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Background Identification of vectors is of prime importance in the field of medical entomology for both operational and research purposes. An external quality assessment of mosquito identification capacities was carried out within the MediLabSecure Network, which is composed of laboratories located in 19 countries close to the European Union around the Mediterranean and Black seas. Methods A set of blind samples consisting of 7 or 8 adult mosquitoes and 4 larvae was given to each participant laboratory. In all, 138 adult mosquitoes and 76 larvae of different species were distributed for genus and species identification. Results All identifications were exclusively morphology based. Overall, 81% of identifications were correct at the genus level, 64% at the species level. The results were highly varied among the 19 participating laboratories. The levels of correct identifications were: 100% (three laboratories), 90–95% (four laboratories), 50–75% (six laboratories) and < 50% (six laboratories). Conclusions This evaluation showed the need to maintain efforts in capacity building and quality control in the field of medical entomology and, more specifically, in the morphological identification of the Culicidae. Electronic supplementary material The online version of this article (10.1186/s13071-018-3127-7) contains supplementary material, which is available to authorized users.

Visceral Leishmaniasis (VL) disease, endemic to Algeria, is a major public health problem for children from rural areas and suffering from malnutrition, this disease currently tends to decrease. However, we found a constant progression in adults over the last twenty years. Indeed, 64 cases of VL in adults were recorded between 1991 and 2010 in Kabylia (Tizi-Ouzou, Bejaia and Bouira) with 18 cases with VL/HIV co-infections. Co-infected men is predominant with 15 cases, a sex ratio of 5 and an average age 35.57 +/- 2.3 years. Typical clinical signs, namely; fever, splenomegaly and hepatomegaly are observed only in 22.2% of the cases; absence of fever (n=4), splenomegaly (n=2), two (n=2) and presence lymphadenopathy (n=5). We also noted the presence of risk factors such as the advanced age of patients and late diagnosis of visceral leishmaniasis usually associated with the advanced stages of HIV infection (III, IV) in mostly multi-infected patients. We also identified five (5) cases of shingles and four (4) cases of digestive candidacies who were the most opportunistic infections observed in our series. Pancytopenia, observed in 2/3 of the cases has been the most consistent biological sign and contributing in the evocation of the VL as the clinical presentation was atypical. The confirmation, by direct examination of bone marrow smears by identifying amastigotes, is observed in all HIV patients (PVIH). Western blot and PCR techniques have allowed better monitoring of patients. In 5 cases, the VL revealed the infection of HIV. The care was difficult because of the numerous observed relapses. The unavailability of Amphotericin B, inherent in different factors, compelled us, in some situations, to use the Glucantime as first-line treatment (n=4). Prophylactic treatment based on Pentamidine (n=2), Glucantime (n=2) and amphotericin B (n=12). The evolution was marked by 5 deaths during the first days of treatment anti-Leishmanial and one after several relapses were observed as well as a resistance perhaps to Glucantime resulting in splenectomy.

In Algeria, Leishmania infantum, Leishmania major, and Leishmania killicki (Leishmania tropica) are responsible for cutaneous leishmaniosis. We established a murine model of L. killicki infection to investigate its infective capacity, some immunophysiopathological aspects, and its suitability for pharmacological purposes. Following the injection of L. major or L. killicki metacyclic promastigotes in the ear dermis of BALB/c mice, the course of infection was followed. The infection with L. killicki caused slower lesion formation than with L. major. The presence of L. killicki or L. major DNA and parasites was detected in the ear dermis and in lymph nodes, spleen, and liver. Lesions induced by L. killicki were nonulcerative in their aspect, whereas those caused by L. major were highly ulcerative and necrotic, which matches well with the lesion phenotype reported in humans for L. killicki and L. major, respectively. The treatment of L. killicki lesions by injection of Glucantime (R) significantly reduced the lesion thickness and parasite burden. Ear dermal injection of BALB/c mice constitutes a model to study lesions physiopathology caused by L. killicki and presents interest for in vivo screening of new compounds against this pathogen, emerging in Algeria.

