ABSTRACT
BACKGROUND: Dogs are considered the main domestic animals that may be a reservoir for Leishmania infantum, the agent of zoonotic visceral leishmaniasis (ZVL) in several countries of the world. The dog may host other Leishmania species, but its epidemiological role in the maintenance and spreading of these parasites is not completely elucidated. Zoonotic cutaneous leishmaniasis (ZCL), caused by Leishmania major, affects thousands of people every year and is particularly diffused in many countries of North Africa and Middle East Asia. In ZCL endemic countries, few reports of L. major-positive dogs have been reported, probably because most human cases occur in poor rural areas where the social role of the dog and its medical management is not well considered. The aim of the present study is to better understand the possible involvement of domestic dogs in the epidemiology of ZCL. METHODS: Our research focused on a well-established endemic focus of ZCL, in the area of Echrarda, Kairouan Governorate, central Tunisia. A total of 51 dogs with no or mild clinical signs of vector borne diseases were selected in small villages where human cases of ZCL are yearly present. All dogs were sampled for the Leishmania spp. diagnosis, by using the following procedures: blood sample for serology and buffy coat quantitative polymerase chain reaction (qPCR), popliteal fine needle aspiration, and cutaneous biopsy punch for lymph node and skin qPCR. RESULTS: The results demonstrated a high percentage (21.6%) of dogs positive at least at one or more test; the most sensitive technique was the lymph node qPCR that detected 8/11 positive dogs. Nine, out of the eleven positive dogs, resulted as infected by Leishmania infantum; ITS1-PCR-sequencing allowed Leishmania major identification in the remaining two cases, both from the popliteal lymph node samples, which can suggest a possible visceral spread of a cutaneous Leishmania species in the dog. Interestingly, one of the two L. major-positive dogs was living in the same house where 6-year-old children showed cutaneous lesions referred to as ZCL. CONCLUSIONS: To our knowledge, this is the first report of L. major-positive dogs in Tunisia, the epidemiological role of which remains under investigation.
Subject(s)
Dog Diseases , Leishmania major , Leishmaniasis, Cutaneous , Zoonoses , Dogs , Animals , Leishmania major/isolation & purification , Leishmania major/genetics , Tunisia/epidemiology , Leishmaniasis, Cutaneous/epidemiology , Leishmaniasis, Cutaneous/veterinary , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Cutaneous/diagnosis , Dog Diseases/epidemiology , Dog Diseases/parasitology , Zoonoses/epidemiology , Zoonoses/parasitology , Zoonoses/transmission , Humans , Endemic Diseases/veterinary , Female , Male , Disease Reservoirs/parasitology , Disease Reservoirs/veterinaryABSTRACT
Cutaneous leishmaniasis (CL) is often considered a 'great imitator' and is the most common form of leishmaniasis. The Leishmania species responsible for CL varies among countries, as these species exhibit specific distribution patterns. The increased mobility of people across countries has resulted in the imported incidences of leishmaniasis caused by non-endemic species of Leishmania. During 2023, we confirmed three CL cases caused by L. major from Kerala, India, and upon detailed investigation, these were identified to be imported from the Middle East and Kazakhstan regions. This is the first report of CL caused by L. major from Kerala. The lesion morphology, detection of anti-rK 39 antibody and Leishmania parasite DNA from the blood samples were the unique observations of these cases. Kerala, being an emerging endemic zone of visceral leishmaniasis (VL) and CL, the imported incidences of leishmaniasis by non-endemic species can pose a significant threat, potentially initiating new transmission cycles of leishmaniasis caused by non-endemic species.
