Cause Of Cutaneous Leishmaniasis Research Articles

Salivary antigens rPagSP02 and rPagSP06 are a reliable composite biomarker for evaluating exposure to Phlebotomus argentipes in Sri Lanka

Phlebotomus argentipes is the established vector of leishmaniasis in the Indian sub-continent. Antibodies to sand fly salivary antigens are biomarkers for vector-host exposure in leishmaniasis-endemic regions. Ph. argentipes transmits Leishmania donovani in Sri Lanka, primarily causing cutaneous leishmaniasis (CL). Our study compared the performance of salivary gland homogenate (SGH) from a lab-reared local strain of Ph. argentipes females to a composite recombinant salivary biomarker (rPagSP02 + rPagSP06) in a CL-endemic population. Sera from 546 healthy individuals, 30 CL patients, and 15 non-endemic individuals were collected. Western blot analysis of Ph. argentipes SGH identified immunogenic bands between 15 kDa and 67 kDa, with bands of predicted molecular weight ∼of 15 kDa (SP02) and ∼28–30 kDa (SP06) as the major antibody targets. Indirect ELISAs using SGH or rPagSP02 + rPagSP06 antigens showed high sensitivity (96.7%) and specificity (100%), detecting comparable seropositivity in endemic populations. rPagSP02 + rPagSP06 exhibited enhanced discriminatory ability, supported by a strong positive correlation (r = 0.869) with SGH. Our findings indicate that the composite rPagSP02 + rPagSP06 salivary biomarker effectively identifies Ph. argentipes exposure in individuals living in Sri Lanka, showing promising potential for use in surveillance. These findings should be further validated to confirm the epidemiological applications in leishmaniasis-endemic regions.

Autochthonous Leishmaniasis Caused by Leishmania tropica, Identified by Using Whole-Genome Sequencing, Sri Lanka.

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.

RPagSP02+rPagSP06 recombinant salivary antigen is a reliable biomarker for evaluating exposure to Phlebotomus argentipes in Sri Lanka.

Phlebotomus argentipes is the established vector of leishmaniasis in the Indian sub-continent. Antibodies to sand fly salivary antigens are biomarkers for vector-host exposure in leishmaniasis-endemic regions. Ph. argentipes transmits Leishmania donovani in Sri Lanka, primarily causing cutaneous leishmaniasis (CL). Our study compared the performance of salivary gland homogenate (SGH) from a lab-reared local strain of Ph. argentipes females to a composite recombinant salivary biomarker (rPagSP02 + rPagSP06) in a CL-endemic population. Sera from 546 healthy individuals, 30 CL patients, and 15 non-endemic individuals were collected. Western blot analysis of Ph. argentipes SGH identified immunogenic bands between 15 kDa and 67 kDa, with bands of predicted molecular weight ∼of 15 kDa (SP02) and ∼28-30 kDa (SP06) as the major antibody targets. Indirect ELISAs using SGH or rPagSP02 + rPagSP06 antigens showed high sensitivity (96.7%) and specificity (100%), detecting comparable seropositivity in endemic populations. rPagSP02 + rPagSP06 exhibited enhanced discriminatory ability, supported by a strong positive correlation (r = 0.869) with SGH. Our findings indicate that the composite rPagSP02 + rPagSP06 salivary biomarker effectively identifies Ph. argentipes exposure in individuals living in Sri Lanka, showing promising potential for use in surveillance. These findings should be further validated to confirm the epidemiological applications in leishmaniasis-endemic regions.

Molecular Characterization of Agents of Cutaneous Leishmaniasis in Dutsin-Ma Local Government Area of Katsina State, Northwestern, Nigeria

