RESUMEN
Cutaneous leishmaniasis (CL) stands out as a significant vector-borne endemic in Pakistan. Despite the rising incidence of CL, the genetic diversity of Leishmania species in the country's endemic regions remains insufficiently explored. This study aims to uncover the genetic diversity and molecular characteristics of Leishmania species in CL-endemic areas of Baluchistan, Khyber Pakhtunkhwa (KPK), and Punjab in Pakistan. Clinical samples from 300 CL patients were put to microscopic examination, real-time ITS-1 PCR, and sequencing. Predominantly affecting males between 16 to 30 years of age, with lesions primarily on hands and faces, the majority presented with nodular and plaque types. Microscopic analysis revealed a positivity rate of 67.8%, while real-time PCR identified 60.98% positive cases, mainly L. tropica, followed by L. infantum and L. major. Leishmania major (p = 0.009) showed substantially greater variation in nucleotide sequences than L. tropica (p = 0.07) and L. infantum (p = 0.03). Nucleotide diversity analysis indicated higher diversity in L. major and L. infantum compared to L. tropica. This study enhances our understanding of CL epidemiology in Pakistan, stressing the crucial role of molecular techniques in accurate species identification. The foundational data provided here emphasizes the necessity for future research to investigate deeper into genetic diversity and its implications for CL control at both individual and community levels.
Asunto(s)
Variación Genética , Leishmaniasis Cutánea , Leishmaniasis Cutánea/epidemiología , Leishmaniasis Cutánea/parasitología , Pakistán/epidemiología , Humanos , Masculino , Adolescente , Adulto , Femenino , Adulto Joven , Niño , Persona de Mediana Edad , Leishmania/genética , Leishmania/clasificación , Leishmania/aislamiento & purificación , Preescolar , Análisis de Secuencia de ADN , Leishmania tropica/genética , Leishmania tropica/aislamiento & purificación , Leishmania tropica/clasificación , Leishmania major/genética , Leishmania major/clasificación , Leishmania major/aislamiento & purificación , ADN Protozoario/genética , Filogenia , Epidemiología Molecular , Anciano , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
Cutaneous leishmaniasis is atypical in Sri Lanka because Leishmania donovani, which typically causes visceral disease, is the causative agent. The origins of recently described hybrids between L. donovani and other Leishmania spp. usually responsible for cutaneous leishmaniasis remain unknown. Other endemic dermotropic Leishmania spp. have not been reported in Sri Lanka. Genome analysis of 27 clinical isolates from Sri Lanka and 32 Old World Leishmania spp. strains found 8 patient isolates clustered with L. tropica and 19 with L. donovani. The L. tropica isolates from Sri Lanka shared markers with strain LtK26 reported decades ago in India, indicating they were not products of recent interspecies hybridization. Because L. tropica was isolated from patients with leishmaniasis in Sri Lanka, our findings indicate L. donovani is not the only cause of cutaneous leishmaniasis in Sri Lanka and potentially explains a haplotype that led to interspecies dermotropic L. donovani hybrids.
Asunto(s)
Leishmania tropica , Leishmaniasis Cutánea , Secuenciación Completa del Genoma , Sri Lanka , Humanos , Leishmania tropica/genética , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/parasitología , Leishmaniasis Cutánea/diagnóstico , Filogenia , Genoma de Protozoos , Masculino , Femenino , Leishmania donovani/genética , Leishmania donovani/aislamiento & purificación , Adulto , Persona de Mediana EdadRESUMEN
According to the World Health Organization (WHO), leishmaniasis is a zoonotic/anthroponotic parasitic disease endemic in 99 countries. It is estimated that approximately 12 million people are infected with Leishmania spp. and 350 million people live at risk. Every year, two million new cases are added to these figures. One and a half million cases of zoonotic/anthroponotic cutaneous leishmaniasis and 500 000 cases of visceral leishmaniasis are reported annually. One person is estimated to to be infected with cutaneous leishmaniasis in every 20 seconds and visceral leishmaniasis causes 60 000 deaths. In this report, two pediatric cases diagnosed with visceral leishmaniasis were presented. In the study, bone marrow aspirations were performed to determine the etiology of the disease in an eight-month-old male patient with fever and hepatosplenomegaly who had been followed up in Manisa Celal Bayar University, Department of Pediatrics, Division of Pediatric Hematology with the diagnosis of severe glucose-6-phosphate dehydrogenase (G-6PD) deficiency since the neonatal period and in a nine-month-old female patient who had had a high fever and bicytopenia for two weeks. Bone marrow aspirations were cultured in NNN medium and their smears were stained and examined with Giemsa. rk-39 and Leishmania IFAT tests were performed by using patients' sera. Quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) analysis was also performed for Leishmania spp. Leishmania spp. amastigotes were observed in Giemsa-stained smear preparations, Leishmania spp. promastigotes were grown in NNN medium, rk39 rapid diagnostic kit was weakly positive, Leishmania IFAT was positive at a titer of 1/1024 and Leishmania tropica was identified as the causative agent by RT-qPCR analysis for both cases. These two cases suggested that fatal cases of visceral leishmaniasis may increase with the spread of visceralized isolates of L.tropica, the most common causative agent of cutaneous leishmaniasis in Türkiye, and this issue may create a significant public health problem.
