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Hist. ciênc. saúde-Manguinhos ; 27(supl.1): 95-122, Sept. 2020. tab
Article in English | LILACS | ID: biblio-1134089


Abstract The first autochthonous cases of cutaneous and mucocutaneous leishmaniasis in the Americas were described in 1909, but visceral leishmaniasis only erupted as a public health problem in the region in 1934. Today Brazil is the country with the most cases of American tegumentary leishmaniasis, and alongside India has the highest incidence of visceral leishmaniasis. Knowledge production and efforts to control these diseases have mobilized health professionals, government agencies and institutions, international agencies, and rural and urban populations. My research addresses the exchange and cooperation networks they established, and uncertainties and controversial aspects when notable changes were made in the approach to the New World leishmaniases.

Resumo Os primeiros casos de leishmaniose cutânea e mucocutânea autóctones das Américas foram descritos em 1909, e em 1934 a leishmaniose visceral irrompeu como problema de saúde pública na região. O Brasil tem hoje o maior número de casos da leishmaniose tegumentar americana e, junto com a Índia, a mais elevada incidência de leishmaniose visceral. A produção de conhecimentos e os esforços para controlar essas doenças mobilizaram, em nível global, profissionais de saúde, populações urbanas e rurais, instituições governamentais e agências internacionais. Recuperam-se aqui alguns desses agrupamentos, redes de troca e cooperação, incertezas e polêmicas, identificando-se mudanças na abordagem das leishmanioses do Novo Mundo.

Humans , History, 20th Century , Public Health/history , Leishmaniasis, Cutaneous/history , Leishmania , Tropical Medicine/history , Americas/epidemiology , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Cutaneous/prevention & control , Leishmaniasis, Cutaneous/epidemiology
An. bras. dermatol ; 95(4): 459-468, July-Aug. 2020. tab, graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS | ID: biblio-1130899


Abstract Background American cutaneous leishmaniasis is an infectious dermatosis caused by protozoa of the genus Leishmania, which comprises a broad spectrum of clinical manifestations depending on the parasite species involved in the infections and the immunogenetic response of the host. The use of techniques for amplification of the parasites DNA based on polymerase chain reaction polymerase chain reaction and the recent application of combined techniques, such as high-resolution DNA dissociation, have been described as a viable alternative for the detection and identification of Leishmania spp. in biological samples. Objectives To identify the Leishmania species using the polymerase chain reaction high-resolution DNA dissociation technique in skin biopsies of hospital-treated patients, and compare with results obtained by other molecular identification techniques. Methods A retrospective study assessing patients with suspected American cutaneous leishmaniasis seen at a hospital in São Paulo/Brazil was conducted. The paraffin blocks of 22 patients were analyzed by polymerase chain reaction high-resolution DNA dissociation to confirm the diagnosis and identify the species. Results Of the 22 patients with suspected American cutaneous leishmaniasis, the parasite was identified in 14, comprising five cases (35.6%) of infection by L. amazonensis, four (28.5%) by L. braziliensis, two (14.4%) by L. amazonensis + L. infantum chagasi, two (14.4%) by L. guyanensis, and one (7.1%) by Leishmania infantum chagasi. In one of the samples, in which the presence of amastigotes was confirmed on histopathological examination, the polymerase chain reaction high-resolution DNA dissociation technique failed to detect the DNA of the parasite. Study limitations The retrospective nature of the study and small number of patients. Conclusions The method detected and identified Leishmania species in paraffin-embedded skin biopsies with a sensitivity of 96.4% and could be routinely used in the public health system.

Humans , Leishmaniasis, Cutaneous/parasitology , Leishmania , United States , Brazil , Leishmaniasis, Mucocutaneous , Retrospective Studies , Leishmania infantum
Mem. Inst. Oswaldo Cruz ; 115: e190431, 2020. tab, graf
Article in English | SES-SP, LILACS, SES-SP | ID: biblio-1135241


BACKGROUND Long lasting insecticide-treated nets (LLINs) may be effective for vector control of cutaneous leishmaniasis (CL). Their efficacy, however, has not been sufficiently evaluated. OBJECTIVE To evaluate the large-scale efficacy of LLINs on Lutzomyia longiflocosa entomological parameters up to two years post-intervention in the sub-Andean region of Colombia. METHODS A matched-triplet cluster-randomised study of 21 rural settlements, matched by pre-intervention L. longiflocosa indoor density was used to compare three interventions: dip it yourself (DIY) lambda-cyhalothrin LLIN, deltamethrin LLIN, and untreated nets (control). Sand fly indoor density, feeding success, and parity were recorded using CDC light trap collections at 1, 6, 12, and 24 months post-intervention. FINDINGS Both LLINs reduced significantly (74-76%) the indoor density and the proportion of fully engorged sand flies up to two years post-intervention without differences between them. Residual lethal effects of both LLINs and the use of all nets remained high throughout the two-year evaluation period. CONCLUSIONS Both LLINs demonstrated high efficacy against L. longiflocosa indoors. Therefore, the deployment of these LLINs could have a significant impact on the reduction of CL transmission in the sub-Andean region. The DIY lambda-cyhalothrin kit may be used to convert untreated nets to LLINs increasing coverage.

