J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 69 http://jad.tums.ac.ir Published Online: March 31, 2021 Original Article Fauna and Larval Habitat Characteristics of Mosquitoes (Diptera: Culicidae) in Kashan County, Central Iran, 2019 Tahereh Sadat Asgarian 1 ; *Seyed Hassan Moosa-Kazemi 1 ; *Mohammad Mehdi Sedaghat 1 ; Rouhullah Dehghani 2 ; Mohammad Reza Yaghoobi-Ershadi 1 1 Department of Medical Entomology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2Social Determinants of Health Research Center, Department of Environment Health, School of Public Health, Kashan University of Medical Sciences, Kashan, Iran *Corresponding authors: Dr Seyed Hassan Moosa-Kazemi, E-mail: moosakazemi@tums.ac.ir, Dr Mohammad Mehdi Sedaghat, E-mail: sedaghmm@tums.ac.ir (Received 08 Feb 2020; accepted 24 Jan 2021) Abstract Background: Mosquitoes are responsible for spreading devastating parasites and pathogens causing some important infectious diseases. The present study was done to better understand and update the fauna of Culicidae and to find out the distribution and the type of their larval habitats in Kashan County. Methods: This study was done in four districts of Kashan County (Central, Qamasr, Niasar and Barzok). Mosquito lar- vae were collected from 23 active larval habitats using a standard 350ml capacity mosquito dipper from April to late December 2019. The collected larvae were transferred to containers containing lactophenol, and after two weeks indi- vidually mounted in Berlese's fluid on a microscope slide and identified to species by morphological characters and valid keys. Results: In this study, a total of 9789 larvae were collected from urban and rural areas in Kashan County. The identified genera were Anopheles, Culiseta and Culex. In this study larvae of An. turkhudi, Cx. perexiguus, Cx. mimeticus, Cx. deserticola and Cs. subochrea were collected for the first time from Kashan County. Conclusion: The results of this study indicate the presence and activity of different mosquito species in Kashan County that some of them are vectors of arbovirus and other vector-borne diseases. Keywords: Mosquito; Larvae; Surveillance Introduction Arthropod-borne diseases have always been a major health problem in different parts of Iran. Transmission of malaria, filariasis, enceph-alitis and other arboviral diseases by mosquitoes have made mosquitoes the most important arthro- pods in medicine and health (1-3). The mos- quitoes are from the order Diptera, suborder Nematocera and the family Culicidae (4). Sev- en arboviral diseases, two bacterial diseases, four filarial and two protozoal diseases which are assumed to be transmitted by mosquitoes are found in Iran (1, 5). One genus and spe- cies (Mansonia uniformis) was added to Iranian mosquito checklist, thus there are 70 species and 8 (or 12) genera depending the classifica- tion of the tribe Aedini (6). This family has two subfamilies, Anophelinae and Culicinae in Iran. Azari-Hamidian et al. (1) reviewed mosqui- toes of Iran and their medical and veterinary importance. There are diseases caused by mos- quito-borne pathogens in Isfahan Province in- cluding Avian pox (chickens, pigeons and tur- keys), West Nile fever (horses and humans), Anthrax (sheep, cattle and goats), mosquito- borne filariases (camels) and bird malaria (tur- keys, hens/roosters, pigeons and migratory wa- terfowl). Because of their high adaptation to differ- ent habitats, mosquitoes are successful organ- isms and are found all over the world except Copyright © 2020 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. https://creativecommons.org/licenses/by-nc/4.0/ https://creativecommons.org/licenses/by-nc/4.0/ J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 70 http://jad.tums.ac.ir Published Online: March 31, 2021 Antarctica. Mosquito larvae are found in a range of habitats, including natural and artificial hab- itats, with temporary or permanent, stagnant or flowing waters, contaminated or clean waters, with or without