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  4. Application of the denaturing gradient gel electrophoresis (DGGE) technique as an efficient diagnostic tool for ciliate communities in soil
 
research article

Application of the denaturing gradient gel electrophoresis (DGGE) technique as an efficient diagnostic tool for ciliate communities in soil

Jousset, Alexandre
•
Lara, Enrique
•
Nikolausz, Marcell
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2010
Science Of The Total Environment

Ciliates (or Ciliophora) are ubiquitous organisms which can be widely used as bioindicators in ecosystems exposed to anthropogenic and industrial influences. The evaluation of the environmental impact on soil ciliate communities with methods relying on morphology-based identification may be hampered by the large number of samples usually required for a statistically supported, reliable conclusion. Cultivation-independent molecular-biological diagnostic tools are a promising alternative to greatly simplify and accelerate such studies. In this present work a ciliate-specific fingerprint method based on the amplification of a phylogenetic marker gene (i.e. the 18S ribosomal RNA gene) with subsequent analysis by denaturing gradient gel electrophoresis (DGGE) was developed and used to monitor community shifts in a polycyclic aromatic hydrocarbon (PAH) polluted soil. The semi-nested approach generated ciliate-specific amplification products from all soil samples and allowed to distinguish community profiles from a PAH-polluted and a non-polluted control soil. Subsequent sequence analysis of excised bands provided evidence that polluted soil samples are dominated by organisms belonging to the class Colpodea. The general DGGE approach presented in this study might thus in principle serve as a fast and reproducible diagnostic tool, complementing and facilitating future ecological and ecotoxicological monitoring of ciliates in polluted habitats. (c) 2009 Elsevier B.V. All rights reserved.

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Type
research article
DOI
10.1016/j.scitotenv.2009.09.056
Web of Science ID

WOS:000274365000027

Author(s)
Jousset, Alexandre
Lara, Enrique
Nikolausz, Marcell
Harms, Hauke
Chatzinotas, Antonis
Date Issued

2010

Published in
Science Of The Total Environment
Volume

408

Start page

1221

End page

1225

Subjects

Ciliates

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Dgge

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18S rRNA

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Soil

•

Monitoring

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Pollution

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Pah

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Subunit Ribosomal-Rna

•

Annealing Temperature

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Eukaryotic Diversity

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Primer Mismatch

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Pcr Primers

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Protozoa

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Ciliophora

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Sequences

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Phylogeny

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Emphasis

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
IIE  
Available on Infoscience
December 16, 2011
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/75731
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