6. Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 37 Original Article Larval Habitat Characteristics of the Genus Anopheles (Diptera: Culicidae) and a Checklist of Mosquitoes in Guilan Province, Northern Iran S Azari-Hamidian School of Health, Guilan University of Medical Sciences, Rasht, Iran (Received 7 Aug 2010; accepted 9 Apr 2011) Abstract Background: Ecological data are important in the vector control management of mosquitoes. There is scattered pub- lished information about the larval habitat characteristics and ecology of the genus Anopheles (Diptera: Culicidae) in Iran and most of available data is in relation to malaria vectors in southern Iran. Methods: This cross sectional investigation was carried out to study the mosquito fauna and ecology in Guilan Province, northern Iran, during April–December 2000. Larvae were collected using the standard dipping technique. Larval habitat characteristics were recorded according to water situation (clear or turbid), vegetation, substrate type, sunlight situation, habitat situation (transient or permanent, running or stagnant), habitat type (natural or artificial), and water temperature. Results: In total, 1547 third- and fourth-instar larvae of Anopheles from 90 habitats were collected and morphologi- cally identified. Five species; Anopheles claviger, An.’hyrcanus’, An. maculipennis s.l., An. plumbeus, and An. su- perpictus were identified and respectively comprised 6.3%, 22.4%, 54.4%, 13.0%, and 3.9% of the samples. The mean and range temperatures of the larval habitat water were 19.6oC (n=14) (16–25oC), 22.6oC (n=53) (12–33oC), 23.8oC (n=52) (10–33oC), 11.5oC (n=12) (9–21oC), and 20.4oC (n=7) (12–26oC), respectively. There was a signifi- cant difference in the mean water temperatures (11.5–23.5oC) of the larval habitats of different species (P=0.000). Most of the genus larvae were collected from natural habitats (86.9%) such as river bed pools (46.4%) and rain pools (33.1%) with transient (98.3%), stagnant (99.5%) and clear (95.3%) water, with vegetation (69.9%), mud (42.0%) or gravel (39.7%) substrate in full sunlight (69.6%) or shaded (22.7%) area. A checklist of the province mosquitoes including 30 species and seven genera has been provided. Conclusion: The main larval habitats of the most abundant species, An.’hyrcanus’ and An. maculipennis s.l., in Guilan Province are: river bed pools, rain pools, and rice fields. Keywords: Anophelinae, Ecology, Iran, larvae Introduction According to the most recent classifica- tion of mosquitoes, the family Culicidae (Diptera) includes two subfamilies, 11 tribes, 113 genera, and 3531 species in the world fauna and the genus Anopheles Meigen in- cludes seven subgenera and at least 465 spe- cies (Harbach 2007). Certain species of Anopheles are involved in the transmission of various arboviral and filarial diseases to humans and domestic animals and/or are im- portant for their biting in different parts of the world, but the most important disease transmitted by them is malaria. About 70 Anopheles species are malaria vectors in which about 40 are important vectors (Ser- vice 1993). Sindbis virus was reported in Iran as well as West Nile virus in Iran and Guilan Prov- ince of the Caspian Sea littoral, northern Iran (Naficy and Saidi 1970, Saidi et al. 1976); Corresponding author: Dr Shahyad Azari-Hamidian, E-mail: azari@gums.ac.ir Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 38 however there is no information about their vectors in the country. The mosquito-borne filariae; Dirofilaria Railliet and Henry (diro- filariasis) and Setaria Viborg (setariasis) (Spirurida: Onchocercidae) have been re- ported in Iran and Guilan Province (Eslami 1997, Azari-Hamidian et al. 2007). Anophe- les maculipennis Meigen and Culex theileri Theobald are known vectors of Setaria labi- atopapillosa (Alessandrini) and Dirofilaria immitis (Leidy) respectively, in Ardebil Province, northwestern Iran (Azari-Hamidian et al. 2009). Malaria is the most important mos- quito-borne disease in Iran, especially in south- eastern areas, and seven species have been assumed to play role as malaria vectors: An. sacharovi Favre, An. maculipennis sensu lato (s.l.), An. fluviatilis James s.l., An. ste- phensi Liston, An. superpictus Grassi, An. dthali Patton, and An. culicifacies Giles s.l. (Edrissian 2006). Also, Zaim et al. (1993) re- ported An. pulcherrimus Theobald as a po- tential vector in southeastern Iran. Eshghy (1977) observed Plasmodium oocysts in An. multicolor Cambouliu, but sporozoites have not been detected in this species and it is not considered a vector in Iran. Recently, Djadid et al. (2009) reported An. hyrcanus (Pallas) as a potential vector of malaria based on nested Polymerase Chain Reaction (PCR) in Guilan Province. The most recent checklist of Iranian mos- quitoes includes 64 species and three sub- species belonging to seven genera (Azari- Hamidian 2007a). Almost half of the Iranian mosquito species (31 species) and seven genera have been previously recorded in Guilan Province (Zolotarev 1945, Dow 1953, Lotfi 1973, Danilov 1975, Saebi 1987, Zaim 1987b, Harbach 1988, Momeni et al. 1992, Azari- Hamidian et al. 2002b, Sedaghat et al. 2003, Gholizadeh et al. 2004, Gholizadeh et al. 2005). Despite of the importance of ecological data in the vector control management of mosquitoes, there is scattered published in- formation about the larval habitat character- istics and ecology of Anopheles in Iran and most of available data is in relation to ma- laria vectors in southern Iran. Marsh (1933) showed some larval breeding places of An. apoci Marsh larvae when described it for the first time in Iran. Macan (1950) studied the different aspects of some Anopheles species in northern and western Iran. Dow (1953) and Yaghoobi-Ershadi et al. (1986) stressed the larval breeding sites and associate spe- cies of some anophelines in the different ar- eas of the country and southern Iran, respec- tively. Manouchehri and Rohani (1975) studied the larval habitats of An. dthali in southern Iran. Zaini et al. (1975) and Manouchehri et al. (1976) mentioned the larval breeding place characteristics of An. stephensi in southern Iran. Eshghi et al. (1976) stressed some larval breeding places of An. fluviatilis s.l. in southern Iran. Eshghy (1977) noted the adult and larval ecology of An. multicolor. Zaim (1987a) studied the mosquito fauna of Kashan of Isfahan Province in central Iran with some notes on the larval breeding places and ecol- ogy of four anopheline species. Mousaka- zemi et al. (2000) studied the fauna and ecology of mosquitoes including An. macu- lipennis s.l. and An. superpictus in Zarrin- Shahr and Mobarakeh areas of Isfahan Prov- ince. Yaghoobi-Ershadi et al. (2001) stressed some information on the larval stage of An. sacharovi and An. superpictus in Ardebil Province of northwestern Iran. Azari-Hami- dian et al. (2002a) showed some larval breeding places of mosquitoes including anophelines in Rasht County of Guilan Province in northern Iran. Ghanbari et al. (2005) studied some physical and chemical factors of the larval