Search

Background While Iran is on the path to eliminating malaria, the disease with 4.9 million estimated cases and 9300 estimated deaths in 2018 remains a serious health problem in the World Health Organization (WHO) Eastern Mediterranean Region. Anopheles stephensi is the main malaria vector in Iran and its range extends from Iraq to western China. Recently, the vector invaded new territories in Sri Lanka and countries in the Horn of Africa. Insecticide resistance in An. stephensi is a potential issue in controlling the spread of this vector. Methods Data were collated from national and international databases, including PubMed, Google Scholar, Scopus, ScienceDirect, SID, and IranMedex using appropriate search terms. Results Indoor residual spaying (IRS) with DDT was piloted in Iran in 1945 and subsequently used in the malaria eradication programme. Resistance to DDT in An. stephensi was detected in Iran, Iraq, Pakistan, and Saudi Arabia in the late 1960s. Malathion was used for malaria control in Iran in 1967, then propoxur in 1978, followed by pirimiphos-methyl from 1992 to 1994. The pyrethroid insecticide lambda-cyhalothrin was used from 1994 to 2003 followed by deltamethrin IRS and long-lasting insecticidal nets (LLINs). Some of these insecticides with the same sequence were used in other malaria-endemic countries of the region. Pyrethroid resistance was detected in An. stephensi in Afghanistan in 2010, in 2011 in India and in 2012 in Iran. The newly invaded population of An. stephensi in Ethiopia was resistant to insecticides of all four major insecticide classes. Different mechanisms of insecticide resistance, including metabolic and insecticide target site insensitivity, have been developed in An. stephensi. Resistance to DDT was initially glutathione S-transferase based. Target site knockdown resistance was later selected by pyrethroids. Esterases and altered acetylcholinesterase are the underlying cause of organophosphate resistance and cytochrome p450s were involved in pyrethroid metabolic resistance. Conclusions Anopheles stephensi is a major malaria vector in Iran and many countries in the region and beyond. The species is leading in terms of development of insecticide resistance as well as developing a variety of resistance mechanisms. Knowledge of the evolution of insecticide resistance and their underlying mechanisms, in particular, are important to Iran, considering the final steps the country is taking towards malaria elimination, but also to other countries in the region for their battle against malaria. This systematic review may also be of value to countries and territories newly invaded by this species, especially in the Horn of Africa, where the malaria situation is already dire.

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 Due to the effect of synthetic and commercial insecticides on non-target organisms and the resistance of mosquitoes, non-chemical and environmentally friendly methods have become prevalent in recent years. The present study was to isolate entomopathogenic fungi with toxic effects on mosquitoes in natural larval habitats. Methods Larvae of mosquitoes were collected from Central, Qamsar, Niasar, and Barzok Districts in Kashan County, Central Iran by standard dipping method, from April to late December 2019. Dead larvae, live larvae showing signs of infection, and larvae and pupae with a white coating of fungal mycelium on the outer surface of their bodies were isolated from the rest of the larvae and sterilized with 10% sodium hypochlorite for 2 min, then washed twice with distilled water and transferred to potato-dextrose-agar (PDA) and water-agar (WA) media and incubated at 25 ± 2 °C for 3–4 days. Larvae and fungi were identified morphologically based on identification keys. Results A total of 9789 larvae were collected from urban and rural areas in Kashan County. Thirteen species were identified which were recognized to belong to three genera, including Anopheles (7.89%), Culiseta (17.42%) and Culex (74.69%). A total of 105 larvae, including Anopheles superpictus sensu lato (s.l), Anopheles maculipennis s.l., Culex deserticola, Culex perexiguus, and Culiseta longiareolata were found to be infected by Nattrassia mangiferae, Aspergillus niger, Aspergillus fumigatus, Trichoderma spp., and Penicillium spp. Of these, Penicillium spp. was the most abundant fungus isolated and identified from the larval habitats, while An. superpictus s.l. was the most infected mosquito species. Conclusions Based on the observations and results obtained of the study, isolated fungi had the potential efficacy for pathogenicity on mosquito larvae. It is suggested that their effects on mosquito larvae should be investigated in the laboratory. The most important point, however, is the proper way of exploiting these biocontrol agents to maximize their effect on reducing the population of vector mosquito larvae without any negative effect on non-target organisms.

Using molecular techniques and bioinformatics tools, we studied the vector-host interactions and the molecular epidemiology of West Nile virus (WNV) in western Iran. Mosquitoes were collected during 2017 and 2018. DNA typing assays were used to study vector-host interactions. Mosquitoes were screened by RT-PCR for the genomes of five virus families. WNV-positive samples were fully sequenced and evolutionary tree and molecular architecture were constructed by Geneious software and SWISS-MODEL workspace, respectively. A total of 5028 mosquito specimens were collected and identified. The most prevalent species was Culex (Cx.) pipiens complex (57.3%). Analysis of the blood-feeding preferences of blood-fed mosquitoes revealed six mammalian and one bird species as hosts. One mosquito pool containing non-blood-fed Cx. theileri and one blood-fed Culex pipiens pipiens (Cpp.) biotype pipiens were positive for WNV. A phylogram indicated that the obtained WNV sequences belonged to lineage 2, subclade 2 g. Several amino acid substitutions suspected as virulence markers were observed in the Iranian WNV strains. The three-dimensional structural homology model of the E-protein identified hot spot domains known to facilitate virus invasion and neurotropism. The recent detection of WNV lineage 2 in mosquitoes from several regions of Iran in consecutive years suggests that the virus is established in the country.

