NORTH-WESTERN JOURNAL OF ZOOLOGY 11 (1): 76-85 ©NwjZ, Oradea, Romania, 2015 Article No.: 141208 http://biozoojournals.ro/nwjz/index.html
The fauna, habitats, and affinity indices of mosquito larvae (Diptera: Culicidae) in Central Iran
Hassan LADONNI1, Shahyad AZARI-HAMIDIAN2,*, Mostafa ALIZADEH3, Mohammad-Reza ABAI1 and Hassan BAKHSHI1
1. Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. 2. School of Health, Guilan University of Medical Sciences, Rasht, Iran. 3. Department of Diseases Control, Deputy of Health, Isfahan University of Medical Sciences, Isfahan, Iran. * Corresponding author, S. Azari-Hamidian, Tel: +98 131 3229599, Fax: +98 131 3234155, E-mail: [email protected]
Received: 04. August 2013 / Accepted: 23. December 2013 / Available online: 01. January 2015 / Printed: June 2015
Abstract. Malaria and West Nile virus infections are rampant to Isfahan Province, Central Iran. These mosquito-borne diseases along with the nuisance caused by female adult bitings are troublesome throughout mosquito active season. During July–October 2003, May–September 2004 and 2006, we collected mosquito larvae from Isfahan using dipping method, to study their fauna, habitat, and affinity indices. Collected larvae were preserved in lactophenol and mounted using de Faure’s medium prior to microscopy study. Larvae were identified based on morphological characters. Out of 1659 third- and fourth-instar larvae, we discovered 15 species representing five genera including; Anopheles dthali, An. maculipennis s.l., An. marteri, An. multicolor, An. superpictus, An. turkhudi, Aedes vexans, Culex hortensis, Cx. mimeticus, Cx. perexiguus, Cx. pipiens, Cx. territans, Cx. theileri, Culiseta longiareolata, and Ochlerotatus caspius s.l. This is the first record of An. dthali, An. marteri, and An. turkhudi in the province. Culex theileri (40.2%), An. maculipennis s.l. (18.3%), An. superpictus (12.6%), and Cs. longiareolata (12.1%) were the most abundant species found respectively. During this study, An. dthali, An. marteri, An. superpictus, An. turkhudi, and Oc. caspius s.l. were only collected from river edges. Anopehes multicolor however, was found in ground pools, while Cx. perexiguus was encountered from rice fields. Other species were collected from more than one habitat, nonetheless they were most abundant in certain habitats: An. maculipennis s.l. in rice fields (93.0%), Cx. hortensis and Cx. theileri in ground pools (66.7% and 44.4% respectively), Cx. pipiens in artificial ponds (66.7%), and Cs. longiareolata in river edges (50.4%). The association occurrences, percentages, and affinity indices of the collected larvae are discussed.
Key words: Anopheles, Aedes, Culex, Culiseta, Ochlerotatus, larvae, larval habitat, malaria, West Nile virus, affinity index.
Introduction fahan is visited by millions of domestic and for- eign tourists from various parts of the world an- Mosquitoes (Diptera: Culicidae) consist of two nually. These visitors might also carry this mos- subfamilies, 11 tribes, 112 genera, and 3539 species quito borne diseases with them. Some counties in (Harbach 2014). Genus Anopheles Meigen com- Isfahan such as 'Aran and Bidgol', Borkhar, poses of at least 465 species (Harbach 2014). Cer- Khomeinishahr, 'Shahinshahr and Meymeh', and tain mosquito species are involved in the trans- 'Tiran and Krun' are already classified in stratum 2 mission of various arboviral, protozoan and filar- (with the imported cases of malaria and potential ial diseases to humans and domesticated animals, transmission) of the National Malaria Strategy or simply causing nuisance for their biting. The Plan for Malaria Control of Iran (Raeisi et al. 2004). most important mosquito transmitted disease is Another mosquito-borne disease, the West Nile vi- malaria. About 70 Anopheles species are malaria rus, is detected in equines and humans in Isfahan vectors of which around 40 are important vectors through molecular and serological assays (Service 1993). (Ahmadnejad et al. 2011, Chinikar et al. 2012). Malaria is the most important mosquito-borne However, there is no further information about disease in Iran, especially in southern and south- the insect vectors of the virus. Nuisance biting has eastern areas of the country. During 2004–2009 726 also been reported as a problem in Lenjan and malaria cases were reported in Isfahan Province Mobarakeh especially in the areas where human Central Iran, however 96.6% of them were con- habitats and factories are close to rice fields sidered as imported diseases (Ataei et al. 2000, (Mousakazemi et al. 2000). Soleimanifard et al. 2011). As one of the largest The most recent checklist of mosquitoes of and most populated provinces in Central Iran, Is- Iran includes seven genera and 64 species (Azari- The mosquitoes of Central Iran 77
