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A method for the use of the radiotracer 55Fe for microautoradiography and CARD-FISH of natural bacterial communities.
MedLine Citation:
PMID:  23050807     Owner:  NLM     Status:  Publisher    
Abstract/OtherAbstract:
Heterotrophic bacteria are key players in the biogeochemical cycle of iron (Fe) in the ocean, but the capability of different bacterial groups to access this micronutrient is ignored thus far. The aim of our study was to develop a protocol for the combined application of microautoradiography (MICRO) and Catalyzed Reporter Deposition-Fluorescence In Situ Hybridization (CARD-FISH) using the radioisotope (55) Fe. Among the different washing solutions tested, Ti-citrate-EDTA was the most efficient for the removal of extracellular (55) Fe providing sufficiently low background values. We further demonstrate that the washing of cells with Ti-citrate-EDTA, and the fixation with paraformaldehyde or formaldehyde does not induce leakage of intracellular (55) Fe. Incubating natural bacterial communities collected from contrasting environments, the NW Mediterranean Sea and the Southern Ocean, with (55) Fe revealed that 3% to 29% of bacterial cells were associated with silver grains. Combining microautoradiography with CARD-FISH, we demonstrate that the contribution of different bacterial groups to total (55) Fe incorporating cells was overall reflected by their relative contribution to abundance. An exception to this pattern was the proportionally higher contribution of Gammaproteobacteria, SAR86 and Alteromonas. Our study demonstrates the feasibility of MICRO-CARD-FISH using the radiotracer (55) Fe and provides the first description of marine bacterial assemblages actively incorporating Fe. © 2012 Federation of European Microbiological Societies. Published by Blackwell PublishingLtd. All rights reserved.
Authors:
Marion Fourquez; Ingrid Obernosterer; Stéphane Blain
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Publication Detail:
Type:  LETTER     Date:  2012-10-11
Journal Detail:
Title:  FEMS microbiology letters     Volume:  -     ISSN:  1574-6968     ISO Abbreviation:  FEMS Microbiol. Lett.     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-10-11     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7705721     Medline TA:  FEMS Microbiol Lett     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Copyright Information:
© 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.
Affiliation:
Univ paris 06, CNRS, Laboratoire d'Océanographie Microbienne, Observatoire océanologique, avenue du Fontaulé, 66650, Banyuls-sur-mer, France.
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