Cause Of Cutaneous Leishmaniasis Research Articles

Detection of Cutaneouse Leishmaniasis species via PCR in Salah Adeen and Baghdad provences

Background: Leishmaniasis is a parasitic diseases that are spread worldwide due to various species of Leishmania, which are infect mammales diversity as well as human. L. tropica, L. major, and L. aethiopica which is common causes of cutaneous Leishmaniasis in Salah Adeen and Baghdad provences.
 Material and Methods: The present study was conducted to investigate the prevalence of cutaneous Leishmaniasis and to identify Leishmania parasites by using polymerase chain reaction (PCR) in some endemic areas of Iraq. A total of 117 samples of patients with suspected cutaneous Leishmaniasis were collected in different age groups. And both sexes 73 male and 44 female patients.
 Results: PCR results showed the percentage of infections 62.39% of males while 37.60% of females. The average age was 23.35 years (the range, from 1- 60 years), with the highest percentage of cases in the age group 1-4 years and the lowest rate in the age group (40-60 years). The highest infection was by L.tropica of L.major and lowest infection caused by L.aethiopica, where is considered first revealed in Iraq.
 Conclusion: The study found that males were more likely to be infected than females. The study revealed that polymerase chain reaction (PCR) is the most effective and sensitive method for detecting types of Cutaneous Leishmaniasis.

Detection of Cutaneouse Leishmaniasis species via PCR in Salah Adeen and Baghdad provences

Background: Leishmaniasis is a parasitic diseases that are spread worldwide due to various species of Leishmania, which are infect mammales diversity as well as human. L. tropica, L. major, and L. aethiopica which is common causes of cutaneous Leishmaniasis in Salah Adeen and Baghdad provences.
 Material and Methods: The present study was conducted to investigate the prevalence of cutaneous Leishmaniasis and to identify Leishmania parasites by using polymerase chain reaction (PCR) in some endemic areas of Iraq. A total of 117 samples of patients with suspected cutaneous Leishmaniasis were collected in different age groups. And both sexes 73 male and 44 female patients.
 Results: PCR results showed the percentage of infections 62.39% of males while 37.60% of females. The average age was 23.35 years (the range, from 1- 60 years), with the highest percentage of cases in the age group 1-4 years and the lowest rate in the age group (40-60 years). The highest infection was by L.tropica of L.major and lowest infection caused by L.aethiopica, where is considered first revealed in Iraq.
 Conclusion: The study found that males were more likely to be infected than females. The study revealed that polymerase chain reaction (PCR) is the most effective and sensitive method for detecting types of Cutaneous Leishmaniasis.
 
 http://dx.doi.org/10.25130/tjps.24.2019.009

Cloning and Recombinant Expression of Elongation Factor-1α of Leishmania mexicana.

Leishmania mexicana is an intracellular parasite that causes cutaneous leishmaniasis (CL) in some countries, including Mexico. Recently, we identified the elongation factor-1α (EF-1α) of L. mexicana by immunoproteomic analysis. In Leishmania donovani, this molecule has been reported as a virulence factor involved in downregulation of macrophages by no-canonical function when EF-1α interacts with protein tyrosine phosphatase-1 (SHP-1). However, in L. mexicana the key role of EF-1α in host-parasite relationship has not been elucidated, by this reason we started with cloning and recombinant expression of this antigen. A sequence of 1350 bp corresponding to EF-1α (EF-Lm) full-length gene was amplified from genomic DNA of L. mexicana (GenBank: MG256973); this gene contains only one nucleotide change: C464T, compared with L. mexicana reference sequence (GenBank: FR799570.1). The gene cloned (EF-Lm) codes for a protein of 449 residues. It was expressed in Escherichia coli and purified as 63 kDa sumo-fusion protein, which was recognized in the sera of patients diagnosed with CL. Our results show that EF-Lm is immunogenic during infection, and the rEF-Lm could be used for further analyses in the host-parasite relationship.