Asunto(s)
Leishmania tropica , Leishmaniasis Visceral , Humanos , Masculino , Leishmaniasis Visceral/diagnóstico , Leishmaniasis Visceral/parasitología , Leishmania tropica/aislamiento & purificación , Leishmania tropica/genética , Femenino , Lactante , Médula Ósea/parasitología , Deficiencia de Glucosafosfato Deshidrogenasa/complicaciones , AnimalesRESUMEN
Cutaneous leishmaniasis (CL), a neglected tropical disease, is a major public health concern in Yemen, with Leishmania tropica identified as the main causative agent. This study aims to investigate the occurrence and distribution of Leishmania parasites in domestic and wild animals in CL endemic areas in the western highlands of Yemen. A cross-sectional study was conducted in the Utmah District of western Yemen. Blood and skin scraping specimens were collected from 122 domestic and wild animals and tested for the Leishmania DNA using internal transcribed spacer 1 (ITS1) nested polymerase chain reaction. Phylogenetic analyses were performed on 20 L. tropica sequences obtained from animals in this study and 34 sequences from human isolates (collected concurrently from the same study area) retrieved from the GenBank. Overall, L. tropica was detected in 16.4% (20/122) of the examined animals, including 11 goats, two dogs, two bulls, one cow, one donkey, one rabbit, one rat and one bat. None of the examined cats and sheep was positive. The animal sequences were segregated into four different L. tropica haplotypes, with the majority of the animal (15/20) and human (32/34) sequences composed of one dominant haplotype/genotype. These findings represent the first confirmed evidence of natural L. tropica infections in different kinds of domestic and wild animals in western Yemen, suggesting these animals potentially have a role in the transmission of CL in Yemen. Therefore, a One Health approach is required for the effective prevention and control of this devastating disease among endemic populations.
Asunto(s)
Animales Domésticos , Animales Salvajes , Leishmania tropica , Leishmaniasis Cutánea , Salud Única , Filogenia , Animales , Leishmania tropica/genética , Leishmania tropica/aislamiento & purificación , Leishmania tropica/clasificación , Leishmaniasis Cutánea/epidemiología , Leishmaniasis Cutánea/veterinaria , Leishmaniasis Cutánea/parasitología , Yemen/epidemiología , Humanos , Estudios Transversales , Animales Salvajes/parasitología , Animales Domésticos/parasitología , ADN Protozoario/genética , Enfermedades Desatendidas/parasitología , Enfermedades Desatendidas/epidemiología , Enfermedades Desatendidas/veterinaria , Enfermedades Endémicas/veterinaria , ADN Espaciador Ribosómico/genética , Reacción en Cadena de la Polimerasa/veterinaria , Análisis de Secuencia de ADN , MasculinoRESUMEN
This study investigated nine provinces in northern Morocco and collected 275 skin scraping, 22 bone marrow aspirates, and 89 fine needle aspirations from suspected cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL) patients and potentially infected dogs. Molecular analysis using ITS1 RFLP PCR and RT-PCR revealed a higher prevalence of L. infantum (66.18 %; χ2 = 28.804; df = 1; P-value = 8.01e-08) than L. tropica in skin scraping, with L. infantum being the sole causative agent for both VL and canine leishmaniasis. L. infantum was predominantly found in most provinces, while L. tropica was relatively more dominant in Taza Province. Discriminant Analysis of Principal Components (DAPC) revealed distinct clustering between L. tropica and the other three species. However, no small subset of SNPs could clearly differentiate between Infantum_CL, Infantum_VL, and CanL, as they likely share a significant genetic background. The high rate of L. infantum could be attributed to the abundance of sand fly species transmitting VL. In Taza Province, Phlebotomus sergenti, responsible for anthroponotic CL, is the most abundant species. DNA sequencing demonstrated sequence heterogeneity in L. infantum (variants 1-9) and L. tropica (variants 1-7). Phylogenetic analysis showed a distinct separation between L. tropica and L. infantum strains, with an overlap among L. infantum strains isolated from cutaneous, visceral, and canine cases, and dogs serving as the central population for L. infantum.