Background: Southern part of the country is a high risk for mosquito transmitted Arboviruses. This study was carried out to determine the base line susceptibility of the Aedini mosquitoes to the WHO-recommended insecticide. Methods: Larval collection was carried out by dipping method and adult collection occurred by suction tube from January to December 2017. The adult susceptibility test was assessed to Bendiocarb 0.1%, DDT 4%, Deltamethrin 0.05%, Lambda-cyhalothrin 0.05%, Malathion 5% and, Permethrin 0.75% at different interval times as well as at discriminative dose recommended by WHO. The larval susceptibility test was occurred using Temephos and Bacillus thuringiensis serotype H-14, at different concentrations. The LT50, LT90 and LC50, LC90 values were calculated for plotting the regression line using Microsoft office Excel software ver. 2007. Results: Aedes caspius was quite resistant to DDT, Malathion, Bendiocarb and showed susceptible or tolerant to other insecticides.The LT50 and LT90 values to DDT in this species were 157.896, and 301.006 minutes, respectively. The LC50 and LC90 values of Ae. caspius to Temephos were 0.000068, and 0.000130ppm, the figures for B. thuringiensis was 111.62 and 210.2ppm, respectively. Conclusion: A routine and continuous study for monitoring and evaluation of different species of Aedes to insectides is recommend at different parts of country for decision making.

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.

Aedes caspius has a wide distribution throughout the world and can transmit Chikungunya virus, West Nile Virus (WNV), Tahyna virus and the bacterium Francisella tularensis. Sequences of the mitochondrial cytochrome C-oxidase subunit 1 (COI) and cytochrome C-oxidase subunit 2 (COII) genes have been widely used to estimate phylogenetic relationships at different taxonomic levels among this species. Adult collections were carried out by human bait, Center for Deseases Control Light Traps (CDC-LT) and aspirator during February/April, June and October/December 2013-2015 from different southern provinces of Iran and then identified morphologically with reliable keys. A total of 3,570 adult mosquitoes were collected and identified as belonging to three genera, including five species of Culex, six species of Aedes and one species of Culiseta. In this study, 1,796 specimens of Aedes caspius were identified from four provinces. Based on the COI and COII sequences obtained for Ae. caspius population, 12 and 11 haplotypes were identified, respectively. The present study evidenced a high degree of intraspecific variation among these populations of Ae. caspius.

Ticks serve as important vectors of some pathogens of medical importance all over the world and identification of their rate of infection plays an important role for further control of diseases. In the current study, we investigated on ticks collected from north of Iran where raising and caring livestock are the main task of the people in order to find evidences of infection of Babesia, Theileria, Anaplasma and Ehrlichia microbial agents. Totally, 609 hard tick species from two genera Hyalomma and Rhipicephalus including; Hy. scupense, Hy. dromedarii, Hy. rufipes, Hy. marginatum, Hy. asiaticum, Hy. anatolicum, R. bursa, R. sanguineus and R. turanicus were identified. Molecular analysis revealed the presence of Anaplasma, Ehrlichia, Babesia and Theileria microorganism agents in all collected tick species except Hy. asiaticum and R. turanicus. To the best of our knowledge, this is the first report on identification of B. occultans in Hyalomma anatolicum and B. ovis in Hyalomma sp in Iran.

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.

Objective Screening of mosquitoes for viruses is an important forecasting tool for emerging and re-emerging arboviruses. Iran has been known to harbour medically important arboviruses such as West Nile virus (WNV) and dengue virus (DENV) based on seroepidemiological data. However, there are no data about the potential mosquito vectors for arboviruses in Iran. This study was performed to provide mosquito and arbovirus data from Iran. Materials and Methods A total of 32 317 mosquitos were collected at 16 sites in five provinces of Iran in 2015 and 2016. RT-PCR for detection of flaviviruses was performed. The PCR amplicons were sequenced, and 109 WNV sequences, including one obtained in this study, were used for phylogenetic analyses. Results The 32 317 mosquito specimens belonging to 25 species were morphologically distinguished and distributed into 1222 pools. Culex pipiens s.l. comprised 56.429%. One mosquito pool (0.08%), containing 46 unfed Cx. pipiens pipiens form pipiens (Cpp) captured in August 2015, was positive for flavivirus RNA. Subsequent sequencing and phylogenetic analyses revealed that the detected Iranian WNV strain belongs to lineage 2 and clusters with a strain recently detected in humans. No flaviviruses other than WNV were detected in the mosquito pools. Conclusion Cpp could be a vector for WNV in Iran. Our findings indicate recent circulation of WNV lineage-2 strain in Iran and provide a solid base for more targeted arbovirus surveillance programs.

