima was used as outgroup. Bar – 0.05 substitutions per 100 nucleotides. *Presence of NRPS genes; **Presence of PKS-I genes; ***Presence of both genes.
Figure 2
Figure 2
Relation between the hits of bioactive genera and the target organisms studied in the preliminary screening.
Figure 3
Figure 3
Comparison of the levels of inhibition (higher than 25%) between the aqueous and the acetone extracts from Erylus deficiens isolates in the liquid assays. The numbers above each box represent the number of bioactive extracts.
Figure 4
Figure 4
Levels of inhibition (higher than 25%) of 3 and 5 days and aqueous (aqu) and acetone (acet) extracts from Erylus deficiens isolates against C. albicans (Ca) and E. coli (Ec). No result is shown for aqueous, 3 days extracts against E. coli due to the absence of inhibition. The numbers above each box represent the number of bioactive extracts.
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