J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 350 http://jad.tums.ac.ir Published Online: Dec 31, 2022 Original Article Biodiversity and Spatial Distribution of Mosquitoes (Diptera: Culicidae) in Kurdistan Province, Western Iran Farid Sharifi1, Omid Banafshi1, Ali Rasouli1, Shadi Ghoreishi2, Samrand Saeedi1, Mahmood Khalesi1, Afshar Rezai1, Eslam Moradi Asl3, Bushra Zareie4, Nasrollah Veisi Khodlan1, *Arshad Veysi1 1Zoonoses Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran 2Cellular and Molecular Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran 3Department of Public Health, School of Health, Ardabil University of Medical Sciences, Ardabil, Iran 4Department of Epidemiology, School of Public Health, Hamadan University of Medical Sciences, Hamadan, Iran *Corresponding author: Dr Arshad Veysi, Email: arshadveysi@gmail.com (Received 30 Jan 2022; accepted 17 Dec 2022) Abstract Background: Mosquitoes (Diptera: Culicidae) have always been considered as the vector/s of viral and parasitic dis- eases. This study aimed to conduct a comprehensive survey on the species composition, spatial distribution, and biodi- versity indices of mosquitoes in Kurdistan Province, western Iran. Methods: This study was carried out in 10 counties of Kurdistan Province. The immature stages of mosquitoes were collected monthly from June to September. ArcGIS software was used to spatial analysis and create maps. Alpha diver- sity indices were calculated using the related formula. Results: Totally, 5831 larvae belonging to the family Culicidae were collected. Twelve species were identified includ- ing: Anopheles claviger, An. maculipennis s.l, An. superpictus s.l, Culiseta. longiareolata, Cs. subochrea, Culex horten- sis, Cx. mimeticus, Cx. perexiguus, Cx. pipiens, Cx. theileri, Cx. modestus and Cx. territans. Based on this analysis, the high-risk areas of the province are determined as Anopheles in the west, Culex in the north, and the Culiseta in the south of the province. Analyzing the Alpha biodiversity indices showed Baneh and Sarabad had the maximum and Bijar had the minimum mosquito biodiversity. Conclusion: The western counties of the province are regarded as the hotspots for anopheline mosquitos. Moreover, reporting of malaria cases in the past, bordering with Iraq and the high traffic of travelers have made these areas as po- tential foci for malaria transmission. So that, routine entomological inspections are proposed to detect any suspicious vector or case entrance. Keywords: Mosquitoes; GIS; Diversity; Diptera; Larvae Introduction Mosquitoes (Diptera: Culicidae) have al- ways been the focus of entomological studies due to their important role in transmitting a wide range of viral and parasitic diseases to humans or animals. More than half of the world's popula- tion lives in areas at risk of mosquito-borne dis- eases such as malaria, dengue fever, chikungu- nya, West Nile fever, Japanese encephalitis, and filariasis (1). Malaria is a parasitic infection transmitted by anopheline mosquitoes. It is es- timated that 219 million new cases are report- ed worldwide, and lead to the death of more than 400,000 people annually. Dengue is the most common viral disease transmitted by Ae- Copyright © 2022 The Authors. Published by Tehran University of Medical Sciences. This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International license (https://creativecommons.org/licenses/by- nc/4.0/). Non-commercial uses of the work are permitted, provided the original work is properly cited. http://jad.tums.ac.ir/ https://creativecommons.org/licenses/by-nc/4.0/ https://creativecommons.org/licenses/by-nc/4.0/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 351 http://jad.tums.ac.ir Published Online: Dec 31, 2022 des mosquitoes. More than 3.9 billion people in more than 129 countries are at risk of get- ting dengue fever, with an estimated 96 million symptomatic cases and an estimated 40,000 deaths each year (2). Malaria is