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Background: The primary aim of this study is to determine infection to Leishmania parasites in the wild population of Phlebotomus caucasicus and Phlebotomus mongolensis using molecular methods in some important zoonotic cutaneous leishmaniasis foci in Iran. Methods: Sand flies were collected from active colonies of rodent burrows from 16 trapping sites using sticky trap paper. In order to detect and identify of Leishmania parasites in females Ph. caucasicus and Ph. mongolensis, the Nested– PCR amplification of ITS2-rDNA region was performed to generate amplicon with 245bp for Leishmania major, 206bp for L. gerbilli and 141bp for L. turanica. Results: In the current study we found DNA of different gerbil parasites such as L. major and L. turanica, and mixed infection of L. major/L. turanica in Ph. caucasicus and Ph. mongolensis. It should be noted that, in Iran, natural infection with Leishmania parasites is recorded for the first time in this study in Ph. mongolensis. Conclusion: Both species of Ph. caucasicus and Ph. mongolensis not only may participate in the ZCL transmission cycle between reservoir hosts, but also results of this study support the role of these species as secondary vectors in the transmission of leishmaniasis to humans. [ABSTRACT FROM AUTHOR]

BACKGROUND: Updated information on the vectorial capacity of vectors is required in each malarious areas as well in Iran and its neighboring countries such as Afghanistan. The aims of this study were to investigate the potential infection of about 800 specimens collected from malarious areas of Afghanistan and Iran, and to differentiate biological forms of Anopheles stephensi. Method: Two molecular markers, 18S RNA gene subunit and AsteObp1 intron I, were used respectively for investigation Plasmodium infection and identifying the biological forms of An. stephensi. RESULTS: Plasmodium infection was detected in 4 pools of Afghanistan specimens, including An. stephensi, collected from Nangarhar. Individually examination showed infection in 5 An. stephensi (infection rate: 1.25), to P. falciparum (2), P. vivax (2) and a mix infection. Out of five infected specimens, three were intermediate forms and two were mysorensis. No infection was found in specimens collected from Iran (Chabahar County), probably due to the active malaria control program in south-east of Iran. CONCLUSION: The key role of An. stephensi, as a known Asian malaria vector, was re-emphasized in Afghanistan by the results achieved here. The fauna of vectors and the pattern of biological forms of An. stephensi are similar in both countries that urge regional investigations to provide evidence-based and applied data for decision-maker in malaria control. [ABSTRACT FROM AUTHOR]

Background: Mosquito-borne arboviruses such as West Nile (WN), dengue, Rift Valley fever, and Sindbis viruses are reported in Iran, but large-scale studies have not been performed on mosquitoes to find their vectors. A molecular study of the adult mosquitoes (Diptera: Culicidae) for flaviviruses was carried out in a focus of WN infection, Guilan Province, northern Iran. Methods: Mosquito collections were carried out in five stations of two counties (Anzali and Rasht) using light traps, hand catch by manual aspirators and night landing catch during August-September 2013 and 2014. Molecular screening of WN virus and more widely for Flavivirus RNA was carried out using a specific PCR technique. Results: In total, 1015 adult mosquitoes were collected including eight species representing four genera. The most prevalent species were Aedes vexans (33.2%), Culex tritaeniorhynchus (22%), Cx. pipiens (20.7%), and Anopheles maculipennis s.l. (15.6%). Molecular screening was carried out on the 1015 mosquitoes after they were organized as 38 pools according to sex, species and trapping location. None of the pools were positive. Conclusion: Surveillance should be continued while increasing the sampling campaigns due to the presence of wetlands in the region and abundant species which are considered as vectors, feeding on both birds and humans. [ABSTRACT FROM AUTHOR]

Objective: To clarify the epidemiological aspects of visceral leishmaniasis in Kaleybar and Khoda-Afarin districts, north-west of Iran. Methods: A total of 1 420 human (children under 12 years) samples, 101 domestic dogs samples (Canis familiaris), and 577 female sand fly samples were collected. Sera of human and dogs were tested using the direct agglutination test, and sand flies were identified at species level using the microscopic method. Furthermore, a structured questionnaire was applied to evaluate the correlation between the potential risk factors and the related clinical signs/ symptoms with the human and dogs' seropositivity. Results: Totally, 2.18% of human samples were positive at titers≥: 800; among them, 13 cases (41.94%) were above 1:3 200, and clinical symptoms were observed in all of them except for an 11-year old girl. Anti-Leishmania infantum antibodies were found at titers ≥1: 320 in 9.90% of dogs' samples, half of them had at least one sign of canine visceral leishmaniasis. Moreover, 10 Phlebotomus species were identified in the study areas, and Phlebotomus (Larroussius) major group was the predominant species. There are significant correlations between the presence of anti-Leishmania infantum antibodies and the fever (P<0.001), anemia (P=0.001) and weight loss (P=0.016) in children. On the other hand, significant correlations were revealed between the Leishmania infection and the shelter (P=0.039), cutaneous lesion (P=0.005), lymphadenopathy (P=0.001) and weight loss (P<0.001) in the infected dogs. Conclusions: Visceral Leishmania infection is prevalent in rural areas of Kaleybar and Khoda- Afar districts located in East-Azerbaijan province, therefore active detection and treatment of visceral leishmaniasis cases should not be neglected. [ABSTRACT FROM AUTHOR]