Study’s Novelty/Excerpt This study identified Leishmania major as the predominant species causing cutaneous leishmaniasis (CL) in Dutsin-ma LGA, Katsina state, Nigeria, using molecular characterization techniques such as ITS rDNA gene amplification and RFLP analysis. The findings revealed that 85.7% of individuals with suspected CL lesions were confirmed to have Leishmania infections, with a higher prevalence in males (71.4%) and farmers (71.46%). These results highlight the endemic nature of CL in this region, emphasizing the need for targeted public health interventions and awareness programs to control the spread of this neglected tropical disease. Full Abstract Leishmaniasis is one of man's neglected tropical diseases (NTD), caused by a protozoan parasite of the Leishmania genus. The dixenous life cycle of Leishmania species consists of the vertebrate and vector stages resulting in the differentiation of metabolic processes, morphological forms, and remodelling of genes in the parasite. To identify the species of Leishmania responsible for cutaneous leishmaniasis (CL) in Dutsin-ma LGA of Katsina state, Nigeria, a purposive sampling technique was employed. Samples were collected from individuals with suspected CL lesions and subjected to molecular characterization to confirm the Leishmania species. Questionnaires were also used to gather socio-demographic and behavioral data. Molecular characterization involved amplifying the ITS rDNA genes and applying the restriction fragment length polymorphism (RFLP) technique. The results revealed that 85.7% of the individuals screened had lesions associated with Leishmania infection, with a higher prevalence in males (71.4%) compared to females (14.3%). Among the infected individuals, 71.46% were farmers, and one was a student. The lesions were predominantly found on the legs (57.1%) and hand/arm (28.6%), with no lesions detected on other body parts. The ITS rDNA genes were successfully amplified from six patients, confirming Leishmania major as the predominant species causing CL through RFLP analysis. In conclusion, CL caused by L. major is endemic in Dutsin-ma LGA, Katsina state, in the Northwestern zone of Nigeria

Piperine, quercetin, and curcumin identified as promising natural products for topical treatment of cutaneous leishmaniasis

Leishmania braziliensis (L. braziliensis) causes cutaneous leishmaniasis (CL) in the New World. The costs and the side effects of current treatments render imperative the development of new therapies that are affordable and easy to administer. Topical treatment would be the ideal option for the treatment of CL. This underscores the urgent need for affordable and effective treatments, with natural compounds being explored as potential solutions. The alkaloid piperine (PIP), the polyphenol curcumin (CUR), and the flavonoid quercetin (QUE), known for their diverse biological properties, are promising candidates to address these parasitic diseases. Initially, the in vitro cytotoxicity activity of the compounds was evaluated using U-937 cells, followed by the assessment of the leishmanicidal activity of these compounds against amastigotes of L. braziliensis. Subsequently, a golden hamster model with stationary-phase L. braziliensis promastigote infections was employed. Once the ulcer appeared, hamsters were treated with QUE, PIP, or CUR formulations and compared to the control group treated with meglumine antimoniate administered intralesionally. We observed that the three organic compounds showed high in vitro leishmanicidal activity with effective concentrations of less than 50 mM, with PIP having the highest activity at a concentration of 8 mM. None of the compounds showed cytotoxic activity for U937 macrophages with values between 500 and 700 mM. In vivo, topical treatment with QUE daily for 15 days produced cured in 100% of hamsters while the effectiveness of CUR and PIP was 83% and 67%, respectively. No failures were observed with QUE. Collectively, our data suggest that topical formulations mainly for QUE but also for CUR and PIP could be a promising topical treatment for CL. Not only the ease of obtaining or synthesizing the organic compounds evaluated in this work but also their commercial availability eliminates one of the most important barriers or bottlenecks in drug development, thus facilitating the roadmap for the development of a topical drug for the management of CL caused by L. braziliensis.

Treatment outcomes of cutaneous leishmaniasis due to Leishmania aethiopica: A systematic review and meta-analysis.

Leishmania aethiopica is a unique species that causes cutaneous leishmaniasis (CL), and studies evaluating treatment outcomes for this condition reported inconsistent findings. This study aimed to summarize the evidence on treatment outcomes of CL caused by L. aethiopica to support decisions or propose further study. We searched PubMed, Scopus, and ScienceDirect. In addition, we searched grey literature on Google Scholar and performed manual searching on the reference list of articles. Two authors did the screening, selection, critical appraisal, and data extraction. With the narrative synthesis of evidence, we performed a random effects model meta-analysis using the metaprop package in Stata 17. We did sensitivity and subgroup analyses after assessing heterogeneity using the I-squared test and forest plots. The funnel plot and Egger's test were used to assess publication bias. The review included 22 studies with 808 participants, and the meta-analysis included seven studies with 677 participants. Most studies documented treatment outcomes with antimonial monotherapy, and only one study reported outcomes with combination therapy. The overall pooled proportion of cure was 63% (95% CI: 38-86%). In the subgroup analysis, systemic antimonial monotherapy showed a cure rate of 61%, and the proportion of cure was 87% with topical therapy. Topical therapy showed a better cure for the localized clinical phenotype. A cohort study documented a cure rate of 94.8% with combination therapy for the localized, mucocutaneous, and diffuse clinical phenotypes. The pooled proportion of unfavourable outcomes was partial response (19%), relapse (17%), discontinuation (19%), and unresponsiveness (6%). The pooled proportion of cure is low with antimonial monotherapy. Despite limited evidence, combination therapies are a promising treatment option for all clinical phenotypes of CL caused by L. aethiopica. Future high-quality randomized control trials are needed to identify effective monotherapies and evaluate the effectiveness of combination therapies.

