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Establishment of a Mass Screening Method of Sand Fly Vectors for Leishmania Infection by Molecular Biological Methods

Surveillance of the prevalence of Leishmania and its vector, sand fly species, in endemic and surrounding areas is important for prediction of the risk and expansion of leishmaniasis. In this study, a method for the mass screening of sand flies for Leishmania infection was established. This method was applied to 319 field-captured specimens, and 5 positive sand flies were detected. Sand fly species were identified by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) of the18S rRNA gene, and all the positive flies were Lu. hartmanni. Furthermore, cytochrome b (Cyt b) gene sequence analyses identified all the parasites as Endotrypanum species including a probable novel species. Because the method requires minimum effort and can process a large number of samples at once, it will be a powerful tool for studying the epidemiology of leishmaniasis.

Received February 13, 2007. Accepted for publication May 1, 2007.

Acknowledgments: We thank Roberto Sud for technical assistance throughout the study; director Dr. Luiggi Martini and the coordinator Dr. Ernesto Gutierrez (NIH and Trop. Med., Ecuador) for arrangement of our field activities; Drs. Teresa Flor, Jenny Rodriguez, and Yim-Yan Wong Chum (NIH and Trop. Med., Ecuador) for support at the laboratory and field phases of the study; Drs. Yuichi Yotsuyanagi and Kenji Ninomiya (Shimadzu Biotech, Tsukuba, Japan) for technical assistance on Ampdirect PCR methods; Dr. Jesus G. Valenzuela (NIH, Bethesda, MD) for providing sand flies experimentally infected with L. (L.) major; and The DNA Core facility of the Center for Gene Research, Yamaguchi University, for technical expertise.

Financial support: This study was supported by the Ministry of Education, Science, Culture and Sports of Japan (Grants 14256004, 18256004, and 18780230).

^* Address correspondence to Hirotomo Kato, Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan, E-mail: katoh{at}yamaguchi-u.ac.jp

Authors’ addresses: Hirotomo Kato, Yoshimi Terayama, and Hiroyuki Iwata, Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan. Hiroshi Uezato, Department of Dermatology, Faculty of Medicine, University of the Ryukyus, Nishihara, Okinawa 903-0215, Japan. Eduardo A. Gomez, Department of Epidemiology and Parasitology, National Institute of Health and Tropical Medicine, Guayaquil, Ecuador. Manuel Calvopiña and Yoshihisa Hashiguchi, Department of Parasitology, Kochi Medical School, Kochi University, Nankoku, Kochi 783-8505, Japan.

Reprint requests: Hirotomo Kato, Department of Veterinary Hygiene, Faculty of Agriculture, Yamaguchi University, 1677-1 Yoshida, Yamaguchi 753-8515, Japan, E-mail: katoh{at}yamaguchi-u.ac.jp.