4. Dr Kazemi RTL Iranian J Arthropod-Borne Dis, (2009), 3(1): 29-35 SH Moosa-Kazemi et al: Culicinae (Diptera: … 29 Original Article Culicinae (Diptera: Culicidae) Mosquitoes in Chabahar County, Sistan and Baluchistan Province, Southeastern Iran *SH Moosa-Kazemi, H Vatandoost, H Nikookar, M Fathian Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran (Received 12 July 2009; accepted 5 Sep 2009) Abstract Background: Mosquito-borne diseases are a major public health threat in Iran. The objective of this study was to de- termine the fauna of culicinae mosquitoes for future mosquito control programs. Methods: Three genera and eleven species of the subfamily Culicinae (Diptera: Culicidae) were collected by dipping technique and identified in Chabahar County, Sistan and Baluchistan Province, southeastern Iran, during January, February, and March 2007. Results: The collected species included: Aedes vexans (new occurrence record for the province), Culex arbieeni, Cx. bitaeniorhynchus, Cx. deserticola, Cx. hortensis, Cx. perexiguus, Cx. pipiens, Cx. pseudovishnui, Cx. pusillus, Cx. quinquefasciatus, Cx. sinaiticus, Cx. theileri, Cx. tritaeniorhynchus, Culiseta longiareolata, Ochlerotatus cabal- lus, Oc. caspius, and Uranotaenia unguiculata. Conclusion: Our observations indicate that, in South of Iran hot and wet climatic conditions support the persistence of culicinae mosquitoes. As our study, regular monitoring of culicinae mosquitoes in this area could be the most use- ful for mosquito control and mosquito-borne disease prevention. Keywords: Culicinae, mosquito, fauna, Chabahar, Iran. Introduction Mosquito-borne diseases such as ma- laria, West Nile, Sindbis viruses, Japanese en- cephalitis, and RiftValley fever are a major pub- lic health in some Asian countries (Yasoka and Levis 2007). Some of diseases such as ma- laria, West Nile, Sindbis as well as Dirofilaria immitis (dog heart worm) and D. repens (di- rofilariasis) had been previously reported in Iran (Naficy and Saidi 1970, Saidi et al. 1976, Azari-Hamidian et al. 2007). Malaria transmis- sion in Chabahar County occurs through the year, most of the malaria cases were observed from May to Nov almost in the age groups of 5-25 yr old (Moosa-Kazemi et al. 2006). Malaria is transmitted by five species: Ano- pheles culicifacies Giles s.l., An. dthali Pat- ton, An. fluviatilis James s.l., An. stephensi Liston and An. superpictus Grassi (Moosa- Kazemi et al. 2006). By now, 64 species, 3 subspecies, and 7 genera of mosquitoes have been reported in Iran (Azari-Hamidian 2007). Four genera of Culex, Culiseta, Ochlerotatus, Uranotaenia and 18 species of the subfamily Culicinae have been reported from Sistan and Baluchistan Province (Lotfi 1973, Zaim 1987). Zaim (1987) mentioned four genera and sixteen species of the subfamily Culicinae (Diptera: Cu- licidae) in Sistan and Baluchistan Province as follow: Culex arbieeni Salem, Cx. bitaenio- rhynchus Giles, Cx. deserticola Kirkpatrick, Cx. hortensis Ficalbi, Cx. perexiguus Theobald, Cx. pipiens Linnaeus, Cx. pusillus Macquart, Cx. pseudovishnui Colless Cx. quinquefascia- tus Say, Cx. sinaiticus Kirkpatrick, Cx. theil- *Corresponding author: Dr Seyed Hassan Moosa- Kazemi, E-mail: moosakazemi@sina.tums.ac.ir Iranian J Arthropod-Borne Dis, (2009), 3(1): 29-35 SH Moosa-Kazemi et al: Culicinae (Diptera: … 30 eri Theobald, Cx. tritaeniorhynchus Giles, Culiseta longiareolata Macquart, Ochlerotatus caballus Theobald, Oc. caspius Pallas, and Uranotaenia unguiculata Edwards. It is note- worthy that Culex mimeticus and Cx. terri- tans were found by Lotfi (1973) in this area. There is only one recent study on the mosquitoes in the county in relation to ma- laria and anophelines (Moosa-Kazemi et al. 2006). While it is necessary to study mosqui- toes in Chabahar, there are no more refer- ences on the culicine mosquitoes of the county. This paper presents the first report of the re- sults of the recent surveys of the culicinae mos- quitoes from samples taken from a range of larval breeding sites in which they occur in nature, and adult culicinae mosquitoes captured from various places. The aim was to obtain new data, which would be valuable to develop programmes for future planning of mosquito control in this