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Background: Cutaneous Leishmaniasis (CL) is a parasitic disease with diverse outcomes. Clinical diversity is influenced by various factors such as Leishmania species and host genetic background. The role of Leishmania RNA virus (LRV), as an endosymbiont, is suggested to not only affect the pathogenesis of Leishmania, but also impact host immune responses. This study aimed to investigate the influence of LRV2 on the expression of a number of virulence factors (VFs) of Leishmania and pro-inflammatory biomarkers. Materials and methods: Sample were obtained from CL patients from Golestan province. Leishmania species were identified by PCR (LIN 4, 17), and the presence of LRV2 was checked using the semi-nested PCR (RdRp gene). Human monocyte cell line (THP-1) was treated with three isolates of L. major with LRV2 and one isolate of L. major without LRV2. The treatments with four isolates were administered for the time points: zero, 12, 24, 36, and 48 h after co-infection. The expression levels of Leishmania VFs genes including GP63, HSP83, and MPI, as well as pro-inflammatory biomarkers genes including NLRP3, IL18, and IL1β, were measured using quantitative real-time PCR. Results: The expression of GP63, HSP83, and MPI revealed up-regulation in LRV2 + isolates compared to LRV2- isolates. The expression of the pro-inflammatory biomarkers including NLRP3, IL1β, and IL18 genes in LRV2- were higher than LRV2 + isolates. Conclusion: This finding suggests that LRV2 + may have a probable effect on the Leishmania VFs and pro-inflammatory biomarkers in the human macrophage model. [ABSTRACT FROM AUTHOR]

Background: Recent studies have shown an increasing number of patients with cutaneous leishmaniasis (CL) who do not respond to pentavalent antimonials as the first line of treatment for CL. Nanocarriers such as extracellular vesicles (EVs) are efficient vehicles that might be used as drug delivery systems for the treatment of diseases. Therefore, we aimed to isolate and characterize the EVs of Leishmania major, load them with Amphotericin B (AmB), and investigate the toxicity and efficacy of the prepared drug form. Methods: The EVs of L. major were isolated, characterized, and loaded with amphotericin B (AmB), and the EVs-Amphotericin B (EVs-AmB) form was synthesized. Relevant in vitro and in vivo methods were performed to evaluate the toxicity and efficacy of EVs-AmB compared to the control. Results: The anti-leishmanial activity of the EVs-AmB showed a higher percentage inhibition (PI%) (P = 0.023) compared to the AmB at different concentrations and time points. Obtained data showed a significant increase in the lesion size and parasite load in the lesion, PBS, and EVs mice groups in comparison with EVs-AmB, AmB, and Glucantime groups (P < 0.05), EVs-AmB had a significant decrease in lesion sizes in comparison with AmB (P < 0.05). Results showed that EVs-AmB decreased its toxicity to the kidneys and liver (P < 0.05). Conclusion: EVs-AmB improved the efficacy of AmB in mouse skin lesions and reduced hepatorenal toxicity. Furthermore, EVs could be a promising nanoplatform for the delivery of AmB in CL caused by L. major. [ABSTRACT FROM AUTHOR]

Purpose: Cutaneous leishmaniasis (CL) is a major vector-borne disease that affects people globally, including Iran. Different factors are associated with leishmaniasis pathogenicity; recently, a link of the possible relationship between Leishmania RNA Virus (LRV) and disease severity was proposed, especially in the New World leishmaniasis (NWL). This study was aimed to investigate the presence of LRV2 in Leishmania isolates in Aran o Bidgol, Isfahan province. Methods: Samples were collected from 110 CL-suspected patients referred to the health center. In this study, we aimed to investigate CL cases (parasitologically and clinically), identify Leishmania species (by ITS1-PCR–RFLP), and finally detection of LRV2 (by RdRp-semi-nested PCR). Results: Parasitological methods showed 60 positive cases, based on the HaeIII enzyme restriction profile, 59 cases were caused by L. major and 1 case by L. tropica. Our project is the first study on LRV2 isolation in Aran o Bidgol city and the LRV was successfully detected from a single L. major isolated in a women's hand lesion. Using BLAST, 94.8–100% similarity was observed in the RdRp sequence of current LRV isolate with those available in GenBank from Iran or overseas. Conclusion: L. major was the main cause of CL in Aran o Bidgol, although L. tropica is also present in a much lower proportion in the area. This is the first report on the presence of LRV2 in Aran o Bidgol and the fifth in Iran. [ABSTRACT FROM AUTHOR]

