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Applied and Environmental Microbiology, July 2006, p. 4775-4781, Vol. 72, No. 7
0099-2240/06/$08.00+0     doi:10.1128/AEM.00356-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Culture-Independent Characterization of the Digestive-Tract Microbiota of the Medicinal Leech Reveals a Tripartite Symbiosis

Paul L. Worthen, Cindy J. Gode,{dagger} and Joerg Graf*

Department of Molecular and Cell Biology, University of Connecticut, Storrs, Connecticut

Received 13 February 2006/ Accepted 6 May 2006

Culture-based studies of the microbial community within the gut of the medicinal leech have typically been focused on various Aeromonas species, which were believed to be the sole symbiont of the leech digestive tract. In this study, analysis of 16S rRNA gene clone libraries confirmed the presence of Aeromonas veronii and revealed a second symbiont, clone PW3, a novel member of the Rikenellaceae, within the crop, a large compartment where ingested blood is stored prior to digestion. The diversity of the bacterial community in the leech intestinum was determined, and additional symbionts were detected, including members of the {alpha}-, {gamma}-, and {delta}-Proteobacteria, Fusobacteria, Firmicutes, and Bacteroidetes. The relative abundances of the clones suggested that A. veronii and the novel clone, PW3, also dominate the intestinum community, while other clones, representing transient organisms, were typically present in low numbers. The identities of these transients varied greatly between individual leeches. Neither time after feeding nor feeding on defibrinated blood caused a change in identity of the dominant members of the microbial communities. Terminal restriction fragment length polymorphism analysis was used to verify that the results from the clone libraries were representative of a larger data set. The presence of a two-member bacterial community in the crop provides a unique opportunity to investigate both symbiont-symbiont and symbiont-host interactions in a natural model of digestive-tract associations.

* Corresponding author. Mailing address: Department of Molecular and Cell Biology, University of Connecticut, 91 N. Eagleville Rd., Unit 3125, Storrs, CT 06269-3125. Phone: (860) 486-9284. Fax: (860) 486-4331. E-mail: joerg.graf{at}uconn.edu.

{dagger} Present address: Department of Microbiology, University of Iowa, Iowa City, IA 52242.

Applied and Environmental Microbiology, July 2006, p. 4775-4781, Vol. 72, No. 7
0099-2240/06/$08.00+0     doi:10.1128/AEM.00356-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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