also the most important mosquito-borne disease in Iran, which occurs mostly in the southern provinces of the country (3, 4). Seven species of Anopheles are among the most important vectors in Iran, in- cluding: Anopheles stephensi Liston, 1901, An. culicifacies sensu lato (s.l.) Giles, 1901 (5), An. maculipennis s.l. Meigen, 1818 (6), An. fluvi- atilis s.l. James, 1902, An. sacharovi Favre, 1903, An. dthali Patton, 1905, and An. super- pictus s.l. Grassi, 1899 (7, 8). The presence of West Nile and Sindbis viruses, which are trans- mitted by mosquitoes, has also been reported in the country (9–13). Recently, mosquito-borne Dirofilaria (D. immitis and D. repens) has been reported in Iran (14). In recent years, no local transmission of malaria has been reported in Kurdistan Prov- ince, and only in 2020, one case was reported in the city of Marivan, which was Iraqi (un- published data from Department of Disease Control, Deputy of Health, Kurdistan Univer- sity of Medical Sciences). In 2013 serologi- cally positive cases of dengue fever was re- ported from Kurdistan Province, who had not travelled abroad (15); although, there is no re- port of occurring the vector Ae. albopictus or Ae. eagypti in this province. Till now no human case of Rift Valley fever virus has been re- ported in the province, though one study showed infection in sheep (16). West Nile virus is a widespread mosquito-borne arbovirus in Iran as it is reported in horses from at least 26 of the 31 Iranian provinces including Kurdistan (9). Different blood-feeding arthropods including mosquitos are considered as suspected vectors of tularemia which caused by the bacterium Francisella tularensis. Natural infection of tu- laremia in the wild reservoir, rodents (17), and in human have been reported in Kurdistan Prov- ince (18). The latest updated checklist of Ira- nian mosquitoes includes 70 species that rep- resent eight or 12 genera, depending on the gen- eral classification of aedines (19, 20). Zaim (21) mentioned two genera and eight species of the subfamily Culicinae in Kurdistan Province. In previous studies conducted in the province, 18 species of the family belonging to five genera have been reported, which include An. macu- lipennis s.l., An. superpictus s.l., An. claviger, An. sacharovi, An. algeriensis, An. marteri, An. sergentii, Coquillettidia richiardii, Culex hor- tensis, Cx. mimeticus, Cx. pipiens, Cx. theileri, Cx. perexiguus, Cx. territans, Culiseta longi- areolata, Cs. subochrea, Ae. vexans and Ae. caspius (22–24). In recent years, due to the re- porting of dengue vectors in the south of the country (25, 26), and on the other hand, high reporting cases of dengue fever in eastern neigh- bors such as Pakistan, the importance of rou- tine entomological studies has doubled in oth- er border provinces to monitor the entrance of dengue vector/s. Sharing border with Iraq, made Kurdistan Province as one of the most important import destinations for goods from southeast Asian countries. One way of entering the den- gue fever vector/s into the countries is through goods such as bamboo plant and car tires, therefore, conducting entomological studies in this critical point seem necessary. The studies that have been done in the province so far are only faunistic and scattered. The purpose of this study was to conduct a comprehensive study and update the species composition of mosqui- toes in all counties of the province; moreover, to analyze the spatial distribution and to de- termine biodiversity indices of the mosquitoes throughout the province. Materials and Methods Study area This study was conducted in 10 counties of Kurdistan Province in summer 2019. It is in the west of Iran between 34°44′N and 36°28′N and 45°33′E and 48°15′E. The area is 29.137 square kilometers, equivalent to 1/7 of the total area of Iran. This province, which is http://jad.tums.ac.ir/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 352 http://jad.tums.ac.ir Published Online: