title: First detection of Leishmania major DNA in Sergentomyia (Sintonius) clydei (Sinton, 1928, Psychodidae: Phlebotominae), from an outbreak area of cutaneous leishmaniasis in Tunisia.
abstract: In recent years there has been growing interest in Sergentomyia species. Their role in the spread of mammalian leishmaniasis appears repeatedly in the literature and the possibility of its implication in Leishmania transmission to humans remains controversial. Sergentomyia (Sintonius) clydei is one of several cryptic species sharing therefore common morphologic criteria with others species of the subgenera Sintonius. Little is known about this specie in Tunisia. We sampled and identified different specimens including four specimens of S. clydei collected from Sidi Bouzid and Kairouan areas (center of Tunisia) using morphological tools. Male Sergentomyia clydei and Sergentomyia christophersi are known to share several morphological characters and can be mistaken for. Consequently we took advantage of 5 male S. christophersi available in our collection (Tataouin, South of Tunisia). In our study morphological tools were completed by molecular study of cytochrome b gene to identify S. clydei. For the detection of Leishmania spp. that might infect our specimens, Leishmania DNA was analyzed by amplification of kinetoplast minicircle DNA using real-time PCR and nested-PCR. Obtained result was confirmed by restriction analysis of the amplified ribosomal internal transcribed spacer 1 (ITS1). We provide in our study, the first molecular identification of S. clydei, in Tunisia. Our Neighbor Joining tree based on mitochondrial cytochrome b gene shows two different clusters. The first includes the Tunisians S. clydei and other specimens from Africa, Middle East and the Arabic peninsula, and the second cluster containing the specimens from Seychelle. Unexpectedly, we also demonstrate the infection of one anthropophilic female S. clydei by Leishmania major DNA. This finding shows that more attention should be paid when identifying parasites by molecular tools within sandfly vector.
authors.affiliation: Laboratoire de Parasitologie Médiale, Biotechnologies et Biomolécules (LR11IPT06), Institut Pasteur de Tunis, 13, Place Pasteur-BP74, 1002 Tunis, Tunisia; Faculté des Sciences de Bizerte, Université de Carthage, Tunis, Tunisia.
authors.forename: Chiraz
authors.initials: C
authors.lastname: Ayari
authors.affiliation: Laboratoire de Parasitologie Médiale, Biotechnologies et Biomolécules (LR11IPT06), Institut Pasteur de Tunis, 13, Place Pasteur-BP74, 1002 Tunis, Tunisia.
authors.forename: Souad
authors.initials: S
authors.lastname: Ben Othman
authors.affiliation: Laboratoire d'Epidémiologie Moléculaire et Pathologie Expérimentale Appliquée aux Maladies Infectieuses (LR11IPT04), Institut Pasteur de Tunis, 13, Place Pasteur-BP74, 1002 Tunis, Tunisia.
authors.forename: Jomaa
authors.initials: J
authors.lastname: Chemkhi
authors.affiliation: Laboratoire de Parasitologie Médiale, Biotechnologies et Biomolécules (LR11IPT06), Institut Pasteur de Tunis, 13, Place Pasteur-BP74, 1002 Tunis, Tunisia.
authors.forename: Ahmed
authors.initials: A
authors.lastname: Tabbabi
authors.affiliation: Laboratoire de Parasitologie, Faculté de Pharmacie, Université de Barcelone, Av. Joan XXIII s/n, 08028 Barcelone, Spain.
authors.forename: Roser
authors.initials: R
authors.lastname: Fisa
authors.affiliation: Laboratoire de Transmission, Contrôle et Immunologie des Infections (LR11IPT02), Institut Pasteur de Tunis, 13, Place Pasteur-BP74, 1002 Tunis, Tunisia.
authors.forename: Afif
authors.initials: A
authors.lastname: Ben Salah
authors.affiliation: Laboratoire de Parasitologie Médiale, Biotechnologies et Biomolécules (LR11IPT06), Institut Pasteur de Tunis, 13, Place Pasteur-BP74, 1002 Tunis, Tunisia. Electronic address: souha.benabderrazak@pasteur.rns.tn.
authors.forename: Souha
authors.initials: S
authors.lastname: BenAbderrazak
mesh_terms: D000818:Animals; D045303:Cytochromes b / Q000235:genetics; D018105:DNA, Kinetoplast; D016054:DNA, Protozoan; D004196:Disease Outbreaks; D005260:Female; D006801:Humans; D018320:Leishmania major / Q000235:genetics*; D016773:Leishmaniasis, Cutaneous / Q000453:epidemiology* / Q000469:parasitology* / Q000635:transmission; D008297:Male; D010802:Phylogeny; D011576:Psychodidae / Q000033:anatomy & histology / Q000145:classification / Q000235:genetics / Q000469:parasitology*; D014416:Tunisia / Q000453:epidemiology
chemical_list: D018105:DNA, Kinetoplast; D016054:DNA, Protozoan; D045303:Cytochromes b
keywords: Cytochrome b; Kinetoplast minicircle DNA; Leishmania major; Ribosomal internal transcribed spacer 1; Sergentomyia clydei; Tunisia