Culex Pipiens S.S

Mosquito Surveillance and Management in Turkey

Filiz GÜNAY and Bülent Alten

Native and non-native vector management in the Eastern Mediterranean and Middle East (EMME) 18-20 April 2018, Cyprus MOSQUITO BORN DISEASES IN TURKEY

1970 2013 Vector species in Turkey Widespread in the 1994 - 2012 2017 Number of cases Number of Anopheles sacharovi Number of cases Number of Malaria past cases Anopheles superpictus 170.000 84.345 - 5 cases 101 285(MoH)

West Nile Vector species in Turkey Culex pipiens s.s. Difﬁcult to control Egean, Mediterranean, Blood samples from Human Central, South East, Birds Horse Dog and Cow Encephelitis Culex quinquefasciatus Anatolia Culex perexiguus

Vector species in the world Aedes aegypti Rearly seen in Turkey Central Anatolia antigens in Dengue Aedes albopictus serum of animals Past studies on Mosquito Surveillance in Turkey

Parrish 1959 Ramsdale et.al. 2001 Aldemir et.al. 2009 SOVE Simsek et.al. 2011 1 Anopheles algeriensis Anopheles algeriensis 2 Anopheles claviger Anopheles claviger § D Parrish 1959 3 Anopheles hyrcanus Anopheles hyrcanus 4 Anopheles maculipennis ss Anopheles maculipennis ss 5 Anopheles messeae The Mosquitoes of Turkey 6 Anopheles sacharovi Anopheles sacharovi 7 Anopheles melanoon Anopheles (subalpinus) melanoon Anopheles melanoon 52 Tür; Anopheles (13), Aedes (17), Culex (16), Culiseta (4), 8 Anopheles marteri Anopheles marteri 9 Anopheles plumbeus Anopheles plumbeus Uranotaenia (1), Orthopodomyia (1). 10 Anopheles pulcherrimus 11 Anopheles superpictus Anopheles superpictus 12 Acartomyia phoeniciae 13 Acartomyia zammitii § CD Ramsdale, B Alten, SS Çağlar and N Özer 2001 14 Aedes cinereus 15 Aedes vexans Aedimorphus vexans 16 Aedes geniculata Dahliana geniculata A Revised, Annotated Checklist of the Mosquitoes (Diptera, 17 Aedes echinus Dahliana echinus 18 Aedes annulipes Listede yer verilmemiş Culicidae) of Turkey 19 Aedes caspius Ochlerotatus caspius 20 Aedes communis Ochlerotatus communis 48 Tür; Anopheles (10), Aedes (3), Ochlerotatus (15), Culex (13), 21 Aedes detritus Ochlerotatus detritus 22 Aedes dorsalis Ochlerotatus dorsalis Culiseta (4), Coquillettidia (1), Uranotaenia (1), Orthopodomyia (1). 23 Aedes excrucians Ochlerotatus excrucians 24 Aedes flavescens Ochlerotatus flavescens 25 Ochlerotatus cataphylla 26 Ochlerotatus cyprius § A Aldemir, F Erdem, B Demirci, H Bedir, E Koç 2009 27 Ochlerotatus leucomelas 28 Ochlerotatus pullatus 29 Ochlerotatus punctor Species Composition and Seasonal Dynamics of Mosquito 30 Aedes lepidonotus Ochlerotatus lepidonotus 31 Aedes nigrocanus Ochleotatus nigrocanus Larvae (Diptera: Culicidae) in Kars Plateau and Six New 32 Aedes pulcritarsis Ochlerotatus pulcritarsis 33 Aedes refiki Ochlerotatus refiki Records for Turkey 34 Aedes rusticus Ochlerotatus rusticus 35 Stegomyia cretina Aedes cataphylla, Aedes cyprius, Aedes leucomelas, Aedes punctor, 36 Culex deserticola Culex deserticola 37 Culex hortensis Culex hortensis Aedes pullatus, Culiseta alaskaensis 38 Culex laticinctus Culex laticinctus 39 Culex martinii Culex martinii 40 Culex mimeticus Culex mimeticus 41 Culex modestus Culex modestus § FM Şimşek, C Ülger, MM Akıner, F Günerkan, SI Cihangir, F 42 Culex perexiguus 43 Culex pusillus Culex pusillus Bardakçı 2011 44 Culex theileri Culex theileri 45 Culex quinquefasciatus Hatalı Kayıt Olarak Belirtilmiş Mosquito Species in Southern Turkey 46 Culex pipiens ss Culex pipiens ss 47 Culex torrentium Culex torrentium 48 Culex tritaeniorhynchus Culex tritaeniorhynchus Anopheles melanoon, Culiseta subochrea 49 Culex territans 50 Culiseta alaskaensis 51 Culiseta annulata Culiseta annulata Parrish 1959 Ramsdale et.al. 2001 52 Culiseta subochrea 59 Anopheles sinensis Hatalı Kayıt 53 Culiseta fumipennis Culiseta fumipennis 60 Anopheles multicolor Günümüzde Mevcut Değil 54 Culiseta longiaerolata Culiseta longiaerolata 61 Anopheles sergentii Günümüzde Mevcut Değil 62 Aedes aegypti Günümüzde Mevcut Değil 55 Culiseta morsitans Culiseta morsitans 63 Aedes mariae Hatalı Kayıt 56 Mansonia richiardii Coquillettidia richiardii 64 Culex apicalis Hatalı Kayıt 65 Culex adairi Hatalı Kayıt 57 Orthopodomyia pulcripalpis Orthopodomyia pulcripalpis 66 Culex univittatus Hatalı Kayıt 58 Uranotaenia unguiculata Uranotaenia unguiculata 67 Culex vagans Hatalı Kayıt

