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Background and Objective: Cutaneous leishmaniases with two forms of rural and urban is the endemic diseases and as a health problem in our country. Identification of parasite species and type of disease is very important for treatment of disease as well as for planning of control program. The microscopic observations by Giemsa-stained smears is the most common laboratory test for the diagnosis of cutaneous leishmaniasis, but the determination of parasite species is impossible and utilization of other ways such as biochemical and molecular methods is required. This study was carried out to determine the parasite species caused cutaneous Leishmaniasis by Nested PCR in Damghan, Iran. Materials and Methods: This descriptive study was performed on 67 patients with dermal lesions that referred to Damghan health center laboratory in Iran during 2008. The patient's information were recorded in questionnaire. DNA of Giemsa-stained slides from patients was extracted and evaluated by specific primers of kinetoplast DNA using Nested PCR. Results: Leishmania parasites were observed in 57 patients under light microscope. The 10 patients were infected by other dermal diseases. The PCR result showed the parasite presence in lesions of 57 patients is Leismania major. 54% of patients were male and 46% were female. 72% of the patients were lived in rural areas. 50.9% of disease was observed in over 25 years old patients. Hands were the most common region of ulcer (44.7%). 48% of the patients had one ulcer and the other patients had two or more ulcers. High prevalence (31.6%) of disease was observed in October. Conclusion: This study showed that zoonotic cutaneous leishmaniasis to be prevalent in this area and Nested PCR method is a sensitive and accurate to leishmania species characterization.

An Interventional field trial study was carried out in the Dehdahst area, Kohgiloyeh & Buyer Ahmad province, Iran, during 1997-1998. Efficacy of deltamethrin-impregnated bednet at target dosage of 25 a.i. mg/m2 and the incidence of malaria were compared in treated and untreated villages. In this area Anopheles superpictus is the dominant species and the activity of the species occurs in one peak, in August. Regarding to malaria incidence, the results showed a high reduction of Annual Parasite Incidence from 51.2 to zero in treated villages. Bioassay tests on impregnated bednet showed 100% mortality after 110 days and decreased to 97% after 120 days. Due to the exophilic and exophagic habit of vectors in treated areas, it could be concluded that the use of deltamethrin impregnated bed net could reduce the malaria incidence.

Background: Phylogenetic analyses of closely related species of mosquitoes are important for better understanding the evolution of traits contributing to transmission of vector-borne diseases. Six out of 41 dominant malaria vectors of the genus Anopheles in the world belong to the Maculipennis Group, which is subdivided into two Nearctic subgroups (Freeborni and Quadrimaculatus) and one Palearctic (Maculipennis) subgroup. Although previous studies considered the Nearctic subgroups as ancestral, details about their relationship with the Palearctic subgroup, and their migration times and routes from North America to Eurasia remain controversial. The Palearctic species An. beklemishevi is currently included in the Nearctic Quadrimaculatus subgroup adding to the uncertainties in mosquito systematics., Results: To reconstruct historic relationships in the Maculipennis Group, we conducted a phylogenomic analysis of 11 Palearctic and 2 Nearctic species based on sequences of 1271 orthologous genes. The analysis indicated that the Palearctic species An. beklemishevi clusters together with other Eurasian species and represents a basal lineage among them. Also, An. beklemishevi is related more closely to An. freeborni, which inhabits the Western United States, rather than to An. quadrimaculatus, a species from the Eastern United States. The time-calibrated tree suggests a migration of mosquitoes in the Maculipennis Group from North America to Eurasia about 20-25 million years ago through the Bering Land Bridge. A Hybridcheck analysis demonstrated highly significant signatures of introgression events between allopatric species An. labranchiae and An. beklemishevi. The analysis also identified ancestral introgression events between An. sacharovi and its Nearctic relative An. freeborni despite their current geographic isolation. The reconstructed phylogeny suggests that vector competence and the ability to enter complete diapause during winter evolved independently in different lineages of the Maculipennis Group., Conclusions: Our phylogenomic analyses reveal migration routes and adaptive radiation timing of Holarctic malaria vectors and strongly support the inclusion of An. beklemishevi into the Maculipennis Subgroup. Detailed knowledge of the evolutionary history of the Maculipennis Subgroup provides a framework for examining the genomic changes related to ecological adaptation and susceptibility to human pathogens. These genomic variations may inform researchers about similar changes in the future providing insights into the patterns of disease transmission in Eurasia., (© 2023. The Author(s).)

Malaria is the most important mosquito-borne disease, which is transmitted by Anopheles species. According to the recent report of World Health Organization, there were an estimated 229 million malaria cases in 2019 in 87 malaria endemic countries. There are several vector control method specially using pesticides. The aim of this study was to collect all the data about insecticide resistant of malaria vectors in the Eastern Mediterranean region countries. In this study, all the published papers related to insecticide resistance of malaria vectors in the region were collected and resistant status of vectors in the countries was mapped. Resistance to different insecticide classes such as pyrethroids, organophosphate, organochlorine and carbamates were evaluated using World Health Organization guidelines. Results showed a wide variety of susceptibility/resistance status to these chemicals according to the location, historical context of pesticide used, genetic background of vectors, age and abdominal conditions of adults may play a role in the susceptibility status of these species to different insecticides. The results of this study is providing a guideline for countries to manage their vector control activities against insecticide resistance of malaria vectors and provide novel approaches such as male sterility, using Wolbachia, using new insecticide with new mode of action., (© The Author(s) 2022. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Background: We aimed to evaluate the rate of infection by endoparasites amongst rodents in the western regions of Iran to enhance the level of knowledge amongst health authorities in this entity., Methods: This study was conducted in the west and southwest of Kurdistan Province, including the cities of Sanandaj, Marivan, and Sarvabad. The field mission of this work was performed in three seasons' spring, summer, and autumn. The rodents were captured alive and their gastrointestinal tracts were evaluated for the worm endoparasites., Results: Herein, 208 rodents from 15 types of seven species, including Apodemus , Meriones , Mus , Sciurus , Cricetulus , Microtus , and Dryomys , were captured. In addition, 67 (32.21%) rodents were infected with endoparasites and 10 types of worms endoparasites, including Syphacia muris , Streptophagus spp., Mastophorus muris , Skrjabinema spp., Trichostrongylus spp., Trichuris muris , Hymenolepis nana, Hymenolepis dimimuta , Heligmosomoide spp ., and other oxiuros were isolated from their gastrointestinal tracts. Most of these parasites (60%) were isolated from their small intestine whereas they were least (10%) found in their stomach and cecum., Conclusion: Having compared the results of this study with other studies in different regions of Iran, there is a higher variety of rodents and worm parasites in these regions of Iran., Competing Interests: Conflict of interest The authors declare that there is no conflict of interest., (Copyright © 2022 Mohammadi et al. Published by Tehran University of Medical Sciences.)

