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Background Leishmaniasis is a major parasitic disease worldwide, except in Australia and Antarctica, and it poses a significant public health problem. Due to the absence of safe and effective vaccines and drugs, researchers have begun an extensive search for new drugs. The aim of the current study was to investigate the in vitro leishmanicidal activity of larval saliva and hemolymph of Lucilia sericata on Leishmania tropica. Methods The effects of different concentrations of larval products on promastigotes and intracellular amastigotes of L. tropica were investigated using the mouse cell line J774A.1 and peritoneal macrophages as host cells. The 3-(4.5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and direct observation and counting method were used to assess the inhibitory effects and cell cytotoxicity of the larval products. The effects of larval products on the amastigote form of L. tropica were quantitatively estimated by calculating the rate of macrophage infection, number of amastigotes per infected macrophage cell, parasite load and survival index. Results The 50% cytotoxicity concentration (CC50) value of both larval saliva and hemolymph was 750 µg/ml, and the 50% inhibitory concentration (IC50) values were 134 µg/ml and 60 µg/ml for larval saliva and larval hemolymph, respectively. The IC50 for Glucantime, used a positive control, was (11.65 µg/ml). Statistically significant differences in viability percentages of promastigotes were observed for different doses of both larval saliva and hemolymph when compared with the negative control (p ≤ 0.0001). Microscopic evaluation of the amastigote forms revealed that treatment with 150 µg/ml larval hemolymph and 450 µg/ml larval saliva significantly decreased the rate of macrophage infection and the number of amastigotes per infected macrophage cell. Conclusion Larval saliva and hemolymph of L. sericata have acceptable leishmanicidal properties against L. tropica. Graphical Abstract

Background: The attraction of phlebotomine sand flies to plant and animal hosts is due to the produced chemical compounds, affecting the olfactory receptors of the insects. Therefore, novel and effective methods, such as Attractive Toxic Sugar Baits (ATSB) and Attractive Toxic Baits (ATB), are based on the effective materials that attract sand flies toward the host. The present study was designed to identify the attractive materials in plants and animals for using in ATSB and ATB. Methods: This cross-sectional study was carried out in July 2018 on endemic areas of leishmaniasis in Iran. Different baits, including mango, nectarine, grape, banana, melon and watermelon, defibrinated blood of cattle, sheep, goat and chicken, urine of cattle, sheep, goat and ultimately, simple and complex chemicals, such as CO2, 1-octanol, lactic acid and human sweat were placed inside the traps, and the rate of the sand flies attraction to these materials was studied. Furthermore, data were analyzed using the Kruskal-Wallis test and Mann Whitney U test. Results: There was a significant difference in the sand flies attraction between the traps containing watermelon, urine of cattle, and sheep, and chemicals such as CO2 and human sweat and the control trap (p< 0.05). Conclusion: This study showed that watermelon and CO2 are the potential candidates for using in ATSB and ATB, respectively.

Salivary proteins specific antibodies have been shown to be useful biomarkers of exposure to sand fly bites. This study aimed to investigate the level, duration, and dynamics of the human immune response against the SGL of Phlebotomus sergenti Parrot, 1917 (Diptera: Psychodidae), and to assess the immunoreactivity of human sera with SGL components in an endemic area of anthroponotic cutaneous leishmaniasis (ACL) in Iran. The study was carried out in 2-phase; longitudinal and cross-sectional. Sand flies were collected monthly from indoors and outdoors. In the longitudinal study, sera from healthy volunteers were collected monthly, and in the cross-sectional study, sera from healthy volunteers and patients with ACL lesion/s, were collected for immunoassay studies. The level of anti-P. sergenti saliva IgG was detected using the ELISA. Immunoreactivity of individual human sera with saliva components was also assessed by western blotting. Phlebotomus sergenti was the predominant sand fly species in the study area. The maximum and minimum percentages of IgG responses were seen in October (66%) and March (29%), respectively. Additionally, the cross-sectional study showed that 59.3% of the healthy volunteers and 80% of the patients were IgG positive. The antibody response against P. sergenti salivary gland was high during the sand fly active season and declined by the end of the activity of the vectors. Antibody response against the SGL components of P. sergenti was transient and individual-specific. Some individuals shared a strong reaction against certain individual antigens, which could be considered as vector exposure markers for further investigation. LIST OF ABBREVIATIONS: ELISA: Enzyme-Linked Immunosorbent Assay; SDS PAGE: Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis; SGL: Salivary Gland Lysate; ACL: Anthroponotic Cutaneous Leishmaniasis; PBS: Phosphate Buffered Saline; BCA: Bicinchoninic Acid; PBS-T: Phosphate Buffered Saline Tween; FBS: Fetal Bovine Serum; HRP: Horseradish Peroxidase; TMB: 3,3′,5,5′-Tetramethylbenzidine; PVDF: Polyvinylidene Difluoride; SGA: Salivary Gland Antigens; OD: Optical Density; KDa: Kilodalton; VL: Visceral Leishmaniasis; CL: Cutaneous Leishmaniasis; SGs: Salivary glands