Phlebotomus caucasicus Marzinovsky and Phlebotomus mongolensis Sinton are morphologically similar sand fly species. Finding a reliable, fast, and simple method to differentiate these two sand flies is important in understanding their role in the transmission of Leishmania parasite. In our study, 20 specimens of male P. caucasicus, 4 specimens of male P. mongolensis, and 16 specimens of female of both species (Caucasicus group) were examined by polymerase chain reaction (PCR). The result shows identical patterns with a visible fragment of about 500 bp in size. In restriction fragment length polymorphism (RFLP), we observed identical patterns with TasI used as the restriction enzyme. After alignment with sequences of the ITS2 partial gene in GenBank, a perfect match was obtained for the P. mongolensis, but not for P. caucasicus whose sequence was not present in the GenBank. Based on the results of our study, the RFLP-PCR method with nucleotide gene fragment ITS2 was a rapid and reliable method for differentiating these sand fly species.

Malaria is considered as a major public health problem in southern areas of Iran. The goal of this study was to predict best ecological niches of three main malaria vectors of Iran: Anopheles stephensi , Anopheles culicifacies s.l. and Anopheles fluviatilis s.l. A databank was created which included all published data about Anopheles species of Iran from 1961 to 2015. The suitable environmental niches for the three above mentioned Anopheles species were predicted using maximum entropy model (MaxEnt). AUC (area under Roc curve) values were 0.943, 0.974 and 0.956 for An. stephensi , An. culicifacies s.l. and An. fluviatilis s.l respectively, which are considered as high potential power of model in the prediction of species niches. The biggest bioclimatic contributor for An. stephensi and An. fluviatilis s.l. was bio 15 (precipitation seasonality), 25.5% and 36.1% respectively, followed by bio 1 (annual mean temperature), 20.8% for An. stephensi and bio 4 (temperature seasonality) with 49.4% contribution for An. culicifacies s.l. This is the first step in the mapping of the country’s malaria vectors. Hence, future weather situation can change the dispersal maps of Anopheles . Iran is under elimination phase of malaria, so that such spatio-temporal studies are essential and could provide guideline for decision makers for IVM strategies in problematic areas.

Background Global warming and climate change affect various aspects of mankind, including public health. Anopheles mosquitoes are of Public Health importance and can be affected by global warming and other environmental variables. Here, we studied the distribution of Anopheles vectors of malaria in relation to environmental variables in Iran. Methods Long-term meteorological and entomological data of about 50 years in retrospect were collected and arranged in a geo-database and analyzed using ArcGIS ver. 9.3 and exported to SPSS ver. 20 for statistical analysis. Results Distribution maps have been updated for seven species of Anopheles vectors of malaria which involved Anopheles culicifacies s.l., An. fluviatilis s.l., An. stephensi, An. dthali, An. sacharovi, An. maculipennis.l. and An. superpictus in Iran. Distribution maps of vectors were made based on district areas using Kriging model. Historical and recent records were demonstrated for each Anopheles based on climatic factors in the distribution areas of each Anopheles vectors. Discussion Iran, like other parts of the world is faced with warming and this probably affected the distribution of Anopheles vectors. Despite the warming phenomenon, the country’s climate had changed during the cold season as temperatures became colder or cooler. This study shows that some vectors had migrated from the central part of Iran with dry and sunny landscape, moved towards the mountainous areas of the north or the warm and humid areas of the south. Historical records show that these anophelines have previously been distributed in lowland areas. If this process continues in the future, Anopheles mosquitoes may be seen in low lands with cold areas in central and northern parts of the country or will occupy humid and warm climates in the southern parts of the country where water is more available.

This study was designed to identify populations of Rhipicephalus sanguineus collected from Iran and also to study molecular taxonomy of Rhipicephalus species using cytochrome C oxidase subunit 1 (COI) and internal transcribed spacer 2 (ITS2) sequences. Tick specimens were collected from livestock (sheep and goat) in 14 Iranian provinces. DNA of individual specimens was extracted and PCR was done on these samples. So, 62 sequencing (33 COI and 29 ITS2) were done, successfully. Morphologically, we identified four Rhipicephalus species, namely R. bursa, R. sanguineus (s.l.), R. sanguineus (s.s.), and R. turanicus based on taxonomic keys. The data obtained from the phylogenetic analyses of COI and ITS2 fragments present a possible conflict regarding the identity of R. sanguineus species. Thus, the molecular identification of R. sanguineus group might be different according to mitochondrial and nuclear DNA. The results show a phylogenic conflict based on COI and ITS2 phylogeny in a tree topology. We dealt with three genetic entities in R. sanguineus group (i.e. R. sanguineus (s.s.), R. sanguineus (s.l.), and R. turanicus) based on COI phylogeny and two genetic clades (i.e. R. sanguineus (s.s.) and R. sanguineus (s.l.)/R. turanicus) according to ITS2 phylogeny.