Background: Cutaneous leishmaniasis (CL) is one of the world health problems. Leishmania major is the etiological agent of zoonotic cutaneous leishmaniasis (ZCL). Promastigote and amastigote are two morphological forms of Leishmania parasites that express different proteins and p27 is an important gene encoding cytochrome c oxidase (COX) component. P27 gene expresses a 27 kDa protein that essential in ATP synthesis. This study aimed to compare p27 gene expression in promastigote and amastigote forms in Iranian strain of L. major (MRHO/IR/75/ER). Methods: This study was conducted in 2015. Clinical isolates of CL patients from north, center, west and south parts of Iran were collected and identified by PCRRFLP. After RNA extraction of promastigotes and amastigotes and cDNA synthesis, the expression level of p27 gene was compared by real-time RT-PCR. Results: By comparison of expression level between amastigote and promastigote forms of Iranian strain of L. major, up-regulation of p27 gene (2.73 fold) was observed in amastigotes. Moreover, there was no significant difference in p27 gene expression between L. major isolates. Conclusion: p27 gene and protein can be considered as a target in recombinant vaccine production and treatment process. [ABSTRACT FROM AUTHOR]

Background: Cutaneous leishmaniasis (CL) is one of the most important neglected tropical diseases and a major public health challenge in Iran caused by Leishmania spp and transmitted by phlebotomine sand flies. The number of CL cases has shown an increasing pattern all over the country, including the district of Varamin, southeast of Tehran, Iran. This study aimed to identify the Leishmania spp isolated from CL patients using molecular methods in Varamin during 2012-2013. Methods: Exudate materials collected from the swollen edge of the skin lesions of 44 parasitological positive CL patients by disposable lancet. They were referred to Varamin Health Center by physician. The samples were subjected to molecular method for Leishmania species identification. Results: The digestion pattern of restriction enzyme revealed that 37 (84.1%) CL patients were infected with L. major and 7 (15.9%) were infected with L. tropica. They were mostly male than female. More than half of the patients (58%) had multiple lesions, and they were mostly observed on extremities, 34.1% on legs and 29.5% on hands. Le- sions were mostly of wet ulcerative type. Conclusion: Dominancy of L. major provides more evidence that Varamin District probably could be considered as Zoonotic Cutaneous Leishmaniasis (ZCL) areas. More investigation on other epidemiological aspects of disease is needed. [ABSTRACT FROM AUTHOR]

Introduction: Cutaneous leishmaniasis (CL) is among the top 10 infectious disease priorities in the world, and the leading cause of morbidity in Iran. The present study was conducted to assess the risk of CL, and to determine some epidemiological features of the disease in endemic areas of Qom Province in Central Iran during 2009 to 2013. Methods: Data regarding human cases of the disease were obtained from the Qom Province Health Center, prepared and stored in a spatial database created in ArcGIS10.3. A total of 9 out of 212 Leishmania spp. positive slides taken in 2013 from patients residing in Qom city were examined using molecular methods and the species of Leishmania was identified by PCR-RFLP. Those 9 patients had no history of travel outside the city. Spatial analysis and clustering methods were applied to find major hot spots and susceptible areas for the establishment of novel foci of the disease. Transmission patterns were examined for spatial autocorrelation using the Moran's I statistical application, and for the clustering of high or low values using the Getis-Ord Gi* statistics. Results: During the period of study, a total of 1767 CL cases were passively reported in the area, out of which were 65% males and 35% females. The highest and lowest numbers of cases were reported in 2010 and 2013, respectively. Importantly, 979 cases were reported from urban areas, while the remainder came from rural areas. Leishmania major was detected as the causative agent of CL in the city of Qom. Remarkably, most patients recorded in Qom city were associated with a history of travel to the endemic areas of CL within the province, or to other endemic areas of the disease in Iran. Spatial distribution of CL cases revealed northeastern and southwestern quarters of the city were the major hot spots of the disease (P<0.05). Hot spot and CL transmission risk analysis across the province indicated that more than 40 villages were located in high and very high risk areas of CL transmission. Conclusions: Due to the existence of hot spots (P<0.05) of CL in successive years in some quarters of Qom city, along with detection of L. major from the patients without a history of travel, there may be potential of local transmission of the disease within the city. Therefore, it is necessary to conduct a comprehensive study concerning the hot spots of CL in Qom city for curtailing the incidence of the disease in the city. The methodology and the results of this study is essential in serving as a yardstick for subsequent similar studies that will be carried out in other endemic areas of CL in Iran and providing an adequate tool for the establishment of a national database of cutaneous leishmaniasis. [ABSTRACT FROM AUTHOR]

