Despite the broad distribution of leishmaniasis among Iranians and animals across the country, little is known about the genetic characteristics of the causative agents. Applying both HSP70 PCR-RFLP and sequence analyses, this study aimed to evaluate the genetic diversity and phylogenetic relationships among Leishmania spp. isolated from Iranian endemic foci and available reference strains. A total of 36 Leishmania isolates from almost all districts across the country were genetically analyzed for the HSP70 gene using both PCR-RFLP and sequence analysis. The original HSP70 gene sequences were aligned along with homologous Leishmania sequences retrieved from NCBI, and subjected to the phylogenetic analysis. Basic parameters of genetic diversity were also estimated. The HSP70 PCR-RFLP presented 3 different electrophoretic patterns, with no further intraspecific variation, corresponding to 3 Leishmania species available in the country, L. tropica, L. major, and L. infantum. Phylogenetic analyses presented 5 major clades, corresponding to 5 species complexes. Iranian lineages, including L. major, L. tropica, and L. infantum, were distributed among 3 complexes L. major, L. tropica, and L. donovani. However, within the L. major and L. donovani species complexes, the HSP70 phylogeny was not able to distinguish clearly between the L. major and L. turanica isolates, and between the L. infantum, L. donovani, and L. chagasi isolates, respectively. Our results indicated that both HSP70 PCR-RFLP and sequence analyses are medically applicable tools for identification of Leishmania species in Iranian patients. However, the reduced genetic diversity of the target gene makes it inevitable that its phylogeny only resolves the major groups, namely, the species complexes.
Animals ; Genetic Variation* ; Humans ; Iran ; Leishmania infantum ; Leishmania major ; Leishmania tropica ; Leishmania* ; Leishmaniasis ; Parasites* ; Phylogeny ; Sequence Analysis*

There is no effective treatment modality available against different forms of leishmaniasis. Therefore, the aim of this study was to improve the penetration and efficacy of selenium and glucantime coupled with niosomes and compared them with their simple forms alone on in vitro susceptibility assays. In this study, the niosomal formulations of selenium and in combination with glucantime were prepared. The size and morphology of the niosomal formulations were characterized and the effectivity of the new formulation was also evaluated using in vitro MTT assay, intra-macrophage model, and gene expression profile. From the results obtained, no cytotoxicity effect was observed for niosomal and simple forms of drugs, as alone or in combination. Niosomal formulations of the drugs significantly showed more inhibitory effects (P≤0.001) than the simple drugs when the selectivity index was considered. The gene expression levels of Interleukin (IL-10) significantly decreased, while the level of IL-12 and metacaspase significantly increased (P≤0.001). The results of the present study showed that selenium plus glucantime niosome possess a potent anti-leishmanial effect and enhanced their lethal activity as evidenced by the in vitro experiments.
Gene Expression ; In Vitro Techniques ; Interleukin-12 ; Interleukins ; Leishmania tropica ; Leishmania ; Leishmaniasis ; Liposomes ; Selenium ; Transcriptome

Cutaneous leishmaniasis (CL) can be seen in 2 forms, zoonotic and anthroponotic, in Iran. In this study, epidemiological aspects of CL were studied during an 8-year period (2009–2016) in city of Kashan, central Iran. The demographic and epidemiological data, including age, sex, occupation, number and site of the lesions, treatment regimen, past history of CL, and season of all patients were gathered from the health centers. Descriptive statistics were used to describe features of the study data. Total 2,676 people with CL were identified. The highest annual incidence was estimated to be 182 per 100,000 population in 2009 and the least was in 2016 (47 per 100,000 population). The highest frequency affected age groups were observed in 20–29 year-old patients (20.9%). More than 51% of the patients were under 30 years old. The maximum frequency of the disease, 1,134 (43.3%), was seen in autumn. The most common location of lesions was hands (61.4%). Most of the patients (81.6%) were treated by systemic glucantime regimen. In the city of Kashan, the incidence rate of the CL disease is significantly higher than many other regions of Iran. To reduce the risk of disease, control of reservoir hosts and vectors of disease, and education of individual protection are strongly recommended.
Education ; Epidemiologic Studies ; Epidemiology ; Hand ; Humans ; Incidence ; Iran ; Leishmania major ; Leishmania tropica ; Leishmaniasis, Cutaneous ; Occupations ; Seasons ; Urbanization

