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Applied and Environmental Microbiology, May 2009, p. 3331-3343, Vol. 75, No. 10
0099-2240/09/$08.00+0     doi:10.1128/AEM.02101-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Diversity of Bacteria in the Marine Sponge Aplysina fulva in Brazilian Coastal Waters{triangledown} ,{dagger}

C. C. P. Hardoim,1,{ddagger} R. Costa,1,2,{ddagger} F. V. Araújo,3 E. Hajdu,4 R. Peixoto,5 U. Lins,6 A. S. Rosado,5 and J. D. van Elsas1*

Department of Microbial Ecology, Centre for Ecological and Evolutionary Studies, University of Groningen, Kerklaan 30, Groningen 9751 NN, The Netherlands,1 Centre for Marine Sciences (CCMAR-CIMAR), University of Algarve, Gambelas, 8005-139 Faro, Portugal,2 Faculdade de Formação de Professores, Universidade Estadual do Rio de Janeiro, Francisco Portela 1470, Patronato, São Gonçalo, Rio de Janeiro, CEP 24435-000, Brazil,3 Laboratório de Porífera, Departamento de Invertebrados, Museu Nacional, Universidade Federal do Rio de Janeiro, Quinta da Boa Vista, s/n, São Cristóvão, Rio de Janeiro, CEP 20940-040, Brazil,4 Laboratório de Ecologia Microbiana Molecular,5 Laboratório de Ultraestrutura de Procariotos, Instituto de Microbiologia Prof. Paulo de Góes, Universidade Federal do Rio de Janeiro, Centro de Ciências da Saúde, Bloco I Cidade Universitária, Ilha do Fundão, Rio de Janeiro, CEP 21.941-590, Brazil6

Received 11 September 2008/ Accepted 12 March 2009

Microorganisms can account for up to 60% of the fresh weight of marine sponges. Marine sponges have been hypothesized to serve as accumulation spots of particular microbial communities, but it is unknown to what extent these communities are directed by the organism or the site or occur randomly. To address this question, we assessed the composition of specific bacterial communities associated with Aplysina fulva, one of the prevalent sponge species inhabiting Brazilian waters. Specimens of A. fulva and surrounding seawater were collected in triplicate in shallow water at two sites, Caboclo Island and Tartaruga beach, Búzios, Brazil. Total community DNA was extracted from the samples using "direct" and "indirect" approaches. 16S rRNA-based PCR-denaturing gradient gel electrophoresis (PCR-DGGE) analyses of the total bacterial community and of specific bacterial groups—Pseudomonas and Actinobacteria—revealed that the structure of these assemblages in A. fulva differed drastically from that observed in seawater. The DNA extraction methodology and sampling site were determinative for the composition of actinobacterial communities in A. fulva. However, no such effects could be gleaned from total bacterial and Pseudomonas PCR-DGGE profiles. Bacterial 16S rRNA gene clone libraries constructed from directly and indirectly extracted DNA did not differ significantly with respect to diversity and composition. Altogether, the libraries encompassed 15 bacterial phyla and the candidate division TM7. Clone sequences affiliated with the Cyanobacteria, Chloroflexi, Gamma- and Alphaproteobacteria, Actinobacteria, Bacteroidetes, and Acidobacteria were, in this order, most abundant. The bacterial communities associated with the A. fulva specimens were distinct and differed from those described in studies of sponge-associated microbiota performed with other sponge species.

* Corresponding author. Mailing address: Department of Microbial Ecology, Centre for Ecological and Evolutionary Studies, University of Groningen, Kerklaan 30, Groningen 9751 NN, The Netherlands. Phone: (31) 50 363 2151. Fax: (31) 50 363 2154. E-mail: j.d.van.elsas{at}rug.nl

{triangledown} Published ahead of print on 20 March 2009.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

{ddagger} C.C.P.H. and R.C. contributed equally to this study.

Applied and Environmental Microbiology, May 2009, p. 3331-3343, Vol. 75, No. 10
0099-2240/09/$08.00+0     doi:10.1128/AEM.02101-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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