Molecular Modeling and Structural Stability of Wild-Type and Mutant CYP51 from Leishmania major: In Vitro and In Silico Analysis of a Laboratory Strain.

Cutaneous leishmaniasis is a neglected tropical disease and a major public health in the most countries. Leishmania major is the most common cause of cutaneous leishmaniasis. In the Leishmania parasites, sterol 14α-demethylase (CYP51), which is involved in the biosynthesis of sterols, has been identified as an attractive target for development of new therapeutic agents. In this study, the sequence and structure of CYP51 in a laboratory strain (MRHO/IR/75/ER) of L. major were determined and compared to the wild-type strain. The results showed 19 mutations including seven non-synonymous and 12 synonymous ones in the CYP51 sequence of strain MRHO/IR/75/ER. Importantly, an arginine to lysine substitution at position of 474 resulted in destabilization of CYP51 (ΔΔG = 1.17 kcal/mol) in the laboratory strain; however, when the overall effects of all substitutions were evaluated by 100 ns molecular dynamics simulation, the final structure did not show any significant changes (p-value < 0.05) in stability parameter of the strain MRHO/IR/75/ER compared to the wild-type protein. The energy level for the CYP51 of wild-type and MRHO/IR/75/ER strain were −40,027.1 and −39,706.48 Kcal/mol respectively. The overall Root-mean-square deviation (RMSD) deviation between two proteins was less than 1 Å throughout the simulation and Root-mean-square fluctuation (RMSF) plot also showed no substantial differences between amino acids fluctuation of the both protein. The results also showed that, these mutations were located on the protein periphery that neither interferes with protein folding nor with substrate/inhibitor binding. Therefore, L. major strain MRHO/IR/75/ER is suggested as a suitable laboratory model for studying biological role of CYP51 and inhibitory effects of sterol 14α-demethylase inhibitors.

Correction to: Molecular identification of Leishmania spp. isolates causes cutaneous leishmaniasis (CL) in Sanliurfa Province, Turkey, where CL is highly endemic.

In published article (Molecular Identification of Leishmania spp. Isolates Causes Cutaneous Leishmaniasis (CL) in Sanliurfa Province, Turkey, Where CL is Highly Endemic) Table 1 titled "The pH values of yogurts collected from villages in Turkey" doesn't belong to this article.

Evidence for Seroprevalence in Human Localized Cutaneous Leishmaniasis Caused by Leishmania donovani in Sri Lanka.

Visceral leishmaniasis (VL) is considered as a major health threat in the Indian subcontinent. Leishmania donovani, a usually visceralizing species, causes cutaneous leishmaniasis (CL) in Sri Lanka. However, visceralizing potential of the local L. donovani is not yet fully understood. This project studied the seroprevalence of local CL by using an in-house ELISA. An IgG-based ELISA using crude Leishmania antigen (Ag) was developed and validated. A total of 50 laboratory confirmed cases of locally acquired CL were examined using the newly developed ELISA. According to the optimized ELISA, seroprevalence of anti-Leishmania IgG antibodies in the study group was 34.0% (n = 17/50). Majority of seropositive individuals were males (n = 13/17), representing 76%. Nearly half of the seropositive individuals were young adults (20–40 years, n = 9/17, 53%). Higher proportions of single lesions, large lesions, and nodular lesions were associated with a seroconversion. A proportion of local L. donovani infections leading to CL have the ability to raise an antibody response in the host. This may indicate early systemic involvement as one possibility. Study of a large number of patients with adequate follow-up would be useful.

Molecular identification of Leishmania spp. isolates causes cutaneous leishmaniasis (CL) in Sanliurfa Province, Turkey, where CL is highly endemic.