Animals , Mosquito Control/methods , Leishmaniasis, Cutaneous/prevention & control , Insecticide-Treated Bednets , Insect Vectors/drug effects , Insecticides/administration & dosage , Anopheles/drug effects , Rural Population , Insecticide Resistance , Leishmaniasis, Cutaneous/parasitology , Colombia , Mosquito Vectors
Rev. Soc. Bras. Med. Trop ; 53: e20200091, 2020. graf
Article in English | ColecionaSUS, LILACS, ColecionaSUS, SES-SP | ID: biblio-1136875


Abstract INTRODUCTION: The drugs currently available for leishmaniasis treatment have major limitations. METHODS: In vitro and in vivo studies were performed to evaluate the effect of a quinoline derivative, Hydraqui (7-chloro-4-(3-hydroxy-benzilidenehydrazo)quinoline, against Leishmania amazonensis. In silico analyses of absorption, distribution, metabolism, excretion, and toxicity (ADMET) parameters were performed. RESULTS: Hydraqui showed significant in vitro anti-amastigote activity. Also, Hydraqui-treated mice exhibited high efficacy in lesion size (48.3%) and parasitic load (93.8%) reduction, did not cause hepatic and renal toxicity, and showed appropriate ADMET properties. CONCLUSIONS: Hydraqui presents a set of satisfactory criteria for its application as an antileishmanial agent.

Animals , Female , Quinolines/therapeutic use , Leishmania mexicana/drug effects , Leishmaniasis, Cutaneous/drug therapy , Antiprotozoal Agents/therapeutic use , Quinolines/chemistry , Leishmaniasis, Cutaneous/parasitology , Disease Models, Animal , Parasite Load , Mice , Mice, Inbred BALB C
Rev. Soc. Bras. Med. Trop ; 53: e20190139, 2020. graf
Article in English | LILACS | ID: biblio-1057288


Abstract INTRODUCTION: Leishmaniasis, a disease caused by a parasite endemic to large areas of tropical and subtropical countries, is a growing public health problem. METHODS: Male BALB/c mice were infected with Leishmania amazonensis and treated with extracts isolated from Annona mucosa. RESULTS: Treated groups had significantly reduced footpad swelling. The group treated intraperitoneally with hexane extract showed footpad swelling similar to groups treated with Pentamidine® and Glucantime®. Groups treated with dichloromethane extract and hexane extract presented the recovering phenotype associated with reduced parasite levels. CONCLUSIONS: Extracts of A. mucosa are promising sources of novel antileishmanial compounds.

Animals , Male , Plant Extracts/therapeutic use , Leishmaniasis, Cutaneous/drug therapy , Annona/chemistry , Leishmania/drug effects , Antiprotozoal Agents/therapeutic use , Leishmaniasis, Cutaneous/parasitology , Disease Models, Animal , Mice , Mice, Inbred BALB C , Antiprotozoal Agents/isolation & purification
Mem. Inst. Oswaldo Cruz ; 114: e190111, 2019. tab, graf
Article in English | LILACS | ID: biblio-1020081


BACKGROUND In addition to the limited therapeutic arsenal and the side effects of antileishmanial agents, drug resistance hinders disease control. In Brazil, Leishmania braziliensis causes atypical (AT) tegumentary leishmaniasis lesions, frequently refractory to treatment. OBJECTIVES The main goal of this study was to characterise antimony (Sb)-resistant (SbR) L. braziliensis strains obtained from patients living in Xakriabá indigenous community, Minas Gerais, Brazil. METHODS The aquaglyceroporin 1-encoding gene (AQP1) from L. braziliensis clinical isolates was sequenced, and its function was evaluated by hypo-osmotic shock. mRNA levels of genes associated with Sb resistance were measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Atomic absorption was used to measure Sb uptake. FINDINGS Although clinical isolates presented delayed recovery time in hypo-osmotic shock, AQP1 function was maintained. Isolate 340 accumulated less Sb than all other isolates, supporting the 65-fold downregulation of AQP1 mRNA levels. Both 330 and 340 isolates upregulated antimony resistance marker (ARM) 56/ARM58 and multidrug resistant protein A (MRPA); however, only ARM58 upregulation was an exclusive feature of SbR field isolates. CA7AE seemed to increase drug uptake in L. braziliensis and represented a tool to study the role of glycoconjugates in Sb transport. MAIN CONCLUSIONS There is a clear correlation between ARM56/58 upregulation and Sb resistance in AT-harbouring patients, suggesting the use of these markers as potential indicators to help the treatment choice and outcome, preventing therapeutic failure.

