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Abstract Background Aedes aegypti is the main vector of arboviral diseases worldwide. The species invaded and became established in southern Iran in 2020. Insecticide-based interventions are primarily used for its control. With insecticide resistance widespread, knowledge of resistance mechanisms is vital for informed deployment of insecticidal interventions, but information from Iranian Ae. aegypti is lacking. Methods Fifty-six Ae. aegypti specimens were collected from the port city of Bandar Lengeh in Hormozgan Province in the South of Iran in 2020 and screened for kdr mutations. The most common kdr mutations in Latin America and Asia (V410L, S989P, V1016G/I and F1534C), especially when present in combinations, are highly predictive of DDT and pyrethroid resistance were detected. Phylogenetic analyses based on the diversity of S989P and V1016G/I mutations were undertaken to assess the phylogeography of these kdr mutations. Results Genotyping all four kdr positions of V410L, S989P, V1016G/I and F1534C revealed that only 16 out of the 56 (28.57%) specimens were homozygous wild type for all kdr mutation sites. Six haplotypes including VSVF (0.537), VSVC (0.107), LSVF (0.016), LSIF (0.071), VPGC (0.257) and LPGC (0.011) were detected in this study. For the first time, 11 specimens harbouring the V410L mutation, and 8 samples with V1016I mutation were found. V410L and V1016I were coincided in 8 specimens. Also, six specimens contained 1016G/I double mutation which was not reported before. Conclusions The relatively high frequency of these kdr mutations in Iranian Ae. aegypti indicates a population exhibiting substantial resistance to pyrethroid insecticides, which are used widely in control operations and household formulations. The detection of the 410L/1016I kdr mutant haplotype in Iranian Ae. aegypti suggests possible convergence of invasive populations from West Africa or Latin America. However, as Iran has very limited maritime/air connections with those African countries, a Latin American origin for the invasive Ae. aegypti in Iran is more plausible. Graphical abstract

Abstract Vector-borne diseases emergence, particularly malaria, present a significant public health challenge worldwide. Anophelines are predominant malaria vectors, with varied distribution, and influenced by environment and climate. This study, in Ghana, modelled environmental suitability for Anopheles stephensi, a potential vector that may threaten advances in malaria and vector control. Understanding this vector’s distribution and dynamics ensures effective malaria and vector control programmes implementation. We explored the MaxEnt ecological modelling method to forecast An. stephensi’s potential hotspots and niches. We analysed environmental and climatic variables to predict spatial distribution and ecological niches of An. stephensi with a spatial resolution of approximately 5 km2. Analysing geospatial and species occurrence data, we identified optimal environmental conditions and important factors for its presence. The model’s most important variables guided hotspot prediction across several ecological zones aside from urban and peri-urban regions. Considering the vector’s complex bionomics, these areas provide varying and adaptable conditions for the vector to colonise and establish. This is shown by the AUC = 0.943 prediction accuracy of the model, which is considered excellent. Based on our predictions, this vector species would thrive in the Greater Accra, Ashanti Central, Upper East, Northern, and North East regions. Forecasting its environmental suitability by ecological niche modelling supports proactive surveillance and focused malaria management strategies. Public health officials can act to reduce the risk of malaria transmission by identifying areas where mosquitoes may breed, which will ultimately improve health outcomes and disease control.

Systems for disease vector control should be effective, efficient, and flexible to be able to tackle contemporary challenges and threats in the control and elimination of vector-borne diseases. As a priority activity towards the strengthening of vector control systems, it has been advocated that countries conduct a vector-control needs assessment. A review was carried out of the perceived needs for disease vector control programs among eleven countries and subnational states in South Asia and the Middle East. In each country or state, independent teams conducted vector control needs assessment with engagement of stakeholders. Important weaknesses were described for malaria, dengue and leishmaniases regarding vector surveillance, insecticide susceptibility testing, monitoring and evaluation of operations, entomological capacity and laboratory infrastructure. In addition, community mobilization and intersectoral collaboration showed important gaps. Countries and states expressed concern about insecticide resistance that could reduce the continued effectiveness of interventions, which demands improved monitoring. Moreover, attainment of disease elimination necessitates enhanced vector surveillance. Vector control needs assessment provided a useful planning tool for systematic strengthening of vector control systems. A limitation in conducting the vector control needs assessment was that it is time- and resource-intensive. To increase the feasibility and utility of national assessments, an abridged version of the guidance should focus on operationally relevant topics of the assessment. Similar reviews are needed in other regions with different contextual conditions.

Background: The saliva and salivary glands of ticks possess a wide range of immuno-pharmacologically active mole­cules that effectively modulate the activity of enzymes, antibodies, and amines that have a role in different biological processes. Derived components from saliva and salivary glands of hard ticks Ixodidae have been characterized as poten­tial natural sources for discovering promising anti-cancer drug candidates. Methods: The anti-cancer activity of salivary gland extracts (SGEs) from Hyalomma anatolicum, Hyalomma drome­darii, Hyalomma marginatum, and Hyalomma schulzei was assessed. MTT assays and flow cytometry were done on the HT-29 colorectal cancer cell line to evaluate the anti-viability and proliferative inhibition. Results: Based on the MTT assay results, the SGEs from Hy. dromedarii had the highest and lowest substantial anti-viability effects on the HT-29 cancer cell and human foreskin fibroblast (HFF) normal cell, respectively. The cytometric assessment revealed a significant increase in the apoptosis and necrosis ratio of the HT-29 cancer cells after treatment with Hy. dromedarii SGEs. Conclusion: The results demonstrated that Hy. dromedarii SGEs have significant anti-proliferative, anti-viability, and apoptotic potential. The result of this study suggests that Hy. dromedarii SGEs is an appropriate candidate for further investigations to identify and purify the mechanisms and molecules involved in the anti-cancer activity of the SGEs.