Hamidian 2007b). Oshaghi et al. (2008) identified Province. In 1957 Zulueta et al. determined the Anopheles superpictus Grassi as a complex of three susceptibility level of An. maculipennis to DDT. genotypes (X, Y, and Z). Djadid et al. (2009) found Zaim (1987a) mentioned some aspects of mosquito a new Anopheles species using Polymerase Chain larvae and adult ecology in Kashan County. Reaction (PCR) technique. Naddaf et al. (2010) re- Doroudgar et al. (2000) studied the epidemiology ported An. fluviatilis James species U from Fars of malaria in the same county. In Zarrin-Shahr and Province and were in doubt about the occurrence Mobarakeh areas, Mousakazemi et al. (2000) con- of species (form) V in Iran (Azari-Hamidian ducted the study of mosquito fauna and ecology. 2007b). Species U was also recorded by Mehrava- These limited information are not sufficient as the ran et al. (2011) in southeastern Iran. basis to prevent possible epidemics of mosquito- In Isfahan Province, seven genera and 24 spe- borne diseases. Fauna and ecological data, such as cies of mosquitoes have been reported (Macan larval habitats, species composition, active season, 1950, Dow 1953, Zulueta et al. 1957, Lotfi 1973, and activity peak of mosquito larvae in Isfahan Minar 1981, Zaim 1987a, b, Saebi 1987, Doroudgar Province are very important in integrated vector et al. 2000, Mousakazemi et al. 2000, Sedaghat et management. al. 2003). Among them were ten species of Anophe- les i.e.: An. algeriensis Theobald, An. claviger (Mei- gen), An. dthali Patton, An. maculipennis Meigen, Materials and methods An. marteri Senevet and Prunnelle, An. messeae Study area Falleroni, An. multicolor Combouliu, An. sacharovi Isfahan Province is located in the central plateau of Iran Favre, An. superpictus, and An. turkhudi Liston between 30º42´–34º30´N and 49º40´–55º30´ E with an area (Macan 1950, Dow 1953, Zulueta et al. 1957, Zaim of approximately 107,000 km2. The province is bounded 1987a, Saebi 1987, Doroudgar et al. 2000, Mousa- by Qom and Semnan Provinces in the north, by Markazi, kazemi et al. 2000, Sedaghat et al. 2003). However, Lorestan, 'Chaharmahal and Bakhtiari' Provinces in the there are no recent records of An. algeriensis, An. west, by Yazd Province in the east, and by Fars and dthali, An. marteri, and An. turkhudi. Anopheles sa- 'Kohgiloyeh and Boyerahmad' Provinces in the south (Fig. 1). Isfahan formally includes 23 counties including; charovi and An. maculipennis have also been re- 'Aran and Bidgol', Ardestan, Isfahan, Borkhar, 'Tiran and ported from Maculipennis Group, based on mor- Krun', Chadegan, Khomeinishahr, Khansar, Semirom, phological characters (Zaim 1987a, Saebi 1987, Dehaghan (the lower Semirom), 'Shahinshahr and Mey- Mousakazemi et al. 2000). The latter was also be- meh', Shahreza, Fereydun, Fereydunshahr, Falavarjan ing verified by PCR technique (Sedaghat et al. (the lower Lenjan), Kashan, Golpayegan, Lenjan, Moba- 2003). Zulueta et al. (1957) provided the only re- rakeh, Nain, Najafabad, Natanz, 'Khoor and Biabanak'. cord of An. messeae based on egg chorion pattern. The center of the province, Isfahan City, is almost 1590 m above sea level. The climate ranges from arid and semi- Mousakazemi et al. (2000) reported Aedes vexans arid. In 2006 the province is inhabited by almost 4,560,000 (Meigen). Seven species of Culex Linnaeus have citizen, with main occupations in agriculture, animal been found in the province as follow: Culex horten- husbandry, tourism, business, and industry sectors. sis Ficalbi, Cx. mimeticus Noe, Cx. modestus Ficalbi, Cx. perexiguus Theobald, Cx. pipiens Linnaeus, Cx. territans Walker, and Cx. theileri Theobald (Lotfi 1973, Minar 1981, Zaim 1987a,b, Mousakazemi et al. 2000). Three species of Culiseta Felt are found in Isfahan Province including Cs. longiareolata (Mac- quart), Cs. annulata (Schrank), and Cs. subochrea (Edwards) (Minar 1981; Zaim 1987a, b; Mousaka- zemi et al. 2000). Two species of Ochlerotatus Lynch Arribalzaga, Oc. caspius (Pallas) s.l. and Oc. pulcritarsis (Rondani) have been reported in the province (Zaim 1987a,b, Mousakazemi et al. 2000). Uranotaenia unguiculata Edwards was found by Zaim (1987b). Most conducted studies in Isfahan were re- stricted to mosquito identification. Little informa- Figure 1. Map of Iran, highlighting the position tion on mosquito ecology was recorded in this of Isfahan Province.