Comparative Proteomics and Genome-Wide Druggability Analyses Prioritized Promising Therapeutic Targets against Drug-Resistant Leishmania tropica

Leishmania tropica is a tropical parasite causing cutaneous leishmaniasis (CL) in humans. Leishmaniasis is a serious public health threat, affecting an estimated 350 million people in 98 countries. The global rise in antileishmanial drug resistance has triggered the need to explore novel therapeutic strategies against this parasite. In the present study, we utilized the recently available multidrug resistant L. tropica strain proteome data repository to identify alternative therapeutic drug targets based on comparative subtractive proteomic and druggability analyses. Additionally, small drug-like compounds were scanned against novel targets based on virtual screening and ADME profiling. The analysis unveiled 496 essential cellular proteins of L. tropica that were nonhomologous to the human proteome set. The druggability analyses prioritized nine parasite-specific druggable proteins essential for the parasite's basic cellular survival, growth, and virulence. These prioritized proteins were identified to have appropriate binding pockets to anchor small drug-like compounds. Among these, UDPase and PCNA were prioritized as the top-ranked druggable proteins. The pharmacophore-based virtual screening and ADME profiling predicted MolPort-000-730-162 and MolPort-020-232-354 as the top hit drug-like compounds from the Pharmit resource to inhibit L. tropica UDPase and PCNA, respectively. The alternative drug targets and drug-like molecules predicted in the current study lay the groundwork for developing novel antileishmanial therapies.

Cutaenous Leishmaniasis In Libya

Cutaneous Leishmaniasis is wide spread in northwestern of Libya with increasing of incidence in the last years, as well as with appearance of new foci in Tawarga, Sirte, and Zliten, to become also endemic areas, and added to old known endemic areas like Jabal Nafusa. Different clinical manifestations and atypical presentations of skin lesions necessitate more clinical alertness as well as use of laboratory diagnostic procedures. Leishmania major was the main cause of cutaneous Leishmaniasis in Libya, followed by L. tropica, which was less frequent cause in less than one-third of all molecularly investigated patients. Some of main used therapies in Libya were cryotherapy, and intralesional antimony for patients with few and small lesions, although, in some patients; with large number as well as large size of lesions or patients with appearance of lesions at face or ears; made systemic treatment with sodium stibogluconate or meglumine antimoniate more suitable and mandatory treatment. In Libya, low doses of antimony as intramuscular injection were the followed regime that was recommended by national guidelines. Oral rifampicin and isoniazid were also successful alternative systemic therapy when systemic antimony was unsuitable or not effective.

Genotypic and phylogenic analyses of cutaneous leishmaniasis in Al Ahsa, Eastern Saudi Arabia during the coronavirus disease 2019 pandemic: First cases of Leishmania tropica with the predominance of Leishmania major

During the coronavirus disease 2019 lockdown period, a surge in sandflies and cutaneous leishmaniasis (CL) cases was observed in Al-Ahsa, Saudi Arabia. Skin punch biopsies were obtained from 100 patients clinically diagnosed with CL in Al-Ahsa who had no travel history in the last 6 months. Impression smears were used following a three-step polymerase chain reaction (PCR) protocol using genus-specific primers targeting kDNA and ITS1. Leishmania speciation was determined by ITS1 PCR/nested PCR-restriction fragment length polymorphism and sequencing. A phylogenetic tree was constructed. The associated patient characteristics were analyzed. Using internal transcribed spacer one (ITS1)-PCR/nested PCR, 98 cases were considered true-positive CL. Leishmania major was the predominant species, and Leishmania tropica was identified in three cases. Microscopy had poor sensitivity and perfect specificity. Direct ITS1-PCR missed nine cases. Sex, residence, and treatment outcome were significantly associated with the occurrence of Leishmania; distribution of skin lesion(s) and treatment outcome were significantly associated with Leishmania genotype. This is the first time that L. tropica was identified as a cause of CL in human in Al-Ahsa, in addition to the predominant zoonotic species, L. major. We recommend using ITS1-nested PCR for negative cases by ITS1-PCR. Further exploration of Leishmania transmission dynamics in vectors and reservoir animals is essential for designing effective preventive measures.