Asunto(s)
Enfermedades de los Perros , Variación Genética , Leishmania infantum , Leishmania tropica , Leishmaniasis Visceral , Perros , Animales , Leishmania infantum/genética , Leishmania infantum/aislamiento & purificación , Leishmania tropica/genética , Leishmania tropica/aislamiento & purificación , Marruecos , Humanos , Enfermedades de los Perros/parasitología , Enfermedades de los Perros/genética , Leishmaniasis Visceral/veterinaria , Leishmaniasis Visceral/parasitología , Leishmaniasis Visceral/epidemiología , Leishmaniasis Cutánea/parasitología , Leishmaniasis Cutánea/veterinaria , Leishmaniasis Cutánea/epidemiología , Filogenia , Masculino , Polimorfismo de Nucleótido SimpleRESUMEN
PURPOSE: Iran is among the high-risk leishmaniasis regions in the world. WHO recommends the use of GIS as an ideal tool for healthcare authorities to predict the evolution of a disease, delimit the risk of outbreaks and identify critical areas. The aim of this research is to find the association between the main species of Leishmania (L. major, L. tropica, L. infantum) dispersion and climatic variables in Iran. METHODS: All molecular-based reports of leishmaniasis from Iran between 1999 and 2021 were gathered from reliable medical sources. Meteorological data (air and soil temperatures, annual rainfall and humidity) of the country along the study period were obtained from the Iranian Climatological Research Centre. The data concerning species distribution and climatic conditions during this period were moved to a base-map through raster layers using ArcGIS 10.4.1 software. The relationship between parasitological and climatic models was examined using ANOVA. RESULTS: High risk area maps, based on the cut-off thresholds, were generated for Leishmania major, L. tropica and L. infantum. According to the molecular-based reports, the L. major distribution was significantly related to all climatic variables, while L. tropica was merely related to rainfall and humidity, and the L. infantum distribution was significantly associated with rainfall, soil and air temperatures. CONCLUSION: The association between climatic conditions and Leishmania species distribution in Iran has been confirmed. Consequently, both, the relationship between climatic conditions and the geographical distribution of Leishmania species, and the use of GIS to better understand the spatial epidemiology of leishmaniasis, have been reaffirmed.
Asunto(s)
Clima , Irán/epidemiología , Humanos , Leishmaniasis/epidemiología , Leishmania/clasificación , Leishmania/aislamiento & purificación , Leishmania infantum/aislamiento & purificación , Sistemas de Información Geográfica , Temperatura , Leishmania tropica/aislamiento & purificación , Leishmania major/aislamiento & purificaciónRESUMEN
BACKGROUND: The kinetoplastid protozoan Leishmania tropica mainly causes cutaneous leishmaniasis in humans in the Middle East, and relapse or treatment failure after treatment are common in this area. L. tropica's digenic life cycle includes distinct stages in the vector sandfly and the mammalian host. Sexual reproduction and genetic exchange appear to occur more frequently than in other Leishmania species. Understanding these processes is complicated by chromosome instability during cell division that yields aneuploidy, recombination and heterozygosity. This combination of rare recombination and aneuploid permits may reveal signs of hypothetical parasexual mating, where diploid cells fuse to form a transient tetraploid that undergoes chromosomal recombination and gradual chromosomal loss. METHODOLOGY/PRINCIPAL FINDINGS: The genome-wide SNP diversity from 22 L. tropica isolates showed chromosome-specific runs of patchy heterozygosity and extensive chromosome copy number variation. All these isolates were collected during 2007-2017 in Sweden from patients infected in the Middle East and included isolates from a patient possessing two genetically distinct leishmaniasis infections three years apart with no evidence of re-infection. We found differing ancestries on the same chromosome (chr36) across multiple samples: matching the reference genome with few derived alleles, followed by blocks of heterozygous SNPs, and then by clusters of homozygous SNPs with specific recombination breakpoints at an inferred origin of replication. Other chromosomes had similar marked changes in heterozygosity at strand-switch regions separating polycistronic transcriptional units. CONCLUSION/SIGNIFICANCE: These large-scale intra- and inter-chromosomal changes in diversity driven by recombination and aneuploidy suggest multiple mechanisms of cell reproduction and diversification in L. tropica, including mitotic, meiotic and parasexual processes. It underpins the need for more genomic surveillance of Leishmania, to detect emerging hybrids that could spread more widely and to better understand the association between genetic variation and treatment outcome. Furthering our understanding of Leishmania genome evolution and ancestry will aid better diagnostics and treatment for cutaneous leishmaniasis caused by L.tropica in the Middle East.
Asunto(s)
Genoma de Protozoos , Leishmania tropica/genética , Leishmaniasis Cutánea/parasitología , Afganistán , Cromosomas/genética , Variaciones en el Número de Copia de ADN , ADN Protozoario/genética , Variación Genética , Humanos , Irán , Leishmania tropica/clasificación , Leishmania tropica/aislamiento & purificación , Filogenia , Polimorfismo de Nucleótido Simple , Recombinación Genética , SiriaRESUMEN
Leishmaniasis is a neglected zoonotic disease that poses significant veterinary and public health risks in developing countries. Dogs act as a reservoir host for leishmaniasis transmitted to humans. A total of 108 human cases of cutaneous leishmaniasis (CL) were identified in the Al-Houd Al-Marsoud Hospital in Cairo, Egypt, during 2018. Blood samples and skin biopsies were collected for further examination. Blood samples from 96 asymptomatic dogs were collected. All samples were subjected to molecular and phylogenetic analysis. Quantitative RT-PCR was used to measure the expression of genes related to mTOR signalling and inflammation in blood and tissue samples. The distribution pattern of human cases pointed to an endemic focus in North Sinai (66.67%). The prevalence of asymptomatic canine leishmaniasis was 66.60%. Histopathological examination of human skin lesions revealed a severe granulomatous inflammatory reaction, necrosis and ulceration. Moreover, leishmanial amastigotes could be detected in human tissue samples. Phylogenetic analysis revealed 100% identity of human isolates to Leishmania tropica (MN453682), and dog isolates to Leishmania infantum (MN453673), with 94.9% similarity between the two isolates. Gene expression related to mTOR signalling and inflammation in both species' samples confirmed a significant alteration of EIF4EBP1, CCR4 and INF-γ expression compared with control groups. In Egypt, increased incidence of asymptomatic carrier dogs acting as a significant reservoir host for Leishmania poses a public health hazard. Findings warrant further epidemiological investigation of CL in Egypt, as well as additional study of parasite differentiation and gene regulation.