breeding places of the eight species of Anopheles in Iranshahr, southeastern Iran. Investigation on the mosquito ecology and larval habitats is important in different aspects including source reduction in vector control through modifications of these habi- tats. This helps us to decrease, even if not very much, vector population and density Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 39 then vector-reservoir and vector-pathogen contacts, and biting nuisance. This may de- crease the transmission of different mos- quito-borne diseases and the burdens of these diseases. The present article is focused on some aspects of the larval ecology of Anopheles species, such as the larval habitat characteristics, water temperature, associated species, and provides a checklist of mosqui- toes in Guilan Province. Materials and Methods Study area Guilan Province in the Caspian Sea litto- ral of northern Iran, between Caspian Sea and Alborz Mountain range, has coastal, plain, foothill, and mountainous areas with an area of approximately 14700 square kilo- meters. The province with temperate climate and relatively warm-humid summer is located between 36o34'–38o27' N latitude and 48o34'– 50o36' E longitude and formally includes 16 counties; Amlash, Astaneh-e-Ashrafiyeh, Astara, Bandar-e-Anzali, Fuman (Fooman), Lahijan, Langrud (Langroud), Masal, Rasht, Rezvan- shahr, Rudbar, Rudsar (Roudsar), Shaft, Siahkal, Some’e Sara, and Talysh (Talesh). Guilan Province with about 1200 mm annual rainfall has the greatest rainfall in Iran and the main agricultural crop is rice. This prov- ince with vast deciduous forests of Hyrcania and temperate climate is a great location to breed mosquitoes. Specimen and data collection and analysis In this cross sectional investigation, in each county (in total 16) one fixed and three variable sites (including different topog- raphical areas) randomly selected and larval specimen collection was carried out for 15– 20 minutes during spring, summer, and au- tumn seasons in 2000. Thus, larval collection was carried out at least three times in fixed sites and 9 times in variable sites during three seasons in each county. Larvae were col- lected from natural breeding sites such as river edges, river bed pools, rain pools, marshes, grasslands, and tree holes and from artificial breeding sites such as rice fields, irrigation channels, wells, discarded tires, and buckets using the standard dipping tech- nique (350 mL dipper) and also by means of pipette (for small habitats) and bucket (for wells) (WHO 1975). Physical and biological characteristics of larval habitats including habitat situation (permanent or transient, stagnant or running), habitat type (natural or artificial), vegetation situation (with or with- out vegetation), substrate type, sunlight situation (full or partial sunlight or shaded), water situation (clear or turbid), and water temperature were recorded. The continuous variable of the temperatures of larval habi- tats was analyzed by One-Way ANOVA analysis using SPSS (Version 11.5 for win- dows, SPSS Inc., Chicago, IL) software. The third- and fourth-instar larvae and adult speci- mens were identified using the keys of Shah- gudian (1960), Zaim and Cranston (1986), and Harbach (1988). The mosquito name genera and subgenera abbreviations followed Reinert (2001). Taxonomic note Two close species Anopheles hyrcanus and An. pseudopictus Grassi of the Hyrcanus Group are identified based on the characters of adult in Guilan Province (Dow 1953, Azari-Hamidian et al. 2002b, 2003a, 2006), these species have not been identified in lar- val stage in this investigation and mentioned as An.’hyrcanus’ (An. hyrcanus group). Dar- sie and Samanidou-Voyadjoglou (1997) mentioned just one character of seta 2-C (in- ner clypeal seta) to distinguish the larvae of An. hyrcanus from those of An. pseudopic- tus. This seta is simple in An. pseudopictus and bears some short apical branches in An. hyrcanus. This character needs to be studied carefully in Iran. Seven species of An. maculipennis complex have been recorded in Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 40 the province based on morphological char- acters including egg pattern and PCR tech- nique (Dow 1953, Azari-Hamidian et al. 2002b, Sedaghat et al. 2003, Gholizadeh et al. 2004, Gholizadeh et al. 2005). Only An. sacharovi is reliably distinguishable from other species in adult and larval stage. In larvae, the mean number of seta 2 branches of the fourth and fifth abdominal segments in An. sacharovi is 36.8 whereas it is 16.5 for An. maculipennis s.l. (Doosti et al. 2006). It seems the use of PCR technique is the only reliable way to distinguish other species of the group, so the Maculipennis Group larvae cited as An. maculipennis s.l. herein. Results Larval habitat characteristics and ecology In these investigations, totally 6656 lar- vae including 1547 anopheline larvae (23.2%) and 5109 culicine larvae (76.8%) were collected from 127 larval breeding sites during 55 occasions. Among 127 larval breeding sites 14 (11.0%) contained only subfamily Anophelinae, 37 (29.2%) included only subfamily Culicinae, and 76 (59.8%) contained both subfamilies. In total, anopheline larvae were found in 90 breeding sites (70.8%) and culicine larvae in 113 ones (88.9%). In the present study, 1547 larvae of the genus Anopheles were collected during 11 occa- sions (20.0% of the total). Five species of Anopheles; An. claviger (Meigen) (6.3%), An.’hyrcanus’ (22.4%), An. maculipennis s.l. (54.4%), An. plumbeus Stephens (13.0%), and An. superpictus (3.9%) were found among the samples (Table 1). The associa- tion occasions of Anopheles species with other collected species are shown in Table 2. Most of the genus larvae were collected from natural habitats (86.9%) such as river bed pools (46.4%) and rain pools (33.1%) with transient (98.3%), stagnant (99.5%) and clear (95.3%) water, with vegetation (69.9%), mud (42.0%) or gravel (39.7%) substrate in full sunlight (69.6%) or shaded (22.7%) area (Table 3). The association percentages of Anopheles species with other collected spe- cies are shown in Table 4. There is a signifi- cant difference in the mean water tempera- tures of the larval habitats of different spe- cies (P= 0.000). Anopheles claviger Anopheles claviger larvae were collected during 11 occasions (20% of the total) from 14 larval breeding sites (11.0% of the total and 7.9% of the anopheline larval habitats) in April, May, June, and September 2000. The maximum and minimum water temperatures of larval habitats were 25o C and 16o C re- spectively, and mean temperature was 19.6o C (for 14 larval breeding sites) (SD= 3.0). Anopheles ’hyrcanus’ Anopheles ’hyrcanus’ larvae were col- lected during 33 occasions (60% of the total) from 53 larval breeding sites (41.7% of the total and 4.4% of the anopheline larval habitats) during April–November 2000. The maximum and minimum water temperatures of larval habitats were 33o C and 12o C respec- tively, and mean temperature was 22.9o C (for 53 larval breeding sites) (SD= 4.7). Anopheles maculipennis s.l. Anopheles