First Molecular Epidemiological Study of Cutaneous Leishmaniasis in Libya

Cutaneous leishmaniasis (CL) is caused by protozoan parasites of the genus Leishmania. The disease is characterized by the formation of chronic skin lesions followed by permanent scars and deformation of the infected area. It is distributed in many tropical and subtropical countries with more than 2 million cases every year. During the past few years CL has emerged as a major public health problem in Libya. So far, diagnosis was based on clinical symptoms and microscopic observation of parasites. Disease outbreaks were not investigated and the causative leishmanial species of CL were not identified so far. Our study indicates the presence of two coexisting species: Leishmania major and Leishmania tropica. These results are crucial in order to provide accurate treatment, precise prognosis and appropriate public health control measures. The recent armed conflict in Libya that ended with the Gadhafi regime collapse on October 2011 has affected all aspects of the life in the country. In this study we discussed multiple risk factors that could be associated with this conflict and present major challenges that should be considered by local and national health authorities for evaluating the CL burden and highlighting priority actions for disease control

First report of Leishmania (L) killicki Rioux, Lanotte & Pratlong, 1986 in Libya.

International audienc

Advantages and limits of real-time PCR assay and PCR-restriction fragment length polymorphism for the identification of cutaneous Leishmania species in Tunisia.

International audienceCutaneous 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

Mucosal leishmaniasis of the lip: an emerging clinical form in Tunisia

International audienceBackgroundMucosal leishmaniasis (ML), which mostly occurs in the New World, is mainly associated with Leishmania braziliensis. Primary lip ML is very rare in the Mediterranean basin and particulary in Tunisia despite the endemicity of both cutaneous and visceral leishmaniasis in this area. ObjectivesTo highlight a recent emergence of primary lip ML in Tunisia, to describe its epidemiological and clinical features and to identify the causative Leishmania species. MethodsEpidemiological, clinical and therapeutic data of 10 cases presenting a ML of the lip were collected. Diagnosis confirmation of leishmaniasis was obtained by microscopic examination of Leishmania parasites in Giemsa stained smears of the lesion sampling and in cutaneous biopsies. Polymerase chain reaction (PCR) detecting Leishmania DNA directly from dermal scraping was also performed for diagnosis and species identification. ResultsSeven men and three women with lip ML were diagnosed during the last 6years (2008-2013). The mean age was 29.7years. Clinical presentation was characterized by an infiltrated and ulcerated plaque leading to macrocheilitis involving the upper lip in eight cases and the lower lip in two cases. Mean diagnosis delay was 6.9months. PCR identified L. infantum in seven cases and L. major in two cases. Seven patients received intramuscular injections of meglumine antimoniate (MA) and three patients received both MA intralesional injections of MA and cryotherapy. A clinical remission was rapidly observed in all cases (on average in 2.2months). ConclusionsPrimary 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

Caractérisation comparative des trois formes de leishmaniose cutanée endémiques en Tunisie

International audienceBackground : The recent spread in the geographical distribution of the three forms of cutaneous leishmaniasis (CL) endemic in Tunisia has resulted in the coexistence of more than one species of Leishmania (L.) in some foci, rendering characterization on the basis of geographical criteria alone more difficult. The aim of the study was to establish clinical criteria associated with these noso-geographic forms, namely sporadic CL (SCL) due to L. infantum, zoonotic CL (ZCL) due to L. major and chronic CL (CCL) due to L. tropica.Patients and methods : One hundred and twelve patients with biologically confirmed CL were involved in the study. Leishmania species was systematically identified by iso-enzyme analysis and/or PCR-RFLP. Details of the number, the location, the morphological aspect and the month of outbreak of the lesions were noted for each patient.Results : SCL lesions appeared later than ZCL lesions (53.8% of cases appeared from December onwards vs. 23.6%, P < 0.001). ZCL lesions were often multiple (75%) and situated on the limbs (84.7%, P < 0.001), whereas SCL lesions were single (92.3%, P < 0.001) and located on the face (84.6%, P < 0.001). CCL lesions were also single (78.6%) and located on the face (71.4%). The classical ulcerous presentation with scabs was mainly observed in ZCL patients (69.4%) and the erythematous presentation was described more frequently in SCL patients (75%; P < 0.001).Conclusion : The number, site, morphological aspect and month of outbreak of lesions could be considered as useful criteria that help differentiate between the three noso-geographical forms of CL prevailing in Tunisia. Such characterization is useful for the individual management of patients and for optimizing the combat against the disease.Introduction : Les extensions récentes des aires de répartition des trois formes de leishmaniose cutanée (LC) endémiques en Tunisie font que désormais plus d’une espèce coexistent dans certains foyers, ce qui empêche la discrimination entre ces différentes formes à partir des seuls critères géographiques. L’objectif de ce travail est d’identifier certains critères cliniques qui seraient associés aux formes en question : la LC sporadique (LCS) due à Leishmania infantum, la LC zoonotique (LCZ) due à L. major et la LC chronique due à L. tropica.Patient et méthode : Cent douze cas de LC confirmés biologiquement ont fait partie de l’étude. L’espèce de leishmanie en cause a été systématiquement identifiée par électrophorèse des iso-enzymes et/ou PCR-RFLP. Étaient relevés systématiquement le nombre et la localisation des lésions, ainsi que leur aspect morphologique et le mois de leur apparition.Résultats : Les lésions de LCS semblent se déclarer plus tardivement que celles de LCZ (53,8 % à partir de décembre vs. 23,6 %, p < 0,001). Les lésions de LCZ sont souvent multiples (75 %) et localisées aux membres (84,7 %), alors que celles de LCS sont plutôt uniques (92,3 %, p < 0,001) et touchent le visage (84,6 %, p < 0,001). La LCC se présente également sous forme de lésions uniques (78,6 %) du visage (71,4 %). L’aspect ulcéro-croûteux dominait les présentations des lésions de LCZ (69,4 %) et l’aspect érythémateux infiltré celles de LCS (75 %, p < 0,001).Conclusion : En complément de la provenance géographique des cas, le nombre des lésions, leur localisation, leur aspect morphologique et le mois où elles surviennent facilitent la discrimination entre les trois formes de LC présentes en Tunisie, ce qui est utile pour la prise en charge des patients et l’adaptation des mesures de contrôle

Epidemiologic and Clinical Features of Cutaneous Leishmaniasis in Southeastern Tunisia

Species-specific diagnosis was performed in 66 patients with cutaneous leishmaniasis (CL) living in Tataouine focus in southeastern Tunisia. Leishmania DNA was extracted directly from dermal scrapings (n = 66) and from parasites obtained in culture (n = 12). Species were identified by using polymerase chain reaction–restriction fragment length polymorphism analysis for internal transcribed spacer region 1 and isoenzyme analysis. Leishmania tropica and L. major were identified in 31 (47%) and 35 (53%) cases respectively. Leishmania tropica CL cases were geographically scattered, and L. major CL cases were clustered. Lesions caused by L. tropica were mostly single (83.8%) and face-localized (55.8%), and lesions caused by L. major were multiple (57.1%; P < 0.001) and situated on limbs (83.7%; P < 0.001). For both species, most lesion onsets were reported during June–January. However, lesions that emerged during February–May were mainly caused by L. tropica (83.3%; P < 0.01). Moreover, the delay before seeking medical advice was higher for L. tropica infections than for L. major infections (P < 0.05)