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Background: Anopheles stephensi, a malaria‐transmitting mosquito species, has developed resistance to various insecticides such as DDT, Dieldrin, Malathion, and synthetic pyrethroids. To combat this issue, the World Health Organization (WHO) suggests using Actellic®300CS and Icon®10CS for Indoor Residual Spraying to tackle pyrethroid‐resistant mosquitoes. The aim of this research project was to evaluate the susceptibility of An. stephensi to certain insecticides at the diagnostic concentration + intensity 5x diagnostic concentration (5XDC) assays in Iran and to study the lasting effectiveness of Actellic®300CS and Icon®10CS against this particular malaria vector. Methods: This study assessed the susceptibility of An. stephensi populations in southern Iran to various insecticides, including deltamethrin 0.05%, DDT 4%, malathion 5%, bendiocarb 0.1%, a synergist assay with PBO 4% combined with deltamethrin 0.05%, and an intensity assay using 5x the diagnostic concentration of deltamethrin (0.25%) and bendiocarb 0.5%. Laboratory cone bioassay tests were conducted to determine the residual effectiveness of Actellic®300 and Icon®10CS insecticides on different surfaces commonly found in households, such as cement, mud, plaster, and wood. The tests were carried out following the WHO test kits and standard testing protocols. Results: The An. stephensi populations in Bandar Abbas were found to be susceptible to malathion 5% and deltamethrin 0.25% (5XDC), but exhibited resistance to DDT, standard concentration of deltamethrin, and both standard and intensity concentrations of bendiocarb. In laboratory cone bioassay tests, An. stephensi mortality rates when exposed to Actellic®300CS and Icon®10CS on different surfaces remained consistently more than 80%. Actellic®300CS achieved more than 80% mortality on all substrates for the entire 300‐day post‐spraying period. Conversely, Icon®10CS maintained mortality rates more than 80% on plaster and wood surfaces for 165 days and on mud and cement surfaces for 270 days post‐spraying. Both Actellic®300CS and Icon®10CS demonstrated 100% mortality within 72 h of each test on all surfaces throughout the entire 300‐day post‐spraying period. Conclusion: The study shows the varying levels of resistance of An. stephensi Bandar Abbas population to different insecticides and demonstrates the consistent performance of Actellic®300CS in controlling these mosquitoes on various surfaces. The findings suggest that long‐lasting CS formulations may be more effective for malaria vector control compared to the current options. Further research is needed to validate these findings in field settings and assess the impact of these insecticides on malaria transmission. [ABSTRACT FROM AUTHOR]

Anopheles stephensi is an efficient vector of malaria parasites in Iran. Despite its importance in malaria transmission, there is a scarcity of accurate predictive models of its rates of development at different temperatures. A laboratory colony of An. stephensi, collected from Bandar Abbas County, southern Iran, was established, and all its developmental stages were maintained in temperature-controlled incubators so that the water temperature set at 5, 8, 10, 12.5, 14, 28, 38, 39.5, 42, and 45(±0.2) °C for different treatments until subsequent adult emergence. The Lower and Upper Developmental Temperatures (LDT and UDT) and the growth degree-day (GDD) were calculated for each development stage. A 12-mo population dynamics survey of the larvae and adults of An. stephensi was performed in 3 malaria-endemic villages (Geno, Hormoodar, and Sarkhoon) of Bandar Abbas County, and the obtained data were matched with the constructed GDD model. Based on the field meteorological and dynamics data, the model was verified in the field and used to determine the appropriate date to start spraying. The LDT was determined to be 8.19, 9.74, 8.42, 5.6, 13.57, and 10.03 °C for egg hatching, first, second, and third ecdysis, pupation, and eclosion events, respectively. The UDT was 38 °C for all developmental stages. The thermal requirement for the development of all immature stages of An stephensi was determined to be 187.7 (±56.3) GDD above the LDT. Therefore, the appropriate date to start residual spraying is when the region's GDD reaches 187.7 (±56.3). Given the climatic conditions in Bandar Abbas County, it is expected that the first activity peak of adult An. stephensi would be in March. Field observations showed that An. stephensi activity starts in February and peaks in March. The GDD model can provide a good estimate for peak An. stephensi activity and indicate the optimal deployment time of residual spraying operations against the multiplication and development of malaria parasites inside the vector. [ABSTRACT FROM AUTHOR]