area of southeastern Iran. Materials and Methods To better understand the Culicinae fauna of Chabahar County and gather information on their distribution and the kinds of larval breeding sites in which they occur in nature, a field trial study was performed from Jan to Mar 2007 in Sistan and Baluchistan Province. This study took place in five randomly selected rural villages in Chabahar County (57°14'N, 26°19'E). The County is bounded by Pakistan in the east, Hormozgan Province in the west, Kerman Province in the north, and Oman Sea in the south (Fig. 1). In 2007, the max and min mean monthly temperature was 33 °C and 5.5 °C in July and Jan, respectively. The total annual rainfall was 221 mm, the minof 3.5 mm in July and max of 64 mm in Apr. The mean an- nual relative humidity was 75%. The main eco- nomic activity of the people is husbandry of cow, goats and fishing. Based on available epi- demiological data from the Chabahar Health Center, some villages were short-listed and entomological surveys were carried out. On this basis, five villages with 1118 houses and 5513 population were selected. To study the mosquito fauna, sampling was car- ried out by dipping technique for collecting lar- vae and pyrethrum space spray collection (PSSC) for adult mosquitoes. The specimens were collected from five villages in the differ- ent topographical areas of the district includ- ing Baho-kalat, Beris, Dak-baho, Pollan, and Soldan. Pyrethroum space spray mosquitoes collection were conducted between 05.00 and 08.00 h monthly in eight fixed animal and hu- man shelter randomly on each villages before spraying, all the eaves, windows, doors and other exit points were closed and cloth sheets were spread on the floor. Pyrethrum (0.2% in kerosene) was sprayed using a pressurized hand sprayer. After spraying, the room was kept closed for 15 min and the knocked-down mos- quitoes were then collected from the floor sheet with forceps and placed in Petri-dishes lined with moist cotton. All collected mosquitoes were pinned and then were identified using the keys of Zaim and Cranston (1986), Harbach (1988), and Darsie and Samanidou-Voyadjoglou (1997). Mosquito name abbreviations were cited based on Reinert (2001). Larval habitats present within 500 m around each village were studied for mos- quito larvae two times per month. Five ran- domly selected larval breeding places lo- cated within a 500-m radius of the study site were also included and formed the sampling unit for mosquito larvae. All larvae were col- lected from natural habitats, generally feed- ing with seepage water, such as swamps, seep- ages, streams, river banks, drying river beds, pools, and grasslands. Samples were taken at intervals along the edge of each larval breeding place using a standard mosquito dipper (350 ml) depending on the size of the habitat. The larvae for each habitat were placed separately in whirl paks and transported to the laboratory where they were sorted by ge- Iranian J Arthropod-Borne Dis, (2009), 3(1): 29-35 SH Moosa-Kazemi et al: Culicinae (Diptera: … 31 nus and instar counted and recorded. The pu- pae were reared to adults in emergent cages and identified to species by morphological char- acters. Because we did not have enough emer- gent cages to separate the pupae from each habitat by date of collection, samples for each habitat type were pooled together in each site irrespective of date of collection. The larvae were preserved in lactophenol and the microscopic slides of the preserved larvae were prepared using Berlese medium. The culicinae larvae along with the description of their breeding sites were sent to the School of Public Health, Tehran University of Medical Sciences, where the author identified using the aforementioned keys. Results The number of the culicine mosquito species at the various sites of Chabahar is shown in Table 1. A total of 3824 adults and 5031 larvae of the culicine mosquitoes were collected, and were recognized including; Aedes vexans Meigen (indicate new occurrence re- cords for the province), Culex arbieeni, Cx. bitaeniorhynchus, Cx. deserticola, Cx. hort- ensis, Cx. perexiguus, Cx. pipiens, Cx. pseu- dovishnui, Cx. pusillus, Cx. quinquefascia- tus, Cx. sinaiticus, Cx. theileri, Cx. tritaenio- rhynchus, Culiseta