Prevalence of Leishmania RNA virus in Leishmania parasites in patients with tegumentary leishmaniasis: A systematic review and meta-analysis.

Cutaneous leishmaniasis is caused by different protozoan parasites of the genus Leishmania. Leishmania RNA virus (LRV) was identified as the first Leishmania infecting virus in 1998. Different studies showed the presence and role of the LRV in Leishmania parasites causing cutaneous leishmaniasis (CL). However, there is limited data on the pooled prevalence of LRV in Leishmania parasites causing CL. Therefore, the aim of this systematic review and meta-analysis was to determine the pooled prevalence of LRV in Leishmania parasite isolates and/or lesion biopsies in patients with CL from the available literature globally. We retrieved the studies from different electronic databases. The studies were screened and identified based on the inclusion and exclusion criteria. We excluded studies exclusively done in experimental animals and in vitro studies. The review was conducted in line with PRISMA guidelines. The meta-analysis was performed with Stata software version 14 with metan command. The forest plot with random-effect model was used to estimate the pooled prevalence with 95% confidence interval. Inverse variance index (I2) was used to assess the heterogeneity among the included articles. A total of 1215 samples from 25 studies were included. Of these, 40.1% (487/1215) were positive for LRV. The overall pooled prevalence of LRV globally was 37.22% (95% CI: 27.54% - 46.90%). The pooled prevalence of LRV in the New World (NW) and Old World (OW) regions was 34.18% and 45.77%, respectively. Leishmania guyanensis, L. braziliensis, L. major, and L. tropica were the most studied species for the detection of LRV. The prevalence of LRV from Leishmania isolates and lesion biopsies was 42.9% (349/813) and 34.3% (138/402), respectively. This systematic study revealed that there is high prevalence of LRV in Leishmania parasites isolated from patients with CL. More comprehensive studies would be required to investigate the presence of the LRV in other Leishmania species such as L. aethiopica to fully understand the role of LRV in different clinical manifestations and disease pathology presented in CL patients.

Antibody Responses to Phlebotomus papatasi Saliva in American Soldiers With Cutaneous Leishmaniasis Versus Controls

Leishmania major, transmitted in Iraq by the bite of a sand fly Phlebotomus papatasi, causes cutaneous leishmaniasis (CL). The sand fly saliva is immunogenic, with both systemic humoral and cellular human immune responses resulting from natural exposure. 248 Americans who developed L. major infection in Iraq were sex, race/ethnicity, year of Iraq deployment-matched to controls without CL. Using a case-control study design, we compared sand fly saliva-specific human IgG levels and recognized antigens between the two groups. Serologic responses to Ph. papatasi salivary gland homogenate were studied with ELISA and Western blot, using serial samples obtained from before travel, during CL treatment (CL) or at time of return to US (controls), as well as (for CL cases) six to 24 months after return to non-endemic US. The mean change in optical density (MCOD), reflecting the change in sand fly saliva-specific IgG before and after exposure in Iraq, was 0.296 (range -0.138 to 2.057) in cases and 0.151 (range -0.454 to1.085) in controls, p<0.001. Low levels of sand fly saliva specific antibody were noted in CL cases by 7-8 months after return to the US. The most frequently recognized Ph. papatasi salivary antigens were MW30 (PpSP32) and MW64, although other salivary proteins recognized were MW12/14, 15, 18, 28, 32, 36, 42, 44, 46, 52. Logistic regression suggested that MW15, 28 and 42 were associated with the largest effect on the MCOD. MW30 was the most frequently recognized antigen suggesting a role as biomarker for sand fly exposure and CL risk. Anti-Ph. papatasi saliva IgG waned within months of return to the US. We also discuss vector antigenic saliva proteins in the context of CL presentation and identify some salivary antigens that may correlate with less lesion area, ulcer versus papule/plaque, race among those with CL.

