Species Composition and Geographic Distribution of Culicinae Mosquitoes and Their Possible Infection with West Nile Virus in Hormozgan Province, Southern Iran.

Background: West Nile virus (WNV) is a mosquito-borne virus that causes a febrile disease and may cause a fatal neurological illness in humans. We aimed to investigate the geographic distribution of Culicinae mosquitoes and their possible infection with WNV in Hormozgan Province, southern Iran. Methods: Field studies were carried out from June 2017 to May 2019. Different mosquito sampling methods were used monthly to collect mosquitoes from 22 sites. The Real-Time PCR technique was used to detect the virus infection in the mosquitoes. Results: Overall, 6165 mosquitoes were caught. The species were Culex theileri (33.25%), Cx. pipiens (20.45%), Cx. quinquefasciatus (10.51%), Aedes caspius (6.33%), Cx. tritaeniorhynchus (5.82%), Ae. vexans (4.10%), Cx. sinaiticus (3.62%), Cx. antennatus (3.29%), Culiseta longiareolata (2.81%), Cx. perexiguus (2.03%), Cs. subochrea (1.95%), Cx. mimeticus (1.49%), Cx. pusillus (1.38%), Cx. univittatus (1.27%), Cx. modestus (1.14%), and Cx. sitiens (0.57%). The molecular detection of virus infection in mosquitoes found to be negative for WNV. Conclusion: The presence of many species of mosquito vectors and high population traffic increase the risk of disease transmission is very high. Therefore, the way to restrict WNV infection factors is increasing the knowledge for personal protection measures to prevent mosquito bites.

Baseline susceptibility of a wild strain of main vectors of leishmaniasis to WHO-recommended insecticides in southeastern Iran.

BACKGROUND: In Iran, both cutaneous leishmaniases (CL) and visceral leishmaniases (VL) are endemic, recording one of the 10 highest CL prevalence in the world. Parasites are transmitted by the bite of infected Phlebotomus sand fly females. Several sand fly species have been identified as vectors in the studied region of Kerman province. Residual spraying to control adult sand flies, is the only way to decrease the spreading of the diseases but, following control treatment against malaria vectors in endemic areas in Iran, resistance or tolerance to insecticides emerged in some sand fly species. The objective of this study was to survey insecticides susceptibility levels of 3 vector species in wild sand fly populations in different foci of the diseases in Kerman province. Ph. sergenti, and Ph. papatasi respectively vectors of anthroponotic and zoonotic cutaneous leishmaniases and for the first time Ph. alexandri one of the anthroponotic visceral leishmaniases vector were tested against: deltamethrin 0.05%, malathion 5%, dichloro-diphenyl-trichloroethane (DDT) 4%. MATERIALS AND METHODS: In leishmaniases endemic areas species specific sand fly sites were selected in Kerman province, and specimens were collected by manual aspirators at different time intervals during the spring and summer 2019. All the susceptibility tests were performed according to the WHO tube test recommended procedure. RESULTS: Twenty five blood-fed female sand flies from the region's prevalent species were used in each pooled test replicates. All wild specimens died within 60 min of exposure to DDT 4%, malathion 5%, and deltamethrin 0.05%, but the mortality rate for Ph. papatasi exposed to malathion and DDT was 91.6% and 66.6%, respectively. CONCLUSION: According to current study results, Ph. sergenti and Ph. alexandri are highly susceptible to all the evaluated insecticides in the study areas. However, Ph. papatasi was susceptible to deltamethrin (100% mortality), possibly resistant or tolerant to malathion (91.6% mortality), and confirmed to be resistant to DDT (66.6% mortality).

Effect of Meteorological Factors on Hyalomma Species Composition and Their Host Preference, Seasonal Prevalence and Infection Status to Crimean-Congo Haemorrhagic Fever in Iran.