Abstract Mosquitoes are the most important vectors of serious infectious diseases in the world. Dengue, Zika, chikungunya and yellow fever are emerging and re-emerging infectious diseases, associated with the distribution of two key vectors i.e. Aedes aegypti and Aedes albopictus throughout the world including countries neighbouring Iran. Entomological surveillance was planned and performed monthly from May to December during 2014–2020 in selected counties of Mazandaran Province, North of Iran, by ovitrap, larval collection, hand catch and human baited trap. Overall, 4410 Aedes specimens including 2376 larvae (53.9%) and 2034 (46.1%) adults belonging to six species, namely Aedes vexans, Aedes geniculatus, Aedes caspius, Aedes echinus, Aedes pulcritarsis and Aedes flavescence were collected and morphologically identified. Over the seven years of surveillance, Ae. aegypti and Ae. albopictus were not found by any sampling method. Aedes vexans and Ae. geniculatus were the most abundant species, their populations peaked in October and November and was positively correlated with precipitation and relative humidity. Aedes flavescence was a new species record for the province. A flowchart for planning and implementation of invasive mosquito surveillance for provincial health authorities in the country is proposed. These surveillance efforts provide basic and timely information for the health system to act promptly on integrated and intensified surveillance and control programs should Ae. aegypti and Ae. albopictus detected in the province.

Background: Drosophila melanogaster flies are smooth, low upkeep and safe model organisms, they can be effortless­ly used in different fields of life sciences like genomics, biotechnology, genetics, disease model, and Wolbachia-based approaches to fight vectors and the pathogens they transmit. Methods: Fruit fly specimens were collected in 25 districts (14 provinces) of Iran and their morphological recognition was proven by molecular analysis based on sequence homology of mitochondrial COI barcode region. Essential in­formation and specific requirements were provided for laboratory rearing of D. melanogaster. Results: Drosophila melanogaster colonies were found in 23 out of 25 districts. Also, five related species coincident with D. melanogaster were reported in this study including D. ananassae/D. parapallidosa, D. hydei, D. repleta, Zapri­onus indianus (Diptera: Drosophilidae), and Megaselia scalaris (Diptera: Phoridae). The Iranian D. melanogaster mo­lecular signature and their rearing techniques have been described here. The complete life cycle, from (egg to adult), takes approximately 8 days at 25 °C. Some biological points have been presented with highlighting capturing, rearing, culturing, and embryo collection along with primitive recognition and segregation between females and males have been presented. A recipe for culture media and the quantity of various ingredients have been provided. Conclusion: This is the first report on the D. repleta and D. ananassae/D. parapallidosa species for the country. Re­sults of this study provide efficient and effective rearing procedures which are requirement for both small-scale for fa­cilitating entomological research and large-scale use in justifiable vector control management such as disease model or Dengue control.

Objective: To determine the suitable ecological habitats of Aedes (Ae.) aegypti and Ae. albopictus in Iran due to climate change by the 2070s. Methods: All data relating to the spatial distribution of Ae. aegypti and Ae. albopictus worldwide, which indicated the geographical coordinates of the collection sites of these mosquitoes, were extracted from online scientific websites and entered into an Excel file. The effect of climatic and environmental variables on these mosquitoes was evaluated using the MaxEnt model in the current and future climatic conditions in the 2030s, 2050s, and 2070s. Results: The most suitable areas for the establishment of Ae. aegypti are located in the southern and northern coastal areas of Iran, based on the model outputs. The modelling result for suitable ecological niches of Ae. albopictus shows that in the current climatic conditions, the southern half of Iran from east to west, and parts of the northern coasts are prone to the presence of this species. In the future, some regions, such as Gilan and Golestan provinces, will have more potential to exist/establish Ae. albopictus. Also, according to the different climate change scenarios, suitable habitats for this species will gradually change to the northwest and west of the country. The temperature of the wettest season of the year (Bio8) and average annual temperature (Bio1) were the most effective factors in predicting the model for Ae. aegypti and Ae. albopictus, respectively. Conclusions: It is required to focus on entomological studies using different collection methods in the vulnerable areas of Iran. The future modelling results can also be used for long-term planning to prevent the entry and establishment of these invasive Aedes vectors in the country.

Background: Anopheles stephensi is a major vector of malaria in some parts of the world. A standard method for de­termining resistance in adult mosquito populations is the bioassay test recommended by the world health organization (WHO). The papers used in this method have an expiry date. This study aimed to determine the effectiveness of outdat­ed susceptibility test papers for use in insecticide resistance monitoring programs. Methods: Beech and Bandar Abbas strains of An. stephensi were reared in the insectary. Permethrin 0.75%, Deltame­thrin 0.05%, and Bendiocarb 0.1% impregnated test papers prepared by Universiti Sains Malaysia were used. Probit analysis was used to analyze the results and prepare time-mortality regression lines of LT50 and LT90. Results: There was a difference in the mortality of both tested strains of An. stephensi was exposed to all tested insecti­cides. Both expired and not expired Permethrin and Deltamethrin papers induced 100% mortality at the diagnostic time (60min), but their insecticidal properties were reduced gradually in serial times. The highest efficacy of test papers was in the first trimester after the expiry date and decreased over time. Conclusion: At the diagnostic time of 60 minutes, the mortality rate of both dated and expired papers was 100% in the pyrethroid insecticides, even three years after expiry dates, if stored in the package provided by the producer, in a re­frigerator. This value was reduced to less than 100% in the expired papers of Bendiocarb comparing the dated papers that induced 100% mortality.