Genel Bilgiler Gereç ve Yöntem Bulgular ve Tartışma Sonuçlar Difﬁculties on morphological identiﬁcation

• Samples can be damaged Classical Taxonomy DNA Barcoding • Seta and scales fall off, colours fade away Morphological Molecular • Recognition requires experience and Characters Markers expertise

• Phenotypic plasticity and morphological variation of neotype and alloneotype NEW TAXONOMY MOLECULAR IDENTIFICATION

§ DNA barcoding: cytochrome oxidase subunit I (COI) - Neighbor joining (NJ) method using Kimura 2 parameter

§ Barcode of Life Data System, BOLD Natural History Museum in London and Washington DC

§ Reliable data to compare sequences was created § Molecular evolution is slow on mtDNA and distance within two species is rarely over 2%. SAMPLING SITES

2010-2017: 34 Cities from all over Turkey Standard Method

COI-ITS2-EF-ACE 1959-2011: 54 taxa 2010-2017: 63 taxa;

Anopheles 10 Anopheles 13 taxa (+3) Aedes 23 Aedes 24 spp. (+1) Culex 13 Culex 16 taxa (+3) Culiseta 5 Culiseta 6 spp. (+1) Coquillettidia 1 Coquillettidia 2 spp. (+1) Uranotaenia 1 Uranotaenia 1 sp. Orthopodomyia 1 Orthopodomyia 1 sp. 86 An maculipennis (35 haplotip)

71 An MBI-36 (34 haplotip) 65 69 An melanoon (7 haplotip) 100 96 An messeae (8 haplotip) 98 70 An sacharovi (16 haplotip) 100 82 An hyrcanus (4 haplotip)

100 15 An pseudopictus (7 haplotip) 96 100 99 An algeriensis (4 haplotip) 94 An algeriensis (3 haplotip) 82 100 An plumbeus Haplotip 32 An superpictus (11 haplotip) 100 2 An claviger (19 haplotip) 100 100 Ae geniculatus (4 haplotip) Ae rusticus Haplotip 100 30 Ae detritus (4 haplotip) 100 Ae leucomelas (5 haplotip) 27 33 Ae communis Haplotip

34 96 Ae punctor (2 haplotip) 41 100 100 Ae pullatus (2 haplotip)

29 Ae flavescens Haplotip1 99 Ae annulipes (4 haplotip) 16 100 99 Ae pulcritarsis (4 haplotip) 99 6 Ae phoeniciae (2 haplotip) 57 Ae caspius Haplotip9

98 77 Ae caspius Haplotip11 99 Ae zammitii Haplotip 43 Ae caspius Haplotip68

Ae caspius (68 haplotip) 36

100 Ae vexans (35 haplotip)

30 Ae albopictus Haplotip 100 Cs longiareolata (13 haplotip) 100 100 Cs annulata (2 haplotip) Cs annulata benzeri Haplotip 100 Cx hortensis (12 haplotip) 48 100 Cx sp1 (2 haplotip)