Cutaneous leishmaniasis (CL) is an important public health problem in Turkey. CL has been most frequently seen in Sanliurfa. There is an expectation of increase in the population of leishmaniasis cases with the influence of Syrian refugees arriving in Turkey. In this study we aimed to diagnosis of CL and identifying of parasite from Leishmania isolates by using ITS 1 PCR RFLP. Samples were collected from 135 CL patients in Sanliurfa. After the specimens were inoculated in medium NNN, the ones which were cultures positive were cultivated in RPMI 1640 followed by PCR-RFLP. Genomic DNA was extracted phenol-chloroform procedure. Samples were examined by using ITS 1 PCR followed by RFLP analysis. Our results indicated that two species, L. tropica (132 samples) and L. major (3 samples), are responsible for cutaneous leishmaniasis in Sanlıurfa. Our study is the first scientific study in which it is reported molecular analyses of cutaneous leishmaniasis cases caused by L. major in Sanliurfa in Southestern Anatolia Region. Because CL cases caused by L.major are detected in our study, it is considered that genotyping is important for diagnosis of Leishmania and following change of epidemiology.

Efficacy, Safety and Cost-Effectiveness of Thermotherapy in the Treatment of Leishmania donovani-Induced Cutaneous Leishmaniasis: A Randomized Controlled Clinical Trial.

Leishmania donovani causes cutaneous leishmaniasis (CL) in Sri Lanka. Standard treatment is multiple, painful doses of intralesional sodium stibogluconate (IL-SSG). Treatment failures are increasingly reported, hence the need to investigate alternatives. Efficacy, safety, and cost-effectiveness of thermotherapy were assessed for the first time for L. donovani CL. A single blinded noninferiority randomized controlled trial was conducted on new laboratory-confirmed CL patients with single lesions (N = 213). Selected patients were randomly assigned to 1) test group (N = 98; single session of radiofrequency-induced heat therapy (RFHT) given at 50°C for 30 seconds) and 2) control group (N = 115; 1-3 mL IL-SSG given weekly, until cure/10 doses). Patients were followed-up fortnightly for 12 weeks to assess clinical cure. Cost of treatment was assessed using scenario building technique. Cure rates by 8, 10, and 12 weeks in RFHT group were 46.5%, 56.5%, and 65.9% as opposed to 28%, 40.8%, and 59.4% in IL-SSG group, with no major adverse events. Cure rate by RFHT was significantly higher at 8 weeks (P = 0.009, odds ratio [OR]: 2.236, confidence interval [CI]: 1.217-4.108) and 10 weeks (P = 0.035, OR: 1.881, CI: 1.044-3.388), but comparable thereafter. Cost of RFHT was 7 times less (USD = 1.54/patient) than IL-SSG (USD = 11.09/patient). A single application of RFHT is safe, cost-effective, and convenient, compared with multiple doses of IL-SSG in the treatment of L. donovani CL. Therefore, RFHT would be considered noninferior as per trial outcome when compared with standard IL-SSG therapy with multiple benefits for the patient and the national health care system.

Distribution pattern of anthroponotic cutaneous leishmaniasis caused by Leishmania tropica in Western Afghanistan during 2013-2014

Anthroponotic cutaneous leishmaniasis (ACL), caused by Leishmania tropica, is the main cause of cutaneous leishmaniasis (CL) in the Herat province, Western Afghanistan. We investigated the role of environmental factors on ACL distribution in Herat. Epidemiological data from 2457 patients were retrieved from the local WHO sub-office. Shapefile layers of districts, cities, villages, land cover, soil type and digital elevation model (DEM) of the Herat province were used to assess, by logistic regression modelling, the effects of land cover, soil types, elevation, and proximity to the Harirud river on the distribution of ACL.The key determinants of distribution were: (i) close proximity to the Harirud river, (ii) elevation between 700 and 1200m, (iii) intensive and intermittent irrigated cultivated land, and (iv) Haplocalcids with Torriorthents and Torrifluvents soil types. No ACL cases were found below 700m, and a few cases were present at >1200m in irrigated areas around the Harirud river. These findings suggest that moist soil and the humidity from irrigated areas found between 700 and 1200m provide suitable breeding sites of Phlebotomus sergenti, the main sandfly vector of L. tropica in Afghanistan. The effect of elevation also explains the predominance of ACL over ZCL in this region. The present study showed that distribution of ACL is strongly associated with environmental factors in West Afghanistan where the political and socio-economic conditions may also affect the epidemiology of CL.