BACKGROUND: The impact of environmental factors and host on Hyalomma spp. community structure and abundance in the main Crimean-Congo haemorrhagic fever (CCHF) foci of Iran is largely unknown. METHODS: Biotic and abiotic factors, including host, temperature, humidity, altitude, Köppen-Geiger climate types, season, and precipitation on Hyalomma spp. community structure and abundances in 11 provinces of Iran were investigated. Additionally, the possible infection of ticks with CCHF virus was evaluated using reverse transcription PCR technique. RESULTS: Species analyses demonstrated the presence of Hyalomma anatolicum, H. marginatum, H. dromedarii, H. asiaticum, H. detritum and H. schulzei in the study area. Hyalomma anatolicum was the dominant species in the southern and northern parts, whereas H. dromedarii was distributed mostly in central parts of the country. The highest tick infestation was recognized in hot season. Spatial variation in tick relative density was observed between habitat types where more ticks were collected in deserts, semi-deserts, and Mediterranean habitats. Except for H. dromedarii, which was more prevalent on camel (P= 0.044), there were no significant variations in the frequencies of other Hyalomma species on different hosts. Hyalomma anatolicum, H. dromedarii frequencies had significant positive and negative association with temperature and precipitation respectively. Also humidity has positive impact on H. asiaticum frequency. CONCLUSION: Data presented here will help improve ecological models to forecast the distribution of Hyalomma spp. ticks, to evaluate the risk of CCHF and other tick-borne diseases, and to design proper vector control measures to suppress Hyalomma populations in Iran.

First Report of Biochemical Mechanisms of Insecticide Resistance in the Field Population of Culex pipiens (Diptera: Culicidae) from Sari, Mazandaran, North of Iran.

BACKGROUND: Culex pipiens play an important role in transmission of infectious diseases. Vector control by chemical pesticides, leads inevitably to resistance development. Understanding the underlying resistance mechanisms can help improve the control programmes and insecticide resistance management. METHODS: The total contents of cytochrome p450s and the activities of glutathione S-transferases, alpha- and beta-esterases and inhibition rates of acetylcholine esterase (by propoxur) were measured in the field population of Cx. pipiens collected from Sari County, North of Iran, in 2016 and the results were compared with those of the laboratory susceptible strain according to the biochemical assay methods of WHO for adult mosquitoes. Independent sample t-test was used to compare the mean values of enzyme activities/contents between filed and laboratory susceptible populations. RESULTS: The enzyme ratio of cytochrome p450s, alpha- and beta-esterases in the field population was 2.07, 3.72 and 1.36 respectively when compared with the results of the laboratory population. Although not statistically significant, the mean GSTs activities in the field population was marginally less than the laboratory population (ER=0.92). Acetylcholinesterase was insensitive to propoxur in 62.82% of the individuals of the tested field population. There was a significant difference (P< 0.05) between all values of the activities/contents of the enzyme in the field population except for GSTs compared with the laboratory susceptible strain. The highest enzyme activity was related to alpha esterase. CONCLUSION: The present study showed a range of metabolic mechanisms, comprising p450s and esterases combined with target site insensitivity of AChE, contributing to organophosphate, carbamate and pyrethroid resistance in the field population of Cx. pipiens.

Baseline Susceptibility of Culiseta longiareolata (Diptera: Culicidae) to Different Imagicides, in Eastern Azerbaijan, Iran.

BACKGROUND: Culiseta longiareolata is an important vector for many human diseases such as brucellosis, avian influenza and West Nile encephalitis. It is likely an intermediate host of avian Plasmodium that can transmit Malta fever. The aim of this study was to determine the susceptibility level of Cs. longiareolata to different classes of imagicides which are recommended by World Health Organization . METHODS: Larval stages of the Cs. longiareolata were collected from their natural habitats in Marand County at East Azerbaijan Province, northwestern of Iran in 2017. Adult susceptibility test were carried out with using impregnated papers to insecticides including DDT 4%, Cyfluthrin 0.15%, Deltamethrin 0.05%, Propoxur 0.1% and Fenitrothion 1% by standard test kits. RESULTS: Results showed that Cs. longiareolata adult is more susceptible to pyrethroid and carbamate insecticides. Among tested insecticides, Cyfluthrin was the most toxic against Cs. longiareolata with LT50 value of 11.53 minutes and Fenitrothion had the least toxic effect (LT50: 63.39 min). CONCLUSIONS: This study provided a guideline for monitoring and evaluation of insecticide susceptibility tests against Cs. longiareolata mosquitoes for further decision making.

Synergists action of piperonyl butoxide and S,S,S-tributyl phosphorotrithioate on toxicity of carbamate insecticides against Blattella germanica.