Distinct microbiome profiles and biofilms in Leishmania donovani-driven cutaneous leishmaniasis wounds

The endemic strain of Leishmania donovani in Sri Lanka causes cutaneous leishmaniasis (CL) rather than more common visceral form. We have visualized biofilms and profiled the microbiome of lesions and unaffected skin in thirty-nine CL patients. Twenty-four lesions (61.5%) were biofilm-positive according to fluorescence in situ hybridization. Biopsies of biofilm-positive lesions were dominated by Pseudomonas, class Bacilli and Enterobacteriaceae and distinguished by significantly lower community evenness. Higher relative abundance of a class Bacilli OTU was detected in wound swabs versus contralateral skin. Wound swabs and biopsies had significantly distinct microbiome profiles and lower diversity compared to unaffected skin. Greater abundances of potentially pathogenic organisms were observed in wet ulcers, lesions with high parasite loads and large wounds. In summary, more than half of L. donovani associated CL wounds harboured biofilms and the wounds exhibited a distinct, less diverse, microbiome than unaffected skin.

Detecting the presence of GRP-78 gene in syrian strain of leishmania tropica genome and sequencing it

Background: Leishmania tropica is the main cause of cutaneous leishmaniasis which represents a common issue in Syria. After the start of the current war, cutaneous leishmaniasis has become yet more widespread, so an effective vaccine is needed to prevent a largescale spread of leishmania in the country. DNA vaccination is a promising approach for developing such needed vaccine against leishmaniasis. Glucose-regulated protein 78 plays important roles in Leishmania parasite. The gene that codes this protein had been used as DNA vaccine against infections by different species of Leishmania, so this gene represents a good candidate for DNA vaccine designing. But the existence of glucose-regulated protein 78 gene has not yet been proven in the genome of Leishmania tropica. So, this study aims to detect the existence and the expression of glucoseregulated protein 78 gene in Syrian strain of Leishmania tropica promastigotes. Materials & methods: The DNA and RNA were isolated from Syrian strain of Leishmania tropica parasites, the cDNA was synthesized, necessary primers for genes’ amplification were designed manually, the PCR conditions were optimized. Full-length double-stranded sequence analysis was performed on PCR products from cDNA. Results: Extracted genomic DNA and total RNA showed high degree of purity and integrity. GRP78 PCR products showed by gel electrophoresis, on the level of DNA and cDNA, only one band belongs to the gene, and their sizes were approximately 1977 bp. The sequence of the cDNA-GRP78 gene was determined and published in GenBank, and the precise size of the gene was 1977 bp. Expression was also demonstrated at the level of cDNA. Conclusion: Glucose-regulated protein 78 gene is a part of Syrian strain of L. tropica, the expression of this gene has been demonstrated. The sequence of the cDNA of glucose-regulated protein 78 gene has defined and submitted to the Genbank with accession number NO. MN458469

Influence of Intestinal Helminth Burden on Clinical Manifestations, Therapeutic Response, and Leishmania braziliensis Load in Patients with New World Cutaneous Leishmaniasis.

Leishmania braziliensis is the most important cause of cutaneous leishmaniasis (CL) in the Americas. A Th1-type immune response is required to control Leishmania infection, but an exaggerated inflammatory response leads to the development of ulcers seen in CL. Infection with intestinal helminths has the potential to inhibit the Th1 response in a manner that depends both on the species of helminth present as well as the burden of helminthiasis. We conducted a prospective cohort study of CL patients from an endemic area between January and December 2017 with either negative or high intestinal helminth burden to characterize relationships between helminth burden, L. braziliensis quantification within CL lesions, clinical aspects of CL, and therapeutic response. Of 234 participants with leishmaniasis who underwent stool examination at the time of diagnosis, 45% had detectable helminth infection. The overall cure rate after 90 days was 66%, with a median time to resolution of disease of 40 days (interquartile range: 30-65 days). There was no significant association between the type of helminth infection or the magnitude of intestinal helminth burden at the time of diagnosis and L. braziliensis genomic DNA (gDNA) detected in biopsies from CL lesions. Likewise, there was no association between helminth burden and response to treatment after 90 days. Considering quantification of parasite DNA in CL lesions, participants who were cured at 90 days had a median of 0.017 ng/mg gDNA, and participants who failed therapy had a median of 0.091 ng/mg gDNA (P = 0.03). The results indicate that cutaneous Leishmania load may influence therapeutic response in CL.