Background Different mosquito-borne pathogens are circulating in Iran including Sindbis virus, West Nile virus, filarioid worms and malaria parasites. However, the local transmission cycles of these pathogenic agents are poorly understood, especially because ecological data on vector species are scarce and there is limited knowledge about the host range; this understanding could help to direct species-specific vector control measurements or to prioritize research. Methods In the summers of 2015 and 2016, blood-fed mosquitoes were collected at 13 trapping sites on the coast of the Caspian Sea in northern Iran and at an additional trapping site in western Iran. Mosquitoes were generally collected with either a Biogents Sentinel trap or a Heavy Duty Encephalitis Vector Survey trap installed outside. A handheld aspirator was used at the trapping site in western Iran, in addition to a few samplings around the other trapping sites. On average, eight trapping periods were conducted per trapping site. The sources of blood meals were identified using a DNA barcoding approach targeting the cytochrome b or 16S rRNA gene fragment. Results The source of blood meals for 580 blood-fed mosquito specimens of 20 different taxa were determined, resulting in the identification of 13 different host species (9 mammals including humans, 3 birds and 1 reptile), whereby no mixed blood meals were detected. Five mosquito species represented more than 85.8% of all collected blood-fed specimens: Culex pipiens pipiens form pipiens (305 specimens, 55.7% of all mosquito specimens), Cx. theileri (60, 10.9%), Cx. sitiens (51, 9.3%), Cx. perexiguus (29, 5.3%) and Anopheles superpictus (25, 4.6%). The most commonly detected hosts of the four most abundant mosquito species were humans (Homo sapiens; 224 mosquito specimens, 40.9% of all mosquito specimens), cattle (Bos taurus; 171, 31.2%) and ducks (Anas spp.; 75, 13.7%). These four mosquito species had similar host-feeding patterns. The only exceptions were a relatively high proportion of birds for Cx. pipiens pipiens f. pipiens (23.2% of detected blood meal sources) and a high proportion of non-human mammals for Cx. theileri (73.4%). Trapping month, surrounding area, or trapping method had no statistically significant impact on the observed host-feeding patterns of Cx. pipiens pipiens f. pipiens. Conclusions Due to the diverse and overlapping host-feeding patterns, several mosquito species must be considered as potential enzootic and bridge vectors for diverse mosquito-borne pathogens in Iran. Most species can potentially transmit pathogens between mammals as well as between mammals and birds, which might be the result of a similar host selection or a high dependence on the host availability. Electronic supplementary material The online version of this article (10.1186/s13071-018-3237-2) contains supplementary material, which is available to authorized users.

Zoonotic cutaneous leishmaniasis (ZCL), caused by Leishmania major, is a common zoonotic vector-borne disease in Iran. Close contact with infected reservoir hosts increases the probability of transmission of Leishmania parasite infections to susceptible humans. Four gerbil species (Rodentia: Gerbillidae) serve as the main reservoir hosts for ZCL in different endemic foci of Iran. These species include Rhombomys opimus, Meriones libycus, Meriones hurrianae and Tatera indica; while notable infection has been reported in Nesokia indica as well. The purpose of this study is to model the distribution of these reservoirs to identify the risk areas of ZCL. A data bank was developed including all published data during the period of 1970-2015. Maximum entropy model was used to find the most appropriate ecological niches for each species. The areas under curve obtained were 0.961, 0.927, 0.922, 0.997 and 0.899, instead of 1, for training test in R. opimus, M. libycus, T. indica, M. hurrianae and N. indica, respectively. The environmental variable with the highest gain when used in isolation was slope for R. opimus and N. indica, annual mean temperature for M. libycus, and seasonal precipitation for T. indica and M. hurrianae. Summation of presence probabilities for three main species, i.e., R. opimus, M. libycus and T. indica revealed favorable ecological niches in wide areas of 16 provinces. This is the first study to predict the distribution of ZCL reservoir hosts in Iran. Climatology and topography variables had high contributions toward the prediction of potential distribution of the main reservoir species; therefore, as climate changes, the models should be updated periodically with novel data, and the results should be used in disease-monitoring programs.

Identifying mosquito species is a fundamental step in risk assessment and implementation of preventative strategies. Moreover, Culex pipiens is the most widespread mosquito vector in several regions of Iran and is the main vector for transmission of West Nile virus (WNV). Mosquitoes were collected at 14 sites in northern regions of Iran in 2015 and 2016. A subset of mosquito specimens was selected for identification confirmation using a DNA-barcoding technique. Construction of a phylogenetic tree showed clustering of mosquito sequences into three main genera: Aedes, Anopheles and Culex with individuals of a single species clustered closely together, regardless of where and when they were collected. Cx. pipiens complex and Cx. torrentium were identified and differentiated using multiplex real-time PCR targeting the gene locus for acetylcholinesterase 2 (ace2) to discriminate between Cx. pipiens pipiens and Cx. torrentium. The CQ11 microsatellite locus was used for discrimination between Cpp. biotypes. The predominant mosquito species in investigated regions were Cx. pipiens pipiens biotype pipiens, but we also detected Culex pipiens pipiens biotype molestus and hybrids of the two pipiens biotypes, as well as Cx. torrentium. The results of this study represent the first certain evidence of the presence of Cx. pipiens pipiens biotype molestus and hybrids between pipiens and molestus forms, and Cx. torrentium in Iran through a molecular identification approach. This report of a potentially important bridge vector for WNV might have key influence in the risk projections for WNV in Iran.