Malaria and leishmaniasis are the public health problems in southern Iran. The main activity of vector control is indoor residual spraying using pyrethroids, using different insecticides as larviciding and impregnated bednets. The aim of study was to evaluate the biological assays of bendiocarb wettable powder (WP) at different surfaces of wall. The residual effect of bendiocarb WP80 at 400 milligram/meter square (mg/m 2 ) was evaluated on various local surfaces of rooms such as mud and plaster as well as thatch roofs and wooden. World Health Organization standard cones using contact bioassays were carried out using laboratory reared sugar-fed, 48-72 h old females of Anopheles stephensi . Contact bioassays were carried out on sprayed surfaces for 150 days. Contact bioassay on surfaces treated with bendiocarb WP80 at different surfaces was estimated about 2 months. Fumigant tests of bendiocarb WP80 at 400 mg/m 2 revealed 50-93.83% mortality with 1 month persistency. The results showed that carbamate insecticide could be used as rotation with pyrethroids for malaria vector control. Monitoring and evaluation of environmental toxicology of pesticides is important for decision making for choosing appropriate pesticides for disease vector control., (© The Author(s) 2021. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Background: Malaria resurgence has occurred in the northern half parts of Iran. The resurgence of malaria in the prone area could arise from various factors, e.g. wide use of pesticides in the agriculture sector and factors such as habitual patterns of movement of local people from problematic southeastern foci in Iran toward the Caspian Littoral. There are no new data on the resistance status of main malaria vectors in the Caspian Littoral, and this study was aimed at renewal data on conventional insecticides., Methods: The field strain of adult Anopheles superpictus and Anopheles maculipennis were collected using the hand catch method and transferred to the laboratory. The susceptibility tests were carried out against DDT 4%, Malathion 5%, Permethrin 0.75%, Deltamethrin 0.05%, and Lambda-cyhalothrin 0.05%, followed by the WHO's procedure., Results: The primary malaria vector in Caspian Littoral is An. maculipennis , revealed to be still resistant to DDT and mortality rate, LT 50 and LT 90 of female mosquitoes were 75.0%, 54.2, minutes and 111.3 minutes. The under 'verification required' status of An. maculipennis was also revealed to Lambda-cyhalothrin based on recent WHO's criteria. The malaria vector An. superpictus is also considered the second malaria vectors in the west parts of the studied area, which showed to be susceptible to all insecticides tested., Conclusion: DDT resistance is persisted in An. maculipennis despite stopping residual spraying with DDT since 1978 in the Caspian Littoral, but the occurrence of pyrethroid under 'verification required' status is a progressive threat to the possible development of cross-resistance in the future., (Copyright © 2021 The Authors. Published by Tehran University of Medical Sciences.)

Wild rats are known to carry different microorganisms and are considered a reservoir of zoonotic pathogens worldwide. The urban rats were collected from five districts of Tehran and Gram-negative bacteria (GNB) were isolated from fecal samples and were identified using classical biochemical tests. The antibiotic susceptibility patterns of isolated bacteria were determined by Kirby-Bauer disk diffusion method, the results of which were interpreted in line with CLSI guideline. The frequency of antibiotic-resistant genes was identified using multiplex-PCR. Moreover, PCR method was used to identify the frequency of Escherichia coli O157:H7 and main categories of diarrheagenic E. coli including EPEC, ETEC, EIEC, EAEC, and STEC pathotypes. A total of 100 Rattus norvegicus were trapped and fecal samples were collected. Overall, 72 fecal samples were positive for GNB. E. coli (n = 46/72) had the highest frequency among the isolated GNB. Among E. coli isolates, the highest and lowest resistance rates belonged to ampicillin (56.5%) and ceftriaxone (0%), respectively. Klebsiella spp. was 100% resistant to imipenem, and streptomycin (0%) was the most effective antimicrobial agent on Klebsiella spp. Among surveyed genes, bla TEM (95.8%) and bla aadA-1 (58.3%) had the highest frequency, while bla KPC , and bla CMY-2 were not detected among Enterobacteriaceae. Herein, O157: H7 serotype was not detected and aEPEC (87%) was the most common pathotype detected. Results suggested that rodents might be a reservoir of antimicrobial-resistant pathogens and rodent control along with implementation of surveillance programs should be considered as a critical priority for urban health.

Many studies have been done on the various factors affecting resistance to insecticides. The relationship between Wolbachia bacteria and resistance to insecticides is one of the factors that has attracted a lot of attentions. Wolbachia are obligatory intracellular endosymbionts that naturally occur in a wide range of arthropods and nematodes, including the mosquito Culex quinquefasciatus . Initially, the presence of bacteria was proved by molecular assays. Then the resistance level of this species was evaluated in adults against DDT 4.0% and deltamethrin 0.05% using the standard WHO guideline. After elimination of Wolbachia by tetracycline and its proof by molecular assays, the susceptibility tests were conducted again on uninfected line. Finally, the two lines were compared in terms of responding to insecticides. The findings indicated that there is no significant correlation between susceptibility of two lines in response to DDT 4.0% while they represented a significant correlation for deltamethrin (P =0.00). We propose that Wolbachia bacteria increase the susceptibility to deltamethrin but they show neutral effect on DDT susceptibility in Cx. quinquefasciatus . However, more studies on other vectors and insecticides still need to be done., Competing Interests: The authors declare no conflict of interest., (© 2021 Published by Elsevier Ltd.)

Background: Collecting live sand flies from indoor sites is a major challenge for researchers in large cities due to the reluctance of families to survey their homes. This study was conducted to assess the efficacy of two methods for collecting sand flies for use in susceptibility tests in the urban area of Kerman, southeastern Iran., Methods: Sandflies were mainly collected using both baited traps and hand catch methods from outdoor and indoor sites. Susceptibility tests were separately done according to the standard World Health Organization testing protocol on Phlebotomus sergenti , including 60-minute exposure to DDT 4.0%, propoxur 0.1%, deltamethrin 0.05%, and malathion 5.0%., Results: During this research, the natural habitats and suitable indoor sites were selected to predict the density of live sand fly with perfect accuracy. The number of live Ph. sergenti caught by hand catch and baited traps methods was 42 and 361 in indoor and outdoor sites, respectively. The mortality rate of Ph. sergenti exposed to DDT 4%, deltamethrin 0.05, malathion 5%, and propoxur was 100%., Conclusion: The baited traps showed a significant efficiency compared to hand catch for collecting live Ph. sergenti for use in susceptibility tests in urban areas. The Ph. sergenti collected from both indoor and outdoor sites were susceptible to all insecticides., (Copyright © 2021 The Authors. Published by Tehran University of Medical Sciences.)

Background: The aims of this study were to evaluate the efficiency of two capture methods for providing live sandflies used for determining the susceptibility level of Phlebotomus papatasi , the main vector of zoonotic cutaneous leishmaniasis in Lorestan Province, west of Iran., Methods: The sand flies were collected from indoor and outdoor by hand-catch and baited traps during the peak of seasonal activity. The susceptibility level of sand flies was assessed using insecticide-impregnated papers against DDT 4%, bendiocarb 0.1%, permethrin 0.75%, deltamethrin 0.05%, and cyfluthrin 0.15%., Results: A total of 2486 live sandflies were caught from both indoor and outdoor places. Totally 849 sand flies were caught from outdoors with a sex ratio(SR) 0.1 versus 1637 sand flies collected from indoor using the hand-catch method with SR= 0.6. The dominant species of sand flies was Ph. papatasi in the study area. Mortality rates of outdoor-collected sand flies were exposed to DDT 4%, deltamethrin 0.05%, permethrin 0.75%, and bendiocarb 0.1%, and mortality rate ranged from 92.0-97.9% and for indoor-collected sand flies were 87.7-96.8%. Both outdoor and indoor collected sand flies were susceptible to cyfluthrin 0.15% that caused 100% mortality., Conclusion: Based on the findings, the most appropriate method for collecting the live female Ph. papatasi is the baited traps due to providing enough females is necessary for conducting the susceptibility tests. The finding indicated that Ph. papatasi was resistant to DDT, under 'verification required' status to deltamethrin, permethrin, bendiocarb, and susceptible to cyfluthrin., (Copyright © 2020 The Authors. Published by Tehran University of Medical Sciences.)