vegetation, and even in the smallest places where water accumulates in it, such as pots, used tires and animal footprints (2, 3). Based on literature, In Isfahan Province, ten species of Anopheles: An. algeriensis, An. clavi- ger, An. dthali, An. maculipennis, An. marteri, An. messeae, An. multicolor, An. sacharovi, An. superpictus, and An. turkhudi have been report- ed (7-15). An. messeae was recorded based on egg chorion pattern (9). Zaim (10), Saebi (11) and Mousa-Kazemi et al. (12) reported the Anopheles sacharovi and An. maculipennis of the Maculipennis Group, based on morpholog- ical characters. The recent group species was also being verified later by PCR technique (13). Ladonni et al. (14) reported three species of An. dthali, An. marteri, and An. turkhudi to the fauna of the Isfahan province. Seven spe- cies of Culex have been reported in Isfahan Province including: Cx. hortensis, Cx. mimeti- cus, Cx. modestus, Cx. perexiguus, Cx. pipiens, Cx. territans, and Cx. theileri (10, 12, 16, 17). Culiseta longiareolata, Cs. annulata and Cs. subochrea are cited in the province (10, 12, 17). Three species of Ochlerotatus caspius s.l., Oc. pulcritarsis, and Aedes vexans have been re- ported in the province (10, 12, 18). Zaim (18) reported Uranotaenia unguiculata. Based on Ladonni et al. (14) there are at least 24 spe- cies and six genera in the province. Little information is available about mosqui- toes in Kashan County. Zaim (10) mentioned some aspects of mosquito larvae and adult ecol- ogy in Kashan County and relieved that the oc- currences of 14 species in this area. Doroud- gar et al. (15) studied the epidemiology of ma- laria in Kashan. This limited information are not sufficient as the basis to prevent possible epidemics of mosquito borne diseases. Because of the importance of mosquitoes in medicine and health and disturbances caused by bites, the special location of Kashan County, climate changes in the county, as well as the construc- tion of garden of birds in Qamsar that have led to the entry of birds from 17 different coun- tries into the area, therefore, fauna and ecolog- ical study and obtain data on larval habitats, species composition, active season, and activ- ity peak of mosquito larvae in Kashan County are very important in integrated vector manage- ment. An entomology check program was felt after more than three decades in the county and a specialized study was done to better under- stand and update the fauna of Culicidae in Kashan County and to find out the distribu- tion and the type of their larval habitats in the region, which can lead to preparation of basic information in the country and region and if necessary, be effective in adopting the right strategy of integrated vector management. Materials and Methods Kashan County is located in the north of Isfahan Province, between Karkas Mountains and the central desert of Iran. This county is bounded on the north by the central desert and the city of Qom, on the east by the desert plain, on the south by the towns of Borkhar and Mey- meh and Natanz and on the west by the city of Mahallat. Kashan County has four districts (Cen- tral, Qamsar, Niasar and Barzok) and seven cit- ies (Kashan, Meshkat, Qamsar, Niasar, Jow- shaqan, Kamoo and Barzok). This is a faunis- tic and descriptive-sectional study that was done in four districts of Kashan County including Central District (Kashan and Meshkat cities), Qamasr District (Qamasr, Kamoo and Jow- shaqan cities), Niasar District (Niasser City) and Barzok District (Barzok City). The climate of the county varies depending on ups and downs. The uplands are cold, foots are temperate and plains, and especially on the margins of the desert are tropical. This county has a special position due to located on a large chain of roads in the country, located on the north-south transit route and located on the main communication route between eastern, western, northern and southern cities and provinces (19). In this study, larval habitats were visited