Background Malaria is an important mosquito-borne disease, transmitted to humans by Anopheles mosquitoes. The aim of this study was to gather all records of three main malaria vectors in Iran during the last decades, and to predict the current distribution and the environmental suitability for these species across the country. Methods All published documents on An. superpictus Grassi (s.l.), An. maculipennis Meigen (s.l.) and An. sacharovi Favre during 1970–2016 in Iran were obtained from different online data bases and academic libraries. A database was created in ArcMap 10.3. Ecology of these species was analyzed and the ecological niches were predicted using MaxEnt model. Results Anopheles superpictus (s.l.) is the most widespread malaria vector in Iran, and exists in both malaria endemic and non-endemic areas. Whereas An. maculipennis (s.l.) is reported from the northern and northwestern parts, Anopheles sacharovi is mostly found in the northwestern Iran, although there are some reports of this species in the western, southwestern and eastern parts. The area under receiver operating characteristic (ROC) curve (AUC) for training and testing data was calculated as 0.869 and 0.828, 0.939 and 0.915, and 0.921 and 0.979, for An. superpictus (s.l.), An. maculipennis (s.l.) and An. sacharovi, respectively. Jackknife test showed the environmental variable with highest gain in the predicting power of the model when used in isolation was annual precipitation for An. superpictus (s.l.) and An. maculipennis (s.l.), and precipitation of the driest quarter for An. sacharovi. Conclusions Despite this range, global warming may increase the potential risk for malaria transmission in some cleared-up areas, where these proven vectors are active. Mapping and prediction of spatial/temporal distribution of these vectors will be beneficial for decision makers to be aware of malaria transmission risk, especially in the western parts of the country. Electronic supplementary material The online version of this article (10.1186/s13071-018-2973-7) contains supplementary material, which is available to authorized users.

Background: Malaria has long been regarded as one of the most important public health issues in Iran. Although the country is now in the elimination phase, some endemic foci of malaria are still present in the southeastern areas of the country. In some endemic foci, there are no data on the malaria vectors. To fill this gap, the present study was designed to provide basic entomological data on malaria vectors in the southeastern areas of Iran. Methods: Adult and larval stages of Anopheles mosquitoes were collected by using different catch methods. Resistance of the main malaria vector in the study area to selected insecticides was evaluated using diagnostic doses advised by the World Health Organization in 2013–2014. Results: A total of 3288 larvae and 1055 adult Anopheles mosquitoes were collected, and identified as: Anopheles stephensi (32.1%), Anopheles culicifacies s.l. (23.4%), Anopheles dthali (23.2%), Anopheles superpictus s.l. (12.7%), and Anopheles fluviatilis s.l. (8.6%). Anopheles stephensi was the most predominant mosquito species collected indoors at the study area, with two peaks of activity in May and November. This species was found to be resistant to DDT 4%, tolerant to malathion 5% and susceptible to other tested insecticides. Conclusion: All the five malaria vectors endemic to the south of Iran were collected and identified in the study area. Our findings on the ecology and resting/feeding habitats of these malaria vectors provide information useful for planning vector control program in this malarious area.

Introduction: Hospital beds, human resources, and medical equipment are the costliest elements in the health system and play an essential role at the time of treatment. In this paper, different phases of the NEDA 2026 project and its methodological approach were presented and its formulation process was analysed using the Kingdon model of policymaking. Methods: Iran Health Roadmap (NEDA 2026) project started in March 2016 and ended in March 2017. The main components of this project were hospital beds, clinical human resources, specialist personnel, capital medical equipment, laboratory facilities, emergency services, and service delivery model. Kingdon model of policymaking was used to evaluate NEDA 2026 development and implementation. In this study, all activities to accomplish each step in the Kingdon model was described. Results: The followings were done to accomplish the goals of each step: collecting experts' viewpoint (problem identification and definition), systematic review of the literature, analysis of previous experiences, stakeholder analysis, economic analysis, and feasibility study (solution appropriateness analysis), three-round Delphi survey (policy survey and scrutinization), and intersectoral and interasectoral agreement (policy legislation). Conclusion: In the provision of an efficient health service, various components affect each other and the desired outcome, so they need to be considered as parts of an integrated system in developing a roadmap for the health system. Thus, this study demonstrated the cooperation process at different levels of Iran's health system to formulate a roadmap to provide the necessary resources for the health sector for the next 10 years and to ensure its feasibility using the Kingdon policy framework.

Zoonotic Cutaneous Leishmaniasis (ZCL) is still a serious health problem in Iran. The objective of the study was to determine the differences in sand fly biodiversity in Shush (plain) and Khorramshahr (littoral) Counties, Khuzestan Province, southwest Iran. Sand flies were collected using sticky paper traps from urban, semi urban, agricultural and natural ecotypes. Alpha and beta diversity were calculated using Shannon–Weiner index and Jaccard's and Sorensen's coefficients, respectively. Synanthropic index was determined for the first time for Phlebotomus papatasi and Phlebotomus alexandri in different land use categories in Iran. Totally 11213 specimens, 68.47% in Shush and 31.53% in Khorramshahr, were collected. Eleven species of sand flies including, 2 of genus Phlebotomus and 9 of genus Sergentomyia were identified. Sergentomyia christophersi was found as a new record. Dominant species were P. papatasi and Sergentomyia sintoni. Shannon–Weiner index, richness and evenness in semi urban area of Shush County were more than other habitats. The analysis of α biodiversity showed that agricultural ecosystem of Khorramshahr County had the highest diversity due to maximal richness and diversity and also relatively high evenness. Comparison of similarity of the sand flies population composition between Shush and Khorramshahr indicated the maximum similarity between the urban area of Shush and the semi urban area of Khorramshahr (Sj = 75% and Ss = 86%). Synanthropic index of P. papatasi and P. alexandri were calculated to be −83.34 and −91.18, respectively in Shush County. Estimated synanthropic indices for P. papatasi and P. alexandri in three habitats (natural, semi urban and urban) of Khorramshahr County were −69.84 and −85.89, in the same order. The factors for having high diversity of sand flies in the plain area studied may be due to higher annual precipitation, the related land use and land cover. The changes on the composition of sand flies are perhaps due to human intervention in their natural habitats.