maculipennis s.l. larvae were collected during 30 occasions (54.5% of the total) from 53 larval breeding sites (41.7% of the total and 3.5% of the anopheline larval habitats) during April–November 2000. The maximum and minimum water temperatures of larval habitats were 33oC and 10oC re- spectively, and mean temperature was 23.8o C (for 52 larval breeding sites) (SD= 4.0). Anopheles plumbeus Anopheles plumbeus larvae were col- lected during 9 occasions (16.3% of the to- tal) from 12 larval breeding sites (9.4% of the total and 3.5% of the anopheline larval Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 41 habitats) in October and November 2000. The maximum and minimum water temperatures of larval habitats were 21oC and 9o C respec- tively, and mean temperature was 11.5o C (for 12 larval breeding sites) (SD= 3.2). The mean temperature of the larval habitat water of this species showed significant difference from other species (P= 0.000). Anopheles superpictus Anopheles superpictus larvae were col- lected during four occasions (7.2% of the total) from 7 larval breeding sites (9.4% of the total and 3.5% of the anopheline larval habitats) in May, August, and October 2000. The maximum and minimum water tempera- tures of larval habitats were respectively 26o C and 12o C, and mean temperature was 20.4o C (for 7 larval breeding sites) (SD= 6.3). Checklist of the mosquitoes of Guilan Prov- ince The checklist of 30 species of mosquitoes in seven genera represented in Guilan Prov- ince is given below. The species which were recorded in the province by other authors and/or using PCR technique and were not found or identified in this investigation are shown by asterisks (*): Anopheles algeriensis Theobald*, An. atroparvus van Thiel*, An. claviger (Meigen), An. hyrcanus (Pallas), An. labranchiae Falleroni*, An. maculipennis Meigen, An. melanoon Hackett, An. messeae Falleroni, An. persiensis Linton, Sedaghat and Harbach*, An. plumbeus Stephens, An. pseudopictus Grassi, An. sacharovi Favre*, An. superpictus Grassi, Aedes vexans (Meigen), Coquillettidia richiardii (Ficalbi), Culex hortensis Ficalbi, Cx. mimeticus Noe, Cx. pipiens Linnaeus, Cx. territans Walker, Cx. torrentium Martini, Cx. theileri Theobald, Cx. tritaeniorhynchus Giles, Culiseta annulata (Schrank), Cs. longiareolata (Macquart), Cs. morsitans (Theobald), Cs. subochrea (Ed- wards)*, Ochlerotatus caspius (Pallas) s.l., Oc. echinus (Edwards), Oc. geniculatus (Oliv- ier), Uranotaenia unguiculata Edwards. Table 1. The composition and abundance of Anopheles larvae in Guilan Province, Iran, April– December 2000 Taxon n Percentage of family (%) Percentage of genus (%) An. claviger 97 1.46 6.27 An.’hyrcanus’ 346 5.20 22.37 An.maculipennis s.l. 841 12.63 54.36 An. plumbeus 202 3.03 13.06 An. superpictus 61 0.92 3.94 Total Anophelinae 1547 23.24 100 Total Culicidae 6656 - - Table 2. The association occasions of Anopheles larvae with different mosquito larvae in Guilan Province, Iran, April–December 2000 Species T otal occasion s A n . cla vig er A n .’hyrca n u s’ A n . m a cu lip en n is s.l. A n . su p erp ictu s A e. vexa n s C x. m im eticu s C x. p ip ien s C x. th eileri C x. trita en io rh yn ch u s C x. h orten sis C x. territa n s C s. lo n g ia reo lata C s. m o rsita n s C s. a n n u la ta O c. ech in u s O c. gen icu la tu s U r. u ng u icu la ta An. claviger 14 - 4 4 1 3 - 2 - 1 4 4 3 - 1 - - - An.’hyrcanus’ 53 4 - 39 4 6 12 13 7 31 4 5 1 1 2 - - 1 An. maculipennis s.l. 53 4 39 - 3 1 10 14 8 30 2 5 1 1 - - - - An. plumbeus 12 - - - - - - 1 - - - - - - - 4 1 - An. superpictus 7 1 4 3 - - 2 1 2 5 2 - 1 - - - - 1 Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 42 Table 3. The larval habitat characteristics and occurrence percentages of Anopheles larvae in Guilan Province, Iran, April–December 2000 Larval breeding site- characteristics and habitats A n . cla vig er (% ) A n .’h yrca n u s’ (% ) A n . m a cu lip en n is s.l. (% ) A n . p lu m beu s (% ) A n . su p erp ictu s (% ) A n o p h eles (% ) A) Habitat situation 1. Permanent 7.2 2.6 1.2 - - 1.7 2. Transient 92.8 97.4 98.8 100 100 98.3 3. Slow-running water 5.2 0.6 0.1 - - 0.5 4. Stagnant water 94.8 99.4 99.9 100 100 99.5 B) Vegetation situation 1. Without vegetation 33.0 14.7 16.8 100 65.6 30.1 2. With vegetation 67.0 85.3 83.2 - 34.4 69.9 2a) Emergent 78.9 51.4 30.2 - 66.7 39.8 2b) Floating - 0.6 0.4 - - 0.4 2c) Submerged 21.1 48.0 69.4 - 33.3 59.8 C) Substrate type 1. Mud 73.7 56.1 30.7 - 67.8 42.0 2. Sand 8.4 20.8 19.7 - - 18.3 3. Gravel 17.9 23.1 49.6 - 32.2 39.7 D) Water situation 1. Turbid - 1.7 8.0 - - 4.7 2. Clear 100 98.3 92.0 100 100 95.3 E) Sunlight situation 1. Full sunlight 22.7 71.7 89.4 - 88.5 69.6 2. Partial sunlight 11.3 21.7 4.1 - - 7.7 3. Shaded 66.0 6.6 6.5 100 11.5 22.7 F) Habitat type 1. Natural habitat 97.9 83.2 83.6 98.0 100 86.9 1a. River edge - - 0.1 - - 0.1 1b. River bed pool 28.4 37.2 67.0 - 31.1 46.4 1c. Stream edge 5.3 7.6 0.3 - - 2.2 1d. Grassland - 1.0 0.7 - 1.7 0.7 1e. Marsh - 13.2 - - - 2.8 1f. Rain pool 66.3 41.0 31.9 - 67.2 33.1 1g. Tree hole - - - 100 - 14.7 2. Artificial habitat 2.1 16.8 16.4 2.0 - 13.1 2a. Rice field - 79.4 61.6 - - 64.8 2b. Rice irrigation channel - 10.3 26.8 - - 21.3 2c. Well 100 - - - - 1.0 2d. Discarded concrete tube - - 5.1 - - 3.5 2e. Discarded tire - - - 100 - 2.0 2f. Water storage pool - 10.3 6.5 - - 7.4 Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 43 Table 4. The association percentages of Anopheles larvae with different mosquito species larvae in Guilan Province, Iran, April–December 2000 Species association Abundance (%) An. claviger Alone 19.59 Cx. hortensis, Cs. longiareolata 17.53 Cx. territans 12.37 Ae. vexans 9.28 An.’hyrcanus’, An. maculipennis s.l., Ae. vexans, Cx. pipiens 9.28 An. superpictus, Cx. hortensis, Cs. longiareolata 8.25 An.’hyrcanus’, An. maculipennis s.l. 7.22 Cx. hortensis, Cs. annulata, Cs. longiareolata 5.15 An. maculipennis s.l., Cx. territans, Cx. tritaeniorhynchus 4.12 Cx. hortensis 3.09 An.’hyrcanus’, Ae. vexans, Cx. territans 2.06 Cx. pipiens, Cx. territans 2.06 Total 100 An.’hyrcanus’ An. maculipennis s.l., Cx. tritaeniorhynchus 27.75 Ae. vexans, Cx. pipiens, Cs. annulata 10.98 An. maculipennis s.l., Cx. mimeticus 9.83 An. maculipennis s.l. 7.80 An. maculipennis s.l., Cx. mimeticus, Cx. tritaeniorhynchus 7.51 An. maculipennis s.l., Cx. pipiens, Cx. theileri, Cx. tritaeniorhynchus 3.18 An. maculipennis s.l., Cx. pipiens, Cx. theileri 2.90 Cx. tritaeniorhynchus 2.89 Alone 2.60 Cx. hortensis, Cx. theileri, Cx. tritaeniorhynchus 2.60 An. maculipennis s.l., Cx. theileri, Cx. tritaeniorhynchus 2.31 Ae. vexans, Cx. tritaeniorhynchus 2.02 An. claviger, Ae. vexans, Cx. territans 1.73 An. claviger, An. maculipennis s.l. 1.73 An. maculipennis s.l., Cx. territans 1.73 An. claviger, An. maculipennis s.l., Ae. vexans, Cx. pipiens 1.44 An. maculipennis s.l., Cx. hortensis, Cx. tritaeniorhynchus 1.44 An. maculipennis s.l., Cx. pipiens, Cx. theileri, Cx. tritaeniorhynchus, Cs. longiareolata 1.44 An. maculipennis s.l., Cx. pipiens, Cx. tritaeniorhynchus 1.16 An. maculipennis