Dec 31, 2022 located in the scattered slopes and plains of the Middle Zagros Mountains, is limited from the north to the provinces of West Azerbaijan and Zanjan, from the east to Hamedan and Zan- jan, from the south to Kermanshah, and from the west to Iraq. Kurdistan Province has 10 counties include: Sanandaj, Saqez, Marivan, Baneh, Qorveh, Kamyaran, Bijar, Divandarreh, Dehgolan and Sarvabad (Fig. 1). According to the general population and housing census of 2016, Kurdistan Province has a population of 1,603, 011 (Statistical Center of Iran). The high- est and lowest average air temperatures in 2019 are belong to Sanandaj with 14.8 °C and Zar- rineh with 8.8 °C, which they have different height from sea level. Marivan with 913.1mm and Qorveh with 352.7mm had the highest and lowest rainfall, respectively (Statistical Center of Iran). Collection, mounting and identification of mosquito larvae The immature stages of mosquitoes were collected monthly, for at least four months (once a month) from June to September. Larval col- lection was conducted in an urban point and four rural points at a distance of 15–20km in all counties. In fixed urban and rural points, three fixed and three variable locations were sampled. The distribution of sampling points in- cluded urban, rural, cultivated areas, and riv- ersides. The mean water temperature and am- bient humidity were recorded during the sam- pling period. The larvae of the mosquitoes were collected by standard dipping technique. Col- lected larvae were stored in lactophenol me- dium, and after clarification of morphological characters, they were mounted in de Faure’s medium. After that, the larvae were identified using valid morphological Iranian identifica- tion keys (27). Additionally, the geographical coordinates of the sampling locations were recorded using a GPS device. Collecting larvae using Ovitraps Ovitrap is one of the most common tech- niques for collecting eggs and larvae of Aedes mosquitoes. In this study buckets, which were designed in a standard way, were used. Dark buckets, preferably black due to its attractive- ness for Aedes mosquitoes, in a volume of two liters were used. After pouring two liters of wa- ter inside the buckets, 3 wooden pedals with dimensions of 15×5cm and a thickness of one to two mm were immersed in the water, and then they were attached to the wall of the bucket with clamps (28). All the information in the labels of the traps and the exact addresses of their installation places were recorded in a separate booklet. The traps were checked twice a week according to the weather conditions of the study areas. Due to evaporation of water inside the traps, water was added to them if necessary. Dur- ing twice-a week check-up, the pedals were care- fully examined for the presence of eggs by a handy lens. If a suspicious case of eggs was ob- served, the pedals were carefully removed and transported to the laboratory, and it was re- placed by a new pedal. GIS analysis ArcGIS 10.4.1 software (http://www.esri. com/arcgis) was used to spatial analysis and cre- ating maps. After inserting geographic coordi- nates into the Excel software, they were entered into the ArcMap in ArcGIS 10.4.1 software. Inverse Distance Weighted (IDW) interpolation analysis was employed to prepare raster maps. Data process and analysis To determine alpha diversity indices, data were inserted in Microsoft office Excel 2016 software using related indices, and coefficients showing below. Then, the average and stand- ard deviation of all indices were calculated (29,30), and their P-value was determined by performing the analysis of variance (ANO- VA) test in IBM SPSS Statistics 26 software. Shannon–Weiner index: H': species diversity index, S: number of spe- cies, pi: proportion of individuals of each spe- cies belonging to its species of the total num- ber of individuals. http://jad.tums.ac.ir/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 353 http://jad.tums.ac.ir