longiareolata, Ochlero- tatus caballus, Oc. caspius s.l., and Uranotae- nia unguiculata. In PSSC collection Overall, Culiseta longiareolata predominated (22.8%), followed by Cx. perexiguus (16.8%), Cx. pipi- ens (14.2 %), and Cx. tritaeniorhynchus (8.8%). In the larval collection, of 5031 culicinae larvae collected, Culiseta longiareolata predominated (19.5%), followed by Cx. perexiguus (16.9%), Cx. pipiens (13.6 %), and Cx. tritaeniorhynchus (9.1%) respectively (Table 1). Seven species including Culiseta longiareolata, Ochlerotatus caspius s.l., Cx. perexiguus, Cx. pipiens, Cx. sinaiticus, Cx. theileri, and Cx. tritaeniorhyn- chus accounted for 85.1%, and 79.2% of the adult and larval collection respectively. The other species were less than 6% of the total. Culiseta longiareolata was the most frequent culicine mosquito collected at Chabahar, with total of 982, and 875 specimens, by larval col- lection and PSSC, respectively. Aedes vexans is recorded for the first time in Chabahar Cou- nty and Sistan and Baluchistan Province. Num- ber and localities of the larvae of culicinae mos- quitoes collected in this survey are shown in Table 2. Uranotaenia unguiculata, Oc. cabal- lus, Cx. quinquefasciatus, and Cx. arbieeni were collected only in the larval stage. Number and localities of the adult of the culicine mosqui- toes collected in this survey are shown in Table 3. Culiseta longiareolata, Cx. perexiguus, and Cx. pipiens were the most prevalent species respectively, and collected in almost all of the villages (Table 2 and 3). The highest numbers of culicines were collected in the Dak-Baho (1206 larvae and 717 adult specimens), and the low- est in the Beris (965 larvae and 627 adult speci- mens) (Table 2 and 3). Ochlerotatus caballus, Cx. quinquefasciatus, and Cx. arbieeni were collected from larval collection and other species from both SPSS and Larval (Table 2 and 3). Discussion Five genera and 17 species of culicine mosquitoes were collected and identified from Chabahar County in Sistan and Baluchistan Province, including the first record of Ae. vexans in the province. From 18 species of culicinae mosquitoes in Sistan and Baluchistan Province (Lotfi 1973, Zaim 1987), 16 species, which had already been recorded, were also collected in this investiga- tion. Two species Culex mimeticus and Cx ter- ritans are not recorded in this study. However, the occurrence of Aedes vexans in the province was not mentioned before (Zaim 1987), although some references counted Aedes vexans as the more common species of Guilan and Isfahan Provinces (Azari-Hamidian et al. 2002, Mousa- kazemi et al. 2000). Iranian J Arthropod-Borne Dis, (2009), 3(1): 29-35 SH Moosa-Kazemi et al: Culicinae (Diptera: … 32 Fig. 1. Map of Iran indicating the location of the study area in Chabahar County (Ch) situated in the south of Sistan and Baluchistan Province (S and B) Table 1. Number and prevalence of adult and larvae culicinae mosquitoes collected by pyrethrum space spray and larval collections in Chabahar County, Sistan and Baluchistan Province during January, February, and March 2007 Larvae Adult Species No Percentage No Percentage Aedes vexans 120 2.4 61 1.6 Culex arbieeni 15 0.36 - - Cx. bitaeniorhynchus 30 0.59 49 1.3 Cx. deserticola 120 2.4 137 3.6 Cx. hortensis 65 1.3 111 2.9 Cx. perexiguus 851 16.9 643 16.8 Cx. pipiens 685 13.6 543 14.2 Cx. pseudovishnui 120 2.4 5 0.13 Cx. pusillus 206 4.1 202 5.3 Cx. quinquefasciatus 146 2.9 - - Cx. sinaiticus 267 5.3 291 7.6 Cx. theileri 418 8.3 291 7.6 Cx. tritaeniorhynchus 458 9.1 337 8.8 Culiseta longiareolata 982 19.5 875 22.8 Ochlerotatus caspius s.l. 327 6.5 279 7.3 Oc. caballus 156 3.1 0 - Uranotaenia unguiculata 65 1.3 - - Total 5031 100 3824 100 Iranian J Arthropod-Borne Dis, (2009), 3(1): 29-35 SH Moosa-Kazemi et al: Culicinae (Diptera: … 33 Table 2. Number and locality of the culicine mosquito larvae, Chabahar County, Sistan and Baluchistan Province, January–March 2007 Villages Species Baho-Kalat Beris Dak-Baho Pollan Soldan No % Aedes vexans 40 - 51 29 - 120 2.4 Culex arbieeni - 1 9 - 5 15 0.36 Cx. bitaeniorhynchus 6 8 7 4 5 30 0.59 Cx. deserticola 29 24 36 31 - 120 2.4 Cx hortensis 14 13 15 11 12 65 1.3 Cx. perexiguus 243 84 192 204 