Genetic Population Diversity of Aedes caspius in Southern Provinces of Iran.

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

Aedes caspius largement distribué dans le monde est le vecteur du virus du chikungunya, du virus West Nile, du virus Tahyna et de la bactérie Francisella tularensis. Les séquences des gènes mitochondriaux COI et COII sont généralement utilisées pour estimer les relations phylogénétiques entre les différents niveaux taxonomiques de cette espèce. Des moustiques adultes ont été collectés sur appâts humains, à l'aide de pièges lumineux de type « CDC/light-trap ¼ et d'aspirateurs en février/avril, juin et octobre/décembre 2013­2015 dans les provinces au sud de l'Iran. Les moustiques ont ensuite été identifiés d'un point de vue morphologique. Trois mille cinq cent soixante-dix moustiques adultes ont été collectés et identifiés comme appartenant à trois genres, dont cinq espèces de Culex, six espèces d'Aedes et une espèce de Culiseta. Dans cette étude, 1 796 spécimens d'Aedes caspius ont été identifiés dans quatre provinces. Les séquences COI et COII des populations d'Ae. caspius ont permis d'identifier respectivement 12 et 11 haplotypes. Les résultats de cette étude ont démontré qu'il existe une forte variation intraspécifique dans ces populations d'Ae. caspius.

Spatial changes in the distribution of malaria vectors during the past 5 decades in Iran.

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

Molecular characterization of sandflies and Leishmania detection in main vector of zoonotic cutaneous leishmaniasis in Abarkouh district of Yazd province, Iran.

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

Identification of chemical constituents and larvicidal activity of Kelussia odoratissima Mozaffarian essential oil against two mosquito vectors Anopheles stephensi and Culex pipiens (Diptera: Culicidae).

The larvicidal activity of essential oil extracted from an indigenous plant, Kelussia odoratissima Mozaffarian was evaluated against two mosquito species, Anopheles stephensi and Culex pipiens. The chemical composition of the essential oil obtained by hydrodistillation from branch tips and leaf of this plant was determined by GC and GC/MS analysis. Forty-nine constituents were identified in the oil. The main constituents of the oil were Z-ligustilide (77.73%), 2-octen-1-ol acetate (6.27%), E-ligustilide (2.27%) and butylidene phthalide (1.97%). Five different logarithmic concentrations of essential oil were evaluated against the 4th instar larvae of An. Stephensi and Cx. pipiens. The LC(50) and LC(90) values against An. stephensi larvae were 4.88 and 9.60 ppm and for Cx. pipiens were 2.69 and 7.90 ppm, respectively. These properties suggest that K. odoratissima oil has potential source of valuable larvicidal compounds for mosquito larval control. This plant which causes high mortality at lower dose could be considered as a highly active plant. In this paper a guideline suggested for larvicidal activity of plant essential oils.

Larval habitats and biodiversity of anopheline mosquitoes (Diptera: Culicidae) in a malarious area of southern Iran.

BACKGROUND & OBJECTIVES: Malaria is the most important mosquito-borne disease in Iran. It is endemic in south to southeastern part of the country. Knowledge about bio-ecology of vectors will support authorities for appropriate management of the disease. Bashagard district is one of the main endemic areas for malaria in south of Iran. This study was conducted to determine anopheline fauna, diversity and affinity in the area, characterization of larval habitats, and mapping their potential distribution across the district. METHODS: The potential aquatic habitats for Anopheles larvae were extracted from Indian Remote Sensing Satellite (IRS) image and digital elevation model of the area using GIS. Surface water bodies were sampled monthly during 2009­10 for anopheline larvae, while characteristics of their physical environment were recorded and water samples were analyzed. RESULTS: A total of 4511 Anopheles larvae were found during the year with the lowest and highest frequencies in February and April, respectively. Dominant species was Anopheles culicifacies. The Shanon diversity index ranged from 0.570­0.829 at fixed collection sites, while the affinity index was significant among some vector species. CONCLUSION: Riversides and riverbeds were the main breeding places which provided sandy, rocky, and clay beds for different species. The potential breeding places as well as distribution of collected species were mapped. Knowledge about ecology of malaria vectors provides information to health sector for effective control programs.

