J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 62 http://jad.tums.ac.ir Published Online: April 27, 2019 Original Article Biodiversity of Mosquitoes (Diptera: Culicidae) with Emphasis on Potential Arbovirus Vectors in East Azerbaijan Province, Northwestern Iran Azim Paksa1; Mohammad Mahdi Sedaghat1; Hassan Vatandoost1,2; Mohammad Reza Yaghoobi-Ershadi1; Seyed Hassan Moosa-Kazemi1; Teimour Hazratian3; Alireza Sanei-Dehkordi4,5; *Mohammad Ali Oshaghi1 1Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2Institute for Environmental Research, Tehran University of Medical Sciences, Tehran, Iran 3Departmemt of Parasitology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran 4Department of Medical Entomology and Vector Control, Faculty of Health, Hormozgan University of Medical Sciences, Bandar Abbas, Iran 5Infectious and Tropical Diseases Research Center, Hormozgan Health Institute, Hormozgan University of Medical Sciences, Bandar Abbas, Iran (Received 7 Mar 2018; accepted 2 Jan 2019) Abstract Background: The abundance, diversity, distribution and ecology of mosquitoes (Diptera: Culicidae), especially ar- bovirus 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 addi- tion, species richness (R), Simpson’s diversity index (D), Shannon–Wiener index (H̕) 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, Cu- lex 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). Conclusions: 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. Keywords: Diversity; Ecology; Mosquitoes; Arbovirus vectors; Iran Introduction Mosquitoes (Diptera: Culicidae) are sub- order of Nematocera and Culicidae family, which are the medically-important species for malaria transmission, types of filariasis, types of encephalitis and other arboviral diseases (1-2). The Culicidae family includes 2 subfamilies, 11 tribes; The Culicidae family includes 2 subfam- ilies, 41 Families, 11 tribes, 113 genera and 3563 species (1-4). The most important genera of the Culicidae family are Anopheles, Culex and Aedes. Among the Culicidae, the Aedes species have the most various species and are extremely important in medical cases (5). Mosquitoes play a crucial role in trans- mission of some important diseases such as malaria, filariasis, dengue fever, yellow fever, chikungunya, West Nile virus and Zika virus which are today among the greatest health problems in the world (6-9). Mosquitoes larvae have the power to col- *Corresponding author: Dr Mohammad Ali Oshaghi, E-mail: moshaghi@sina.tums.ac.ir J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 63 http://jad.tums.ac.ir Published Online: April 27, 2019 onize and live in a wide variety of natural and artificial habitats, such as temporary and per- manent water resources, unclean and clean water, large or small water resources, stagnant or stream waters and even the smallest places where water is gathered, such as buckets of water, pots, tires, bromeliads, animal feet prints and plant leaf axes (10-11). Adult mosquitoes are very diverse bionomically, for example, they are too diverse in host searching, biting behaviors, dispersal and reproductive strate- gies (9). Two factors (abiotic and biotic) affect mosquito life cycle. The biotic factors contain adult sugar and blood meal types, species com- munications, interactions and natural enemies. Physicochemical attributes of larval habitats are abiotic factors, which include the type and contents of water, temperature and rainfall. There are complex interactions between these factors that significantly affect mosquito eco- logical adaptability and vectorial capacity for disease transmission with significant concepts for vector management and control at the lo- cal and regional levels (12-13). Accordingly, investigating biotic and abiotic factors for var- ious mosquito fauna make it easier to moni- tor potential modifications of larval habitats affected by rains, global climate change and man-made activities (12-13). Aedes caspius likely transmit the human pathogen, such as Spiroplasma sabaudiensc and Crisiulospora aedis (14). These mosqui- toes transmit Rift Valley fever virus (RVF), West Nile virus (WNV) and Tahyan virus (15). In the region of Palearctic, Ae. caspius inhab- its in lakes, pools, shores of Great Britain, and fresh-water and lower salt marshes in the con- tinental parts of Europe, Russia, Mongolia, northern China, Pakistan, northeastern Africa, Asia Minor, and the Persian Gulf (17-18). Aedes vexans has several subspecies, the typical species is Ae. vexans vexans, and the subspecies Ae. nipponii vexans in East Asia (19) and Ae. vexans arabiensis in Saudi Ara- bia, Yemen and Africa (3). This species is ex- tremely important in medicine and has more than 30 different viral diseases, including East- ern horse encephalitis, Japanese encephalitis (20), California encephalitis, Western horse en- cephalitis (21), as well as other pathogens such as Tularemia and Dirofilaria immitis (22-23). Aedes vexans along with Ae. aegypti and Cx. quinquefasciatus, have the highest global dis- tribution among other mosquitoes in the world (24-25). Several studies were conducted on com- position, distribution and ecology of mosqui- toes in Iran (26-44). Considering the fact that in the East Azarbaijan Province, some vector mosquitoes of diseases such as West Nile, Diro- filariasis and malaria present, study on the com- position, distribution and ecology of mosquito species is very important in different aspects of vector