BackgroundDengue fever is a rapidly emerging infection worldwide with a high public health burden. Adequate training of healthcare workers is essential to warrant the timely provision of health services to improve the outcome of dengue management.MethodsThis is an analytical cross-sectional study, conducted to assess the knowledge, attitudes and practice (KAP) of healthcare workers regarding dengue from April 2021 to March 2022 in Mazandaran Province, northern Iran. Data was collected using a researcher-made structured questionnaire, prepared as Google Forms, and sent to target groups through social media and email. Data analysis was performed by SPSS 22 software using descriptive and inferential statistics (Chi-square) at a significant level of 5%.ResultsMost of the respondents had heard about dengue (83.8%); media (32.7%) and academic education (25%) were the main sources of information. Respondents had less knowledge associated with dengue symptoms (52%) than prevention and control (69%), transmission (72.2%) and clinical management (81%). Based on the 70% cut-off point, the majority of the participants had a good attitude (81%) and practice (73%). However, only 49.6% of the respondents showed good practice regarding dengue local transmission. A significant difference was observed between participants knowledge on clinical management with occupation; attitude with gender and occupation; and practice with gender (p

Phleboviruses are classified into two main groups: the sandfly fever group (transmitted by sandflies and mosquitoes) and the Uukuniemi group (transmitted by ticks). Old World sandfly-borne viruses (SBVs) are classified into four main serocomplexes; sandfly fever Naples viruses (SFNVs), sandfly fever Sicilian viruses (SFSVs), Karimabad viruses (KARVs), and Salehabad viruses (SALVs). This study addresses current knowledge gaps on SBVs in Iran by focusing on identification and molecular epidemiology. We used PCR to examine DNA/RNA extracts to identify sandfly species and evaluate for SBV presence. We identified five specimens positive for phleboviruses: one Ph. sergenti for Tehran virus (TEHV), one Ph. papatasi for SFSV, and two Ph. papatasi and one Ph. perfiliewi for KARV. A phylogenetic tree indicated that the TEHV isolate from this study formed a cluster with previous isolates of TEHV, Zerdali virus, and Fermo virus. Meanwhile, the identified SFSV isolate fell in lineage I and was grouped with previous isolates of SFSVs and Dashli virus in Iran. Finally, the KARV isolates from this study formed a monophyletic clade in a sister relationship with other viruses in KARV lineages I and II. This comprehensive study on SBVs in Iran provided new insights into the molecular epidemiology of TEHV, SFSVs and KARVs in this country.

Background: Dengue fever as a mosquito-borne disease, has rapidly spread due to climate change, globalization, and human behavior. Iran is prone to dengue fever, as its vector recently has been found in the country. This study aimed to assess predictors of dengue preventive practices based on Precaution Adoption Process Model (PAPM) factors in West Azerbaijan province, northwest of Iran. Methods: This cross-sectional study conducted on 405 health professionals of communicable disease sector who were interested in study participation. Data-gathering instrument was an online researcher-made questionnaire consisting of demographic characteristics (11 items), questions based on PAPM, and dengue preventive practices (85 items). Content validity and reliability of the instrument, content validity ratio, content validity index, and Cronbach α were utilized, respectively. Descriptive, analytical, and regression analysis using SPSS and STATA were examined. Results: Regression analysis revealed that awareness of appropriate actions for dengue prevention was a stronger pre­dictor of preventive practice in borderline and appropriate categories (β= 4.09, p< 0.001) and (β= 4.42, p< 0.001), re­spectively. Among factors of PAPM, beliefs about precaution effectiveness and difficulty in borderline (β= 1.04, p= 0.04) and appropriate (β= 1.12, p= 0.03) groups had direct and significant relation with dengue preventive practice. Conclusion: The highest mean score of beliefs about hazard likelihood and severity factor was related to dengue pre­vention. Therefore, theory-based interventions that address beliefs about precaution effectiveness and difficulty can lead to assistance in acting. To improve dengue preventive practices, a well-designed promotive intervention that addresses associated factors in a context-specific manner is essential.

Cutaneous leishmaniasis (CL) is a prevalent infectious disease with considerable morbidity annually. Here, we aimed to investigate the likely variations in gene expression of glycoprotein63 (gp63), heat shock protein 70 (HSP70), histone, arginase, cysteine protease B (CPB), Leishmania homologue of receptors for activated C kinase (LACK), small hydrophilic endoplasmic reticulum-associated protein (SHERP) in metacyclic promastigotes of L. major isolated from Phlebotomus papatasi sand flies and promastigotes excessively cultured in culture medium. The parasites were collected from suspected CL cases in Pasteur Institute of Iran, cultured and inoculated into the female BALB/c mice (2×106 promastigotes). Sand flies were trapped in Qom province, fed with the blood of euthanized infected mice and subsequently dissected in order to isolate the midgut including stomodeal valve. The metacyclic promastigotes were isolated from Ph. papatasi (Pro-Ppap) using peanut agglutinin test (PNA), then continuously cultured in RPMI-1640 medium enriched with fetal bovine serum, penicillin (100 U/ml) and streptomycin (100 mg/ml) to reach stationary phase (Pro-Stat). The gene expression was evaluated in both parasitic stages (Pro-Ppap and Pro-Stat) using qRT-PCR. Out results showed a significant increased gene expression at Pro-Ppap stage for gp63 (P = 0.002), SHERP (P = 0.001) and histone (P = 0.026) genes, in comparison with Pro-Stat stage. Noticeably, significant changes were, also, demonstrated in 10th to 15th passages [gp63 (P = 0.041), arginase (P = 0.016), LACK (P = 0.025)] and in 5th to 20th passage (SHERP) (P = 0.029). In conclusion, the findings of the present study seem to be essential in designing Leishmania studies, in particular regarding host-parasite interaction, immunization and infectivity studies.

Abstract 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.