78 H. Ladonni et al.
Specimen and data collection An. multicolor, An. superpictus, An. turkhudi, Aedes Larvae sampling was carried out using dipping technique vexans, Culex hortensis, Cx. mimeticus, Cx. per- (WHO 1975) during July–October 2003, and May– exiguus, Cx. pipiens, Cx. territans, Cx. theileri, Cu- September 2004 and 2006. The specimens were collected liseta longiareolata, and Ochlerotatus caspius s.l. This from 20 out of 23 counties (exception: Borkhar, Kashan, and 'Khoor and Biabanak' counties). The frequencies of is the first record of An. dthali, An. marteri, and An. collection were as follow: 'Aran and Bidgol': 8 (Jul–Oct turkhudi in the province. Among 222 collection 2003, Jun 2004), Ardestan: 4 (Aug–Oct 2003), Isfahan: 11 sites, 45 (20.3%) sites were inhabited only by sub- (Aug, Sep 2003, May–Jul 2004), 'Tiran and Krun': 6 (Jul– family Anophelinae, 160 (72.1%) only by Culici- Sep 2003, Aug 2004, Jun 2006), Chadegan: 5 (Jun, Sep nae, and 17 (7.6%) harbored by both subfamilies. 2004, Jun–Jul 2006), Khomeinishahr: 2 (Jun–Jul 2004), In total, anopheline larvae were found in 62 sites Khansar: 4 (Jul–Aug 2004, May–Jun 2006), Semirom: 5 (27.9%), while culicine larvae inhabited 177 (Aug 2003, Jun 2004, Jul 2006), Dehaghan: 1 (Jul 2004), 'Shahinshahr and Meymeh': 1 (Sep 2003), Shahreza: 6 (79.7%) sites. (Aug 2003, Jun, Aug 2004, Jun–Aug 2006); Fereydun: 8 Culex theileri (40.2%), An. maculipennis s.l. (Aug 2003, May–Aug 2006), Fereydunshahr: 8 (Aug–Sep (18.3%), An. superpictus (12.6%), and Cs. longiareo- 2003, Jul 2004, Jul, Sep 2006), Falavarjan: 5 (Jul–Aug 2003, lata (12.1%) were the most prevalent species, re- Jul 2004, Jul 2006), Golpayegan: 2 (Sep 2004, Jul 2006), spectively (Table 1). These species along with Cx. Lenjan: 1 (Aug 2004), Mobarakeh: 5 (Aug 2003, May–Jun perexiguus were widespread in some counties (Ta- 2004, Jul 2006), Nain: 2 (Aug 2003, Jul 2006), Najafabad: 7 ble 2). The association occurrences and percent- (Jun–Aug 2004, Jul 2006), and Natanz: 1 (Jul 2004). Larvae were preserved in lactophenol, while their microscope ages of the collected larvae are shown in Tables 3 slides were prepared using de Faure’s medium. Third and 4, respectively. The months of collection for and 4th instar larvae were identified using the keys of different species is shown in Table 5. The affinity Azari-Hamidian & Harbach (2009). Mosquito name ab- and association indices of mosquito larvae are breviations were following Reinert (2009). Voucher shown in Table 6. During this study, Anopheles specimens were deposited at the Medical Arthropod Mu- dthali, An. marteri, An. superpictus, An. turkhudi, seum in the School of Public Health, Tehran University of and Oc. caspius s.l. were only collected from river Medical Sciences. edges, An. multicolor from ground pools, and Cx. Indices of association between species perexiguus from rice fields. The rest of identified Association indices between pairs of species were calcu- species were found in more than one habitat. lated using two formulae: Fager's index of affinity and as- Anopheles maculipennis s.l. was collected from rice sociation index (Southwood 1978, Silver 2008). Fager's in- fields (93.0%), ground pools (3.5%), and river dex of affinity is defined by IAB=2J/nA+nB where J= num- edges (3.5%). Culex hortensis was discovered from ber of joint occurrences, nA= total number of occurrences ground pools (66.7%) and river edges (33.3%). of species A, and nB= total number of occurrences of spe- cies B. Index significance was analyzed using t-test (at 5% probability level). The total number of occurrences of Table 1. The composition of mosquito larvae collected each species and the number of co-occurrences were in Isfahan Province, July–October 2003, and May– shown in Table 3. Association index is defined by September 2004 and 2006.