OBJECTIVE: To determine the synergists action of piperonyl butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) on toxicity of carbamate insecticides against Blattella germanica in Tehran city. METHODS: In the current study, German cockroach strains were collected from several hospitals and dormitories in Tehran. At the beginning, different concentrations of bendiocarb and carbaryl (insecticides belong to carbamate group) were determined by surface contact on a susceptible strain. Then, the level of susceptibility and type of resistance mechanisms in the collected strains from contaminated sites to the aforementioned insecticides were studied by using PBO and DEF synergists with different insecticide ratios to synergist (1:0, 1:1, 1:2, 1:3). RESULTS: The DEF synergist along with bendiocarb and carbaryl completely eliminated the resistance in all strains but PBO did not completely eliminate the resistance in the strains of Mofid, Alvand, Valiasr hospitals and Shariati dormitory. Generally, the impact of DEF was observed in the removing resistance more than PBO. CONCLUSIONS: In most of these strains, resistance to bendiocarb and carbaryl is completely eliminated by DEF, showing a very high role of estraze enzymes in resistance to bendiocarb and carbaryl. But in most strains PBO does not remove the resistance because other mechanisms, such as reduced cuticle penetration and insensitivity to the acetylcholine esterase enzyme, may be involved.

Baseline Susceptibility of Filarial Vector Culex quinquefasciatus (Diptera: Culicidae) to Five Insecticides with Different Modes of Action in Southeast of Iran.

BACKGROUND: Culex quinquefasciatus (Diptera: Culicidae) is an important vector for many human diseases. The aim of this study was to evaluate the susceptibility level of larval and adult stages of Cu. quinquefasciatus to different groups of WHO recommended insecticides for vector control. METHODS: Larval stages of the Culex mosquitoes were collected from their natural habitats in Rafsanjan County at Kerman Province, southeast of Iran in 2016. Insecticide susceptibility status of adult female Cx. quinquefasciatus against DDT (4%), deltamethrin (0.05%), malathion 5%, and bendiocarb (0.1%) were determined using WHO standard insecticide susceptibility test. Additional test was carried out to determine the susceptibility status of larvae of Cx. quinquefasciatus to temephos. Bioassay data were analyzed by Probit program. RESULTS: Cx. quinquefasciatus adults showed resistance to all four groups of the tested insecticides according to the WHO criteria for resistance evaluation. The lethal concentrations for 50% mortality (LC50) and 90% mortality (LC90) of temephos against Cx. quinquefasciatus larvae were 0.18mg/l and 0.78mg/l, respectively. This finding also confirms resistance to temephos based on the WHO recommended instructions for resistance evaluation. CONCLUSION: Resistance to all groups of the tested insecticides should be considered for future vector control investigations in the study area.

High Insecticides Resistance in Culex pipiens (Diptera: Culicidae) from Tehran, Capital of Iran.

BACKGROUND: During recent years transmission of Dirofilaria immitis (dog heart worm) by Culex pipiens and West Nile virus have been reported from Iran. The present study was preformed for evaluating the susceptibility status of Cx. pipiens collected from capital city of Tehran, Iran. METHODS: Four Insecticides including: DDT 4%, Lambdacyhalothrin 0.05%, Deltamethrin 0.05% and Cyfluthrin 0.15 % according to WHO standard methods were used for evaluating the susceptibility status of Cx. pipiens from Tehran moreover For comparison susceptibility status a Laboratory strain also was used. Bioassay data were analyzed using Probit program. The lethal time for 50% and 90% mortality (LT50 and LT90) values were calculated from regression line. RESULTS: The susceptibility status of lab strain of Cx. pipiens revealed that it is susceptible to Lambdacyhalothrin, Deltamethrin, Cyfluthrin and resistant to DDT. Moreover cyfluthrin with LT50=36 seconds and DDT with LT50=3005 seconds had the least and most LT50s. Field population was resistance to all tested insecticides and DDT yielded no mortality. CONCLUSION: Highly resistance level against all WHO recommended imagicides were detected in field populations. We suggest more biochemical and molecular investigations to detect resistance mechanisms in the field population for further decision of vector control.

Current Susceptibility Status of Anopheles stephensi (Diptera: Culicidae) to Different Imagicides in a Malarious Area, Southeastern of Iran.