Asunto(s)
Enfermedades de los Perros/parasitología , Leishmania tropica , Leishmaniasis Cutánea/veterinaria , Leishmaniasis Visceral/veterinaria , Zoonosis/parasitología , Adulto , Animales , Reservorios de Enfermedades/parasitología , Reservorios de Enfermedades/veterinaria , Perros , Egipto/epidemiología , Femenino , Regulación de la Expresión Génica , Humanos , Leishmania infantum/genética , Leishmania infantum/aislamiento & purificación , Leishmania tropica/genética , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/epidemiología , Leishmaniasis Cutánea/parasitología , Leishmaniasis Visceral/epidemiología , Leishmaniasis Visceral/parasitología , Masculino , Persona de Mediana Edad , Proteínas Protozoarias/genética , Proteínas Protozoarias/metabolismo , Adulto Joven , Zoonosis/epidemiologíaRESUMEN
BACKGROUND: Epidemics of cutaneous leishmaniasis (CL) are occurring more frequently and spreading faster and farther than before in many areas of the world. The present study aimed to assess a long-lasting emerging epidemic (2005-2019) of 5532 cases with anthroponotic CL (ACL) in peri-urban areas of Kerman city in southeastern Iran. METHODS: This descriptive-analytical study was carried out for 15 years in Kerman province, southeastern Iran. The data were passively obtained through the health surveillance system and the Kerman Leishmaniasis Research Center. Every subject was diagnosed using direct smear microscopy. The representative causative agent was further examined by ITS1-PCR, PCR-RFLP, 7SL RNA gene sequencing and phylogenetic analyses. For each subject, a case report form designating demographic and clinical data was recorded. RESULTS: A different pattern of ACL incidence was found in peri-urban areas compared to that in the city of Kerman. The incidence rate of ACL cases has significantly increased (P < 0.001) from 2005 to 2016 in new settlements with a gradual decline after that. The overall average risk of contracting the disease was 7.6 times higher in peri-urban areas compared to Kerman city, an old endemic focus. All isolates consisting of six variants were confirmed to be Leishmania tropica. The overall pattern of the ACL infection indicates that the etiological agent of ACL is propagated and transmitted by the bite of female Phlebotomus sergenti sandflies from person to person from dissimilar clones as reflected by the complexity of the migrants' backgrounds in the province. CONCLUSIONS: The movement of populations and establishment of new settlements in peri-urban areas close to endemic areas are major risk factors for and are directly linked to CL. The underlying factors of this emerging ACL epidemic caused by L. tropica were disasters and droughts, among others. A robust commitment to a multilateral approach is crucial to make improvements in this area. This will require decisive coordinated actions through all governmental factions and non-governmental organizations. Furthermore, active and passive case detection strategies, early diagnosis, and effective treatment could help control the disease.
Asunto(s)
Leishmaniasis Cutánea/epidemiología , Leishmaniasis Cutánea/transmisión , Phlebotomus/fisiología , Adolescente , Adulto , Animales , Niño , Monitoreo Epidemiológico , Femenino , Humanos , Irán/epidemiología , Leishmania tropica/clasificación , Leishmania tropica/genética , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/parasitología , Masculino , Phlebotomus/parasitología , Filogenia , Factores de Riesgo , Migrantes/estadística & datos numéricos , Viaje/estadística & datos numéricos , Población Urbana/estadística & datos numéricos , Adulto JovenRESUMEN
Cutaneous leishmaniasis (CL) is a major health problem in Iran, with a heavy burden on human health and society. There is little knowledge about the molecular epidemiology of the disease, as well as phylogenetic relationship of causative agents in south-eastern Iran. The aim of the present study was to investigate the molecular aspects of CL, especially atypical CL in the Bam district, Kerman province, south-eastern Iran, as an endemic region of CL in Iran. The smears were collected from lesion samples of 353 patients clinically suspected to CL, who attended local health centres in the Bam district during 2016-2017. Direct smears were examined for Leishmania parasites using the Giemsa staining technique. Amplification of kinetoplast DNA (kDNA) and the ribosomal internal transcribed spacer 1(ITS-1) gene were carried out using polymerase chain reaction (PCR). Then, the ITS1-PCR products were sequenced for phylogenetic analysis. Overall, 278 cases were confirmed as CL by microscopic examination of Giemsa-stained slides. Clinical presentation of the lesions was basically of two types: (a) typical lesions and (b) atypical including lupoid ulcers, sporotrichoid, nodular and exudative lesions. The PCR assay on all specimens of skin lesions proved L. tropica as the main pathogenic agent. Phylogenic analysis revealed high similarity among isolates from the Bam district in the south-east with isolates from Birjand in eastern Iran, as well as with isolates from Herat province in western Afghanistan. The study provided valuable information concerning the genetic diversity of the parasite as one of the factors influencing the clinical manifestations in CL in south-eastern Iran, which could be the basis for planning future control strategies.