A New Target Organ of Leishmania (Viannia) braziliensis Chronic Infection: The Intestine

Leishmania (Viannia) braziliensis is one of the main causes of cutaneous leishmaniasis in the Americas. This species presents genetic polymorphism that can cause destructive lesions in oral, nasal, and oropharyngeal tracts. In a previous study, the parasite caused several histopathological changes to hamster ileums. Our study evaluates immune response components, morphological changes, and effects on neurons in the ileums of hamsters infected by three different strains of L. (V.) braziliensis in two infection periods. For the experiment, we separated hamsters into four groups: a control group and three infected groups. Infected hamsters were euthanized 90- or 120-days post infection. We used three strains of L. (V.) braziliensis: the reference MHOM/BR/1975/M2903 and two strains isolated from patients who had different responses to Glucantime® treatment (MHOM/BR/2003/2314 and MHOM/BR/2000/1655). After laparotomy, ileums were collected for histological processing, biochemical analysis, and evaluation of neurons in the myenteric and submucosal plexuses of the enteric nervous system (ENS). The results demonstrated the increase of blood leukocytes after the infection. Optical microscopy analysis showed histopathological changes with inflammatory infiltrates, edemas, ganglionitis, and Leishmania amastigotes in the ileums of infected hamsters. We observed changes in the organ histoarchitecture of infected hamsters when compared to control groups, such as thicker muscular and submucosa layers, deeper and wider crypts, and taller and broader villi. The number of intraepithelial lymphocytes and TGF-β-immunoreactive cells increased in all infected groups when compared to the control groups. Mast cells increased with longer infection periods. The infection also caused remodeling of intestinal collagen and morphometry of myenteric and submucosal plexus neurons; but this effect was dependent on infection duration. Our results show that L. (V.) braziliensis infection caused time-dependent alterations in hamster ileums. This was demonstrated by the reduction of inflammatory cells and the increase of tissue regeneration factors at 120 days of infection. The infected groups demonstrated different profiles in organ histoarchitecture, migration of immune cells, and morphometry of ENS neurons. These findings suggest that the small intestine (or at least the ileum) is a target organ for L. (V.) braziliensis infection, as the infection caused changes that were dependent on duration and strain.

Nano amphotericin B: a good anti-leishmanial drug with effect on cathelicidin gene expression.

Protozoan parasites, such as Leishmania major (L. major), remained as a global health problem of the current century. Leishmania major is a major cause of cutaneous leishmaniasis (CL) in developed and developing countries. Traditionally, amphotericin B is prescribed as an alternative drug, while first-line drugs failed. Some active proteins of the innate immune system such as cathelicidins try to inhibit infection Via several proposed mechanisms. Here this research aimed to not only determine the anti-leishmanial activity of nano amphotericin B but also to evaluate which anti-leishmanial drug can induce the cathelicidin gene expression. Both promastigote and amastigote stages of L. major were exposed to various concentrations of nano amphotericin B, amphotericin B and finally compared to glucan time as standard drug for CL treatment. For the gene expression of cathelicidin, macrophages were exposed to the same concentration of anti-leishmanial drugs. The findings demonstrated that nano amphotericin B was more effective at all concentrations than amphotericin B. Additionally, among tested anti-leishmanial drugs, nano amphotericin B has more potency to induce the cathelicidin gene expression in macrophages cells. The findings revealed that nano amphotericin B has potential as an effective anti-leishmanial drug against CL caused by L. major parasites.

High-resolution habitat suitability model for Phlebotomus pedifer, the vector of cutaneous leishmaniasis in southwestern Ethiopia