in 77 urban and rural areas of the county and 23 J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 71 http://jad.tums.ac.ir Published Online: March 31, 2021 active larval habitats were selected (Figs. 1, 2). Permanent or temporary, natural or artificial lar- vae habitats, with or without vegetation, slow flowing or still water and full or partial sun- light in urban or rural areas, visited and sam- pled monthly. Larvae and pupae of the mos- quitoes were collected using a standard 350ml capacity mosquito dipper from April to late De- cember 2019. Twenty dips were taken in each larval habitat. Sampling were always done by the same individual in the morning (08:00– 12:00h) or afternoon (15:00–18:00h). Larvae from small water bodies collected by eye drop- per. The collected larvae were transferred to petri dish containing lactophenol and the date, collection site and habitat type of the larvae were recorded with special code on the contain- ers and in the relevant forms and transferred to medical entomology laboratory of Tehran University of Medical Sciences (TUMS). Af- ter that the larvae in lactophenol were trans- parent, each larva was individually mounted in Berlese's fluid on a microscope slide and identi- fied to species by morphological characters and valid keys (20-22). Some mosquitoe specimens were deposited in the museum of medical en- tomology, TUMS. Results In this study, a total of 9789 larvae were collected from urban and rural areas of four districts of Central, Qamsar, Niasar and Bar- zok in Kashan County and were identified based on valid diagnostic keys at species lev- el. The identified genera was Anopheles 772 (7.89%), Culiseta 1706 (17.42%) and Culex 7311 (74.69%) which included 13 species. Cu- lex pipiens and Cx. theileri had the highest abun- dance with 3658 (37.36%) and 2555 (26.10%), respectively (Table 1). Ten larval habitats were artificial and 13 were natural. 45.8% of larval habitats were in urban areas and 54.2% in rural areas. Among the larval habitats in urban areas, Kashan had the highest and Meshkat (Central District) the least abundance of larvae. In the rural areas, the larvae habitats of Khoncheh (from Central District) and Qazaan (from Qamsar District) had the highest and lowest abundance of mos- quito larvae, respectively. In terms of mosqui- to fauna diversity, the Central District (Kashan and Meshkat cities) with 3 species (Cx. pipiens, Cx. theileri and Cs. longiareolata) had the least species diversity and districts of Barzok and Niasar with 11 species had the highest mos- quito larvae fauna diversity. In terms of time distribution, the highest number of larvae was in August and the lowest number of larvae was in December. In this study, Cx. pipiens, Cx. theileri, Cs. longiareolata and An. super- pictus s.l. showed the highest time dispersion. All Anopheles species in this study were col- lected from natural habitats. Among the Cu- licinae species, Cx. pipiens, Cx. theileri and Cs. longiareolata were collected from natural and artificial habitats and the rest of the spe- cies collected only from natural larval habi- tats. Larvae of An. superpictus s.l. and An. maculipennis s.l. were collected from perma- nent and temporary habitats and larvae An. claviger and An. turkhudi were found from a temporary habitat. The larval habitat charac- teristics of the species has been listed in Table 2. In view of the simultaneous presence of different larval species in larval habitats, Cx. theileri was collected from 23 habitats with all others larvae. This species had the highest distribution in Kashan County. Culiseta longi- areolata was collected from 20 larval habi- tats. Anopheles superpictus s.l. was collected from 10 larval sites in four districts of the coun- ty and had the highest dispersal among anophe- line species. Species of An. claviger, An. turkhudi, Cs. annulata and Cs. subochrea each with a larval habitat had the least distribution in Kashan County (Table 3). Table 4 shows the species caught and