Asunto(s)
Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/parasitología , Adolescente , Adulto , Anciano , Niño , Preescolar , ADN de Cinetoplasto/análisis , ADN Espaciador Ribosómico/análisis , Femenino , Humanos , Lactante , Irán , Leishmania tropica/clasificación , Leishmania tropica/genética , Masculino , Persona de Mediana Edad , Filogenia , Adulto JovenRESUMEN
BACKGROUND: Drug resistance is a common phenomenon frequently observed in countries where leishmaniasis is endemic. Due to the production of the pteridine reductase enzyme (PTR1), drugs lose their efficacy, and consequently, the patient becomes unresponsive to treatment. This study aimed to compare the in vitro effect of meglumine antimoniate (MA) on non- healing Leishmania tropica isolates and on MA transfected non-healing one to PTR1. METHODS: Two non-healing and one healing isolates of L. tropica were collected from patients who received two courses or one cycle of intralesional MA along with biweekly liquid nitrogen cryotherapy or systemic treatment alone, respectively. After confirmation of L. tropica isolates by polymerase chain reaction (PCR), the recombinant plasmid pcDNA-rPTR (antisense) was transfected via electroporation and cultured on M199. Isolates in form of promastigotes were treated with different concentrations of MA and read using an enzyme-linked immunosorbent assay (ELISA) reader and the half inhibitory concentration (IC50 ) value was calculated. The amastigotes were grown in mouse macrophages and were similarly treated with various concentrations of MA. The culture glass slides were stained, and the mean number of intramacrophage amastigotes and infected macrophages were assessed in triplicate for both stages. RESULTS: All three transfected isolates displayed a reduction in optical density compared with the promastigotes in respective isolates, although there was no significant difference between non-healing and healing isolates. In contrast, in the clinical form (amastigotes), there was a significant difference between non-healing and healing isolates (p < 0.05). CONCLUSION: The results indicated that the PTR1 gene reduced the efficacy of the drug, and its inhibition by antisense and could improve the treatment of non-healing cases. These findings have future implications in the prophylactic and therapeutic modality of non- healing Leishmania isolates to drug.
Asunto(s)
Leishmania tropica/genética , Leishmaniasis Cutánea/tratamiento farmacológico , Leishmaniasis Cutánea/genética , Oxidorreductasas/genética , Proteínas Protozoarias/genética , Adulto , Animales , Antiprotozoarios/farmacología , Antiprotozoarios/uso terapéutico , ADN sin Sentido , Farmacorresistencia Microbiana/efectos de los fármacos , Farmacorresistencia Microbiana/genética , Femenino , Humanos , Leishmania tropica/efectos de los fármacos , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/parasitología , Masculino , Antimoniato de Meglumina/farmacología , Antimoniato de Meglumina/uso terapéutico , Ratones Endogámicos BALB C , TransfecciónRESUMEN
Since leishmaniases are zoonotic vector-borne diseases transmitted through the bites of infected female sand flies, identification of the sources of imbibed blood meals and the detection and identification of leishmanial DNA in them are important in discerning animal reservoirs, clarifying the epidemiology and facilitating control of local leishmaniases. CDC light traps, aspirators and sticky paper traps were used to collect sand flies in four Palestinian foci of both, CL and VL. Phlebotomine species identification was based on morphological keys. Female specimens were screened to detect and identify leishmanial infections, using kDNA-PCR and ITS1-PCR, and engorged female specimens were analyzed to identify the origin of their blood meals, using an RDB blood meal assay based on the amplification of the cytochrome b gene (cytb) of vertebrate mitochondrial DNA (mtDNA). Twenty sand fly species, 11 of the genus Phlebotomus and nine the genus Sergentomyia, were identified. The most abundant species was Ph. papatasi (33.7%), followed by Ph. sergenti (21%). Among the 691 female sand fly specimens, 18.5% (128/691) were positive for leishmanial DNA, using the kDNA-PCR and 6.4% (44/691) were positive using the ITS1-PCR. DNA from parasites of the genus Leishmania was identified in only 1.5% of the infected sand flies. That of Leishmania tropica parasites was detected in six female specimens of Ph. sergenti and that of L. major parasites in two female specimens of Ph. papatasi. Interestingly, two engorged females of the species Se. (Neophlebotomus) sp. were positive for L. tropica DNA. Ninety engorged female sand flies of Ph. papatasi and 104 of Ph. sergenti had fed on a large variety of vertebrate hosts such as humans, hyraxes, rats, cows, goats and birds. Regarding blood-meals showing a mixture from different species of animal host, hyrax and rat blood was revealed in 8/104 (7.7%) females of Ph. sergenti. Detection of hyrax blood in engorged female sand flies of the species Ph. sergenti supports the role of hyraxes being a potential reservoir of L. tropica in Palestinian regions. Rat blood meals might be significant since a few strains L. tropica and L. infantum were isolated from rats. Further studies must be undertaken before conclusions could be drawn.