Published Online: Dec 31, 2022 Evenness index: = H': species diversity index, S: number of spe- cies Simpson’s Index: n: the total number of organisms of a particu- lar species, N: the total number of organisms of all species Menhinick’s index: S: number of species, N: the total number of organisms of all species Margalef’s index: S: number of species, N: the total number of organisms of all species Hill: e: Napier's Constant, H’: species diversity in- dex Hil2: pi: proportion of individuals of each species belonging to its species of the total number of individuals. Results Species composition During this study, a total of 5831 larvae belonging to the family Culicidae were col- lected from 56 localities of 10 counties of the province (Table 1–2). Of these, the highest num- ber were from Marivan County and the lowest number were from Kamyaran County (Table 2). The collected larvae belonged to three gen- era: Anopheles 889 (15.24%), Culiseta 1826 (31.31%) and Culex 3116 (53.43). Collecting methods through ovitraps detected only a few larvae, which is belonging to the genus Anoph- eles, and no Aedes species were identified. 12 species of Culicidae were identified includ- ing: An. claviger, An. maculipennis s.l., An. su- perpictus s.l., Cs. longiareolata, Cs. subochrea, Cx. hortensis, Cx. mimeticus, Cx. perexiguus, Cx. pipiens, Cx. theileri, Cx. modestus and Cx. territans (Table 2). Spatial analysis Species belonging to Anopheles were iden- tified in more than 50% of the counties with high abundance. The Baneh County, the western re- gion bordering Iraq, has the highest number of species of this genus, about 2–3 times higher than other counties. Species of the genus Cu- lex were caught in 60% of the counties. Divan- dere and Saqez have the highest abundance and are considered as high-risk areas for this genus. Species of Culiseta were identified in more than 50% of the study areas, which was abun- dant in the central parts of the province. In gen- eral, the abundance of Culiseta was higher than that of Culex and Anopheles (Fig. 2). Inverse Distance Weighted (IDW) analysis The results of IDW for Anopheles species in the study area showed that there was a very extensive hotspot in the west of Kurdistan Province. High density of Anopheles species in these areas increase the risk of malaria trans- mission in case of detecting vector infection. These high-risk areas locate in three counties bordering Iraq. Two small hotspots were de- termined in Saqez for the genus Culex, which is limited to the county itself, and is slightly extended to the north of the province. On the one hand, the genus Culiseta has a medium hotspot, which exists in the south of the province and in the county of Sanandaj. Based on this analysis, the high-risk areas of the province are determined as for Anopheles in the west, for Culex in the north, and for the Culiseta in the south of the province (Fig. 3). Alpha diversity indices Based on the calculation for the Simpson index, the maximum biodiversity was observed in Sarvabad and the minimum in Bijar. The max- imum biodiversity, based on Shannon-Weiner index, was determined in Baneh and mini- mum in Bijar. Shannon-Weiner Evenness in- dex showed maximum and minimum evenness of species distribution in Qorveh and Bijar, re- spectively. For Hill 1 and Hill 2 index the max- http://jad.tums.ac.ir/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 354 http://jad.tums.ac.ir Published Online: Dec 31, 2022 imum and minimum calculated in Baneh and Bijar respectively. The amount of species rich- ness, based on Margalef’s index, was maximum in Baneh and minimum in Bijar. Based on Men- hinick’s index, the maximum and minimum spe- cies richness was reported in Kamyaran and Bijar respectively. The statistical analyzes showed that there is a significant difference between the counties in all calculated indices (Table 3). Table 1. Geographical coordinates of mosquito larval collecting sites, Kurdistan Province, summer, 