Subject(s)
Leishmania major , Leishmaniasis, Cutaneous , India/epidemiology , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Cutaneous/epidemiology , Leishmaniasis, Cutaneous/diagnosis , Humans , Male , Leishmania major/isolation & purification , Leishmania major/genetics , Adult , Female , Communicable Diseases, Imported/parasitology , Communicable Diseases, Imported/epidemiology , Middle Aged , DNA, Protozoan/genetics , Antibodies, Protozoan/bloodABSTRACT
Zoonotic cutaneous leishmaniasis (ZCL) is a neglected tropical disease caused by Leishmania (L.) major. This zoonosis is characterized by a broad-spectrum clinical polymorphism and may be underestimated and poorly treated since it is a simulator of various dermatoses. The aim of our study was to analyze the clinical polymorphism of patients with ZCL. A total of 142 patients with confirmed CL based on the microscopic examination of skin lesion biopsies were included in this study. Molecular typing of Leishmania species revealed that all patients were infected with L. major. In total, 14 clinical forms were observed. Six were typical and eight were atypical. The typical ZCL forms are grouped as follows: papular (26.76%), ulcero-crusted (26.05%), ulcerated (13.38%), impetiginous (9.86%), nodular (9.15%), and papulo-nodular (5.63%) lesions. In atypical ZCL forms, we described erythematous (2.81%), erysipeloid (1.4%), sporotrichoid, (1.4%), keratotic (0.7%) lupoid (0.7%), lichenoid (0.7%), psoriasiform (0.7%), and zosteriform (0.7%) lesions. Here, the lichenoid and the keratotic forms caused by L. major were reported for the first time in Tunisia. These findings will help physicians to be aware of the unusual lesions of ZCL that could be confused with other dermatological diseases. For this reason, it will be necessary to improve the diagnosis of CL especially in endemic areas. Such large clinical polymorphism caused by L. major may be the result of a complex association between the vector microbiota, the parasite, and the host immune state, and further studies should be carried out in order to reveal the mechanisms involved in clinical polymorphism of ZCL.
Subject(s)
Leishmaniasis, Cutaneous , Zoonoses , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/parasitology , Humans , Male , Female , Adult , Zoonoses/parasitology , Zoonoses/diagnosis , Middle Aged , Animals , Adolescent , Young Adult , Child , Leishmania major/genetics , Leishmania major/isolation & purification , Aged , Skin/parasitology , Skin/pathology , Child, PreschoolABSTRACT
BACKGROUND AND OBJECTIVE: Cutaneous leishmaniasis (CL) is still considered to be an uncontrolled endemic disease that spreads in many countries. The current study aimed to determine intra-species relationships of L. major using ITS2 sequencing. METHODS: The study was conducted from the beginning of March to the end of November 2022. All medical information regarding CL was collected from patients of Thi-Qar province who attended the Dermatology Department of Al-Hussein Teaching Hospital in Nasiriyah city. Seventy-three samples were selected for the molecular identification after confirming microscopy with Giemsa stain. In this study, the primers were designed using NCBI GenBank sequence database and Primer 3 plus primer design online software. RESULTS: The results recorded 21 (28.77%) positive samples of L. major using the internal transcribed spacer 2 region (ITS2) in ribosomal RNA gene. The local L. major IQN.1-IQN.10 were submitted to NCBI GenBank database with accession numbers OM069357.1-OM069366.1, respectively. The phylogenetic analysis revealed that local isolates of L. major showed a close relationship with NCBI-BLAST L. major Iran isolate (KU680848.1). CONCLUSION: ITS2-PCR is suitable for identifying Leishmania spp. and determining genetic diversity. A phylogenetic data analysis may provide an idea on the genetic homogeneity of local isolates and knowing the genetic origin of the dermal lesion. However, the local isolates showed genetic proximity to the KU680848.1 isolate. This signifies the possibility of infection prevalence from Iranian areas. In general, genetic variation of L. major isolates may give several clinical manifestations of the cutaneous lesion. Therefore, determination of the heterogeneity is important for detecting the infection origin, epidemiology, therapy, and control strategies.
Subject(s)
Genetic Variation , Leishmania major , Leishmaniasis, Cutaneous , Phylogeny , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Cutaneous/epidemiology , Leishmania major/genetics , Leishmania major/isolation & purification , Leishmania major/classification , Humans , DNA, Ribosomal Spacer/genetics , Male , Female , Iran/epidemiology , DNA, Protozoan/genetics , Adult , Middle Aged , Polymerase Chain Reaction , Adolescent , Child , Young Adult , Skin/parasitologyABSTRACT
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.
