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BACKGROUND: Anopheles stephensi is recognized as the main malaria vector in Iran. In recent years, resistance to several insecticide classes, including organochlorine, pyrethroids, and carbamate compounds, has been reported for this medically important malaria vector. The main objective of the present study was to evaluate the insecticide susceptibility status of An. stephensi collected from the southern part of Iran, and to clarify the mechanism of resistance, using bioassay tests and molecular methods comparing the sequence of susceptible and resistant mosquitoes. METHODS: Mosquito larvae were collected from various larval habitats across six different districts (Gabrik, Sardasht, Tidar, Dehbarez, Kishi and Bandar Abbas) in Hormozgan Provine, located in the southern part of Iran. From each district standing water areas with the highest densities of Anopheles larvae were selected for sampling, and adult mosquitoes were reared from them. Finally, the collected mosquito species were identified using valid keys. Insecticide susceptibility of An. stephensi was tested using permethrin 0.75%, lambdacyhalothrin 0.05%, deltamethrin 0.05%, and DDT 4%, following the World Health Organization (WHO) test procedures for insecticide resistance monitoring. Additionally, knockdown resistance (kdr) mutation in the voltage-gated sodium channel (vgsc) gene was sequenced and analysed among resistant populations to detect possible molecular mechanisms of observed resistance phenotypes. RESULTS: The susceptibility status of An. stephensi revealed that resistance to DDT and permethrin was found in all districts. Furthermore, resistance to all tested insecticides in An. stephensi was detected in Gabrik, Sardasht, Tidar, and Dehbarez. Analysis of knockdown resistance (kdr) mutations at the vgsc did not show evidence for the presence of this mutation in An. stephensi. CONCLUSION: Based on the results of the current study, it appears that in An. stephensi from Hormozgan Province (Iran), other resistance mechanisms such as biochemical resistance due to detoxification enzymes may be involved due to the absence of the kdr mutation or non-target site resistance. Further investigation is warranted in the future to identify the exact resistance mechanisms in this main malaria vector across the country.
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Anopheles , Resistência a Inseticidas , Inseticidas , Mosquitos Vetores , Mutação , Anopheles/genética , Anopheles/efeitos dos fármacos , Animais , Irã (Geográfico) , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Mosquitos Vetores/genética , Mosquitos Vetores/efeitos dos fármacos , Larva/efeitos dos fármacos , Larva/genética , Piretrinas/farmacologia , Permetrina/farmacologia , DDT/farmacologia , Bioensaio , Nitrilas/farmacologia , FemininoRESUMO
BACKGROUND: The Anopheles hyrcanus group is distributed throughout the Oriental and Palaearctic regions and can transmit diseases such as malaria, Japanese encephalitis virus, and filariasis. This investigation marks the inaugural comprehensive study to undertake a phylogenetic analysis of the constituents of this malaria vector group in the northeastern region of Iran, juxtaposed with documented occurrences from different areas within Iran and worldwide. METHODS: Mosquitoes were collected using various methods from nine different locations in Golestan province from April to December 2023. The collected mosquitoes were identified morphologically using valid taxonomic keys. DNA was isolated using the Sambio™ Kit. COI and ITS2 primers were designed using Oligo7 and GeneRunner. PCR and purification were performed with the Qiagen kit. Subsequently, sequencing was carried out at the Mehr Mam GENE Center using an Applied Biosystems 3730XL sequencer. The nucleotide sequences were then analyzed and aligned with GenBank data using BioEdit. Kimura 2-parameter was Utilized for base substitutions. DNA models were selected based on AIC and BIC criteria. Bayesian and Maximum Likelihood trees were constructed, along with a haplotype network. Molecular diversity statistics computed using DnaSP software. RESULTS: In this study, a total of 819 adult mosquitoes were collected. An. hyrcanus was the second most abundant species, predominantly found in Kalaleh and Turkman counties. The sequenced and edited COI and ITS2 sequences were deposited in GenBank under specific accession numbers. Phylogenetic analyses using ML, BI, and NJ methods confirmed a monophyletic lineage for An. hyrcanus with strong support. Molecular analysis of Iranian An. hyrcanus found 11 diverse haplotypes, with the COI gene displaying low diversity. The ITS2 gene revealed two clades - one associating with Iran, Europe, and Asia; the other originating from southwestern Iran. The haplotype network showed two main groups - one from southwest Iran and the other from north Iran. Iran exhibited six distinct haplotypes, while Turkey showcased the highest diversity. CONCLUSIONS: An. hyrcanus in southwestern Iran exhibits a distinct haplogroup, suggesting possible subspecies differentiation. Additional studies are required to validate this phenomenon.