128 851 16.9 Cx. pipiens 178 - 201 217 89 685 13.6 Cx. pseudovishnui 29 14 28 32 17 120 2.4 Cx. pusillus 40 37 39 51 39 206 4.1 Cx. quinquefasciatus 29 68 - 49 - 146 2.9 Cx. sinaiticus - 94 - - 173 267 5.3 Cx. theileri 124 101 98 - 95 418 8.3 Cx. tritaeniorhynchus 94 83 101 54 126 458 9.1 Culiseta longiareolata 209 304 272 101 96 982 19.5 Ochlerotatus caspius s.l. 69 71 78 63 46 327 6.5 Oc. caballus - 63 48 - 45 156 3.1 Uranotaenia unguiculata - - 31 16 18 65 1.3 Total 1104 965 1206 862 894 5031 100 Table 3. Number and locality of the adult culicine mosquitoes collected by Pyrethrum space spray, Chabahar County, Sistan and Baluchistan Province, January–March 2007 Villages Species Baho-Kalat Beris Dak-Baho Pollan Soldan No % Aedes vexans - 16 21 11 13 61 1.6 Cx.bitaeniorhynchus 13 8 - 19 9 49 1.3 Cx. deserticola 45 48 44 - - 137 3.6 Cx. hortensis - - 63 34 14 111 2.9 Cx. perexiguus 151 116 164 183 29 643 16.8 Cx. pipiens 89 96 101 76 181 543 14.2 Cx. pseudovishnui - - 5 - - 5 0.13 Cx. pusillus 69 - - 121 12 202 5.3 Cx. sinaiticus 81 94 - 46 70 291 7.6 Cx. theileri 103 - 48 82 58 291 7.6 Cx. tritaeniorhynchus 59 - 15 142 121 337 8.8 Culiseta longiareolata 167 172 219 153 164 875 22.8 Ochlerotatus caspius s.l. 38 77 37 70 57 279 7.3 Total 815 627 717 937 728 3824 100 In this study, two species of the tribe Aedini including; Ochlerotatus caspius s.l. and Oc. caballus were recorded in Chabahar County. These species were differentiated from each other in larval stage (Zaim and Cranston 1986). There is no information about the Oc. caspius sibling species (A or B) in the coun- try (Azari-Hamidian 2007). Culex mimeticus and Cx. territans had been previously repoted by Lotfi (1973) in Sistan and Baluchistan, whereas in this study have not been found. Culex pipiens was one of the most fre- quent culicine mosquitoes collected at Chaba- har, with total of 573 and 685 specimens, by PSSC and larval collection respectively. Culex Iranian J Arthropod-Borne Dis, (2009), 3(1): 29-35 SH Moosa-Kazemi et al: Culicinae (Diptera: … 34 pipiens can be separated from Cx. torrentium, and Cx. vegans by Harbach’s (1988) key us- ing seta 1-III-V, seta 1-M, seta 1-X, seta 1-C, and some other characters. Based on this key, only Cx. pipiens was identified. One species of Culiseta, Cs. longiareo- lata was found in this investigation. Results of this survey are the same of finding by previ- ous study in Sistan and Baluchistan Province (Zaim 1987). In this study were collected other cu- licine mosquitoes such as, Culex. theileri, and Cx. Tritaeniorhynchus. This species are po- tential vectors of human and domesticated ani- mal pathogens (Horsfall 1955, Harbach 1988). The species Cx theileri was one of the culicine mosquitoe collected at Chabahar, with total of 291, and 418 specimens by means of PSSC and larval collection, respectively. This species is the more prevalent species at higher altitudes and in rural areas of Isfahan (Mousa- kazemi et al. 2000), Zanjan (Ghavami and Ladonni 2005) and East Azerbaijan Provinces (Abai et al. 2007). Observations made during the present study agree with those of Simsek (2004) who noted that Cx. theileri in Turkey prefers large natural larval habitats that are generally present in rural areas. In conclusion, regular monitoring of mos- quitoes in this area could be the most timely and sustainable way to follow up mosquito ecology and raise awareness about mosquito control and mosquito-borne disease prevention. Acknowledgements The authors are grateful to Dr S Azari- Hamidian, School of Health, Guilan Univer- sity of Medical Sciences, Rasht, for review- ing the article. We also would like to express our appreciation to the people of villages in Chabahar for their kind cooperation during the study, many thanks also dedicated for the efforts of the field staff of Iranshahr Health Research and Training Chabahar Health Cen- ters. This study was financially supported by Diseases Management Center, Iranian Ministry of Health and Medical Education, and the In- stitute of Public Health Research, Academic Pivot for Education and Research, Tehran Uni- versity of Medical Sciences. The authors de- clare that they have no conflicts of interest. 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