This Article Full Text Full Text (PDF) Other Versions of this Article: AEM.01673-06v1 72/11/6934    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Citing Articles Citing Articles via Google Scholar Google Scholar Articles by Rasgon, J. L. Articles by Ren, X. Search for Related Content PubMed PubMed Citation Articles by Rasgon, J. L. Articles by Ren, X. Agricola Articles by Rasgon, J. L. Articles by Ren, X.

Survival of Wolbachia pipientis in Cell-Free Medium

Jason L. Rasgon,^1,2* Courtney E. Gamston,^1, and Xiaoxia Ren^1,2,

The W. Harry Feinstone Department of Molecular Microbiology and Immunology, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland 21205,¹ The Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland 21205²

Received 18 July 2006/ Accepted 28 August 2006

Wolbachia pipientis is an obligate intracellular bacterium found in a wide range of invertebrate taxa. While over ecological timescales Wolbachia infections are maintained by strict maternal inheritance, horizontal transfer events are common over evolutionary time. To be horizontally transferred between organisms, Wolbachia bacteria must pass through and survive an extracellular phase. We used BacLight live-dead staining, PCR, and fluorescence in situ hybridization to assess the ability for purified Wolbachia bacteria to survive in cell-free media. We found that purified Wolbachia bacteria were able to survive extracellularly for up to 1 week with no decrease in viability. While no replication was observed in the extracellular phase, purified Wolbachia bacteria were able to reinvade cells and establish stable infections at all time points. The ability of Wolbachia bacteria to survive outside host cells may increase the probability of successful horizontal transfer and the exploitation of new ecological niches. Our development of methods to purify and maintain viable Wolbachia bacteria from cultured cells will be useful for other researchers studying Wolbachia biology.

Published ahead of print on 1 September 2006.

C.E.G. and X.R. contributed equally to this work.