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Anopheles , Complexo IV da Cadeia de Transporte de Elétrons , Mosquitos Vetores , Filogenia , Animais , Irã (Geográfico) , Anopheles/genética , Anopheles/classificação , Complexo IV da Cadeia de Transporte de Elétrons/genética , Mosquitos Vetores/genética , Mosquitos Vetores/classificação , Haplótipos , Variação Genética , Genética Populacional , Análise de Sequência de DNA , DNA Espaçador Ribossômico/genéticaRESUMO
OBJECTIVES: This study aimed to determine feeding behaviors of Phlebotomus sergenti Parrot, in a new focus of Anthroponotic Cutaneous Leishmaniasis in Bam County, southeast Iran. METHODS: Two methods were used to determine the feeding behavior of Phlebotomus sergenti. In the first method, blood-fed sand flies were captured using a mouth aspirator in human and animal dwellings and consequently, blood meal identification was made using Multiplex PCR. The results were used for calculating Host Feeding Index (HFI) and Forage Ratio (FR) parameters. In the second method, human (Homo sapiens), goat (Capra aegagrus), cattle (Bos taurus), chicken (Gallus gallus) and dog (Canis lupus) were used as baits in tent-baited traps to determine the feeding behavior of Phlebotomus sergenti. RESULTS: Multiplex PCR analysis revealed that the most frequent blood in the stomack of sand flies' were from chicken, but the calculation of the FR revealed that this species prefers canine and poultary blood as meal. Human and animal tent-baited traps revealed that most Phlebotomus sergenti were attracted to chicken rather than the other hosts. CONCLUSIONS: Sand flies are attracted to animals for various reasons such as eating blood, mating on their bodies and laying eggs on their feces. Molecular methods are effective and accurate methods to determine the type of host that sandfly fed on, but they do not show host preferences. The results of the molecular analysis, along with the calculation of HFI and FR, can determine the preferred host of sand flies. The current study revealed that dogs, the secondary reservoir of ACL in Iran, is the first preferred host of Phlebotomus sergenti.
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Leishmaniose Cutânea , Phlebotomus , Psychodidae , Humanos , Cães , Animais , Bovinos , Reação em Cadeia da Polimerase Multiplex , Irã (Geográfico) , Comportamento AlimentarRESUMO
Ae. aegypti is an important vector for transmission of some dangerous arboviral diseases, including Dengue Fever. The present study was conducted (from August 2017 to January 2020) to survey the fauna of Culicine mosquitoes, emphasizing the existence of this invasive species in oriental parts of the country located near the Persian Gulf. Different sampling methods were used to collect all life stages of the mosquito. After morphological identification, a molecular study based on Cytochrome Oxidase (COI) gene-specific primers was performed. Then, the COI gene was sequenced via the Sanger method. A total of 4843 adults and 11,873 larvae were collected (8 species of Culex, one species of Culiseta, and 5 species of Aedes). Fifty-five Ae. aegypti specimens (8 adults and 47 larvae) were identified. Based on the biology and ecological requirements of Ae. aegypti, the possibility of the permanent establishment of this species in the tropical climate of the region is very likely. Considering the detection of this invasive vector mosquito species in Iran and the high incidence of some arboviral diseases in the neighboring countries, and continuous movements of the settlers of these areas, potential outbreaks of arboviral diseases can be predicted. Planning and implementing an immediate surveillance and control program of the vector mosquito is vital to prevent the permanent establishment of this invasive vector mosquito species in southern Iran.