Humans , Leishmania braziliensis/drug effects , Leishmania braziliensis/genetics , Drug Resistance/drug effects , Leishmaniasis, Cutaneous/parasitology , Aquaglyceroporins/metabolism , Antimony/pharmacology , Drug Resistance/genetics , Real-Time Polymerase Chain Reaction
Braz. j. infect. dis ; 22(4): 278-287, July-Aug. 2018. tab, graf
Article in English | LILACS | ID: biblio-974226


ABSTRACT Background Leishmania major is a causative agent of zoonotic cutaneous leishmaniasis in the center of Iran, Abarkouh district. Molecular characterization and precise incrimination of Leishmania species was carried out to perform controlling measurements and to design treatment programs for zoonotic cutaneous leishmaniasis. Methods All smears isolated from ulcers of suspected patients were examined under a light microscope and graded for amastigotes frequency. Extraction of DNA, PCR, RFLP and sequencing of ITS-rDNA genotype were done to increase the efficacy of Leishmania parasites identification at their species-specific level and to detect any Leishmania infections within. Results Humans were found to be infected with L. major with high infection frequency and also Leishmania tropica was identified with low occurrence for the first time as non-native species using molecular analyses. The rates of infections was considerable with microscopic observation (n= 65, 73%) out of 89 smears prepared from suspected patients. Molecular analyses showed that the density of L. major was significantly higher (n= 48, 53.93%) than L. tropica (n= 4, 4.49%) (Mann-Whitney U test: p< 0.05) and two samples (2.25%) remained ambiguous after several sequencing. L. major did not have diversity with two common haplotypes but L. tropica were found to exhibit high diversity with three novel haplotypes. Conclusion L. major was considered the causative agent of leishmaniasis in the region, but the identification of a non-native L. tropica revealed the importance of further isolation of Leishmania parasites following molecular analyses and confirmation, and also revealed the importance of further isolation of Leishmania parasites from patients of the field areas who do not have easily access to health care centers for specialized treatment strategies.

Humans , Animals , Male , Female , Leishmania tropica/genetics , Leishmaniasis, Cutaneous/parasitology , Leishmania major/genetics , Rural Population , Haplotypes , Polymorphism, Restriction Fragment Length , Leishmania tropica/isolation & purification , Leishmania tropica/ultrastructure , Polymerase Chain Reaction , DNA, Protozoan/isolation & purification , DNA, Protozoan/genetics , Leishmaniasis, Cutaneous/pathology , Leishmaniasis, Cutaneous/epidemiology , Leishmania major/isolation & purification , Endemic Diseases , Iran
Mem. Inst. Oswaldo Cruz ; 113(4): e170250, 2018. tab
Article in English | LILACS | ID: biblio-1040594


Several studies have described the use of non-invasive collection methods, mostly based on the detection of parasite DNA, for diagnosis. However, no Leishmania specimens have been isolated from saliva. Here, we report the first isolation of Leishmania braziliensis from the saliva of humans with cutaneous leishmaniasis but without lesions on their mucosa. The isolates were obtained from salivary fluid inoculated in hamsters and were tested by multilocus enzyme electrophoresis. Seven samples from 43 patients suspected of having the disease were identified for in vivo culture. These findings suggest that saliva is a clinical sample that allows the isolation of Leishmania sp.

Humans , Male , Female , Adolescent , Adult , Young Adult , Saliva/parasitology , Leishmania braziliensis/isolation & purification , Disease Reservoirs , Polymerase Chain Reaction , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/parasitology , Endemic Diseases , Electrophoresis , Middle Aged
Mem. Inst. Oswaldo Cruz ; 112(7): 517-519, July 2017.
Article in English | LILACS | ID: biblio-841810


The sandfly Trichophoromyia auraensis has recently evolved as a proven vector of Leishmania (Viannia) endemic to state of Acre in the north of Brazil. This note is intended to propose a correction in the report of the first occurrence of natural infection of Leishmania (Viannia) in this species. We and the other scientific groups reinforced that Tr. auraensis is a possible vector involved in the transmission of American cutaneous leishmaniasis in Acre, Brazil.

Animals , Leishmaniasis, Cutaneous/parasitology , Insect Vectors/classification , Insect Vectors/parasitology , Leishmania/isolation & purification , Psychodidae , Psychodidae/parasitology , Brazil
Mem. Inst. Oswaldo Cruz ; 112(1): 44-52, Jan. 2017. tab, graf
Article in English | LILACS | ID: biblio-841754


Leishmania are protozoan parasites that show remarkable diversity, as revealed by the various clinical forms of leishmaniasis, which can range from mild skin lesions to severe metastatic cutaneous/mucosal lesions. The exact nature and extent of Leishmania phenotypic diversity in establishing infection is not fully understood. In order to try to understand some aspects of this diversity, we subcutaneously infected BALB/c mice with first and second generation subclones of a L. amazonensis strain isolated from a patient (BA125) and examined in vivo lesion growth rate and antimony susceptibility. In vivo fast-, medium- and slow-growing subclones were obtained; moreover, fast-growing subclones could generate slow-growing subclones and inversely, revealing the continuous generation of diversity after passage into mice. No antimony-resistant subclone appeared, probably a rare occurrence. By tagging subclone cells with a L. amazonensis genomic cosmid library, we found that only a very small number of founding cells could produce lesions. Leishmania clones transfected with in vivo selected individual cosmids were also diverse in terms of lesion growth rate, revealing the cosmid-independent intrinsic characteristics of each clone. Our results suggest that only a few of the infecting parasites are able to grow and produce lesions; later, within the cell mixture of each lesion, there coexist several parasite populations with different potentialities to grow lesions during the next infection round. This may reflect a sort of programmed heterogeneity of individual parasites, favoring the survival of some individuals in various environmental conditions.