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.

Detection of Bendiocarb and Carbaryl Resistance Mechanisms among German Cockroach Blattella germanica (Blattaria: Blattellidae) Collected from Tabriz Hospitals, East Azerbaijan Province, Iran in 2013.

BACKGROUND: Insecticide resistance is one of the serious problems for German cockroach control program. This study was conducted to determine the bendiocarb and Carbaryl resistance mechanisms in German cockroaches using the piperonyl butoxide (PBO). METHODS: Bioassay tests were conducted with 4 to 6 different concentrations of both insecticides with four replicates of 10 susceptible strain cockroaches per concentration to determine of discriminative concentration. After determining discriminative concentration, the result was compared to wild strain. The levels of susceptibility and resistance ratio (RR) and synergism ratio (SR) were calculated for each five wild strains. Moreover resistance mechanisms in wild strains were determined using PBO synergist in vivo. RESULTS: Hospital strains showed different levels of resistance to bendiocarb and carbaryl compared to susceptible strain. The bendiocarb and carbaryl resistance ratios ranged from 2.11 to 7.97 and 1.67 to 2 at LD50 levels, respectively. The synergist PBO significantly enhanced the toxicity of bendiocarb and carbaryl to all strains with different degrees of synergist ratio, 1.31, 1.39, 3.61, 1.78, 1.62 and 2.1 fold for bendiocarb, 1.19, 1.18, 1.12. 1.29, 1.45 and 1.11-fold for carbaryl, suggesting monooxygenase involvement in bendiocarb and carbaryl resistance. CONCLUSION: The synergetic effect of PBO had the highest effect on bendiocarb and resistance level was significantly reduced, which indicates the important role of monoxidase enzyme in creating resistance to Bendiocarb. Piperonyl butoxide did not have a significant synergistic effect on carbaryl and did not significantly break the resistance.

Biodiversity of Aquatic Insects of Zayandeh Roud River and Its Branches, Isfahan Province, Iran.

BACKGROUND: Aquatic insects are the major groups of arthropods that spend some parts of their life cycle in the water. These insects play an important role for transmission of some human and animal diseases. There is few information about the aquatic insects fauna of Iran. METHODS: To study the aquatic insects fauna, adult, nymphal and larval collections were carried out from different habitats using the standard technique in Zayandeh Roud River, Isfahan Province,central Iran, during summer 2011. RESULTS: In total, 741 speimens of aquatic insects were collected and morphologically identified. They include 7 families and 12 genera representing 2 Orders. The order of Diptera (92.31%) and Coleoptera (7.69%). The families Culicidae, Syrphidae and Chironomidae from Diptera order, Gyrinidae, Dytiscidae, Haliplidae, Hydrophilidae from Coleoptera order were identified. CONCLUSION: Some aquatic insects play an important role for transmission of human and animal diseases. These insects also are important for biological control. Therefore ecological study on aquatic insects can provide information about ecology of insects in an area for any decision making.

Crimean--Congo hemorrhagic fever: a molecular survey on hard ticks (Ixodidae) in Yazd province, Iran

OBJECTIVE@#To determine the rate of Crimean--Congo hemorrhagic fever virus (CCHFV) infection in hard ticks (Ixodidae) in Yazd province of Iran.@*METHODS@#A molecular survey on hard ticks (Ixodidae) was conducted in Yazd province during 2008-2009. A total of 140 hard ticks (three genera and 7 species) were collected from randomly selected villages and were exanimate for presence of CCHFV reverse transcription--polymerase chain reaction (RT-PCR) method.@*RESULTS@#CCHFV genome was found in 5.71% of hard ticks. All positive ticks were from Hyalomma genus. Positive ticks including: Hyalomma dromedarii, Hyalomma marginatum, Hyalomma anatolicum, Hyalomma detritum, Hyalomma asiaticum. We were not able to find virus in in Rhipicephalus sanguineus and Dermacentor marginatus. Results exhibited that Hyalomma is the main vector in the study area.@*CONCLUSIONS@#Due to the presence of virus in 24 provinces' out of 31, we recommend the use of acaricides and repellent to prevent disease transmission among humans. Greta care should be taken by the people who are working in slaughter houses.