Subject(s)
Climate , Iran/epidemiology , Humans , Leishmaniasis/epidemiology , Leishmania/classification , Leishmania/isolation & purification , Leishmania infantum/isolation & purification , Geographic Information Systems , Temperature , Leishmania tropica/isolation & purification , Leishmania major/isolation & purificationABSTRACT
PURPOSE: Leishmania major is main causative agent and Phlebotomus papatasi is only proven vector of Zoonotic Cutaneous Leishmaniasis (ZCL) in Iran. Human leishmaniasis is mostly susceptible to climatic conditions and molecular variations of Leishmania parasites within sandflies. METHODS: L. major was analyzed based on geographical, environmental, climatic changes and haplotype variations within P. papatasi. Molecular tools and different geographical aspects were employed using Arc-GIS software for mapping the geographic distribution of samples and other statistics tests. Fragments of ITS-rDNA, k-DNA, and microsatellite genes of Leishmania were used for PCR, RFLP, sequencing, and phylogenetic analyses. RESULTS: Totally 81 out of 1083 female P. papatasi were detected with Leishmania parasites: 70 and five were L. major and L. turanica, respectively. Golestan and Fars provinces had the highest (13.64%) and lowest (4.55%) infection rates, respectively. The infection rate among female P. papatasi collected from gerbil burrows was significantly higher (15.15%) than animal shelters, yards, and inside houses (4.48%) (P < 0.0%). Microsatellite was more sensitive (22.72%) than k-DNA (18.8%) and ITS-rDNA (7.48%). More molecular variations of L. major were found in Isfahan province. CONCLUSIONS: Arc-GIS software and other statistics tests were employed to find Leishmania positive and haplotype variations among sand flies. Geographical situations, altitude, climate, precipitation, humidity, temperature, urbanization, migrations, regional divergences, deforestation, global warming, genome instability, ecology, and biology of the sand flies intrinsically, and the reservoir hosts and neighboring infected locations could be reasons for increasing or decreasing the rate of Leishmania infection and haplotype variations.
Subject(s)
Haplotypes , Leishmania major , Leishmaniasis, Cutaneous , Phlebotomus , Animals , Leishmania major/genetics , Leishmania major/isolation & purification , Phlebotomus/parasitology , Phlebotomus/genetics , Iran/epidemiology , Female , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Cutaneous/epidemiology , Leishmaniasis, Cutaneous/transmission , Phylogeny , Genetic Variation , Microsatellite Repeats , Insect Vectors/parasitology , Insect Vectors/genetics , DNA, Protozoan/genetics , Gerbillinae/parasitology , HumansABSTRACT
Objective: The relationship between drug resistance and the expression of hexokinase (HK) has been indicated in leishmaniasis. According to the prolonged treatment period in cutaneous leishmaniasis (CL) patients co-infected with Crithidia in Iran, this study aims to investigate the expression of HK in the proteome of Leishmania major and Crithidia using a proteomic approach. Methods: A total of 205 samples were removed from the lesions of patients in Fars province, Iran, for the characterization of L. major and Crithidia using polymerase chain reaction (PCR). After protein extraction, two-dimensional gel electrophoresis was employed for protein separation. Several spots were isolated for HK determination in the proteomes of L. major and Crithidia using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI TOF/TOF MS). Results: The PCR results showed 5 positive cases for Crithidia and 96 positive cases for L. major. MALDI TOF/TOF MS indicated HK as a common protein in the proteome of L. major and Crithidia. HK was up-regulated in the Crithidia proteome in comparison with the L. major proteome. Conclusion: Since a relationship between HK expression and drug resistance has been indicated in leishmaniasis, the overexpression of HK in Crithidia might be related to the increased duration of the treatment period in CL patients co-infected with Crithidia.