Animals , Female , Leishmania mexicana/genetics , Leishmania mexicana/pathogenicity , Leishmaniasis, Cutaneous/parasitology , Disease Models, Animal , Phenotype , Time Factors , Mice, Inbred BALB C
Mem. Inst. Oswaldo Cruz ; 111(7): 460-468, tab, graf
Article in English | LILACS | ID: lil-787557


The 70 kDa heat shock protein (HSP70) is a molecular chaperone that assists the parasite Leishmania in returning to homeostasis after being subjected to different types of stress during its life cycle. In the present study, we evaluated the effects of HSP70 transfection of L. amazonensis promastigotes (pTEX-HSP70) in terms of morphology, resistance, infectivity and mitochondrial bioenergetics. The pTEX-HSP70 promastigotes showed no ultrastructural morphological changes compared to control parasites. Interestingly, the pTEX-HSP70 promastigotes are resistant to heat shock, H2O2-induced oxidative stress and hyperbaric environments. Regarding the bioenergetics parameters, the pTEX-HSP70 parasites had higher respiratory rates and released less H2O2 than the control parasites. Nevertheless, the infectivity capacity of the parasites did not change, as verified by the infection of murine peritoneal macrophages and human macrophages, as well as the infection of BALB/c mice. Together, these results indicate that the overexpression of HSP70 protects L. amazonensis from stress, but does not interfere with its infective capacity.

Animals , Female , HSP70 Heat-Shock Proteins/physiology , Leishmania mexicana/physiology , Leishmaniasis, Cutaneous/parasitology , Protozoan Proteins/physiology , Stress, Physiological , HSP70 Heat-Shock Proteins/genetics , Leishmania mexicana/genetics , Leishmania mexicana/ultrastructure , Macrophages/parasitology , Mice , Mice, Inbred BALB C , Mitochondria/physiology , Oxidative Stress , Protozoan Proteins/genetics , Transfection/methods
An. bras. dermatol ; 91(3): 365-367, graf
Article in English | LILACS | ID: lil-787294


Abstract: We report an imported case of cutaneous leishmaniasis in a 37-year-old man from Saudi Arabia caused by Leishmania major. He presented with non-healing nodulo-ulcerative lesions with a "volcanic crater" on the lower limbs. It was clearly cutaneous leishmaniasis - a rare disease in China - as reflected by the patient's clinical history, the lesions' morphology, histopathological examination, culture and PCR analysis of the lesions. The patient was completely cured after two cycles of sodium stibogluconate treatment. This case report demonstrates that dermatologists should be aware of sporadic cutaneous leishmaniasis cases in non-endemic areas.

Humans , Male , Adult , Leishmaniasis, Cutaneous/parasitology , Leishmania major , Saudi Arabia , China/ethnology , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/drug therapy , Antimony Sodium Gluconate/therapeutic use , Emigrants and Immigrants , Leg Ulcer/parasitology , Antiprotozoal Agents/therapeutic use
Rev. bras. parasitol. vet ; 25(1): 127-130, Jan.-Mar. 2016. tab, graf
Article in English | LILACS | ID: lil-777540


Abstract The skin is the site of inoculation of Leishmania spp. in susceptible hosts, and consequently dermatopathies, especially ulcerative dermatitis, are the main clinical signs observed. The aim of this study was to assess parasitism of the skin (intact and ulcerated) among dogs that were naturally infected by Leishmania spp., through immunohistochemical analysis. Skin fragments (intact and ulcerated) were collected from 13 dogs with positive parasitological (bone marrow aspiration and exfoliative skin) and serological examinations (ELISA S7® Biogene) forLeishmania spp. These samples were processed using the immunohistochemical technique, involving the streptavidin-peroxidase complex. Ulcerative lesions were mainly observed on the elbows (53.84%; 7/13), nostrils (15.38%; 2/13), ears (23.07%; 3/13) and wings of the ilium (7.69%; 1/13). A severe parasite load was detected in 46.15% and 76.92% of the intact and ulcerated skin samples tested, respectively. The parasite load on ulcerated skin was statistically higher than on intact skin (p = 0.0221). These results indicate that the intact and ulcerated skin may host a high parasite load of amastigote forms of Leishmania spp., which can favor the transmission of the parasite.

