, Volume 22, Issue 4, pp 773–795 | Cite as

Molecular assessment of microbiota structure and dynamics along mixed olive oil and winery wastewaters biotreatment

  • Ana Eusébio
  • Marta Tacão
  • Sandra Chaves
  • Rogério Tenreiro
  • Elsa Almeida-Vara
Original Paper


The major parcel of the degradation occurring along wastewater biotreatments is performed either by the native microbiota or by added microbial inocula. The main aim of this study was to apply two fingerprinting methods, temperature gradient gel electrophoresis (TGGE) and length heterogeneity-PCR (LH-PCR) analysis of 16S rRNA gene fragments, in order to assess the microbiota structure and dynamics during mixed olive oil and winery wastewaters aerobic biotreatment performed in a jet-loop reactor (JLR). Sequence homology analysis showed the presence of bacterial genera Gluconacetobacter, Klebsiella, Lactobacillus, Novosphingobium, Pseudomonas, Prevotella, Ralstonia, Sphingobium and Sphingomonas affiliated with five main phylogenetic groups: alpha-, beta- and gamma-Proteobacteria, Firmicutes and Bacteroidetes. LH-PCR analysis distinguished eight predominant DNA fragments correlated with the samples showing highest performance (COD removal rates of 67 up to 75%). Cluster analysis of both TGGE and LH-PCR fingerprinting profiles established five main clusters, with similarity coefficients higher than 79% (TGGE) and 62% (LH-PCR), and related with hydraulic retention time, indicating that this was the main factor responsible for the shifts in the microbiota structure. Canonical correspondence analysis revealed that changes observed on temperature and O2 level were also responsible for shifts in microbiota composition. Community level metabolic profile analysis was used to test metabolic activities in samples. Integrated data revealed that the microbiota structure corresponds to bacterial groups with high degradative potential and good suitability for this type of effluents biotreatments.


Microbiota structure Microbiota dynamics Microbiota diversity TGGE LH-PCR Olive oil wastewater Winery wastewater 



This work was supported by the FCT Project MOTIVE (PPCDT/AMB/56616/2004). Sandra Chaves was a FCT post-doc fellow (BPD/20819/2004). Authors wish to thank José Cardoso Duarte from LNEG (Unidade de Bioenergia) for jet-loop reactor availability.


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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Ana Eusébio
    • 1
    • 2
  • Marta Tacão
    • 1
  • Sandra Chaves
    • 3
  • Rogério Tenreiro
    • 3
  • Elsa Almeida-Vara
    • 2
  1. 1.LNEG, Unidade de Bioenergia, Estrada do Paço do LumiarLisbonPortugal
  2. 2.Centro de Biodiversidade, Genómica Integrativa e Funcional (BioFIG), FCLLisbonPortugal
  3. 3.Universidade de Lisboa, Faculdade de Ciências, Centro de Biodiversidade, Genómica Integrativa e Funcional (BioFIG), Edifício ICATLisbonPortugal

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