s.l., An. superpictus, Cx. mimeticus, Cx. tritaeniorhynchus 1.16 An. maculipennis s.l., Cx. pipiens, Cx. territans 0.87 An. maculipennis s.l., An. superpictus, Cx. theileri, Cx. tritaeniorhynchus 0.87 An. maculipennis s.l., Cx. territans, Cx. tritaeniorhynchus 0.87 Cx. pipiens 0.87 An. maculipennis s.l., Cx. mimeticus, Cx. pipiens 0.58 Ae. vexans 0.29 Cx. hortensis, Cx. pipiens, Cx. tritaeniorhynchus 0.29 Cx. mimeticus, Cx. pipiens, Cx. territans, Cx. tritaeniorhynchus 0.29 An. superpictus, Cx. hortensis, Cx. pipiens, Cx. tritaeniorhynchus 0.29 Ae. vexans,, Cx. mimeticus, Cx. pipiens, Cs. annulata, Cs. morsitans 0.29 An. superpictus, Cx. mimeticus, Ur. unguiculata 0.29 Total 100 Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 44 Table 4. Continued…. Discussion Larval habitat characteristics and ecology In the present investigation, five species of the genus Anopheles were found and iden- tified based on the morphological characters of larvae in different aquatic habitats in Gui- lan Province. While the tree-hole An. plum- An. maculipennis s.l. An.’hyrcanus’, Cx. tritaeniorhynchus 27.82 An.’hyrcanus’, Cx. mimeticus 14.74 Cx. pipiens 10.46 An.’hyrcanus’, Cx. mimeticus, Cx. tritaeniorhynchus 10.23 An.’hyrcanus’, Cx. theileri, Cx. tritaeniorhynchus 9.16 An.’hyrcanus’, An. superpictus, Cx. mimeticus, Cx. tritaeniorhynchus 3.69 An.’hyrcanus’ 3.33 Cx. mimeticus 2.62 Alone 2.02 An.’hyrcanus’, Cx. hortensis, Cx. tritaeniorhynchus 1.90 An. claviger, An.’hyrcanus’ 1.78 Cx. theileri 1.78 Cx. theileri, Cx. tritaeniorhynchus 1.43 An.’hyrcanus’, Cx. pipiens, Cx. theileri 1.43 An.’hyrcanus’, Cx. territans, Cx. tritaeniorhynchus 1.19 An. claviger, Cx. territans, Cx. tritaeniorhynchus 1.07 An.’hyrcanus’, Cx. mimeticus, Cx. pipiens 0.71 An.’hyrcanus’, Cx. pipiens, Cx. territans 0.71 Cx. pipiens, Cx. tritaeniorhynchus 0.71 An.’hyrcanus’, Cx. pipiens, Cx. theileri, Cx. tritaeniorhynchus, Cs. longiareolata 0.59 An.’hyrcanus’, An. superpictus, Cx. theileri, Cx. tritaeniorhynchus 0.59 An.’hyrcanus’, Cx. territans 0.48 An. claviger, An.’hyrcanus’, Ae. vexans, Cx. pipiens 0.36 An. superpictus, Cx. tritaeniorhynchus 0.36 Cx. pipiens, Cx. territans 0.36 An.’hyrcanus’, Cx. pipiens, Cx. theileri, Cx. tritaeniorhynchus 0.12 Cx. hortensis 0.12 An.’hyrcanus’, Cx. pipiens, Cx. tritaeniorhynchus 0.12 Cs. morsitans 0.12 Total 100 An. plumbeus Alone 62.87 Oc. echinus, Oc. geniculatus 18.81 Oc. echinus 16.34 Cx. pipiens 1.98 Total 100 An. superpictus An.’hyrcanus’, Cx. hortensis, Cx. pipiens, Cx. tritaeniorhynchus 26.23 An. claviger, Cx. hortensis, Cs. longiareolata 19.67 An. maculipennis s.l., Cx. tritaeniorhynchus 19.67 Cx. theileri, Cx. tritaeniorhynchus 11.48 An.’hyrcanus’, An. maculipennis s.l., Cx. mimeticus, Cx. tritaeniorhynchus 11.48 An.’hyrcanus’, Cx. mimeticus, Ur. unguiculata 9.84 An.’hyrcanus’, An. maculipennis s.l., Cx. theileri, Cx. tritaeniorhynchus 1.63 Total 100 Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 45 beus has its own special habitat and way of adaptation, other species have shared some common larval habitat characteristics such as temporary, stagnant, and clear water. However every species shows some preferred characteristics in comparison with others as follow: An. superpictus prefers the habitats without vegetation (65.6%); An. claviger and An. superpictus more adapt the habitats with mud (73.7% and 67.8%, respectively) or gravel (17.9% and 32.2%) substrate, respec- tively, whereas An. ’hyrcanus’ and An. ma- culipennis s.l. are less conservative in this regard; An. claviger lays the eggs often (66.0%) in shaded area; An. claviger and An. superpictus choose mostly natural habitats (97.9% and 100%), especially rain pools (66.3% and 67.2%), however An. ’hyrcanus’ and An. maculipennis s.l. show more diverse habitats and were also collected from artificial habitats (16.8% and 16.4%), espe- cially rice fields (79.4% and 61.6%) (Table 3). These explain why An. maculipennis s.l. and An.’hyrcanus’ are the most abundant species in the studied area where rice fields and irrigation channels are numerous and vast (Table 1). Macan (1950) found An. claviger in "spring pools, sometimes exposed to the sun, and in slow-flowing reaches and isolated pools of the upper parts of streams where these were heavily shaded" with water temperature 14– 16° C (except for one occasion 20° C) in Iraq and western Iran. Dow (1953) found An. claviger in "a small, shallow and shaded stream, with much emergent and partly submerged vegetation including some grass" with Cx. pipiens in Maragheh of northwestern Iran. Horsfall (1955) mentioned that An. claviger is an associate of An. cinereus Theobald (as An. hispaniola), An. maculipennis, An. marteri Senevet and Prunnelle, An. superpictus, Cs. longiarolata, Cs. morsitans, Cx. laticinctus Edwards, Cx. mimeticus, Cx. pipiens, Cx. territans (as Cx. apicalis), and Cx. tritaenio- rhynchus and noted that the larvae have been collected from water with temperature 12– 15° C and even beneath the ice during winter and larval sites include: small ponds, flow- ing stream margins, springs, cisterns, foun- tains, coastal areas and marshes, shaded rain barrels, roadside ditches, even sewages, and especially wells in different areas of its distribution. Zaim (1987a) collected An. claviger from stream edge in Kashan. In the present study, An. marteri and Cx. laticinc- tus were not found in Guilan Province (Azari-Hamidian et al. 2002b, 2004a, 2005, Azari-Hamidian 2007b) and An. cinereus has not been collected in Iran (Azari-Hamidian 2007a). Culiseta morsitans was found in the province, not in association with An. claviger, however other mentioned species were col- lected with this species (Tables 2 and 4). As Horsfall (1955) expressed wells are the important habitats of An. claviger larvae, in this study among man-made habitats it was found in wells only, although the species was collected mostly from natural habitats (97.9%) (Table 3). Unfortunately, there is little information about the larval habitat characteristics and ecology of An. hyrcanus and An. pseudopic- tus in details because of their problematic dif- ferentiation in larval stage (see Taxonomic note). Dow (1953) collected only An. pseudopictus (as An. hyrcanus var. pseudopictus) in north- ern Iran based on rearing immature stages from "fairly deep channels in river bed with emergent vegetation and surface debris", "rice fields", "quiet river channel", "canal with emer- gent grass along banks", "small brook below spring with mats of glove-like alga" with An. maculipennis, An. melanoon (as An. subal- pinus) and Cx. tritaeniorhynchus. Horsfall (1955) stressed An. algeriensis, An. coustani Laveran, An. sacharovi, An. sergentii (Theo- bald) and rarely An. claviger, An. superpic- tus, Cs. annulata, Cx. perexiguus Theobald, Oc. caspius s.l., and Oc. detritus as the as- sociated species of An. ‘hyrcanus’ in Palestine. Azari-Hamidian et al. (2002a) found An. ‘hyr- Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 46 canus’ larvae with those of An. maculipennis s.l., Ae. vexans, Cx. mimeticus, Cx. pipiens, Cx. theileri, and Cx. tritaeniorhynchus often