control programs. This study will add important information about the composition, distribution, ecology and diversity of mosquito species such as establishment of mosquitoes in this region. This new and important infor- mation will help us to correctly control and monitor strategies of disease vectors, such as Ae. caspius, Ae. vexans, Cx. pipiens and Cx. theileri. This information helps us to prevent important vectors from increasing and estab- lishment in this area due to change in human activities and weather; thus, the risk of trans- mission of disease by mosquitoes gets mini- mized. The aim of this study was to determine composition, distribution and some ecologi- cal aspects of mosquitoes in East Azerbaijan Province, Iran which is high medically im- portance from the point of view of the arbo- viral vectors. Materials and Methods Study area East Azerbaijan Province is located in northwestern of Iran between 39º 26´–36º 45´ N latitudes and 45º 5´–48º 22´ E longitudes. The province covers an area of approximate- J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 64 http://jad.tums.ac.ir Published Online: April 27, 2019 ly 47,830km², it has a population of around four million people (Fig. 1, Table 1). This province contains 19 counties. Tabriz City is center of the province which 1360m above sea level. The mean annual rainfall is about 300mm. The average relative humidity changes are from 44%. The averages of the maximum and minimum temperatures are 17.7 °C and 6.8 °C, respectively, and the av- erage temperature is 12.3 °C. The province includes arid and semiarid climates (Fig. 1, Table 1). Mosquito collection, site selection and spe- cies identification For mosquito collection, nine sites with different biotopes in three counties of Ahar, Marand, and Tabriz from East Azerbaijan Prov- ince were selected. Collection of mosquito’s larvae was per- formed in different habitats using the stand- ard dipping technique (using 350ml Clark’s dippers) and whirl pack bags (45) (Table 2). Light trap, mosquito net and aspirator was used for adult mosquito collection. Collection of mosquitoes was carried out during June to Oc- tober 2017. Larvae and adult mosquitoes were identified by morphological characters (46). Physical and biological characteristics of larval habitats In these investigation characteristics of mosquitoes larval habitats was studied. The ecological characteristics containing geograph- ical properties of collecting localities (latitude, longitude and altitude), type of habitat (stag- nant, stream, seepage and water container), veg- etation situation (presence or lack of vegeta- tion), kind of vegetation (Null, Poaceae, Typhaceae, Dispacea and Acorus species), wa- ter situation (turbid and clear), exposure to sun- light (full, partial sunlight and covered or shad- ed), depth, substrate type (muddy, sandy, rocky and concrete), distance from animal and human houses and physicochemical attributes such as water temperature and pH were recorded visually or using special equipment (9, 47). Data analysis The species richness (R: Margalef index), unified indices (D: Simpson’s diversity index and H̕: Shannon–Wiener index) and evenness (E: distribution of abundances among the spe- cies) as measures of a diversity, were calcu- lated for East Azerbaijan Province and dif- ferent study counties. The formulae and their rationale in the present study are summarized below: Results Totally, 1401 mosquitoes were collected from nine sites in East Azerbaijan Province during June–October 2017, including 1015 adult and 386 larvae. The properties of geo- graphical larval habitats (Latitude, Longitude and altitude) have been shown in (Fig. 1, Table 1). Four genera along with ten species were collected and identified, including An. hyrca- nus, An. maculipennis s.l., An. superpictus s.l., Ae. caspius, Ae. vexans, Cx. pipiens, Cx. theil- eri, Cx. perexiguus, Cs. longiareolata and Cs. subochrea (Table 2). To our knowledge based on literature, for the first time four species of, Ae. caspius, Ae. vexans, Cx. perexiguus, and Cs. subochrea spe- cies from Ahar and two species of Ae. vexans and Cx. perexiguus were reported from Tabriz County. Considering that no investigations have been already carried out in Marand, all the col- lected species comprising An. hyrcanus, An. maculipennis s.l., Ae. vexans, Cx. pipiens and J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 65 http://jad.tums.ac.ir Published Online: April 27, 2019 Cs. longiareolata (Table 2) are new report for the region. Anopheles maculipennis s.l., Ae. caspius, Ae. vexans, Cx. pipiens, Cx. theileri, Cs. longi- areolata and Cs. subochrea species were col- lected at both adult and larval stages but An. hyrcanus, An. superpictus s.l., and Cx. perex- iguus species were collected only at the lar- val stages (Table 2). In adult stage, Cx. pipiens (37.3%), Cx. theileri (23.8%), Cs. longiareolata (21.8%), Ae. caspius (11.14%), Ae. vexans (5.18%) were the most abundance species respectively, How- ever, An. maculipennis s.l. (0.52%) and Cs. subochrea (0.26%) were the least abundance species (Table 2). At the larval stage Cx. theileri (50.72%), Cs. longiareolata (32.12%) and Cx. pipiens (9.75%) were the most prevalent species re- spectively but in contrast Ae. caspius (2.27%), Cx. perexiguus (1.97%), Cs. subochrea (1.09 %), An. maculipennis s.l., (0.69%), An. hyr- canus (0.5%) and An. superpictus s.l. (0.39%), were the least abundance species respectively (Table 2). Species such as Cx. pipiens, Cx. theileri, Cs. longiareolata and Ae. vexans have a wide distribution in the study areas but some spe- cies such as An. hyrcanus, An. maculipennis s.l., An. Superpictus s.l., and Cs. subochrea have been collected from limited areas (Ta- ble 2). Characteristics of mosquito larval habitats of mosquitoes In Ahar County located in northeastern East Azerbaijan Province, there were three larval sites such as Yavarkandi, Razin and Sattar Khan Dam (Table 3, Fig. 1). Sattar Khan Dam and Razin larval sites had seepage and stag- nant water respectively. Both of the larval sites had turbid water with muddy substrate and shallow depth. In addition, these sites were cov- ered from sunlight with Null, Poaceae, Typhaceae and Dispacea vegetation. Those were more than two kilometers away from human and animal houses. The Sattar Khan Dam larval habitat had 200 meters distance from animal space and its field temperature was 13 °C. In contrast, Razin larval site had 500 meters distance from animal space and its field temperature was 9 °C. In these two larval hab- itats harbored Ae. caspius, Ae. vexans, Cx. theileri, Cx. pipiens, Cx. perexiguus, Cs. subo- chrea and An. superpictus s.l. (Table 2). Ya- varkandi larval site had stagnant and clear wa- ter without vegetation, with muddy substrate and exposed to sunlight. It was more than two km away from human and animal houses. In the Yavarkandi larval site the field tempera- ture was 23 °C. In this larval habitat, two spe- cies of Cs. longiareolata and Cx. pipiens were collected (Table 3). Tabriz larval site is located in the central East Azerbaijan Province and contains three regions of Hojjaj Park, Khaje-dizaj and Chavan (Table 3, Fig. 1). Both regions of Hojjaj Park and Khaje-dizaj had stagnant and turbid wa- ter with muddy substrate. The Hojjaj Park had the following conditions: the parts of larval habitat were covered with Poaceae plants and its water depth was shallow, also it had more than two kilometers distance from animal and human houses. The field temperature was 17 °C in this site (Table 3). This site contains Ae. caspius, Cx. theileri, Cx. pipiens, Cx. perex- iguus and Cs. longiareolata (Table 2). The Khaje-dizaj site was covered with Null and Ziziphu plants. Its depth was less than one meter and its field temperature was 16 °C. It had one-kilometer distance from human houses and about 100 meters from animal sites (Ta- ble 3). In this larval habitat, Cx. theileri and Cx. pipiens were collected (Table 2). Chavan larval habitat was a water container with sub- strate cement and clear water that had no plants and was exposed to sunlight. It had a very short distance with human and animal houses (About 2–10m), which had one meter depth and 27 °C (Table 3). In this larval habitat Cs. longiareolata and Cx. pipiens were collected (Table 2). J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 66 http://jad.tums.ac.ir Published Online: April 27, 2019 In Marand with three larval sites of Ghareh Tappeh, Dolat Abad and Zinab located in north- west East Azerbaijan Province (Fig. 1). Ghareh Tappeh and Dolat Abad habitats had stagnant and clear water. Ghareh Tappeh site had no plants with sandy substrate and shallow depth that was exposed to sunlight. It had one meter depth and 18 °C field temperature (Table 3). Culiseta longiareolata, Cx. pipiens, Ae. vexans, An. maculipennis s.l. and An. hyrcanus were collected in this larval habitat (Table 2). Dolat Abad site was shallow with muddy substrate and 18 °C field temperature. The parts of larval habitat were covered with Null and Dispacea plants and it had more than two kilometers dis- tance from human and animal houses (Table 3). In this larval habitat Cx. theileri and Cx. pipiens were collected (Table 2). Zinab site was stream and clear water with rocky sub- strate and shallow depth. It had no plants and exposed to sunlight with 17 °C temperature. It had about 100 meters distance from human and animal houses (Table 3). In this larval habi- tat, specimens of Cx. pipiens, Cs. longiareo- lata, An. maculipennis s.l. and An. hyrcanus were collected (Table 2, Fig. 1). According to Table 2 and 3, larvae of mos- quitoes occupied all different types of the hab- itats. In addition, our results showed that Cx. pipiens, Cs. longiareolata and Cx. theileri, re- spectively, had the most distribution and ad- aptation to different types of larval habitats. These three species were collected in most larval habitats but in contrast, Cx. perexiguus was found only in the shallow water that was stagnant, turbid, covered with plants and with muddy substrate. In addition, distribution of Cs. subochrea was limited and only was col- lected in the seepage, stagnant and turbid wa- ter habitats with muddy substrate and shallow depth. Anopheles was found only in stream, stagnant and clean water with muddy sub- strate that was exposed to sunlight. Aedes vexans and Ae. caspius were found mostly in larval habitats with turbid water and they were covered with different types of plants. Only two Ae. vexans specimens were found in clean water that was exposed to sunlight with muddy substrate (Table 2, 3). In our study, the temperature and pH ranges were 9 °C to 27 °C and 7–8 respectively. There were differences in the species di- versity, as indicated by the values of Simp- son’s diversity index, Shannon- Wiener index (H’), and evenness and species richness of the mosquito fauna among the study areas of