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Appl. Environ. Microbiol. doi:10.1128/AEM.01309-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Growth of environmental samples in a diffusion chamber increases the diversity of recovered isolates

Northeastern University, Department of Biology, 134 Mugar Life Science Building, 360 Huntington Avenue, Boston MA 2115, USA

^* To whom correspondence should be addressed. Email: s.epstein{at}neu.edu.

	Abstract

The majority of microorganisms from natural environments cannot be grown in the laboratory. The diffusion chamber-based approach is an alternative method that allows the microorganisms to grow their natural environment. An inoculum is sandwiched between semi-permeable (0.03-µm pore-size) membranes of the chamber, which is then returned into the source environment. The chamber allows for a free exchange of chemicals with the external milieu by diffusion, while restricting movement of cells. We used freshwater pond sediment to inoculate diffusion chambers and Petri dishes. The diffusion chambers were incubated on top of the sediment for four weeks. Both chamber- and Petri dish cultivation resulted in the isolation of numerous representatives of Alpha-, Beta-, and Gammaproteobacteria, Actinobacteria, Firmicutes and Bacteroidetes. However, the diffusion chamber-based approach also led to the isolation of species from rarely cultivated groups such as Deltaproteobacteria, Verrucomicrobia, Spirochaetes, and Acidobacteria. Material from the chambers was also transferred to new chambers in order to learn whether this will increase recovery of isolates. Several isolates could only be obtained from material transferred through multiple diffusion chambers. This suggests that continuous cultivation in diffusion chambers adapts some microorganisms for growth under otherwise prohibitive in vitro conditions.

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