International audience; AIM: Three forms of cutaneous leishmaniasis (CL) are endemic in Tunisia. The identification of the causative species is useful to complete epidemiological data and to manage the cases. The aim of this study is to assess PCR-RFLP technique in the identification of Leishmania species responsible of CL in Tunisia and to compare the results of this technique to those of isoenzyme analysis. PATIENTS AND METHODS: Sixty-one CL lesions were sampled. Dermal samples were tested by culture on NNN medium and analyzed by PCR-RFLP assay targeting the ITS1 region of ribosomal DNA. Species identification was performed by both iso-enzymatic typing for positive cultures and analysis of restriction profiles after enzymatic digestion by HaeIII of the obtained amplicons. RESULTS: Thirty-eight (62%) samples were positive by culture. The iso-enzymatic typing of 32 isolates identified 3 L. infantum, 23 L. major MON-25 and 6 L. tropica MON-8. Sixty samples were positive by PCR. The PCR-RFLP digestion profiles of the 56 PCR products identified 12 L. infantum, 38 L. major and 6 L. tropica. The results of both techniques were concordant in the 32 strains identified by both techniques. Species identification correlated with the geographical distribution of CL forms endemic in Tunisia. CONCLUSION: Results of PCR-RFLP revealed highly concordant with those of isoenzyme electrophoresis. Thanks to its simplicity, rapidity and ability to be performed directly on biological samples, this technique appears as an interesting alternative for the identification of Leishmania strains responsible of CL in Tunisia.

This study presents for the first time the identification of Leishmania infantum zymodeme MON-80 in 6 dogs stemming from Algeria and Tunisia using the method of comparative electrophoresis migration of the parasite iso-enzymes. As this enzymatic variant has been already described in humans with cutaneous and visceral forms of leishmaniasis in south Europe and in the Maghreb (Algeria and Tunisia). these observations reinforce the role of the dog as the principal reservoir host for Leishmania infantum complex., Cette étude présente l’identification, pour la première fois, de Leishmania infantum zymodème MON-80 chez 6 chiens provenant d’Algérie et deTunisie par la méthode de migration électrophorétique comparatives des iso-enzymes parasitaires. Ce variant enzymatique ayant déjà été signalé chez l’homme lors de leishmanioses viscérales et cutanées en Europe du sud et au Maghreb (Algérie et Tunisie), ces résultats viennent renforcer le rôle du chien comme réservoir principal du complexe Leishmania infantum.

This study presents for the first time the identification of Leishmania infantum zymodeme MON-80 in 6 dogs stemming from Algeria and Tunisia using the method of comparative electrophoresis migration of the parasite iso-enzymes. As this enzymatic variant has been already described in humans with cutaneous and visceral forms of leishmaniasis in south Europe and in the Maghreb (Algeria and Tunisia). these observations reinforce the role of the dog as the principal reservoir host for Leishmania infantum complex., Cette étude présente l’identification, pour la première fois, de Leishmania infantum zymodème MON-80 chez 6 chiens provenant d’Algérie et deTunisie par la méthode de migration électrophorétique comparatives des iso-enzymes parasitaires. Ce variant enzymatique ayant déjà été signalé chez l’homme lors de leishmanioses viscérales et cutanées en Europe du sud et au Maghreb (Algérie et Tunisie), ces résultats viennent renforcer le rôle du chien comme réservoir principal du complexe Leishmania infantum.

International audience; The epidemiological situation of leishmaniasis in Tunisia is characterised by the co-existence in a very limited territory (165,000 km2, Sahara included), of 4 clinical forms: the infantile visceral leishmaniasis (VL) and 3 cutaneous leishmaniasis (CL) forms (sporadic, zoonotic and chronic). In addition to the useful epidemiological data, identification of the causative parasitic species is essential to determine the geographic distribution of each form and to select appropriate therapeutic procedure and suitable control measures. 226 Leishmania isolates, 135 human's coming from 59 VL cases and 76 CL cases and 91 canine's were identified by the isoenzyme electrophoresis reference technique. Results confirm the endemicity of the 4 forms mentioned above. The sporadic CL, confined to the North of the country is principally caused by L. infantum MON-24 (72.2%). VL which has reached the southern ridge in the central area of Tunisia, in the governorate of Kairouan (36 typed isolates), presents an unusual high proportion of L. infantum MON-24. In fact, this zymodeme, rather dermotropic is responsible for 47.2% of the cases vs 13% in the other regions of the country where L. infantum MON-1 remains predominant with 78.3% of typed isolates, the difference being statistically significant (P < 0.01). A third zymodeme, L. infantum MON-80 is sporadically pointed out during VL or in sporadic CL. Despite the high number of canine isolates (n=91) coming from 6 governorates, only the zymodeme L. infantum MON-1 was identified, letting hypothetic the reservoir of the 2 other zymodemes of the species identified in humans. Those absences may be related to cross infections, with a low sensitivity to L. infantum MON-24 leading to a selection of MON-1 at the time of culture passages. Hence it is important to develop molecular tools of direct identification on initial biological samples without going through cultures. Zoonotic CL remains the predominant cutaneous form in the central and southern area of Tunisia. However L. killicki, agent of the chronic CL, is confirming its presence out of its original focus of Tataouine in the southern-east of the country in both zoonotic CL and VL areas.