I=2[J/A+B–0.5] where J= the number of individuals of both species in samples where they occur together, A = Species n % the total number of individuals of species A in all sam- An. dthali 1 0.1 ples, and B= the total number of individuals of species B An. maculipennis s.l. 304 18.3 in all samples. In this formula, the numbers of individuals An. marteri 45 2.7 within a species s were taken into consideration. The for- An. multicolor 16 1.0 mula has a range of +1 to –1. The total numbers of indi- An. superpictus 209 12.6 viduals of species and the numbers of individuals of co- An. turkhudi 1 0.1 occurring species are shown in Tables 1 and 4. Ae. vexans 1 0.1
Cx. hortensis 111 6.7
Cx. mimeticus 4 0.2 Results Cx. perexiguus 28 1.7 Cx. pipiens 67 4.0 In this investigation, 1659 third- and fourth-instar Cx. territans 2 0.1 larvae including 576 anophelines (34.7%) and 1083 Cx. theileri 667 40.2 culicines (65.3%) were collected from 222 oviposi- Cs. longiareolata 200 12.1 tion sites in Isfahan Province. Fifteen species rep- Oc. caspius s.l. 3 0.1 resenting five genera were identified including; Total 1659 100
Anopheles dthali, An. maculipennis s.l., An. marteri, The mosquitoes of Central Iran 79
Table 2. The distribution of mosquito larvae collected in different counties of Isfahan Province, July–October 2003 and May–September 2004 and 2006.
Aran and Bidgol and Bidgol Aran Ardestan Isfahan Krun Tiran and Chadegan Khomeinishahr Khansar Semirom Dehaghan Shahinshahr Shahreza Fereydun Fereydunshahr Falavarjan Golpayegan Lenjan Mobarakeh Nain Najafabad Natanz County
Species An. dthali ------* ------An. maculipennis s.l. - - * * * * ------* * - * * - * - An. marteri - - - - * ------* ------An. multicolor ------* - - An. superpictus - * - * * - - * - * * * * - * - * * - - An. turkhudi ------* ------Ae. vexans - - - - * ------Cx. hortensis - - - * * - * * - - * * * - - * - - - - Cx. mimeticus - - - - * ------* - - - - - Cx. perexiguus * - * - * - * - - - * - * * - * * - * - Cx. pipiens * - * - * ------* - - * - * - Cx. territans ------* ------Cx. theileri * - * * * * * * * - * * * * * * * * * * Cs. longiareolata * * * * - - * * - - * * ------* - Oc. caspius s.l. ------* - - - - * - -
Table 3. The association occurrences of mosquito larvae collected in Isfahan Province, July–October 2003 and May–September 2004 and 2006 (* - the species was collected alone).
Total occasions An. dthali An. maculipennis marteri An. multicolor An. An. superpictus An. turkhudi Ae. vexans Cx. hortensis Cx. mimeticus perexiguus Cx. pipiens Cx. Cx. territans theileri Cx. Cs. longiareolata Oc. caspius
*
Species s.l. *
s.l.
An. dthali 1 - - - - 1 1 ------An. maculipennis s.l. 31 - - - - 3 - - 1 - 1 1 - 10 - 1 An. marteri 4 - - - - 2 - - - - 1 - - 1 - - An. multicolor* 1 ------An. superpictus 30 1 3 2 - - 1 - 1 1 1 - - 6 - - An. turkhudi 1 1 - - - 1 ------Ae. vexans* 1 ------Cx. hortensis 29 - 1 - - 1 ------1 11 8 1 Cx. mimeticus 3 - - - - 1 ------1 - - Cx. perexiguus 16 - 1 1 - 1 - - - - - 1 - 13 - - Cx. pipiens 14 - 1 ------1 - - 5 3 - Cx. territans 1 ------1 ------Cx. theileri 133 - 10 1 - 6 - - 11 1 13 5 - - 11 1 Cs. longiareolata 32 ------8 - - 3 - 11 - - Oc. caspius s.l. 3 - 1 - - - - - 1 - - - - 1 - -
Culex pipiens was harboring artificial ponds (2.2%). The indices do not show any significant af- (66.7%), rice fields (20%), and ground pools finity between species pairs when they occur to- (13.3%). Culex theileri was found in ground pools gether in larval habitats (at 5% probability level). (44.4%), rice fields (24.5%), river edges (24.2%), ar- tificial ponds (4.6%), and animal foot prints (2.3%), while Cs. longiareolata was collected from river Discussion edges (50.4%), artificial ponds (27.1%), stream edges (12.8%), ground pools (7.5%), and rice fields Five genera and 15 species of mosquitoes were col-
80 H. Ladonni et al.
Table 4. The association percentages of mosquito larvae collected with different species in Isfahan Province, July–October 2003 and May–September 2004 and 2006.