Background: Visceral and cutaneous leishmaniasis are common in some areas of Iran and consider as health problems. Phlebotomus alexandri has been incriminated as a suspected vector for the both form of leishmaniasis. Methods: This study was carried out in 4 western provinces of Iran. Sand flies were collected using sticky traps and light traps from indoor and outdoor resting places. Nested PCR was employed to detect Leishmania parasites among collected sand flies. Results: Seven hundred and twenty two P. alexandri females were collected and pooled in 179 batches. Results of nested PCR showed, out of 9 samples from East Azerbaijan Province, only one sample was infected by Leishmania infantum. Of 34 individual and pooled samples from Kermanshah Province, only one pooled sample was infected with Leishmania major and among 30 individual and pooled samples in Fars Province, five specimens were infected by L. major, L. infantum, Leishmania donovani and Leishmania tropica. Furthermore, out of 108 individual and pooled samples from Khuzestan Province, 10 samples showed infection with L. major and L. infantum. Conclusion: The results of this study showed that P. alexandri is more active in hot zones than in moderate zones and this species may be considered as a permissive species.

Background & objectives: Phlebotomine sandflies (Diptera: Psychodidae) transmit several important zoonotic diseases to humans and leishmaniasis is one of them. Two types of leishmaniasis, viz. visceral leishmaniasis (VL) and cutaneous leishmaniasis (CL) are endemic in Iran. The main vector of anthroponotic cutaneous leishmaniasis (ACL) is Phlebotomus sergenti. The aim of the present study was to determine the susceptibility status of wild strain of P. sergenti to different imagicides of DDT, bendiocarb and permethrin at the median lethal time, LT50 level. Methods: Sandflies were collected from selected village in North Khorasan Province, northeast of Iran from indoors using CDC light-traps. Susceptibility test was carried out against DDT (4%), bendiocarb (0.1%) and permethrin (0.75%) for all the females according to WHO method, and mortality was calculated. Species identification was carried out using the morphological keys. Data were analysed using probit regression analysis to determine the LT50 and LT90 values. Results: In total, 851 female P. sergenti sandflies were tested. LT50 values to DDT (4%), Bendiocarb (0.1%) and permethrin (0.75%) were 15.4, 19.2 and 6.3 min respectively. The values for LT90 were 51.1, 47.4 and 18.6 min respectively. The mortality rates for 1 h exposure time to DDT, bendiocarb and permethrin were 89.8 ± 1.4; 93.6 ± 1.4; and 95.6 ± 1.7%, respectively. Interpretation & conclusion: The suscesptibility studies revealed development of resistance against DDT (4%) in the wild strain of P. sergenti population. Monitoring and mapping of insecticide resistance in the region is recommended for vector control.

Background: Leishmaniasis represents the important public health problem in the world. One of the main objectives of World Health organization is capacity building of managers and authorities who are involved with diseases control ac­tivities. Methods: The course was conducted in Esfahan Health Research and Training Center (E.H.R.T.C) in summer 2005 and 2009. The course carried out jointly by the Ministry of Health and Medical Education (MOH) of Iran, World Health Or­ganization-Eastern Mediterranean Regional Office (WHO-EMRO) and School of Public health, Tehran University of Med­ical Sciences (SPH-TUMS) and designed for medical officers, senior technicians and managers involved in leish­mania­sis control. Prior to initiate the course, pre-test evaluations including different subjects on leishmaniasis and its control were carried out. The examinations include multiple choice questions. The duration of the course was 3 weeks. A total of 206 contact hours were taught. The main subjects were Basic epidemiology, Leishmaniasis parasitology, Leishmaniasis en­tomology, control of vectors and reservoirs, principles of integrated pest management, Field work and Planning. Dif­ferent methods of teaching including lecture, laboratory, workshop, team work, field exercise and presen­tation were used. Requirement for achievement of the course was to have at least 60% of the total mark for awarding the diploma certificate. Results: A total of 45 participants from Iraq, Afghanistan and Iran graduated from this course. Conclusion: The course is providing the skill for managers, how to combat against disease in their country and is paral­lel to the policy of the leishmaniasis control for capacity building in endemic areas of their countries.