Background: Fauna and larval habitat characteristics studies on mosquitoes are important tools to identify the breeding places of the vectors and management of the control strategies. This study was done to provide data on Culicidae fauna, larval habitat characteristics and identifying potential vectors of West Nile virus in Lorestan Province, west of Iran. Methods: Culicidae mosquitoes were collected at three counties and nine site stations from Lorestan Province, west of Iran in 2017. Adult mosquitoes were collected using human and animal bite collection methods, New Jersey and CDC light traps and pit shelters by aspirator. Larva were collected by dipping method. RT-PCR technique was employed for detection of the West Nile virus among mosquito’s samples. Results: 4805 mosquitoes were collected from three counties and nine sites in Lorestan Province during June–October 2017, including 4363 adults and 442 larvae. The most abundant species collected from all counties in both adult and larval stages were Culex pipiens (49.10%), Cx. theileri (31.82%), Anopheles maculipennis (11.09%), An. superpictus (2.66%), An. stephensi (2.12%), Cx. perexiguus (1.89%), An. dthali (1.17%) and An. sacharovi (0.15%) respectively. West Nile virus was detected in none of mosquitoes examined. Conclusion: The results of this study revealed that arbovirus vectors such as Cx. pipiens along with Cx. theileri and Cx. perexiguus are well adapted to a broad range of habitats and different climatic conditions in Lorestan Province. That necessitates further routine surveillance of arboviral infections.

Purpose: Excessive use of chemical insecticides has caused environmental pollution and vector resistance. Herbal essential oils with larvicidal properties are good alternatives to synthetic insecticides.mIn this study, larvicide properties of Myrtus communis essential oil and its nanoemulsion against Anopheles stephensi were investigated. Methods: Components of Myrtus communis essential oil were identified by GC–MS. Nanoemulsion of essential oil was made with Tween 80, Span 20, and water. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) determined particle size and morphology of nanoemulsions. The larvicide activity compared with bulk essential oil. Results: A total of 107 M. communis essential oil compounds were discovered. The morphology of a selected nanoemulsion was spherical. LC50 and LC90 of M. communis essential oil were calculated as 26.1 and 46.2 µg/ml, respectively. The larvicide activity of nanoemulsion increased by 40% compared to the bulk essential oil. The nanoemulsion's larvicide activity (100%) lasted up to 3 days, while the essential oil had larvicide properties only for up to 24 h. Conclusions: Myrtus communis essential oil was found to be an effective larvicide and classified as an active larvicide. The residual efficacy of the nanoformulation of M. communis significantly increased compared with the bulk essential oil.

Background: Mosquitoes are responsible for spreading devastating parasites and pathogens causing some important infectious diseases. The present study was done to better understand and update the fauna of Culicidae and to find out the distribution and the type of their larval habitats in Kashan County. Methods: This study was done in four districts of Kashan County (Central, Qamasr, Niasar and Barzok). Mosquito lar­vae were collected from 23 active larval habitats using a standard 350ml capacity mosquito dipper from April to late December 2019. The collected larvae were transferred to containers containing lactophenol, and after two weeks indi­vidually mounted in Berlese's fluid on a microscope slide and identified to species by morphological characters and valid keys. Results: In this study, a total of 9789 larvae were collected from urban and rural areas in Kashan County. The identified genera were Anopheles, Culiseta and Culex. In this study larvae of An. turkhudi, Cx. perexiguus, Cx. mimeticus, Cx. deserticola and Cs. subochrea were collected for the first time from Kashan County. Conclusion: The results of this study indicate the presence and activity of different mosquito species in Kashan County that some of them are vectors of arbovirus and other vector-borne diseases.

Aims: The aim of this study was to monitor mosquito activity with emphasis on Aedes species in Kashan County. Materials and Methods: This investigation was carried out in Kashan County, from April to late September 2019. The location coordinates of the sampling sites were marked and registered with Global Positioning System deviceand mapped using Arc Map 10.5 software. In this study, mosquitos' larvae were collected from 120 standard ovitraps, which had been placed in 15 locations in Kashan, Qamsar, and Niasar. Cyndon dactylon at 50% concentration was used as an oviposition attractant (250 ml/trap). Ovitraps were visited regularly to check for eggs or larvae. The mosquito's larvae were identified using a systematic key. Results: A total of 535 mosquitos' larvae were collected from ovitraps and were identified under two genera Culex and Culiseta. The collected species were Culex pipiens Linnaeus, 1758 (72.4%), Culex theileri Theobald, 1903 (18.3%), Culex torrentium Martini, 1925 (0.2%) and Culiseta longiareolata (Macquart, 1838) (9.1%). This is the first report of present Cx. torrentium Martini, 1925 in Kashan County. Culex pipiens Linnaeus, 1758 was found at 13 locations from 15. In this study, there were 4.46 larvae per ovitrap. Conclusions: Although no samples of Aedes mosquitoes were captured in this preliminary study, further investigations using different methods of mosquito collection should be done to accurately comment on the current fauna of Kashan Aedes.

Objective: To estimate the infection of ticks to Anaplasma, Ehrlichia, Babesia, Theileria, and Brucellaceae using molecular methods in borderline of Iran, Azerbaijan, and Armenia. Methods: Totally, 2 022 ticks were collected from different livestock. Then, species were diagnosed under stereomicroscope according to valid morphological keys. Tick DNA was extracted followed by PCR to detect Anaplasma, Ehrlichia, Theileria, Babesia and Brucellaceae infection in ticks. Results: A total of 498 males [24.62% (95% CI 22.76%-26.57%)], 741 females [36.64% (95% CI 34.54%-38.79%)], 782 nymphs [38.67% (95% CI 36.55%-40.84%)] and 1 larva [0.04% (95% CI 0.00%-0.28%)] were identified. Among identified samples, we found four genera including Hyalomma, Rhipicephalus, Haemaphysalis, and Dermacentor. Molecular assay revealed that the prevalence of ticks to Anaplasma or Ehrlichia, and Brucellaceae was 22.02% (95% CI 16.01%-29.06%) and 15.03% (95% CI 9.43%-22.26%), respectively. Phylogenetic analysis showed that the identified Anaplasma sp. had the most similarity with Anaplasma centrale, Anaplasma platys, Anaplasma camelii, and Anaplasma phagocytophilum, submitted in GenBank. Furthermore, the detected Ehrlichia sp. and Brucellaceae bacterium had the most similarity with Ehrlichia ruminantium and Mycoplana peli, respectively. However, no sign of the presence of Theileria and Babesia spp. was seen in the studied samples. Conclusions: Anaplasmosis, ehrlichiosis and brucellosis should be considered as important health threats in northwestern Iran and consistent monitoring on infection of ticks and livestock should be performed regularly.