99 Cx impudicus (3 haplotip) 100100 Cx tritaeniorhynchus (8 haplotip) 3 30 100 Cx modestus (7 haplotip) 82 Cx torrentium (7 haplotip) 53 Cx pipiens f molestus Haplotip 59 99 99 Cx quinquefasciatus Haplotip 7 69 Cx pipiens ss Haplotip 35 Cx laticinctus Haplotip

99 Cx theileri (21 haplotip) 44 78 Cx perexiguus (5 haplotip) 10 100 Cx mimeticus (3 haplotip) 100 Ur unguiculata (2 haplotip) 4 100 Cq buxtoni Haplotip

100 Cq richiardii (3 haplotip) 100

0.02 Anophelinae Subfamily

87 An.An. MBI-36 MBI-36 (35 (35 Haplotip) Haplotypes) 58

86 § 24 cities 1.794 samples - An.An. maculipennis maculipenis s.s. s.s. (30 (30 Haplotip) Haplotypes) 62 90 345 mosquitoes barcoded An.An. melanoon melanoon (1 (1Haplotip) Haplotype) 100 An.An. messeae messeae (8 (8Haplotip) Haplotypes) 99 62 An.An. sacharovi sacharovi (16 (16 Haplotip) Haplotypes) 100 80 An.An. hyrcanus hyrcanus (4 (4Haplotip) Haplotypes) 39 100 An.An. pseudopictus pseudopictus (7 Haplotip)(7 Haplotypes) 96 36 An.An. marteri marteri (3 (3örnek samples BOLD) BOLD) 100 § 10 Anopheles species + 4

An.An. claviger claviger (19 (19 Haplotip) Haplotypes) 100 new taxon An. An.pulcherrimus pulcherrimus (1 sample (1 örnek GenBank) GenBank)

83 An.An. superpictus superpictus (11 (11 Haplotip) Haplotypes) 100 An.An. plumbeus plumbeus (1 (1 Haplotip) Haplotype) 100 82 An.An. algeriensis algeriensis benzeri like (3 (3 Haplotypes) Haplotip)

99 An.An. algeriensis algeriensis (4 (4Haplotip) Haplotypes) 100

0.02 Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) algeriensis, Theobald, 1903

§ 40 samples are An. algeriensis, 16 of them barcoded

§ 6 of them are 5% different than the other 10

samples !

An. algeriensis Haplotype 1

An. algeriensis Haplotype 2

An. algeriensis Haplotype 3

An. algeriensis Haplotype 4

An. algeriensis like Haplotype 1

An. algeriensis like Haplotype 2

An. algeriensis like Haplotype 3 Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) claviger, Meigen, 1804

§ Anopheles claviger s.l. 17 cities 74 samples, 28 were barcoded § None of them are Anopheles petragnani with 6.8% distance

Morphological Identification Morphological and Molecular Identification Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) hyrcanus, Pallas, 1771

§ 4th tarsal segment

§ 200 samples, 26 were barcoded

§ 1,4 % genetic distance

Anopheles hyrcanus (a) Anopheles pseudopictus (b)

Morphological and Molecular Identification Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) hyrcanus, Pallas, 1771

Anopheles hyrcanus (a) Anopheles pseudopictus (b)

FJ210895.1|An. hyrcanus AnophelesFJ210895.1 GenBank hyrcanus T A T A A T T A T A A C A C C C [562] 99 AnAn. hyrcanus hyrcanus Haplotip1 Haplotype 1 ...... [562] 63 AnAn. hyrcanus hyrcanus Haplotip2 Haplotype 2 ...... G . . . [562] 34 AnAn. hyrcanus hyrcanus Haplotip4Haplotype 4 ...... G . . . [562] 50 AnAn. hyrcanus hyrcanus Haplotip3Haplotype 3 ...... T . [562] AnAn. pseudopictus pseudopictus HaplotypeHaplotip2 2 A . C . . C . . . . G T . . . A [562] AnAn. pseudopictus pseudopictus Haplotip1Haplotype 1 A . C . G C . . . . G T . . . A [562] 54 AnAn. pseudopictus pseudopictus Haplotip3Haplotype 3 A G C . . C . . . . G T . . . A [562] 42 AnAn. pseudopictus pseudopictus HaplotypeHaplotip4 4 A G C . . C A . . . G T . . . A [562] 38 7 AnAn. pseudopictus pseudopictus Haplotip9Haplotype 9 A . C . . C A . . . G T . . . A [562] AnAn. pseudopictus pseudopictus Haplotip7Haplotype 7 A . C G . C . G . . G T . . . A [562]