East Azerbaijan Province (Table 4). The species richness and the three indices were found to be maximal in Ahar County (R: 1.5, D: 0.79, H’: 1.74, E: 0.73), whereas the estimated di- versity (D: 0.49, H’: 1.01), and richness (R: 0.82) were the lowest in Marand county. Table 1. Geographical characters of the locations were studied No. Locations Latitude (N) Longitude (E) Altitude (M) 1 Ghareh Tappeh 38• 26' 9.758'' 45• 35' 7.828'' 1296 2 Dolat Abad 38• 45' 53.988'' 45• 49' 19.418'' 1297 3 Zinab 38• 9' 9.654'' 45• 52' 53.218'' 1302 4 Khaje dizaj 38• 12' 21.030'' 46• 16' 39.659'' 1325 5 Hojjaj Park 38• 11' 16.210'' 46• 23' 45.659'' 1336 6 Chavan 37• 59' 9.952'' 46• 23' 32.164'' 1759 7 Sattar Khan Dam 38• 46' 15.552'' 46• 8' 22.763'' 1429 8 Razin 34• 24' 53.932'' 47• 8' 24.629'' 1435 9 Yavarkandi 38• 38' 45.622'' 47• 21' 97.143'' 1706 J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 67 http://jad.tums.ac.ir Published Online: April 27, 2019 Table 2. Details of mosquitoes collected from larval and adult habitats in East Azerbaijan Province during June– October 2017 County Location species Number Total Percent Larvae Adult Ahar Yavarkandi Cs. longiareolata 47 0 47 23 Cx. pipiens 28 4 28 14 Cx. theileri 0 2 2 1 Razin Ae. caspius 7 0 7 3.5 Ae. vexans 3 6 9 4.5 Cx. pipiens 6 10 16 8 Cx. theileri 15 7 22 11 Cx. perexiguus 13 0 13 6.5 An. superpictus s.l. 4 0 4 2 Sattar Khan Dam Cs. subochrea 12 0 12 6 Cx. theileri 34 0 34 17 Cx. pipiens 4 3 7 3.5 Subtotal 173 32 205 100 Tabriz Hojjaj park Ae. caspius 16 43 59 7.76 Cs. longiareolata 13 1 14 1.85 Cx. pipiens 10 17 27 3.55 Cx. theileri 15 12 27 3.55 Cx. perexiguus 7 0 7 .93 Cs. subochrea 0 1 1 .13 Khage dizaj Cx. pipiens 23 73 96 12.63 Cx. theileri 417 39 456 60 Ae. vexans 0 1 1 .13 Chavan Cx. pipiens 11 13 24 3.16 Cs. longiareolata 48 0 48 6.31 Subtotal 560 200 760 100 Marand Ghareh Tappeh Cs. longiareolata 186 83 269 61.6 Cx. pipiens 3 2 5 1.4 Ae. vexans 2 13 15 3.3 An. maculipennis s.l. 2 2 4 .9 An. hyrcanus 1 0 1 .2 Cx. theileri 0 17 17 3.9 Dolat Abad Cx. theileri 34 9 43 9.8 Cx. pipiens 6 14 20 4.6 Zinab Cs. longiareolata 32 0 32 7.3 Cx. pipiens 7 8 15 3.3 Cx. theileri 0 6 6 1.4 An. maculipennis s.l. 5 0 5 1.4 An. hyrcanus 4 0 4 .9 Subtotal 282 154 436 100 Total 1015 386 1401 100 J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 68 http://jad.tums.ac.ir Published Online: April 27, 2019 Table 3. Characteristics of mosquito larval collection sites in Ahar, Tabriz and Marand counties of East Azerbaijan Province during June–October 2017. County Location Type Turbid Exposed Vegetation DHH DAH Depth Sub. T Ahar Yavarkandi Stagnant Clear Exposed Without >2km >2km <1m Muddy 23 Razin Stagnant Turbid Covered Null, Poaceae, Typhaceae, Dispacea >2km 500m Shallow Muddy 9 Sattar Khan Dam Seepage water Turbid Covered Null, Dispacea >2km 200m Shallow Muddy 13 Tabriz Hojjaj park Stagnant Turbid Partial Poaceae 20m >2km Shallow Muddy 17 Khage dizaj Stagnant Turbid Covered Null, Acorus species 1km 100m <1m Muddy 16 Chavan Water container Clear Exposed Without 2m 10m 1m Concrete 27 Marand Ghareh Tappeh Stagnant Clear Exposed Without ND ND 1m Sandy 18 Dolat Abad Stagnant Clear Partial Null, Dispacea >2km >2km Shallow Muddy 19 Zinab Stream Clear Exposed Without 100m 100m Shallow Rocky 17 ND: not determined, Tur: Turbidity, Exp: Sun exposure, Veg: Vegetation type, DHH: Distance from the nearest human houses, DAH: Distance from the nearest animal house, Sub: Substrate type, T: Tempera- ture. Table 4. The species richness (R), Simpson's diversity index (D), Shannon-Weiner diversity index (H̕), and even- ness (E) of the collected mosquito species in East Azerbaijan Province during June–October 2017 Loc. No. S R D H̕ E Marand 436 6 0.823 0.491 1.008 0.563 Tabriz 760 7 0.905 0.547 1.069 0.550 Ahar 205 9 1.503 0.789 1.735 0.727 East Azerbaijan Province 1401 11 1.380 0.694 1.513 0.631 Discussion This research is the first study on distri- bution, diversity and ecology of mosquitoes, with emphasis on Ae. capius and A. vexans as potential arbovirus vectors in East Azerbaijan Province, northwestern of Iran. The East Azer- baijan Province contains diverse geographical areas with different weather conditions. These diverse conditions can provide suitable envi- ronment for the establishment of different spe- cies of mosquitoes and justify the variety of mosquito species in this region. This study showed many mosquito species had ecologi- cal adaptations. In spite of these ecobiologi- cal characteristics, the ecology of mosquitoes present in East Azerbaijan Province is large- ly unknown. In this investigation we tried to study distribution and ecology of mosquitoes in three northeastern, northwest and central regions of East Azerbaijan Province, where various mosquito vectors of malaria and ar- boviruses are present (48-49). Some studies had been conducted on fauna and checklist of mosquitoes in parts of this region (48), alt- hough to our knowledge no studies have been done on the ecology of mosquitoes in these regions. J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 69 http://jad.tums.ac.ir Published Online: April 27, 2019 In current study, 4 genera and 10 species were collected and identified. For the first time, Ae. caspius, Ae. vexans, Cx. perexiguus, Cs. subochrea species were reported from Ahar county. Also for the first time, Ae. vexans and Cx. perexiguus species were reported from Tabriz county. As well as for the first time, An. hyrcanus, An. maculipennis