In this study, we carried out extensive in vitro studies on various concentrations of tioxolone along with benzoxonium chloride and their niosomal forms against Leishmania tropica. Niosomes were prepared by the hydration method and were evaluated for morphology, size, release study, and encapsulation efficiency. This study measured leishmanicidal activity against promastigote and amastigote, apoptosis and gene expression levels of free solution and niosomal-encapsulated tioxolone along with benzoxonium chloride. Span/Tween 60 niosome had good physical stability and high encapsulation efficiency (more than 97%). The release profile of the entrapped compound showed that a gradual release rate. The combination of niosomal forms on promastigote and amastigote were more effective than glucantime. Also, the niosomal form of this compound was significantly less toxic than glucantime (P≤0.05). The flowcytometric analysis on niosomal form of drugs showed that higher number of early apoptotic event as the principal mode of action (89.13% in 200 μg/ml). Also, the niosomal compound increased the expression level of IL-12 and metacaspase genes and decreased the expression level of the IL-10 gene, which further confirming the immunomodulatory role as the mechanism of action. We observed the synergistic effects of these 2 drugs that induced the apoptotic pathways and also up regulation of an immunomodulatory role against as the main mode of action. Also, niosomal form of this combination was safe and demonstrated strong anti-leishmaniasis effects highlights further therapeutic approaches against anthroponotic cutaneous leishmaniasis in future planning.
Apoptosis ; Gene Expression ; In Vitro Techniques ; Interleukin-10 ; Interleukin-12 ; Leishmania tropica ; Leishmania ; Leishmaniasis, Cutaneous ; Liposomes ; Methods ; Up-Regulation

Cutaneous leishmaniasis (oriental sore) is usually a self-limited infection of the skin caused by the protozoan Leishmania tropica. The disease is endemic to the Mediterranean, Asia, Africa, and the Middle East. It has been seen in this country among many Korean technical experts and labourers working in the endemic areas of the disease. Our patient had acquired cutaneous leishmaniasis in Saudi Arabia and it had remained active for six months. He had been treated with antimony and metronidazole but failed because of severe side effects. And then we treated the patient witb cryosurgery and the skin lesions were followed by resolution with cosmetically acceptable scar in 4 months. The brief review of literature on the treatment of cutaneous leishmaniasis was undertaken.
Africa ; Antimony ; Asia ; Cicatrix ; Cryosurgery* ; Humans ; Leishmania tropica ; Leishmaniasis, Cutaneous* ; Metronidazole ; Middle East ; Saudi Arabia ; Skin

Cutaneous leishmaniasis (CL) is a protozoan disease which is endemic in Iran. It is transmitted by the Phlebotomus sand fly. The eyelid is rarely involved possibly because the movement of the lids impedes the sand fly from biting the skin in this region. Here, we report 6 rare cases of eyelid CL. The patients were diagnosed by skin scraping, culture, and PCR from the lesions. Skin scraping examination showed Leishmania spp. amastigotes in the cytoplasm of macrophages. Culture examination was positive for Leishmania spp. PCR was positive for Leishmania major and Leishmania tropica. The lesions were disguised as basal cell carcinoma, chalazion, hordeolum, and impetigo. The patients were treated with intramuscular meglumine antimoniate (20 mg/kg/day) for at least 3 weeks. They showed a dramatic response, and the lesions almost completely disappeared. We emphasized the importance of clinical and diagnostic features of lesions, characterized the phylogenetic relationship of isolated parasites, and reviewed the literature on ocular leishmaniasis.
Carcinoma, Basal Cell ; Chalazion ; Cytoplasm ; Eyelids* ; Hordeolum ; Humans ; Impetigo ; Iran ; Leishmania ; Leishmania major ; Leishmania tropica ; Leishmaniasis ; Leishmaniasis, Cutaneous* ; Macrophages ; Meglumine ; Parasites ; Phlebotomus ; Polymerase Chain Reaction ; Psychodidae ; Skin