coordinates of the col- lection sites of mosquito larvae in Kashan County. J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 72 http://jad.tums.ac.ir Published Online: March 31, 2021 Table 1. Abundance of larvae of the mosquito species collected in Kashan County, central Iran, 2019 Species Districts of Kashan County Central Qamsar Niasar Barzok Total No. % No. % No. % No. % No. % An. superpictus s.l. 159 3.21 453 14.94 10 1.08 57 6.44 679 6.93 An. maculipennis s.l. 0 0 54 1.78 15 1.63 22 2.49 91 0.93 An. claviger 0 0 0 0 1 0.10 0 0 1 0.01 An. turkhudi 0 0 0 0 0 0 1 0.11 1 0.01 Cx. pipiens 2651 53.56 733 24.17 37 4 237 26.78 3658 37.36 Cx. theileri 1028 20.8 1233 40.67 176 19.07 118 13.33 2555 26.10 Cx. deserticola 175 3.53 105 3.46 68 7.37 92 10.40 440 4.5 Cx. perexiguus 205 4.14 29 0.96 71 7.70 31 3.50 336 3.43 Cx. hortensis 150 3.03 31 1.02 40 4.33 33 3.73 254 2.60 Cx. mimeticus 15 0.30 39 1.29 0 0 14 1.58 68 0.70 Cs. longiareolata 566 11.43 355 11.7 460 49.84 280 31.64 1661 16.96 Cs. annulata 0 0 0 0 43 4.66 0 0 43 0.44 Cs. subochrea 0 0 0 0 2 0.22 0 0 2 0.02 Total 4949 100 3032 100 923 100 885 100 9789 100 Table 2. Characteristics of larval habitats and percentage of mosquito larvae in Kashan County, central Iran, 2019 Percentage of Culicidae larvae (%) Characteristic Variables A n . su p e rp ic tu s s. l . A n . m a c u li p e n n is s . l. A n . c la v ig e r A n . tu rk h u d C x . p ip ie n s C x .d e se rt ic o la C x .h o rt e n si s C x . p e re x ig u u s C x . m im e ti c u s C x . th e il e ri C s. l o n g ia re o la ta C s. a n n u la ta C s. s u b o c h re a Permanence Permanent 90 33.3 0 0 17.6 75 60 62.5 66.7 39.1 40 0 0 Temporary 10 66.7 100 100 82.4 25 40 37.5 33.3 60.1 60 100 100 Water current Slow flowing 70 33.3 0 0 0 75 60 62.5 66.7 34.8 35 0 0 Still 30 66.7 100 100 100 25 40 37.5 33.3 65.2 65 100 100 Intensity of light Full sunlight 80 33.3 0 0 23.5 75 40 75 66.7 73.9 85 0 0 Partial sunlight 20 66.7 100 100 76.5 25 60 25 33.3 26.1 15 100 100 Turbidity Clear 90 66.7 100 0 52.9 87.5 80 75 66.7 87 85 100 100 Turbid 10 33.3 0 100 47.1 12.5 20 25 33.3 13 15 0 0 Vegetation With vegetation 90 100 100 100 58.8 100 100 87.5 100 56.52 10 100 100 Without vegetation 10 0 0 0 41.2 0 0 12.5 0 43.48 90 0 0 Substrate type Mud 20 66.7 100 100 76.5 25 40 37.5 33.3 17.4 60 100 100 Sand 80 33.3 0 0 23.5 75 60 62.5 66.7 82.6 40 0 0 Habitat type Natural 100 100 100 100 11.8 100 100 100 100 56.6 50 100 100 Artificial 0 0 0 0 88.2 0 0 0 0 43.4 50 0 0 J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 73 http://jad.tums.ac.ir Published Online: March 31, 2021 Table 3. Number of simultaneous presence of mosquito larvae in active larval habitats in Kashan County, central Iran, 2019 Species N o . o f L a r v a l h a b it a t A n . su p e rp ic tu s s. l . A n . m a c u li p e n n is s . l. A n . c la v ig e r A n . tu rk h u d C x . p ip ie n s C x .d e se rt ic o la C x .h o rt e n si s C x . p e re x ig u u s C x . m im e ti c u s C x . th e il e ri C s. l o n g ia re o la ta C s. a n n u la ta C s. s u b o c re a An. superpictus s. l. 10 * 2 - 1 7 6 4 5 3 10 10 - - An. maculipennis s. l. 3 2 * 1 1 3 3 3 3 2 3 3 1 1 An. claviger 1 - 1 * - 1 1 1 1 - 1 1 1 1 An. turkhudi 1 1 1 - * 1 1 1 1 1 1 1 - - Cx. pipiens 17 7 3 1 1 * 7 5 7 3 17 17 1 1 Cx. deserticola 8 6 3 1 1 7 * 5 6 3 8 8 1 1 Cx. hortensis 5 4 3 1 1 5 5 * 4 3 5 5 1 1 Cx. perexiguus 8 5 3 1 1 7 6 4 * 3 8 8 1 1 Cx. mimeticus 3 3 2 - 1 3 3 3 3 * 3 3 - - Cx. theileri 23 10 3 1 1 17 8 5 8 3 * 20 1 1 Cs. longiareolata 20 10 3 1 1 17 8 5 8 3 20 * 1 1 Cs. annulata 1 - 1 1 - 1 1 1 1 - 1 1 * 1 Cs. subochrea 1 - 1 1 - 1 1 1 1 - 1 1 1 * A B C D E F Fig. 1. Some of active larval habitats in the study areas of Kashan County, central Iran from April to late December 2019. A. Khoncheh (Permanent, Slow Flowing), B. Nabar (Natural and Permanent), C. Niasar (Temporary, Still), D. Barzok (Natural and Still), E. Kashan (Natural, Slow Flowing) and F. Kashan (Artificial and Still) J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 