31 research outputs found

    Phylogenetic Analysis of the Oriental-Palearctic-Afrotropical Members of Anopheles (Culicidae: Diptera) Based on Nuclear rDNA and Mitochondrial DNA Characteristics

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    The phylogenetic relationships of Anopheles spp. at the junction of Oriental, Palearctic, and Afrotropical regions in the Iranian plateau were investigated using molecular markers. A 711-bp mitochondrial DNA cytochrome oxidase C subunit I (COI) fragment and the entire second internal transcribed spacer (ITS2) region (286-576 bp) of the nuclear ribosomal DNA (rDNA-ITS2) were sequenced from 14 and 28 taxa, respectively. The analyses included 12 species within Anopheles and 4 within the Myzorhynchus Series of the subgenus Anopheles, 8 within Neocellia, 6 within Myzomyia, 3 within Paramyzomyia, and 1 within the Pyretophorus Series of the subgenus Cellia. The congruent tree topologies of both molecular markers strongly supported monophyly of subgenera Anopheles and Phylogenetic trees constructed on the basis of IT52 sequences could accurately categorize all of the series according to the classical taxonomy but could not distinguish Pyretophorus (Anopheles subpictus) from Paramyzomyia Series. Although sequence data of the COI region were available for only 14 species, the inferred trees revealed good classification among the series but could not show the monophyletic relationship of Cellia spp. Except for a few cases, the tree inferred from ITS2 sequences revealed the best classification for the species studied. The molecular data could significantly improve our understanding of the phylogenetic position of the taxa

    Frequency and Molecular Identification of Leishmania Parasites in Smears Prepared from Skin Lesions of Patients Referred to Health Centers of Ilam Province by Digestion of the rDNA-ITS1 Gene

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    Objective: Ilam is a border province and a high risk zone for zoonotic cutaneous leishmaniasis (ZCL). Identification of Leishmania parasite species in clinical infections is a prerequisite for planning appropriate control measures. This study investigates the demographic characteristics of patients and molecular epidemiology of Leishmania parasites in the skin lesions of patients from Ilam Province. Methods: A total of 106 cases of suspected cutaneous leishmaniasis were detected passively and microscopic slides prepared from their active skin lesions. We randomly selected 50 slides. A fragment of the rDNA-ITS1 gene was amplified after which the PCR products digested with HaeIII restriction enzyme. There were 18 samples sequenced and their phylogenetic relationships compared with sequences retrieved from GenBank. Results: Leishmania amastigotes were detected in 100 slides. The highest and lowest distribution of cases was from the Moosian and Dehloran districts, respectively. There were 68.9% males and 31.1% of cases were women. The RFLP pattern of all samples was similar to Leishmania major. Phylogenetic relationships displayed great similarity between our sequences and those of Leishmania major parasites from sandflies trapped in Ilam and South Khorasan Provinces and human hosts from Esfarayen, Mahshahr and Afghanistan plus Leishmania mexicana of Venezuelan origin classified together in the same clade. Conclusion: Due to homogeneous morphology, problems associated with the cultivation of Leishmania and the two-step molecular identification process, the rDNA-ITS1-RFLP method has gained considerable attention in recent years. This method could be used as a very sensitive, simple, rapid and inexpensive method to detect Leishmania parasites in a variety of clinical and non-clinical samples

    First report on the shortest CPB peptide chain in the Leishmania donovani complex and bioinformatical interpretations in relation with this mutation

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    Background and Objective: Visceral Leishmaniasis or Kala-azar is an important infectious disease in northwestern Iran. Members of the Leishmania donovani complex, L. donovani and L.infantum, are the two main parasites causing the disease in the world. In this study immunophenotype characters such as N-glycosylation, and T-cell and B-Cell epitopes of CPB gene was evaluated in the Leishmania parasites isolated from sandflies of the region. Materials and Methods: Partial of the CPB (702-741 bp) of Leishmania parasites was amplified and sequenced and used for bioinformatics analysis such as test three dimensional (3D) protein structure, N-glycosylation, and T-cell and B-Cell epitopes. Results: In 7 out of 3477 sand flies there was a positive PCR test for systeine protease B, gene. Also according to findings of this study, both agents of kala-azar L. donovani and L. infantum were bing transfered by sand flies of Phlbtomus perfiliewi transcaucasicus in North West Iran. DNA analysis of the CPB gene showed a cytosine insertion at 5’ end of the proofreading frame of the gene resulted in a stop codon (TGA) seven AA further down and hence translation is halted. This caused a short amino acid chain with only 76 AA much shorter than normal CPB peptide with 234-247 AA. This mutation has not been found in the L.infantum strain resulted in a normal CPB peptide. AA analysis showed no N-glycosylation site, T-cell and B-Cell epitopes on the short peptide of the L. donovani strain. Conclusion: This is the shortest CPB peptide chain reported for the L. donovani complex in the literature. This short peptide could have an effect on host-parasite and vector-parasite interactions. Since the CPBs genes have important implication on host–parasite and play key roles in infection and expression of the disease, further studies on the L. donovani parasite and its diminutive peptide necessitate to improve our understanding about the epidemiology of visceral leishmaniasis in Iran

    Molecular detection of Leishmania

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