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      Molecular detection of Leishmania DNA in wild-caught sand flies, Phlebotomus and Sergentomyia spp. in northern Iran

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          Abstract

          Leishmaniasis is currently considered a major health problem in Iran, posing an increasing threat to society's development in various dimensions. This study aimed to detect Leishmania infection in wild-caught sand flies in Sari City, northern Iran. Sand flies were collected using sticky traps, and Leishmania DNA was identified using polymerase chain reaction (PCR) targeting the ITS2-rDNA region, followed by restriction fragment length polymorphism (RFLP) analysis. A total of 138 female sand flies were tested, among which, only 1 specimen of Ph. papatasi (11.11 %) and Ph. major (14.28 %), 4 specimens of Ph. kandelakii (7.27 %) and Se. dentata (8.33 %), and 2 specimens of Se. sintoni (50 %) were naturally infected with L. ( L.) major. This was observed in the ITS2 nested-PCR amplification assays where a ∼ 245 bp PCR band was produced. Also, RFLP analysis by Mnl1 revealed the fragments of 55 and 70 and 120 bp for infected sand flies which are characteristic of L. ( L.) major. Most of the sand flies were unfed, collected during warm season, found indoor. This study reperesents the first molecular detection of L. ( L.) major in wild-caught sand flies, specifically in Ph. papatasi in this region, as well as Ph. kandelakii and Ph. major in Iran and even the world.

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          MEGA11: Molecular Evolutionary Genetics Analysis Version 11

          The Molecular Evolutionary Genetics Analysis (MEGA) software has matured to contain a large collection of methods and tools of computational molecular evolution. Here, we describe new additions that make MEGA a more comprehensive tool for building timetrees of species, pathogens, and gene families using rapid relaxed-clock methods. Methods for estimating divergence times and confidence intervals are implemented to use probability densities for calibration constraints for node-dating and sequence sampling dates for tip-dating analyses. They are supported by new options for tagging sequences with spatiotemporal sampling information, an expanded interactive Node Calibrations Editor , and an extended Tree Explorer to display timetrees. Also added is a Bayesian method for estimating neutral evolutionary probabilities of alleles in a species using multispecies sequence alignments and a machine learning method to test for the autocorrelation of evolutionary rates in phylogenies. The computer memory requirements for the maximum likelihood analysis are reduced significantly through reprogramming, and the graphical user interface has been made more responsive and interactive for very big data sets. These enhancements will improve the user experience, quality of results, and the pace of biological discovery. Natively compiled graphical user interface and command-line versions of MEGA11 are available for Microsoft Windows, Linux, and macOS from www.megasoftware.net .
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            Leishmaniasis: current situation and new perspectives.

            P Desjeux (2004)
            Leishmaniasis represents a complex of diseases with an important clinical and epidemiological diversity. Visceral leishmaniasis (VL) is of higher priority than cutaneous leishmaniasis (CL) as it is a fatal disease in the absence of treatment. Anthroponotic VL foci are of special concern as they are at the origin of frequent and deathly epidemics (e.g. Sudan). Leishmaniasis burden remains important: 88 countries, 350 million people at risk, 500,000 new cases of VL per year, 1-1.5 million for CL and DALYs: 2.4 millions. Most of the burden is concentrated on few countries which allows clear geographic priorities. Leishmaniasis is still an important public health problem due to not only environmental risk factors such as massive migrations, urbanisation, deforestation, new irrigation schemes, but also to individual risk factors: HIV, malnutrition, genetic, etc em leader Leishmaniasis is part of those diseases which still requires improved control tools. Consequently WHO/TDR research for leishmaniasis has been more and more focusing on the development of new tools such as diagnostic tests, drugs and vaccines. The ongoing effort has already produced significant results. The newly available control tools should allow a scaling up of control activities in priority areas. In anthroponotic foci, the feasibility of getting a strong impact on mortality, morbidity and transmission, is high.
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              Phlebotomine Sand Flies (Diptera: Psychodidae) in Iran and their Role on Leishmania Transmission

              Sand fly research has a long history in Iran beginning with the work of Adler, Theodor and Lourie in 1930 and followed by Mesghali’s foundational taxonomic work on sand flies in 1943. Since then, research has been continued unabated throughout the country and official publications report the existence of at least 44 species of sand flies (26 of the genus Phlebotomus and 18 of genus Sergentomyia) in Iran. So far, seven Phlebotomus species and one Sergentomyia species have been collected and described by Iranian researchers for the first time. Natural promastigote infections have been repeatedly found in 13 species of sand flies and modern molecular techniques are used routinely to characterize Leishmania parasite isolates from endemic areas of cutaneous and visceral leishmaniasis. Because of anthropogenic environmental modifications or human population movements, data on phlebotomine sand flies should be regularly updated and verified at least every five years by fieldwork and taxonomy in foci of leishmaniasis, to incriminate vector species of relevance to the ecology of transmission and to support development and implementation of control programs.
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                Author and article information

                Contributors
                Journal
                Parasite Epidemiol Control
                Parasite Epidemiol Control
                Parasite Epidemiology and Control
                Elsevier
                2405-6731
                26 November 2024
                November 2024
                26 November 2024
                : 27
                : e00395
                Affiliations
                [a ]Health Sciences Research Center, Department of Medical Entomology and Vector Control, School of Public Health, Mazandaran University of Medical Sciences, Sari, Iran
                [b ]MSc Student Research Committee, School of Public Health, Mazandaran University of Medical Sciences, Sari, Iran
                [c ]Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran
                [d ]Department of Medical Entomology and Vector Control, School of Public Health, Health Sciences Research Center, Mazandaran University of Medical Sciences, Sari, Iran
                [e ]Iranian National Registry Center for Lophomoniasis and Toxoplasmosis, Toxoplasmosis Research Center, Imam Khomeini Hospital, Mazandaran University of Medical Sciences, Sari, Iran
                [f ]Department of Medical Microbiology and Immunology, School of Medicine, Qom University of Medical Sciences, Qom, Iran
                Author notes
                [* ]Corresponding author. nasibeh.hoseini@ 123456gmail.com
                Article
                S2405-6731(24)00059-X e00395
                10.1016/j.parepi.2024.e00395
                11650325
                39691461
                d3b1c601-c0c7-4f1b-9eb5-17e797a93fca
                © 2024 Published by Elsevier Ltd on behalf of World Federation of Parasitologists.

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 12 May 2024
                : 24 November 2024
                : 25 November 2024
                Categories
                Original Research article

                leishmania (l.) major,ph. kandelakii,ph. major,se. sintoni,se. dentata,ph. papatasi,nested pcr,pcr-rflp,iran

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