in rice fields and also ground pools in Rasht County. Among mentioned species, Anopheles coustani is not recorded in Iran (Azari- Hamidian 2007a) and An. algeriensis, An. sacharovi, An. sergentii, Cx. perexiguus, and Oc. detritus were not found in Guilan Province in this study (Azari-Hamidian et al. 2002b, 2002c, 2004a, 2005, Azari-Hamidian 2007b) and Oc. caspius s.l. was collected only as adult (Azari-Hamidian et al. 2002c). Other species larvae were found with An. ‘hyrcanus’ larvae. In the present study, An. hyrcanus, with dark hindtarsomere 4, except at tip, and completely dark hindtarsomere 5, reared from pupae was an associate of An. maculipennis s.l., An. pseudopictus, Ae. vex- ans, Cx. mimeticus, Cx. pipiens, and Cx. tri- taeniorhynchus, which were reared from pu- pae or collected as larvae from the same lar- val habitats in ground pools with emergent vegetation and mud or sand substrate and river bed pools with submerged vegetation and gravel substrate both habitats were ex- posed to sunlight. Also, reared An. pseu- dopictus, with completely pale hindtarsomere 4 and completely dark hindtarsomere 5, was with An. claviger, An. maculipennis s.l., An. hyrcanus, Ae. vexans, Cx. mimeticus, Cx. pipiens, Cx. territans, Cx. theileri, Cx. tri- taeniorhynchus in river edge without vegeta- tion, with mud substrate in shaded area, river bed pool with submerged vegetation and gravel substrate in full sunlight situation, rice irri- gation channel without vegetation with sand substrate in shaded area, and ground pool with emergent vegetation and mud substrate in full sunlight situation. Both species were collected from habitats with clear, transient, and stagnant water. In general, two species occur sympatrically in northern Iran; however An. pseudopictus is much more abundant than An. hyrcanus (Dow 1953, Azari-Hamidian et al. 2001, 2006) and the main larval habitats are: river bed pools, rain pools, and rice fields (Table 3). Ghanbari et al. (2005) men- tioned some physical and chemical factors of the larval habitats of Anopheles including An. hyrcanus in Iranshahr, southeastern Iran, however most probably their identification is not correct and needs to be verified. Because, the Oriental species of the Hyrcanus Group, An. peditaeniatus, has been found in south- eastern Iran and old records of the Oriental An. nigerrimus Giles in southern and south- eastern Iran using old keys and the record of the Palaearctic An. hyrcanus in southeastern Iran might be misidentifications and need to be verified (Azari-Hamidian et al. 2006). There is little data about the larval breeding places and ecology of the Maculipennis Group species in details in Iran, because the species were not identified carefully in larval stage or using egg pattern in the most of ecological investigations and most of recorded species based on PCR technique were collected as adult (see Taxonomic note). Dow (1953) iden- tified An. maculipennis based on egg col- lected in "rice field", "small spring in ditch", "shady irrigation reservoir", "shady irrigation ditch", "wide irrigation ditch with floating plants, debris and algae, also emergent grass and sedge", "puddle in small irrigation ditch beside road", and "small stream with much floating and emergent vegetation" in north- western Iran and the Caspian Sea littoral (Ma- zandaran Province) and the associated spe- cies were An. ‘hyrcanus’, An. sacharovi, An. superpictus, Cx. hortensis, and Cx. theileri. Also he identified An. melanoon (as An. melanoon subspecies subalpinus) collected in "quiet river channel", "canal with emergent grass along bank", "small brook below spring with mats of glove-like alga", "rain barrel", and "shallow pond" in Guilan Province and the associated species were An. ‘hyrcanus’ and Cx. tritaeniorhynchus. Anopheles sa- charovi was collected in a "shady irrigation reservoir" and in a "pasture" in northwestern Iran. Horsfall (1955) mentioned An. macu- Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 47 lipennis s.l. an associate of An. ‘hyrcanus’, An. sergentii, An. superpictus, Cs. annulata, Cx. laticinctus, Cx. pipiens, and Ur. un- guiculata and added rarely two or more spe- cies of the group were found in the same habitat at the same time such as An. atropar- vus with An. messeae and An. maculipennis with An. melanoon (as An. subalpinus). Mousa- kazemi et al. (2000) collected An. maculipen- nis s.l. larvae with those of An. superpictus, Ae. vexans, Cs. longiareolata, Cx. pipiens, Cx. perexiguus, Cx. theileri, Oc. caspius s.l. from rice fields in Isfahan Province. Yag- hoobi-Ershadi et al. (2001) did not identified An. sacharovi based on egg pattern, larval chaetotaxy, or PCR in Parsabad of northern Ardebil Province; however they studied its larvae based on the high abundance of the adults in the studied area. They noted very high salinity of the larval habitat waters (263–414 mg/l) and An. superpictus as associated spe- cies. Anopheles mculipennis have been found in northern Ardebil Province (Parsabad and Bil-e-Savar Counties), however with very low density (Yaghoobi-Ershadi et al. 2001, Azari-Hamidian et al. 2009). Azari-Hamidian et al. (2002a) collected An. maculipennis s.l. larvae in rice fields, irrigation channels, ground pools, in sunlight or shaded situation, with or without vegetation, with mud substrate and clear water in Rasht County of Guilan Province. The associated larvae were An. ‘hyr- canus’, Ae. vexans, Cx. pipiens, Cx. theileri, and Cx. tritaeniorhynchus. In the present investigation, among the mentioned species; An. sacharovi, An. sergentii, Cx. laticinctus, and Cx. perexiguus were not found in the province (Azari-Hamidian et al. 2002b, 2004a, 2005, Azari-Hamidian 2007b) and Cs. annu- lata and Ur. unguiculata larvae were not found with An. maculipennis s.l. (Azari- Hamidian et al. 2004b, Azari-Hamidian 2005), however other species were in association with this species (Tables 2 and 4). In general, An. maculipennis is the most abundant spe- cies of the Maculipennis Group in Guilan Prov- ince (Zolotarev 1945, Dow 1953, de Zulueta et al. 1957, Azari-Hamidian et al. 2004a) and its main larval habitats, such as An. ‘hyr- canus’, are: river bed pools, rain pools, and rice fields (Table 3). The tree-hole mosquito An. plumbeus also has been found in different habitats other than those in soil including: wells, cisterns, vases and other domestic containers, even dung pits, and flowerpots. The most favor- able temperature is 15-18° C; however the species has tolerated to freeze and -6° C to -8° C in the laboratory. Associated species are Ae. aegypti (Linnaeus), Cs. annulata, Cx. pipiens, Ochlerotatus geniculatus, Oc. ech- inus, and Orthopodomyia pulcripalpis (Ron- dani) (Horsfall 1955). There is no record of Or. pulcripalpis in Iran and Ae. aegypti has not been found in the country for more than fifty years (Azari-Hamidian 2007a). In the present investigation, Cs. annulata was col- lected in Guilan Province (Azari-Hamidian et al. 2003b, Azari-Hamidian 2005), how- ever not in association with An. plumbeus. Anopheles plumbeus was associated with other three