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FINDING NEEDLES IN THE HAYSTACK: SINGLE COPY MICROSATELLITE LOCI FOR AEDES JAPONICUS (DIPTERA: CULICIDAE)

First identified in three North American states in 1998, Aedes japonicus japonicus, the Asian bush mosquito, has since spread to 21 states, plus Ontario in Canada, northern France, and Belgium. Analyses of the introduction and expansion of this potentially deadly disease vector will be radically improved by including powerful genetic markers like microsatellites. Useful microsatellite loci have, however, been difficult to identify for mosquitoes in the genus Aedes because of the high amount of repetitive DNA in these species. We isolated single-copy DNA from Ae. j. japonicus and then used a standard enrichment method to identify regions containing microsatellites. Here we describe seven polymorphic microsatellite loci that were tested in American populations of Ae. j. japonicus. These loci were also found to be polymorphic in two other of the four Ae. japonicus subspecies and in Aedes koreicus.

Received January 27, 2005. Accepted for publication May 19, 2005.

Acknowledgments: We are indebted to Nusha Keyghobadi for proposing the low-copy DNA protocol to develop microsatellites for Aedes japonicus; Sven Erik Spilchiger and Michael Hutchinson (PA-DEP) for providing us with specimens of Aedes japonicus japonicus from eastern Pennsylvania; Drs. Ichiro Miyagi and Takako Toma for specimens of related subspecies and species in Japan; Heung-Chul Kim for Aedes koreicus from Korea; Peter Armbruster for specimens of Aedes albopictus and Ae. aegypti from Florida; Julie Smith, Kenli Okada, Carolyn Bahnck, Erika Butler, and an anonymous reviewer for comments on an earlier version of the manuscript; Tapan Ganguly and the DNA Sequencing Facility, University of Pennsylvania, for technical assistance.

Financial support: This project has been funded in whole with federal funds from the National Institute of Allergy and Infectious Disease, National Institutes of Health, under contract no. N01-AI-25490.

^* Address correspondence to Dina M. Fonseca, Academy of Natural Sciences, 1900 Ben Franklin Parkway, Philadelphia, PA 19103. E-mail: fonseca{at}acnatsci.org

Author’s addresses: Andrea K. Widdel, Academy of Natural Sciences, 1900 Ben Franklin Parkway, Philadelphia, PA 19103, E-mail: widdel{at}acnatsci.org. Linda J. McCuiston, Entomology Department, Rutgers University, 180 Jones Avenue, New Brunswick, NJ 08901. Wayne J. Crans, Entomology Department, Rutgers University, 180 Jones Avenue, New Brunswick, NJ 08901. Laura D. Kramer, Wad-sworth Center, New York State Department of Health, 5668 State Farm Rd., Slingerlands, NY 12159. Dina M. Fonseca, Academy of Natural Sciences, 1900 Ben Franklin Parkway, Philadelphia, PA 19103, Telephone: 215-299-1177, Fax: 215-299-1182, E-mail: fonseca{at}acnatsci.org.

Reprint requests: Dina M. Fonseca, Academy of Natural Sciences, 1900 Ben Franklin Parkway, Philadelphia, PA 19103, Telephone: 215-299-1177, Fax: 215-299-1182, E-mail: fonseca{at}acnatsci.org.