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Aedes , Culex , Culicidae , Aedes/genética , Animais , Complexo IV da Cadeia de Transporte de Elétrons/genética , Espécies Introduzidas , Irã (Geográfico) , Larva/genética , Mosquitos Vetores/genéticaRESUMO
BACKGROUND: Aedes aegypti, the primary vector of various human arboviral diseases, is a significant public health threat. Aedes aegypti was detected in Iran in 2018, in Hormozgan province, but comprehensive information regarding its genetic diversity and origin within the country remains scarce. This study aimed to determine the origin and genetic diversity of Ae. aegypti in southern Iran. METHODS: Aedes aegypti mosquitoes were collected from Bandar Abbas City, Hormozgan Province, southern Iran, between May and July 2022. Specimens were morphologically identified. Origin and assess genetic diversity were assessed based on the mitochondrial DNA-encoded cytochrome c oxidase subunit I (mtDNA-COI) gene. RESULTS: BLAST (basic local alignment search tool) analysis confirmed the accuracy of the morphological identification of all specimens as Ae. aegypti, with 100% similarity to GenBank sequences. Calculated variance and haplotype diversity were 0.502 and 0.00157, respectively. Among the 604 examined nucleotide sequences, only a single site was non-synonymous. Total nucleotide diversity and average pairwise nucleotides were determined as 0.00083 and 0.502, respectively. Fu and Li's D test values were not statistically significant. Strobeck's S statistic value was 0.487, and Tajima's D value was 1.53395; both were not statistically significant (P > 0.10). CONCLUSIONS: Phylogenetic analysis revealed two distinct clades with minimal nucleotide differences and low haplotype diversity, suggesting the recent establishment of Ae. Aegypti in the southern region of Iran. The phylogenetic analysis also indicated an association between Ae. aegypti populations and mosquitoes from Saudi Arabia and Pakistan.
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Aedes , Animais , Humanos , Aedes/genética , Filogenia , Variação Genética , Irã (Geográfico) , Mosquitos Vetores/genética , Genética Populacional , DNA Mitocondrial/genética , NucleotídeosRESUMO
BACKGROUND: Mosquitos due to their role in the transmission of different pathogens to humans are considered as an important group in the phylum Arthropoda. According to the WHO and FAO guideline different groups of insecticide applied for controlling pests in both the agricultural and public health sectors. METHODS: All the data published about resistant status of the mosquitoes Anopheles, Culex, Aedes and Culiseta species were searched on PubMed, Elsevier, Web of Science, Magiran and google scholar. The objectives of this study was to review the trend of resistance to insecticides during 2000-2020 in medically important mosquitoes in Iran. The criteria for resistant are followed according to WHO guideline. RESULTS: The Results showed that there are widespread, multiple resistances in the country to different organochlorine, organophosphates, carbamate and pyrethroids insecticides in the mosquitoes. CONCLUSION: The effect of pesticide residues on the environment could be a cause for selection pressure on mosquitos and lead to insecticides resistance to them. Insecticides resistance is main challenge of the vector control program. Also result will provide a guideline for control of the mosquito-borne diseases in the country as well as the world.
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BACKGROUND: Myiasis is a disease caused by infections of tissues and organs of human and vertebrates body by the larvae of real flies of Diptera which feeding on living or dead tissues of host for a period of time. This report aims to present a case of urogenital myiasis caused by the larvae of Psychoda albipennis (Diptera: Psychodidae) for the first time in Iran. METHODS: In this case report, we present a case of a 9-year-old girl with urogenital myiasis caused by P. albipennis. She presented to Sina Hospital with dysuria and claimed that he had observed several black-grayish colored mobile particles in his urine at different times. The patient lived in Miandoab, West Azerbaijan Province, Iran. RESULTS: In the hospital her urine sample, containing 3 larvae was referred to Entomology lab of the Medical Faculty for identification and characterization. According to morphological factors, the larvae were identified to approximate size of 8-10mm long, white to gray color, thorns and pale scales and a siphon at the posterior end of the body. By comparing the larvae with the reported ones from Turkey, diagnosis was confirmed. CONCLUSION: According to our survey, this is the first observation of urogenital myiasis in East Azerbaijan Province, Iran. Our case illustrates urogenital myiasis caused by P. albipennis in Iran. Urogenital myiasis has not been previously reported from Iran as a human disease.