s.l., Ae. vexans, Cx. pipiens, Cx. theileri and Cs. longiareola- ta species were reported from Marand coun- ty. Some previous studies had found only An. hyrcanus in northwestern Iran (49-50) and oth- er study identified An. claviger, An. hyrcanus, An. maculipennis s.l., An. pseudopictus, An. sacharovi and An. Superpictus s.l. in East Azer- baijan Province. In the present study, we re- ported An. hyrcanus, An. maculipennis s.l. and An. Superpictus s.l.. In previous studies, Cx. pipiens and Cx. theileri were reported in these regions (48), but Cx. perexiguus was not found in these regions. In this study, three species of Culex genus were reported such as Cx. pipiens, Cx. theileri and Cx. perexiguus that Cx. pipiens and Cs. longiareolata species were the most abundant. Aedes caspius and Ae. vexans were found only from the Kaleybar region (49), In our study, for the first time, Ae. caspius and Ae. vexans were found in Ahar, Marand and Tabriz, where previous study did not report these species in these areas (48). One study reported Cs. annulata and Cs. subochrea in West Azerbaijan Province and Cs. subochrea in East Azerbaijan Province (51). Culiseta longiareolata species was reported as the most abundant in Kermanshah, Kurdistan and Sis- tan and Baluchistan provinces (52). In this study, two members of Culiseta genus were found, including Cs. longiareolata and Cs. subochrea where Cs. longiareolata species was the most abundant species. Comparing the results of our study with a recent study in West Azerbaijan Province (47) showed that six species An. maculipennis s.l., An. superpictus s.l., Cx. pipiens, Cx. theil- eri, Cs. longiareolata and Ae. caspius were common between West and East Azerbaijan provinces. The results of our study compared with the results of research conducted in Zan- jan Province (53), showed that five species (An. maculipennis s.l., An. superpictus s.l., Cx. pipiens, Cx. theileri, and Cs. longiareolata) were common between these two provinces. The comparison the results of our study with a recent study conducted in Kurdistan Prov- ince (54) showed that six species (An. macu- lipennis s.l., An. superpictus s.l., Cx. theileri, Cx. pipiens, Cs. longiareolata and Ae. caspius) were common between in provinces. In Tur- key (55), and in the provinces of Ardebil, Kur- distan and West Azarbaijan, Cx. theileri and Cx. pipiens were dominant and most abundant species (56) and our study results confirm this information. Our results showed that Cx. theil- eri (50.72%), Cs. longiareolata (32.12%) and Cx. pipiens (9.75%) were dominant and the most abundant species in this province. The climate changes and biotic and abiotic environment factors including plants, temper- ature and rainfall ranges, significantly affect the type and frequency of larval mosquito habitats. These factors, in turn, affect the number of mos- quito species, larval stages, longevity, behav- ior and adult development. As a result, the transmission of diseases through mosquitoes is directly affected by environmental factors (57- 58). The presence of plants as a source of sugar for mosquitoes is very important, that influence both larval and adult stage development (59- 70). Plants provide energy for mosquitoes, As a result, survival rate increased and longevity of the mosquito longer than the extrinsic incu- bation period of parasite, therefore the inci- dence of disease increases (61-62). In our study, five types of plants such as Null, Poaceae, Typhaceae, Carex dipsacea and Acorus spe- cies were found in relation to mosquito larval habitats. To our knowledge, there is no study on plant species associated with mosquito spe- cies in Iran. In our study, various species of mosquitoes such as Anopheles, Culex, Aedes and Culisita were found in relation with var- J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 70 http://jad.tums.ac.ir Published Online: April 27, 2019 ious plants, therefore this study confirms the results of previous research. Aedes vexans and Ae. caspius have not been reported earlier in these areas. These results are likely to show the effects of climate changes and human activities on the distribution of these species in these re- gions. Physicochemical properties of larval habi- tats regulate the abundance of mosquito species, for example, Anopheles species were found more in natural larval habitats and Culex spe- cies in artificial larval habitats (63). In our study, Anopheles species were found in natu- ral habitats, while Culex species were collected from different types of larval habitats, indicat- ing these mosquitoes can live in a wide range of water habitats. Larval habitats in this study were stagnant, stream and seepage, water con- tainer, turbid and clear water, sun exposed or covered from sunlight –rocky or muddy sub- strate and shallow depth. Studies showed that some of Culex larval species were found alone or along with other mosquitoes, such as Anophele and Aedes (64-67) which has been observed in our study. There was a significant relationship be- tween the distance of larval habitats of Anoph- eles mosquitoes, human and animal sites, be- sides, Anopheles mosquitoes are found more often near human and animal houses (68). Our study showed that there was a significant cor- relation between the distances of larval habi- tats of Anopheles, Ae. caspius, Ae. vexans and Cx. theileri mosquitoes with human and an- imal sites, because the larval habitats of these mosquitoes were found