13 63 AnAn. pseudopictus pseudopictus Haplotip5Haplotype 5 A . C . . C . . . . G T . A . A [562] AnAn. pseudopictus pseudopictus Haplotip8Haplotype 8 A . C . . C . . . . G T . T . A [562] 11 AnAn. pseudopictus pseudopictus HaplotypeHaplotip6 6 A . C . . C . . C G G T . . . A [562] Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) maculipennis, Meigen, 1818 species complex

In Palearctic: § An. artemievi Gordeev, 2005; § An. atroparvus van Thiel, 1927; § An. beklemishevi Stegnii ve Kabanova, 1976; § An. daciae Nicolescu, 2004; § An. labranchiae Falleroni, 1926; § An. maculipennis s.s. Meigen, 1818; § An. martinius Shingarev, 1926; § An. melanoon Hackett, 1934; § An. messeae Falleroni, 1933; § An. persiensis Linton, 2003; § An. sacharovi Favre, 1903

§ 11 species Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) maculipennis, Meigen, 1818 species complex

§ 11 species Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) maculipennis, Meigen, 1818 species complex

§ An. maculipennis s.s. (n= 172), An. messeae (n= 11), An. melanoon (n= 3) and An. sacharovi (n= 42)

47 An.An. maculipennis maculipenis s.s.s.s. (30 (30 Haplotypes Haplotip ve and 12 12 örnek samples GenBank) from GenBank)

34 An. MBI-36 (35 Haplotip) 49 An. MBI-36 (35 Haplotypes)

An.An. melanoon melanoon (1(1 Haplotype)Haplotip)

48 LK054515.1|An. atroparvus Anopheles LK054515.1 atroparvus GenBank

34 An.An. labranchiae labranchiae (9 samples örnek GenBank) from GenBank) 79 99 An.An. beklemishevi beklemishevi (10 samples örnek GenBank) from GenBank) 36 86 An.An. messeae messeae (7(7 HaplotipHaplotypes ve and 1 örnek 1 sample GenBank) from GenBank)

39 An.An. daciae daciae (9 (9 örneksamples GenBank) from GenBank) 30

An.An. sacharovi sacharovi (15 (15 HaplotipHaplotypes ve and4 örnek 4 samples GenBank) from 100 GenBank)

0.02 Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) maculipennis, Meigen, 1818 species complex

§ An. maculipennis s.l. Samples from 2013 Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) maculipennis, Meigen, 1818 species complex

§ 22 cities 1.382 samples of An. maculipennis s.l. – 228 were barcoded Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) maculipennis s.s. Meigen, 1818

§ An. maculipennis s.l. 75 % are An. maculipennis s.s. And they are forming 2 groups: 1,4 % genetic distance § Genetic distance between An. messeae and An. daciae is 0.7%!!! § NEW SPECIES?

84 An.An. MBI-36 MBI-36 (35 (35 Haplotip) Haplotypes) 57

59 An.An. maculipennis maculipenis s.s.s.s. (30(30 Haplotypes) Haplotip) 61

An.An. melanoon melanoon (1 (1 Haplotip)Haplotype)

An.An. messeae messeae (8 (8 Haplotip) Haplotypes) 99 An.An. sacharovi sacharovi (16 (16 Haplotip) Haplotypes) 99

0.02 Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) maculipennis s.s. Meigen, 1818

Anopheles MBI-36 Anopheles maculipennis s.s. Anopheles messeae Anopheles melanoon Anopheles sacharovi

84 An.An. MBI-36 MBI-36 (35 (35Haplotip) Haplotypes) 57

59 An.An. maculipennis maculipenis s.s. s.s. (30 (30 Haplotip) Haplotypes) 61

An.An. melanoon melanoon (1 Haplotip) (1 Haplotype)