Background In the northeast and central parts of Iran, Rhombomys opimus (great gerbil) is the primary reservoir host of zoonotic cutaneous leishmaniasis (ZCL). This study used both parasitological and molecular methods to identify Leishmania spp. and their different haplotypes that were circulating in the great gerbil populations in ZCL foci from northeastern Iran. Methods A cross-sectional study using microscopy, culturing and molecular methods was conducted to detect Leishmania parasites in 194 live R. opimus in ZCL foci from northeastern Iran during 2010–2011. Results Leishmania spp. were found in 38.1% (74/194) of the samples by microscopy and in 41.2% (80/194) by culturing. Small papules and skin thickening on the upper edge of the ears were observed in 25 (12. 9%) of the R. opimus. PCR-RFLP and PCR direct sequencing of internal transcribed spacer 1 (ITS1) rRNA showed similar infection rates for L. major and L. turanica in 60 eligible R. opimus, only one mixed infection containing both L. major and L. turanica was found. Phylogenetic analysis was conducted using the ITS1 sequences of 32 isolates that were successfully aligned by comparison of their base sequences with the ITS1 DNA sequence database using ClustalW and MEGA5. The samples were classified into monophyletic clusters (>97% bootstrap). Six haplotypes were observed for L. major and seven for L. turanica. Conclusion In northeast Iran, L. major, and L. turanica naturally circulate in R. opimus, and L. major/L. turanica co-infections also exist. Phylogenetic analysis suggested that Leishmania spp. isolated from R. opimus are not a monophyletic group. [ABSTRACT FROM AUTHOR]

• Among the 181 gerbils captured, 48.6% were Leishmania spp. positive upon microscopic examination. • Leishmania infection was observed in 89.5% of the gerbils using nested-PCR. • Single or mixed infections of L. major with L. gerbilli or L. turanica were detected in 28.4% gerbils. • Homology investigation using NCBI blast against Genbank database sequences revealed 92.45–100% homology between samples and L. major in the Genbank sequences using ITS2-rDNA. Rodents from the subfamily Gerbillinae are the principal reservoir of cutaneous leishmaniasis (CL) caused by Leishmania major in the center and northeast of Iran. This study was conducted to determine both naturally occurring Leishmania infection rates and the distribution of Leishmania species in the central parts of Iran during 2019–2020. In this regard, presence of Leishmania parasites were confirmed by microscopic examination and the species were identified by nested-PCR using the Internal Transcribed Spacer2- Ribosomal DNA (ITS2-rDNA). Finally, some samples were sequenced and, the blast analysis of L. major samples, showed a 92.45–100% homology to the L. major sequence. Of the 181 wild gerbils collected (Rhombomys opimus =157 and Meriones lybicus =24), 88 (48.6%) tested positive for Leishmania sp. by microscopic examination whereas 162 (89.5%) were positive by nested-PCR. Of the 162 infected gerbils, 103 showed single strain infections (30 L. major, 28 L. gerbilli and 45 L. turanica) , 43 showed dual infections with only the non-human species (L. gerbilli and L. turanica) , and 16 were mixed infections of L. major and L. turanica (n = 14) or L. gerbilli (n = 2). All single or mixed L. major infections were detected in gerbils from areas with reports of human CL during the last decade. These findings suggest that Rhombomys opimus and Meriones libycus have a potential role in the maintenance of human and non-human transmission of Leishmania species in the CL foci. [ABSTRACT FROM AUTHOR]