Asunto(s)
ADN de Cinetoplasto/análisis , Conducta Alimentaria , Leishmania tropica/aislamiento & purificación , Psychodidae/fisiología , Animales , Árabes , ADN de Cinetoplasto/genética , ADN Espaciador Ribosómico/genética , Transmisión de Enfermedad Infecciosa , Femenino , Especificidad del Huésped , Humanos , Insectos Vectores/parasitología , Leishmania tropica/genética , Leishmaniasis/transmisión , Reacción en Cadena de la Polimerasa , Psychodidae/parasitologíaRESUMEN
Anthroponotic cutaneous leishmaniasis (CL) caused by Leishmania tropica (L. tropica) represents a public health challenge in several resource poor settings. We herein employed a systems analysis approach to study molecular signatures of CL caused by L. tropica in the skin lesions of ulcerative CL (UCL) and non-ulcerative CL (NUCL) patients. Results from RNA-seq analysis determined shared and unique functional transcriptional pathways in the lesions of the UCL and NUCL patients. Several transcriptional pathways involved in inflammatory response were positively enriched in the CL lesions. A multiplexed inflammatory protein analysis showed differential profiles of inflammatory cytokines and chemokines in the UCL and NUCL lesions. Transcriptional pathways for Fcγ receptor dependent phagocytosis were among shared enriched pathways. Using L. tropica specific antibody (Ab)-mediated phagocytosis assays, we could substantiate Ab-dependent cellular phagocytosis (ADCP) and Ab-dependent neutrophil phagocytosis (ADNP) activities in the lesions of the UCL and NUCL patients, which correlated with L. tropica specific IgG Abs. Interestingly, a negative correlation was observed between parasite load and L. tropica specific IgG/ADCP/ADNP in the skin lesions of CL patients. These results enhance our understanding of human skin response to CL caused by L. tropica.
Asunto(s)
Biomarcadores/análisis , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/diagnóstico , Carga de Parásitos/estadística & datos numéricos , RNA-Seq/métodos , Piel/patología , Estudios de Casos y Controles , Citocinas/análisis , Humanos , Leishmaniasis Cutánea/genética , Leishmaniasis Cutánea/inmunología , Leishmaniasis Cutánea/parasitología , Piel/metabolismo , Piel/parasitologíaRESUMEN
Antimony is an important drug for the treatment of Leishmania parasite infections. In several countries, the emergence of drug-resistant Leishmania species has reduced the effectiveness of this drug. The mechanism of clinical drug resistance is unclear. The aim of this work was to identify mitochondrial proteome alterations associated with resistance against antimonial. A combination of cell fractionation, liquid chromatography-tandem mass spectrometry (LC-MS/MS), and Label-Free Quantification was used to characterize the mitochondrial protein composition of Leishmania tropica field isolates resistant and sensitive to meglumine antimoniate. LC-MS/MS analysis resulted in the identification of about 1200 proteins of the Leishmania tropica mitochondrial proteome. Various criteria were used to allocate about 40% proteins to mitochondrial proteome. Comparative quantitative proteomic analysis of the sensitive and the resistant strains showed proteins with differential abundance in resistance species are involved in TCA and aerobic respiration enzymes, stress proteins, lipid metabolism enzymes, and translation. These results showed that the mechanism of antimony resistance in Leishmania spp. field isolate may be associated with alteration in enzymes involved in mitochondrial pathways.
Asunto(s)
Antiprotozoarios/farmacología , Leishmania tropica/efectos de los fármacos , Antimoniato de Meglumina/farmacología , Mitocondrias/metabolismo , Proteínas Mitocondriales/metabolismo , Animales , Línea Celular , Cromatografía Liquida , Resistencia a Medicamentos , Leishmania tropica/aislamiento & purificación , Leishmania tropica/metabolismo , Ratones , Mitocondrias/efectos de los fármacos , Pruebas de Sensibilidad Parasitaria , Proteoma , Proteómica , Espectrometría de Masas en TándemRESUMEN
Toscana virus (TOSV) is a prominent arthropod-borne viral agent of human central nervous system infections occurring in the Mediterranean region. The main transmission route to susceptible individuals involves sandflies as vectors. Despite several reports revealing widespread TOSV activity in Turkey, vectors remained unidentified. A sandfly field survey was carried out in five provinces in Central, Southeast and Mediterranean Anatolia in 2017 to identify TOSV and related sandfly-borne phleboviruses and Leishmania parasites, with evidence for circulation in the region. A total of 7136 sandfly specimens, collected via standard methods, were evaluated in 163 pools. TOSV was detected in 11 pools (6.7%), comprising Phlebotomus major sensu lato, Sergentomyia dentata and Phlebotomus papatasi species. TOSV partial L and S segment sequences were characterized, that phylogenetically clustered with local and global genotype A strains. An amino acid substitution outside the conserved motifs of the viral polymerase, also present in previous TOSV sequences in endemic regions, was observed. Leishmania tropica was detected in a single pool of Ph. sergentii (0.6%). This is the first report of TOSV in sandflies from Turkey, and this study further provides evidence for additional sandfly species with the potential to transmit TOSV.