Objective: To delineate reliable morphological characteristics for identifying and separating female Phlebotomus caucasicus and Phlebotomus mongolensis which exist sympatrically in the main foci of zoonotic cutaneous leishmaniasis in Iran. Methods: Sand flies were collected using sticky trap papers from active colonies of rodent burrows installed from 16 catching sites. Morphometric measurements were analyzed of 87 Phlebotomus caucasicus and 156 Phlebotomus mongolensis. Univariate and multivariate analysis were carried out to determine significant morphometric variables for discrimination of the two species. Finally, seven morphological characteristics of 65 female Phlebotomus caucasicus and 124 female Phlebotomus mongolensis were described. Results: Univariate and multivariate analyses of 10 morphometric variables via Discriminant Function Analysis (DFA) and Principal Component Analysis (PCA) showed that five morphometric variables had an accuracy of 100% for discriminating female Phlebotomus caucasicus and Phlebotomus mongolensis. Moreover, PCA revealed that the five morphometric variables with the highest loadings separated these two species. Morphological studies on antennal flagellum (and its associated structures) and mouth-parts of female specimens demonstrated significant differences in several structures. Conclusions: The results show that morphological and morphometrical features can be used to discriminate two female isomorphic species, Phlebotomus caucasicus and Phlebotomus mongolensis accurately. [ABSTRACT FROM AUTHOR]

Background: The fatal form of leishmaniasis is visceral form (VL), found in some of the countries in the world. Visceral leishmaniasis has been reported sporadically from all provinces in Iran, including Lorestan. This study aimed to characterize parasite species in DAT positive and some of the DAT negative human blood samples of Delfan district, Lorestan Province, central Iran. Methods: Blood samples were collected from different geographical areas of Delfan. Serum was used for DAT test and remained part of molecular study. DNA was extracted by using DNG-plus extracted kit (Cinagen, Iran). Polymerase chain reaction amplification of Leishmania kDNA and PCR-RFLP of ITS1 was done to identify Leishmania species. Some amplicons were sequenced, submitted to GenBank and analyzed by BLASTn. Results: Expected band of kDNA for L. infantum (720bp) was amplified in 16 out of 186 (8.6%) samples which showed previously anti-Leishmania antibody at different titers or were negative serologically. Using BLASTn, 93% similarity with L. infantum has been shown. The rDNA-ITS1 was amplified only in 9 samples (4.7%). RFLP pattern was similar to what expected for L. infantum. Conclusion: A new emerging hypo-endemic focus, caused by L. infantum, is going to be established in Delphan District, Lorestan Province. Further studies on vector and reservoirs are necessary for the region and other parts of Lorestan Province. [ABSTRACT FROM AUTHOR]

Phlebotomine sandflies are vectors of phleboviruses that cause sandfly fever or meningitis with significant implications for public health. Although several strains of these viruses had been isolated in Iran in the late 1970's, there was no recent data about the present situation at the outset of this study. Entomological investigations performed in 2009 and 2011 in Iran collected 4,770 sandflies from 10 different regions. Based on morphological identification, they were sorted into 315 pools according to species, sex, trapping station and date of capture. A phlebovirus, provisionally named Dashli virus (DASHV), was isolated from one pool of Sergentomyia spp, and subsequently DASHV RNA was detected in a second pool of Phlebotomus papatasi. Genetic and phylogenetic analyses based on complete coding genomic sequences indicated that (i) DASHV is most closely related to the Iranian isolates of Sandfly fever Sicilian virus [SFSV], (ii) there is a common ancestor to DASHV, Sandfly fever Sicilian- (SFS) and SFS-like viruses isolated in Italy, India, Turkey, and Cyprus (lineage I), (iii) DASHV is more distantly related with Corfou and Toros viruses (lineage II) although common ancestry is supported with 100% bootstrap, (iii) lineage I can be subdivided into sublineage Ia including all SFSV, SFCV and SFTV except those isolated in Iran which forms sublineage Ib (DASHV). Accordingly, we suggest to approve Sandfly fever Sicilian virus species consisting of the all aforementioned viruses. Owing that most of these viruses have been identified in human patients with febrile illness, DASHV should be considered as a potential human pathogen in Iran. [ABSTRACT FROM AUTHOR]

