The filtration–acclimatization method for isolation of an important fraction of the not readily cultivable bacteria

We developed a novel method, the filtration–acclimatization method (FAM), which enables the isolation and cultivation of an important fraction of the bacterial diversity, which is not cultivable by standard methods. The method consists of a filtration step, which removes most of the readily cultivab...

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Bibliographic Details
Published in:Journal of microbiological methods 2004-06, Vol.57 (3), p.379-390
Main Authors: Hahn, Martin W, Stadler, Peter, Wu, Qinglong L, Pöckl, Matthias
Format: Article
Language:English
Subjects:
0.2 μm filtration
Actinobacteria
Bacteria - genetics
Bacteria - growth & development
Bacteria - isolation & purification
Bacteriological methods and techniques used in bacteriology
Bacteriology
Bacteroidetes
Base Sequence
Biological and medical sciences
Culture Media
DNA, Bacterial - chemistry
DNA, Bacterial - genetics
Filtration - methods
Filtration–acclimatization method
Fresh Water - microbiology
Freshwater bacterioplankton
Fundamental and applied biological sciences. Psychology
Isolation of bacteria
Microbiology
Microscopy, Fluorescence
Molecular Sequence Data
Phylogeny
Polymerase Chain Reaction
Proteobacteria
RNA, Ribosomal, 16S - chemistry
RNA, Ribosomal, 16S - genetics
Sequence Alignment
Sequence Analysis, DNA
Spirochaeta
Ultramicrobacteria
Water Microbiology
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Summary:We developed a novel method, the filtration–acclimatization method (FAM), which enables the isolation and cultivation of an important fraction of the bacterial diversity, which is not cultivable by standard methods. The method consists of a filtration step, which removes most of the readily cultivable bacteria able to overgrow slowly growing bacteria, and an acclimatization procedure that provides a slow transition from the low environmental substrate concentrations to the high concentration of standard microbial media. So far, we isolated in total 65 strains from surface freshwater habitats by utilizing FAM. The isolates are affiliated with Actinobacteria, Alpha-, Betaproteobacteria, Bacteroidetes, and Spirochaeta. All isolates are pure cultures and form visible colonies on agar plates with high substrate concentrations. For further analysis, strains sharing more than a 97% 16S rRNA gene sequence similarity were grouped into one taxon. Based on sequence similarities, 88% of the obtained taxa can be considered to be undescribed species (
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2004.02.004