Subject(s)
Crithidia/metabolism , Hexokinase/metabolism , Leishmania major/metabolism , Proteome/metabolism , Coinfection/drug therapy , Coinfection/parasitology , Crithidia/enzymology , Crithidia/isolation & purification , Drug Resistance , Euglenozoa Infections/drug therapy , Euglenozoa Infections/parasitology , Humans , Iran , Leishmania major/enzymology , Leishmania major/isolation & purification , ProteomicsABSTRACT
BACKGROUND: Visceral leishmaniasis (VL) and zoonotic cutaneous leishmaniasis (ZCL) are of public health concern in Merti sub-County, Kenya, but epidemiological data on transmission, vector abundance, distribution, and reservoir hosts remain limited. To better understand the disease and inform control measures to reduce transmission, we investigated the abundance and distribution of sand fly species responsible for Leishmania transmission in the sub-County and their blood-meal hosts. METHODS: We conducted an entomological survey in five villages with reported cases of VL in Merti sub-County, Kenya, using CDC miniature light traps and castor oil sticky papers. Sand flies were dissected and identified to the species level using standard taxonomic keys and PCR analysis of the cytochrome c oxidase subunit 1 (cox1) gene. Leishmania parasites were detected and identified by PCR and sequencing of internal transcribed spacer 1 (ITS1) genes. Blood-meal sources of engorged females were identified by high-resolution melting analysis of vertebrate cytochrome b (cyt-b) gene PCR products. RESULTS: We sampled 526 sand flies consisting of 8 species, Phlebotomus orientalis (1.52%; n = 8), and 7 Sergentomyia spp. Sergentomyia squamipleuris was the most abundant sand fly species (78.71%; n = 414) followed by Sergentomyia clydei (10.46%; n = 55). Leishmania major, Leishmania donovani, and Trypanosoma DNA were detected in S. squamipleuris specimens. Humans were the main sources of sand fly blood meals. However, we also detected mixed blood meals; one S. squamipleuris specimen had fed on both human and mouse (Mus musculus) blood, while two Ph. orientalis specimens fed on human, hyrax (Procavia capensis), and mouse (Mus musculus) blood. CONCLUSIONS: Our findings implicate the potential involvement of S. squamipleuris in the transmission of Leishmania and question the dogma that human leishmaniases in the Old World are exclusively transmitted by sand flies of the Phlebotomus genus. The presence of Trypanosoma spp. may indicate mechanical transmission, whose efficiency should be investigated. Host preference analysis revealed the possibility of zoonotic transmission of leishmaniasis and other pathogens in the sub-County. Leishmania major and L. donovani are known to cause ZCL and VL, respectively. However, the reservoir status of the parasites is not uniform. Further studies are needed to determine the reservoir hosts of Leishmania spp. in the area.
Subject(s)
DNA, Protozoan/genetics , Leishmania donovani/genetics , Leishmania major/genetics , Leishmaniasis, Visceral/epidemiology , Psychodidae/parasitology , Trypanosoma/genetics , Animal Distribution , Animals , Blood/metabolism , DNA, Intergenic/genetics , Entomology/methods , Female , Humans , Hyraxes , Insect Vectors/parasitology , Kenya/epidemiology , Leishmania donovani/isolation & purification , Leishmania major/isolation & purification , Leishmaniasis, Visceral/prevention & control , Leishmaniasis, Visceral/transmission , Male , Meals , Mice , Psychodidae/classification , Psychodidae/genetics , Psychodidae/physiology , Trypanosoma/isolation & purificationABSTRACT
Immunomagnetic Separation (IMS) assay has been used for isolation of viable whole organisms. The objective of our work is to produce anti-Leishmania magnetic beads and to assess the efficiency of the IMS technique on Leishmania promastigote capture in culture media. Polyclonal anti-Leishmania antibodies were produced by intravenous injection of viable metacyclic promastigotes of Leishmania (L.) major to rabbit. Purified anti-Leishmania IgG was assessed for their reactivity against both L. major and L. infantum promastigotes then covalently conjugated to magnetic beads and used for IMS. This latter was applied on either L. major promastigote cultures of known concentrations or early stage (24h, 48h, 72h) Novy-MacNeal-Nicolle (NNN) cultures of tissue fluid obtained from cutaneous leishmaniasis (CL) lesions. Promastigotes capture was assessed by either microscopy or qPCR after sample boiling. Indirect immunofluorescence assay showed that polyclonal antibodies reacted against both L. major and L. infantum promastigotes. In 50 µL solution, immunomagnetic beads were able to capture 5 live promastigotes out of 20 and 1050 out of 2500, giving an estimated efficiency of 25-42%. The efficiency of the IMS was lower for a lower number of parasites but still repeatable. On the other hand, IMS-qPCR applied to 14 NNN cultures of confirmed Leishmania lesions showed a higher sensitivity to detect live parasites than routine microscopy observation of promastigotes growth (93% positivity at 72h versus 50% positivity within 2-4 weeks incubation). The estimated number of captured parasites at 72h ranged from 1 to more than 100 parasites / 50 µL liquid phase of culture. These preliminary results open the way for interesting perspectives in the use of cultures for leishmaniasis diagnosis and also for other applications such as Leishmania detection in cultures taken from reservoir animals or sandflies.