more near to human and animal houses. However, this correlation was not observed between the larval habitats of Cx. pipiens and Cs. longiareolata with hu- man and animal houses, because the habitats of these mosquitoes were found at various intervals from human and animal houses. Conclusion Arbovirus vectors such as Ae. caspius and Ae. vexans along with Cx. pipiens and Cx. theil- eri are well adapted to a broad range of habi- tats and climatic conditions. Determining of distribution and full description of ecology of arboviral vectors under local eco-demographic conditions in the East Azerbaijan Province have provided important ecological information on establishment of important mosquito borne diseases. This new information will help us to correctly control and monitor strategies of disease vectors, and in this way prevent im- portant vectors such as ae. Ae. Caspius, Ae. vexans, Cx. pipiens and Cx. theileri to be in- creased and established in this area due to changing human activities and weather chang- es. These strategies help minimizing the risk of transmission of disease by mosquitoes. Acknowledgements This paper is a part of the results of the first author’s dissertation for fulfillment of a PhD degree in Medical Entomology and Vec- tor Control from Department of Medical En- tomology and Vector Control, School of Pub- lic Health, Tehran University of Medical Sci- ences, Tehran, Iran. This study was financially supported by the Deputy for Research, Tehran University of Medical Sciences Project no. 35904. The authors declare that there is no con- flict of interests. References 1. Dong X, Dong L, Wu C (2008) A new species of the genus Tripteroides (Dip- tera: Culicidae) from China. Acta Zoo Taxon Sin. 33: 187–190. 2. Wilkerson RC, Linton YM, Fonseca DM, Schultz TR, Price DC, Strickman DA (2015) Making mosquito taxonomy use- ful: A stable classification of tribe Ae- http://mosquito-taxonomic-inventory.info/biblio?f%5Bauthor%5D=135 http://mosquito-taxonomic-inventory.info/biblio?f%5Bauthor%5D=136 http://mosquito-taxonomic-inventory.info/biblio?f%5Bauthor%5D=137 J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 71 http://jad.tums.ac.ir Published Online: April 27, 2019 dini that balances utility with current knowledge of evolutionary relationships. PLoS One. 10(7): e0133602. 3. Dong X, Zhou H, Gong ZD, Lu BL (2004) Investigation of mosquito species in Yun- nan Province with some new species. Chinese J Vector Biol. 15: 186–191. 4. Reinert JF, Harbach R, Kitching IJ (2009) Phylogeny and classification of tribe Aedini (Diptera: Culicidae). Zool J Linn Soc. 157(4): 700–794. 5. Lemine AM, Lemrabott MA, Ebou MH, Lekweiry KM, Salem MS, Brahim KO, Moukah MO, Bouraya IN, Brengues C, Trape JF, Basco L (2017) Mosquitoes (Diptera: Culicidae) in Mauritania: a re- view of their biodiversity, distribution and medical importance. Parasit Vectors. 10(1): 35. 6. WHO (2017) World malaria report 2017. WHO Press, Geneva, Switzerland. 7. Davis LE, Beckham JD, Tyler KL (2008) North American encephalitic arbovirus- es. Neurol Clin. 26(3): 727–757. 8. Fontenille D, Traoré-Lamizana M, Zeller H, Mondo M (1995) Rift Valley fever in western Africa: isolations from Aedes mosquitoes during an interepizootic pe- riod. Am J Trop Med Hyg. 52: 403–404. 9. Khater EI, Sowilem MM, Sallam MF, Alahmed AM (2013) Ecology and hab- itat characterization of mosquitoes in Saudi Arabia. Trop Biomed. 30: 409– 427. 10. Hanafi-Bojd AA, Vatandoost H, Oshaghi MA, Charrahy Z, Haghdoost AA, Sed- aghat MM, Abedi F, Soltani M, Raeisi A (2012) Larval habitats and biodiver- sity of anopheline mosquitoes (Diptera: Culicidae) in a malarious area of south- ern Iran. J Arthropod Borne Dis. 49(2): 91–100. 11. Hanafi-Bojd AA, Vatandoost H, Oshaghi MA, Charrahy Z, Haghdoost A, Zamani G, Abedi F, Sedaghat MM, Soltani M, Shahi M, Raeisi A (2012) Spatial anal- ysis and mapping of malaria risk in an endemic area, south of Iran: a GIS based decision making for planning of control. Acta Trop. 122(1): 132–137. 12. Blaustein L, Chase JM (2007) Interactions between mosquito larvae and species that share the same trophic level. Annu Rev Entomol. 52: 489–507. 13. Juliano SA (2009) Species interactions among larval mosquitoes: context de- pendence across habitat gradients. An- nu Rev Entomol. 54: 37–56. 14. Khodzhaeva LF, Issi IV (1989) A new ge- nus of microsporidians, Cristulospora gen. n. (Amblyosporidae) with 3 new species from blood-sucking mosquitoes from Uzbekistan. Parazitologiya. 23(2): 140–145 (In Russian). 15. Detinova TS, Smelova VA, Voprosuo K (1973) Medical importance of mosqui- toes (Culicidae, Diptera) from the fau- na of the Soviet Union. Med Parazitol. 42(4): 455–4471 (In Russian). 16. Nahla M, Belal A, Doaa FY, Ismail M (2013) Rapid assay to differentiate the two forms of Egyptian Aedes caspius (Diptera: Culicidae) using acetylcholin- esterase gene. J Basic Appl Zool. 66(1): 12–17. 17. Cranston PS, Ramsdale CD, Snow KR, White GB (1987) Adults, larvae and pu- pae of British mosquitoes (Culicidae). Freshwater Biological Association, Am- bleside. 18. Bagheri M, Terenius O, Oshaghi MA, Motazakker M, Asgari S, Dabiri F, Vatandoost H, Mohammadi Bavani M, Chavshin AR (2015) West Nile Virus in mosquitoes of Iranian wetlands. Vec- tor Borne Zoonotic Dis. 15(12): 750–754. 