Background: Malarious areas in Iran are close to Afghanistan and Pakistan that urge the researchers to extend their knowledge on malaria epidemiology to the neighboring countries as well. Vectorial capacity differs at species or even at population level, the first essential step is accurate identification of vectors. This study aimed to identify Anopheles species composition in selected malarious areas of Afghanistan and Iran, providing further applied data for other research in two countries. Methods: Adults Anopheles spp. were collected from four provinces in Afghanistan (Badakhshan, Herat, Kunduz, Nangarhar) by pyrethrum spray catch, hand collection methods through WHO/EMRO coordination and from Chabahar County in Iran by pyrethrum spray catch method. Identification was performed using reliable identification key. Results: Totally, 800 female Anopheles mosquitos, 400 from each country were identified at species level. Anopheles composition in Afghanistan was An. superpictus, An. stephensi and An. hyrcanus. Most prevalent species in Badakhshan and Kunduz were An. superpictus, whereas An. stephensi and An. hyrcanus were respectively found in Nangarhar and Heart. Anopheles species in Chabahar County of Iran were An. stephensi, An. fluviatilis, An. culicifacies and An. sergentii. The most prevalent species was An. stephensi. Conclusion: Current study provides a basis for future research such as detection of Plasmodium infection in collected samples which is on process by the authors, also for effective implementation of evidence-based malaria vector intervention strategies. [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]

Background: Pediculosis capitis is a common parasitic infection of children, which has been neglected in many areas as well in Shahriar County, Tehran province. This study was conducted to determine the prevalence of pediculosis capitis among pupils in primary schools of Shahriar County. Materials and Methods: This descriptive-analytical study was conducted in Shahriar for the period of 2012 to 2013. Data were collected by questionnaires and direct head inspection. The prevalence rate was determined in three primary schools and 750 girls participated, which were selected by a randomized cluster method. Data were analyzed using Statistical Package for Social Sciences (SPSS16) and Chi-square test. Results: Among the 750 girls who were examined, head pediculosis was observed in 36 cases (4.8%). There was a significant relationship between head lice infestation and two factors, father's job and using private tools in hair salon. Conclusion: The results showed that pediculosis capitis could be a potentially health threatening problem for school children. It is necessary to give health education to families in order to prevent pediculosis in the study areas. [ABSTRACT FROM AUTHOR]

Background: Female sand flies of subgenus Adlerius are considered as probable vectors of visceral leishmaniasis in Iran. The objective of this study was to determine the morphological and genotypic variations in the populations of this subgenus in the country. Methods: Sand flies collected using sticky traps from 17 provinces during 2008-2010. The morphometric measurements were conducted with an Ocular Micrometer. Data was analyzed by SPSS. The Cytb gene was used to estimate population genetic diversity and identify the female specimens. UPGMA phenetic tree was used for DNA haplotypes of Cytb gene. Results: Six species of subgenus Adlerius identified from which one species, P. (Adlerius) kabulensis, is new record. The identification key is provided for males. Results revealed the molecular systematic in the species of subgenus Adlerius and determine the relationship of three females of P. comatus, P. balcanicus and P. halepensis. Conclusion: The positions of three females and the males in the UPGMA tree are correct and the similarities among them confirm our results. The branches of each species are not genetically distinct which justify the overlapping morphological characters among them. Molecular sequencing of Cytb-mtDNA haplotypes can be used for female identification for different species of subgenus Adlerius in Iran. [ABSTRACT FROM AUTHOR]