An.An. messeae messeae (8 Haplotip) (8 Haplotypes) 99 An.An. sacharovi sacharovi (16 (16Haplotip) Haplotypes) 99

0.02 Anopheles Genus Anopheles Subgenus

Anopheles (Anopheles) maculipennis s.s. Meigen, 1818

84 An.An.An. MBI-36 MBI-36MBI-36 (35 (35(35Haplotip) Haplotypes)Haplotypes) 57

59 An.An. maculipennis maculipenis s.s. s.s. (30 (30 Haplotip) Haplotypes) 61

An.An. melanoon melanoon (1 Haplotip) (1 Haplotype)

An.An. messeae messeae (8 Haplotip) (8 Haplotypes) 99 An.An. sacharovi sacharovi (16 (16Haplotip) Haplotypes) 99

0.02 Anopheles Genus Anopheles Subgenus

Anopheles (Cellia) superpictus, Grassi, 1899 § Adana, Aydın and Erzincan

100 3 cities – South, West and An.An. superpictus superpictus (11 (11Haplotip Haplotypes ve 5 örnek and GenBank) 5 samples from GenBank) East Anatolia An.An. pulcherrimus pulcherrimus (3 örnek(3 samples GenBank) from GenBank) 100 An.An. stephensi stephensi (3 örnek(3 samples GenBank) from GenBank) 100 § 2nd important malaria

0.02 vector

Anopheles (Anopheles) plumbeus, Stephens, 1828

An plumbeus (1 Haplotip)

KM258215.1| Anopheles plumbeus

KM258216.1| Anopheles plumbeus

KM258217.1| Anopheles plumbeus

KM258218.1| Anopheles plumbeus

KM258219.1| Anopheles plumbeus

An claviger (1 Haplotip)

0.02 Culicinae Subfamily; Aedini Tribe

89 33 Ae.Ae zammitii. zammitii (3 (3 Haplotip) Haplotypes)

Ae.Ae phoeniciae. phoeniciae (2 (2 Haplotip) Haplotypes) 99 48 § 20 cities 1968 Aedes samples – 296

Ae.Ae caspius. caspius (66 (66 Haplotip) Haplotypes) barcoded 83 55

Ae.Ae dorsalis. dorsalis (4 (4 örnek samples GenBank) from GenBank) 89 3 Ar.Ae pulchritarsis. pulchritarsis (4 (4 Haplotip) Haplotypes) 100 § Aedes albopictus 100 Ae. pullatus (2 Haplotip) 100 new record for the Ae. leucomelas (6 Haplotip) 26 22 country 51 Ae. communis (1 Haplotip) 96 Ae. punctor (8 Haplotip) 100 100 Ae. detritus (4 Haplotip) 100 § Aedes aegypti and 45 Ae.Ae cyprius. cyprius (2 (2 örnek samples GenBank) from GenBank) 61 99 Aedes albopictus in 45 Ae. flavescens (2 Haplotip) 96 Ae. annulipes (4 Haplotip) North East Turkey 99 Ae. echinusechinus (1 (1 sample örnek fromGenBank) GenBank) 92 Ae. geniculatus (4 Haplotip) 100 Ae. rusticus (1 Haplotip) 100 64 Ae.Ae cinereus. cinereus (4 (4 örnek samples GenBank) from GenBank) § Presence of Ae. 69 Ae. albopictus (1 Haplotip) annulipes, Ae. Ae.Ae cretinus. cretinus (3 (3 örnek samples BOLD) from BOLD) 100 89 leucomelas, Ae. Ae.vexansAe. vexans (35 (35 Haplotip) Haplotypes) and 100 pullatus Ae. punctor proven.