Objective: Different studies have been performed on the prevalence of tick-borne pathogens in different areas of Iran; however, as far as our knowledge, there is no regional meta-analysis available for consideration and estimation of tick species infected with different pathogens in Iran. Methods: In this review, among different databases, a total of 95 publications were included, and the infection of different tick species to different tick-borne pathogens was determined; furthermore, presence of pathogens (with 95% confidence intervals) in tick vectors was calculated separately for each province, using Comprehensive Meta-Analysis version 2 (Biostat, USA). Results: Totally, among all 95 studies, 5 673 out of 33 521 investigated ticks were positive according to different detection methods. Overall estimated presence of pathogens in tick vectors in Iran was 8.6% (95% CI 7.0%-10.6%, P

Background and purpose: Insecticide resistance in mosquitoes particularly malaia may lead to reemergence of malaria epidemics. This study aimed to investigate the effect of oral and intramuscular administration of ivermectin on guinea pigs exposed to Anopheles stephensi mosquitoes, the main malaria vector. Materials and methods: The effect of ivermectin was studied in oral and intramuscular administrations (treatment groups), and untreated (control group), on guinea pigs exposed to 750 Anopheles stephensi mosquitoes. The experiments were done at larval, hatching eggs and pregnancy stages in insectary condition at the School of Public Health, Tehran University of Medical Sciences, 2018. Results: After 24-hours of blood feeding, the mortality rates of An. stephensi were 74.19% and 66.4% in animals with intramuscular and oral administration of ivermectin, respectively, which were significantly different with that of the control group (2.80%) (P< 0.001). The rates of infertility in two experimental groups were 63.63% and 45.23%, respectively and in control group it was 19.42% (P< 0.001). Infertility was found to be significantly different between the experimental groups and the control group (P

Background: Extensive use of chemical larvicides to control larvae, has led to resistance in vectors. More efforts have been conducted the use of natural products such as plant essential oils and their new formulations against disease vectors. Nanoformulation techniques are expected to reduce volatility and increase larvicidal efficacy of essential oils. In this study for the first time, a larvicide nanoemulsion from the essential oil of Acroptilon repens was developed and evaluated against Anopheles stephensi larvae under laboratory conditions. Methods: Fresh samples of A. repens plant were collected from Urmia, West Azarbaijan Prov- ince, Iran. A clevenger type apparatus was used for extracting oil. Components of A. repens essential oil (AEO) were identified by gas chromatography–mass spectrometry (GC–MS). All larvicidal bioassay tests were performed according to the method recommended by the World Health Organization under laboratory condition. Particle size and the morphologies of all pre- pared nanoformulations determined by DLS and TEM analysis. Results: A total of 111 compounds were identified in plant. The LC50 and LC90 values of AEO calculated as 7 ppm and 35 ppm respectively. AEO was able to kill 100% of the larvae in 4 days. Conclusion: The nanoemulsion of AEO showed a weak effect on the larvar mortality. It may therefore be suggested that this kind of nanoemulsion is not appropriate for the formulation as a larvicide. It is important to screen native plant natural products, search for new materials and prepare new formulations to develop alternative interventions with a long-lasting impact.

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.

Abstract 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: Crimean-Congo hemorrhagic fever (CCHF) is a tick-borne viral zoonosis with a considerable fatality rate. The virus is mainly transmitted to humans and animals through the bites of infected hard ticks belonging to the genus Hyalomma. This study aimed to determine the prevalence of CCHFV in ticks in Shahr-e Ray city, one of the meat supplier counties in Tehran Province, Iran. Methods: A molecular survey was carried out on collected ticks from sheep in slaughterhouses and husbandries of Shahr-e Ray city, during 2016-17. The ticks were identified using taxonomic characteristics, and the presence of CCHF virus RNA was investigated using an RT-PCR amplifying a 536 bp of the small segment of the viral genome. Results: The results of this study showed that the Hyalomma ticks were the dominant species in Shahr-e Ray slaughterhouses and husbandries. In none of the assayed ticks, RT-PCR detected CCHFV RNA. Conclusion: A considerable tick infestation with members of genus Hyalomma, among sheep in Shahr-e Ray slaughterhouses and husbandries, indicates the vulnerability of this county of tick-borne infections outbreaks such as CCHF. Therefore, applying acaricides and other preventive strategies should be adopted to control the tick population in Shahr-e Ray.

Frequent use of synthetic larvicides has led to the development of resistance in many species of mosquitoes as well as risk of environmental pollution. Recently, encapsulating essential oils (EOs) in surfactants or polymers is being employed as an approach to control the volatility of EOs as green larvicides. In this research, components of tarragon (Artemisia dracunculus) essential oil were identified by GC-MS analysis. Forty-eight components were identified, with 5 major components including estragole (67.623%), cis-Ocimene (8.691%), beta-Ocimene Y (7.577%), Limonene (4.338%) and 3-Methoxy cinnam aldehyde (1.49%). Tarragon EO was encapsulated in chitosan nanocapsules using ionic gelation method and confirmed by FT-IR analysis. Encapsulation efficiency and size of the chitosan nanocapsules were determined 34.91 ± 2% and 203 ± 16 nm, respectively. For the first time, a long-lasting green larvicide was reported which remained active for 10 days, against Anopheles stephensi. Furthermore, cytotoxicity of the nanoformulation was found to be similar to that of temephos on human skin normal cells (HFFF2). This nanoformulation can be a good alternative for synthetic larvicides due to its long-lasting activity, proper effectiveness and also its green constituents.