Phenotypic and functional characteristics of HLA-DR+ neutrophils in Brazilians with cutaneous leishmaniasis.

The protozoan Leishmania braziliensis causes cutaneous leishmaniasis (CL) in endemic regions. In murine models, neutrophils (PMNs) are recruited to the site of infection soon after parasite inoculation. However, the roles of neutrophils during chronic infection and in human disease remain undefined. We hypothesized that neutrophils help maintain a systemic inflammatory state in subjects with CL. Lesion biopsies from all patients with CL tested contained neutrophils expressing HLA-DR, a molecule thought to be restricted to professional antigen-presenting cells. Although CL is a localized disease, a subset of patients with CL also had circulating neutrophils expressing HLA-DR and the costimulatory molecules CD80, CD86, and CD40. PMNs isolated from a low-density leukocyte blood fraction (LD-PMNs) contained a higher percentage of HLA-DR+ PMNs than did normal-density PMNs. In vitro coculture experiments suggested LD-PMNs do not suppress T cell responses, differentiating them from MDSCs. Flow-sorted HLA-DR+ PMNs morphologically resembled conventional PMNs, and they exhibited functional properties of PMNs. Compared with conventional PMNs, HLA-DR+ PMNs showed increased activation, degranulation, DHR123 oxidation, and phagocytic capacity. A few HLA-DR+ PMNs were observed in healthy subjects, and that proportion could be increased by incubation in either inflammatory cytokines or in plasma from a patient with CL. This was accompanied by an increase in PMN hladrb1 mRNA, suggesting a possible connection between neutrophil "priming" and up-regulation of HLA-DR. These data suggest that PMNs that are primed for activation and that also express surface markers of antigen-presenting cells emerge in the circulation and infected tissue lesions of patients with CL.

Cutaneous Leishmaniasis: The Truth about the ‘Flesh-Eating Disease’ in Syria

Recent news headlines claimed that corpses thrown into Syrian streets are causing cutaneous leishmaniasis (CL) outbreaks. However, leishmaniasis is only transmitted by blood-feeding sandflies, not through human remains. High CL prevalence in Syria may instead be attributed to the absence of disease control programs due to the disruption of health services.

Isolation and identification of cutaneous leishmaniasis species by PCR-RFLP in Ilam province, the west of Iran.

Cutaneous leishmaniasis (CL) is one of the most common parasitic diseases and public health problems in Iran. CL is endemic in most parts of Ilam province, in the west of Iran. The distance from the center of country, the great number of divers rural areas, and lack of specialists and laboratory facilities have been the major causes of Leishmania species remaining unknown in this region. Polymerase chain reaction followed by restriction fragment length polymorphism was performed to identify the Leishmania species in 61 patients with cutaneous lesions. Eventually L. major was confirmed as the cause of cutaneous leishmaniasis in Ilam province, the west of Iran.

Species-specific antimonial sensitivity in Leishmania is driven by post-transcriptional regulation of AQP1.

Leishmania is a digenetic protozoan parasite causing leishmaniasis in humans. The different clinical forms of leishmaniasis are caused by more than twenty species of Leishmania that are transmitted by nearly thirty species of phlebotomine sand flies. Pentavalent antimonials (such as Pentostam or Glucantime) are the first line drugs for treating leishmaniasis. Recent studies suggest that pentavalent antimony (Sb(V)) acts as a pro-drug, which is converted to the more active trivalent form (Sb(III)). However, sensitivity to trivalent antimony varies among different Leishmania species. In general, Leishmania species causing cutaneous leishmaniasis (CL) are more sensitive to Sb(III) than the species responsible for visceral leishmaniasis (VL). Leishmania aquaglyceroporin (AQP1) facilitates the adventitious passage of antimonite down a concentration gradient. In this study, we show that Leishmania species causing CL accumulate more antimonite, and therefore exhibit higher sensitivity to antimonials, than the species responsible for VL. This species-specific differential sensitivity to antimonite is directly proportional to the expression levels of AQP1 mRNA. We show that the stability of AQP1 mRNA in different Leishmania species is regulated by their respective 3’-untranslated regions. The differential regulation of AQP1 mRNA explains the distinct antimonial sensitivity of each species.