Purpose: In southern Iran with a tropical climate, the above and underground cisterns of drinking water are the main habitats for immature stages of mosquitoes. The local people do not agree with using calcium hypochlorite granules in the water cisterns due to the change of water taste. Following the increased worldwide interest in using ultrasound technology for water disinfection, this study was conducted to assess the possible dual effects of ultrasound waves against immature mosquitoes. Materials and methods: Combinations of four power levels, two frequencies, three temperatures, and eight exposure times were applied to immature Culex pipiens . The exposure chamber of immature mosquitoes was the disposable cups placed inside a sonication bath containing tap water. Results: The mortality rate of immature mosquitos was 85-91% at 10-25 W, 30 °C and 130 kHz. The most effective ultrasound regimen for causing immature mortality included a fixed frequency of 130 kHz, a temperature of 30 °C, and a power level of 20-25 W. The lowest efficacy was observed in the egg stage. The median lethal time (LT 50 ) and ninety percent lethal time (LT 90 ) of sonication was 5.9 ± 0.6 min and 1.6 ± 0.1 min at 30 °C and a fixed frequency of 130 kHz. Conclusions: Evidence indicated effective ultrasound irradiation against immature mosquitoes in the water. This study provides a clear scientific basis for larvicidal properties induced by ultrasound emissions at laboratory conditions. The findings could be used for setting the ultrasound devices in the field investigations for achieving desired effectiveness.

Background: During the past decade, rapid development of insecticide resistance have been reported among many species of mosquito vectors against four main categories of insecticides worldwide. The aim of the research was to assess the variation trend of susceptibility levels of Culex quinquefasciatus to two insecticides separately for the field population compared with subsequent generations of the same sample after multiple colonization., Methods: Larvae and pupae of Cx. quinquefasciatus were collected from house sewages and reared to adult which blood-fed on roosters. Ten percent sucrose fed female mosquitoes aged 2-3 days were used for susceptibility tests with DDT and deltamethrin. Susceptibility levels was assessed in the adult stage of field stran Cx. quinquefasciatus against DDT 4.0% and deltamethrin 0.05% and continued up to next six generations undergoing multiple rearing at insectary condition., Results: The susceptibility levels to DDT 4.0% did not change compared to the field with the lab population to six generations. Regarding deltamethrin 0.05%, no significant difference was shown between field strain (58.3%) and 3 rd generation (52.7%) compared to the 6 th one (33.8%)., Conclusion: This finding may reflect the role of the kdr gene in resistance to organochlorine which has cross-resistance with pyrethroid insecticides. The results of this study clearly showed the irreversible trend of pyrethroid resistance among colonized mosquitoes. This is the first study of the resistance status of Cx. quinquefasciatus in Iran., (Copyright© Iranian Society of Medical Entomology & Tehran University of Medical Sciences.)

Seven genera and more than 98 species of mosquitoes (Diptera: Culicidae) are found in southwestern Asia (Harbach 1988). The most recent checklist of Iranian mosquitoes includes 69 species representing either seven or 11 genera depending on the classification used for aedines (Azari-Hamidian et al. 2019). Orthopodomyia pulcripalpis (Rondani) was the most recent genus and species to be added to the Iranian mosquito fauna (Azari-Hamidian et al. 2017).

Introduction: Entomological surveys of ectoparasites and their hosts are an essential tool for assessing the risks of rodent-borne diseases transmitted to humans by arthropod vectors. Objectives: This study was carried out to update the epidemiological data of plague with respect to species compositions of the rodents and their ectoparasites at enzootic foci located in Kurdistan Province, Iran. Methods: The rodents' habitats were selected based on past records of plague and subclimates in each study district with especial attention to the vegetation type. The trapped rodents were anesthetized using a chloroform chamber, and the animals were then examined for ectoparasites by brushing their hair over a pan containing water. The ectoparasites were collected with a fine brush and preserved in 70% ethanol in screw cap tubes. Results: A total of 208 rodents were trapped from three districts. Taxonomic ranking of the rodents indicated that the specimens belonged to 2 suborders of Myomorpha and Sciuromorpha, 4 families (Muridae, Muscardinidae, Cricetidae, and Sciuridae), 7 genera, including Meriones , Apodemus , Mus , Sciurus , Microtus , and Dryomys , and 15 species. Out of 208 rodents, only 56 (26.9%) were infested with 22 species of ectoparasites. Totally, 312 ectoparasites were isolated from 56 rodents, including 12 flea species (54.5%), 6 mite species (27.3%), 3 tick species (13.6%), and one louse species (4.6%). Five species of fleas were recorded for the first time in Kurdistan Province, including Ctenophthalmus iranus persicus , Paraceras melis melis , Nosopsyllus iranus iranus , Paraceras sp., and Ctenophyllus spp. Conclusion: The finding revealed new records for rodents and ectoparasites in Kurdistan Province, as well as changes in dominant rodent species and their ectoparasites compared to previous studies. This phenomenon can influence the changes in the incidence of plague and its epidemiology.

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., Competing Interests: None

Background & Objectives: Insecticides are the most important means of controlling pests in Iran especially for Culex pipiens complex species. The rational use of insecticides largely depends on understanding the susceptibility levels of these species. The study was designed to determine the susceptibility levels of Cx. pipiens complex (field and insectary strains) to various insecticides in the city of Tehran., Methods: The mortality rates of the field strain of Cx. pipiens complex after different exposure times to DDT (4%), bendiocarb (0.1%), propoxur (0.1%), malathion (5%), fenitrothion (1.0%), permethrin (0.75%), deltamethrin (0.05%), lambda-cyhalothrin (0.05%), etofenprox (0.5%), and cyfluthrin (0.15%) were determined. The mortality rates at the lethal time 50% (LT 50 ) and lethal time 90% (LT 90 ) values were calculated by plotting the regression line using Microsoft Office Excel software., Results: The mortality rates of the Cx. pipiens complex after 1 h exposure to the diagnostic doses of DDT (4%), bendiocarb (0.1%), propoxur (0.1%), malathion (5%), fenitrothion (1.0%), permethrin (0.75%), deltamethrin (0.05%), lambda-cyhalothrin (0.05%), etofenprox (0.5%), and cyfluthrin (0.15%) were 12, 58, 54, 82, 54, 34, 49, 40, 17, and 44%, respectively. According to the WHO classification of susceptibility levels, both field and insectary strains of Cx. pipiens complex in Tehran were resistant to these insecticides., Interpretation & Conclusion: The results of this study showed that field Cx. pipiens complex is resistant to all the groups of insecticides used., Competing Interests: None