BackgroundPhlebotomus pedifer is the vector for Leishmania aethiopica causing cutaneous leishmaniasis (CL) in southwestern Ethiopia. Previous research on the transmission dynamics of CL resulted in recommendations for vector control. In order to target these interventions towards affected areas, a comprehensive understanding of the spatial distribution of P. pedifer at high spatial resolution is required. Therefore, this study determined the environmental predictors that facilitate the distribution of P. pedifer and created a map indicating the areas where conditions are suitable for survival of the vector in southwestern Ethiopia with high spatial resolution.MethodsPhlebotomus pedifer presence points were collected during two entomological surveys. Climate, vegetation and topographic variables were assembled. Climate variables were interpolated with variables derived from high-resolution digital elevation models to generate topoclimatic layers representing the climate conditions in the highlands. A Maximum Entropy model was run with the presence points, predicting variables and background points, which were selected based on a bias file.ResultsPhlebotomus pedifer was the only captured Phlebotomus species in the study area and was collected at altitudes ranging between 1685 and 2892 m. Model projections indicated areas with suitable conditions in a ‘belt’ surrounding the high mountain peaks. Model performance was high, with train and test AUC values being 0.93 and 0.90, respectively. A multivariate environmental similarity surface (MESS) analysis showed that the model projection was only slightly extrapolated for some of the variables. The mean annual temperature was the environmental variable, which contributed most to the model predictions (60.0%) followed by the seasonality in rainfall (13.2%). Variables representing steep slopes showed very low importance to model predictions.ConclusionsOur findings indicate that the suitable habitats for P. pedifer correspond well with the altitudes at which CL was reported previously, but the predictions are more widely distributed, in contrast with the description of CL to occur in particular foci. Moreover, we confirm that vector distribution is driven by climate factors, suggesting inclusion of topoclimate in sand fly distribution models. Overall, our model provides a map with a high spatial resolution that can be used to target sand fly control measures in southwestern Ethiopia.

Genotyping of Cutaneous Leishmaniasis Cases Detected Before and After Migration with Real-Time Polymerase Chain Reaction in Hatay

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. 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. 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. 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.

Molecular Identification of Species Caused Cutaneous Leishmaniasis in Southern Zone of Iran

Leishmania major and Leishmania tropica are two main species causing cutaneous leishmaniasis (CL) in Iran. Recently, Crithidia spp. has also been reported in the wound of patients with CL. In this study, we determined the species causing CL in the southern of Iran and the role of Crithidia spp. in creating skin ulcers. In this cross-sectional study from Apr to Sep 2016, 66 patients with CL referred to Diagnostic Lab of Leishmaniasis, Valfajr Health Center, Shiraz, Iran, were selected. After DNA extraction from the Giemsa stained smears, all samples were amplified in two separate steps using specific primers, firstly, to differentiate Leishmania species and then to identify Crithidia spp. Two species L. major and L. tropica were responsible for 60 and 6 cases, respectively. Moreover, in two patients, mixed infection with Crithidia was confirmed. In mix infection cases, the morphology of the cutaneous ulcers was not different from the wounds of other patients. Leishmania major is responsible for the most common CL in southern Iran. In addition, in two patients with L. major and L. tropica, mix infection with Crithidia was confirmed. The potential role of Crithidia as the main factor for CL and the probability of this parasite to have synergistic effects on Leishmania, as a hypothesis, requires more comprehensive researches on the ambiguity of this protozoon.

Dietary Vitamin D3 Deficiency Increases Resistance to Leishmania (Leishmania) amazonensis Infection in Mice.

The leishmaniases are a group of diseases caused by Leishmania parasites, which have different clinical manifestations. Leishmania (Leishmania) amazonensis is endemic in South America and causes cutaneous leishmaniasis (CL), which can evolve into a diffuse form, characterized by an anergic immune response. Since the leishmaniases mainly affect poor populations, it is important to understand the involvement of immunonutrition, how the immune system is modulated by dietary nutrients and the effect this has on Leishmania infection. Vitamin D3 (VitD) is an immunonutrient obtained from diet or endogenously synthesized, which suppresses Th1 and Th17 responses by favoring T helper (Th) 2 and regulatory T cell (Treg) generation. Based on these findings, this study aims to evaluate dietary VitD influence on L. (L.) amazonensis experimental infection in C57BL/6 and BALB/c mice. Thus, C57BL/6 and BALB/c VitD deficient (VDD) mice were generated through dietary VitD restriction 45 days prior to infection. Both strains of VDD mice showed a more controlled lesion development compared to mice on a regular diet (Ctrl). There were no differences in serum levels of anti-Leishmania IgG1 and IgG2a, but there was a decrease in IgE levels in BALB/c VDD mice. Although CD4+ T cell number was not changed, the CD4+ IFN-y+ T cell population was increased in both absolute number and percentage in C57BL/6 and BALB/c VDD mice compared to Ctrl mice. There was also no difference in IL-4 and IL-17 production, however, there was reduction of IL-10 production in VDD mice. Together, our data indicate that VitD contributes to murine cutaneous leishmaniasis susceptibility and that the Th1 cell population may be related to the resistance of VDD mice to L. (L.) amazonensis infection.