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      Genotypic profiles of Leishmania (Viannia) braziliensis strains from cutaneous leishmaniasis patients and their relationship with the response to meglumine antimoniate treatment: a pilot study Translated title: Profils génotypiques des souches de Leishmania (Viannia) braziliensis chez les patients atteints de leishmaniose cutanée et leur relation avec la réponse au traitement à l'antimoniate de méglumine : une étude pilote

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          Abstract

          Background: Forty-four strains isolated from a cohort of cutaneous leishmaniasis (CL) patients who did or did not respond to one course of treatment with meglumine antimoniate were investigated to explore genetic polymorphisms in parasite kinetoplast DNA minicircles. Leishmania (Viannia) braziliensis strains isolated from responder (R) and non-responder (NR) patients who acquired infection in Rio de Janeiro or in other Brazilian states were studied using low-stringency single-specific primer polymerase chain reaction (LSSP-PCR) to identify genetic polymorphisms. Results: Polymorphisms were observed in parasites recovered from patient lesions. No association was found between a specific genotype and R or NR patients. Phenetic analysis grouped the genotypes into three main clusters, with similarity indices varying from 0.72 to 1.00. Although no specific genotype association was detected, at least one group of L. (V.) braziliensis genotypes that circulates in Rio de Janeiro was discriminated in clusters I and III, showing phenotypes of good and poor responses to treatment, respectively. Cluster I comprised parasite profiles recovered from R patients from Rio de Janeiro and in cluster III, NR samples were prevalent. Cluster II comprised 24 isolates, with 21 from Rio de Janeiro and three from other states, equally distributed between R and NR patients. Additionally, we found that parasites sharing all common genetic characteristics acted differently in response to treatment. Conclusions: These results are of clinical-epidemiological importance since they demonstrate that populations of L. (V.) braziliensis that exhibit high levels of genetic similarity also display different phenotypes associated with meglumine antimoniate responses in cutaneous leishmaniasis patients.

          Translated abstract

          Contexte : Quarante-quatre souches isolées d'une cohorte de patients atteints de leishmaniose cutanée (CL) qui ont répondu ou non à un schéma de traitement avec de l'antimoniate de méglumine ont été étudiées pour explorer le polymorphisme génétique dans les minicerles d'ADN des kinétoplastes des parasites. Les souches de Leishmania (Viannia) braziliensis isolées des patients répondeurs (R) et non-répondeurs (NR) qui ont subi une infection à Rio de Janeiro ou dans d'autres états brésiliens ont été étudiées en utilisant une réaction en chaîne à la polymérase spécifique à faible stringence (LSSP-PCR) pour identifier le polymorphisme génétique. Résultats : Du polymorphisme a été observé chez les parasites récupérés dans les lésions de patients. Aucune association n'a été trouvée entre un génotype spécifique et les patients R ou NR. L'analyse phénétique a regroupé les génotypes en trois groupes principaux, avec des indices de similarité variant de 0,72 à 1,00. Bien qu'aucune association de génotype spécifique n'ait été détectée, au moins un groupe de génotypes de L. (V.) braziliensis qui circule à Rio de Janeiro a été discriminé dans les groupes I et III, montrant respectivement des phénotypes de bonnes et de mauvaises réponses au traitement. Le groupe I comprenait des profils de parasites récupérés chez des patients R de Rio de Janeiro et dans le groupe III, les échantillons NR étaient fréquents. Le groupe II comprenait 24 isolats, dont 21 de Rio de Janeiro et trois d'autres états, répartis de façon égale entre les patients R et NR. En outre, nous avons constaté que les parasites partageant toutes les caractéristiques génétiques communes agissaient différemment en réponse au traitement. Conclusions : Ces résultats sont d'importance clinico-épidémiologique, par la démonstration que les populations de L. (V.) braziliensis qui présentent des niveaux élevés de similarité génétique présentent également différents phénotypes associés aux patients atteints de leishmaniose cutanée et leur réponse à l'antimoniate de méglumine.

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          Most cited references 25

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          The molecular epidemiology and phylogeography of Trypanosoma cruzi and parallel research on Leishmania: looking back and to the future.