Epidemiological, clinical and parasitological data concerning canine leishmaniasis were collected in two Tunisian populations of dogs, different in breed and life style: 23 rural dogs and 26 dogs of European breeds. All were symptomatic and lived in the north of Tunisia where human visceral and cutaneous leishmaniasis caused by Leishmania infantum are endemic. Leishmaniasis has been confirmed in all dogs by serology or/and by parasitic identification. Significant differences concerning age and symptoms suggest a higher susceptibility to infection in European imported breeds. In fact individuals of this population were significantly younger; 81% were less than 5 years old whereas 57% of the autochthonous rural dogs were more than 5 years old with 31% of them being older than 9 (p = 0.014). In spite of their young age, 75% of imported breeds presented an affection of their general state with more frequent cutaneous symptoms than the rural dogs (96% versus 69%, p = 0.02). Isoenzyme typing of 31 strains, obtained from the two populations, from different sites (blood, lymph nodes, spleen) has only identified the zymodeme Leishmania infantum MON-1. This stresses the need of more investigations to determine reservoirs of the other enzymatic variants identified in humans in Tunisia and Mediterranean basin.

Iso-enzymatic characterization of 19 Leishmania strains isolated from Tunisian dogs revealed that all correspond to Leishmania infantum MON-1. This confirms the role of dog as a reservoir of the commonest zymodeme responsible for human visceral leishmaniasis in the Mediterranean basin. However, and in spite of the high number of identifications, many zymodemes, mainly the dermotropic ones as Leishmania infantum MON-24, frequently reported in humans, had never been isolated from dogs. The study of their cultural characteristics may contribute to explain the particular iso-enzymatic profile of dogs strains., Dix-neuf souches de leishmanies, isolées chez des chiens originaires de différentes régions de la Tunisie, ont été typées par électrophorèse des iso-enzymes. Toutes se sont révélé correspondre à Leishmania infantum MON-1, confirmant la prédominance de ce zymodème chez cet animal. L’absence, aussi bien dans cette série que dans celles d’autres pays méditerranéens, de certains zymodèmes dermotropes, fréquemment rencontrés chez l’homme, pose le problème de leur réservoir. La multiplication des isolements chez les hôtes potentiels et l’étude de la cinétique en culture des différents zymodèmes pourraient apporter d’éventuelles réponses.

The microscopic study of the dermal smears of 62 cases of cutaneous leishmaniose, 27 infected by Leishmania (L.) infantum and 35 by L. major, showed that the amastigotes of L. infantum are meaningfully smaller (p \textless 0.001). This criteria is a simple pary alternative to distinguish these 2 species which have completely different epidemiology, recovery delay and prophylactic dispositions., L’étude microscopique des frottis dermiques de 62 sujets atteints de leishmaniose cutanée, 27 infectés par Leishmania (L.) infantum et 35 par L. major, a montré que les formes amastigotes de L. infantum sont significativement plus petites (p

The isoenzymatic typing of 16 stocks of Leishmania, isolated from Tunisian visceral leishmaniasis (VL) cases, revealed that all strains belonged to Leishmania (L.) infantum species. Although zymodeme MON-1 was predictably the most frequent (9 cases), it came as a surprise that L. infantum MON-24 was responsible of about third of cases. This latter zymodeme, while previously reported in Tunisia, Algeria and Spain, was assumed to be a dermotropic and the few cases of VL that it caused occurred always in HIV infected patients. L. infantum MON-80, occasionally reported during both cutaneous and VL of immunocompetent infants was identified in 2 patients. This report confirms that in addition to the more common L. infantum MON-1, zymodeme MON-24 has a substantial role in generating VL in immunocompetent infants in Tunisia., Le typage iso-enzymatique de 16 souches de leishmanies isolées chez des enfants tunisiens atteints de leishmaniose viscérale (LV) a révélé qu'en plus du classique Leishmania (L.) infantum MON-1, identifié chez 9 patients, le zymodème L. infantum MON-24 a été trouvé dans 5 cas. Bien que ce zymodème ait déjà été incriminé dans la genèse de quelques cas de LV infantile en Algérie, en Tunisie et en Espagne, une proportion aussi élevée, 31,2%, était inattendue. En effet, L. infantum MON-24 a été jusque-là considéré comme plutôt dermotrope et les quelques cas de LV qui lui sont rapportés sont survenus le plus souvent chez des individus immunodéprimés infectés par le VIH.Par ailleurs, L. infantum MON-80, déjà signalé au cours de formes aussi bien cutanées que viscérales, a été identifié chez 2 patients.

