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Use of T-RFLPs for analyzing substrate attached bacteria in biofilms in the deep-waters of the Hellenic Trench (Ionian Sea). Christian-Albrechts-University of Kiel , FTZ, Hafentörn 1, Büsum, Germany Hellenic Centre for Marine Research , 46km Athens-Sounio, Anabyssos, Greece.
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Christian-Albrechts-University of Kiel, FTZ, Hafentörn 1, Büsum, Germany
Hellenic Centre for Marine Research , 46km Athens-Sounio, Anabyssos, Greece
Nikoleta Bellou, Evangelos Papathanassiou and Franciscus Colijn
Deepest part of the Mediterranean
Lies far from the effluents of major rivers
extremely clear water (light transmission length
55 + 10 m at 460 nm wavelength)
Weak water current consistently (below 10 cm/s)
Mean density of sea floor sediments 1.5 gr/cm3
slow accumulation rate of 7– 8cm over 10000 years
CTD profile at Nestor 4.5
First introduced by Zobell & Allen (1935) and Zobell & Anderson (1937)
Abundance and diversity regulated by physico-chemical factors
Bacteria prefer a sessile life form
Biofilms most commonly form as a result of some stress
Therefore, biofilms are found in many extreme environments
Assemblage of microbialcells
self-produced polymeric matrix
Within hours adhesion, colonization and growth of microbial populations
(Davis , 2007)
Biofilms on artificial substrata in deep-sea ?
Bacteria in biofilms ?
Comparison microbial communities in biofilms grown on different substrates & grown in different depths
Characterization of :
substrate relation within one deep-sea layer
depth relation for each substrate
depth and substrate relation
T-RFLP to compare microbial communities
PCR product + Restriction enzymes
PCR fluorescence (5’-FAM) labeled primer
Samples taken with a sterile swap
PCR amplifications of 16S rDNA showed that DNAisverifiable:
on each substrate
at all depth
Bacterial biofilm communities
do not differ highly between substrates within one depth
BUT differ along a depth gradient in one substrate type
Results indicate highly stratified biofilm communities
6thFramework Program of the European Commission to support the ‘European Research Area’