Resumo A pele é o local de inoculação da Leishmania spp. nos hospedeiros susceptíveis e dessa forma, as dermatopatias, principalmente as dermatites ulcerativas são os principais sinais clínicos observados. O objetivo deste estudo foi avaliar o parasitismo na pele (íntegra e ulcerada) em cães naturalmente infectados por Leishmania spp. através da técnica de imunohistoquímica. Fragmentos de pele (íntegra e ulcerada) foram coletados de 13 cães com diagnóstico parasitológico (aspirado de medula óssea e esfoliação cutânea) e sorológico positivos (ELISA S7® Biogene) paraLeishmania spp. Amostras foram processadas por imunohistoquímica pelo complexo estreptoavidina-peroxidase. As lesões ulcerativas foram observadas principalmente nas regiões do cotovelo 53,84% (7/13), narina 15,38% (2/13), orelha 23,07% (3/13) e sobre a asa do ílio 7,69% (1/13). Uma intensa carga parasitária foi detectada 46,15% e 76,92% das amostras de pele íntegra e ulcerada, respectivamente. A carga parasitária na pele ulcerada foi estatisticamente superior à pele íntegra (p = 0,0221). Esses resultados indicam que a pele intacta e ulcerada pode albergar uma intensa carga parasitária de formas amastigotas de Leishmania spp., o que pode favorecer a transmissão do parasita.

Animals , Dogs , Skin/parasitology , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Cutaneous/veterinary , Parasite Load/veterinary , Dog Diseases/parasitology
Salvador; s.n; 2016. 88 p. ilus, tab.
Thesis in Portuguese | LILACS | ID: biblio-1001024


As leishmanioses constituem um complexo de doenças causada pelo protozoário intracelular, do gênero Leishmania, sendo a resposta imune celular essencial para controle, eliminação e proteção contra a infecção. A teoria clonal da imunidade celular propõe que as respostas imunológicas são estabelecidas através do aumento na frequência de clones específicos ao antígeno. Para avaliar a resposta das células T à infecção por Leishmania, investigamos, por citometria de fluxo, a expressão de cadeias Vβ de receptores de células T (TCRs), estado de ativação, capacidade de adesão ao endotélio e potencial funcional de clones específico. Em um grupo de pacientes com Leishmaniose Cutânea Localizada (LCL), avaliamos diferentes subpopulações de células T através da expressão da região Vβ, no sangue periférico e na biópsia da lesão. Utilizamos células mononucleares de sangue periférico (CMSPs), de pacientes LCL e controles saudáveis, nas quais avaliamos, ex vivo, a expressão de moléculas de ativação (CD25, CD69 e HLA-DR), adesão (LFA-1, VLA-4 e CD62L), co-estimulatória (CD27 e CD28)...

Leishmaniasis is a desease caused by infection with the Leishmania protozoan parasite. The cellular immune response is essential for controlling, eliminating and protection of the Leishmania infection. The clonal theory of cellular immunity proposes that immunological responses are established by increasing the frequency of antigen-specific clones. In order to measure the host T cell response to Leishmania infection, we have investigated by flow cytometry, the expression of Vβ chains of T-cell receptors (TCRs), activation state, adhesion to endothelium of capacity and functional potential of specific T. In a group of localized cutaneous leishmaniasis (LCL) patients, we evaluated different T cell subpopulations as identified by their Vβ region expression, in peripheral blood and biopsy. We used peripheral blood mononuclear cells (PBMCs), from CL patients and healthy volunteers, in which we evaluate, ex vivo, the expression of activation molecules (CD25, CD69 and HLA-DR), adhesion (LFA-1, VLA-4 and CD62L), co-stimulatory (CD27 and CD28)...

Humans , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/epidemiology , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Cutaneous/pathology , Leishmaniasis, Cutaneous/prevention & control , Lymphocytes/immunology , Lymphocytes/microbiology , Lymphocytes/pathology
Mem. Inst. Oswaldo Cruz ; 110(8): 1017-1023, Dec. 2015. tab, graf
Article in English | LILACS | ID: lil-769836


Leishmania donovani is the known causative agent of both cutaneous (CL) and visceral leishmaniasis in Sri Lanka. CL is considered to be under-reported partly due to relatively poor sensitivity and specificity of microscopic diagnosis. We compared robustness of three previously described polymerase chain reaction (PCR) based methods to detectLeishmania DNA in 38 punch biopsy samples from patients presented with suspected lesions in 2010. Both, Leishmaniagenus-specific JW11/JW12 KDNA and LITSR/L5.8S internal transcribed spacer (ITS)1 PCR assays detected 92% (35/38) of the samples whereas a KDNA assay specific forL. donovani (LdF/LdR) detected only 71% (27/38) of samples. All positive samples showed a L. donovani banding pattern upon HaeIII ITS1 PCR-restriction fragment length polymorphism analysis. PCR assay specificity was evaluated in samples containing Mycobacterium tuberculosis, Mycobacterium leprae, and human DNA, and there was no cross-amplification in JW11/JW12 and LITSR/L5.8S PCR assays. The LdF/LdR PCR assay did not amplify M. leprae or human DNA although 500 bp and 700 bp bands were observed in M. tuberculosis samples. In conclusion, it was successfully shown in this study that it is possible to diagnose Sri Lankan CL with high accuracy, to genus and species identification, using Leishmania DNA PCR assays.

