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Application of denaturing gradient gel electrophoresis for analysing the gut microflora of Lumbricus rubellus Hoffmeister under different feeding conditions

Published online by Cambridge University Press:  28 April 2008

B.A. Knapp ,
J. Seeber ,
S.M. Podmirseg ,
E. Meyer  and
H. Insam
B.A. Knapp*
Affiliation:
University of Innsbruck, Institute of Microbiology, Technikerstrasse 25, 6020 Innsbruck, Austria
J. Seeber
Affiliation:
University of Innsbruck, Institute of Ecology, Technikerstrasse 25, 6020 Innsbruck, Austria
S.M. Podmirseg
Affiliation:
University of Innsbruck, Institute of Microbiology, Technikerstrasse 25, 6020 Innsbruck, Austria
E. Meyer
Affiliation:
University of Innsbruck, Institute of Ecology, Technikerstrasse 25, 6020 Innsbruck, Austria
H. Insam
Affiliation:
University of Innsbruck, Institute of Microbiology, Technikerstrasse 25, 6020 Innsbruck, Austria
*
*Author for correspondence Fax: 00435125072928 E-mail: [email protected]
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Abstract

The earthworm, Lumbricus rubellus, plays an essential role in soil ecosystems as it affects organic matter decomposition and nutrient cycling. By ingesting a mixture of organic and mineral material, a variety of bacteria and fungi are carried to the intestinal tract of the earthworm. To get a better understanding of the interactions between L. rubellus and the microorganisms ingested, this study tried to reveal if the diet affects the composition of the gut microflora of L. rubellus or if its intestinal tract hosts an indigenous, species-specific microbiota. A feeding experiment with L. rubellus was set up; individuals were collected in the field, transferred to a climate chamber and fed with food sources of different quality (dwarf shrub litter, grass litter or horse dung) for six weeks. DNA was extracted from the guts of the earthworms, as well as from the food sources and the surrounding soil, and further analysed by a molecular fingerprinting method, PCR-DGGE (Polymerase Chain Reaction – Denaturing Gradient Gel Electrophoresis). We were able to demonstrate that the gut microbiota was strongly influenced by the food source ingested and was considerably different to that of the surrounding soil. Sequencing of dominant bands of the bacterial DGGE fingerprints revealed a strong occurrence of y-Proteobacteria in all gut samples, independent of the food source. A specific microflora in the intestinal tract of L. rubellus, robust against diet changes, could not be found.

Keywords

Lumbricus rubellus Hoffmeisterpolymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysisgut microbiotamicrobial community compositionband identification
Type
Research Paper
Information
Bulletin of Entomological Research , Volume 98 , Issue 3 , June 2008 , pp. 271 - 279
Copyright
Copyright © 2008 Cambridge University Press

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