Species association n Abundance (%) An. dthali An. superpictus, An. turkhudi 1 100 An.maculipennis s.l. Alone 183 60.2 Cx. theileri, Cx. perexiguus 59 19.4 Cx. theileri 53 17.4 An. superpictus 4 1.3 Cx. hortensis, Cx. theileri, Oc. caspius s.l. 3 1.0 Cx. pipiens, Cx. theilari 2 0.7 Total 304 100.0 An. marteri Alone 28 62.2 An. superpictus 10 22.2 An. superpictus, Cx. perexiguus, Cx. theileri 7 15.6 Total 45 100.0 An. multicolor Alone 16 100.0 An. superpictus Alone 98 46.9 An. dthali, An. turkhudi 25 12.0 Cx. hortensis, Cx. theileri 24 11.5 Cx. theileri 21 10.0 An. maculipennis s.l. 13 6.2 An. marteri, Cx. perexiguus, Cx. theileri 13 6.2 Cx. mimeticus 13 6.2 An. marteri 2 1.0 Total 209 100.0 An. turkhudi An. dthali, An. superpictus 1 100.0 Ae. vexans Alone 1 100.0 Cx. hortensis Cx. theileri 40 36.1 Alone 33 29.7 Cs. longiareolata 29 26.1 An. maculipennis s.l., Cx. theileri, Oc. caspius s.l. 3 2.7 An. superpictus, Cx. theileri 3 2.7 Cx. theileri, Cs. longiareolata 2 1.8 Cx. territans 1 0.9 Total 111 100.0 Cx. mimeticus An. superpictus 2 50.0 Alone 1 25.0 Cx. theileri 1 25.0 Total 4 100.0 Cx. perexiguus Cx. theileri 22 78.6 Alone 2 7.1 Cx. pipiens 2 7.1 An. maculipennis s.l.,Cx. theileri 1 3.6 An. marteri, An. superpictus, Cx. theileri 1 3.6 Total 28 100.0 Cx. pipiens Cx. perexiguus 26 38.8 Alone 13 19.4
Continued on the next page The mosquitoes of Central Iran 81
Table 4. (continued)
Species association n Abundance (%) Cx. theileri 13 19.4 Cs. longiareolata 9 13.4 An. maculipennis s.l., Cx. theileri 3 4.5 Cx. theileri, Cs. longiareolata 3 4.5 Total 67 100.0 Cx. territans Cx. hortensis 2 100.0 Cx. theileri Alone 371 55.6 Cx. perexiguus 57 8.6 An. maculipennis s.l., Cx. hortensis, Oc. caspius s.l. 41 6.1 Cx. pipiens 38 5.7 An. maculipennis s.l. 33 5.0 Cs. longiareolata 33 5.0 Cx. hortensis 32 4.8 Cx. hortensis, Cs. longiareolata 21 3.1 An. maculipennis s.l., Cx. pipiens 15 2.2 An. maculipennis s.l.,Cx. perexiguus 9 1.4 An. superpictus 9 1.4 Cx. pipiens, Cs. longiareolata 4 0.6 Cx. mimeticus 2 0.3 An. marteri, An. superpictus, Cx. perexiguus 1 0.1 An. superpictus, Cx. hortensis 1 0.1 Total 667 100.0 Cs. longiareolata Alone 74 37.0 Cx. theileri 54 27.0 Cx. hortensis 52 26.0 Cx. hortensis, Cx. theileri 10 5.0 Cx. pipiens 9 4.5 Cx. pipiens, Cx. theileri 1 0.5 Total 200 100.0 Oc. caspius s.l. Alone 2 66.7 An. maculipennis s.l., Cx. hortensis, Cx. theileri 1 33.3 Total 3 100.0
Table 5. The collecting months of mosquito larvae in Isfa- lected and identified in Isfahan Province, includ- han Province, July–October 2003 and May–September ing the first record of An. dthali, An. marteri, and 2004 and 2006. An. turkhudi. One genus, Uranotaenia Lynch Arri- May June July August September October balzaga, and 9 species i.e. An. algeriensis, An. Species claviger, An. messeae, An. sacharovi, Cx. modestus, Cs.