Meteorological parameters, have been identified as an important factor involved in the transmission of vector-borne diseases. Mosquitoes are extremely sensitive to weather conditions. The aim of this study was investigate the correlation between meteorological parameters and the abundance of mosquitoes in Kashan County. Mosquitoes were collected using four different traps, including hand catch, animal baited bed net trap (usually a cow), human baited bed net trap and BG-Sentinel trap with CO2 from May to December 2019. A total number of mosquitoes collected were 1756 out of which 1621 (92.31%) were Culex, 22 (1.25%) Culiseta and 113 (6.44%) Anopheles in nine species. Most mosquitoes were collected by BG-Sentinel trap with CO2 (63.78%). Monthly distribution of the mosquitoes indicated different monthly peaks. Their high density were recorded in September and were low in December. The spearman's correlation of the mosquito abundance and the meteorological parameters shows that correlation of the number of total collected mosquitoes with relative humidity and precipitation (Rainfall) was weak negative, and there was week correlation with wind speed, and positive strong correlation with temperature. Data collected with various trap types and mosquito correlation with meteorological parameters in this study can be used for mosquito surveillance and control programs. However, meteorological parameters affect the abundance of mosquitoes, but their impact is complex and most of these variables are species specific.

Background: Mosquito species are highly considering as disease transmission as well as nuisance insects. One of the principal strategy to protect human from the mosquito bites is repellent agents. This study aimed to assess repellency of two organic essential oils, Eucalyptus globulus and Syzygium aromaticum from bites of malaria vector, Anopheles stephensi. Methods: The study was conducted in 2019-2020. The components of essential oils of E. globulus and S. aromaticum was determined using gas chromatography/mass spectrometry. The unfed female mosquitoes aged 2-5 d old were used in all experiments. In vivo Klun and Debboun module bioassays were utilized on human-volunteer skin. The essential oils at serial concentrations were used to find repellent efficacy against Anopheles landings and bites. To find the synergistic effect, four combinations of the essential oils were tested. Results: The main composition of E. globulus essential oil was 1,8-Cineol (78.20%), whereas that of S. aromaticum essential oil was 2-methoxy-3-(2-propenyl) (77.04%). Based on minimum effective dose (≤1% biting), 10% (v/v) of E. globulus showed high landing repellency (77.78%), whereas minimum effective dose of S. aromaticum at concentration of 1% had high landing repellency (88.89%). Among four combinations, the ratio of 1:1 of E. globulus (10%):S. aromaticum (1%) showed the most landing repellency (94.44%). Conclusion: The combinations of two essential oils had the most potential repellency effect against landing of mosquitoes. As essential oils are eco-friendly with less irritation for human skin, E. globulus and S. aromaticum essential oils are recommended as effective and safe mosquito repellents.

Objective: To evaluate the susceptibility of Anopheles stephensi (An. stephensi) Liston, the main malaria vector in southern Iran, to WHO recommended insecticides. Methods: Larvae of An. stephensi were collected from three different larval habitats in both urban and rural area of Bandar Abbas city and one rural area in Rudan county southern Iran. WHO standard method was used for evaluation of adult and larval mosquito susceptibility. Bendiocarb, permethrin, lambda-cyhalothrin, deltamethrin as insecticide and temephos and chlorpyriphos as larvicide were used at the diagnostic dosages recommended by WHO. Results: Findings of this study showed all larval populations of An. stephensi were completely susceptible to temephos and candidate for resistance to chlorpyriphos. Adult mosquitoes in rural areas of Bandar Abbas city were resistant to pyrethroid and carbamate insecticides. Conclusion: Comparison of the results of this survey with previous studies indicates that the resistance to pyrethroids and carbamates in this malaria endemic region is increasing. Wide use of pesticides in agriculture is certainly effective in increasing resistance. The inter-sectoral coordination and collaboration in health and agriculture seem to be necessary to manage insecticide resistance in malaria vectors.

Abstract 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.

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.

Background: Southeastern Iran has been established as an area with the potential to harbor Asian tiger mosquito popu­lations. In 2013, a few numbers of Aedes albopictus were detected in three sampling sites of this region. This field study was aimed to evaluate the efficacy of various traps on monitoring mosquitoes and status of this dengue vector, in five urban and 15 suburban/rural areas. Methods: For this purpose, four adult mosquito traps (BG-sentinel 2, bednet, Malaise, and resting box trap) were used and their efficacy compared. In addition, large numbers of CDC ovitraps were employed, within 12 months. Results: A total of 4878 adult samples including 22 species covering five genera were collected and identified from traps. It was not revealed any collection of Ae. albopictus. Statistical analysis showed no significant difference in mete­orological variables between the two periods, the previous report and the current study. There were significant differ­ences in the total number of mosquitoes collected by various traps in the region across different months. Conclusion: The resulting data collected here on the efficiency of the various trap types can be useful for monitoring the densities of mosquito populations, which is an important component of a vector surveillance system. While the pres­ence of Ae. albopictus was determined in this potential risk area, there is no evidence for its establishment and further monitoring needs to be carried out.

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.