74 http://jad.tums.ac.ir Published Online: March 31, 2021 Table 4. Coordinates of the collection sites and mosquito larvae identified in Kashan County, central Iran, 2019 District Site of Sampling Longitude Latitude Mosquito larvae Identified Central Meshkat-Hamzer 51.25705 34.17636 Cx. theileri Central Meshkat 51.27324 34.18104 Cs. longiareolata, Cx. pipiens, Cx. theileri Central Taherabad 51.35628 34.0421 Cs. longiareolata, Cx. pipiens, Cx. theileri Central Ravand (Fish Farming) 51.37237 34.02696 Cs. longiareolata, Cx. pipiens, Cx. theileri Central Kashan University of Medical Science 51.41 34.01705 Cs. longiareolata, Cx. pipiens, Cx. theileri Central Wastewater treatment 51.39998 34.04713 Cs. longiareolata, Cx. pipiens, Cx. theileri Central Khoncheh 51.21894 33.98134 An. superpictus s. l., Cs. longiareolata, Cx. theileri, Cx. deserticola, Cx. hortensis, Cx. mimeticus, Cx. perexiguus, Cx. pipiens Central Jazeh 51.377 7 33.84543 Cs. longiareolata, Cx. pipiens, Cx. theileri Qamsar Moslemabad 51.50039 33.78592 An. superpictus s. l., Cs. longiareolata, Cx. theileri Qamsar Road of Javinan 51.48367 33.74182 An. superpictus s. l., Cs. longiareolata, Cx. deserticola, Cx. hortensis, Cx. pipiens, Cx. theileri Qamsar Farfahan- Wastewater 51.42875 33.77983 Cs. longiareolata, Cx. pipiens, Cx. theileri Qamsar Farfahan-River 51.43248 33.75867 An. maculipennis s. l., An. superpictus s. l., Cs. longi- areolata, Cx. pipiens, Cx. theileri, Cx. deserticola, Cx. hortensis, Cx. mimeticus, Cx. perexiguus Qamsar Mazgan 51.40234 33.73779 Cx. theileri Qamsar Bonrood 51.42037 33.72226 An. superpictus s. l., Cs. Longiareolata, Cx. deserticola, Cx. pipiens, Cx. theileri Qamsar Qazaan 51.39733 33.7058 Cs. longiareolata, Cx. deserticola, Cx. perexiguus, Cx. theileri Qamsar Qohroud 51.42307 33.67773 An. superpictus s. l., Cs. longiareolata, Cx. deserticola, Cx. pipiens, Cx. theileri, Cx. perexiguus Niasar Kamal-al-Molk Spring 51.09115 34.0962 An. superpictus s. l., Cs. longiareolata, Cx. pipiens, Cx. theileri, Cx. perexiguus Niasar Kamal-al-Molk 51.11026 34.08385 Cs. longiareolata, Cx. pipiens, Cx. theileri Niasar Niasar 51.15015 33.97601 An. maculipennis s. l., An. claviger, Cs. annulata, Cs. subochrea, Cs. longiareolata, Cx. deserticola, Cx. hortensis, Cx. pipiens, Cx. theileri, Cx. perexiguus Niasar Nashalj 51.07791 33.97987 Cs. longiareolata, Cx. perexiguus, Cx. pipiens, Cx. theileri Barzok Nabar 51.20775 33.88567 An. superpictus s. l., Cs. longiareolata, Cx. theileri Barzok Barzok-Spring 51.22939 33.78756 An. maculipennis s. l., An. superpictus s. l., An. turkhudi, Cs. longiareolata, Cx. deserticola, Cx. hortensis, Cx. pipiens, Cx. mimeticus, Cx. theileri, Cx. perexiguus Barzok Barzok-Khavid 51.22525 33.79302 An. superpictus s. l., Cs. longiareolata, Cx. theileri J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 75 http://jad.tums.ac.ir Published Online: March 31, 2021 Fig. 2. The location of larva sampling places in Kashan County, central Iran, 2019 Discussion In this study, 13 species of mosquito larvae were collected and identified from three gen- era of Culicidae. Larva of An. turkhudi was col- lected for the first time from Kashan County, the city of Barzok. Also among the species of Culex, larvae, Cx. mimeticus and Cx. deserti- J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 76 http://jad.tums.ac.ir Published Online: March 31, 2021 cola were found from Kashan County for the first time. Of the three species of Culiseta iden- tified in this study, Cs. subochrea was identi- fied for the first time in the county and found from a temporary natural habitat associated with Cs. annulata larvae in the city of Niasar. Zaim (10) reported Cx. modestus from Mahmoudabad of Kouhpayeh. We did not catch this species, because in recent years land leveling, destruc- tion of lowlands and highlands, improvement of streams and cementation of streams have reduced larval habitats in different areas of Mahmoudabad (now is named Meshkat City). Zaim (10) also reported An. multicolor from Shurab area. In the new divisions of the coun- try, this area has been separated from Kashan County and is an area in Aran and Bidgol Coun- ty, therefore, it was not one of our study areas. Anopheles superpictus s.l. This Anopheles is distributed throughout the Iranian Plateau and the slopes of the Alborz Mountains and southern Zagros, as well as the coastal plains of the Caspian Sea and the Per- sian Gulf (23) and is one of the seven species of malaria vectors in Iran (20). This species was the most abundant and distributed species among Anopheles in Kashan County with 87.95% of the Anopheles collected. Its larvae were collect- ed from Qamasr, Khoncheh, Barzok, Qohroud, Javinan, Ka-malal-Molk, Moslem Abad and Nabar from June to November. Previous stud- ies have also reported larvae of this species from different regions of Kashan County (10, 15). Barghamadi et al. (24) reported the best lar- val habitat for this species in Kohkiluyeh and Boyer-Ahmad Province, natural habitats with permanent and clear water, still, non-vege-tated with mud core. Moosa-Kazemi et al. (12) re- ported its larval habitat primarily on rocky at shallow depths and secondarily on sandy and sunny rivers also shallow creeks with muddy bed. In a study in Ardabil, larvae of this spe- cies were reported from permanent and tem- porary habitats with submerged vegetation and from riverbeds (25). In the study done in Pol Dokhtar County from Lorestan Province, the larvae of this species was mostly found in nat- ural habitats of edge and bed of rivers with permanent, slow flowing and clear water with external or submerged vegetation (26). So- leimani-Ahmadi et al. (27) collected this spe- cies from larval breeding habitats including per- manent habitats in riverbeds mostly without vegetation and algae also still and clear water with full sunlight and sandy substrates. In Kashan County, the best larval breeding habi- tats for this species was the natural habitat of river edge with permanent and Slow flowing water and external or submerged vegetation. Anopheles maculipennis s.l. This species was included 11.79% of total Anophelinae species collected from Kashan County. It was found from temporary natural habitats in Niasar (July), in Barzok (Septem- ber and October) and a permanent natural hab- itat in Qamasr (July and August), and 100% of its larval habitats in Kashan County had low to high vegetation. Zaim (10) reported this species only once in July 1984 from Qamsar. Larvae of this species have been reported in swamps, marshes, natural meadows, and wa- ter leaks in rivers and springs and in sandy beds of rivers and stagnant waters. This spe- cies is one of the malaria vectors in Iran and is the main vector in the Caspian coast in northern Iran (28). Anopheles claviger It occupies a variety of larval habitats but usually prefers non-polluted, permanent or semi- permanent waters and it has been reported in almost all provinces of the country and is wide- ly distributed in the north and central parts of Iran (29). Waters that are warmed by sunlight, on the margins of rivers and canals in stag- nant water or with slow flowing in areas with shades of plants and trees, are suitable for lar- val period (30). It is blood sucking outdoor and transfers Tularemia disease (29). In Kashan County, this species was found from Niasar in a temporary larval habitat with stagnant, semi- sunny, clear water and high vegetation in Au- gust. In 1984, Zaim (10) collected these mos- quito larvae from a larval breeding place with running water next to a water stream in Mahmoudabad area in December. J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 77 http://jad.tums.ac.ir Published Online: March 31, 2021 Anopheles turkhudi This species has been reported from the south and some parts of central Iran (29, 31). In this study, the larvae of this mosquito were found from Kashan County for the first time from a temporary natural larval breeding place with vegetation also