Leishmania tropica is one of the causative agents of leishmaniasis in humans. Routes of infection have been reported to be an important variable for some species of Leishmania parasites. The role of this variable is not clear for L. tropica infection. The aim of this study was to explore the effects of route of L. tropica infection on the disease outcome and immunologic parameters in BALB/c mice. Two routes were used; subcutaneous in the footpad and intradermal in the ear. Mice were challenged by Leishmani major, after establishment of the L. tropica infection, to evaluate the level of protective immunity. Immune responses were assayed at week 1 and week 4 after challenge. The subcutaneous route in the footpad in comparison to the intradermal route in the ear induced significantly more protective immunity against L. major challenge, including higher delayed-type hypersensitivity responses, more rapid lesion resolution, lower parasite loads, and lower levels of IL-10. Our data showed that the route of infection in BALB/c model of L. tropica infection is an important variable and should be considered in developing an appropriate experimental model for L. tropica infections.
Animals ; Disease Models, Animal ; Female ; Leishmania major/*immunology ; Leishmania tropica/*immunology/*pathogenicity ; Leishmaniasis/*immunology/parasitology/*pathology ; Mice ; Mice, Inbred BALB C ; Treatment Outcome

Leishmania tropica and L. major are etiologic agents of human cutaneous leishmaniasis. Delayed type hypersensitivity (DTH) is an immunologic response that has been frequently used as a correlate for protection against or sensitization to leishmania antigen. In BALB/c mice, L. tropica infection results in non-ulcerating disease, whereas L. major infection results in destructive lesions. In order to clarify the immunologic mechanisms of these 2 different outcomes, we compared the ability of these 2 leishmania species in induction of DTH response in this murine model. BALB/c mice were infected with L. major or L. tropica, and disease evolution and DTH responses were determined. The results show that the primary L. major infection can exacerbate the secondary L. major infection and is associated with DTH response. Higher doses of the primary L. major infection result in more disease exacerbation of the secondary L. major infection as well as higher DTH response. L. tropica infection induces lower DTH responses than L. major. We have previously reported that the primary L. tropica infection induces partial protection against the secondary L. major infection in BALB/c mice. Induction of lower DTH response by L. tropica suggests that the protection induced against L. major by prior L. tropica infection may be due to suppression of DTH response.
Animals ; Disease Models, Animal ; Ear/pathology ; Female ; Foot/pathology ; *Hypersensitivity, Delayed ; Leishmania major/*immunology ; Leishmania tropica/*immunology ; Leishmaniasis, Cutaneous/*immunology/*parasitology/pathology ; Mice ; Mice, Inbred BALB C

Cutaneous leishmaniasis (oriental sore) is usually a self-limited infection of the skin caused by the protozoan Leishmania tropica. The disease is endemic to the Mediterranean, Asia, Africa, and the Middle East. It has been seen in this country among many Korean technical experts and labourers working in the endemic areas of the disease. Our patient had acquired cutaneous leishmaniasis in Saudi Arabia and it had remained active for six months. He had been treated with antimony and metronidazole but failed because of severe side effects. And then we treated the patient witb cryosurgery and the skin lesions were followed by resolution with cosmetically acceptable scar in 4 months. The brief review of literature on the treatment of cutaneous leishmaniasis was undertaken.
Africa ; Antimony ; Asia ; Cicatrix ; Cryosurgery ; Eczema* ; Humans ; Leishmania tropica ; Leishmaniasis, Cutaneous ; Metronidazole ; Middle East ; Saudi Arabia ; Skin

Cutaneous leishmaniasis is a self-limited infection of the skin caused by Leishmania tropica, a protozoan parasite transmitted by Phlebotomus sandfly. This disease is endemic in tropical and subtropical zone, but recently many cases of cutaneous leishmaniasis were reported in Korean among the peoples who had been worked in the Middle East. We experienced five cases of cutaneous leishmaniasis developed in Korean who had been worked in Saudi Arabia and Jordan. Skin lesions of various sized, central ulcerated and marginal elevated, with serosanguinous discharge and some crust were scattered on exposed area, especially both extremities. Histopathologic findings showed hyperkeratosis, acanthosis, pseudoepitheliomatous hyperplasia, dermal infiltration of histiocytes, lymphocytes, plasma cells and neutrophils, and numerous Leishman-Donovan bodies in and around histiocytes. About 1-4 months treatment with trimethoprim-sulfamethoxazole and metronidazole, two cases were healed with hyperpigmented scar and two cases were improved but one case was not responded.
Cicatrix ; Extremities ; Histiocytes ; Hyperplasia ; Jordan ; Leishmania tropica ; Leishmaniasis, Cutaneous* ; Lymphocytes ; Metronidazole ; Middle East ; Neutrophils ; Parasites ; Phlebotomus ; Plasma Cells ; Psychodidae ; Saudi Arabia ; Skin ; Trimethoprim, Sulfamethoxazole Drug Combination ; Ulcer