Use of sterile fly larvae (maggots) of blow flies for the treatment of many different types of skin and soft tissue wounds is called Maggot debridement therapy (MDT). The larvae of blow flies secrete a broad spectrum of compounds with diverse mechanisms of action in the gut and salivary glands called excretion/secretion (ES) products which showed to have antimicrobial activities against Gram negative and positive bacteria. Cutaneous leishmaniasis (CL) which is the common form of leishmaniasis is difficult to treat. In this study, the effect of ES from 2nd and 3rd stages of L. sericata and C. vicina larvae on in vitro Leishmania major amastigote growth in macrophage was evaluated. The effect of ES on Leishmania growth was estimated by assessing the rate of macrophage infection and the number of amastigotes per infected macrophages. In addition, the anti Leishmania activities of larval and ES of L. sericata and C. vicina on the skin lesion induced by L. major infection was evaluated in susceptible BALB/c mice. The results showed that ES of both flies reduced the number of infected macrophages; 2.6 and 1.5-fold using L. sericata ES and C. vicina ES, respectively, and inhibited amastigotes growth in macrophages; 2.03 and 1.36-fold by L. sericata ES and C. vicina ES, respectively as compared to the control group. The results showed that L. sericata ES was significantly more effective than C. vicina ES to inhibit in vitro L. major amastigotes growth, The size of lesion was significantly smaller in BALB/c mice treated with L. sericata ES than treated with C. vicina ES. The results of in vivo experiments suggested that pre-treatment with ES derived from L. sericata may have some protective effects on the development of L. major lesion. Therefore, it seems that maggot ES might be considered as a possible candidate for the treatment of cutaneous leishmaniasis.

Background & objectives: Cutaneous leishmaniasis (CL) is still a public health threat in many rural areas of Iran. It causes considerable morbidity among a huge number of people in the endemic areas. Khuzestan is considered as a CL focus in Iran and recently several cases have been reported from Shush County. The aim of the current study was to determine different aspects of CL epidemiology in this County. Methods: The study was conducted in four villages in the rural district of Shush County. In order to survey the scar and acute rate of the disease, 100 inhabitants were questioned in each village. In addition, to determine the reservoir host and vector of the disease, rodents were captured using Sherman traps, and sandflies were collected from around the rodent burrows using sticky paper traps. Results: The scar and acute lesion rates were reported as 5.26 and 0.57% among inhabitants, respectively. Fifty-four rodents including Tatera indica (55.5%), Nesokia indica (33.3%), Mus musculus (9.3%) and Rattus norvegicus (1.9%) were captured and identified. Nested PCR showed 3 out of 54 rodents infected with Leishmania major. In total, 1122 sandflies were collected, and Leishmania major detected in Phlebotomus papatasi (4.35%), Sergentomyia dentata (23%), and Sergentomyia clydei (10%). Interpretation & conclusion: Zoonotic cutaneous leishmaniasis due to L. major prevailed in this area. The common and the main vector was Ph. papatasi and, T. indica seems to be the main while N. indica as a secondary reservoir host of the disease in this area.

Background Cutaneous leishmaniasis (CL) remains a serious public health concern in some parts of Iraq. The aims of this study to report of CL in some parts of Iraq, by different parasitological, cultural, and molecular methods and evaluate sex differences among infected patients. This is the first study conducted to characterize Leishmania species causing CL among Iraqi patients using the sequence analysis of Internal Transcribed Spacer1(ITS1) at Wasit Province. Methods A total of 700 cases of suspected CL were referred to the Iraqi clinics and health centres and they checked for Leishmania amastigote using a Giemsa-stain; however, the Novy Macneal Nicolle (NNN) culture led to the growth of promastigotes in all samples, then the samples were examined using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP)-PCR methods. Results The present study indicated that the prevalence of CL as follow: AL-Diwaniyah 88(15.1%), Wasit 85 (14.5%), Najaf 79 (13.6%), Thi-Qar76 (13.1%), Basrah 67 (11.5%), Baghdad 65 (11.2%), Diyala 63(10.8%), and Salah-Edin province which recorded lower infection 60 (10.3%), and it appeared in 83.3% by using Giemsa- smeared stain. The highest infection rate (100%) appeared using PCR while the lowest infection (68%) appeared by culture on NNN media. The present study was revealed that the highest infection (60%) was caused by L.major rather than L.tropica (40%). Our results showed that 368 (52.6%) of CL patients were had single lesion and 215 (30.7%) had multiple lesions, and the ulcerative wet type lesions were present in 49.6%, while the nodule dry type lesions were present in 33.7%. The overall prevalence of cutaneous leishmaniasis in the study area was very high (83.3%) having a statistically significant association with sex; males are more prone (56.4%) to CL as compared to females (43.6%) Conclusions A clear and reliable bias toward males exists in some tropical diseases, such as leishmaniasis. CL is a major health problem in Iraq and CL caused by many countries including Iraq. Health authorities should be aware of the fact that war and terrorist activities induce expansion of the disease and increase the incidence rate in the situation that access to medical treatment is not easy especially in poor conditions in leishmaniasis endemic areas.