Background: Phlebotominae sand flies (Diptera: Psychodidae) are the vectors of leishmaniasis. There are different methods for sand fly collection with different performance. The purpose of the current study was to compare the effect of different traps for collection of Phlebotominae sand flies in three endemic leishmaniasis foci in North Khorasan Province, northeast of Iran., Methods: Sand flies were collected using seven different traps from three villages, three times each twenty days during peak periods of seasonal activity in 2016., Results: A total of 7253 sand flies were collected. The specimens belonged to19 species. Phlebotomus sergenti was the most predominant species in the study area. Light trap baited with Carbon dioxide (CLT) and sticky paper trap (SPT) caught 22.6% and 22.3% of sand flies respectively. Animal baited trap (ABT) and white Shannon trap (WST) caught significantly fewer sand flies than the other traps. The sex ratio was different by phlebotominae sand fly species and collection methods. The sex ratio was highest in SPT and lowest in black Shannon trap (BST). Species diversity and species richness in SPT were more than other traps., Conclusion: Our findings confirm that CLT and SPT are the most efficient sand fly collection methods. CLT is higher attractive for females and Phlebotomus genus and is an ideal method for monitoring the population of Phlebotomus genus during surveillance. SPT is an inexpensive, convenient and easy to be used to detect the presence of sand flies at low densities and provide a more realistic estimation of sand flies biodiversity., (Copyright© Iranian Society of Medical Entomology & Tehran University of Medical Sciences.)

Background: Malaria is the main vector-borne disease worldwide. There are several reports of insecticide resistant in malaria vectors worldwide due to using different insecticides. The aim of this study was to evaluate different native plant extortions against main malaria vector, Anopheles stephensi in Iran for choosing the appropriate plant for formulation and use for vector control., Methods: The larvae of An. stephensi were reared in insectary, extraction of plants were carried out at department of Pharmacology. The standard WHO method for biological tests was used for calculation of LC 50 and LC 90 . Probit regration lines were plotted for calculation of LC 50 and LC 90 ., Results: In this study several plants including: Mentha spicata , Cymbopogon olivieri , Azadirachta indica , Melia azedarach , Lagetes minuta , Calotropis procera , Eucalyptus camaldulensis , Cupressus arizonica , Thymus vulgaris , Lawsonia inermis , Cedrus deodara , Cionura erecta , Bunium persicum , Carum carvi , Artemisia dracunculus , Rosmarinus officinalis were used. Results showed that Mentha spicata and Eucalyptus camaldulensis , had the lowest and highest LC 50 respectively., Conclusion: Results indicated that Mentha spicata and Eucalyptus camaldulensis, had the lowest and highest LC 50 respectively. Several other plant extract also showed significant mortality. The formulation of these plants should be prepared and evaluate at the field condition against malaria vectors., (Copyright© Iranian Society of Medical Entomology & Tehran University of Medical Sciences.)

Background: Cutaneous leishmaniasis due to Leishmania major is an important public health problem in the world. Khuzestan Province is one of the main foci of zoonotic cutaneous leishmaniasis (ZCL) in the southwest of Iran. We aimed to predict the spatial distribution of the vector and reservoir(s) of ZCL using decision-making tool and to prepare risk map of the disease using integrative GIS, RS and AHP methods in endemic foci in Shush (plain area) and Khorramshahr (coastal area) counties of Khuzestan Province, southern Iran from Mar 2012 to Jan 2013., Methods: Thirteen criteria including temperature, relative humidity, rainfall, soil texture, soil organic matter, soil pH, soil moisture, altitude, land cover, land use, underground water depth, distance from river, slope and distance from human dwelling with the highest chance of the presence of the main vector and reservoir of the disease were chosen for this study. Weights of the criteria classes were determined using the Expert choice 11 software. The presence probability maps of the vector and reservoir of the disease were prepared with the combination of AHP method and Arc GIS 9.3., Results: Based on the maps derived from the AHP model, in Khorramshahr study area, the highest probability of ZCL is predicted in Gharb Karoon rural district. The presence probability of ZCL was high in Hossein Abad and Benmoala rural districts in the northeast of Shush., Conclusion: Prediction maps of ZCL distribution pattern provide valuable information which can guide policy makers and health authorities to be precise in making appropriate decisions before occurrence of a possible disease outbreak., (Copyright© Iranian Society of Medical Entomology & Tehran University of Medical Sciences.)

Background: West Nile virus (WNV) can cause a fatal disease in humans and it is mainly transmitted to people through the bites of infected mosquitoes. Vector control using insecticides is a very important goal. Study of Culex pipiens resistance towards several insecticides in the city of Tehran, Iran was evaluated., Methods: Adult females reared from field-caught larvae from southern part of Tehran and lab strain reared in the insectary of Tehran University of Medical Science were determined for resistant status by exposing to 4% DDT, 0.1% bendiocarb, 0.1% propoxur, 1% fenitrothion, 0.05% deltamethrin, 0.75% permethrin, 0.05% lambda-cyhalothrin, 0.5% etofenprox, 5% malathion and 0.15% cyfluthrin papers using the standard WHO susceptibility tests., Results: Results clearly showed resistance development of Cx. pipiens against tested insecticides. Mortalities of Cx. pipiens were less than 90% with high resistance, low knock down rate and knock down time (50%) observed against insecticides. DDT and Malathion showed the most and least lethal time (LT 50 ) values for the field strain. The results of the knockdown test showed that DDT and deltamethrin had the most and least knockdown times (50%) for the field strain, respectively, while DDT and lambda-cyhalothrin had the most and least knockdown times (50%) for the lab strain, respectively., Conclusion: Resistance to mentioned insecticides in Cx. pipiens is widely distributed in southern part of Tehran. Regular implementation of susceptibility test in Cx. pipiens mosquitoes will help local public health authorities to develop new and better control strategies., (Copyright© Iranian Society of Medical Entomology & Tehran University of Medical Sciences.)

Forensically important flesh flies (Diptera: Sarcophagidae) often are not morphologically distinguishable, especially at the immature stage. In addition, female flies are quite similar in general morphology, making accurate identifications difficult. DNA-based technologies, particularly mitochondrial DNA (mtDNA), have been used for species-level identification. The cytochrome oxidase subunits I and II (COI-COII) sequences of Iranian Sarcophagidae are still unavailable in GenBank. In this study as many as 648 (540 males and 106 females) fly specimens from family Sarcophagidae, representing 10 sarcophagid species, including eight forensically important species were collected from seven locations in five Iranian provinces. Of these, 150 male specimens were identified based on both morphology of male genitalia and DNA sequencing analysis. Sequence data from the COI-COII regions for 10 flesh fly species collected in Iran were generated for the first time. Digestion of COI-COII region by restriction enzymes RsaI, EcoRV, and HinfI provided distinct restriction fragment length polymorphism profiles among the species and can serve as molecular markers for species determination. Phylogenetic analysis represented that the COI-COII sequences are helpful for delimitation of sarcophagid species and implementation in forensic entomology. However, the application of the COI-COII fragment as a species identifier requires great caution and additional species and markers should be studied to ensure accurate species identification in the future., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Mosquitoes are very important vectors of diseases to human. We aimed to establish the first spatial database on the mosquitoes of Isfahan Province, central Iran, and to predict the geographical distribution of species with medical importance., Methods: Mosquito larvae were collected from eight counties of Isfahan Province during 2014. Collected data were transferred to a database in ArcGIS and the distribution maps were created. MaxEnt model and jackknife analysis were used to predict the geographical distribution of two medical important species, and to find the effective variables for each species., Results: Totally, 1143 larvae were collected including 6 species, Anopheles maculipennis s.l., An. superpictus s.l., An. marteri , Culex hortensis , Cx. theileri and Culiseta longiareolata . The area under curve in MaxEnt model was 0.951 and 0.873 rather 1 for An. maculipennis s.l. and Cx. theileri , respectively. Culex theileri had wider and more appropriate niches across the province, except for the eastern area. The environmental variable with highest gain was mean temperature of the wettest quarter for Cx. theileri and temperature seasonality for An. maculipennis . Culex theileri , An. maculipennis s.l. and An. superpictus , three important vectors of parasitic agents to humans, were collected in this study., Conclusion: The mosquito collected and mapped can be considered for transmission of malaria and filariasis in the region. Bearing in mind the results of niche modeling for vector species, more studies on vectorial capacity and resistance status to different insecticides of these species are recommended., (Copyright© Iranian Society of Medical Entomology & Tehran University of Medical Sciences.)