Background: Anopheles stephensi is the main vector of malaria in Iran. This study aimed to determine the susceptibility of An. stephensi from the south of Iran to bendiocarb and to investigate biochemical and molecular resistance mechanisms in this species. Methods: Wild An. stephensi were collected from Hormozgan Province and reared to the adult stage. The susceptibility test was conducted according to the WHO protocols using bendiocarb impregnated papers supplied by WHO. Also, field An. Stephensi specimens were collected from south of Kerman and Sistan and Baluchistan Provinces. To determine the G119S mutation in the acetylcholinesterase (Ace1) gene, PCR-RFLP using AluI restriction enzyme and PCR direct-sequencing were performed for the three field populations and compared with the available GenBank data. Also, biochemical assays were performed to measure alpha and beta esterases, insensitive acetylcholinesterase, and oxidases in the strains. Results: The bioassay tests showed that the An. stephensi field strain was resistant to bendiocarb (mortality rate 89%). Ace1 gene analysis revealed no G119S in the three field populations. Blast search of sequences revealed 98–99% identity with the Ace1 gene from Pakistan and India respectively. Also, the results of biochemical tests revealed the high activity of non-sensitive acetylcholinesterase, alpha and beta-esterase in the resistant strain compared to the susceptible strain. No G119S was detected in this study additionally the enhanced enzyme activity of esterases and acetylcholinesterase suggesting that resistance was metabolic. Conclusion: The use of alternative malaria control methods and the implementation of resistance management strategies are suggested in the study area. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Background: Given the significance of climate change and its substantial effects on mosquitoes’ habitats, this study was aimed to model the spatial distribution of the main malaria vectors in the south east of Iran. Methods: Several scientific databases between 1980 and 2019 were reviewed to find mosquito species and their spatial information in this area. The archived folders in the center for diseases control and prevention were used to exploit essential data on malaria cases and foci. Three representative concentration pathways (RCP2.6, RCP4.5 and RCP8.5) were chosen to stand for three possible climate scenarios. Finally the potential species distribution of Anopheles stephensi and An. culicifacies s.l. in the 2030s and 2050s horizons were estimated by the Maximum Entropy Model. Results: So far, a total of 39 mosquito species belonging to the family Culicidae have been reported from the study area. In 2019, the total malaria cases have increased by 91% compared to 2015, as well as a sharp rise than 2018 (249%). In that year, 91% of cases were imported from other countries, which caused 40% increase in the new potential foci than in 2018. The Jackknife test demonstrated the annual mean temperature and precipitation of the coldest quarter with the greatest impact on the environmental suitability of the mentioned two species. Conclusion: The effect of climate change on the appearance and recurrence of mosquito-borne diseases has been demonstrated in various studies. Collecting further data and conducting investigation on this issue will improve control management, especially for the malaria vectors. [ABSTRACT FROM AUTHOR]

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

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

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. [ABSTRACT FROM AUTHOR]

Background: Malaria is still a health problem in Iran. There are several vector control activities, including Indoor Residual spraying, using insecticide treated nets and larviciding including Temephos. In addition nuisance mosquitos are prevalent in the urban areas. So that evaluation of this species to larvicide will provide a clue for management of vector control activities. Methods: Two mosquito species were used in this study: Anopheles stephensi were collected from Kazeroun and Culex pipiens from Tehran, capital of Iran. All the tests were carried out according to the WHO method. All the test kis was provided by WHO. Results: Results showed a LC50= 0.0523 and LC90=0.3822 mg/l for An. stephensi. The figure for Cx. pipiens was 0.1838 and 0.8505 mg/l respectively. Conclusion: monitoring of insecticide resistance to Temephos should be evaluated regularly for management of vector control. [ABSTRACT FROM AUTHOR]

Background: Anopheles mosquitoes are an important group of arthropods due to their role in transmission of malaria. The present study was conducted for determination of susceptibility status of Anopheles stephensi to different imagicides collected from malarious area in Chabahar city, Iran. Methods: In the present study seven insecticides including: DDT 4%, lambdacyhalothrin 0.05%, deltamethrin 0.05%, permethrin 0.75%, cyfluthrin 0.15% and etofenprox 0.5% were tested based on WHO method. Regression line was plotted for each insecticide using mortality of different exposure times. Bioassay data were analyzed using Probit software and the lethal time for 50% and 90% mortality (LT50 and LT90) values were calculated. Results: The susceptibility levels of field strain of An. stephensi to the discriminative dose of different imagicides were determined 100, 98, 96, 89, 82 and 62% for etofenprox, permethrin, deltamethrin, lambdacyhalothrin, cyfluthrin and DDT, respectively. Our finding indicated that An. stephensi is resistant to DDT, lambdacyhalothrin and cyfluthrin, and susceptible to etofenprox and permethrin and candidate of resistant to deltamethrin based on WHO criteria. Conclusion: Our findings indicated that An. stephensi is resistant to DDT and some pyrethroid insecticides which can be developed due to application of insecticides in health and agriculture. These results can provide a clue for future chemical control program in the study area. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To identify the biological forms, sporozoite rate and molecular characterization of the Anopheles stephensi (An. stephensi) in Hormozgan and Sistan-Baluchistan provinces, the most important malarious areas in Iran. Methods: Wild live An. stephensi samples were collected from different malarious areas in southern Iran. The biological forms were identified based on number of egg-ridges. Molecular characterization of biological forms was verified by analysis of the mitochondrial cytochrome oxidase subunit I and II (mtDNA-COI/COII). The Plasmodium infection was examined in the wild female specimens by species-specific nested–PCR method. Results: Results showed that all three biological forms including mysorensis, intermediate and type are present in the study areas. Molecular investigations revealed no genetic variation between mtDNA COI/COII sequences of the biological forms and no Plasmodium parasites was detected in the collected mosquito samples. Conclusions: Presence of three biological forms with identical sequences showed that the known biological forms belong to a single taxon and the various vectorial capacities reported for these forms are more likely corresponded to other epidemiological factors than to the morphotype of the populations. Lack of malaria parasite infection in An. stephensi, the most important vector of malaria, may be partly due to the success and achievement of ongoing active malaria control program in the region. [Copyright &y& Elsevier]