annulata, Cs. subochrea, Oc. pulcritarsis, and Ur. un- An. dthali - - - - * - guiculata which were recorded earlier (Macan An. maculipennis s.l. - * * * * - 1950, Zulueta et al. 1957, Minar 1981, Saebi 1987, An. marteri - - * * * - Zaim 1987a,b, Doroudgar et al. 2000, Mousaka- An. multicolor - - * - - - zemi et al. 2000), were not found in present study. An. superpictus - - * * * * In this investigation we collected six species of An. turkhudi - - - - * - Anopheles, however four species i.e. An. algeriensis, Cx. hortensis - * * * - - Cx. mimeticus - - - - * - An. claviger, An. messeae, An. sacharovi, which were Cx. perexiguus - - * * * - recorded before, were not found. Saebi (1987) did Cx. pipiens * * * * - - not find An. algeriensis in Iran and regarded the Cx. theileri * * * * * - species as very rare eventhough it had been re- Cs. longiareolata * * * * - * corded previously in Fars, Guilan, Isfahan, and Oc. caspius s.l. - - * - - - Kermanshah (Bakhtaran) Provinces. There are few
82 H. Ladonni et al.
Table 6. The affinity and association indices of mosquito larvae in Isfahan Province, July–October 2003 and May–September 2004 and 2006. Lower part shows the index of association. Upper part shows Fager's in- dex of affinity. For bold numbers, see discussion.
An. dthali An. turkhudi Cx. territans Cx. mimeticus Oc. caspius marteri An. pipiens Cx. perexiguus Cx. Cx. hortensis An. superpictus An. maculipennis Cs. longiareolata theileri Cx. Fager's Index of Affinity
s.l.
s.l. Index of Association An. dthali 1 ------0.096 - - - An. turkhudi 1 ------0.064 - - - Cx. territans ------0.066 - - - - Cx. mimeticus ------0.060 - - 0.014 Oc. caspius s.l. ------0.062 - 0.058 - 0.014 An. marteri ------0.100 - 0.117 - - 0.014 Cx. pipiens ------0.066 - - 0.044 0.130 0.068 Cx. perexiguus - - - - - –0.80 –0.42 - 0.043 0.042 - 0.174 Cx. hortensis - - –0.96 - –0.94 - - - 0.033 0.033 0.262 0.135 An. superpictus –0.76 –0.76 - –0.86 - –0.76 - –0.90 –0.84 0.098 - 0.073 An. maculipennis s.l. - - - - –0.98 - –0.98 –0.64 –0.98 –0.94 - 0.123 Cs. longiareolata ------–0.84 - –0.42 - - 0.133 Cx. theileri - - - –0.98 –0.88 –0.98 –0.80 –0.74 –0.64 –0.86 –0.56 –0.72
recent records of An. algeriensis in Iran, which (1950) has reported An. sacharovi from this prov- maybe due to the decreasing of its preferred lar- ince. val habitat such as the reedy swamps (Horsfall Among the collected anophelines, An. dthali, 1955). Anopheles claviger was recorded several An. maculipennis s.l., and An. superpictus are the times in low abundance by Saebi (1987), Zaim proven vectors of malaria in Iran (Hanafi-Bojd et (1987a), and Doroudgar et al. (2000). Zaim (1987a) al. 2011). In view of the greater abundance and collected An. claviger from stream edges and An. wider distribution of two latter species in different multicolor from grassland in Kashan County. To areas of Isfahan (Tables 1 and 2), they might play a date, seven species of the Maculipennis Group more important role in disease transmission. have been recorded in Iran (Azari-Hamidian Mousakazemi et al. (2000) collected two 2007b), however only An. sacharovi is morphologi- anopheline species, An. maculipennis s.l. (2.08% of cally distinguishable from other members of the the whole samples) and An. superpictus (0.16%) group with certainty (Azari-Hamidian & Harbach from rice fields in Lenjan and Mobarakeh areas. In 2009). The most reliable method to distinguish the this study, An. superpictus was only found in river members of the group is the PCR technique, which edges (natural habitat) while A. maculipennis s.l. is still rarely used by the workers. Sedaghat et al. was mostly (93%) encountered in rice fields (arti- (2003) used this method to discover the occurrence ficial habitat). To some extent these results con- of An. maculipennis in Isfahan. Djadid et al. (2007) firmed the observation in Guilan Province, north- also used molecular data in addition to morphol- ern Iran. In this observation Azari-Hamidian ogy to confirm the occurrence of An. messeae in (2011) reported that An. superpictus was only col- Guilan and Mazandaran Provinces (GenBank ac- lected from natural habitats, usually rain pools cession number AY050639). Earlier, Zulueta et al. (67.2%), whereas An. maculipennis s.l. was found in (1957) reported the same species in Isfahan based both natural and artificial habitats, including on egg chorion pattern. In our investigation the 61.7% of man-made habitats of rice fields. Anophe- larvae of Maculipennis Group were distinguished les maculipennis is known as a vector of Setaria in using the mean number of branches of seta 2 on Ardebil Province, northwestern