Visceral leishmaniasis (VL) is an important vector-borne disease in Iran. Till now, Leishmania infantum has been detected from five species of sand flies in the country including Phlebotomus kandelakii, Phlebotomus major s.l., Phlebotomus perfiliewi, Phlebotomus alexandri and Phlebotomus tobbi. Also, Phlebotomus keshishiani was found to be infected with Leishmania parasites. This study aimed at predicting the probable niches and distribution of vectors of visceral leishmaniasis in Iran. Data on spatial distribution studies of sand flies were obtained from Iranian database on sand flies. Sample points were included in data from faunistic studies on sand flies conducted during 1995-2013. MaxEnt software was used to predict the appropriate ecological niches for given species, using climatic and topographical data. Distribution maps were prepared and classified in ArcGIS to find main ecological niches of the vectors and hot spots for VL transmission in Iran. Phlebotomus kandelakii, Ph. major s.l. and Ph. alexandri seem to have played a more important role in VL transmission in Iran, so this study focuses on them. Representations of MaxEnt model for probability of distribution of the studied sand flies showed high contribution of climatological and topographical variables to predict the potential distribution of three vector species. Isothermality was found to be an environmental variable with the highest gain when used in isolation for Ph. kandelakii and Ph. major s.l., while for Ph. alexandri, the most effective variable was precipitation of the coldest quarter. The results of this study present the first prediction on distribution of sand fly vectors of VL in Iran. The predicted distributions were matched with the disease-endemic areas in the country, while it was found that there were some unaffected areas with the potential transmission. More comprehensive studies are recommended on the ecology and vector competence of VL vectors in the country.

A molecular study was carried out to incriminate sand fly vectors of cutaneous leishmaniasis (CL) in rural areas of Sarakhs district, Khorassane-Razavi Province, northeastern Iran, in 2011. Sand flies of Sergentomyia with three species and Phlebotomus with six species respectively comprised 73.3% and 26.7% of the specimens. Phlebotomus papatasi was the most common Phlebotomine species in outdoor and indoor resting places. Leishmania infection was found at least in 17 (22%) specimens including Ph. papatasi (n=9 pool samples), Phlebotomus caucasicus (n=6), Phlebotomus alexandri (n=1), and Sergentomyia sintoni (n=1). The parasites were found comprised Leishmania major (n=5), Leishmania turanica (n=10), and Leishmania gerbilli (n=4). Infection of Ph. papatasi with both L. major and L. turanica supporting the new suggestion indicating that it is not restricted only with L. major. Circulation of L. major by Ph. alexandri, and both L. gerbilli and L. turanica by Ph. caucasicus, in addition to previous data indicating the ability of Ph. alexandri to circulate Leishmania infantum and Leishmania donovani, and Ph. caucasicus to circulate L. major, suggests that these two species can be permissive vectors. The results suggest that Ph. papatasi and Ph. alexandri are the primary and secondary vectors of CL where circulating L. major between human and reservoirs, whereas Ph. caucasicus is circulating L. turanica and L. gerbilli between the rodents in the region.

Leishmaniasis is a Neglected Tropical Disease (NTD) and emerging parasitic infection that affect mainly poor regions around the world. This study aimed to determine the baseline susceptibility of Phlebotomus papatasi to commonly used insecticides in a hyper endemic area using WHO standard procedure in central Iran. A total of 4-5 replicates containing 120-200 sand flies were used for each insecticide. Baseline susceptibility to DDT and pyrethroids was assessed on 5326 specimens collected from the study area. The LT50 and LT90 values were measured according to the World Health Organisation test using probit analysis and regression lines. The test results against males P. papatasi revealed that LT50 values to DDT 4%,Permethrin 0.75%, Deltamethrin 0.05%, Cyfluthrin 0.15% and Lambdacyhalothrin 0.05% were 564.07, 38.08, 1.95, 0.60 and 9.78s and the figures for females were 584.44, 110.10, 11.64, 1.53 and 16.91s, respectively. Our results indicated that P. papatasi as the main cutaneous leishmaniasis vector was susceptible to Cyfluthrin 0.15%, Lambdacyhalothrin 0.05%, Permethrin 0.75% and Deltamethrin 0.05% and tolerant to DDT 4%. This study was carried out in one out of many Leishmaniasis foci in Iran. We recommend that future studies incorporate other regions and use the same procedure for monitoring and evaluating sand fly resistance. Also, WHO can provide a specific guideline and create a test kit for sand fly resistance monitoring and for applying susceptibility test because the tubes prepared for mosquitoes are not actually fit for sand flies.