Subject(s)
Immunomagnetic Separation/methods , Leishmania infantum/isolation & purification , Leishmania major/isolation & purification , Animals , Female , Humans , Leishmania infantum/immunology , Leishmania major/immunology , RabbitsABSTRACT
Cutaneous leishmaniasis (CL) is transmitted by Phlebotomine sand fly vectors, among which Phlebotomus papatasi is prevalent in Western Asia, Northern Africa and Southern Europe, and it is known as a vector for Leishmania major parasite in the world. However, in Iraq, morphological studies showed that P. papatasi is a predominant sand fly species and hypothesised to transmit CL causing Leishmania species including L. major and L. tropica. Few studies have found Leishmania species in sand flies in mixed pools of samples in this country. Accurate identification of sand flies as vectors of Leishmania species is required in Iraq. The current study aims to identify sand fly species, using both morphological and molecular phylogenetic analyses, in a region where CL tends to be endemic. Furthermore, molecular phylogenetic analysis has also used to confirm Leishmania species in the sand fly samples collected in 11 villages between Diyala and Sulaymaniyah Provinces. For the first time, we have found L. major in three individual sand flies, one engorged (with fresh blood meal) and two non-engorged (without visible fresh blood meal) P. papatasi females in an area of CL outbreaks since 2014-till now due to civil wars and internal conflicts happen in the region. Further study should be performed on sand fly population and Leishmania reservoirs in this region.
Subject(s)
Disease Outbreaks , Insect Vectors/parasitology , Leishmania major/isolation & purification , Leishmaniasis, Cutaneous/epidemiology , Phlebotomus/parasitology , Animals , Female , Iraq/epidemiology , Leishmania major/genetics , Leishmaniasis, Cutaneous/parasitology , Male , PhylogenyABSTRACT
Visceral leishmaniasis is responsible for up to 30,000 deaths every year. Current treatments have shortcomings that include toxicity and variable efficacy across endemic regions. Previously, we reported the discovery of GNF6702, a selective inhibitor of the kinetoplastid proteasome, which cleared parasites in murine models of leishmaniasis, Chagas disease, and human African trypanosomiasis. Here, we describe the discovery and characterization of LXE408, a structurally related kinetoplastid-selective proteasome inhibitor currently in Phase 1 human clinical trials. Furthermore, we present high-resolution cryo-EM structures of the Leishmania tarentolae proteasome in complex with LXE408, which provides a compelling explanation for the noncompetitive mode of binding of this novel class of inhibitors of the kinetoplastid proteasome.