Identification of Cutaneous Leishmaniasis Agents in Four Geographical Regions of Khuzestan Province Using Nested PCR

Background: Leishmania tropica and Leishmania major are the main causes of cutaneous leishmaniasis in endemic regions of Iran. Objectives: The aim of this study was the identification of cutaneous leishmaniasis agents in Khuzestan province located southwest of Iran. Patients and Methods: 146 samples were collected from the lesions of 146 individuals including 67 (59.59% ) male and 59 ( 40.41% ) female with cutaneous leishmaniasis. The samples were then delivered to Iran- Zamin diagnostic laboratory and smeared on slides, stained with Wright’s eosin methylene blue stain and examined microscopically and graded from 1+ to 4+. DNA was extracted from the slides and the identification of cutaneous leishmaniasis agents was performed using Nested PCR with the primers of CSB1XR, CSB2XF, LIR and 13Z. Results: 138 (94.5%) out of 146 cases of four regions were L. major and 8 (5.5%) were L. tropica. 57.97% of L. major cases were male and 42.03% were female. 87.5% of L. tropica were male and 12.5% were female. The maximum number of L. tropica cases was found in the northern region (8.16%) and the minimum was found in the western region (3.22%). 96.78% of L. major cases belonged to the western region of Khuzestan. Conclusions: L. major is the main species responsible for cutaneous leishmsniasis in four geographical regions of Khuzestan province southwestern of Iran and Nested PCR can be used for diagnosis and Leishmania species identification.

Leishmania donovani zymodeme MON-37 isolated from an autochthonous visceral leishmaniasis patient in Sri Lanka

Although the strain causing cutaneous leishmaniasis (CL) in Sri Lanka was first identified in 2003, the strain causing visceral leishmaniasis (VL) has not yet been identified. We report the first isoenzyme typing of a strain causing VL in Sri Lanka at an early stage of emergence of VL in the country. The parasite was isolated from a 57-year-old civil soldier who had been in the jungle in the Vavuniya district in the Northern Province of Sri Lanka for a period of nearly 6 months immediately before the onset of symptoms. Multilocus enzyme electrophoresis (MLEE) revealed that the strain is Leishmania donovani zymodeme MON-37, the zymodeme which was previously identified from the CL patients in the country. The MLEE analysis was confirmed by sequencing the gene encoding the 6-phosphogluconate dehydrogenase isoenzyme. This is an instance of the same Leishmania zymodeme associated with both dermotropism and viscerotropism in the same geographic region. Further investigations into the genetic structure and identification of virulence factors in the parasite and immune factors in the host are required to understand the factors responsible for different tropism shown by the same zymodeme MON-37 L. donovani from Sri Lanka.

Molecular epidemiology of cutaneous leishmaniasis and heterogeneity of Leishmania major strains in Iran

To determine the geographical distribution of Leishmania species causing cutaneous leishmaniasis (CL) and to study the genetic heterogeneity of Leishmania major isolates from different endemic areas of Iran. A total of 341 isolates from lesions of patients living in 11 provinces of Iran were grown in culture medium and inoculated to BALB/c mice to detect possible visceralisation. The species were identified by isoenzyme analysis using a battery of six enzymes and kinetoplast (k) DNA-PCR technique. Genetic variation among L. major isolates was analysed by random amplified polymorphic DNA (RAPD) technique. Of the total 341 isolates, 283 isolates were L. major and 58 isolates were Leishmania tropica. In rural areas, the causative agent of CL was mainly L. major (95%L. major vs. 5%L. tropica), in urban areas it was L. tropica (65%L. tropica vs. 35%L. major). All isolates of L. major and 8.6% of L. tropica isolates showed visceralisation in BALB/c mice. There is considerable genetic diversity between L. major strains from different endemic areas and even between some isolates of the same endemic area. Leishmania major is the most frequent species in the endemic areas of CL in eleven provinces of Iran, and genetic diversity is a common feature of L. major in the country.

