Microbial Ecology

, Volume 65, Issue 2, pp 424–436 | Cite as

Adaptation of the 3H-Leucine Incorporation Technique to Measure Heterotrophic Activity Associated with Biofilm on the Blades of the Seaweed Sargassum spp.

  • Sergio A. Coelho-Souza
  • Marcio R. Miranda
  • Leonardo T. Salgado
  • Ricardo Coutinho
  • Jean R. D. Guimaraes


The ecological interaction between microorganisms and seaweeds depends on the production of secondary compounds that can influence microbial diversity in the water column and the composition of reef environments. We adapted the 3H-leucine incorporation technique to measure bacterial activity in biofilms associated with the blades of the macroalgae Sargassum spp. We evaluated (1) if the epiphytic bacteria on the blades were more active in detritus or in the biofilm, (2) substrate saturation and linearity of 3H-leucine incorporation, (3) the influence of specific metabolic inhibitors during 3H-leucine incorporation under the presence or absence of natural and artificial light, and (4) the efficiency of radiolabeled protein extraction. Scanning electron microscopy showed heterogeneous distribution of bacteria, diatoms, and polymeric extracellular secretions. Active bacteria were present in both biofilm and detritus on the blades. The highest 3H-leucine incorporation was obtained when incubating blades not colonized by macroepibionts. Incubations done under field conditions reported higher 3H-leucine incorporation than in the laboratory. Light quality and sampling manipulation seemed to be the main factors behind this difference. The use of specific metabolic inhibitors confirmed that bacteria are the main group incorporating 3H-leucine but their association with primary production suggested a symbiotic relationship between bacteria, diatoms, and the seaweed.


Detritus Extracellular Polymeric Substance Diuron Incorporation Rate Leucine Incorporation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We thank the IEAPM logistic support, especially to the Grupo de Quimica, the Department of Oceanography, Division of Biotechnology Lab team, and LEG 10 team. This study would not have been concluded without the help of Maria Helena Baeta-Neves, Antônio Casarin, Dagles Viana dos Reis, William Romão, and Carlos Eduardo Leite Ferreira. We also thank Dr. Stuart Jenkins and the anonymous referees who contributed to the final format of this manuscript. This work was supported by CAPES, CNPq, and FAPERJ.


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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Sergio A. Coelho-Souza
    • 1
    • 2
    • 3
    • 6
  • Marcio R. Miranda
    • 3
    • 4
  • Leonardo T. Salgado
    • 5
  • Ricardo Coutinho
    • 1
  • Jean R. D. Guimaraes
    • 4
  1. 1.Biotecnologia MarinhaInstituto de Ciências do Mar Almirante Paulo Moreira (IEAPM)Arraial do Cabo-RJBrazil
  2. 2.Universidade Federal do Rio de Janeiro (PPGE/UFRJ)Rio de JaneiroBrazil
  3. 3.INPeTAm/IBCCFUniversidade Federal do Rio de Janeiro (UFRJ)Rio de Janeiro-RJBrazil
  4. 4.LTWCP/IBCCFUniversidade Federal do Rio de Janeiro (UFRJ)Rio de Janeiro-RJBrazil
  5. 5.DIPEQInstituto de Pesquisas Jardim Botânico do Rio de Janeiro (IPJBRJ)Rio de Janeiro-RJBrazil
  6. 6.Instituto de Biofísica Carlos Chagas FilhoCentro de Ciências e SaúdeRio de JaneiroBrazil

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