Subject(s)
Antiprotozoal Agents/chemistry , Antiprotozoal Agents/pharmacology , Leishmaniasis, Visceral/drug therapy , Oxazoles/chemistry , Oxazoles/pharmacology , Proteasome Inhibitors/chemistry , Proteasome Inhibitors/pharmacology , Pyrimidines/chemistry , Pyrimidines/pharmacology , Animals , Antiprotozoal Agents/therapeutic use , Dogs , Humans , Leishmania donovani/drug effects , Leishmania donovani/isolation & purification , Leishmania major/drug effects , Leishmania major/isolation & purification , Leishmaniasis, Visceral/parasitology , Liver/parasitology , Macaca fascicularis , Mice , Mice, Inbred BALB C , Oxazoles/therapeutic use , Proteasome Inhibitors/therapeutic use , Pyrimidines/therapeutic use , Rats , Rats, Sprague-Dawley , Triazoles/chemistryABSTRACT
Cutaneous leishmaniasis (CL) is an infectious disease caused by various Leishmania species. It is among the most neglected tropical diseases and has been considered a major health threat over the past decades in the country. Its zoonotic form caused by Leishmania (L) major is the most prevalent in Morocco. This study investigated the population structure of L. major in southeastern Morocco. Samples (n = 67) were collected from patients with CL in five different endemic areas located in three provinces (Ouarzazate, Tinghir, and Zagora). These samples were then sequenced using two nuclear markers: internal transcribed spacer 1 (ITS1) and a fragment of the virulence factor GP63. Next, the sequences were edited and analyzed. Molecular diversity indices showed a high population genetic diversity but an overall low haplotype diversity. Our results suggest small population differentiation, indicating a low geographic structure. Tajima's D and Fu's Fs tests both suggested recent population expansion based on the significant deviations from neutrality in both tests for all populations except Tinghir, which may be due to a small sample size. Based on our findings, the region is experiencing rapid population expansion caused by recent CL outbreaks, and one of them has been recently studied. In addition, analysis of molecular variance and FST suggested gene flow between Zagora and both Ouarzazate and Tinghir. Nonetheless, no gene flow was observed between Tinghir and Ouarzazate. To the best of our knowledge, this is the first analysis of the population structure of L. major in Morocco. The results of this study provide crucial background information for epidemiological studies by showing the presence of gene flow between populations and clonal expansion in cases of an outbreak. This will drive authorities to reconsider the implemented control strategies.
Subject(s)
Leishmania major/isolation & purification , Leishmaniasis, Cutaneous/parasitology , Animals , Haplotypes , Humans , Leishmania major/genetics , Morocco/epidemiologyABSTRACT
INTRODUCTION: in Burkina Faso, the only epidemic focus of cutaneous leishmaniasis confirmed in the literature by lab tests was in Ouagadougou. We report the epidemiological, clinical and biological results of the assessment of a new epidemic focus in Larama in western Burkina Faso. METHODS: camps were used to receive patients. Sociodemographic and clinical data were collected using a questionnaire. Confirmation was based on microscopy and polymerase chain reaction (PCR). RESULTS: a total of 108 suspected cases have been identified in Larama, reflecting an attack rate of 5.8%. Sex ratio was 1.08. The patients were most often farmers (35.2%) and traders (33.3%). The working population (15-49 years old) accounted for 51.9%. The number of lesions varied between 1 and 5 in 91.7% of the cases. The lesions manifested as raised and infiltrated ulcerative lesions on the limbs (87%) with evolution ranging from 1 to 5 months in 96.3% of the cases. Samples were collected from two patients; microscopy showed leishmanias and PCR confirmed Leishmania major. CONCLUSION: our results confirm the presence of a cutaneous leishmaniasis major outbreak in the western part of the country. Additional surveys are needed to clarify the burden of leishmaniasis in Burkina Faso.
Subject(s)
Disease Outbreaks , Leishmania major/isolation & purification , Leishmaniasis, Cutaneous/epidemiology , Adolescent , Adult , Burkina Faso/epidemiology , Child , Child, Preschool , Female , Humans , Infant , Male , Middle Aged , Polymerase Chain Reaction , Surveys and Questionnaires , Young AdultABSTRACT
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.
Subject(s)
Disease Reservoirs/veterinary , Gerbillinae , Leishmaniasis, Cutaneous/veterinary , Leishmaniasis, Visceral/veterinary , Murinae , Rodent Diseases/epidemiology , Animals , Female , Leishmania infantum/isolation & purification , Leishmania major/isolation & purification , Leishmania tropica/isolation & purification , Leishmaniasis, Cutaneous/epidemiology , Leishmaniasis, Cutaneous/parasitology , Leishmaniasis, Visceral/epidemiology , Leishmaniasis, Visceral/parasitology , Male , Prevalence , Rodent Diseases/parasitology , Turkey/epidemiologyABSTRACT
OBJECTIVE: Leishmaniasis is caused by different Leishmania spp. Treatment failure (TF) of cutaneous leishmaniasis (CL) is a serious issue that may be due to various reasons, previous studies suggested Leishmania RNA virus (LRV) as a potential cause of TF. Two variant groups of LRV1 and LRV2 are reported. In this study, the presence of LRV1/LRV2 was compared in TF with treatment response (TR) isolates of L. major. Clinical isolates of 15 TF and 15 TR were collected from CL patients referred to the Health Centers of Isfahan. Genomic DNA was extracted to identify Leishmania spp. using ITS1-PCR-RFLP. Identification of LRV1/LRV2 was performed using SYBR Green Real-Time PCR. The statistical analysis to test relationship between the treatment response with Glucantime and the presence of LRV were performed using SPSS 16.0 with Fisher's Exact test. P value of less than 0.05 was considered significant. RESULTS: ITS1-PCR-RFLP results showed that every isolate was identified as L. major. The results showed no LRV1 in any of the samples but 7 TR isolates and 2 TF isolates showed positive for LRV2. Statistical analysis showed no significant difference between the presence of LRV2 and response to Glucantime (p-value = 0.1086). Therefore, other mechanisms might be responsible for TF.