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BACKGROUND: Ticks are vectors of a wide variety of pathogens that can be transmitted to humans, and tick-borne diseases are a significant public health issue worldwide. The present study was carried out on the hard tick infestation of livestock transported to Rafsanjan slaughter house in the southeast of Iran. METHODS: A cross-sectional survey was carried out biweekly from April to September 2016 to determine tick infestation of the meat-producing animals. All the livestock included in our study were thoroughly inspected for the presence of hard ticks on different parts of their bodies. RESULTS: A total of 258 hard ticks were collected from the body of livestock hosts. The ticks that were sampled were classified into two genera and five species: Hyalomma marginatum, Hy. anatolicum, Hy. asiaticum, Hy. dromedarii, and Rhipicephalus sanguineus. Hyalomma dromedarii was the most abundant species in the study area. More than 50 percent of the sampled ticks were collected from the body of camels brought to the slaughter house however molecular analysis showed no Crimean Congo Hemorrhagic Fever (CCHF) virus infection in tick specimens. The Sex ratio of the sampled hard ticks shows that female tick infestation was more common among the study livestock. CONCLUSION: Due to the crucial role of hard ticks in the transmission of different pathogens to humans, additional investigations are necessary to determine the risk of consumption of infested meat-producing animals in the study area.
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BACKGROUND: The attraction of phlebotomine sand flies to plant and animal hosts is due to the produced chemical compounds, affecting the olfactory receptors of the insects. Therefore, novel and effective methods, such as Attractive Toxic Sugar Baits (ATSB) and Attractive Toxic Baits (ATB), are based on the effective materials that attract sand flies toward the host. The present study was designed to identify the attractive materials in plants and animals for using in ATSB and ATB. METHODS: This cross-sectional study was carried out in July 2018 on endemic areas of leishmaniasis in Iran. Different baits, including mango, nectarine, grape, banana, melon and watermelon, defibrinated blood of cattle, sheep, goat and chicken, urine of cattle, sheep, goat and ultimately, simple and complex chemicals, such as CO2, 1-octanol, lactic acid and human sweat were placed inside the traps, and the rate of the sand flies attraction to these materials was studied. Furthermore, data were analyzed using the Kruskal-Wallis test and Mann Whitney U test. RESULTS: There was a significant difference in the sand flies attraction between the traps containing watermelon, urine of cattle, and sheep, and chemicals such as CO2 and human sweat and the control trap (p< 0.05). CONCLUSION: This study showed that watermelon and CO2 are the potential candidates for using in ATSB and ATB, respectively.
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BACKGROUND: The abundance, diversity, distribution and ecology of mosquitoes (Diptera: Culicidae), especially arbovirus vectors are important indices for arthropod-borne diseases control. METHODS: Larvae and adult mosquitoes were collected using the standard methods from different habitats in nine localities of three counties in the East Azerbaijan Province, Northwestern Iran during June to October 2017. In addition, species richness (R), Simpson's diversity index (D), Shannon-Wiener index (H') and evenness (E) as measures of diversity, were calculated. RESULTS: Overall, 1401 mosquito specimens including 1015 adults and 386 larvae were collected in the study area. The properties of geographical larval habitats were recorded. Four genera along with 10 species were collected and identified, including Anopheles hyrcanus, An. maculipennis s.l., An. superpictus s.l., Aedes caspius, Ae. vexans, Culex pipiens, Cx. theileri, Cx. perexiguus, Culiseta longiareolata and Cs. subochrea. Among the three counties, Ahar region presented the highest species richness (R: 1.5) and diversity values (D: 0.79, H': 1.74, E: 0.73). CONCLUSION: This study provides important information on the diversity, distribution and ecology of ten mosquito species in the region. This information leads to a better understanding of mosquito population dynamics in relation to vector control measures.
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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.
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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 ß- 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.
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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.
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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.
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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.