          Trypanosoma cruzi is the protozoan agent of Chagas disease, and the most important parasitic disease in Latin America. Protozoa of the genus Leishmania are global agents of visceral and cutaneous leishmaniasis, fatal and disfiguring diseases. In the 1970s multilocus enzyme electrophoresis demonstrated that T. cruzi is a heterogeneous complex. Six zymodemes were described, corresponding with currently recognized lineages, TcI and TcIIa-e--now defined by multiple genetic markers. Molecular epidemiology has substantially resolved the phylogeography and ecological niches of the T. cruzi lineages. Genetic hybridization has fundamentally influenced T. cruzi evolution and epidemiology of Chagas disease. Genetic exchange of T. cruzi in vitro involves fusion of diploids and genome erosion, producing aneuploid hybrids. Transgenic fluorescent clones are new tools to elucidate molecular genetics and phenotypic variation. We speculate that pericardial sequestration plays a role in pathogenesis. Multilocus sequence typing, microsatellites and, ultimately, comparative genomics are improving understanding of T. cruzi population genetics. Similarly, in Leishmania, genetic groups have been defined, including epidemiologically important hybrids; genetic exchange can occur in the sand fly vector. We describe the profound impact of this parallel research on genetic diversity of T. cruzi and Leishmania, in the context of epidemiology, taxonomy and disease control.
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            A general classification of New World Leishmania using numerical zymotaxonomy.

            More than 250 strains of Leishmania isolated from different localities and hosts in the New World were analyzed by enzyme electrophoresis, and their electromorphic profiles were compared with 19 reference strains representing most of the described species of this parasite. The 18 enzymic loci analyzed were very polymorphic, and the strains were classified into 44 zymodemes, each grouping strains with the same enzyme profiles. Each zymodeme was considered as an elementary taxon and the phenetic and phylogenetic relationships were determined by agglomerative hierarchical, ordination, and cladistic techniques. The different classification methods produced very similar results. The 44 zymodemes could be clustered into two groups, corresponding to the subgenera Leishmania and Viannia, by the numerical methods. The subgenus Viannia was shown to be monophyletic and could be further divided into species complexes representing L. braziliensis, L. naiffi, and L. guyanensis/L. panamensis/L. shawi, as well as some isolated taxa including L. lainsoni. The subgenus Leishmania, on the other hand, was polyphyletic, with New World isolates related to L. major clustered separately from the L. mexicana species complex. Most of the other zymodemes in this group represented independent taxa. The results confirm Viannia as a valid taxon but suggest that the status of the subgenus Leishmania should be further investigated. Leishmania braziliensis and L. naiffi were shown to be the most polymorphic species, while L. guyanensis, in spite of being the most common species found in this study, was remarkably homogeneous. The only variants were found south of the Amazon river. North of this river, the species was monomorphic.
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              Genetic polymorphism and molecular epidemiology of Leishmania (Viannia) braziliensis from different hosts and geographic areas in Brazil.

              Numerical zymotaxonomy and variability of the internal transcribed spacers (ITS) between the small and large subunits of the rRNA genes were used to examine strain variation and relationships in natural populations of Leishmania (Viannia) braziliensis. A total of 101 strains from distinct hosts and Brazilian geographic regions were assigned to 15 zymodemes clustered in two major genetic groups. The great number of isolates (48.5%) placed in zymodeme IOC/Z-27 were collected on the Atlantic coast. The high molecular diversity found in populations in the Amazon Basin was related to the great number of sandfly vector(s) in that region. The results of the restriction fragment length polymorphism analysis of the ITS depicted considerable intraspecific variation. Genotypic groups A, B, and C contained 39, 40, and 22 isolates, which were divided into 16, 10, and 15 genotypes, respectively. The genetic polymorphism observed demonstrates the degree of diversity of L. (V.) braziliensis strains from different regions where they are endemic. The results reinforce the clonal theory for Leishmania parasites showing the genetic diversity of this pathogen and an association of L. (V.) braziliensis genotypes with specific transmission cycles, probably reflecting an adaptation of different clones to the vector species involved.
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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2017
                29 September 2017
                : 24
                : ( publisher-idID: parasite/2017/01 )
                Affiliations
                [1 ] Laboratório de Pesquisa Clínica e Vigilância em Leishmanioses, Instituto Nacional de Infectologia Evandro Chagas INI/FIOCRUZ, Manguinhos, Rio de Janeiro Brazil
                [2 ] Departamento de Otorrinolaringologia e Oftalmologia, Faculdade de Medicina, UFRJ, Hospital Universitário Clementino Fraga Filho, Cidade Universitária - Ilha do Fundão, Rio de Janeiro Brazil
                Author notes
                [* ]Corresponding author: thaligagini@ 123456gmail.com
                Article
                parasite170060 10.1051/parasite/2017035
                10.1051/parasite/2017035
                5621350
                28959938
                © T. Gagini et al., published by EDP Sciences, 2017

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                Page count
                Figures: 3, Tables: 3, Equations: 0, References: 30, Pages: 11
                Categories
                Research Article

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