Objective: To study agents of animal wound myiasis in various geographical districts of Fars province. Methods: This study has been done in Fars province, located in the southern part of Iran. Sums of 10358 domestic animals have been visited from April 2011 to March 2012. The infected wounds in any parts of animal body were sampled by means of forceps. Results: About 61% of all animal wound myiasis were caused by larvae of Wohlfahrtia magnifica. The most wound myiasis cases due to this species occurred in central part of Fars province. There wasn't any significant difference between sheep and goat in infestation with myiasis (P>0.05). The infestation rate of myiasis in cattle community was 0.86%. Conclusions: The infestation rate of livestock was lower than other works in Iran and some other countries like Saudi Arabia. Chrysomya bezziana has been mentioned as main myiasis agent in Iran. But in this study it cleared that similarly to some European countries, the common animal myiasis agent in Iran is Wohlfahrtia magnifica. Introducing new species as principal agent for myiasis can help public health and animal husbandry policy makers to prepare sufficient and effective control and/or preventive measures for this disease.

Recent epidemiological evidences revealed the higher prevalence of 'O' blood group in the residents of malaria-endemic areas. Also some data indicated preference of mosquitoes to 'O' group. The aim of this study was to determine ABO group ratio in the residents as well as ABO group preference of Anopheles in two malaria endemic areas in south of Iran. Agglutination method was used for ABO typing of residents. Field blood fed Anopheles specimens were tested against vertebrate DNA using mtDNA-cytB PCR-RFLP and then the human fed specimens were tested for ABO groups using multiplex allele-specific PCR. A total of 409 human blood samples were identified, of which 150(36.7%) were 'O' group followed by 113(27.6%), 109(26.7%), and 37(9.0%) of A, B, and AB groups respectively. Analyzing of 95 blood fed mosquitoes revealed that only four Anopheles stephensi had fed human blood with A(1), B(1), and AB(2) groups. Result of this study revealed high prevalence of O group in south of Iran. To our knowledge, it is the first ABO molecular typing of blood meal in mosquitoes; however, due to low number of human blood fed specimens, ABO host choice of the mosquitoes remains unknown. This study revealed that ABO blood preference of malaria vectors and other arthropod vectors deserves future research.

