Cutaneous leishmaniasis (CL), a public health problem in Tunisia, is associated to three species: Leishmania (L.) infantum, L. major and L. killicki. Accurate and sensitive procedures for the diagnostic of Leishmania infection and for species identification are required to enable adequate treatment and appropriate control measures. Several PCR-methods are applied for the diagnosis and the identification of Leishmania parasites such as PCR-restriction fragment length polymorphism (PCR-RFLP), DNA sequencing, hybridization probes and real-time PCR (RT-PCR). In this study, PCR-RFLP and RT-PCR were performed on skin scrapings from 27 patients with confirmed CL by microscopic examination, in order to compare their usefulness and efficiency for identification of Leishmania species in routine diagnostic laboratories. Identification of Leishmania species was successfully achieved in 96.3% and 81.5% respectively. Agreement between using internal transcribed spacer 1 (ITS1)-PCR-RFLP and kDNA-RT-PCR assays was 70% (19/27). Characterization problems using RT-PCR were mainly due to the difficulties in analyzing the melting temperatures. ITS1-PCR-RFLP and kDNA-RT-PCR presented an interesting alternative to conventional methods for the identification of Leishmania parasites from clinical samples. Both PCR assays can be used in a routine diagnostic, however, further prospective studies including largest sampling, are required to determine their performances in a routine use.

Abstract. Blastocystis sp. is currently the most common eukaryotic parasite found in humans. Despite its potential public health impact, epidemiological data regarding its prevalence and molecular subtype (ST) distribution in Maghreb are rarely reported. Therefore, the aim of this study was to determine the prevalence of the parasite in a cohort of healthy food handler Tunisian individuals and to acquire the first molecular data regarding the distribution of Blastocystis sp. STs in this country. Therefore, 524 fecal samples were collected, and 68 of them (13%) were identified as positive for the parasite by direct-light microscopy of smears. Seventeen samples of 100 negative by microscopy were also shown to be positive by real-time quantitative polymerase chain reaction. Among all the positive samples, 61 Blastocystis isolates were subtyped using partial small subunit ribosomal RNA gene analysis. ST3 was the most abundant (51%) followed by ST1 (30%), ST2 (16%), and ST4 and ST7 (both 1.6%).Blastocystis sp. is a parasite inhabiting the gastrointestinal tract of humans and a wide range of animals. This singlecelled eukaryote is currently the most prevalent protozoa found in human fecal samples with a worldwide distribution.

Primary lip ML is emerging in Tunisia. Macrocheilitis of the upper lip is the main clinical presentation. PCR revealed more sensitive than direct examination in the diagnosis of such form (P < 0.01). Leishmania infantum was the most identified species (7 cases) while L major was involved in only two lesions. A benign local evolution and a rapid recovery were observed in all cases after MA treatment.

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