Subject(s)
Leishmania major/virology , Leishmaniasis, Cutaneous/virology , Leishmaniavirus/isolation & purification , Adult , Antiprotozoal Agents/therapeutic use , Female , Humans , Leishmania major/genetics , Leishmania major/isolation & purification , Leishmania major/pathogenicity , Leishmaniasis, Cutaneous/drug therapy , Leishmaniavirus/genetics , Male , Meglumine Antimoniate/therapeutic use , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , RNA, Viral/genetics , Sequence Analysis, DNA , Treatment FailureABSTRACT
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.
Subject(s)
Leishmania major/isolation & purification , Leishmania tropica/isolation & purification , Leishmaniasis, Cutaneous/epidemiology , Zoonoses/parasitology , Adult , Animals , Electrophoresis/methods , Enzyme-Linked Immunosorbent Assay , Humans , Incidence , Iran/epidemiology , Leishmania major/genetics , Leishmania tropica/genetics , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/parasitology , Polymerase Chain Reaction , Prevalence , Random Amplified Polymorphic DNA Technique , Zoonoses/epidemiologyABSTRACT
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.
Subject(s)
Genotyping Techniques , Leishmania infantum/isolation & purification , Leishmania tropica/isolation & purification , Leishmaniasis, Cutaneous/parasitology , Armed Conflicts , Azure Stains , Endemic Diseases , Female , Genotype , Humans , Leishmania infantum/classification , Leishmania infantum/genetics , Leishmania major/classification , Leishmania major/genetics , Leishmania major/isolation & purification , Leishmania tropica/classification , Leishmania tropica/genetics , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Cutaneous/epidemiology , Male , Real-Time Polymerase Chain Reaction , Syria/epidemiology , Syria/ethnology , Transients and Migrants , Turkey/epidemiologyABSTRACT
BACKGROUND & OBJECTIVES: Cutaneous leishmaniasis (CL) is still a public health threat in many rural areas of Iran. It causes considerable morbidity among a huge number of people in the endemic areas. Khuzestan is considered as a CL focus in Iran and recently several cases have been reported from Shush County. The aim of the current study was to determine different aspects of CL epidemiology in this County. METHODS: The study was conducted in four villages in the rural district of Shush County. In order to survey the scar and acute rate of the disease, 100 inhabitants were questioned in each village. In addition, to determine the reservoir host and vector of the disease, rodents were captured using Sherman traps, and sandflies were collected from around the rodent burrows using sticky paper traps. RESULTS: The scar and acute lesion rates were reported as 5.26 and 0.57% among inhabitants, respectively. Fifty-four rodents including Tatera indica (55.5%), Nesokia indica (33.3%), Mus musculus (9.3%) and Rattus norvegicus (1.9%) were captured and identified. Nested PCR showed 3 out of 54 rodents infected with Leishmania major. In total, 1122 sandflies were collected, and Leishmania major detected in Phlebotomus papatasi (4.35%), Sergentomyia dentata (23%), and Sergentomyia clydei (10%). INTERPRETATION & CONCLUSION: Zoonotic cutaneous leishmaniasis due to L. major prevailed in this area. The common and the main vector was Ph. papatasi and, T. indica seems to be the main while N. indica as a secondary reservoir host of the disease in this area.