Iran (Azari- the fourth- and fifth- abdominal segments; which Hamidian et al. 2009). is 36.8 for An. sacharovi whereas it is 16.5 for An. Three species of the tribe Aedini i.e.Ae. vexans, maculipennis s.l. (Azari-Hamidian & Harbach Oc. caspius s.l. and Oc. pulcritarsis, were reported 2009). Based on this key, An. sacharovi was not in Isfahan Province (Zaim 1987a,b, Mousakazemi found among the samples, eventhough Macan et al. 2000). In the present study, tree-hole mos- The mosquitoes of Central Iran 83 quito Oc. pulcritarsis was not found. Zaim (1987a) genus, Cs. annulata, and Cs. subochrea, which were collected the species as adults in Kashan of Isfahan discovered earlier in Isfahan (Minar 1981, Zaim Province. The species has also been found in the 1987a,b, Mousakazemi et al. 2000), were not provinces of Golestan, Mazandaran, Semnan (Gut- found. Zaim (1987a) collected Cs. annulata from sevich 1943, Zaim 1987b). Mousakazemi et al. temporary and still fresh water, and permanent- (2000) collected Ae. vexans and Oc. caspius s.l. from slow-running, and to some extent brackish water rice fields in Lenjan and Mobarakeh areas, how- in Kashan County. Mousakezemi et al. (2000) col- ever in present investigation Oc. caspius s.l. was lected Cs. longiareolata from rice fields in Lenjan only found in river edges. and Mobarakeh areas. In present study, Cs. Six species of Culex were found in our investi- longiareolata was found in both artificial (artificial gation (out of seven previously recorded species in ponds and rice fields) (29.3%)and natural habitats the province) (Lotfi 1973, Minar 1981, Zaim 1987a, (ground pools, river edges, and stream edges) b, Mousakazemi et al. 2000). We did not find Cx. (70.7%). Azari-Hamidian (2005) however only modestus, however Cx. theileri was the most abun- found Cs. longiareolata larvae in natural habitat of dant species found (40.2%) of all samples (Table Guilan Province. Gutsevich (1943) reported Cs. 1). Zaim (1987a) collected Cx. pipiens from house annulata in Semnan Province. Kalandadze & ponds and sewage wells and mentioned it as a Kaviladze (1947) found both Cs. annulata and Cs. predominant species in Kashan County especially subochrea (as a subspecies of Cs. annulata) in West in Kashan City. Mousakazemi et al. (2000) col- Azerbaijan and Cs. subochrea in East Azerbaijan lected eight species from rice fields in Lenjan and Province. Minar (1981) and Zaim (1987a) found Cs. Mobarakeh areas where three species Cx. theileri annulata in Isfahan Province. Zaim and Cranston (78.65%), Cx. pipiens (12.83%), and Cx. perexiguus (1986) included Cs. annulata in their checklist and (3.96%) were accounted to more than 90% of the Cs. subochrea in their keys. Zaim (1987b) reported collected samples. We, however only found Cx. Cs. subochrea in 16 provinces of the country. After perexiguus inhabiting rice fields, while Cx. theileri that, Azari-Hamidian et al. (2009) and Saghafipour was more often (70.9%) collected from natural et al. (2012) found the species in Ardebil and Qom habitats (ground pools, river edges, and animal Provinces, respectively. Azari-Hamidian et al. foot prints) in comparison to artificial habitats (2003) and Azari-Hamidian et al. (2009) recorded (29.1%) such as ponds and rice fields). On the con- Cs. annulata in Guilan and Ardebil Provinces, re- trary, Cx. pipiens was mostly collected (86.7%) spectively. In general, Cs. annulata has been re- from man-made habitats (artificial ponds and rice corded in five Iranian provinces by now (Gut- fields), as also observed by Azari-Hamidain sevich 1943, Kalandadze & Kaviladze 1947, Minar (2007a). The present study also verifies previous 1981, Zaim 1987a, Azari-Hamidian et al. 2003, workers (Mousakazemi et al., 2000, Ghavami & Azari-Hamidian et al. 2009) and Cs. subochrea in 18 Ladonni 2005, Abai et al. 2007, Azari-Hamidian et provinces (Zaim 1987, Azari-Hamidian et al. 2009, al. 2009) finding on Cx. theileri as the most abun- Saghafipour et al. 2012). The two species are very dant Culex species in rural areas and relatively similar in the larval stage; however their adults higher altitudes of Iran, such as central and north- can be distinguished easily (Azari-Hamidian & ern Iran, whereas Cx. pipiens is the most prevalent Harbach 2009). The taxonomy and distribution of species in urban areas and lower altitude such as three closely related species, Cs. alaskaensis (Lud- Caspian Sea littoral zone (Zaim 1987a, Azari- low), Cs. annulata, and Cs. subochrea require further Hamidian 2007a). There is little information about study in Iran. the composition percentages of the Pipiens Sub- In