species (Tables 2 and 4). Most of An. plumbeus larvae were collected from the tree holes and only in one occasion four lar- vae were collected with Cx. pipiens in a dis- carded tire (Table 3). The larvae were only collected when rain season started, October and November, and tree-holes included wa- ter and the weather was cooler (Azari- Hamidian 2003, 2006). That is why the mean temperature of habitat water showed significant difference from other species. Macan (1950) stressed "the edges of stony streams" as "the classic habitat" of An. superpictus, however he added larvae are found in "any available breeding places where the water is clean, exposed to sunlight and moderately shallow" in western Iran. Dow (1953) found An. superpictus larvae with An. maculipennis, and probably An. sa- charovi, Cx. hortensis, Cx. theileri, and Ur. unguiculata in different kinds of habitats in- Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 48 cluding; rivers, streams, pools, pastures, springs, irrigation ditches in different areas of Iran. Horsfall (1955) said the associated species of the species are; An. cinereus (as An. hispaniola), An. claviger, An. sacharovi, An. sergentii, Cx. laticintus, Cx. mimeticus, and Ur. unguiculata and mentioned that three main larval breeding places of An. su- perpictus are: "clear shallow water over rocky bottom", "rivers of clear water over sandy bottom", and "hill streams of shallow water over mud bottom". The larvae passed the winter in pit wells where the temperature was 5–15°C in Greece. Yaghoobi-Ershadi et al. (1986) found An. superpictus larvae in habitats such as river edges (80%), ground pools (15%), and pastures (5%) which were often transient (62.5%) and stagnant (100%), without (50%) or with submerged (22.8%) vegetation, in partial (56.3%) or full (37.4%) sunlight, with mud (56.2%) and sand (43.8%) substrate. Also they noted associated larvae: An. dthali, An. fluviatilis s.l., An. multicolor, An. stephensi, and An. turkhudi in Minab. Mousakazemi et al. (2000) collected An. superpictus larvae with those of An. macu- lipennis s.l., Ae. vexans, Cs. longiareolata, Cx. pipiens, Cx. perexiguus, Cx. theileri, Oc. caspius s.l. from rice fields in Isfahan Province. Yaghoobi-Ershadi et al. (2001) found An. superpictus larvae with An. sacharovi in Ardebil Province. Ghanbari et al. (2005) studied some physical and chemical factors of the larval habitats of Anopheles larvae in Iranshahr including An. superpictus. Among mentioned species An. cinereus has not been found in Iran (Azari-Hamidian 2007a) and An. dthali, An. fluviatilis s.l., An. multicolor, An. sergentii, An. stephensi, An. turkhudi, Cx. laticinctus, and Cx. perexiguus in Guilan Province (Azari-Hamidian et al. 2002b, 2004a, 2005, Azari-Hamidian 2007b). In the present study, Anopheles sacharovi was not found in the province (Azari-Hamidian et al. 2002b, 2004a) and Ae. vexans larvae were not in association with An. superpictus larvae (Azari- Hamidian 2006) and Oc. caspius s.l. was found only as adult (Azari-Hamidian et al. 2002c). Other mentioned species were as- sociated with An. superpictus larvae in the province (Tables 2 and 4). In parallel to the results of many researchers (Macan 1950, Dow 1953, Horsfall 1955, Yaghoobi-Ershadi et al. 1986, Zaim 1987a), in this study An. superpictus was found in natural habitats (100%) and mostly exposed to sun (88.5%), with clear (100%) water, without vegetation (65.6%), and mud (67.8%) substrate (Table 3). Finding in rice fields (artificial habitat) by Mousakazemi et al. (2000) in Isfahan Province seems to be to some extent unusual and interesting. Larval habitat water pH, electric conduc- tivity (EC), turbidity, dissolved oxygen, and different organic and inorganic compounds, other aquatic associated animals especially predators, and the life tables of different mosquito species are items, which should be considered in future studies. Checklist of the mosquitoes of Guilan Prov- ince Azari-Hamidian et al. (2002b) found 23 species of mosquitoes representing seven genera based on morphological characters including egg pattern (for the Maculipennis Group) in Guilan Province. In this list three species An. superpictus, Oc. echinus, and Ur. unguiculata were recorded in the prov- ince and Cs. morsitans and its subgenus (Culicella) in Iran for the first time. Five species including An. atroparvus and An. sacharovi of the Maculipennis Group, An. algeriensis, Cs. subochrea, and Cx. torren- tium, which were recorded in Guilan Prov- ince before (Danilov 1975, Saebi 1987, Zaim 1987b, Dinparast-Jadid et al. 2001), were not found by Azari-Hamidian et al. (2002b). After that, Sedaghat et al. (2003) and Gholi- zadeh et al. (2004) reported An. persiensis and An. labranchiae of the Maculipennis Group based on PCR technique in the prov- Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 49 ince, respectively. In general, all seven spe- cies of the Maculipennis Group recorded in Iran (Azari-Hamidian 2007a) have been found in Guilan Province. Anopheles at- roparvus, An. labranchiae, An. maculipen- nis, An. messeae, An. persiensis, An. sa- charovi are presented in the province and Iran based on DNA data and their sequences are available in GenBank (Sedaghat et al. 2003, Gholizadeh et al. 2004, Gholizadeh et al. 2005, Djadid et al. 2007), however An. melanoon (also as An. subalpinus) has been found only using egg pattern (Zolotarev 1945, Dow 1953, de Zulueta et al. 1957, Azari-Hamidian et al. 2002b). Dow (1953) and Djadid et al. (2007) found An. sacharovi in Guilan Province based on egg pattern (in Hassan Kiadeh) and DNA sequence, respec- tively, however it seems, as Dow (1953) noted, the species is quite rare in the prov- ince. Azari-Hamidian et al. (2002b, 2004a) identified three species An. maculipennis, An. melanoon, and An. messeae based on egg pattern in Guilan Province. During his studies, Saebi (1987) did not find An. al- geriensis in Iran and mentioned it a very rare species which had been recorded in Lahijan and Fooman (Fuman) of Guilan Province before. This species was not found by Azari- Hamidian et al. (2002b, 2004b) either; how- ever its occurrence in the province seems to be possible, because of the presence of its preferred habitats; reedy swamps (Horsfall 1955). Based on the second Internal Tran- scribed Spacer (ITS2) and Cytochrome c Oxidase subunit I (COI) sequence data, re- cently Oshaghi et al. (2007, 2008) reported three genotypes named X, Y, and Z within An. superpictus in Iran. Genotype X was found in all parts of the country except for the southeastern areas. It seems that the An. superpictus specimens of Guilan Province belong to genotype X, however it needs to be confirmed by more investigation. Zaim (1987b) reported Cs. subochrea in Guilan Province; however Azari-Hamidian et al. (2002b, 2003b) found only Cs. annulata. These two close and rare species are very similar in larval stage; however they are easily distinguishable as adult. It seems the occurrence of Cs. subochrea in Guilan Province is possible, especially in view of the recent finding of both species in