still and turbid water with par- tial sunlight and muddy substrates in Barzok in September. Ladonni et al. (14) reported this species from Fereydunshahr for the first time in Isfahan Province. Culex pipiens Its larval breeding place include both natu- ral and artificial habitats. Among the larvae col- lected in Kashan County, this species was most abundant and was collected from April to late December. Among the different Districts of Kashan County, the most larvae of this spe- cies belonged to the Central District and the city of Kashan so that, out of 3,645 larvae Cx. pipiens, 2020 were collected from urban and rural Kashan, such as Kashan wastewater treat- ment plant, stabilization pond of Kashan Uni- versity of Medical Sciences and waste water. In Iran, this species has been often reported from natural habitats (8). Zaim (10) found it from the natural habitats. Moosa-Kazemi et al. (30) and Azeri-Hamidian (32) collected it from man-made habitats, artificial habitats and rice fields. In Kashan County, the best larval habitat for this species was temporary larval breeding place with still and clear water, vegetation and par- tial sunlight. Many arbovirus diseases are trans- mitted to humans by Cx. pipiens, such as the Rift Valley fever, Japanese encephalitis, and the West Nile. This species is primarily an orni- thophilic species that feeds on both man and animals (33). Culex theileri This mosquito is found in some parts of the world and has been reported from all provinc- es of Iran (34-38). In Kashan County, this spe- cies had the highest spatial distribution and was collected from all larval habitats in cities and villages all of the time study from April to December. Its larval habitats are diverse in Kashan County. The best habitat for its larvae was the natural, permanent and temporary hab- itat, with still and clear water, and fullest sun- light with or without vegetation. Zaim (18) re- ported it from natural habitats, Moosa-Kazemi et al. (37) and Yaghoobi-Ershadi et al. (38) re- ported it from natural and artificial habitats. Cx. theileri is a major vector of the Rift Val- ley fever virus in South Africa (39) and it is involved in the transmission of Dirofilaria im- mitis nematode to humans. In Iran, third instar larvae of Cx. theileri were naturally infected with D. immitis and may be the main vector of this parasite (35). Culex theileri also trans- mits West Nile and Sindbis viruses in vitro (40). Culex hortensis This species is widely distributed in the mountainous regions of Iran and has been re- ported from at least 23 provinces (10, 18, 32 and 41) and in Kashan County was collected from permanent natural larval breeding places in Central District, Khoncheh village (August), natural and permanent larval breeding places in Qamsar (August), natural and temporary lar- val habitat in Niasar (July and August) and natural and temporary larval habitat of Barzok (the months of September and October). The best habitat for larvae of this species in Kashan County was permanent habitats with slow flow- ing and clear water, full sunlight with vegeta- tion. Zaim (10) reported this species from Dehnar, Qamsar, Qohroud, Barzok and Niasar in 1984. Culex hortensis has been introduced as a vector of avian malaria In vitro (42). Culex deserticola Larvae of this species were found in larval habitats with Cx. hortensis and is reported for the first time from Kashan County. Larvae were collected in Central District from the natural and permanent larval habitats of Khoncheh vil- lage in September and October, in Qamsar Dis- trict from natural and permanent habitats of Qamsar City and villages of Qohroud and Qazaan in July and September, in Niasar Dis- trict from a natural and temporary habitat in city of Niasar in June and July and in Barzok District from a temporary natural habitat from June to October. Its best larval habitats for this species in Kashan County was permanent hab- J Arthropod-Borne Dis, March 2021, 15(1): 69–81 TS Asgarian et al.: Fauna