Background: Malaria still remains a public health problem in Iran. There are different vector control interventions such as insecticide spraying. The present study was carried out to determine the susceptibility status of Anopheles stephensi larvae to temephos as a national plan for monitoring and mapping of insecticide resistance Methods: Eight different localities in two main malarious provinces were determined as field collecting sites. Mosquitoes were collected from the field and reared in an insectray. Susceptibility assays were carried out according to the WHO method. The laboratory reared susceptible Beech-Lab strain was used for comparison. Data were analyzed using Probit analysis to determine LC50 and LC90values. Results: Susceptibility of An. stephensi to temephos indicated that the LC50ranged from 0.0022 mg/l to 0.0141 mg/l. Although all field strains were susceptible to temephos, considerable variations in temephos resistance ratios of field strains were noticed from all the localities studied in comparison with the susceptible strain. A low level of resistance ratio was noticed in An. stephensi populations except for the Chabahar strain (RR= 4.27 fold). All field-collected An. stephensi populations exhibited homogeneity to the larvicide except for Bandar Abbas and Hormoodar village strains (P> 0.05%). Conclusion: Due to intensive use of temephos in the neighboring countries and occurrence of resistant to this insecticide in the main malaria vector in the region, insecticide resistance gene may evolve in the populations of An. stephensi. If temephos be applied as a larvicide it should be used judiciously for resistance management, as rotation strategy. [ABSTRACT FROM AUTHOR]

Abstract: Objective: To investiagte insecticide resistance in target species for better insecticide resistance managemnet in malaria control programs. Methods: The status of insecticide resistance to different imagicides in Anopheles stephensi (An. stephensi) including DDT 4%, lambdacyhalothrin 0.50%, deltamethrin 0.05%, permethrin 0.75%, cyfluthrin 0.15% and etofenprox 0.50% was performed according to WHO standard method. Results: The mortality rate to lambdacyhalothrin, permethrin, cyfluthrin, deltamethrin, etofenprox and DDT was (88.0±3.2), (92.0±2.7), (52.0±5.0), (96.0±2.2), (90.0±3.0) and (41.0±5.7) percent, respectively at diagnostic dose for one hour exposure time followed by 24 h recovery period. Conclusions: These results showed first indication of pyrethroid resistance in An. stephensi in a malarious area, from southern Iran. There is widespread, multiple resistances in the country in An. stephensi to organochlorine and some report of tolerance to organophosphate insecticides and recently to pyrethroids. However, results of this paper will provide a clue for monitoring and mapping of insecticide resistance in the main malaria vector for implementation of any vector control. [Copyright &y& Elsevier]

District of Jiroft is situated in south-east of Iran which is one of the malarious regions. Anopheles stephensi is considered as one of the main malaria vector in this region. Ecology of this species was studied in the area to understand its vector behavior for implementation of effective vector control measures. Different methods like total catch, pit shelter, night bite collection on human and animal, larval dipping methods were used for species identification, seasonal activity, anthropophilic index and egg morphological characteristics. Anthropophilicity index was assessed by ELISA test. Activity of Anopheles species started at the beginning of April, and its peak occurs in late spring. The larvae were found in the river bed with pools, stagnant streams, slow foothill streams, temporary pools, and slowly moving water with and without vegetation, drainage containers of air conditioner and palm irrigation canals. From different methods of adult collection, it was found that spray sheet collection is the appropriate method. ELISA testing of 144 blood meals of females revealed the anthropophilicity of 11.8% indicating host preference on animal, mainly cow. Ridge length and their number on the egg floats confirmed Anopheles stephensi mysorensis form. This study showed that Anopheles stephensi is the main vector of malaria in the region, although some other species may play a role. Our findings could provide a valuable clue for epidemiology and control of malaria in the southeast of Iran. [ABSTRACT FROM AUTHOR]