2019 County Location Latitude Longitude County Location Latitude Longitude Sananadaj Sanandaj 35.320776N 46.967762E Bijar Gharatoreh 35.8070749N 47.454365E Tavrivar 35.128055N 46.983173 E Sarabghamish 35.329604N 47.047870E Nadri 35.7606463N 47.529502E Kanimeshkan 35.237838N 46.921330 E Sadeghabad 35.6553119N 47.169327E Babariz 35.364240N 47.065193E Khorkhoreh 35.9689153N 47.800325E Doiseh 35.455993N 46.895943E Salavatabad 35.9983444N 47.550961E Salavatabad 35.279508N 47.126181E Ghabasorkh 35.8569962N 47.408851E Hasanabad 35.260319N 46.968247E Mehrabad 35.9011767N 47.913803E Naran 35.154100N 47.067685E Sayedan 35.9409092N 47.725527E Baneh Nojneh 36.128394N 45.783805E Khosroabad 35.5182203N 47.623157E Aloot 36.025676N 45.571554E Najafabad 35.7932236N 47.238271E Savan 36.074269N 45.904967E Ghadimkhan 35.6248274N 47.642770E Ashtarabad 35.844582N 45.917856E Ghamchaghai 36.1659343N 47.625592E Showe 36.044065N 45.865924E Dehgolan Bolbanabad 35.1404632N 47.321070E Marivan Vilae 35.592742N 46.307395E Miraki 35.4350278N 47.290693E Chashniabad 35.653148N 46.031590E Sis 35.2065655N 47.279559E Darahtephi 35.537565N 46.099041E Bagahjan 35.3491741N 47.455589E Sarvabad Daranakhi 35.392268N 46.233966E Qorveh Mehdikhan 35.3456197N 47.649151E Ghalaji 35.358081N 46.282911E Farhadabad 35.4154735N 47.616394E Rezab 35.260049N 46.403304E Ghaleh 35.1335135N 47.803795E Saqez Pole- gheshlagh 36.091621N 46.340016E Majin 35.1711562N 47.946798E Siyahdare 36.139224N 45.984306E Bahraloo 35.195755N 48.121763E Kandalan 36.256530N 46.067004E Sarab 35.1344474N 47.790924E Khanemiran 36.127169N 46.590750E Kamyaran Bovanah 34.8655297N 46.956129E Cheshme- Saqez 36.262425N 46.300410E Shirvanah 34.7959729N 46.964364E Divandarreh Gavshalh 36.016072N 47.147437E Kamyaran 34.7966668N 46.940417E Kolah 35.789951N 47.052898E Alak 34.8033522N 46.843097E Hazarkaniain 35.769183N 46.813742E Aghajari 35.891579N 47.128461E Divandarreh 35.916419N 47.031293E http://jad.tums.ac.ir/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 355 http://jad.tums.ac.ir Published Online: Dec 31, 2022 Table 2. Number of species belonging to mosquitoes in the counties of Kurdistan Province, summer, 2019 Species County S a n a n d a j B a n e h M a r iv a n S a r v a b a d K a m y a r a n B ij a r S a q e z D iv a n d e r e Q o r v e h D e h g o la n Total (%) An. claviger 0 95 15 0 6 0 0 4 0 0 120 (2.05) An. maculipennis s.l. 3 136 269 52 1 0 0 52 0 0 513 (8.79) An. Superpictus s.l. 2 161 37 40 15 0 0 1 0 0 256 (4.39) Cs. longiareolata, 885 57 125 75 86 217 58 0 1 313 1817 (31.16) Cs. subochrea 1 0 8 0 0 0 0 0 0 0 9 (0.15) Cx. hortensis 116 78 41 29 1 0 1 0 4 43 313 (5.36) Cx.mimeticus 9 20 25 31 12 0 1 0 22 2 122 (2.09) Cx.perexiguus 0 0 96 39 0 0 5 7 0 0 147 (2.59) Cx.pipiens 20 56 80 13 29 0 440 96 27 8 769 (13.18) Cx.theileri 7 18 573 124 16 1 103 416 153 27 1438 (24.66) Cx. modestus 2 89 0 0 0 0 0 0 1 0 92 (1.57) Cx.territans 0 28 0 0 0 0 40 167 0 0 235 (4.03) Total 1045 738 1269 403 166 218 648 743 208 393 5831 (100) Fig. 1. Map of the mosquito larval collecting sites in Kurdistan Province, 2019 http://jad.tums.ac.ir/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 356 http://jad.tums.ac.ir Published Online: Dec 31, 2022 Fig. 2. Distribution of the mosquito genera belonging to Culicidae in counties of Kurdistan Province, summer, 2019 http://jad.tums.ac.ir/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 357 http://jad.tums.ac.ir Published Online: Dec 31, 2022 Fig. 3. Distribution of hot spots of the mosquito genera belonging to Culicidae in Kurdistan Province, summer, 2019 http://jad.tums.ac.ir/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 358 http://jad.tums.ac.ir Published Online: Dec 31, 2022 Table 3. The alpha diversity for mosquito species in Kurdistan Province, summer, 2019 County S y m p so n M e a n ( S D ) P -V a lu e * S h a n o o n M e a n ( S D ) P -V a lu e S h a n e v e n M e a n ( S D ) P -V a lu e H il 1 M e a n ( S D ) P -V a lu e * H il 2 M e a n ( S D ) P -V a lu e M a r g a le f M e a n ( S D ) P -V a lu e M n a h e n ic M e a n ( S D ) P -V a lu e Bane 0.36 (0.36) P < 0 .0 0 1 1.66 (0.44) P < 0 .0 0 1 0.45 (0.26) P < 0 .0 0 1 5.31 (2.84) P < 0 .0 0 1 4.40 (2.37) P < 0 .0 0 1 1.41 (0.81) P < 0 .0 0 1 0.65 (0.31) P < 0 .0 0 3 Bijar 1.00 (0.02) 0.01 (0.04) 0.007 (0.026) 1.01 (0.04) 0.93 (0.26) 0.03 (1.00) 0.35 (0.09) Dehgolan 0.71 (0.23) 0.53 (0.39) 0.84 (0.59) 1.80 (0.61) 3.20 (3.72) 0.53 (0.36) 0.39 (0.09) Divandareh 0.50 (0.15) 0.96 (0.26) 1.14 (0.43) 2.68 (0.70) 2.13 (0.69) 0.90 (0.21) 0.43 (0.07) Ghorveh 0.67 (0.23) 0.57 (0.41) 1.16 (0.71) 1.91 (0.84) 1.63 (0.64) 0.57 (0.37) 0.58 (0.13) Kamyaran 0.48 (0.30) 1.01 (0.63) 0.44 (0.18) 3.18 (1.90) 2.66 (1.67) 1.09 (0.57) 0.82 (0.28) Marivan 0.32 (0.08) 1.41 (0.29) 0.49 (0.11) 4.22 (1.11) 3.26 (0.78) 1.25 (0.18) 0.54 (0.17) Saghez 0.61 (0.11) 0.69 (0.20) 0.34 (0.08) 2.03 (0.42) 1.68 (0.31) 0.68 (0.28) 0.40 (0.17) Sanandaj 0.76 (0.15) 0.46 (0.23) 0.26 (0.18) 1.62 (0.33) 1.35 (0.25) 0.64 (.41) 0.47 (0.23) Sarvabad 0.28 (0.06) 1.42 (0.28) 0.58 (0.08) 4.24 (1.07) 3.58 (0.80) 1.13 (0.40) 0.68 (0.23) *Probability from Kruskal-Wallis test Discussion A total of 5831 mosquitoe larvae were col- lected, which included 12 species and three gen- era. The highest number of species caught be- longed to Cs. longiareolata (31%), and the low- est number belonged to Cs. subochrea. The highest number of Anopheles caught was from the border county of Baneh. This city has been one of the foci of malaria transmission (un- published information, Kurdistan Province Health Center). Based on the maps that, gen- erating using Arc GIS software, Baneh has been identified as one of the most important hot spots in terms of the existence of Anopheles species. Since, this county is an official border with Iraq, and on the other hand, is one of the well-known border markets in the whole coun- try, it annually receives many travelers from in- side and outside the country. Due to the pres- ence of potential vector of the disease in this area, presence of a positive case of malaria may lead to a local transmission. Therefore, this is- sue requires a special attention to borders con- trol and applying strict quarantine laws at the borders and entry points. On the other hand, rou- tine entomological studies are required in this county, as well. In a recent study, the highest http://jad.tums.ac.ir/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 359 http://jad.tums.ac.ir Published Online: Dec 31, 2022 number of Anopheles specimens caught was reported from Sarvabad City (24). The highest number of An. superpictus s.l. specimens in our study was reported from Baneh City; while, in a recent study from Sarvabad City (24). Anopheles superpictus s.l. and An. maculipennis s.l. have been reported as the predominant spe- cies in Hamedan and West Azerbaijan Prov- inces, respectively (31, 32). Moreover, in East Azarbaijan Province, An. maculipennis s.l. has been reported as the predominant species of the genus Anopheles (33). Over the past few decades, An. superpictus s.l., An. maculipennis s.l. and An. sacharovi have been reported from 29, 20 and 18 out of 31 provinces of the coun- try, respectively. Anopheles superpictus s.l. is more common in the western, and An. macu- lipennis s.l. and An. superpictus s.l. are more common in the north and northwest of the country (34). In this study, more than 50% of the caught species belonged to the genus Culex. Based on IDW analysis, two high-risk points for this genus were identified in north-western areas of the province. The highest number (46%) be- longed to Cx. theileri, and the lowest (2.9%) be- longed to Cx. modestus. Culex theileri mostly caught from Marivan City in the west of the province. This species has recently been iden- tified as a vector of Dirofilaria in the north of the country (35). In our study, seven species of the genus Culex were identified. In two other studies conducted in the province, four