19. Knight KL, Stone A (1977) A catalog of the mosquitoes of the world (Diptera: Culicidae). Vol. 6. Entomological So- ciety of America, Maryland. https://www.ncbi.nlm.nih.gov/pubmed/4149080 J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 72 http://jad.tums.ac.ir Published Online: April 27, 2019 20. Smith KGV (1973) Insects and other Ar- thropods of medical importance. British Museum (Natural History), London. 21. Mith CEG (1987) Factors influencing the transmission of western equine enceph- alomyelitis virus between its vertebrate maintenance hosts and from them to humans. Am J Trop Med Hyg. 37: 33–39. 22. Horsfall WR (1955) Mosquitoes, their bi- onomics and relation to disease. Hafner Publishing, New York. 23. Ludlam KW, Jachowski LA, Otto GF (1970) Potential vectors of Dirofilaria immitis. J Am Vet Med Assoc. 157: 1354–1359. 24. Vatandoost H, Oshaghi MA, Abaie MR, Shahi M, Yaaghoobi F, Baghaii M (2006) Bionomics of Anopheles stephensi Lis- ton in the malarious area of Hormozgan Province, southern Iran, 2002. Acta Trop. 97(2): 196–203. 25. Koosha M, Oshaghi MA, Sedaghat MM, Vatandoost H, Azari-Hamidian S, Abai MR, Hanafi-Bojd AA, Mohtarami F (2017) Sequence analysis of mtDNA COI barcode region revealed three hap- lotypes within Culex pipiens assemblage. Exp Parasitol. 181: 102–110. 26. Salim Abadi Y, Vatandoost H, Sanei Dehkordi AR, Paksa A (2010) Evalua- tion of biological control agents for mos- quitoes control in artificial breeding plac- es. Asian Pac J Trop Dis. 3(4): 276–277. 27. Soleimani-Ahmadi M, Abtahi SM, Madani A, Paksa A, Salim Abadi Y, Gorouhi MA, Sanei-Dehkordi, A (2017) Phyto- chemical profile and mosquito larvicid- al activity of the essential oil from aer- ial parts of Satureja bachtiarica Bunge against malaria and lymphatic filariasis vectors. J Essent Oil Bear. 20(2): 328– 336. 28. Nikookar SH, Moosa-Kazemi SH, Oshaghi MA, Yaghoobi-Ershadi MR, Vatandoost H (2010) Species composition and di- versity of mosquitoes in Neka County, Mazandaran Province, northern Iran. Iran J Arthropod Borne Dis. 4(2): 26–34. 29. Karami M, Moosa-Kazemi SH, Oshaghi MA, Vatandoost H, Sedaghat MM, Ra- jabnia R, Hosseini M, Maleki-Ravasan N, Yahyapour Y, Ferdosi-Shahandashti E (2016) Wolbachia Endobacteria in natural populations of Culex pipiens of Iran and its phylogenetic congruence. J Arthropod Borne Dis. 10(3): 347–363. 30. Nikookar SH, Moosa-Kazemi SH, Yaghoobi-Ershadi MR, Vatandoost H, Oshaghi MA, Ataei A, Anjamrooz M (2015) Fauna and Larval Habitat Charac- teristics of mosquitoes in Neka County, Northern Iran. J Arthropod Borne Dis. 9 (2): 253–266. 31. Nikookar SH, Moosa-Kazemi SH, Oshaghi MA, Vatandoost H, Yaghoobi-Ershadi MR, Enayati AA, Motevali-Haghi F, Zia- pour SP, Fazeli-Dinan M (2015) Biodi- versity of culicid mosquitoes in rural Ne- ka township of Mazandaran Province, northern Iran. J Vector Borne Dis. 52(1): 63–72. 32. Soleimani-Ahmadi M, Gorouhi MA, Aza- ni S, Salim Abadi Y, Paksa A, Rashid G, Sanei-Dehkordi, A (2017) Larvicidial Effects of essential oil and methanol ex- tract of Achillea wilhelmsii C. Koch (Asteraceae) against Anopheles stephen- si Liston (Diptera: Culicidae), a malaria vector. J Kerman Univ Med Sci. 24(1): 58–67. 33. Chavshin AR, Oshaghi MA, Vatandoost H, Hanafi-Bojd AA, Raeisi A, Nikpoor F (2014) Molecular characterization, bi- ological forms and sporozoite rate of Anopheles stephensi in southern Iran. Asian Pac J Trop Biomed. 4(1): 47–51. 34. Mehravaran A, Vatandoost H, Oshaghi MA, Abai MR, Edalat H, Javadian E, Mashayekhi M, Piazak N, Hanafi-Bojd AA (2012) Ecology of Anopheles ste- phensi in a malarious area, southeast of Iran. Acta Med Iran. 50(1): 61–65. J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 73 http://jad.tums.ac.ir Published Online: April 27, 2019 35. Vatandoost H, Emami SN, Oshaghi MA, Abai MR, Raeisi A, Piazzak N, Mahmoodi M, Akbarzadeh K, Sartipi M (2011) Ecology of malaria vector Anoph- eles culicifacies in a malarious area of Sistan va Baluchestan Province, south- east Islamic Republic of Iran. East Med- iterr Health J. 17(5): 439–445. 36. Oshaghi MA, Vatandoost H, Gorouhi A, Abai MR, Madjidpour A, Arshi S, Sadeghi H, Nazari M, Mehravaran A (2011) Anopheline species composition in borderline of Iran-Azerbaijan. Acta Trop. 119(1): 44–49. 37. Oshaghi MA, Yaghobi-Ershadi MR, Shemshad K, Pedram M, Amani H (2008) The Anopheles superpictus com- plex: introduction of a new malaria vec- tor complex in Iran. Bull Soc Pathol Exot. 101(5): 429–434. 38. Oshaghi MA, Shemshad Kh, Yaghobi- Ershadi MR, Pedram M, Vatandoost H, Abaie MR, Akbarzadeh K, Mohtarami F (2007) Genetic structure of the malaria vector Anopheles superpictus in Iran us- ing mitochondrial cytochrome oxidase (COI and COII) and morphologic mark- ers: a new species complex? Acta Trop. 101(3): 241–248. 39. Oshaghi MA, Yaaghoobi F, Abaie MR (2006) Pattern of mitochondrial DNA variation between and within Anopheles stephensi (Diptera: Culicidae) biological forms suggests extensive gene flow. Ac- ta Trop. 99(2–3): 226–233. 40. Naddaf SR, Oshaghi MA, Vatandoost H, Assmar M (2003) Molecular character- ization of Anopheles fluviatilis species complex in the Islamic Republic of Iran. East Mediterr Health J. 9(3): 257–265. 41. Oshaghi MA, Sedaghat MM, Vatandoost H (2003) Molecular characterization of the Anopheles maculipennis complex in the Islamic Republic of Iran. East Med- iterr Health J. 9(4): 659–666. 42. Mehravaran A, Oshaghi M, Vatandoost H, Abai M, Ebrahimzadeh A, Roodi AM, Grouhi A (2011) First report on Anoph- eles fluviatilis U in southeastern Iran. Acta Trop. 117(2): 76–81. 