Ardebil Province, northwestern Iran (Azari-Hamidian et al. 2009). There is little information about taxonomy and distribution of three close species Cs. alaskaensis (Ludlow), Cs. annu- lata, and Cs. subochrea in Iran (Azari- Hamidian et al. 2003b, Azari-Hamidian 2005). Recently, Cs. subochrea and Cs. longiareo- lata were found in Sanandaj County of Kur- distan Province (Moosa Kazemi et al. 2010). Danilov (1975) was the unique reliable re- cord of Cx. torrentium in Rasht of Guilan Province and Iran. Recently the larvae of this species was found in Ardebil Province (Azari-Hamidian et al. 2009) and the moun- tainous areas of the western part of Guilan Province (Heyran) (with Cx. pipiens; Azari- Hamidian, unpublished data), thus its occur- rence in Iran, Guilan Province, and for the first time in Ardebil Province was verified. All 30 species, which have been discussed above, were mentioned in the checklist in the present article. There is a unique record of Cx. quinquefasciatus Say in Enzeli of Guilan Province by Harbach (1988). There is no more information about this record (Har- bach, personal communication). This is out of the usual distribution of this species that occurs in southern Iran (Zaim 1987b), so it is not mentioned in the checklist. Taxonomy, distribution, and ecology of the Pipiens Subgroup of the Pipiens Group of Culex (Culex) need to be studied extensively in Iran. Beklemishev and Shipitzina (1947) found An. marteri in the valley of the river Yuzbashchai between Qazvin (Qazvin Prov- ince) and Rasht (Guilan Province); however there is no record of this species in Guilan Province. The species complexes and the mosquito fauna of the forest areas of the Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 50 province are important subjects to study in future taxonomic investigations. Acknowledgements The author is grateful to J Minar, Institute of Parasitology, Czech Republic, for study- ing specimens, JF Reinert, Center for Medi- cal, Agricultural and Veterinary Entomol- ogy, Gainesville, for providing some articles, J Jezek, Natural History Museum of Prague, Prague, I Gelbic, Institute of Entomology, Czech Republic, F Rettich, National Institute of Public Health, Prague, for their important correspondences and helps. The author also thanks MA Oshaghi, Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, for reviewing the manuscript and S Omidi, School of Health, Guilan University of Medical Siences, Rasht, Iran, for assisting in statistical analy- sis. This study received financial support from Guilan University of Medical Sciences. References Azari-Hamidian S, Yaghoobi-Ershadi MR, Javadian E (2001) A faunistic study of mosquito (Diptera: Culicidae) fauna in Rasht County (Iran). Modarres J Med Sci. 3: 65–70 [Persian with English ab- stract]. Azari-Hamidian S, Yaghoobi-Ershadi MR, Javadian E (2002a) The distribution and larval habitat characteristics of mosquitoes (Diptera: Culicidae) in Rasht County (Guilan Province, Iran). Modarres J Med Sci. 4: 87–96 [Persian with English abstract]. Azari-Hamidian S, Joeafshani MA, Mosslem M, Rassaei AR (2002b) Taxonomic survey of mosquitoes (Diptera: Culici- dae) in Guilan Province with reporting of a subgenus new to Iranian mosquito fauna. The 15th Iranian Plant Protection Congress, 7–11 September 2002, Razi University of Kermanshah, Kerman- shah, Iran. pp. 319–320 [Persian with English abstract]. Azari-Hamidian S, Joeafshani MA, Rassaei AR, Mosslem M (2002c) Mosquitoes of the genus Aedes (Diptera: Culicidae) in Guilan. J Med Fac Guilan Univ Med Sci. 11(43): 29–39 [Persian with Eng- lish abstract]. Azari-Hamidian S (2003) On the tree hole mosquito (Diptera: Culicidae) fauna, ecology and medical importance in Guilan Province. The 4th National Ira- nian Congress of Parasitology and Parasitic Diseases, 13–16 October 2003, Mashhad, Iran. p. 293 [Persian with English abstract]. Azari-Hamidian S, Joeafshani MA, Mosslem M, Rassaei AR (2003a) Adult mos- quito habitats and resting-places in Gui- lan Province (Diptera: Culicidae). Ha- kim. 6: 55–62 [Persian with English abstract]. Azari-Hamidian S, Joeafshani MA, Rassaei AR, Mosslem M (2003b) Faunistic studies on the genus Culiseta (Diptera: Culicidae) in Guilan Province. J Ker- man Univ Med Sci. 10: 225–233 [Per- sian with English abstract]. Azari-Hamidian S, Joeafshani MA, Rassaei AR, Mosslem M, Mousavi-Eivanaki E (2004a) Mosquito fauna of the genus Anopheles (Diptera: Culicidae) in Gui- lan Province. Modarres J Med Sci. 6: 11–22 [Persian with English abstract]. Azari-Hamidian S, Joeafshani MA, Mosslem M, Rassaei AR (2004b) Notes on Co- quillettidia richiardii and Uranotaenia unguiculata (Diptera: Culicidae) in Guilan Province. J Med Fac Guilan Univ Med Sci. 13(51): 1–9 [Persian with English abstract]. Azari-Hamidian S (2005) Larval habitat characteristics of mosquitoes of the ge- nus Culiseta Felt, 1904 (Diptera: Culi- Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 51 cidae) in the Caspian Sea littoral, Iran. Zool Middle East. 36: 59–66. Azari-Hamidian S, Joeafshani MA, Mosslem M, Rassaei AR (2005) Mosquito fauna and distribution of the genus Culex (Diptera: Culicidae) in Guilan Prov- ince. Pajouhesh and Sazandegi. 68: 39– 45 [Persian with English abstract]. Azari-Hamidian S (2006) On the ecology of the tribe Aedini (Diptera: Culicidae) larvae in the Caspian sea littoral, Iran. The 11th International Congress of Parasitology, 6–11 August 2006, Glas- gow, Scotland, UK. Azari-Hamidian S, Abai MR, Ladonni H, Vatandoost H, Akbarzadeh K (2006) Anopheles peditaeniatus (Leicester) new to the Iranian mosquito fauna with notes on Anopheles hyrcanus group in Iran. J Am Mosq Control Assoc. 22: 144–146. Azari-Hamidian S (2007a) Checklist of Ira- nian mosquitoes (Diptera: Culicidae). J Vector Ecol. 32: 235–242. Azari-Hamidian S (2007b) Larval habitat characteristics of mosquitoes of the ge- nus Culex (Diptera; Culicidae) in Gui- lan Province, Iran. Iran J Arthropod- Borne Dis. 1: 9–20. Azari-Hamidian S, Yaghoobi-Ershadi MR, Javadian E, Mobedi I, Abai MR (2007) Review of dirofilariasis in Iran. J Med Fac Guilan Univ Med Sci. 15(60): 102–113 [Persian with English abstract]. Azari-Hamidian S, Yaghoobi-Ershadi MR, Javadian E, Abai MR, Mobedi I, Lin- ton Y-M, Harbach RE (2009) Distri- bution and ecology of mosquitoes in a focus of dirofilariasis in northwestern Iran, with the first finding of filarial larvae in naturally infected local mos- quitoes. Med Vet Entomol. 23: 111–121. Beklemishev V, Shipitzina N (1947) Ano- pheles marteri in the north-western Iran. Med Parasitol Parasit Dis. 16: 66– 67 [Russian with English abstract]. Danilov VN (1975) Mosquito fauna of northwestern Iran. Med Parasitol Para- sit Dis. 44: 732 [Russian with English abstract]. Darsie RF, Samanidou-Voyadjoglou A (1997) Keys for the identification of the mosquitoes of Greece. J Am Mosq Control Assoc. 