Asunto(s)
Phlebotomus , Virus de Nápoles de la Fiebre de la Mosca de los Arenales , Animales , Infecciones por Bunyaviridae/transmisión , Humanos , Insectos Vectores/parasitología , Insectos Vectores/virología , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/transmisión , Phlebotomus/clasificación , Phlebotomus/parasitología , Phlebotomus/virología , Filogenia , Psychodidae/clasificación , Psychodidae/parasitología , Psychodidae/virología , ARN Viral , Virus de Nápoles de la Fiebre de la Mosca de los Arenales/genética , Virus de Nápoles de la Fiebre de la Mosca de los Arenales/aislamiento & purificación , Turquía/epidemiología , Enfermedades Transmitidas por Vectores/parasitología , Enfermedades Transmitidas por Vectores/transmisión , Enfermedades Transmitidas por Vectores/virologíaRESUMEN
Leishmaniasis is a parasitic disease infecting animals and humans. Two clinical forms (Visceral and cutaneous leishmaniasis) and four species are reported to be present in Turkey. Several studies have investigated canine and human leishmaniasis in Turkey but no study was performed to screen the infection among wild rodents, so far. The present study aims to investigate the role of small wild rodents as reservoir animals for Leishmania spp. in different regions of Turkey. Formalin-preserved tissue samples (spleen, liver, lung) of 712 rodents from 30 provinces were screened for the presence of Leishmania spp. DNA. Before DNA extraction, tissues were dried, rehydrated, and homogenated. Leishmania screening in rodent tissues and species determination was performed with a combination of real-time kDNA and ITS1 polymerase chain reaction protocols. Eight (1.12%) out of 712 animals were found to be positive for Leishmania spp. DNA and species typing revealed five L. infantum, two L. tropica and one L. major among positives. Leishmania major and L. infantum DNA were detected in Apodemus spp. from Zonguldak province located in the Western Black Sea Region, while L. tropica DNA was found in Meriones sp. and Gerbillus dasyurus from Adana and Hatay provinces located in Eastern Mediterranean Region of Turkey. The present study is first to report natural infection of L. infantum, L. major and L. tropica in small wild rodents in Turkey, suggesting their possible roles as reservoirs. Further studies are needed for planning epidemiological studies and also for developing rodent control measures in risky endemic areas to break the transmission cycle.
Asunto(s)
Reservorios de Enfermedades/veterinaria , Gerbillinae , Leishmaniasis Cutánea/veterinaria , Leishmaniasis Visceral/veterinaria , Murinae , Enfermedades de los Roedores/epidemiología , Animales , Femenino , Leishmania infantum/aislamiento & purificación , Leishmania major/aislamiento & purificación , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/epidemiología , Leishmaniasis Cutánea/parasitología , Leishmaniasis Visceral/epidemiología , Leishmaniasis Visceral/parasitología , Masculino , Prevalencia , Enfermedades de los Roedores/parasitología , Turquía/epidemiologíaRESUMEN
The province of Khorasan-Razavi in the North East of Iran is an endemic area for anthroponotic cutaneous leishmaniasis (ACL caused mainly by Leishmania tropica) and zoonotic cutaneous leishmaniasis (ZCL caused mainly by Leishmania major). Based on clinical signs, some cities were considered as ACL foci while others were considered to be endemic for ZCL. This paper reviews studies performed on patients diagnosed with cutaneous leishmaniasis (CL) via the use of direct slide examination, ELISA, electrophoresis isoenzyme, RAPD PCR and PCR in Mashhad; the study also includes cases of CL in other cities of the Khorasan-Razavi province where only PCR used as a diagnostic tool. The data show that both Leishmania tropica and Leishmania major caused CL in most of the cities investigated. Our review shows that Leishmania major was found in areas where ACL is prevalent and Leishmania tropica was observed in areas with high incidence of ZCL. This distribution represents a major change in the epidemiological pattern of Leishmania in the Khorasan-Razavi province.