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

Zoonotic cutaneous leishmaniasis (ZCL) is transmitted to humans by phlebotomine sand fly bites. ZCL is a major health problem in Iran, where basic knowledge gaps about sand fly species diversity persist in some ZCL-endemic areas. This paper describes the richness and spatial distribution of sand fly species, collected with sticky traps, in Qom province, a ZCL-endemic area in central Iran, where sand fly fauna has been poorly studied. Collected species were mapped on urban and rural digital maps based on a scale of 1/50,000. All analyses were undertaken with rural- and urban-level precision, i.e., rural and urban levels were our basic units of analysis. After identifying the sand flies, high-risk foci were determined. For spatial analysis of vector species population, the entomological sampling sites were geo-referenced using GPS. Arc GIS 9.3 software was used to determine the foci with leishmaniasis vector species. Following the analyses, two genera (Phlebotomus and Sergentomyia) and 14 species were identified. Based on the mapping and sand fly dispersion analysis, the rural districts were categorized into three groups-infection reported, without infection, and no report. Based on Geographical Information System analyses, Kahak and Markazi districts were identified as high-risk foci with leishmaniasis vector species. These findings can act as a help guide to direct active control measures to the identified high-risk foci and, eventually, lead to reduction in incidence of the disease.

Accurate identification of animal reservoirs of transmissible diseases is an absolute requirement to any epidemiological survey of zoonoses and is essential for predicting species-specific population outbreaks and therefore to develop accurate ecological control strategies. The systematic status of the great gerbil (Rhombomys opimus) remains unclear, despite the fundamental role of these rodents as the main known reservoir hosts of the protozoan parasite Leishmania major in the epidemiology of zoonotic cutaneous leishmaniasis (ZCL) in central and south Asia. In the present work, we represent molecular evidence supporting the identification of at least two major lineages (subspecies) within the species of R. opimus in Iran. Analysis of the mitochondrial cytochrome b (cyt b) gene, revealed a range of 1–10% genetic variation among populations, which were well associated with biogeographic origins and subspecies. Results of laboratory cross hybridization between the subspecies and finding sympatric haplotypes of the two subspecies suggested that no pre- or post-zygotic barriers exist between the subspecies indicating that they still belong to a single taxon. However, the amount of genetic variations between populations/subspecies is high enough to lead them to speciation in future. Implications of such findings on the eco-epidemiology of ZCL in Iran are discussed.

Many rodent species act as reservoir hosts of zoonotic cutaneous leishmaniasis in endemic areas. In the present study a simple and reliable assay based on nested PCR was developed for the detection and identification of Leishmania parasites from rodent skin samples. We designed Leishmania-specific primers that successfully amplified ITS regions of Leishmania major, Leishmania gerbilli and Leishmania turanica using nested PCR. Out of 95 field collected Rhombomys opimus, 21 were positive by microscopic examination and 48 by nested PCR. The percentage of gerbils infected with L. major, L. gerbilli and L. turanica was 3.2%, 1.1% and 27.4%, respectively. In 15.8% of the rodents, we found mixed natural infections by L. major and L. turanica, 1.1% by L. major and L. gerbilli, and 2.1% by the three species. We concluded that this method is simple and reliable for detecting and identifying Leishmania species circulating in rodent populations.

Objective To reveal subspecies composition of Rhombomy opimus ( R. opimus ) in Iran. Methods In this study, field specimens of the gerbil were collected from all its geographical rang in northeast and central parts of Iran and identified on basis of morphological characteristics. Results Results revealed presence of two subspecies of R. opimus sodalis and R. opimus sargadensis in the country. The first subspecies with brown to hazel-nut color and a size bigger than a typical R. opimus was found only in Golestan province, an isolated colony which is located in plains of north slopes of Alborz Chain Mountains of the country, connected with the Turkmenistan. The subspecies of R. opimus sargadensis with a yellowish color and similar size as a typical R. opimus was found in all other areas of the great gerbil distribution in Iran. There was a distinct topographic difference but similar ectoparasites between colonies of the two subspecies in the study area. Conclusions Further ecological and genetic investigations are required for more detailed description of the R. opimus subspecies range and structure. The R. opimus – Phlebotomus papatasi-Leishmania major association and the ZCL severity and outcome in hosts are discussed.