To the Editor: Tickborne relapsing fever (TBRF) is endemic in Iran; >1,400 cases were confirmed in 19 provinces during 1997–2006 (1). In the western, northwestern, and foothill regions of the Alborz Mountains, the Argasid soft tick Ornithodoros tholozani is commonplace and accounts for ≈60% of TBRF cases attributed to Borrelia persica. However, in central and western Iran, O. tholozani and B. microti–infected O. erraticus ticks coexist (1,2). Two other Borrelia species, B. latyschewii and B. baltazardi, have also been described in northeastern and northwestern Iran (3,4), but no recent human infections with these species have been documented. Cases of TBRF occurring in southern Iran have presumably been caused by B. microti because its tick vector, O. erraticus, predominates in this region. Relapsing fever infections in Hormozgan Province in southern Iran are commonly identified during routine checks for malaria. During 2011–2013, blood samples were obtained from 14 febrile patients referred to medical centers in Jask and Rodan in Hormozgan Province (Technical Appendix Figure). Informed verbal consent was obtained from all participants, and the ethical committee of Pasteur Institute of Iran approved the project. Patients seeking care had fever and >1 sign or symptom, such as headache, chills, sweating, or fatigue. Six patients reported recurrent fever and generalized muscle and joint pain. Each patient lived in a local tent, called a kapar, or in a brick or concrete-block house. Thick and thin blood smears were prepared from blood samples, stained with Giemsa, and examined. None showed malaria parasites; however, spirochetes were observed in thick or thin smears from 3 patients (Technical Appendix Table). Patients whose samples tested positive by microscopy were treated with 500 mg tetracycline every 6 hours for 10 days and became afebrile. DNA was extracted from patients’ serum samples by using the Miniprep DNA kit (QIAGEN, Hilden, Germany) and screened for borrelia DNA by using real-time PCR; negative and positive control DNA from B. microti or B. persica was also screened. Borrelia spp. DNA was detected in 5 (36%) of 14 serum samples (Technical Appendix Table). Of these 5 samples, 2 were also positive by nested PCR that targeted the intergenic spacer (IGS) region (5). The 2 IGS regions were sequenced (ABI-3130XL sequencer; Applied Biosystems, Foster City, CA, USA) in both directions at the Pasteur Institute of Iran. The resulting 539- and 527-bp IGS sequences (GenBank accession nos. {"type":"entrez-nucleotide","attrs":{"text":"KM271987","term_id":"723219919","term_text":"KM271987"}}KM271987 and {"type":"entrez-nucleotide","attrs":{"text":"KM271988","term_id":"723219920","term_text":"KM271988"}}KM271988, respectively) were 97% homologous with B. recurrentis and B. duttonii from Africa (GenBank accession nos. {"type":"entrez-nucleotide","attrs":{"text":"CP000993","term_id":"201084691","term_text":"CP000993"}}CP000993 and {"type":"entrez-nucleotide","attrs":{"text":"DQ000280","term_id":"66276904","term_text":"DQ000280"}}DQ000280, respectively); 96% homologous with B. microti from Iran (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"JQ436580","term_id":"392856345","term_text":"JQ436580"}}JQ436580); and 92% homologous with B. crocidurae from Africa (GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"GU350723","term_id":"300086694","term_text":"GU350723"}}GU350723). A neighbor-joining phylogenetic tree was constructed by using MEGA6 (http://www.megasoftware.net); the 2 IGS sequences clustered into a distinct group separate from B. microti, B. duttonii, and B. recurrentis genotypes (Figure). Figure Phylogenetic tree of Borrelia spp. strains isolated in Iran, 2014. Constructed on the basis of intergenic spacer sequences, the tree is drawn to scale using evolutionary distance computed using the Jukes-Cantor method in which the units reflect substitutions ... B. microti was expected to be found because O. tholozani ticks that transmit B. persica are not seen in southern Iran, but B. microti–infected O. erraticus ticks have been frequently recovered from rodents’ burrows in the region (6). Current molecular data from TBRF borreliae from Iran are limited to 2 isolates of B. persica and B. microti from O. tholozani and O. erraticus ticks, respectively (5,7,8). In situ IGS analysis revealed that spirochetes in our analysis had highest homology (97%) with relapsing fever agents from eastern Africa, B. duttonii and B. recurrentis, followed by B. microti (96%) from Iran (8). B. microti clustered with 1 strain (B. duttonii; GenBank accession no. {"type":"entrez-nucleotide","attrs":{"text":"GU350721","term_id":"300086692","term_text":"GU350721"}}GU350721) and apart from other B. duttonii IGS strains, suggesting that this strain may not be B. duttonii. The phylogenetic tree separated B. duttonii into 4 clades, 2 of which also contained B. recurrentis, confirming previous observations (9) and providing further support that B. recurrentis represents an ecotype of B. duttonii rather than a species (10). Furthermore, the high level of phylogenetic similarity among borreliae from eastern Africa and Iran indicates that the borreliae in our study might represent ecotype-adapted strains. More sequencing of different genomic markers is required to substantiate or refute this possibility. Lack of GenBank data for the remaining borreliae from Iran, B. latyschewii and B. baltazardi, prevent exclusion of these species. Although relapsing fever spirochetes from southern Iran and those from borreliae in Africa have a close phylogenetic similarity, they have different virulence levels and abilities to infect vector and host species. Consequently, deciphering the evolutionary links for these Borrelia spp. is of paramount importance and might provide valued insights into host–microbe interactions. Our report confirms a novel Borrelia IGS sequence type detected in situ from 2 relapsing fever patients. This species showed greatest homology with the relapsing fever borreliae from Africa, B. recurrentis and B. duttonii, but not with B. microti, which is transmitted by O. erraticus ticks, previously believed to be the only soft tick species in this region. These findings challenge the assumption that TBRF in Iran is attributed to only B. persica or B. microti. Technical Appendix. Study site, characteristics, and clinical and laboratory findings for patients having positive results for tickborne relapsing fever, Jask and Rodan Counties, Iran, 2011–2013 Click here to view.(140K, pdf)

Objective To assess molecular characterization, distribution, seasonal activities of sandfly species and Leishmania parasites infecting them for this zoonotic cutaneous leishmaniasis focus. Methods The collections were carried out in 2009–2011 using CDC traps, Sticky Papers and manual aspirator in and around the villages in Abarkouh district. Individual sandflies were characterized by PCR amplification and sequencing of fragments of their mitochondrial cytochrome b gene. Leishmania parasite infections within sandflies were performed by targeting Cyt b, ITS-rDNA, k-DNA and microsatellite genes. Results The PCR assays detected only Leishmania major (L. major). All infections (30) were found in the abundant and widespread vector Phlebotomus papatasi (P. papatasi). Small numbers of other sandfly species were also screened for infections, but none was found. Sergentomyia sintoni and P. papatasi were the predominant members in all locations of this district and in all habitats throughout the trapping season. Only five other sandfly species were found, namely Phlebotomus ansari, Phlebotomus caucasicus, Phlebotomus sergenti, Sergentomyia dentata and Sergentomyia merviney. Conclusions In the current survey, the only infections detected are of L. major in females of P. papatasi (30 out of 190). The rates of infection of P. papatasi by L. major are not significantly different in compare with other locations in Iran with no diversity of parasite strains. Zoonotic cutaneous leishmaniasis may have emerged only recently in Abarkouh district, and the reason may well be the instability of the transmission cycles there.

To carry out the distribution survey of hard ticks of livestock in Boeen Zahra and Takistan counties of Qazvin province from April 2010 to September 2010.Nearly about 2 638 sheep, 461 goats and 318 cattle of 38 herds in different geographical areas were searched for tick infestation.The species compositions collected from the livestock of Boeen Zahra and Takistan were Haemaphysalis concinna (0.63%), Haemaphysalis sulcata (12.66%), Hyalomma anatolicum (3.80%), Hyalomma asiaticum (3.16%), Hyalomma detritum (5.70%), Hyalomma dromedarii (28.48%), Hyalomma marginatum (13.29%), Hyalomma schulzei (1.89%), Rhipicephalus bursa (3.16%) and Rhipicephalus sanguineus (3.16%), and for Takistan's livestock were Hyalomma dromedarii (9.86%), Hyalomma marginatum (13.29%), Hyalomma schulzei (1.89%) and Rhipicephalus sanguineus (3.16%), respectively. Hard ticks compositions in different topographic areas were different. Hyalomma species had the most prevalence in the areas.The veterinary and public health investigation of the above species should be taken.