Aim: The present study was carried out to identify the Ixodidae ticks fauna of livestock and their seasonal activity in the cities of Boyer-Ahmad and Dena of Kohgiluyeh Province, south-west of Iran. Materials and Methods: Hard ticks from sheep, goats, and cattle were collected manually, stored in 70% ethanol, and identified using morphological characters. Results: During the study, a total of 1273 hard ticks from four genera, including Rhipicephalus, Hyalomma, Dermacentor, and Haemaphysalis, were collected. Rhipicephalus sanguineus (s.l.) had the highest frequencies in both cities with 62.08 and 62.88% of collected specimens, followed by Hyalomma scupense with 14.36 and 13.54% in Boyer-Ahmad and Dena, respectively. Furthermore, Hyalomma marginatum with only one sample or 0.12% of collected ticks showed the lowest frequencies in the studied areas. Dermacentor marginatus with three samples or 0.37% was recorded only in Boyer-Ahmad, and Haemaphysalis sulcata with two samples or 0.43% was recorded only in Dena. In both cities, sheep were the most infested ruminant, and the ears in sheep and goats were the most affected areas. The highest activity was observed in spring, and the lowest activity was observed in winter and autumn. Conclusion: The results of the present study showed that Hyalomma and Rhipicephalus genera were the most widespread genera in the study areas. Regarding the importance of genera, such as Rhipicephalus, Hyalomma, and Haemaphysalis, in transmitting disease agents and the location of Kohgiloyeh and Boyer-Ahmad Province in the routes of migrant birds, further studies are required to elucidate their exact roles in human and livestock health in these areas.

Abstract 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.

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.

Anopheles maculipennis complex comprises some important malaria vectors in Iran, Middle East, and Europ. The principal way to control of malaria remains on the use of chemical insecticides against its vectors because there is no vaccine for malaria prevention. Extensive use of organophosphate compounds has caused to emergence and distribution of insecticide resistance in Anopheles species in Asia. The current study aimed to the detection of three well-known amino acid substitutions (I114T, L119F, and F120L) in the Glutathione S-Transferases epsilon 2 (GSTe2) gene are associated with DDT and organophosphate insecticides resistance in an Anopheles maculipennis population collected from Iran. Adult samples of An. maculipennis were collected by hand and Total catch in Animal and Human Shelters from Azerbaijan-Gharbi and Zanjan provinces. Following morphological identification, DNA was extracted by YTA Genomic DNA Extraction Mini Kit for amplification of rDNA-ITS2 and GSTe2 fragments. ∼500 bp fragment was amplified using F rDNA-ITS2 and GSTe2 primers. rDNA-ITS2 sequence analysis showed 100% similarity with An. maculipennis. GSTe2 nucleotide sequence similarity within species was 99–100%, while, it was 95–96 % when compared with Anopheles sacharovi GSTe2 sequences available in GenBank. Amino acid sequence comparisons showed a novel amino acid substitution in N148D position with 15.79% frequency. The current study reports new GSTe2 amino acid substitution in An. maculipennis s.s., for the first time. The function of the mutation N148D and its association with resistance phenotype need to validate. However, the integration of these data into the malaria control program still remains a challenge.

[This corrects the article DOI: 10.1371/journal.pone.0207308.].

Background: In this study, larvicidal activity of clove essential oil (EO), as a green and relatively potent larvicide, was compared with its main constituent, Eugenol, against Anopheles stephensi. Methods: High-performance liquid chromatography (HPLC) was used to determine the amount of eugenol, major constituent of clove EO. In addition, larvicidal activity of clove EO and eugenol was evaluated against An. stephensi. Results: The amount of eugenol in clove EO was determined as 67% using HPLC analysis. LC50 and LC90 of clove EO (57.49 and 93.14ppm, respectively) were significantly lower than those of eugenol (86.96 and 128.18 ppm, re­spectively). Conclusion: EO showed more effective than its major component. Considering the lower cost of the essential oil and lower risk in occurrence of resistance in larvae, use of clove EO is preferred as larvicide in comparison with eugenol, against An. stephensi.

Background: Malaria is an important mosquito-borne disease considered as one of the public health concerns across many countries. Anopheles mosquitoes are the main vectors of Plasmodium parasites, which cause malaria. Some of these vectors such as Anopheles maculipennis s.l. and Anopheles sacharovi are considered as complex of sibling species distributed in north of Iran. Methods: This study was conducted in north and northwest of Iran including East Azerbaijan, West Azerbaijan, Ar­dabil, Golestan and North Khorasan provinces with emphasis on the northern borders of the country during 2015–2016. Adult specimens were collected and subjected to morphological identification as well as molecular analysis. Results: Overall, 10405 mosquitoes were collected comprising 21 species. Culex pipiens and Cx. theileri were found as the most frequent species in whole study area. Morphological identification showed that out of 1455 female Anoph­eles specimens, 77% belonged to An. maculipennis Group. Out of the identified species, ITS2 region and COI gene sequences of 8 An. maculipennis s.s. and 31 An. sacharovi representing all provinces were obtained and submitted to GenBank. The COI sequences for An. sacharovi revealed the presence of 9 haplotypes with similarity of 98.17–100%. Conclusion: Some investigations have reported An. martinius as a member of sibling species of An. sacharovi among Iranian Anopheles genus; while based on our study, there was no evidence of the presence of this species in north and northwest of Iran.

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.

Ticks harbor many pathogenic, as well as endosymbiotic and non-pathogenic agents. They are host of a variety of as yet unidentified microbes that continue to be described. In the present study, a Brucella-like bacterium was detected in a Boophilus tick by PCR amplification of a partial fragment of 16S rRNA locus followed sequencing. Our results show that the members of the genus Boophilus may act as vectors of brucellosis in nature, but further studies are required to confirm the real role of ticks as vector or reservoirs of specific Brucella species.

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-3rd or young-4th instar larvae of Anopheles stephensi and Culex pipiens under laboratory condition. The mortality counts were made after 24h and LC50 and LC90 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 LC50 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.

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.