and … 78 http://jad.tums.ac.ir Published Online: March 31, 2021 itats with slow flowing and clear water with vegetation and full or partial sunlight. Culex perexiguus It is distributed in most provinces in Iran (18). In Kashan County, this species was found from permanent or temporary natural larval habitats in Qamasr, Niasar, Barzok and the vil- lages of Qohroud, Kamalal-Molk and Khonch- eh from June to October. The best habitat for larvae of this species in Kashan was perma- nent habitats with slow flowing water, full sun- light and with vegetation. Zaim (10) found Cx. perexiguus (as Cx. univittatus) in Mahmoudabad of Kouhpayeh. Culex mimeticus It is widely distributed in Iran and its lar- vae are found along the rocks and on the riv- erside of mountainous areas with abundant veg- etation (10). The best habitat for larvae of this species in Kashan County was permanent hab- itats with clear slow flowing water, full sun- light, and with vegetation, and was found from larval habitats in Khoncheh (July), Qamasr (Sep- tember) and Barzok (August, September and October). In this study, Cx. mimeticus larvae were reported from Kashan County for the first time. Zaim (10) found only adult of this species from Qohroud. Culiseta longiareolata This species is widely distributed in Iran. This mosquito has a variety of larval habitats, and is found in clear to polluted waters and in fresh and saline waters (10, 43). In Kashan County, this species was an active mosquito species and its larvae were collected from dif- ferent habitats throughout the study from April to late December. 93.2% of larval habitats of this species in Kashan County had no vegeta- tion and 100% of them had still water. Za- hirnia et al. (26) also collected 70% of the larvae of this species in Pol Dokhtar County from without vegetation habitats. It often is blood sucking from birds and rarely attacks humans. Although it feeds only on birds in North Africa, it feeds on large domestic ani- mals in Europe, including France. This spe- cies has medical importance. Adults are vec- tors of brucellosis, avian influenza virus, West Nile (44) as well as avian malaria (42). Its lar- vae are known as mosquito larvae predators (45). Culiseta annulata In Iran, larvae have been found from moun- tainous areas and the Alborz and Zagros Moun- tains. It has also been reported from Bushehr (46). Azari-Hamidian (43) studied the larval habitat characteristics and associated species in Guilan Province. In Kashan County, this mosquito Larvae was found from a temporary natural habitat, partial sunlight with high veg- etation, still and clear water in Niasar in June and July. This larval habitat was dried in mid- July. Larvae of this species were not found in the other studied areas. Zaim (10) reported lar- vae of this species in June from temporary lar- val habitats with still fresh water in Qohroud and from permanent larval habitats with sa- line flowing water from Mahmoudabad in Oc- tober and November. Azari-Hamidian (43) re- ported it from both natural and artificial habi- tats. Culiseta subochrea Culiseta subochrea is found in a number of provinces in Iran (47-49). In Kashan County, this species was found in a temporary larval habitat with still and clear water, partial sun- light and tall vegetation with Cs. annulata in Niasar. Paksa et al. (48) collected this species from stagnant and turbid water habitats with muddy substrate and shallow depth. Conclusion The results of this study indicate the pres- ence and activity of different mosquito spe- cies in Kashan County. Considering the dis- persion of An. superpictus s.l. in different dis- tricts of Kashan County, its abundance and since it's one of the seven malaria vectors in the country, a study is needed to identify its complex species. This study could be a basis for more comprehensive research about the im- portance of mosquitoes in medicine and health. We declare that we have no conflict of inter- est. 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