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Appl. Environ. Microbiol. doi:10.1128/AEM.00537-07
Copyright (c) 2007, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

Localization and Visualization of a Coxiella-type Symbiont within the Lone Star Tick Amblyomma americanum

Department of Biology, Indiana University, Bloomington, IN 47405

^* To whom correspondence should be addressed. Email: cfuqua{at}indiana.edu.

	Abstract

A Coxiella-type microbe occurs at 100% frequency in all Amblyomma americanum ticks thus far tested (Amblyomma-associated Coxiella – AAC). Using laboratory-reared ticks free of other microbes, we identified the AAC microbe in several A. americanum tissues and life history stages by 16S rRNA gene sequencing and diagnostic PCR. We visualized AAC through the use of a diagnostic fluorescence in situ hybridization (FISH) assay supplemented with PCR-based detection, nucleic acid fluorescent staining, wide-field epifluorescence and confocal microscopy, and transmission electron microscopy (TEM). Specific fluorescent foci were observed in several tick tissues, including the midgut and the Malpighian tubules, but particularly bright signals were observed in the granular acini of salivary gland clusters and in both small and large oocytes. TEM confirmed intracellular bacterial structures in the same tissues. The presence of AAC within oocytes is consistent with the vertical transmission of these endosymbionts. Further, the presence of the AAC symbiont in other tissues such as salivary glands could potentially lead to interactions with horizontally-acquired pathogens.