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Applied and Environmental Microbiology, October 2007, p. 6386-6390, Vol. 73, No. 20
0099-2240/07/$08.00+0     doi:10.1128/AEM.01309-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Incubation of Environmental Samples in a Diffusion Chamber Increases the Diversity of Recovered Isolates{triangledown} ,{dagger}

Annette Bollmann,{ddagger} Kim Lewis, and Slava S. Epstein*

Northeastern University, Department of Biology, 134 Mugar Life Science Building, 360 Huntington Avenue, Boston, Massachusetts 02115

Received 12 June 2007/ Accepted 15 August 2007

The majority of microorganisms from natural environments cannot be grown in the laboratory. The diffusion-chamber-based approach is an alternative method that allows microorganisms to grow in their natural environment. An inoculum is sandwiched between semipermeable (0.03-µm-pore-size) membranes of the chamber, which is then returned to the source environment. The chamber allows for a free exchange of chemicals with the external milieu by diffusion while restricting the 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 4 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 the recovery of isolates. Several isolates could be obtained only 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.

* Corresponding author. Mailing address: Northeastern University, Department of Biology, 134 Mugar Life Science Building, 360 Huntington Ave., Boston, MA 02115. Phone: (617) 373-4048. Fax: (617) 373-3724. E-mail: s.epstein{at}neu.edu

{triangledown} Published ahead of print on 24 August 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

{ddagger} Present address: Miami University, Department of Microbiology, 32 Pearson Hall, Oxford, OH 45056.

Applied and Environmental Microbiology, October 2007, p. 6386-6390, Vol. 73, No. 20
0099-2240/07/$08.00+0     doi:10.1128/AEM.01309-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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