0.02 Culicinae Subfamily; Aedini Tribe

Stegomyia Subgenus

Ae albopictus (1 Haplotip)

0.02 Culicinae Subfamily; Aedini Tribe

Aedes albopictus in Aedes aegypti and North West Turkey Aedes albopictus in - 2011- North East Turkey 2015. Akiner et al. 2016 2080

Ae. aegypti 2050 2017 Ae. albopictus 2017 Ae. aegypti

2013

Schaffner et al. 2014, Lancet Culicinae Subfamily; Aedini Tribe

Aedes and Aedimorphus Subgenus

§ Aedes (Aedes) cinereus, Meigen, 1818

§ Aedes (Aedimorphus) vexans, Meigen, 1830

100 Ae.Ae . vexansvexans (35(35 Haplotypes) Haplotip)

Ae.Ae . cinereuscinereus (3(3 samples örnek GenBank)from GenBank)

0.01 Culicinae Subfamily; Culicini Tribe

§ Culex (Barraudius) modestus, Ficalbi, 1889

§ Culex (Barraudius) pusillus, Macquart, 1850

§ Culex (Culex) laticinctus, Edwards, 1913

§ Culex (Culex) mimeticus, Noe, 1899

§ Culex (Culex) perexiguus, Theobald, 1903

§ Culex (Culex) pipiens ss, Linnaeus, 1758

§ Culex (Culex) pipiens f. molestus, Forskal 1775

§ Culex (Culex) quinquefasciatus, Say, 1823 100 31 Cx. tritaeniorhynchus (8 Haplotip) § Culex (Culex) theileri, Theobald, 1903 Cx. mimeticus (3 Haplotip) 100 14 80 Cx. pipiens (1 Haplotip) § 31 cities, 2058 Culex § Culex (Culex) torrentium, Martini, 1925 99 Cx. quinquefasciatus (1 Haplotip) samples – 247 23 Cx. pipiens f. molestus (1 Haplotip) § Culex (Culex) tritaeniorhynchus, Giles, 1901 94 barcoded Cx. torrentium (7 Haplotip) 19 66 Cx. modestus (7 Haplotip) § Culex (Maillotia) deserticola, Kirkpatrick, 1924 100 Cx. laticinctus (1 Haplotip) § Barcodes represent 13 98 § Culex (Maillotia) hortensis Ficalbi, 1889 99 Cx. theileri (19 Haplotip) taxa 91 Cx. perexiguus (4 Haplotip) § Culex (Neoculex) europaeus, da Cunha Ramos, 100 100 § 66 Cx. impudicus (5 Haplotip) Cx. impudicus ve Cx. Ribeiro ve Harrison, 2003 100 Cx. territans (3 örnek GenBank) quinquefasciatus are 99 Cx. sp 1 (2 Haplotip) new records for 100 § Culex (Neoculex) impudicus, Ficalbi, 1890 Cx. hortensis (12 Haplotip) Turkey 100

§ Culex (Neoculex) martinii, Medschid, 1930 0.02 Culicini Tribe; Culex Genus

Neoculex Subgenus

§ Culex (Neoculex) europaeus, da Cunha Ramos, Ribeiro ve Harrison, 2003

§ Culex (Neoculex) impudicus, Ficalbi, 1890

§ Culex (Neoculex) martinii, Medschid, 1930

Cx. sp. 1

Cx. impudicus

Cx. territans ! Culicini Tribe; Culex Genus

Culex Subgenus Culex pipiens species complex and Culex torrentium

!Culex&pipiens&s.s.& &Culex&quinquefasciatus& &Culex&pipiens&f.&molestus& &Culex&torren4um& Culicini Tribe; Culex Genus

Culex Subgenus

Cx. pipiens s.s. (1 Haplotip) GA [316] 80

100 Cx. quinquefasciatus (1 Haplotip) .G [316]

Cx. pipiens f molestus (1 Haplotip) A. [316]

Cx. torrentium (7 Haplotip)

0.02 New records for Turkey:! Ae. albopictus, ! An. hyrcanus var. pseudopictusCq. buxtoni, ! Cx. impudicus

Cq. buxtoni, Cx. hortensis, Cx. laticinctus ve Cx. perexiguus were barcoded for the ﬁrst time

We aproved the presence of Ae. annulipes, Ae. leucomelas, Ae. pullatus, Ae. punctor, An. messeae, Cx. quinquefasciatus and Cx. pipiens f. molestus molecularly ARBOVIRUSES ON THE SYRIAN BORDER Kum sineği Hotspots: tür+ IRD, Montpellier, France March 10th, 2011 parazit+virus Syria Project

28441 Specimens (Morphology)

28203 Specimens (Virus screening)

238 Specimens (DNA barcoding)

8 pools ( Virus +) ARBOVIRUSES ON THE SYRIAN BORDER ARBOVIRUSES ON THE SYRIAN BORDER ARBOVIRUSES ON THE SYRIAN BORDER

Environment Ground Use No.

