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Menzel Bourguiba, Tunisia

Mouttaki T.,Center for Doctoral Studies in Health science | Mouttaki T.,Research Team on Cutaneous Leishmaniasis | Chiheb S.,University Hospital Ibn Rochd | Chiheb S.,Research Team on Cutaneous Leishmaniasis | And 4 more authors.
Parasites and Vectors | Year: 2014

Background: The diagnosis of cutaneous leishmaniasis (CL) might be difficult, in particular in endemic areas where different species of Leishmania can cause lesions of very similar appearance and where other skin diseases with similar clinical symptoms occur. Even today, the parasitological diagnosis of CL remains the gold standard and it is based on the direct identification of amastigotes in microscopy smears and/or culture of promastigotes from infected tissues. Although these techniques are highly specific, they are not sensitive enough. The objective of this study is to contribute to improving the diagnosis of CL and the identification of Leishmania species in Morocco by comparing three PCR-based assays applied directly on dermal samples.Methods: A total of 58 patients presenting with cutaneous lesions suggestive of CL were sampled for parasitological diagnosis by direct examination (DE), culture in NNN medium, two kinetoplast DNA (kDNA) PCRs (Lmj4/Uni21 and 13A/13B primers) and one rRNA gene internal transcribed spacer 1 (ITS1) PCR (LITSR/L5.8S primers). The techniques were statistically analyzed and compared.Results: According to our consensus positive, 44 out of 58 samples were true positives. The 13A/13B-PCR and ITS1-PCR showed the highest sensitivities (100%). Parasite microscopy and culture detected 43% and 29% of the true positives, respectively, while culture and microscopy together improved sensitivity to 52%. PCRs 13A/13B and ITS1 were associated to four and one false positives, respectively, while the other assays were 100% specific. Furthermore, the ITS1-PCR-RFLP assay clearly identified the Leishmania species for all the true positives (44/44), whereas Lmj4/Uni21-PCR identified 35/44 samples. The comparison between the Leishmania molecular characterizations and the expected species according to the national data from the Ministry of Health indicate 7 discrepant results.Conclusions: The PCR-based assays tested on our samples increased the speed and sensitivity of the diagnosis of CL compared to the conventional techniques. Furthermore, we showed that we can not base the species identification on the national data from the Ministry of Health. Finally, we suggest the use of PCR-ITS1-RFLP for diagnosis and simultaneous identification of the species in the Moroccan epidemiological context, but also in similar areas of the Mediterranean Basin. © 2014 Mouttaki et al.

Jaouad E.h.,Mohammed V University | M'hammed A.,Mohammed V University | Badr L.,Laboratory of Parasitology Mycology | Ali B.,Mohammed V University | Jamal T.,Mohammed V University
Research Journal of Pharmacy and Technology | Year: 2012

The treatment of hydatidosis is primarily surgical but can cause morbidity or mortality, hence the interest in many cases of medical treatment whose main representative is albendazole. The efficacy of this compound against Echinococcus granulosus cysts is proved, but limited by its poor bioavailability. New potential prodrugs were synthesized and their protoscolicidal activity was tested in vitro. Although hydrogen N°1 is important for the albendazole activity, some derivatives have kept an activity relatively important. © RJPT All right reserved.

Davoust B.,Animal Epidemiology Working Group of the Military Health Service | Davoust B.,Aix - Marseille University | Mary C.,Laboratory of Parasitology Mycology | Marie J.-L.,Animal Epidemiology Working Group of the Military Health Service
Journal of Wildlife Diseases | Year: 2014

The role of red foxes in the natural cycle of Leishmania infection is not well known. In the Var area, southeastern France, from 2006 to 2012, we conducted a longitudinal epidemiologic survey of foxes using quantitative PCR. Among 92 red foxes screened, prevalence of Leishmania infantum infection was 9%. Red foxes may be considered a bioindicator of parasite circulation in this biotope. © Wildlife Disease Association 2014.

Chaabane-Banaoues R.,University of Monastir | Oudni-M'rad M.,University of Monastir | M'rad S.,University of Monastir | Amani H.,University of Monastir | And 3 more authors.
Parasitology Research | Year: 2016

Cystic echinococcosis, due to Echinococcus granulosus sensu lato (s. l.), currently affects three million people, especially in low-income countries and results in high livestock production loss. DNA-based methods demonstrated genetic variability of E. granulosus s. l., and five species were recognized to belong to the complex, including E. granulosus sensu stricto (s.s) (genotypes G1–G3), Echinococcus equinus (genotype G4), Echinococcus ortleppi (genotype G5), Echinococcus canadensis (genotypes G6–G10), and the lion strain Echinococcus felidis. The characterization of Echinococcus species responsible for human and animal echinococcosis is crucial to adapt the preventive measures against this parasitic disease. The sequencing approach is the gold standard for genotyping assays. Unfortunately, developing countries do not often have access to these techniques. Based on in silico RFLP tools, we described an accurate PCR-RFLP method for Echinococcus spp. characterization. The double digestion with the HaeIII and HinfI restriction enzymes of the PCR product from nad1 gene (1071 bp) led to a clear discrimination between E. granulosus s. l. and most closely related species (Echinococcus shiquicus and Echinococcus multilocularis). Molecular procedures and phylogenetic analysis confirmed the efficiency and the reproducibility of this simple and fast PCR-RFLP method. This technique is proved useful for fresh/unfixed and FF-PET tissues and enables large-scale molecular epidemiological screening in developing countries. © 2016 Springer-Verlag Berlin Heidelberg

Vandenberg O.,Jules B | Vandenberg O.,Free University of Colombia | Robberecht F.,Saint Pierre University Hospital | Dauby N.,Jules B | And 5 more authors.
Pediatric Infectious Disease Journal | Year: 2012

Background: Cryptosporidium outbreaks in day-care centers (DCCs) occur commonly. However, controlling spread of infection in these settings is difficult, and data about effectiveness of different control strategies are sparse. In this study, a Cryptosporidium outbreak in a large DCC located in Brussels is described with evaluation of hygienic and therapeutic interventions. Methods: During a 3-week period, 43 of 130 children attending the DCC developed enteric symptoms. Stools from 122 children were examined for microbial pathogens. Of them, 38 (31%) were diagnosed with Cryptosporidium, 29 of them being symptomatic (76%) and 9 (24%) asymptomatic. Diagnosis was performed by microscopy, antigen tests, and real-time polymerase chain reaction. Strict infection control measures were implemented during the first week after the start of outbreak. After 4 weeks, 27/38 children (71%) were still symptomatic and Cryptosporidium positive. Because of persisting symptoms and fear of further spread of infection, all 27 children were treated with paromomycin. Two weeks later, 18 of 27 children were asymptomatic and were parasitologically negative. The remaining 9 children, still symptomatic and Cryptosporidium positive, were treated with nitazoxanide. Three weeks later, week 9 after the start of outbreak, all 38 children involved in the outbreak were asymptomatic and Cryptosporidium negative. Conclusions: Our study underscores the need to rule out Cryptosporidium etiology in a diarrheal outbreak in a DCC. Rapid implementation of infection control measures can most likely halt the spread of infection. The role of nitazoxanide to limit duration of shedding of oocysts deserves more attention for its use in outbreaks. © 2012 Lippincott Williams & Wilkins.

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