Background: Some mosquito species which belong to the Culex. pipiens complex are primary vectors for West Nile virus, Sindbis, Dirofilaria immitis , and many arboviruses. Knockdown resistance (kdr) mutations in the voltage-gated sodium channel (VGSC) gene of Cx. pipiens that is inherited, is one of the important threats for the efficacy of pyrethroids insecticides. Knockdown resistance (kdr) mutation, L1014F, is a well-defined mechanism of resistance to pyrethroids and DDT in many insect species. The aim of study was to determine the mechanisms of Insecticide resistance in this species., Methods: Specimens of Cx. pipiens , the major vector of West Nile virus, were obtained in Tehran, Iran by collecting larvae from polluted wastewater in Qarchak of Tehran. In 2016 Insecticide susceptibility tests were performed according to WHO methods with deltamethrin 0.05%. We focused on determination of this point mutation in the VGSC gene of Cx. pipiens by Real-time PCR., Results: Our results revealed high levels of resistance to deltamethrin 0.05%. The lethal times i.e. LT 50 and LT 90 for deltamethrin were 2.1530 and 8.5117 h respectively. The result of Real-time PCR confirmed the presence of resistant genotype in all the members of tested population. This study is the first report on kdr genotyping of Cx. pipiens from Tehran and our results on the VGSC gene in position L1014F confirmed the TTA to TTT nucleotide change., Conclusion: This finding will provide a clue for management of insecticide resistance in mosquito which are vectors of arboviruses and decision for replacement of novel approach for vector control., Competing Interests: Conflict of interest The authors declare no conflict of interest.

Background: The detection of insecticide resistance in natural populations of Anopheles vectors is absolutely necessary for malaria control. CDC bottle bioassay as a new tools has been employed for detecting the insecticide resistance. For a limit number of mosquito vectors, diagnostic doses and diagnostic times for some insecticides have already been determined using this new assay. For the first time in the area, susceptibility levels of Anopheles stephensi was done with DDT, deltamethrin, and bendiocarb using CDC bottle bioassay and compared results with WHO standard test method., Methods: Anopheles stephensi were collected in larvae stage from the cisterns of drinking water in Chabahar port which considered as old malaria foci, Sistan and Baluchistan province. The field collected larvae were colonized at the insectary of School of Public Health (SPH), Tehran University of Medical Science. The susceptibility tests were carried out on sugar fed female mosquitoes aged 2-3 days, against DDT 4%, bendiocarb 1% and deltamethrin 0.05% using WHO and CDC susceptibility methods. The mortality and knockdown rates, as well as the parameters of regression analysis, including LT 50 and LT 90 , was calculated separately for the WHO and CDC methods., Results: The 24h mortality rates of An. stephensi were 28.6% and 25.6% for DDT, 60.8% and 64.6% for bendiocarb and 100% for deltamethrin using both WHO and CDC assay at 30 and 60min respectively. The 50% lethal times (LT 50 ) were estimated 44.9 and 66.2min, 38.9 and 81.8min and 0.7 and 15.0min respectively using both WHO and CDC susceptibility tests., Conclusion: The similar results of susceptibility levels were shown for DDT, bendiocarb and deltamethrin. The lethal times (LT 50 ) showed significant difference using both WHO and CDC bioassay methods.

Many researchers have focused on controlling pest insects and vectors by natural products because of their low environmental pollution. The present study was conducted to evaluate the antioxidant and larvicidal activities of chloroform and methanol extracts of the leaves, fruits, roots, and isolated coumarin compounds (prantschimgin, oxypeucedanin, and 6-hydroxymethylherniarin) of Ferulago trifida from the Apiaceae family against Anopheles stephensi as one of the main malaria vectors. For insecticidal evaluation, A. stephensi larvae were exposed to different concentrations of the extracts and pure compounds (0.625-1280 ppm) according to the WHO protocol. The mortality percentages were measured 24 h after treatment and lethal concentration values were calculated. In addition, radical scavenging activities of the mentioned extracts and compounds were measured by the DPPH method. The methanol extract of fruits showed potent insecticidal properties with LC 50 and LC 90 values of 2.94 and 18.12 ppm, respectively. The chloroform extracts of the fruits and leaves were the second and third extracts with larvicidal effects. Among pure compounds, only oxypeucedanin showed moderate toxicity against A. stephensi with LC 50 and LC 90 values of 116.54 and 346.41 ppm, respectively. The antioxidant activities of the methanol extracts of leaves and fruits were stronger than other extracts with IC 50 values of 155.83 and 159.32 ppm, respectively. In conclusion, the methanol extract of F. trifida fruits can be used as a potent bio-insecticide in green control programs of mosquitoes, especially A. stephensi.

The mosquito Culex pipiens is the primary vector of Rift Valley fever, West Nile, encephalitis, and Zika viruses, and periodic lymphatic filariasis. Developing insecticide resistance in mosquitoes demands the development of new approaches to fight these diseases. Paratransgenesis and RNAi approaches by using engineered bacteria have been shown to reduce mosquito vector competence. Serratia-AS1 is a bacterium found in mosquitoes and was genetically modified for expression of antimalaria effector molecules that repress development of malaria parasites in mosquitoes. The aim of this study was to determine how a genetically marked Serratia strain expressing the mCherry fluorescent protein (mCherry-Serratia) affects the colonization potential, life span, blood feeding behavior, fecundity, and fertility of Cx. pipiens. mCherry-Serratia bacteria disseminated into larvae, pupae, and newly emerged adults and dramatically increased in numbers following a blood meal. The bacterium was transmitted to progeny, showing that it can extend horizontally, transstadially, and vertically through the mosquito population. The presence of mCherry-Serratia did not affect blood feeding behavior, survival rate, fecundity, and fertility of Culex mosquitoes. This is the first study to evaluate the effects of an engineered bacteria on the fitness of Cx. pipiens. Although challenges remain, such as producing engineered bacteria to secrete anti-pathogens associated with Cx. pipiens, introducing such bacteria into mosquito populations, our findings of minimal fitness cost caused by Serratia-AS1 bode well for the development of paratransgenesis and RNAi approaches., (© The Author(s) 2018. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Ticks are able to transmit important diseases to humans, including Rocky Mountain spotted fever, Q fever, Crimean-Congo haemorrhagic fever, summer Russian encephalitis, and relapsing fever., Aims: To determine the repellency effect of 1% flumethrin pour-on formulation against hard ticks., Methods: The concentration of flumethrin pour-on formulation was 1 mg/10 kg body weight and was administered on the dorsal midline from the head to the base of the tail. The livestock included cows, goats, oxen and sheep in 2 villages in Ardabil Province, Islamic Republic of Iran., Results: We studied 200 livestock comprising 5 age groups (< 2, 3-4, 5-6, 7-8 and >8 years). The main hard ticks identified were Hyalomma species (62.5%) and Rhipicephalus bursa (37.5%). In the treatment village, the maximum number of ticks per animal was 11.6 in oxen, 9.5 in sheep, 8.9 in goats and 8.6 in cattle. The repellency effect of flumethrin remained for 2 months., Conclusions: Flumethrin provided 2 months protection against hard ticks. Therefore, it could be used in the livestock industry. Control of ticks is important for prevention of disease transmission., (Copyright © World Health Organization (WHO) 2018. Some rights reserved. This work is available under the CC BY-NC-SA 3.0 IGO license (https://creativecommons.org/licenses/by-nc-sa/3.0/igo).)