Other Bamboo grove Urban Urban use (road, buildings) Village Pasture Coltivated field Orchard Grove/Plantation Beach Agriculture Coniferous forest None (wild) No-info Prairie 0 200 400 600 800 1000 1200 Savana Scrub/brush Island Salt marsh Terrain Mangrove Swamp Cloud forest no-info Rain forest valley Plain Desert plateau Deciduous forest plain Evergreen forest mountain 0 200 400 600 800 hill

0 100 200 300 400 500 600 700 800 ARBOVIRUSES ON THE SYRIAN BORDER

Sky No. Shade

Heavy rain

Light Rain no-info

Fog heavy Mist

Partially cloudy partial Cloudy

Overcast none Clear

0 200 400 600 800 1000 1200 1400 0 200 400 600 800

wind

no-info strong gusts light none 0 100 200 300 400 500 600 ARBOVIRUSES ON THE SYRIAN BORDER

200

180

160

140

120

100

0 ARBOVIRUSES ON THE SYRIAN BORDER

Osmaniye:

Aedes caspius Anopheles claviger Anopheles maculipennis s.l. Anopheles sachorovi Culex perexiguus Culex pipiens s.l. Culex tritaeniorhynchus

Adana:

Aedes caspius Anopheles claviger Hatay: Anopheles maculipennis s.l. Kahramanmaraş: Anopheles sachorovi Aedes caspius Culex perexiguus Aedes dorsalis Culex pipiens sl Culex pipiens s.l. Culiseta longiareolata Culex theileri Culex tritaeniorhynchus Culex pipiens sl Uranotaenia unguiculata Culex theileri ARBOVIRUSES ON THE SYRIAN BORDER

Çanakkale:

Aedes caspius Culiseta longiareolata Culex pipiens s.l. Culex theileri

Balıkesir:

Culex pipiens sl Culex theileri

İzmir:

Aedes caspius Anopheles superpictus Culiseta longiareolata Culex pipiens sl Culex theileri ARBOVIRUSES ON THE SYRIAN BORDER

Kiklareli: Edirne: Tekirdag: Aedes caspius Aedes caspius Aedes caspius Aedes pulchritarsis Anopheles claviger Aedes pulchritarsis Anopheles maculipennis sl Anopheles maculipennis sl Anopheles claviger Culiseta annulata Coquillettidia richiardii Anopheles maculipennis sl Culiseta longiareolata Culex pipiens sl Culiseta longiareolata Culex pipiens sl Culex theileri Culex pipiens sl Culex theileri Uranotaenia unguiculata virus detection Pool code year Morphological ID Molecular ID province Virus detected

TK599-1 2014 Culiseta annulata Culiseta annulata Kirklareli Chikungunya (NW) virus TK614-3A 2014 Culex pipiens s.l. Culex pipiens s.s. Kirklareli Chikungunya (NW) virus TK610-12A 2014 Culex pipiens s.l. Culex pipiens s.s. Tekirdag Chikungunya (NW) virus TK587-1 2014 Anopheles maculipennis Anopheles maculipennis Kirklareli Chikungunya s.l. s.s. (NW) virus TR138-408-160 2015 Aedes caspius Aedes caspius Kirklareli Chikungunya 8R (NW) virus TK579-1A 2014 Anopheles maculipennis Anopheles maculipennis Kirklareli Chikungunya s.l. s.s. (NW) virus TR139-23-46 2015 Culex pipiens s.l. Culex pipiens s.s. Kirklareli West Nile virus (NW) TR013-55-59 2015 Anopheles maculipennis Anopheles sacharovi Adana (SE) Chikungunya s.l. virus q The presence of Chikungunya virus in the country was recorded for the ﬁrst time q The ongoing circulation of West Nile virus was revealed q CHICK virus has been found in inefﬁcient vector species but recent studies have shown the distribution of Aedes albopictus and Aedes aegypti is expanding in the North East of Turkey, which increases the risk of transmission q Executing a mosquito DNA barcode reference database is helpful to estimate the primary vector species in the critical areas