AutochthonousLeishmania siamensisin Horse, Florida, USA

Autochthonous<i>Leishmania siamensis</i>in Horse, Florida, USA

Methods incorporating a polymerase chain reaction and restriction fragment length polymorphism and their use as a ‘gold standard’ in diagnosing Old World cutaneous leishmaniasis

Three polymerase chain reaction (PCR) assays — kinetoplast DNA (kDNA) PCR, 7SL PCR, ITS1 PCR — were compared for their ability to detect leishmanial parasites in the skin tissue aspirates of 212 Palestinian suspect cases of cutaneous leishmaniasis (CL). The kDNA PCR was the most sensitive, detecting 156/170 (91.8%) of positive samples confirmed by a set ‘gold standard’ but of poor specificity in identifying the species of Leishmania. The 7SL PCR detected 154/170 (90.5%) and the ITS1 PCR only 108/170 (63.5%) of the true positives, and both were 100% specific. The highest Kappa coefficient agreement (95% CI) was obtained with the 7SL PCR (0.792 ± 0.098). Restriction analysis of the products from the ITS1 PCR and 7SL PCR enabled identification of species of Leishmania. L. tropica was the predominant cause of CL, with L. major being less frequent.

Identification of geographically distributed sub-populations of Leishmania (Leishmania) major by microsatellite analysis.

BackgroundLeishmania (Leishmania) major, one of the agents causing cutaneous leishmaniasis (CL) in humans, is widely distributed in the Old World where different species of wild rodent and phlebotomine sand fly serve as animal reservoir hosts and vectors, respectively. Despite this, strains of L. (L.) major isolated from many different sources over many years have proved to be relatively uniform. To investigate the population structure of the species highly polymorphic microsatellite markers were employed for greater discrimination among it's otherwise closely related strains, an approach applied successfully to other species of Leishmania.ResultsMultilocus Microsatellite Typing (MLMT) based on 10 different microsatellite markers was applied to 106 strains of L. (L.) major from different regions where it is endemic. On applying a Bayesian model-based approach, three main populations were identified, corresponding to three separate geographical regions: Central Asia (CA); the Middle East (ME); and Africa (AF). This was congruent with phylogenetic reconstructions based on genetic distances. Re-analysis separated each of the populations into two sub-populations. The two African sub-populations did not correlate well with strains' geographical origin. Strains falling into the sub-populations CA and ME did mostly group according to their place of isolation although some anomalies were seen, probably, owing to human migration.ConclusionThe model- and distance-based analyses of the microsatellite data exposed three main populations of L. (L.) major, Central Asia, the Middle East and Africa, each of which separated into two sub-populations. This probably correlates with the different species of rodent host.

Identification of Leishmania donovani as a cause of cutaneous leishmaniasis in Sudan

Eight patients with cutaneous ulcers were referred to the Institute of Endemic Diseases, Khartoum, Sudan, from June 2000 to March 2002 for the diagnosis of suspected cutaneous leishmaniasis (CL). Diagnosis was confirmed parasitologically by both positive Giemsa-stained smears and successful culture of Leishmania promastigotes in NNN medium. The eight parasite isolates were shown to belong to the Leishmania donovani complex by kDNA PCR. Isoenzyme typing of three isolates revealed that they were identical to the L. donovani MON-82 reference strain, and the gp63 PCR-RFLP profile showed similar patterns to a reference strain of MON-82. CL is endemic in most regions of Sudan and has been reported previously as being caused by L. major MON-74. The results of this study suggest that L. donovani is also a cause of CL in Sudan and that further study of isolates from Sudanese patients with cutaneous ulcers is warranted to ascertain whether L. donovani or L. major is the causative agent.