and five species of this genus were reported, respective- ly. In contrary to our study, Cx. modestus was not reported, and this species introduced for the first time in the province. In agreement to the result of our study, a study introduced Cx. theil- eri as the predominant species in the province (24). In another study, the highest number of caught species, in contrast to our study, was Cx. theileri (24, 36). In another study conducted in Sanandaj County, the predominant species caught in larval stage was Cx. pipiens, and in adult stage, like our study, was Cx. theileri (22). Like our study, in Hamedan, the predominant species was Cx. theileri (37), and in West Azerbaijan, Cx. pipiens was the predominant species (38). In East Azarbaijan Province, Cx. theileri species has been the most dominant species in the region (33). In the present study, two species of the genus Culiseta re- ported including Cs. longiareolata and Cs. sub- ochrea. Culiseta longiareolata had the highest abundance (31.16%) among all species. This species was predominant in Sanandaj City, which previous studies in this city confirm the same results (22). In a study conducted by Banafshi et al. (12), in selected areas of Kur- distan Province, was reported as the dominant species. In Bijar, excluding Cx. theileri spe- cies, all the collected species were Cs. longi- areolata, as the IDW analysis showed a hot spot for this species in this region. Similarly, Cs. longiareolata is the predominant species in Hamedan and West Azerbaijan Provinces (37, 38). In this study, no species belonging to the genus Aedes were reported by dipping tech- nique or using ovitraps. Although, in previous studies a species belonging to this genus, Ae. caspius, had been caught in different parts of the province as well as neighboring provinces (22, 24, 33, 36, 38). Calculation of biodiversity alpha indices showed that in most of the studied areas, the value of these indices is statistically different. This issue, in addition to the effect of confound- ing variables such as the manner and time of sampling, can be affected by different climates in the province. Different climates have creat- ed diverse ecological niches for the establish- ment of different species. The results of this study showed that areas with warmer climates had provided better conditions for higher bio- diversity such as Sarvabad City. In studies con- ducted in the north of the country, Mazanda- ran province, and northwest, East Azerbaijan Province, similar to our study, different values were reported for biodiversity indices in the study areas (39, 40). http://jad.tums.ac.ir/ J Arthropod-Borne Dis, Dec 2022, 16(4): 350–363 F Sharifi et al.: Biodiversity and … 360 http://jad.tums.ac.ir Published Online: Dec 31, 2022 Conclusion To sum up, the western counties of the prov- ince are considered the hotspots for anopheline mosquitos. Moreover, reporting malaria cases in the past, bordering of these counties with Iraq, and the high traffic of travelers have made these areas as potential foci of malaria trans- mission. Although, in our study no species be- longing to the genus Aedes were reported, pre- vious studies have identified species belong- ing to this genus. Because two border counties of the province are the source of many goods importing from southeast Asian countries, which are endemic to dengue fever, routine entomo- logical checks and rigorous quarantine inspec- tions on entry points are necessary to detect any suspicious vector entrance. Acknowledgements The authors are very grateful to Kurdistan University of Medical Sciences for its finan- cial support of this project. (Project code: IR. MUK.REC.1397/146). Ethical considerations The study was approved by the Ethical Committee of Kurdistan University of Medi- cal Sciences (IR.MUK.REC.1397/146). Conflict of interest statement The authors declare there is no conflict of interests. References 1. 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