Background: Plasmodium berghei is a rodent malaria parasite and has been very valuable means in the progress of our understanding of the essential molecular and cellular biology of the malaria parasites. Availability of hosts such as mice and vectors such as Anopheles stephensi has made this parasite a suitable system to study the parasite-host and vector-parasite relationships., Methods: This study was performed at Medical Entomology and Parasitology laboratories of the School of Public Health, Tehran University of Medical Sciences, Iran in 2016. The investigation was carried out to describe life cycle and parameters influencing maintenance of the parasite within the mice or the mosquito., Results: Results have revealed details and addressed some parameters and points influence maintenance of various life stages of the parasite including merozoites, macrogametocytes, ookinetes, oocysts and sporozoites in the laboratory model P. berghei-A. stephensi -BALB/c mouse. Injection of fresh infected blood results in higher gametocytemia in the animals. The more injected parasites result in earlier and higher parasitemia and exfelagellation centers in the mice blood. However, the highest number of infected mosquitoes and oocysts formation were observed when the parasitemia and exflagellation centers per microscopic field were 9% and 3.6 in the infected mice respectively. The infected mosquitoes should be maintained on 8% (w/v) fructose, 0.05% (w/v) PABA at 20±1 °C and 50%-80% relative humidity., Conclusion: This study helps to understand the biology of vertebrate-parasite and mosquito-malaria interactions that may aid in the development of a new generation of drug/vaccine and vector-based measures for malaria control., Competing Interests: Conflict of interest The authors declare that there is no conflict of interests.

Background: Malaria continues to be a main vector-borne public health problem in Iran. The endemic foci of the disease are mainly located in south-eastern part of the country. Iran is now launching the elimination of malaria. Studies on the bioecology and susceptibility of malaria vectors to insecticide are essential in this phase., Methods: The literature on bio-ecology of Anopheles superpictus s.l. was reviewed in Iran in more than half a century. Different aspects including, distribution, key identification, larval habitats, flight range, seasonal activities, irritability/susceptibility to insecticides, and anthropophilicity index were identified., Results: The adult females of An. superpictus s.l. were susceptible to all WHO-recommended imagicides except DDT. Distribution, morphology, sibling species, larval habitat, flight range, irritability tests, sustainability index, blood feeding preference and related factors were discussed in details., Conclusion: Results of the evaluating will help for decision making of authorities for vector control.

Background: Anopheles stephensi is a key urban malaria vector in the Indian subcontinent and Middle East including south and southeast of Iran. Wide application of insecticides resulted in resistance of this species to various insecticides in these regions. This study was conducted to reveal the role of metabolic mechanisms in the development of resistance in An. stephensi to DDT and cyfluthrin., Methods: Field mosquito specimens were collected from Chabahar Seaport, southeast corner of Iran, in 2015. Insecticide susceptibility and enzyme assays were conducted as recommended by WHO., Results: Mean enzyme ratios were 3.95 and 3.04 for α- esterases and 2.40 and 1.97 for β- esterases in the DDT and cyfluthrin- resistant populations correspondingly compared with the susceptible strain. The GSTs enzyme mean activity ratios were 5.07 and 2.55 in the DDT and cyfluthrin- resistant populations compared with the susceptible beech strain. The cytochrome p450s enzyme ratios were 1.11 and 1.28 in the DDT and cyfluthrin- resistant populations respectively compared with the susceptible beech strain., Conclusion: Metabolic mechanisms play a crucial role in the development of DDT and cyfluthrin resistance in An. stephensi , therefore, further evaluation of the mechanisms involved as well as implementation of proper insecticide resistance management strategies are recommended.

Background: We aimed to determine the species composition, richness and diversity indices of the Phlebotomine sand flies at different topographic condition in visceral (VL) and cutaneous leishmaniasis (CL) foci in the northeast of Iran., Methods: This cross-sectional study was conducted during 2016 in North Khorasan Province. The sampling was focused on rural regions, where human cases of VL and/or CL were diagnosed and reported during last 5 years. Sand flies were collected three times each twenty days during peak periods of seasonal activity. Seven collection methods were used. Some Alpha and Beta diversity indices were calculated., Results: Overall, 7253 sand flies were collected and identified. They were from 19 species of Phlebotominae sand-flies. Phlebotomus sergenti and Ph. papatasi were the most prevalent (84.9%) species in the study area. Species richness (S) was very different in three areas and were18, 8, and 4 respectively but Evenness (E) were 0.357, 0.345, and 0.380, so evenness was almost equal in the study areas. Shannon Index (H) and Margalef Richness Index were calculated 1.033, 0.718, 0.527 and 2.117, 0.8998, 0.4006 respectively., Conclusion: The sand fly fauna in North Khorasan Province was very rich and often included some of the most important proven or suspected vectors of leishmaniasis. Species diversity indices (Shannon index, and Simpson's index) were not high due to decreasing in evenness. The Margalef richness index could accurately reflect the biodiversity of sand flies between three subtidal locations.

Background: Scorpions have medical importance in the studied area with 2377 cases of envenoming during past six years. This study was the first to explore the scorpion species and dispersion in the Kazerun District during 2014-2015., Methods: The studied sites were selected based on different topographic conditions such as plain, foothill and mountainous which formed four geographical zones with three villages in each zone. The sampling was carried out twice each month throughout the year. Daytime collections were carried out using hand digging tools for moving stones and excavate the borrows, as well as night sampling, is done with the black light device. The coordinate of locations was recorded with a GPS. The collected specimens were maintained in 70% ethanol and identified using authorized keys., Results: Overall, 800 scorpions were sampled from different parts of Kazerun District, bringing the species richness to 9 belonged to 3 families of Buthidae, Scorpionidae, and Hemiscorpionidae. The Mesobuthus eupeus (84.6%) was prominent vice versa M. caucasicus (0.1%) had lowest abundance. Other species comprised Compsobuthus matthiesseni (5.3%), Androctonus crassicauda (5.0%), Razianus zarudnyi (2.0%), Hemiscorpius lepturus (1.5%), Orthochirus sp (0.9%), Hottentotta zagrosensis (0.4%), and Scorpio maurus (0.3%). The seasonal activity of the scorpions showed a lower peak in Mar, with the main peaks in Aug for the dominant species. Mesobuthus caucasicus is recorded for the first time in the Fars Province, southern Iran., Conclusion: All the known dangerous scorpions, including H. lepturus , M. eupeus and A. crassicauda were revealed in the studied area.