Asunto(s)
Leishmania major/aislamiento & purificación , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/epidemiología , Zoonosis/parasitología , Adulto , Animales , Electroforesis/métodos , Ensayo de Inmunoadsorción Enzimática , Humanos , Incidencia , Irán/epidemiología , Leishmania major/genética , Leishmania tropica/genética , Leishmaniasis Cutánea/diagnóstico , Leishmaniasis Cutánea/parasitología , Reacción en Cadena de la Polimerasa , Prevalencia , Técnica del ADN Polimorfo Amplificado Aleatorio , Zoonosis/epidemiologíaRESUMEN
Objective: Leishmania tropica (L. tropica) and Leishmania infantum (L. infantum) are the species causing cutaneous Leishmaniasis (CL) in Turkey. There was a wave of immigration due civil war in Syria in 2011. Migration from Syria, where CL is endemic, to other countries is thought to affect the number of CL cases and species diversity. The aim of the study was to typify the samples of CL positive, pre-migration and post-migration Turkish patients and importe (Syrian) patients whose smears were found in the archive and to reveal the difference of CL species before and after migration in Hatay. Methods: Smears of a total of 150 patients (50 Turkish patients before migration, 50 Turkish patients after migration and 50 Syrian patients) which had been prepared with dermal scraping, stained with Giemsa and determined as CL positive by microscope examination were included in the study. DNA isolation of selected preparations was performed and GZ-PZR analysis with ITS-1probe was performed for species determination. Results: L. infantum/donovani was detected in 40 (80%), L. tropica in 8 (16%), and L. major in 2 (4%) of the samples belonging to pre-immigration Turkish patients. L. infantum/donovani was detected in 28 (56%), L. major in 3 (6%) and L. tropica in 19 (%38) of the samples belonging to post-immigration Turkish patients. L. infantum/donovani was detected in 2 (4%), L. major in 1 (2%) and L. tropica in 47 (94%) of the samples belonging to Syrian patients. Conclusions: It was observed that in local cases in Hatay before immigration, L. infantum/donovani was the common species that caused CL and that after immigration L. tropica began to raise and that L. major was more encountered than before. It was concluded that Syrians coming to Hatay may have caused diversity in the Leishmania species which were the causative agents of CL, and that further research was needed on the subject.
Asunto(s)
Técnicas de Genotipaje , Leishmania infantum/aislamiento & purificación , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/parasitología , Conflictos Armados , Colorantes Azulados , Enfermedades Endémicas , Femenino , Genotipo , Humanos , Leishmania infantum/clasificación , Leishmania infantum/genética , Leishmania major/clasificación , Leishmania major/genética , Leishmania major/aislamiento & purificación , Leishmania tropica/clasificación , Leishmania tropica/genética , Leishmaniasis Cutánea/diagnóstico , Leishmaniasis Cutánea/epidemiología , Masculino , Reacción en Cadena en Tiempo Real de la Polimerasa , Siria/epidemiología , Siria/etnología , Migrantes , Turquía/epidemiologíaAsunto(s)
Enfermedades de la Conjuntiva/parasitología , Infecciones Parasitarias del Ojo/parasitología , Leishmania tropica/genética , Leishmaniasis Cutánea/parasitología , Adulto , Anfotericina B/uso terapéutico , Animales , Antiprotozoarios/uso terapéutico , Enfermedades de la Conjuntiva/tratamiento farmacológico , ADN Protozoario/genética , Infecciones Parasitarias del Ojo/tratamiento farmacológico , Humanos , Leishmania tropica/aislamiento & purificación , Leishmaniasis Cutánea/tratamiento farmacológico , MasculinoRESUMEN
BACKGROUND & OBJECTIVES: Cutaneous leishmaniasis (CL) in Marrakesh-Safi region located in the central-south part of Morocco is a public health problem. This study assessed the efficiency of a microscopic examination method in establishing the diagnosis of CL and PCR for the characterization and identification of the circulating Leishmania strains in different CL foci of the study area. METHODS: A total of 297 smears obtained from cutaneous lesions of suspected patients with CL were stained with May-Grünwald Giemsa (MGG) for microscopic examination. For each positive smear, genomic DNA was extracted and PCR-analysed, targeting the small subunit ribosomal ribonucleic acid (ssu rRNA) gene to detect Leishmania DNA. Then, the internal transcribed spacer 1 (ITS1) was amplified and sequenced in order to identify the Leishmania species. The sensitivity and specificity of the conventional microscopy with ssu rRNA gene were compared by Leishmania nested PCR (LnPCR) and ITS1 gene by ITS-PCR. RESULTS: A total of 257 smears were positive in the microscopic examination, i.e. the detection rate of amastigotes by optical microscopy was 86.53% (257/297). The LnPCR was found to have a specificity and a sensitivity of 100%, each. Interestingly, the sequencing results showed that 99.61% (256/257) of the isolates had Leishmania tropica and 0.39% (1/257) had L. infantum infection. INTERPRETATION & CONCLUSION: Though, classical microscopic examination is useful and economical, it is not sensitive enough, especially in endemic regions where several Leishmania species coexist. In such situations, PCR constitutes a complementary method for the identification of the causal species. The results indicate that both the L. tropica (dominant) and L. infantum are the causative agents of CL in the Marrakesh-Safi region. The rate of CL infection is high in Imintanout, and Chichaoua provinces. Hence, early diagnosis and prompt treatment of CL patients is necessary to prevent its extension to neighboring localities.