this investigation, the most abundant col- group in Southern Iran. The taxonomy, ecology, lected species often occupied their habitats alone and composition of C. pipiens and C quinquefascia- as it was shown in Table 4: An. maculipennis s.l. tus Say and the possibility of gene flow between (60.2%), An. superpictus (46.9%), Cx. theileri (55.6%), them in southern Iran need to be studied exten- and Cs. longiareolata (37.0%). It seems that low pre- sively. Among the collected species, we found cipitation in a province such as Isfahan with arid Cx.theileri, a vector of Dirofilaria in Ardebil Prov- and semiarid climates makes mosquitoes compete ince, northwestern Iran (Azari-Hamidian et al. more in choosing and occupying oviposition sites, 2009). however further study is needed. One species of Culiseta, i.e. Cs. longiareolata, Though, the index of association and Fager's was found in this investigation. Two species of the index of affinity were calculated for all pairs of
84 H. Ladonni et al. species which were collected together (Table 6), sectional serosurvey in equines, 2008–2009. Epidemiology and Infection 139: 1587–1593. not all indices can be analyzed statistically, due to Ataei, B., Labaf Ghasemi, R., Sadri, G.H., Ramezanpour, E., Saeedi, less frequent sampling and rarity in number. A. (2000): A survey on malaria in Isfahan Province during years These include the following species; An. dthali, An. 1986 to 1996. Kowsar Medical Journal 5: 63–67. Azari-Hamidian, S., Joeafshani, M.A., Rassaei A.R., Mosslem, M. marteri, An. turkhudi, Cx. mimeticus, Cx. territans, (2003): Faunistic studies on the genus Culiseta (Diptera: and Oc. caspius s.l. (Tables 1 and 3). That is why Culicidae) in Guilan Province. Journal of Kerman University of both indices of the pair of species An. dthali/An. Medical Sciences 10: 225–233. Azari-Hamidian, S. (2005): Larval habitat characteristics of turkhudi, which were collected on one occasion in- mosquitoes of the genus Culiseta Felt, 1904 (Diptera: Culicidae) cluding one larva, were equal to 1 and show sig- in the Caspian Sea littoral, Iran. Zoology in the Middle East 36: nificant differences (Table 6). When the indices of 59–66. Azari-Hamidian, S. (2007a): Larval habitat characteristics of the pair An. dthali/An. turkhudi are excluded, the mosquitoes of the genus Culex (Diptera: Culicidae) in Guilan remaining indices make more sense. All remaining Province, Iran. Iranian Journal of Arthropod-Borne Diseases 1: results for the index of association are negative, 9–20. Azari-Hamidian, S. (2007b): Checklist of Iranian mosquitoes hence they do not show any significant differences (Diptera: Culicidae). Journal of Vector Ecology 32(2): 235–242. (association). Also, t was less than 1.645 (at 5% Azari-Hamidian, S., Harbach, R.E. (2009): Keys to the adult females probability level) for all pairs of species for which and fourth-instar larvae of the mosquitoes of Iran (Diptera: Culicidae). Zootaxa 2078: 1–33. Fager's index of affinity was calculated (no affin- Azari-Hamidian, S., Yaghoobi-Ershadi, M.R., Javadian, E., Abai, ity). M.R., Mobedi, I., Linton, Y.-M., Harbach, R.E. (2009): There are many vectors of human and domes- Distribution and ecology of mosquitoes in a focus of dirofilariasis in northwestern Iran, with the first finding of ticated animal pathogens such as An. dthali, An. filarial larvae in naturally infected local mosquitoes. Medical maculipennis, An. superpictus, Ae. vexans, Cx. per- and Veterinary Entomology 23: 111–121. exiguus, Cx. pipiens, Cx. theileri, and Oc. caspius s.l. Azari-Hamidian, S. (2011): Larval habitat characteristics of the genus Anopheles (Diptera: Culicidae) and a checklist of (Horsfall 1955, Harbach 1988, Azari-Hamidian et mosquitoes in Guilan Province, northern Iran. Iranian Journal of al. 2009, Hanafi-Bojd et al. 2011) among the mos- Arthropod-Borne Diseases 5: 37–53. quitoes of the province that different aspects of Chinikar, S., Javadi, A., Ataei, B., Shakeri, H., Moradi, M., Mostafavi, E., Ghiasi, S.M. (2012): Detection of West Nile virus their ecology such as active season, host prefer- genome and specific antibodies in Iranian encephalitis patients. ence, and population dynamics need to be studied Epidemiology and Infection 140: 1525–1529. extensively. Djadid, N.D., Gholizadeh, S., Tafsiri, E., Romi, R., Gordeev, M., Zakeri, S. (2007): Molecular identification of Palearctic members of Anopheles maculipennis in northern Iran. Malaria Journal 6: 6– 15. Djadid, N.D., Jazayeri, H., Gholizadeh, S., Rad, Sh.P., Zakeri, S. Acknowledgements. We thank Dr. K. 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