Zoonotic Cutaneous Leishmaniasis (ZCL) is one of the endemic diseases in central part of Iran. The aim of this cross-sectional study was to find the areas with a higher risk of infection considering the distribution of vector, reservoir hosts and human infection. Passive data recorded the positive cases of cutaneous leishmaniasis in Yazd province health center were collected for 10 years, from 2007 to 2016 at the County level. Considering all earlier studies conducted in Yazd province, records of Phlebotomus papatasi, the main vector of ZCL, and Rhombomys opimus, the main reservoir of ZCL, were collected and entered in a database. ArcGIS and MaxEnt model were used to map and predict the best ecological niches for both vector and reservoir. The most cumulative incidence of the disease was found to be in Khatam County, south of Yazd province. The area under curve (AUC) for R. opimus and P. papatasi was 0.955 and 0.914, respectively. We found higher presence probability of both vector and reservoir in central and eastern parts of the province. The jackknife test indicated that temperature and normalized difference vegetation index (NDVI) had the most effect on the model for the vector and reservoir, respectively. The areas with higher presence probability for the reservoirs and vectors were considered having the higher potential for ZCL transmission. These findings can be used to prevent and control the disease., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Background: Natural plant products as larvicides could be considered as desirable alternatives to synthetic chemical insecticides for vector management. This study was undertaken to assess the mosquito larvicide activity of the essential oil from fresh leaves of Platycladus orientalis against two medically important species of mosquito vectors., Methods: Essential oil was extracted by hydrodistillation and analyzed with gas chromatography and mass spectrometry (GC-MS). Fresh leaves of P. orientalis tree (500g) were collected in June 2014 from Tehran, Iran and was authenticated at the Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.In addition, the larvicidal potential of oil was evaluated against late-3 rd or young-4 th instar larvae of Anopheles stephensi and Culex pipiens under laboratory condition. The mortality counts were made after 24h and LC 50 and LC 90 values were calculated., Results: Forty-six components in leaves of P. orientalis were identified. The major components were α-Pinene (20.17%), 3-Carene (14%) and Cedrol (9.51%). The LC 50 values against An. stephensi and Cx. pipiens larvae were 11.67ppm and 18.60ppm after 24h, respectively., Conclusion: Platycladus orientalis oil could be considered as a natural larvicide for mosquito larval control.

Background: Scrophularia striata is a perennial plant which is native in all parts of Iran, Turkey, and Azerbaijan. In this study, the total phenol content, antioxidant and larvicidal activities of total extract and different fractions of this plant were evaluated., Methods: The aerial parts of S. striata were collected from Boli village, Illam Province, western Iran in Apr 2013. The total phenol content of total extract and different fractions were evaluated by Folin-Ciocalteu method. Moreover, antioxidant activity was tested by DPPH and FRAPS assays. Larvicidal activity was investigated according to standard method described by WHO., Results: Ethyl acetate fraction (EF) had the highest content of total phenol (75.9±0.06mg Gallic acid equivalent/g dry extract). Furthermore, among the tested extract, methanol-water fraction (MWF), total methanol extract (TME) and water fraction (WF) showed the highest antioxidant activity in the DPPH assay (IC 50 = 226.8, 283.66 and 299.4 μg.ml -1 , respectively). In FRAP assay MWF and WF and TME had the highest antioxidant activities (664.4±0.002, 565.3±0.003, 519.5±0.003mmol FeII/g dry extract, respectively). Ethyl acetate fraction had maximum larvicidal activity (LC 50 49.1ppm) followed by TME (LC 50 64.26ppm) and hexane fraction (HF) (LC 50 89.69)., Conclusion: Scrophularia striata collected from west of Iran illustrated considerable antioxidant and larvicidal effects and further in vitro and in vivo experimental models for investigation would be required.

Cutaneous leishmaniasis (CL) is endemic and a major health problem in 17 provinces out of 31 in Iran. This study aimed to determine vectors and reservoirs of the disease using molecular techniques in the borderline of Iran and Iraq. Sand flies and rodents were sampled using sticky paper traps and metal wire live traps, respectively, in the selected villages. About 10% of archived confirmed human positive slides was randomly checked for Leishmania by PCR-RFLP assay. The female sand flies were dissected in alcohol 96% in a sterile condition, the head and two segments of the abdomen end permanently mounted for identification and the remaining of body used for DNA extraction. The direct parasitological tests were carried out on the stained slides of rodents for Leishmania as well as PCR-RFLP assay used for molecular detection of parasite. A total of 2050 sand flies were identified comprising of Phlebotomus papatasi , Sergentomyia sintoni , Se . clydei , Se . mervynae , Se . theodori, Se . dentate and Se . iranica . The Ph . papatasi was ranked as a prevailing sand fly species. Molecular tests on female sand flies revealed infection of Ph . papatasi to Leishmania major . Direct parasitology and molecular tests confirmed of 20% infection to L . major among the sole rodents species " Tatera indica ". Due to wide dispersion of rodents colonies in the area and long favorite climate condition for sand flies, the CL foci will be provided the health risk for the religious tourists., Competing Interests: Compliance with ethical standardsThe authors declare that they have no conflict of interest.

Background: Culex pipiens is a mosquito species distributed in different parts of Iran. It is known as vector of some diseases as well as a nuisance insect. A successful control campaign needs to study the biology, ecology and susceptibility status of the target pest. The aim of this study was to investigate the susceptibility status of Culex pipiens to some insecticides recommended by WHO in north of Iran., Methods: Larvae were collected from different breeding places in Sari County by standard dipping method. They were reared to adult stage in an under standard condition. World Health Organization (WHO) standard test kits and the diagnostic dose of 12 imagicides were used for adults, while two larvicides were used against larvae., Results: The least and highest mortality rates after exposure to insecticides were 4.7% (Propoxur 0.1%) and 76.4% (Etofenprox 0.5%), respectively. Larvae showed high resistance to temephos (19.19%) and malathion (54.63%). Culex pipiens was found to be resistant to all used insecticides/larvicides at diagnostic dose., Conclusion: Intensive use of pesticides against agriculture and urban pests increased the resistance level of this species to different insecticides, although currently there is no control program against it. Study on the mechanisms of resistant in this species to different insecticides is recommended.

Background: Malaria, a mosquito-transmitted disease, is still a major human health problem all over the world. Larviciding is a component of comprehensive control program to overcome the disease. Negative aspects of synthetic insecticides application, such as environmental safety concerns, have favored use of natural insecticides., Methods: Larvicidal activity of essential oil, extracts and fractions of a wild grown and a cultivated type of Bunium persicum fruits against malaria vector Anopheles stephensi was assessed according to the method described by WHO., Results: Bunium persicum showed remarkable potency against An. stephensi larvae. LC 50 values for essential oil, total extract, petroleum ether fraction and methanol fraction were 27.4284, 64.9933, 85.9933 and 255.7486ppm for wild type, and 21.3823, 63.2580, 62.7814 and 152.6357ppm for cultivated one., Conclusion: The results of this study suggest B. persicum as a valuable source of natural insecticides against malaria vector Anopheles stephensi .

