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Applied and Environmental Microbiology, May 2006, p. 3161-3167, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3161-3167.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Wolbachia Infections in the Cimicidae: Museum Specimens as an Untapped Resource for Endosymbiont Surveys

Joyce M. Sakamoto,1 Julie Feinstein,2 and Jason L. Rasgon3*

Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, Maryland 21201,1 Ambrose Monell Collection for Molecular and Microbial Research, Division of Invertebrate Zoology, American Museum of Natural History, New York, New York 10024,2 The W. Harry Feinstone Department of Molecular Microbiology and Immunology, The Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, Maryland 212053

Received 7 September 2005/ Accepted 24 February 2006

Wolbachia spp. are obligate maternally inherited endosymbiotic bacteria that infect diverse arthropods and filarial nematodes. Previous microscopic and molecular studies have identified Wolbachia in several bed bug species (Cimicidae), but little is known about how widespread Wolbachia infections are among the Cimicidae. Because cimicids of non-medical importance are not commonly collected, we hypothesized that preserved museum specimens could be assayed for Wolbachia infections. For the screening of museum specimens, we designed a set of primers that specifically amplify small diagnostic fragments (130 to 240 bp) of the Wolbachia 16S rRNA gene. Using these and other previously published primers, we screened 39 cimicid species (spanning 16 genera and all 6 recognized subfamilies) and 2 species of the sister family Polyctenidae for Wolbachia infections using museum and wild-caught material. Amplified fragments were sequenced to confirm that our primers were amplifying Wolbachia DNA. We identified 10 infections, 8 of which were previously undescribed. Infections in the F supergroup were common in the subfamily Cimicinae, while infections in the A supergroup were identified in the subfamilies Afrocimicinae and Haematosiphoninae. Even though specimens were degraded, we detected infections in over 23% of cimicid species. Our results indicate that Wolbachia infections may be common among cimicids and that archived museum material is a useful untapped resource for invertebrate endosymbiont surveys. The new screening primers listed in this report will be useful for other researchers conducting Wolbachia surveys with specimens with less-than-optimum DNA quality.

* Corresponding author. Mailing address: The W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, E4626, 615 N. Wolfe Street, Baltimore, MD 21205. Phone: (410) 502-2584. Fax: (410) 955-0105. E-mail: jrasgon{at}jhsph.edu.

Applied and Environmental Microbiology, May 2006, p. 3161-3167, Vol. 72, No. 5
0099-2240/06/$08.00+0     doi:10.1128/AEM.72.5.3161-3167.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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