Background and Objectives: Leishmaniasis is one of the most important diseases in Iran, with high prevalence in some part of country including, Tehran province. This study was aimed to investigate the cutaneous lesions of patients studies was also referred to different laboratory of Health center of Varamin (a city in Tehran province), sand flies species in selected area. Material and Methods: In this cross sectional study demographic data was collected by special questionnaire. Smears of suspected patients to stained and examined by microscope. Sand flies were collected by sticky traps, CDC light traps and aspirators, female specimens were mounted and identified by diagnostic keys at species level. Results: Leishmania parasites were observed in 56.82% of cases. Disease was more common among male (64%). The majority of patients (30%) aged between 25-39 year old. Most lesions (50%) observed in hands and legs of patients, 76% of affected people had just one lesion. Half of them had a history of travel to endemic areas. More than 2500 sand flies captured during this study; among them 1100 female specimens were mounted and identified. The prevalent spicies was Sergentomyia spp (65/9%), the second was. papatasi (33/20%). It should be mentioned that Ph. sergenti, Ph. alexandri and Ph. caucasicus group also were identified in lower percentage. Conclusion: According to current results and the status of the studied region including it seems that (Agricultural activities, animal husbandry and migration), cutaneous leishmaniasis could be a health threatening problem. [ABSTRACT FROM AUTHOR]

Background and Objectives: Two clinical forms of leishmaniasis exist in Iran: cutaneous and visceral. According to the sporadic reports of new cases of Visceral Leishmaniasis (VL) in Lorestan province, real status of VL is not clear, so this study aimed to describe the seroprevalence of VL in Delphan city. Materials and Methods: In this descriptive analytic study, blood samples were collected from children ≤ 12 years and 10% of adults by a multi-stage randomized cluster sampling from January 2012 to September 2012. The sero-prevalence evaluation was done by Direct Agglutination Test (DAT). Based on different studies in Iran, the 1/800 and 1/1600 titers were considered as the infection with L. infantum and the of titers ≥ 1/3200 accompanied with clinical symptoms was considered as VL disease. Results: 800 collected serum samples, 21(2.62%) showed anti-Leishmania antibodies at titers of 1/800 and 1/1600, whereas 5(0.62%) showed anti-Leishmania antibodies at titers of ≥1/3200. But just one of them showed clinical symptoms (anemia and large abdominal) which is under treatment with miltefosine. Conclusion: A new focus of VL with low endemicity is going to be formed in our region, which showed that further studies on vector and reservoirs is necessary in the region and other parts of Lorestan province. [ABSTRACT FROM AUTHOR]

Background: Stigmaeids mites have been recorded only on Phlebotominae sand flies up to now. Five species of Eustigmaeus, and three of Stigmaeus were reported on infested sandflies in different country up to the present. Methods: Sand flies collection was done using CDC light trap and sticky paper. The mites were isolated from infested specimens, mounted in Puri's medium and identified using reliable keys. Results: A mite infested Phlebotomus papatasi was observed during a study on sandflies of one of the southern provinces of Iran, near to the Persian Gulf. Several scars resulting from mite attachment were found on abdominal tergites of this female sand fly. The mites were identified as Eustigmaeus johnstoni. Conclusion: This parasitic mite is one of the eyeless species, which has a great distribution over the world, reported from Tunisia, Saudi Arabia, Pakistan, Yemen, Cyprus and Palestine. But, this is the first record of this species from Iran. [ABSTRACT FROM AUTHOR]

Abstract: An intraspecific study on the morphological and molecular characteristics of Phlebotomus sergenti s.l., the main vector of Leishmania tropica, was performed on 28 Iranian populations from 11 provinces and a few samples from Greece, Morocco, Lebanon, Turkey, Pakistan, and Syria. Three morphotypes were identified as A, B and C, with some intermediate forms in the samples under investigation. Based on the number of setae and the width of basal lobe of coxite, differences between A and B morphotypes were highly significant. Excluding one unusual haplotype, sequence analysis of ∼439bp of mtDNA (a fragment of cytochromeB gene, tRNA for serine gene, and a fragment of NADH1 gene) revealed a 6–7% genetic distance within the Iranian populations and among the specimens of other countries. Neighbor–Joining (NJ) analysis confirmed the existence of three main groups within our samples. Although there was no consistency between morphotypes and genotypes, but an interrelationship was found between morphometry and morphotypes. Morphotype A, which was considered as P. sergenti sergenti, was the most prevalent in collection sites. Morphotype B, which was identified as Phlebotomus sergenti similis, is the first record of this subspecies in Iran, and was found to be sympatric with other morphotypes. Morphotype C had an elongated style in comparison with P. sergenti sergenti. Molecular database showed three main genetic structures. This is the first combined morphological and molecular studies on P. sergenti s.l. in Iran. [Copyright &y& Elsevier]

