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Development of a Molecular Tool for the Identification of Leishmania Reservoir Hosts by Blood Meal Analysis in the Insect Vectors

The transmission of parasites of the genus Leishmania involves a large diversity of mammalian reservoir hosts. However, many of these are yet to be identified, mainly in isolated biotopes such as the Amazonian rain forest. Furthermore, the trophic preferences of insect vectors have major epidemiologic implications. In this study, we developed a molecular tool for the identification of blood meals of phlebotomine sand flies. This assay is based on specific amplification and sequencing of the blood meal–derived single copy prepronociceptin (PNOC) gene, which is used as a target in phylogenetic studies of mammals. Sand flies were identified simultaneously with the blood-meal identification, using molecular analysis of a ribosomal locus. After a systematic assessment of the sensitivity and specificity of polymerase chain reaction amplification of the PNOC gene using human fed sand flies, the assay was tested on wild-caught sand flies. This work has important implications for the discovery of new Leishmania reservoir hosts and for a better understanding of complex parasite life cycles.

Received March 26, 2007. Accepted for publication August 20, 2007.

Acknowledgments: The authors thank Ghyslaine Serres and Samia Boussaa for technical assistance, Dick Ashford for critical reading of the manuscript, and three anonymous expert referees for constructive remarks.

Financial support: This work was supported by finances from the Tunisian Ministry of Scientific Research, Technology and Competences Development.

^* Address correspondence to Christophe Ravel, 163 rue Broussonet, Montpellier 34090, France. E-mail: christophe.ravel{at}univ-montp1.fr

Authors’ addresses: Najoua Haouas, Laboratory of Parasitology code 99UR/08-05, Department of Clinical Biology B, Faculty of Pharmacy, 1 Rue Avicenne 5000, Monastir, Tunisia, Telephone: 216-73-448-666. Fax: 216-73-461-830, E-mail: najoua.h{at}laposte.net. Bernard Pesson, Louis Pasteur University, Strasbourg I, Department of Parasitology, Faculty of Pharmacy, BP 60024, 74 route du Rhin, 67401 Illkirch Cedex, France, Telephone: 33-0-3-90-24-42-05, Fax: 33-0-3-90-24-42-86, E-mail: bernard.pesson{at}pharma.u-strasbg.fr. Raja Boudabous, Laboratory of Parasitology code 99UR/08-05, Department of Clinical Biology B, Faculty of Pharmacy, 1 rue Avicenne 5000, Monastir, Tunisia, Telephone: 216-73-448-666, Fax: 216-73-461-830, E-mail: rajaboudabous{at}yahoo.fr. Jean-Pierre Dedet, Department of Parasitology, French National Reference Centre on Leishmaniasis, 163 Rue Auguste Broussonet, 34090 Montpellier, France, Telephone: 33-0-4-67-63-27-51, Fax: 33-0-4-67-63-00-49, E-mail: parasito{at}univ-montp1.fr. Hamouda Babba, Laboratory of Parasitology code 99UR/08-05, Department of Clinical Biology B, Faculty of Pharmacy, 1 rue Avi-cenne 5000, Monastir, Tunisia, Telephone: 216-73-448-666, Fax: 216-73-461-830, E-mail: hamouda.babba{at}gnet.tn. Christophe Ravel, Department of Parasitology, French National Reference Centre on Leishmaniasis, 163 Rue Auguste Broussonet, 34090 Montpellier, France, Telephone: 33-467-630-049, Fax: 33-467-632-751, E-mail: christophe.ravel{at}univ-montp1.fr.

Reprint requests: C. Ravel, Department of Parasitology, French National Reference Centre on Leishmaniasis, Institute of Botanic, 163 Rue Auguste Broussonet, 34090 Montpellier, France. E-mail: christophe.ravel{at}univ-montp1.fr.