Cutaneous Leishmaniasis (CL) is the most important health problem in the city of Bushehr, southwestern Iran. The objective of the study was to determine some ecological aspects of sand flies in the city during 2010-2011. Sand flies were collected monthly from outdoors and indoors by sticky traps at four selected districts of the city. They were also dissected and examined by Nested-PCR for identification of the parasite during August- September of 2011. A total of 1234 adult sand flies were collected and 6 species including 3 of Genus Phlebotomus and 3 of Genus Sergentomyia were identified. Four species including P. papatasi (3.98 %), P. sergenti (1.14%), S. tiberiadis (87.18%) and S. baghdadis (7.7%) were found indoors. Six species including P. papatasi (3.47%), P. sergenti (3.17%), P. alexandri (0.1%), S. tiberiadis (77.74 %), S. baghdadis (15.41 %) and one female of S. clydei (0.11%) were collected from outdoors. Sand flies started to appear from March and disappear at the end of January. There was only one peak in the density curve in July. The study revealed that S. tiberiadis and S. baghdadis could enter indoors which 89 % and 81.8 % of them were found blood-fed respectively. Moreover, P. papatasi, S. tiberiadis and S. baghdadis were active indoors and outdoors in most months of the year. Nested-PCR of P. papatasi females was positive against kinetoplast DNA of L. major and L. turanica and also mixed natural infections were found by L. gerbilli and L. turanica. Moreover mixed infections by L. major and L. turanica were observed in this species. Sergentomyia clydei and S. tiberiadis were found to be negative to any DNA of Leishmania species. Phlebotomus sergenti females were found infected with DNA of L. turanica and this is the first report of natural infection and detection of the parasite from this sand fly species in worldwide.

An outbreak of cutaneous leishmaniasis [CL] in Sabzevar county prompted this study of the epidemiology and the ecology of vectors and reservoirs. Examination of 541 schoolchildren showed rates of 9.4% for scars and 5.9% for ulcers. Among 807 inhabitants of 4 villages, 10.4% had scars and 3.0% had active lesions. The most highly infected age group was 0-4 years with a rate of 5.9%. A total of 12 849 sandflies representing 7 species were collected in the study area. Leptomonad infection was found in Phlebotomus papatasi, P. caucasicus and Sergentomyia sintoni. Parasites from man, P. papatasi and Rhombomys opimus, were isolated and characterized as Leishmania major. Based on this survey, this is an epidemic of zoonotic CL, with R. opimus the main reservoir host, and P. papatasi the main vector

Introduction Attractive Toxic Sugar Baits (ATSB) is a new vector control method that meets Integrated Vector Management (IVM) goals. In an experimental design, this study aimed to determine effects of ATSB on control of Phlebotomus papatasi, as a main vector of Zoonotic Cutaneous Leishmaniasis (ZCL), in Qom Province, center of Iran. Methods In a cross-sectional design, boric acid was mixed with brown sugar solution and tested as toxic baits for P. papatasi. Two methods were utilized to use the baits: (a) spraying ATSB on vegetation, bushes, and shrubs; and (b) setting ATSB-treated barrier fences in front of colonies at 500 m distance from the houses in outskirts of villages. In order to examine the residual efficacy rate of ATSB-treated barrier fences, the bioassay test was used. Density of P. papatasi sandflies was measured using sticky and light traps biweekly. For data analysis, Mann-Whitney U Test and Kruskal-Wallis were used. Results ATSB-treated barrier fences led to 3 times reduction in P. papatasi population. Besides that, ATSB spraying on plants led to more than 5 times reduction in P. papatasi population. Conclusions Comparing the incidence of leishmaniasis in treated villages before and after the study showed that the incidence was statistically reduced. Therefore, ATSB is an effective method to control vectors and prevent leishmaniasis.

Leishmaniasis is a major vector-borne disease and health problem in Iran. Studies on sand flies, as the vectors of the disease, began in the Northern and Western parts of the country in 1930 and have been continued up to now. Concerning many published information in the field of sand flies, providing a digital database for the country will help the public health authorities to make more correct and prompt decisions for planning leishmaniasis control programs as well as modeling and forecasting of transmission potential across the country. All published data on phlebotomine sand flies of Iran were collected. A database was then designed in Excel format, including all available information regarding sand flies. The valid data were transferred to ArcGIS9.3 to prepare the first spatial database of sand flies of Iran. The IrSandflybase includes 131 papers, 2 abstracts and 71 PhD/MSc theses, reporting studies conducted during 1930–2012. This database contains different available data covering all aspects of ecology and biology of 50 sand fly species in two genera of Phlebotomus and Sergentomyia in the country. The temporal activity of sand flies is reported 9 months in warm regions of the southern part, while it may reduce to 7–8 months in central plateau or 4–5 months in cold areas of the northwest. Occasional studies reported rare species from the borderlines of Iran. It seems that changing the climate due to global warming may affect the spatial distribution of different species and expand it into the country, the issue that can be followed by an updated database.