Background: The aim of this study was to evaluate different concentrations of deltamethrin combined with formulated piperonyl butoxide (PBO) synergist on various surfaces against the wild strain of Anopheles stephensi , the main malaria vector in Southern Iran under semi-field condition., Methods: Four concentrations of deltamethrin WG 25% (Tagros) and PBO 800EC-UV (Endura) were prepared and sprayed on the pre-designed surfaces in accordance with WHO alliance line of the IRS Micronair®. The WHO's recommended bioassay kit and method was used during this study., Results: Comparing the mortality rate of mosquitoes, the results showed a significant difference between months after treatment of IRS (Indoor Residual Spraying) (P< 0.05) but didn't show any significant differences between days during the first and second months (P> 0.05).Statistical test revealed a significance difference between mortality rate of mosquitoes in exposing to concentrations of 1 and 4 (P< 0.05) which demonstrated effect of synergizing PBO on mortality rate., Conclusion: This research as the first semi-field trial on deltamethrin added to different concentrations of formulated PBO for IRS, indicates that deltamethrin+10X PBO is more effective than other concentrations. Therefore, using synergists can be suggested as a new tool for prevention of pyrethriod resistance, although more studies are recommended.

Background: Enterobacter cloacae bacterium is a known symbiont of the most Anopheles gut microflora and nominated as a good candidate for paratransgenic control of malaria. However, the population dynamics of this bacterium within An. stephensi and its introduction methods to the mosquitoes have not yet been explored., Methods: Enterobacter cloacae subsp. dissolvens expressing green fluorescent protein and defensin (GFP-D) was used to study transstadial transmission and the course of time, larval habitat, sugar, and blood meal on dynamics of the bacterium in the mosquito life stages in the laboratory condition. The bacterial quantities were measured by plating samples and counting GFP expressing colonies on the Tet-BHI agar medium., Results: The E. cloacae population remained stable in sugar bait at least for eleven days whereas it was lowered in the insectary larval habitat where the bacteria inadequately recycled. The bacterium was weakly transmitted transstadially from larval to adult stage. The bacterial populations increased smoothly and then dramatically in the guts of An. stephensi following sugar and blood meal respectively followed by a gradual reduction over the time., Conclusion: Enterobacter cloacae was highly stable in sugar bait and increased tremendously in the gut of female adult An. stephensi within 24h post blood meal. Sugar bait stations can be used for introduction of the transgenic bacteria in a paratransgenic approach. It is recommended to evaluate the attraction of sugar bait in combination with attractive kairomones as well as its stability and survival rate in the semi-field or field conditions.

Members of the Culex (Culex) pipiens assemblage are known vectors of deadly encephalitides, periodic filariasis, and West Nile virus throughout the world. However, members of this assemblage are morphologically indistinguishable or hard to distinguish and play distinct roles in transmission of the diseases. The current study aimed to provide further evidence on utility of the two most popular nuclear (ITS2-rDNA) and mitochondrial (COI barcode region) genetic markers to identify members of the assemblage. Culex pipiens assemblage specimens from different climate zones of Iran were collected and identified to species level based on morphological characteristics. Nucleotide sequences of the loci for the specimens plus available data in the GenBank were analyzed to find species specific genetic structures useful for diagnosis purposes. ITS2 region was highly divergent within species or populations suggesting lack of consistency as a reliable molecular marker. In contrast, sequence analysis of 710 bp of COI gene revealed three fixed haplotypes named here "C, T, H" within the assemblage which can be distinguished by HaeIII and AluI enzymes. There were a correlation between the haplotypes and the world climate regions, where the haplotypes H/T and C are present mainly in temperate and tropical regions of the world, respectively. In the New world, Australia, and Japan only haplotype H is found. In conjunction between tropical and temperate regions such Iran, China, and Turkey, a mix of C/H or C/H/T are present. Although, the haplotypes are not strictly species-specific, however, Cx. quinquefasciatus was mainly of haplotype C. Due to the lack of mating barrier and questionable taxonomic situation of the complex members, the mentioned haplotypes in combination with other morphological and molecular characters might be used to address the genetic structure of the studied populations., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Background: We aimed to investigate different biological properties of aerial parts essential oil of Ferulago trifida Boiss and larvicidal activity of its volatile oils from all parts of plant., Methods: Essential oil was prepared by steam distillation and analyzed by Gas chromatography and GC/Mass. Antioxidant, antimicrobial, cytotoxic effects and AChE inhibitory of the oil were investigated using DPPH, disk diffusion method, MTT assay and Ellman methods. Larvicidal activity of F. trifida essential oil against malaria vector Anopheles stephensi was carried out according to the method described by WHO., Results: In GC and GC/MS analysis, 58 compounds were identified in the aerial parts essential oil, of which E-verbenol (9.66%), isobutyl acetate (25.73%) and E-β-caryophyllene (8.68%) were main compounds. The oil showed (IC 50 = 111.2μg/ml) in DPPH and IC 50 = 21.5 mg/ml in the investigation of AChE inhibitory. Furthermore, the oil demonstrated toxicity with (LD 50 = 1.1μg/ml) in brine shrimp lethality test and with (IC 50 = 22.0, 25.0 and 42.55 μg/ml) on three cancerous cell lines (MCF-7, A-549 and HT-29) respectively. LC 50 of stem, root, aerial parts, fruits, and flowers essential oils against larvae of An. stephensi were equal with 10.46, 22.27, 20.50, 31.93 and 79.87ppm respectively. In antimicrobial activities, essential oil was effective on all specimens except Escherichia coli , Aspergillus niger and Candida albicans., Conclusion: The essential oil showed moderate antioxidant activity, strong antimicrobial properties and good toxic effect in brine shrimp test and MTT assay on three cancerous cell lines.

Objective: To determine the larvicidal activities of petroleum ether, chloroform, ethyl acetate and methanol fractions of roots and fruits extracts of Astrodaucus persicus from Apiaceae family against malaria vector, Anopheles stephensi (An. stephensi)., Methods: Twenty five third instar larvae of An. stephensi were exposed to various concentrations (10-160 g/L) of fractions and were assayed according to World Health Organization protocol. The larval mortality was calculated after 24 h treatment., Results: Among tested fractions, the highest larvicidal efficacy was observed from ethyl acetate fraction of fruits extract with 50% and 90% mortality values (LC 50 and LC 90 ) of 34.49 g/L and 108.61 g/L, respectively. Chloroform fraction of fruits extract was the second larvicidal sample with LC 50 of 45.11 g/L and LC 90 of 139.36 g/L. Petroleum ether fractions of fruits and roots and methanol fraction of fruits showed moderate toxicity against An. stephensi., Conclusions: Astrodaucus persicus is a potential source of valuable and natural larvicidal compounds against malaria vector, An. stephensi and can be used in mosquitoes control programs as an alternative to synthetic insecticides., (Copyright © 2017 Hainan Medical University. Production and hosting by Elsevier B.V. All rights reserved.)