13: 247–254. de Zulueta J, Jolivet P, Thymakis K, Caprari P (1957) Seasonal variations in sus- ceptibility to DDT of Anopheles macu- lipennis in Iran. Bull WHO.16: 475–479. Dinparast-Jadid N, Kordiev M, Townson H (2001) Molecular key to Anopheles maculipennis species complex and its application for malaria control in the costal area of Caspian Sea. The 3rd Ira- nian Congress of Medical Parasitology, Sari, Iran. p. 108 [Persian]. Djadid ND, Gholizadeh S, Tafsiri E, Romi R, Gordeev M, Zakeri S (2007) Mo- lecular identification of Palearctic members of Anopheles maculipennis in northern Iran. Malaria J. 6: 6–15. Djadid ND, Jazayeri H, Gholizadeh S, Rad ShP, Zakeri S (2009) First record of a new member of Anopheles Hyrcanus Group from Iran: molecular identifica- tion, diagnosis, phylogeny, status of kdr resistance and Plasmodium infec- tion. J Med Entomol. 46: 1084–1093. 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: 21–27. Dow RP (1953) Notes on Iranian mosqui- toes. Am J Trop Med Hyg. 2: 683–695. Edrissian GH (2006) Malaria in Iran: Past and present situation. Iran J Parasitol. 1: 1–14. Eshghi N, Motabar M, Javadian E, Mano- uchehri AV (1976) Biological features of Anopheles fluviatilis and its role in Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 52 the transmission of malaria in Iran. Trop Geogr Med. 28: 41–44. Eshghy N (1977) Anopheles multicolor Cambouliu and its role in the transmis- sion of malaria in Iran. J Entomol Soc Iran. 4: 87–88 [Persian with English abstract]. Eslami A (1997) Veterinary Helminthology. Vol. 3. Nematoda and Acanthocephala. Tehran University Publications, Tehran [Persian]. Ghanbari MR, Rakhsh Khorshid A, Salehi M, Hassanzehi A (2005) The study of physical and chemical factors affecting breeding places of Anopheles in Iran- shahr. Tabib-e-Shargh, J Zahedan Univ Med Sci. 7: 221–227 [Persian with Eng- lish abstract]. Gholizadeh S, Tafsiri E, Zakeri S, Dinparast- Jadid N (2004) Molecular identifica- tion of a new species to Iranian ano- pheline fauna of Anopheles maculi- pennis complex (Diptera: Culicidae). The 13th Iranian Congress on Infec- tious Diseases and Tropical Medicine, 11–15 December 2004, Tehran, Iran. p. 197 [Persian]. Gholizadeh S, Tafsiri E, Zakeri S, Dinparast Djadid N (2005) Anopheles maculi- pennis complex (Diptera: Culicidae) in Guilan Province of Iran. The 13th Ira- nian Biology Conference and the First International Conference of Biology, 23–25 August 2005, Rasht, Iran. p. 92. Harbach RE (1988) The mosquitoes of the subgenus Culex in southwestern Asia and Egypt (Diptera: Culicidae). Con- trib Am Ent Inst. 24: 1–240. Harbach RE (2007 onwards) Mosquito Taxonomic Inventory. Available at: http://mosquito-taxonomic-inventory. info/ (accessed 25 May 2011). Horsfall WR (1955) Mosquitoes. Their Bio- nomics and Relation to Disease. Haf- ner Publishing, New York. Lotfi MD (1973) Iranian species of genus Culex II Report of four species of lar- vae and 14 adult species. Bull Soc Pathol Exot. 66: 204–207. Macan TT (1950) Anopheles and Malaria in the Near East III. The Anopheline Mosquitoes of Iraq and North Persia. London School of Hygiene and Tropi- cal Medicine Research Memoir No 7, HK and Lewis Co Ltd, London. Manouchehri AV, Rohani F (1975) Notes on the ecology of Anopheles dthali Patton in southern Iran. Ann Trop Med Para- sit. 69: 393–397. Manouchehri AV, Javadian E, Eshghy N, Motabar M (1976) Ecology of Anopheles stephensi Liston in southern Iran. Trop Geogr Med. 28: 228–232. Marsh F (1933) New species of Anopheles (Myzomyia group) from south-west Persia. Stylops. 2: 193–197. Momeni S, Manouchehri AV, Zaim M, Mottaghi M (1992) The distinguish between the species of Anopheles macu- lipennis complex by using of egg char- acters in Gilan and Mazandaran Prov- inces, northern areas of Iran 1990–1991. Iranian Congress of Malaria, 22–26 February 1992, Zahedan, Iran. p. 29. Moosa Kazemi SH, Karimian F, Davari B (2010) Culicinae mosquitoes in Sanan- daj County, Kurdistan Province, west- ern Iran. J Vector Borne Dis. 47: 103-107. Mousakazemi S, Zaim M, Zahraii A (2000) Fauna and ecology of Culicidae of the Zarrin-Shahr and Mobarakeh area in Isfahan Province. Armaghan Danesh, J Yasuj Univ Med Sci. 5: 46–54 [Persian with English abstract]. Naficy K, Saidi S (1970) Serological survey on viral antibodies in Iran. Trop Geogr Med. 2: 183–188. Oshaghi MA, Shemshad Kh, Yaghobi-Er- shadi MR, Pedram M, Vatandoost H, Abaie MR, Akbarzadeh K, Mohtarami F (2007) Genetic structure of the ma- Iran J Arthropod-Borne Dis, 2011, 5(1): 37–53 S Azari-Hamidian: Larval Habitat Characteristics … 53 laria vector Anopheles superpictus in Iran using mitochondrial cytochrome oxidase (COI and COII) and morphol- ogic markers: A new species complex? Acta Trop. 101: 241–248. Oshaghi MA, Yaghobi-Ershadi MR, Shem- shad Kh, Pedram M, Amani H (2008) The Anopheles superpictus complex: introduction of a new malaria vector complex in Iran. Bull Soc Pathol Exot. 101: 429–434. Reinert JF (2001) Revised list of abbrevia- tion for genera and subgenera of Culi- cidae (Diptera) and the notes on ge- neric and subgeneric changes. J Am Mosq Control Assoc. 17: 51–55. Saebi ME (1987) Morphological Study on Anopheline Larvae and their Distribu- tion in Iran. [PhD Dissertation]. School of Public Health, Tehran University of Medical Sciences, Tehran, Iran [Per- sian]. Saidi S, Tesh R, Javadian E, Nadim A (1976) The prevalence of human in- fection of West Nile in Iran. Iran J Publ Health. 5: 8–14. Sedaghat MM, Linton Y-M, Oshaghi MA, Vatandoost H, Harbach RE (2003) The Anopheles maculipennis complex (Dip- tera; Culicidae) in Iran: molecular char- acterization and recognition of a new species. Bull Entomol Res. 93: 527– 535. Service MW (1993) Mosquitoes (Culicidae). In: Lane RP, Crosskey RW (Eds) Medical Insects and Arachnids. Chap- man and Hall, London, pp. 120–240. Shahgudian ER (1960) A key to anophelines of Iran. Acta Med Iran. 3: 38–48. WHO (1975) Manual on Practical Entomol- ogy in Malaria. Part II. Methods and Techniques. WHO, Geneva. Yaghoobi-Ershadi MR, Zaim M, Manouche- hri AV (1986) Studies on the biology of the mosquitoes in the district of Mi- nab, Hormozgan Province, Iran, (1983–84)1-Characteristcs of the larval breeding sites. J Environ Stud. 13: 17– 39 [Persian with English abstract]. Yaghoobi-Ershadi MR, Namazi J, Piazak N (2001) Bionomics of Anopheles sa- charovi in Ardebil Province, north- western Iran during a larval control program. Acta Trop. 78: 207–215. Zaim M, Cranston PS (1986) Checklist and keys to the Culicinae of Iran (Diptera; Culicidae). Mosq Syst. 18: 233–245. Zaim M (1987a) The mosquito fauna of Ka- shan, public health importance and con- trol. Desert, Sci Res. 18: 1-41 [Persian]. Zaim M (1987b) The distribution and larval habitat characteristics of Iranian Cu- licinae. J Am Mosq Control Assoc. 3: 568–573. Zaim M, Subbarao SK, Manouchehri AV, Cochrane AH (1993) Role of Anophe- les culicifacies s.l. and An. pulcherri- mus in malaria transmission in Ghass- reghand (Baluchistan), Iran. J Am Mosq Control Assoc. 9: 23–26. Zaini A, Janbakhsh B, Manouchehri A (1975) Characteristics of breeding places of Anopheles stephensi in city of Bandar-Abbas. Iran J Publ Health. 4: 114–119 [Persian with English ab- stract]. Zolotarev ER (1945) Anopheles maculipen- nis of northern Iran. Med Parasitol Parasit Dis. 14: 50–57 [Russian].