We report four disseminated cutaneous leishmaniasis(DCL) cases referred to leishmaniasis laboratory at the School of Public Health, Tehran University of Medical Sciences with multiple nodular, ulcerative and crusted lesions extended on the face, trunk, and extremities. None of the patients had any complication and historical involvement in their immunological system conditions that suggest as the criteria for DCL. Direct smears of ulcers were positive for Leishmania parasite. The parasite was isolated from the active lesions and identified as Leishmania major (L. major) using PCR-RFLP assay and sequencing analysis.

Zoonotic cutaneous leishmaniasis (ZCL) due to Leishmania major is a great public health problem in the Old World. Leishmania major is widely distributed in populations of rodents in arid and savannah regions. In this study, seasonal variation of natural infection with Leishmania parasites in Rhombomys opimus (Rodentia: Gerbillinae) population of an endemic focus of ZCL in Iran was monitored. The study was conducted from October 2007 to October 2008 in the central part of the country. Nested polymerase chain reaction (PCR) assay was used for the detection and identification of Leishmania parasites, and the results were confirmed by PCR-restriction fragment length polymorphism (RFLP). The results showed that Leishmania infection rate was 55.8% (29 out of 52 gerbils) using nested PCR. The highest and lowest Leishmania infection rates were observed in fall and summer, respectively. Gerbils that were found to be infected only with L. major were 5.8%, and that with Leishmania turanica were 23.1%. A mixed natural infection was seen in the rodents with L. major and L. turanica (21.2%), with L. major and L. gerbilli (1.9%), and with all the three species (3.9%). Leishmania major infection alone was seen in fall and winter whereas mixed infection of L. major and L. turanica was observed in all seasons except in summer. Leishmania turanica infection was observed throughout the year. It is concluded that L. major, L. gerbilli, and L. turanica circulate in the population of R. opimus in central part of Iran. Leishmania major infection is usually accompanied by L. turanica in naturally infected gerbils with the highest rate in fall. It is recommended that the role of L. turanica in the epidemiology and transmission of ZCL should be reconsidered.

Background: Anthroponotic cutaneous leishmaniasis (ACL) has long been a significant public health prob-lem in northeastern Iran. The objective of this study was to determine the efficacy of deltamethrin-impregnated vs. nonimpregnated bed nets (NIBs) and curtains (NICs) in ACL control. Patients: Deltamethrin-impregnated bed nets (IBs) and curtains (ICs) with 25 mg ai/m2 were distributed among 160 households in one district and NIBs and NICs were distributed among the same number of households in another district. A third district with a similar numbers of households served as a control. Health education mes-sages were disseminated to ensure the population′s complicance with the proper use of bed nets and curtains. Sticky paper traps were used to assess the effect of insecticide-impregnated bed nets and curtains on the density of Phlebotomus sergenti. Deltamethrin susceptibility and also bioassay tests were carried out on the species by WHO standard method. Case findings were done by house-to-house visits once a season and all the inhabitants of the selected households in each district were examined. Results: IBs and ICs provided good protection against sandfly bites and reduced the transmission of ACL in the intervention district, while NIBs and NICs provided no protection. There was no significant difference in monthly density of P. sergenti indoors and outdoors among the districts (P>0.05). This species was susceptible to delta-methrin in the field population in the area. Bioassays confirmed that the nets treated with deltamethrin remained effective for more than 3 months. Conclusion: Personal protection is an effective and sustainable means of preventing and controlling ACL and can reduce dependence on insecticides. We encourage the use of IBs and ICs to control ACL in other high-risk areas of Iran and Afghanistan during the active season of sandflies.

Background: Due to the resurgence of zoonotic cutaneous leishmaniosis (ZCL) in some nonendemic areas of Iran, extensive studies have been conducted on the epidemiology of the disease in different parts of the country in recent years. The objective of this study was to determine the effects of rodent control on the reduction of the incidence of ZCL in an endemic area in Badrood, Iran. Patients and Methods: A survey was carried out in a circle around two villages between April 1997 and January 1999. The control strategy adopted in 1997 consisted of the destruction of the colonies of gerbilline rodents by digging in a radius of 500 m from houses in the intervention area. Opened burrows were baited with zinc phosphide. One village was used as control. Evaluation was made in 1998 and no other control measures were carried out in the area. Case findings were done by house-to-house visits once every season during 1997 and 1998, and all the inhabitants of the selected villages were examined. Results: The average reduction of rodent holes was calculated at 87.4% one year after the first baiting in the intervention area. Changes in the number of rodent holes and the incidence of the disease in the intervention and control villages were statistically significant ( P-value