The isoenzymatic typing of 16 stocks of Leishmania, isolated from Tunisian visceral leishmaniasis (VL) cases, revealed that all strains belonged to Leishmania (L.) infantum species. Although zymodeme MON-1 was predictably the most frequent (9 cases), it came as a surprise that L. infantum MON-24 was responsible of about third of cases. This latter zymodeme, while previously reported in Tunisia, Algeria and Spain, was assumed to be a dermotropic and the few cases of VL that it caused occurred always in HIV infected patients. L. infantum MON-80, occasionally reported during both cutaneous and VL of immunocompetent infants was identified in 2 patients. This report confirms that in addition to the more common L. infantum MON-1, zymodeme MON-24 has a substantial role in generating VL in immunocompetent infants in Tunisia., Le typage iso-enzymatique de 16 souches de leishmanies isolées chez des enfants tunisiens atteints de leishmaniose viscérale (LV) a révélé qu'en plus du classique Leishmania (L.) infantum MON-1, identifié chez 9 patients, le zymodème L. infantum MON-24 a été trouvé dans 5 cas. Bien que ce zymodème ait déjà été incriminé dans la genèse de quelques cas de LV infantile en Algérie, en Tunisie et en Espagne, une proportion aussi élevée, 31,2%, était inattendue. En effet, L. infantum MON-24 a été jusque-là considéré comme plutôt dermotrope et les quelques cas de LV qui lui sont rapportés sont survenus le plus souvent chez des individus immunodéprimés infectés par le VIH.Par ailleurs, L. infantum MON-80, déjà signalé au cours de formes aussi bien cutanées que viscérales, a été identifié chez 2 patients.

This study refers to 23 patients presenting with the sporadic forms of cutaneous leishmaniasis encountered in northern most humid parts of Tunisia. Culture inoculation for parasitic isolation was processed using two media: the classical NNN and a rabbit serum based medium (SLC). Cultures were positive in 17 cases with SLC medium and 13 cases with NNN medium. Eight isolates were typed using 15 isoenzymes systems. Six isolates were identified as Leishmania infantum MON-24 which confirms the crucial role of this zymodeme in causing this form of cutaneous leishmaniasis. The other two isolates were identified as Leishmania infantum MON-1, which is the principal agent of visceral leishmaniasis in the Mediterranean area., Vingt-trois prélèvements provenant de sujets atteints de leishmaniose cutanée sporadique du nord tunisien (LCS) ont été mis en culture sur le classique milieu NNN et le milieu sérum de lapin coagulé (SLC). 17 cultures s'étaient révélées positives avec le milieu SLC contre seulement 13 avec le milieu NNN. Huit parmi les 17 isolats de leishmanies obtenus ont été typés par une méthode utilisant 15 systèmes iso-enzymatiques. Six souches ont été identifiées comme étant Leishmania infantum MON-24, confirmant le rôle prédominant de ce zymodème dans la genèse de la LCS. Les deux autres souches ont été identifiées comme étant Leishmania infantum MON-1, agent classique de la leishmaniose viscérale dans le bassin méditerranéen.

This study refers to 23 patients presenting with the sporadic forms of cutaneous leishmaniasis encountered in northern most humid parts of Tunisia. Culture inoculation for parasitic isolation was processed using two media: the classical NNN and a rabbit serum based medium (SLC). Cultures were positive in 17 cases with SLC medium and 13 cases with NNN medium. Eight isolates were typed using 15 isoenzymes systems. Six isolates were identified as Leishmania infantum MON-24 which confirms the crucial role of this zymodeme in causing this form of cutaneous leishmaniasis. The other two isolates were identified as Leishmania infantum MON-1, which is the principal agent of visceral leishmaniasis in the Mediterranean area., Vingt-trois prélèvements provenant de sujets atteints de leishmaniose cutanée sporadique du nord tunisien (LCS) ont été mis en culture sur le classique milieu NNN et le milieu sérum de lapin coagulé (SLC). 17 cultures s'étaient révélées positives avec le milieu SLC contre seulement 13 avec le milieu NNN. Huit parmi les 17 isolats de leishmanies obtenus ont été typés par une méthode utilisant 15 systèmes iso-enzymatiques. Six souches ont été identifiées comme étant Leishmania infantum MON-24, confirmant le rôle prédominant de ce zymodème dans la genèse de la LCS. Les deux autres souches ont été identifiées comme étant Leishmania infantum MON-1, agent classique de la leishmaniose viscérale dans le bassin méditerranéen.

Archives des bulletins de la Société de Pathologie Exotique : http://www.pathexo.fr/documents/articles-bull/T92-1-1997.pdf; International audience; A definitive confirmation of the presence in Tunisia of the rare zymodem MON-80 of Leishmania infantum, as a causative agent of infantile visceral leishmaniasis, is reported.