Spatial analysis and mapping of malaria risk in an endemic area, south of Iran: a GIS based decision making for planning of control.

Bashagard district is one of the important malaria endemic areas in southern Iran. From this region a total of 16,199 indigenous cases have been reported in recent years. The aim of this study was to determine the situation of the disease and provide the risk map for the area. ArcGIS9.2 was used for mapping spatial distribution of malaria incidence. Hot spots were obtained using evidence-based weighting method for transmission risk. Environmental factors including temperature, relative humidity, altitude, slope and distance to rivers were combined by weighted multi criteria evaluation for mapping malaria hazard area at the district level. Similarly, risk map was developed by overlaying weighted hazard, land use/land cover, population density, malaria incidence, development factors and intervention methods. Our results reveal that the disease mainly occurs in north and east of the study area. Consequently the district is divided into three strata. Appropriate interventions are recommended for each stratum based on national malaria policy. Malaria hazard and risk map, stratification based on relevant information and data analyzing provide a useful method preparedness and early warning system for malaria control, although regular updating is required timely.

Entomological and epidemiological attributes for malaria transmission and implementation of vector control in southern Iran.

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

Knowledge, attitudes and practices regarding malaria control in an endemic area of southern Iran.

Iran is in the malaria pre-elimination stage. In this situation community based strategies are important for malaria control. In order to intensify elimination activities we need to have a better understanding of the knowledge, attitudes and practices (KAP) of malaria prevention in the community. We carried out a KAP study in one malaria endemic district in southern Iran in 2009 using a structured questionnaire. Five villages with indigenous malaria cases during the previous year and a high potential for anopheline mosquito breeding were selected. All the households in each village were visited and the head of the household or other responsible adult was interviewed. Data were analyzed using SPSS 11.5. A total of 494 respondents from the five villages participated. More than 33% of people in the study area were living in sheds with poor facilities. The illiteracy level of the studied population was high (44.2%) and significantly affected the knowledge and practices of the respondents about malaria control (p<0.05). Descriptive analysis showed significant differences between sex, job and history of malaria infection of the respondents and their knowledge and practices about malaria control (p<0.05). Knowledge of the respondents about the route of transmission in malaria was good; 72.1% knew mosquitoes were the vector. Most people knew at least one symptom of the disease. The practices of respondents were 59.5% for bednet use and 9.3% for screening windows. The knowledge, attitudes and practices of respondents in this survey about malaria control were good in some cases, but in general needed to be improved with educational programs.

Molecular characterization of the Anopheles maculipennis complex in the Islamic Republic of Iran.

A diagnostic polymerase chain reaction (PCR) assay using species-specific primers and direct sequencing was used to identify members of the Anopheles maculipennis complex in the north-west and central regions of the Islamic Republic of Iran. Specimens were collected from 9 provinces during 2 seasonal activities in 2001-2002, identified morphologically and subjected to PCR assay and direct sequencing. Results showed that only 2 species, An. maculipennis Meigen, and An. sacharovi Faver, were present in the area of study. This was confirmed by the high similarity (99.2%-100%) of their sequences with those available in GenBank. The molecular data and relative distribution of these species in relation to their vectorial capacity and the epidemiology of malaria in the region are discussed.

The Anopheles maculipennis complex (Diptera: Culicidae) in Iran: molecular characterization and recognition of a new species.

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

Molecular characterization of the Anopheles maculipennis complex in the Islamic Republic of Iran

A diagnostic polymerase chain reaction [PCR] assay using species-specific primers and direct sequencing was used to identify members of the Anopheles maculipennis complex in the north-west and central regions of the Islamic Republic of Iran. Specimens were collected from 9 provinces during 2 seasonal activities in 2001-2002, identified morphologically and subjected to PCR assay and direct sequencing. Results showed that only 2 species, An. maculipennis Meigen, and An. sacharovi Faver, were present in the area of study. This was confirmed by the high similarity [99.2%-100%] of their sequences with those available in GenBank. The molecular data and relative distribution of these species in relation to their vectorial capacity and the epidemiology of malaria in the region are discussed