Humans , DNA, Protozoan/isolation & purification , Leishmania donovani/genetics , Leishmaniasis, Cutaneous/parasitology , Polymerase Chain Reaction/methods , Skin/parasitology , Biopsy , DNA Primers , Leishmaniasis, Cutaneous/pathology , Neglected Diseases/parasitology , Polymorphism, Restriction Fragment Length , Polymerase Chain Reaction/standards , Sensitivity and Specificity , Species Specificity , Sri Lanka , Skin/pathology
Rev. Inst. Med. Trop. Säo Paulo ; 57(5): 451-454, Sept.-Oct. 2015. graf
Article in English | LILACS | ID: lil-766276


SUMMARY American tegumentary leishmaniasis (ATL) is an infectious disease caused by protozoa of the genus Leishmania, and transmitted by sandflies. In the state of Rio de Janeiro, almost all of the cases of American tegumentary leishmaniasis (ATL) are caused by Leishmania (Viannia) braziliensis, while cases of visceral leishmaniasis (VL) are caused by Leishmania (Leishmania) infantum chagasi. The resurgence of autochthonous VL cases in Rio de Janeiro is related to the geographic expansion of the vector Lutzomyia longipalpis and its ability to adapt to urban areas. We report the first case of leishmaniasis with exclusively cutaneous manifestations caused by L. (L.) infantum chagasi in an urban area of Rio de Janeiro. An eighty-one-year-old woman presented three pleomorphic skin lesions that were not associated with systemic symptoms or visceromegalies. Multilocus enzyme electrophoresis identified L. (L.) infantum chagasi, but direct smear and PCR of bone narrow were negative for Leishmania sp. (suggesting exclusively cutaneous involvement). We discuss the different dermatological presentations of viscerotropic leishmaniasis of the New and Old World, and the clinical and epidemiological importance of the case. Etiologic diagnosis of ATL based upon exclusive clinical criteria may lead to incorrect conclusions. We should be aware of the constant changes in epidemiological patterns related to leishmaniases.

RESUMO A leishmaniose tegumentar americana (LTA) é uma doença infecciosa causada por protozoários do gênero Leishmania, transmitida por flebotomíneos. No estado do Rio de Janeiro, quase todos os casos de LTA são causados por Leishmania (Viannia) braziliensis, enquanto a leishmaniose visceral (LV) é causada por Leishmania (Leishmania) infantum chagasi. O ressurgimento de casos autóctones de LV no Rio de Janeiro está relacionado com a expansão geográfica dos vetores Lutzomyia longipalpis e à sua capacidade de se adaptar às áreas urbanas. Relatamos o primeiro caso de leishmaniose com manifestações exclusivamente cutâneas causadas por L. (L.) infantum chagasiem uma área urbana do Rio de Janeiro. Mulher de 81 anos apresentou três lesões cutâneas pleomórficas não associadas a sintomas sistêmicos ou visceromegalias. A eletroforese de enzimas multilocus obtida a partir da lesão cutânea identificou L. (L.) infantum chagasi,por outro lado o exame direto e o PCR da medula óssea foram negativos para Leishmaniasp. (sugerindo acometimento exclusivamente cutâneo). Discutimos as diferentes apresentações dermatológicos da leishmaniose visceral do Novo e Velho Mundo, assim como a importância clínica e epidemiológica deste caso. O diagnóstico etiológico da LTA com base apenas em critérios clínicos pode levar a conclusões incorretas. Devemos estar conscientes das constantes mudanças nos padrões epidemiológicos relacionados à leishmaniose.

Aged, 80 and over , Animals , Female , Humans , Leishmania infantum/genetics , Leishmaniasis, Cutaneous/parasitology , Brazil/epidemiology , Electrophoresis, Polyacrylamide Gel , Leishmania infantum/isolation & purification , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/epidemiology , Polymerase Chain Reaction
Rev. Soc. Bras. Med. Trop ; 48(5): 560-567, Sept.-Oct. 2015. graf
Article in English | LILACS | ID: lil-763329


ABSTRACTINTRODUCTION:The aim of this study was quantify annexin A1 expression in macrophages and cluster of differentiation 4 (CD4) + and cluster of differentiation 8 (CD8)+ T cells from the skin of patients with cutaneous leishmaniasis (n=55) and correlate with histopathological aspects.METHODS:Infecting species were identified by polymerase chain reaction-restriction fragment length polymorphism, and expression of annexin A1 was analyzed by immunofluorescence.RESULTS:All patients (n = 55) were infected with Leishmania braziliensis . Annexin A1 was expressed more abundantly in CD163 + macrophages in infected skin (p < 0.0001) than in uninfected skin. In addition, macrophages in necrotic exudative reaction lesions expressed annexin A1 at higher levels than those observed in granulomatous (p < 0.01) and cellular lesions p < 0.05). This difference might be due to the need to clear both parasites and necrotic tissue from necrotic lesions. CD4 + cells in cellular lesions expressed annexin A1 more abundantly than did those in necrotic (p < 0.05) and granulomatous lesions (p < 0.01). Expression in CD8 + T cells followed the same trend. These differences might be due to the pervasiveness of lymphohistiocytic and plasmacytic infiltrate in cellular lesions.CONCLUSIONS:Annexin A1 is differentially expressed in CD163 + macrophages and T cells depending on the histopathological features of Leishmania -infected skin, which might affect cell activation.