Bashagard is an important malaria endemic area in south of Iran. An epidemiological and entomological survey was performed during 2002-2010. The aim of study was to determine malaria situation, species composition of anopheline mosquitoes and susceptibility status of main vectors to insecticides/larvicides. A total of 13,490 malaria cases were recorded, Plasmodium vivax (99.64%), P. falciparum (0.35%) and mix cases (0.01%). The highest and lowest Annual Parasite Incidence (API) were observed in 2007 (145.72/1000) and 2009 (6.29/1000), respectively. Anopheles culicifacies, An. dthali, An. stephensi, An. superpictus, An. fluviatilis, An. moghulensis, An. turkhudi and An. apoci were collected from the area. Two peak activities occur in April and October. The first five species were confirmed as malaria vectors in Iran. No indication of sporozoite in mosquitoes using molecular method was performed. Susceptibility tests using diagnostic dose of insecticides and larvicides showed only resistance of An. stephensi to DDT. Tolerance in An. stephensi to deltamethrin and bendiocarb is reported. The same phenomenon was observed in An. culicifacies to DDT, propoxur and deltamethrin, and in An. dthali to malathion and deltamethrin. Larvae of vectors were susceptible to all larvicides, except for An. stephensi that exhibited tolerance to fenthion. In conclusion it should be emphasized that malaria transmission is a complex process in Bashagard. This event is attributed to five proven vectors with different behaviors which are active in the area. Regarding tolerance of vectors to deltamethrin, resistance management is suggested by using new insecticide with novel mode of action.

Background : Using botanical insecticides as an alternative biocontrol technique for vector control is considered by some scientists. Materials and Methods : Chemical composition of the essential oil was analyzed using gas chromatography-mass spectrometry (GC-MS). In addition, the mosquito larvicidal activity of leaf essential oil of Cupressus arizonica was investigated against fourth instar larvae of laboratory-reared An. stephensi according to the method of the World Health Organization. Results : Of 46 constituents in the oil, limonene (14.44%), umbellulone (13.25%) and α-pinene (11%) were determined as the main constituents. Cupressus arizonica volatile oil showed significant larvicidal activity against An. stephensi with LC 50 and LC 90 values 79.30 ppm and 238.89 ppm respectively. Clear dose-response relationships were established with the highest dose of 160 ppm essential oil with almost 100% mortality. Discussion : The results from this study revealed that C. arizonica essential oil could be considered as a natural larvicide against An. stephensi. However, the field evaluation of the formulation is necessary.

Leishmania infantum is the causative agent of infantile visceral leishmaniasis (IVL) in the Mediterranean Basin and, based on isoenzyme typing of the parasite isolated from dogs; this parasite was considered to predominate in the all foci of IVL in Iran. However, based on PCR detection and sequencing of parasite Cysteine Protease B (CPB), only one out of seven sandfly infections in Phlebotomus perfiliewi transcaucasicus was found to be L. infantum in the current investigation. The six other infections were haplotypes of Leishmania donovani, the causative agent of anthroponotic visceral leishmaniasis (AVL) in West Africa and India. The deduced amino acid of the L. donovani haplotype was found to be novel and the shortest CPB protein reported within the Leishmania spp. Circulation of both L. donovani and L. infantum by P. perfiliewi transcaucasicus, in addition to previous data indicating its ability to circulate L. tropica, suggests that this species, like other vectors of VL, is a permissive vector. Finding L. donovani infecting P. perfiliewi transcaucasicus in the area demands extensive and intensive typing of natural Leishmania infections in epidemiological investigations in Iran and the Mediterranean Basin in general.

Mosquitoes of the Anopheles maculipennis complex were collected in nine provinces of Iran (Esfahan, Fars, Gilan, Golestan, Kohkiluyeh va Boyerahmad, Mazandaran, Tehran, Azarbaijan-e Gharbi and Zanjan) between June 1983 and September 2002. The nuclear rDNA ITS2 sequences of 86 specimens were compared with those of seven species of the complex available in GenBank. Three genetically distinct species of the complex were distinguished: A. maculipennis Meigen, A. sacharovi Favre and a previously unrecognized species. The last species is most similar to, but clearly distinct from, A. martinius Shingarev and A. sacharovi. The taxonomy of A. martinius and A. sacharovi is critically reviewed, and justification is provided for formally recognizing the third species as Anopheles persiensissp.n. The new species is the first culicid to be characterized and named principally on the basis of DNA evidence. Anopheles persiensis was collected only in the northern Caspian Sea littoral provinces of Gilan and Mazandaran, and it seems likely that this species could be responsible for malaria transmission in this region that was previously attributed to A. maculipennis. A species-specific RFLP–PCR assay based on ITS2 sequences was developed to facilitate further studies of the three species in Iran.