Background and Objectives: Phlebotominae sand flies are small, fragile, nocturnal Nematoceran insects with weak flight capability; they transmit viral and bacterial disease, including leishmaniasis. Sand fly-parasite relationship, vector behavior, taxonomy and its ecology studies depend on sampling methods. In this study the efficiency of different trapping method has been evaluated for the first time in Bushehr province that is an endemic foci of Cutaneous leishmaniasis in Iran. Materials and Methods: Collection of sand flies was done by different methods. During each village visit, sand flies were collected by CDC light traps, sticky traps and aspirator from indoors dwellings and for outdoors, CDC, UV light traps and sticky traps were applied. Results: Phlebotomus papatasi was the most common species captured both in outdoor and indoor traps. Comparing the collection methods, maximum samples were captured by CDC light traps (40.5%), followed by aspirator and then sticky trap in indoors environment. Whereas most effective method at outdoor was UV trap (47.6%), followed by sticky trap and CDC light trap. Conclusion: Based on biodiversity indices, UV light trap captures wider of species. It seems that light trap is more efficient and less labor-intensive but more work is needed to evaluate the efficiency another trap such as frozen CO2 and other attractants. [ABSTRACT FROM AUTHOR]

Leishmaniasis is a protozoal and vector-borne disease. World health organization has considered the disease as a neglected tropical disease. Phlebotomus and Lutzumyia species (order: Diptera, family: Psychodidae) are human leishmaniasis vectors in new and old worlds. Sergentomyia spp. (Diptera, Psychodidae) are proven vectors of lizard leishmaniasis. Although some studies have identified human Leishmania parasites in Sergentomyia , their role in parasite circulation is unknown yet. Hence, the parasitological and molecular methods were used to study the possible  Leishmania  infection of  Sergentomyia  spp., in the human and canine visceral leishmaniasis endemic area in North West of Iran. Even though Sergentomyia specimens were caught in a dominant number compared to Phlebotomus spp., no Leishmania promastigote or DNA was detected in live-caught or sticky trap-caught specimens, respectively. Sergentomyia spp. are proven vectors of sauroleishmaniasis, and despite several global reports of Leishmania infection in Sergentomyia spp., such findings should be carefully interpreted to avoid false vector incriminations., Competing Interests: There is no conflict of interest in this study., (© 2023 Urmia University. All rights reserved.)

Background: Leishmaniasis is a zoonotic disease caused by species of protozoa of the genus Leishmania . In recent years, incidence of cutaneous leishmaniasis has increasing trend in Golestan Province, North of Iran. The aim of the present study was to identify the frequency of cutaneous leishmaniasis using PCR-RFLP in patients referred to Kalaleh Health Center, during 2013-14., Methods: This descriptive cross-sectional study was conducted on 70 individuals with suspected cutaneous leishmaniasis that referred to health center of Kalaleh County, Golestan Province, Northern Iran, from Sep 2013 to Nov 2014. Samples of cutaneous lesions were examined microscopically. DNA was extracted from all of the positive smears and PCR was done on ITS-1 gene. RFLP was performed using HaeIII enzyme for species identification., Results: Totally, 38 out of the 70 (54.3%) suspected individuals including 22 males (57.9%) were found positive by microscopic examination. All of microscopically positive samples were confirmed to be positive for Leishmania DNA (approximately 340 bp bands were detected). RFLP revealed 140 bp and 200 bp bands (approximate size), indicative of L. major ., Conclusion: The detected species of studied region was L. major . Cutaneous leishmaniasis has high prevalence in Kalaleh County, thus more studies on leishmaniasis in the animal reservoirs, comparison of homology of animal and human isolates and a survey regarding natural infection of vectors in this region is highly recommended.

Background: Female sand flies of subgenus Adlerius are considered as probable vectors of visceral leishmaniasis in Iran. The objective of this study was to determine the morphological and genotypic variations in the populations of this subgenus in the country., Methods: Sand flies collected using sticky traps from 17 provinces during 2008-2010. The morphometric measurements were conducted with an Ocular Micrometer. Data was analyzed by SPSS. The Cytb gene was used to estimate population genetic diversity and identify the female specimens. UPGMA phenetic tree was used for DNA haplotypes of Cytb gene., Results: Six species of subgenus Adlerius identified from which one species, P. (Adlerius) kabulensis, is new record. The identification key is provided for males. Results revealed the molecular systematic in the species of subgenus Adlerius and determine the relationship of three females of P. comatus, P. balcanicus and P. halepensis., Conclusion: The positions of three females and the males in the UPGMA tree are correct and the similarities among them confirm our results. The branches of each species are not genetically distinct which justify the overlapping morphological characters among them. Molecular sequencing of Cytb-mtDNA haplotypes can be used for female identification for different species of subgenus Adlerius in Iran.