Female , Humans , Male , Middle Aged , Annexin A1/metabolism , Leishmania/classification , Leishmaniasis, Cutaneous/metabolism , Leishmaniasis, Cutaneous/pathology , Annexin A1/analysis , Cross-Sectional Studies , Fluorescent Antibody Technique , Leishmaniasis, Cutaneous/parasitology , Macrophages/metabolism , Macrophages/parasitology , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length
Rev. Inst. Med. Trop. Säo Paulo ; 57(4): 343-347, July-Aug. 2015. tab, ilus
Article in English | LILACS | ID: lil-761162


SUMMARYIn this study, Leishmaniaspecies were identified by Polymerase Chain Reaction (PCR). The epidemiology of patients suspected of having American Cutaneous Leishmaniasis in the municipality of Assis Brasil, Acre State, located in the Brazil/Peru/Bolivia triborder was also investigated. By PCR, the DNA of Leishmaniawas detected in 100% of the cases (37 samples) and a PCR-Restriction Fragment Length Polymorphism (RFLP) of the hsp 70gene identified the species in 32 samples: Leishmania (Viannia) braziliensis (65.6%) , L. (V.) shawi (28.1%) , L. (V.) guyanensis (3.1%) and mixed infection L. (V.) guyanensisand L. (Leishmania) amazonensis(3.1%)This is the first report of L. (V.) shawiand L. (L.) amazonensisin Acre. The two predominant species were found in patients living in urban and rural areas. Most cases were found in males living in rural areas for at least three years and involved in rural work. This suggests, in most cases, a possible transmission of the disease from a rural/forest source, although some patients had not engaged in activities associated with permanence in forestall areas, which indicate a possible sandflies adaptation to the periurban setting.

RESUMOO presente estudo caracterizou as espécies de Leishmaniapela Reação em Cadeia da Polimerase (PCR). Também descreveu os aspectos epidemiológicos de pacientes com suspeita de leishmaniose tegumentar americana do município de Assis Brasil, Estado do Acre, Brasil, localizado na tríplice fronteira Brasil/Peru/Bolívia. A PCR detectou DNA de Leishmaniaem 100% dos casos (37 amostras) e a PCR- Restriction Fragment Length Polymorfism(RFLP) do gene hsp 70identificou as espécies em 32 amostras: Leishmania (Viannia) braziliensis (65,6%) , L. (V.) shawi (28,1%) , L. (V.) guyanensis (3,1%) e infecção mista L. (V.) guyanensise L. (Leishmania) amazonensis(3,1%)Esse é o primeiro registro de L. (V.) shawie L. (L.) amazonensisno Acre. As duas espécies predominantes foram encontradas em indivíduos residentes em áreas rurais e urbanas. O maior número de casos foi notificado entre indivíduos de áreas rurais, sexo masculino, de ocupação rural e tempo de residência maior que três anos. Esses dados sugerem possível transmissão da doença em ambiente rural/florestal na maioria dos casos, no entanto alguns pacientes não tinham envolvimento com atividades relacionadas com a permanência na floresta, indicando possível adaptação de flebotomíneos no ambiente periurbano.

Humans , Animals , Male , Female , Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Adult , Young Adult , Leishmania/classification , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Mucocutaneous/parasitology , Brazil/epidemiology , Leishmania/genetics , Leishmania/isolation & purification , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/epidemiology , Leishmaniasis, Mucocutaneous/diagnosis , Leishmaniasis, Mucocutaneous/epidemiology , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Risk Factors , Rural Population , Urban Population
Braz. j. infect. dis ; 19(3): 302-307, May-Jun/2015. tab, graf
Article in English | LILACS | ID: lil-751876