BACKGROUND : Zoonotic cutaneous leishmaniasis (ZCL) is a serious and increasing public health problem in many rural areas of Iran. Previous studies showed that rodent control operations are effective in destroying rodents and reducing the incidence of ZCL on a small scale and in special circumstances. The objective of this study was to determine suitable timing for rodent control operations in reducing the incidence of ZCL in an endemic area in Badrood, Iran. METHODS : We conducted rodent control operations by baiting rodent holes using zinc phosphide once a month in May, June, July and September in 1997 within a 500-meter circle of houses in the intervention area. From 1999 to 2002, the numbers of active rodent holes were counted in May and October in the intervention and control areas. When the numbers of rodent holes increased to 30% or more of the number before the first baiting in May 1997, holes in the intervention area were baited again with zinc phosphide in the intervention area; no baiting was done in the control village. Case findings were done by house-to-house visits once every season during 2000 to 2002. RESULTS : Changes in the number of rodent holes over time in the intervention and control villages were statistically significant (P< 0.000001). There were also significant differences in the incidence of ZCL between the intervention and control villages (P< 0.005) during 2000 to 2002. The incidence of ZCL was unchanged in the years 2000 and 2001, but increased in one village in 2002 after an increase in the number of rodent holes. CONCLUSION : We suggest that rodent control operations using zinc phosphide be done within a 500-meter circle of houses once every two years before the beginning of the active season of sandflies, which begins in late April.

An endemic focus of zoonotic cutaneous leishmaniosis caused by Leishmania major zymodeme MON-26 was identified in Ardestan town central Iran, during 1998-99. Among 1960 school children examined over a 1-year period of time, 0.92% had evidence of active lesions and 1.53% had scar indicative of past infection. The incidence of the disease among school children was 3.2 per thousand in 1998. In a separate study, the incidence of infection among 100 households in an area with a population of 460 persons showed 3.26% for scars and 1.3% for ulcers. The most highly infected age group was 10-14 with a rate of 2.74%. Meriones libycus (93.3%) and Nesokia indica (6.7%) were present around the town of Ardestan. Both were found to be infected with Leishmania. Two out of 28 M. libycus (7.1%) and one out of two N. indica had positive results. M. libycus is probably the principal reservoir host of zoonotic cutaneous leishmaniosis in this area. Fourteen dogs appeared to be uninfected because examination showed no active lesion or scar. Most probably Phlebotomus papatasi is the vector because about 93% of indoor sandflies were of this species. The active season of sandflies was from late May to late October. Economic changes in this area of Iran may have contributed to this outbreak of cutaneous leishmaniosis.

A total of 22 982 sandflies representing six species were collected using sticky (oiled) paper traps in Badrood rural district, Natanz county from May 1995 to May 1996. Phlebotomus papatasi was the predominant species. The average indoor resting density of P. papatasi ranged from 11.8 (May 1995) to 135.8 per 10 traps (September 1995). The average indoor resting density of this species varied between 39.7-701.3 per 10 traps in May and September, respectively in 1995. The activity of P. papatasi starts from late April or early May and extends to mid October with two peaks, one in mid or late June and the second one in early or mid September. Natural leptomonad infection was found in P. papatasi and Sergentomyia sintoni. P. papatasi could be a vector of humans and gerbils and it is susceptible to DDT in this area. (C) 1999 Elsevier Science B.V. All rights reserved.

In the study carried out in the rural district of Borkhar (Isfahan, Iran) from March 2003 to March 2004 efficacy of deltamethrin impregnated bed nets (IBs) and curtains (ICs), polyester mesh size 156 holes/ inch2, (25 holes/cm2) were compared to that of non-impregnated bed nets (NIBs) and curtains (NICs), in relation to zoonotic cutaneous leishmaniasis (ZCL) control. Deltamethrin impregnated bed nets and curtains at 25 mg a.i./m2 were distributed among 168 households in one of the villages and NIBs plus NICs among the same number of households in another village. In the third village similar numbers of households were used as control. Health education messages were disseminated to ensure the population's compliance with the proper use of bed nets and curtains in the two intervention areas. Entomological surveys using sticky paper traps were carried out in ten households in each village for the collection of sand flies from indoors and outdoors, every two weeks to assess the impact of insecticide impregnated bed nets and curtains on the density of Phlebotomus papatasi. Deltamethrin susceptibility tests and also bioassay tests were carried out on this species by WHO standard method. Case findings were done by house-to-house visits once a season and all the inhabitants of the selected households in each village were examined. The results showed that IBs and ICs can provide a definite personal protection against sand fly bites and interrupted the transmission of ZCL in the intervention village. NIBs and NICs did not provide any protection against the disease. There was no significant difference in monthly density of P. papatasi in indoors and outdoors among the villages (P > 0.05). Susceptibility tests showed that this species was susceptible to deltamethrin in the field population in the area. Bioassays confirmed that the treated nets with deltamethrin remain effective for more than three months and was an excellent mean of individual protection. It is recommended that IBs and ICs can be used widely in the control of ZCL in other similar foci such as hyperendemic and mesoendemic areas of Iran.