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      First report of Leishmania ( Viannia) lindenbergi causing tegumentary leishmaniasis in the Brazilian western Amazon region Translated title: Premier signalement de Leishmania (Viannia) lindenbergi causant une leishmaniose tégumentaire dans la région amazonienne de l’ouest du Brésil

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          Abstract

          Tegumentary Leishmaniasis (TL) in the Brazilian Amazon region is associated with several Leishmania species. In this report, we describe two cases of TL related to Leishmania lindenbergi occurring in different locations of Rondônia state. After clinical diagnosis, lesion samples were collected for parasitological diagnoses via direct microscopic visualization, parasite isolation, and PCR. PCR reactions were positive in both clinical samples. Parasite isolation was possible for both patients, and isolates were submitted to species identification by isoenzyme electrophoresis and DNA sequencing. This report is the first to describe human infections caused by L. lindenbergi since the initial description and record of human infection by this species in 2002.

          Translated abstract

          La leishmaniose tégumentaire (TL) en Amazonie brésilienne est associée à plusieurs espèces de Leishmania. Dans ce rapport, nous décrivons deux cas de TL liés à Leishmania lindenbergi survenus à différents endroits de l’état de Rondônia. Après le diagnostic clinique, des échantillons de lésion ont été recueillis à des fins de diagnostic parasitologique par visualisation au microscope direct, isolement du parasite et PCR. Les réactions de PCR étaient positives dans les deux échantillons cliniques. L’isolement des parasites a été possible pour les deux patients et les isolats ont été soumis à l’identification de l’espèce par électrophorèse des isoenzymes et séquençage de l’ADN. Ce rapport est le premier à décrire les infections humaines causées par L. lindenbergi depuis la description initiale et le signalement de cas d’infection humaine par cette espèce en 2002.

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          Development and validation of PCR-based assays for diagnosis of American cutaneous leishmaniasis and identification of the parasite species.

          In this study, PCR assays targeting different Leishmania heat-shock protein 70 gene (hsp70) regions, producing fragments ranging in size from 230-390 bp were developed and evaluated to determine their potential as a tool for the specific molecular diagnosis of cutaneous leishmaniasis (CL). A total of 70 Leishmania strains were analysed, including seven reference strains (RS) and 63 previously typed strains. Analysis of the RS indicated a specific region of 234 bp in the hsp70 gene as a valid target that was highly sensitive for detection of Leishmania species DNA with capacity of distinguishing all analyzed species, after polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). This PCR assay was compared with other PCR targets used for the molecular diagnosis of leishmaniasis: hsp70 (1400-bp region), internal transcribed spacer (ITS)1 and glucose-6-phosphate dehydrogenase (G6pd). A good agreement among the methods was observed concerning the Leishmania species identification. Moreover, to evaluate the potential for molecular diagnosis, we compared the PCR targets hsp70-234 bp, ITS1, G6pd and mkDNA using a panel of 99 DNA samples from tissue fragments collected from patients with confirmed CL. Both PCR-hsp70-234 bp and PCR-ITS1 detected Leishmania DNA in more than 70% of the samples. However, using hsp70-234 bp PCR-RFLP, identification of all of the Leishmania species associated with CL in Brazil can be achieved employing a simpler and cheaper electrophoresis protocol.
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            Further Evidence of an Association between the Presence of Leishmania RNA Virus 1 and the Mucosal Manifestations in Tegumentary Leishmaniasis Patients

            Tegumentary Leishmaniasis (TL) is endemic in Latin America, and Brazil contributes approximately 20 thousand cases per year. The pathogenesis of TL, however, is still not fully understood. Clinical manifestations vary from cutaneous leishmaniasis (CL) to more severe outcomes, such as disseminated leishmaniasis (DL), mucosal leishmaniasis (ML) and diffuse cutaneous leishmaniasis (DCL). Many factors have been associated with the severity of the disease and the development of lesions. Recent studies have reported that the presence of Leishmania RNA virus 1 infecting Leishmania (Leishmania RNA virus 1, LRV1) is an important factor associated with the severity of ML in experimental animal models. In the present study, 156 patients who attended Rondonia's Hospital of Tropical Medicine with both leishmaniasis clinical diagnoses (109 CL; 38 ML; 5 CL+ML; 3 DL and 1 DCL) and molecular diagnoses were investigated. The clinical diagnosis were confirmed by PCR by targeting hsp70 and kDNA DNA sequences and the species causing the infection were determined by HSP70 PCR-RFPL. The presence of LVR1 was tested by RT-PCR. Five Leishmania species were detected: 121 (77.6%) samples were positive for Leishmania (Viannia) braziliensis, 18 (11.5%) were positive for Leishmania (V.) guyanensis, 3 (1.8%) for Leishmania (V.) lainsoni, 2 (1.3%) for Leishmania (Leishmania) amazonensis and 2 (1.3%) for Leishmania (V.) shawi. Six (3.9%) samples were positive for Leishmania sp. but the species could not be determined, and 4 (2.6%) samples were suggestive of mixed infection by L. (V.) braziliensis and L. (V.) guyanensis. The virus was detected in L. braziliensis (N = 54), L. guyanensis (N = 5), L. amazonensis (N = 2), L. lainsoni (N = 1) and inconclusive samples (N = 6). Patients presenting with CL+ML, DL and DCL were excluded from further analysis. Association between the presence of the virus and the disease outcome were tested among the remaining 147 patients (CL = 109 and ML = 38). Of them, 71.1% (n = 27) mucosal lesions were positive for LRV1, and 28.9% (n = 11) were negative. In cutaneous lesions, 36.7% (n = 40) were positive and 63.3% (n = 69) were negative for LRV1. The ratio P(ML|LRV1+)/P(ML|LRV1-) was 2.93 (CI95% 1.57…5.46; p<0.001), thus corroborating the hypothesis of the association between LRV1 and the occurrence of mucosal leishmaniasis, as previously described in animal models; it also indicates that LRV1 is not the only factor contributing to the disease outcome.
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              The Neotropical Leishmania species: a brief historical review of their discovery, ecology and taxonomy

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                Author and article information

                Journal
                Parasite
                Parasite
                parasite
                Parasite
                EDP Sciences
                1252-607X
                1776-1042
                2019
                23 May 2019
                : 26
                : ( publisher-idID: parasite/2019/01 )
                : 30
                Affiliations
                [1 ] Fundação Oswaldo Cruz, Unidade Rondônia Porto Velho Rondônia Brazil
                [2 ] Fundação Oswaldo Cruz, Instituto Oswaldo Cruz, IOC Rio de Janeiro Brazil
                [3 ] Secretaria de Saúde de Rondônia, SESAU Porto Velho Rondônia Brazil
                Author notes
                [* ]Corresponding author: elisa.cupolillo@ 123456gmail.com
                Author information
                http://orcid.org/0000-0002-2376-4236
                http://orcid.org/0000-0002-3514-3031
                http://orcid.org/0000-0003-2363-1289
                http://orcid.org/0000-0002-0620-3250
                Article
                parasite190012 10.1051/parasite/2019030
                10.1051/parasite/2019030
                6532396
                31120019
                b34f0e12-ae26-4d70-be31-1727b1d031aa
                © L.M. Cantanhêde et al., published by EDP Sciences, 2019

                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.

                History
                : 05 February 2019
                : 23 April 2019
                Page count
                Figures: 4, Tables: 1, Equations: 0, References: 14, Pages: 5
                Categories
                Short Note

                leishmania (viannia) lindenbergi,tegumentary leishmaniasis,western amazon

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