Subject(s)
Insect Vectors/parasitology , Leishmania major/isolation & purification , Leishmaniasis, Cutaneous/epidemiology , Psychodidae/parasitology , Animals , Disease Reservoirs/parasitology , Female , Humans , Iran/epidemiology , Leishmania major/genetics , Leishmaniasis, Cutaneous/parasitology , Male , Mice , Muridae , Phlebotomus/parasitology , Public Health , Rats , ZoonosesABSTRACT
Three distinct noso-epidemiological cutaneous leishmaniasis (LC) entities coexist in Algeria: the so-called sporadic form of the North (LCN), the zoonotic form (LCZ) and the chronic form (LCC). The precise identification of the parasitic species involved in each of the forms makes it possible to specify the geographical distribution of each of the forms raised, to distinguish their clinical aspects, to guide the therapeutic behaviors and to adapt the control programs. Ninety-seven (97) human strains from 97 cases of LC were subjected to molecular characterization by PCR-ITS1 followed by sequencing of this inter-gene space. Our results confirm the endemicity of the three forms. The LCN, caused by L. infantum (17 isolates/97 i.e. 17.52%) is limited to the North of the country mainly (16 isolates/17). Its geographical distribution is superimposable to that of visceral leishmaniasis with an extension more and more reported in previously unaffected areas, such as the regions of Tlemcen and Oran in the West, Setif, Annaba and Collo in the East. The LCZ, due to L. major (70 strains/97 i.e. 72.16%), remains the dominant form in the arid and semi-arid zones (47 strains/70) with a progression towards the North (20/70 strains). Indeed, long confined to the Sahara, it shows a geographical extension outside its historic homes of Biskra and Abadla. This form is progressing dangerously towards the highlands and the steppe regions of the country. The most interesting fact was the identification of L. tropica for the first time in North-Central and North-West Algeria in Algerian patients who had never left the national territory. Out of the 10 strains of L. tropica identified, 8 belonged to patients of Syrian origin and 2 to Algerian patients. L. tropica was reported for the first time in 2008 in 6 patients living in Constantine (North-East Algeria) and in 2017, still in the North-East of the country, in Annaba. The observation of L. tropica in the North and Northeast center of the country, where L. infantum and L. major coexist, suggests changes in the epidemiology of cutaneous leishmaniasis in Algeria, which highlights the interest of more investigations to better understand the transmission cycle of the different entities.
Trois entités nosoépidémiologiques distinctes de leishmaniose cutanée (LC) coexistent en Algérie : la forme dite du nord, dite sporadique (LCN), la forme zoonotique (LCZ) et la forme chronique (LCC). L'identification précise des espèces parasitaires impliquées dans chacune des formes permet de préciser la répartition géographique de chacune de ces trois formes, de distinguer leurs aspects cliniques, d'orienter les conduites thérapeutiques et d'adapter les programmes de lutte. Quatre-vingt-dix-sept souches humaines ont fait l'objet d'une caractérisation moléculaire par une PCR-ITS1, suivie d'un séquençage de cet espace intergénique. Nos résultats confirment l'endémicité des trois formes. La LCN, causée par Leishmania infantum (17 isolats/97, soit 17,52 %), est limitée au nord du pays principalement (16 isolats/17). Sa répartition géographique est stable et superposable à celle de la leishmaniose viscérale. La LCZ, due à Leishmania major (70 souches/97, soit 72,16 %), reste la forme dominante au niveau des zones arides et semi-arides (47 souches/70) avec une progression vers le nord (20 souches/70). Cette forme connaît une extension géographique en dehors de ses foyers historiques et progresse vers les Hauts Plateaux et les régions steppiques du pays. Le fait remarquable est l'identification de Leishmania tropica pour la première fois au centre nord et nord-ouest de l'Algérie, chez des patients algériens n'ayant jamais quitté le territoire national. L'observation de L. tropica au centre nord et nord-ouest du pays, où coexistent L. infantum et L. major, suggère des modifications de l'épidémiologie de la LC en Algérie, ce qui souligne l'intérêt de plus d'investigations pour mieux comprendre le cycle de transmission des différentes entités.