Background: Several tests are performed to obtain better accuracy when diagnosing American tegumentary leishmaniasis (ATL). It is believed that antigens released via secretion, excretion and metabolism are more specific than are antigens released by the lysis of Leishmania parasites. Such antigens are known as exo-antigens (exo-Ag) and are formed from products released by cultured parasites in a way that is similar to that in which they cause infections in hosts. Objective: We attempted to validate a Leishmania mexicana ELISA exo-Ag for ATL diagnosis in Midwestern Brazil. Methods: A total of 281 patients were included in the study. We analysed pre-treatment blood from 98 ATL patients; out of those, 85.7% and 14.3% had cutaneous and mucosal forms, respectively. Results: The exo-Ag accuracy was 83.99% (95% CI = 79.24-87.81) with a sensitivity value of 90.82% (95% CI = 83.46-95.09) and an overall specificity value of 80.33% (95% CI = 73.97-85.44). The positive predictive value and negative predictive value were 71.20% (95% CI = 62.72-78.41) and 94.23% (95% CI = 89.40-96.94), respectively. Among healthy controls, exo-Ag had a specificity of 91.25% (95% CI = 83.02-95.70); additionally, the test had specificity rates of 66.67% (95% CI = 46.71-82.03) in Chagas disease patients, 60.61% (95% CI = 43.68-75.32) in patients with rheumatic diseases, 76.92% (95% CI = 49.74-91.82) in pemphigus foliaceus patients, 87.50% (95% CI = 52.91-97.76) in leprosy patients, 87.50% (95% CI = 63.98-96.50) in VRDL-positive patients, and 77.78 (95% CI = 45.26-93.68) in deep mycosis patients. Conclusion: Based on the indicators of validity, we conclude that the results obtained in this study enable the recommendation of the exo-Ag ELISA for ATL diagnosis once it presented a reasonable accuracy compared to classical methods. Cost evaluations are necessary to completely define the role of this technique in large scale. .

Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Antigens, Protozoan/blood , Enzyme-Linked Immunosorbent Assay/methods , Leishmania braziliensis/immunology , Leishmania mexicana/immunology , Leishmaniasis, Cutaneous/diagnosis , Case-Control Studies , Leishmaniasis, Cutaneous/parasitology , Predictive Value of Tests , Sensitivity and Specificity
Rev. Inst. Med. Trop. Säo Paulo ; 57(3): 257-262, May-Jun/2015. tab, graf
Article in English | LILACS | ID: lil-752598


Introduction: American tegumentary leishmaniasis (ATL) can be caused by Leishmania (Viannia) braziliensis complex. The evolution of ATL initially results in lesions and can develop into disseminated or diffuse forms. The genetic diversity of L. (V.) braziliensis in some endemic areas of Brazil has been poorly studied, such as in the state of São Paulo. This study analyzed the genetic diversity of L. (V.) braziliensis isolates collected from patients and dogs with LTA from the state of São Paulo. Methods: Leishmaniasis diagnosis was determined by PCR. The 132 biopsies were collected in different regions of Sao Paulo State, Brazil (36 municipalities). The genetic characterization of L. (V.) braziliensis isolates was tested by RFLP-PCR using DNA extracted from biopsies. The primer set amplified a specific region of Leishmania internal transcribed spacers of the ribosomal DNA locus. Results: Of the 132 samples, 52 (40%) were completely genotyped by RFLP-PCR (44 from human patients and eight from dogs). The results showed nine distinct patterns. The majority of the genotyped samples were from Sorocaba (30), and the others were distributed among 14 other municipalities. The first pattern was more frequent (29 samples), followed by pattern 2 (nine samples) and pattern 3 (three samples). Patterns 4, 6, 7, 8 and 9 were composed of two samples each and pattern 5 of one sample. Conclusion: These results suggest that polymorphic strains of L. (V.) braziliensis circulate in the state of São Paulo. These data agree with studies from other regions of Brazil, showing great variability among the natural populations of endemic foci. .

Introdução: A leishmaniose tegumentar americana (LTA) é causada pelo sub-gênero Leishmania (Viannia) braziliensis. A evolução da LTA resulta com a evolução das lesões iniciais. A diversidade genética de L. (V.) braziliensis em algumas áreas endêmicas brasileiras, como no estado de São Paulo, é pouco conhecida. Assim, este estudo teve como objetivo analisar a variabilidade genética de isolados de L. (V.) braziliensis coletados de biopsias de pacientes e cães com LTA no estado de São Paulo. Métodos: O diagnóstico da leishmaniose foi realizado por PCR. As 132 biópsias analisadas foram coletadas em diferentes regiões do Estado de São Paulo, Brasil (36 municípios). A caracterização genética de L. (V.) braziliensis foi realizada por RFLP-PCR utilizando DNA extraído das biopsias. O conjunto de iniciadores utilizado amplificou a região ITS de Leishmania. Resultados: Das 132 amostras analisadas, 52 (40%) foram completamente genotipadas por RFLP-PCR (44 de pacientes e oito de cães). Os resultados mostraram nove padrões distintos. A maioria das amostras genotipadas foi de Sorocaba (30), e as demais foram distribuídas entre 14 outros municípios. O primeiro padrão foi mais frequente (29 amostras), seguido pelo padrão 2 (nove amostras), padrão 3 (três amostras). Padrões 4, 6, 7, 8 e 9 foram compostos de duas amostras de cada um e o padrão 5, com uma amostra. Conclusão: Estes resultados sugerem que cepas polimórficas de L. (V.) braziliensis circulam no estado de São Paulo. Estes dados são concordantes com estudos em outras regiões do Brasil, mostrando grande variabilidade destas populações naturais de focos endêmicos. .

Animals , Dogs , Humans , DNA, Protozoan/genetics , Genetic Variation , Leishmania braziliensis/genetics , Leishmaniasis, Cutaneous/parasitology , Biopsy , Brazil , Genotype , Leishmania braziliensis/isolation & purification , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length