43. Doosti S, Azari-Hamidian S, Vatandoost H, Oshaghi MA, Hosseini M (2006) Taxonomic differentiation of Anophe- les sacharovi and An. maculipennis s.l. (Diptera: Culicidae) larvae by seta 2 (antepalmate hair). Acta Med Iran. 44 (1): 21–27. 44. Karimian F, Oshaghi MA, Sedaghat MM, Waterhouse RM, Vatandoost H, Hanafi- Bojd AA, Ravasan NM, Chavshin AR (2014) Phylogenetic analysis of the ori- ental-Palearctic-Afrotropical members of Anopheles (Culicidae: Diptera) based on nuclear rDNA and mitochondrial DNA characteristics. Jpn J Infect Dis. 67(5): 361–367. 45. Silver JB (2008) Mosquito ecology: field sampling methods. 3rd ed. Dordrecht, Netherlands. 46. Azari-Hamidian S, Harbach RE (2009) Keys to the adult females and fourth- instar larvae of the mosquitoes of Iran (Diptera: Culicidae). Zootaxa. 2078: 1– 33. 47. Khoshdel-Nezamiha F, Vatandoost H, Azari-Hamidian S, Mohammadi Bavani M (2014) Fauna larval habitats of mos- quitoes (Diptera: Culicidae) of West Azerbaijan Province, Northwestern Iran. J Arthropod-Borne Dis. 8(2): 163–173. 48. Abai MR, Azari-Hamidian S, Hakimi M, Mashhadi-Esmai K, Vatandoost H (2007) Fauna and checklist of mosquitoes (Dip- tera: Culicidae) of East Azerbaijan Prov- ince,Northwestern Iran. Iran J Arthro- pod Borne Dis. 1(2): 27–33. 49. Vatandoost H, Abdoljabari Boonab R, Abai MR, Oshaghi MA (2005) Entomological survey in Kalibar, a resurgent malaria focus in East-Azerbaijan, Iran Pak J Biol Sci. 8: 1466–1471. J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 74 http://jad.tums.ac.ir Published Online: April 27, 2019 50. Yaghoobi-Ershadi MR, Namazi J, Piazak N (2001) Bionomics of Anopheles sa- charovi in Ardabil Province, northwest- ern Iran during a larval control program. Acta Trop. 78: 207–215. 51. Kalandadze LP, Kaviladze OP (1947) On the blood-sucking mosquitoes of the western part of the Iran Azerbaijan. Med Parasitol Parasitic Dis. 16: 57–65 (In Rus- sian). 52. Moosa-Kazemi H, Zahirnia AH, Sharifi F, Davari B (2015) The Fauna and Ecology of Mosquitoes (Diptera: Culicidae) in Western Iran. J Arthropod Borne Dis. 9 (1): 49–59. 53. Ghavami MB, Ladonni H (2005) The fau- na and frequency of different mosquito species (Diptera: Culicidae) in Zanjan Province. J Zanjan Univ Med Sci Health Serv. 13(53): 46–54. 54. Banafshi O, Abai MR, Ladonni H, Bakhsh H, Karami H, Azari-Hamidian S (2013) The fauna and ecology of mosquito lar- vae (Diptera: Culicidae) in western Iran. Turk J Zool. 37: 298–307. 55. Aldemir A, Bedir H, Demirci B, Alten B (2010) Biting activity of mosquito spe- cies (Diptera: Culicidae) in the Turkey- Armenia border area, Ararat Valley, Tur- key. J Med Entomol. 47(1): 22–27. 56. Azari-Hamidian S, Yaghoobi-Ershadi MR, Javadian E, Abai MR, Mobedi I, Linton YM, Harbach RE (2009) Distribution and ecology of mosquitoes in a focus of diro- filariasis in northwestern Iran, with the first finding of filarial larvae in natural- ly infected local mosquitoes. Med Vet Entomol. 23(2): 111–121. 57. Murdock CC, Paaijmans KP, Cox-Foster D, Read F (2012) Rethinking vector im- munology: the role of environmental tem- perature in shaping resistance. Nat Rev Microbiol. 10: 869–876. 58. Oshaghi MA, Ravasan NM, Javadian E, Rassi Y, Sadraei J, Enayati AA, Vatan- doost H, Zare Z, Emami SN (2009) Ap- plication of predictive degree day mod- el for field development of sandfly vec- tors of visceral leishmaniasis in north- west of Iran. J Vector Borne Dis. 46(4): 247–255. 59. Ye-Ebiyo Y, Pollack RJ, Kiszewski A, Spielman A (2003) Enhancement of de- velopment of larval Anopheles arabiensis by proximity to flowering maize (Zea mays) in turbid water and when crowd- ed. Am J Trop Med Hyg. 68(6): 748–752. 60. Müller GC, Beier JC, Traore SF (2010) Field experiments of Anopheles gambiae attraction to local fruits/seedpods and flowering plants in Mali to optimize strat- egies for malaria vector control in Afri- ca using attractive toxic sugar bait meth- ods. Malar J. 20(9): 262. 61. Kebede A, McCann JC, Kiszewski AE, Ye- Ebiyo Y (2005) New evidence of the ef- fects of agro-ecologic change on malaria transmission. Am J Trop Med Hyg. 73 (4): 676–680. 62. Gu W, Müller G, Schlein Y, Novak RJ, Beier JC (2011) Natural plant sugar sources of Anopheles mosquitoes strong- ly impact malaria transmission potential. PLoS One. 6(1): e15996. 63. Alahmed AM, Kheir SM, Kuriji MA, Sallam MF (2011a) Breeding habitats characterization of Anopheles mosquito (Diptera: Culicidae) in Najran Prov- ince, Saudi Arabia. J Egypt Soc Parasi- tol. 41(2): 275–288. 64. Abdullah MA, Merdan AI (1995) Distri- bution and ecology of the mosquito fau- na in the southwestern Saudi Arabia. J Egypt Soc Parasitol. 25(3): 815–837. 65. Alahmed A, Kheir S (2005) Seasonal ac- tivity of some haematophagous insects in the Riyadh region, Saudi Arabia. JSSAS. 4(2): 95–105. 66. Alahmed A, Al Kheriji M, Kheir S (2007) Distribution of habitats of mosquito lar- vae (Diptera: Culicidae) in Riyadh Re- J Arthropod-Borne Dis, March 2019, 13(1): 62–75 A Paksa et al.: Biodiversity of … 75 http://jad.tums.ac.ir Published Online: April 27, 2019 gion, Saudi Arabia. J King Saud Univ Eng Sci. 19: 35–55. 67. Alahmed AM, Al Kuriji MA, Kheir SM, Alahmedi SA, Al Hatabbi MJ, Gashmari MA (2009) Mosquito fauna (Diptera: Cu- licidae) and seasonal activity in Makka Al Mukarramah Region, Saudi Arabia. J Egypt Soc Parasitol. 39(3): 991–1013. 68. Minakawa N, Seda P, Yan G (2002) In- fluence of host and larval habitat distri- bution on the abundance of African ma- laria vectors in western Kenya. Am J Trop Med Hyg. 67(1): 32–38.