Introduction: Blended learning, a new approach in educational planning, is defined as an applying more than one method, strategy, technique or media in education. Todays, due to the development of infrastructure of Internet networks and the access of most of the students, the Internet can be utilized along with traditional and conventional methods of training. The aim of this study was to compare the students’ learning and satisfaction in combination of lecture and e-learning with conventional lecture methods. Methods: This quasi-experimental study is conducted among the sophomore students of Public Health School, Tehran University of Medical Science in 2012- 2013. Four classes of the school are randomly selected and are divided into two groups. Education in two classes (45 students) was in the form of lecture method and in the other two classes (48 students) was blended method with e-Learning and lecture methods. The students’ knowledge about tuberculosis in two groups was collected and measured by using pre and post-test. This step has been done by sending self-reported electronic questionnaires to the students' email addresses through Google Document software. At the end of educational programs, students' satisfaction and comments about two methods were also collected by questionnaires. Statistical tests such as descriptive methods, paired t-test, independent t-test and ANOVA were done through the SPSS 14 software, and p≤0.05 was considered as significant difference. Results: The mean scores of the lecture and blended groups were 13.18±1.37 and 13.35±1.36, respectively; the difference between the pre-test scores of the two groups was not statistically significant (p=0.535). Knowledge scores increased in both groups after training, and the mean and standard deviation of knowledge scores of the lectures and combined groups were 16.51±0.69 and 16.18±1.06, respectively. The difference between the post-test scores of the two groups was not statistically significant (p=0.112). Students’ satisfaction in blended learning method was higher than lecture method. Conclusion: The results revealed that the blended method is effective in increasing the students' learning rate. E-learning can be used to teach some courses and might be considered as economic aspects. Since in universities of medical sciences in the country, the majority of students have access to the Internet and email address, using e-learning could be used as a supplement to traditional teaching methods or sometimes as educational alternative method because this method of teaching increases the students’ knowledge, satisfaction and attention.

Introduction Leishmania major is the causative agent of zoonotic cutaneous leishmaniasis (ZCL), and great gerbils are the main reservoir hosts in Iran. Abarkouh in central Iran is an emerging focal point for which the reservoir hosts of ZCL are unclear. This research project was designed to detect any Leishmania parasites in different wild rodent species. Methods All rodents captured in 2011 and 2012 from Abarkouh district were identified based on morphological characteristics and by amplification of the rodent cytochrome b (Cyt b) gene. To detect Leishmania infection in rodents, deoxyribonucleic acid (DNA) of each ear was extracted. Internal transcribed spacer-ribosomal deoxyribonucleic acid (ITS-rDNA), microsatellites, kinetoplast deoxyribonucleic acid (kDNA) and cytochrome b genes of Leishmania parasites were amplified by polymerase chain reaction (PCR). Restriction fragment length polymorphism (RFLP) and sequencing were employed to confirm the Leishmania identification. Results Of 68 captured rodents in the region, 55 Rhombomys opimus were identified and nine Leishmania infections (9/55) were found. In addition, eight Meriones libycus and two Tatera indica were sampled, and one of each was confirmed to be infected. Two Meriones persicus and one Mus musculus were sampled with no infection. Conclusions The results showed that all 11 unambiguously positive Leishmania infections were Leishmania major. Only one haplotype of L. major (GenBank access No. EF413075) was found and at least three rodents R. opimus, M. libycus and T. indica—appear to be the main and potential reservoir hosts in this ZCL focus. The reservoir hosts are variable and versatile in small ZCL focal locations.

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.

Background: Recently, essential oils and extracts derived from plants have received much interest as potential bioactive agents against mosquito vectors. Methods: The essential oils extract from fresh peel of ripe fruit of Citrus aurantium and Citrus paradisi were tested against mosquito vector Anopheles stephensi (Diptera: Culicidae) under laboratory condition. Then chemical com­position of the essential oil of C. aurantium was analyzed using gas chromatography-mass spectrometry (GC–MS). Results: The essential oils obtained from C. aurantium, and C. paradisi showed good larviciding effect against An. stephensi with LC50 values 31.20 ppm and 35.71 ppm respectively. Clear dose response relationships were established with the highest dose of 80 ppm plant extract evoking almost 100% mortality. Twenty-one (98.62%) constituents in the leaf oil were identified. The main constituent of the leaf oil was Dl-limonene (94.81). Conclusion: The results obtained from this study suggest that the limonene of peel essential oil of C. aurantium is promising as larvicide against An. stephensi larvae and could be useful in the search for new natural larvicidal compounds.

Malaria is considered a major health problem in Iran. There are different methods for vector control. In this study we tested the larvicidal effects of some Iranian plants. The methanolic extracts of 11 plants were prepared with percolation method. The larvicidal activities of them against malaria vector, Anopheles stephensi were studied using World Health Organization standard method. All LC50 values of methanolic extracts of plants that we screened were lower than 300 ppm. The methanolic extract of aerial parts of Lawsonia inermis and Stachys byzantina showed high larvicidal activity with LC50 values 69.40 ppm and 103.28 ppm respectively. The results obtained from this study suggest that the methanolic extracts of these plants have larvicidal effects against Anopheles stephensi larvae and could be useful in the search for new natural larvicidal compounds.

Background: Crimean-Congo Hemorrhagic Fever (CCHF) is a feverous and hemorrhagic disease endemic in some parts of Iran and caused by an arbovirus related to Bunyaviridae family and Nairovirusgenus. The main virus reser­voir in the nature is ticks, however small vertebrates and a wide range of domestic and wild animals are regarded as reservoir hosts. This study was conducted to determine the infection rate of CCHF virus in hard ticks of Sarpole-Zahab County, Kermanshah province, west of Iran. Methods: From total number of 851 collected ticks from 8 villages, 131 ticks were selected randomlyand investi­gated for detection of CCHF virus using RT-PCR. Results: The virus was found in 3.8% of the tested ticks. Hyalommaanatolicum, H.asiaticum and Rhipicephalus sanguineus species were found to have viral infection, with the highest infection rate (11.11%) in Rh. sanguineus. Conclusion: These findings provide epidemiological evidence for planning control strategies of the disease in the study area.