Öz
Amaç
Bu çalışmada İzmir’in farklı ilçelerinde yayılış gösteren Culicoides türlerinin belirlenmesi, moleküler karakterizasyonlarının yapılması ve kanatlı haemosporidian parazitleri yönünden vektörlük potansiyellerinin araştırılması amaçlanmıştır.
Yöntemler
Çalışma materyalini, 2016 yılının Mayıs-Ağustos ayları arasında Bergama, Ödemiş, Kemalpaşa ve Foça ilçelerinden toplanan toplam 800 adet dişi Culicoides örneği oluşturmuştur. Culicoides örneklerinin morfolojik teşhislerini takiben belirlenen her türe ait örnekler moleküler analizlere dahil edilmiştir. Örneklere ait genomik DNA izolatlarının mt-COI gen bölgesi polimeraz zincir reaksiyonunda (PZR) amplifiye edilmiş ve sonraki basamakta amplifikasyon ürünlerinin sekans analizleri gerçekleştirilerek moleküler karakterizasyonları ve filogenetik analizleri yapılmıştır. Tür ve lokasyonuna göre ayrılan örneklerin baş/toraks (BTH) ve abdomenlerinden ayrı havuzlar oluşturularak elde edilen gDNA izolatlarında haemosporidian DNA’sının varlığı nested PZR ile araştırılmış ve tespit edilen parazit nesillerinin sekans analizleri ile moleküler karakterizasyonları yapılmıştır.
Bulgular
Morfolojik teşhis sonuçlarına göre araştırma bölgelerinde C. circumscriptus (%39,4) ve C. imicola’nın (%33,8) en yaygın türler olduğu bunu sırasıyla Culicoides sp. (ERU-Izm-Culi1) (%9,1), C. nubeculosus kompleks (%7,6), C. obsoletus (%4,3), C. gejgelensis (%2,3), C. punctatus (%1,9) ve C. newsteadi’nin (%1,8) izlediği saptanmıştır. Belirlenen türlere ait mt-COI sekansları arasında 18 farklı haplotipi ortaya koyan 175 polimorfik bölge saptanmış olup haplotip çeşitliliği en yüksek C. circumscriptus, C. punctatus ve C. newsteadi’de belirlenmiştir. Filogenetik analizlerde belirlenen Culicoides türlerine ait haplotipler üç major küme içerisinde monofiletik olarak gruplanmıştır. Culicoides circumscriptus BTH’ye ait genomik DNA izolatlarında Haemoproteus sp. GAGLA05 ve H. minutus TURDUS2 nesillerinin varlığı saptanmış ve bu türün araştırma yöresinde ilgili Haemoproteus nesillerinin muhtemel potansiyel vektörü olabileceği ortaya çıkarılmıştır.
Sonuç
Bu çalışma ile İzmir yöresinde yaygınlık gösteren Culicoides türleri entegre morfolojik ve moleküler teşhis yaklaşımlarıyla belirlenerek bu önemli sineklerin moleküler epidemiyolojisi açısından özgün veriler sağlanmıştır. Ayrıca çalışma sonuçları İzmir yöresinde C. circumscriptus’un Haemoproteus nesillerinin nakli açısından potansiyel bir öneme sahip olduğuna dair kanıtlar sağlamıştır.
Anahtar Kelimeler: Culicoides, moleküler karakterizasyon, kanatlı Haemosporidian parazitler, vektör potansiyeli, İzmir
Kaynakça
Mellor PS, Boorman J, Baylis M.Culicoides biting midges: their role as arbovirus vectors. Annu Rev Entomol. 2000; 45: 307-40.
Harrup LE, Bellis GA, Balenghien T, Garros C.Culicoides Latreille (Diptera: Ceratopogonidae) taxonomy: current challenges and future directions. Infect Genet Evol. 2015; 30: 249-66.
Borkent A. The biting midges, the Ceratopogonidae (Diptera), biology of disease vectors, 2nd ed. Elsevier, Burlington, MA: USA; 2004: 113-126.
Meiswinkel R, Venter GJ, Nevill EM. Vectors:Culicoides spp. In: Coetzer JAW, Tustin RC, editors. Infectious Diseases of Livestock. Cape Town: Oxford University Press; 2004: 93-136.
Carpenter S, Groschup MH, Garros C, Felippe-Bauer MF, Purse B.Culicoides biting midges, arboviruses and public health in Europe. Antiviral Res. 2013; 13: 102-13.
OIE. OIE-Listed diseases, infections and infestations in force in 2014, updated: 1st January 2014, accessed: 17th January 2014. Available at: http://www.oie.int/animal-health-in-the-world/oie-listed-diseases-2014/.
Elbers AR, Meiswinkel R, van Weezep E, van Oldruitenborgh-Oosterbaan MM, Kooi EA. Schmallenberg virus inCulicoides spp. biting midges, the Netherlands, 2011. Emerg Infect Dis. 2013; 19: 106-9.
Valkiūnas G. Avian malaria parasites and other haemosporidia. Boca Raton: CRC Press; 2005.
Atkinson CT.Haemoproteus. In: Atkinson CT, Thomas NJ, Hunter BC, editors. Parasitic diseases of wild birds. Ames: Wiley-Blackwell; 2008. p.13-35.
Donovan TA, Schrenzel M, Tucker TA, Pessier AP, Stalis IH. Hepatic hemorrhage, hemocoelom, and sudden death due toHaemoproteus infection in passerine birds: eleven cases. J Vet Diagn Invest. 2008; 20: 304-13.
Ortiz-Catedral L, Brunton D, Stidworth MF, Elsheikha HM, Pennycott T, Schulze C, et al.Haemoproteus minutus is highly virulent for Australasian and South American parrots. Parasit Vector. 2019; 12: 40.
Borkent A. World species of biting midges (Diptera: Ceratopogonidae). New York: American Museum of Natural History; 2019.
Bukauskaitė D, Iezhova TA, Ilgūnas M, Valkiūnas G. High susceptibility of the laboratory-reared biting midgesCulicoides nubeculosus to Haemoproteus infections, with review on Culicoides species that transmit avian haemoproteids. Parasitology. 2019; 146: 1-9.
Bernotienė R, Žiegytė R, Vaitkutė G, Valkiūnas G. Identification of a new vector species of avian haemoproteids, with a description of methodology for the determination of natural vectors of haemosporidian parasites. Parasites & Vectors. 2019; 12: 307.
Iezhova TA, Dodge M, Sehgal RN, Smith TB, Valkiunas G. New avianHaemoproteus species (Haemosporida: Haemoproteidae) from African birds, with a critique of the use of host taxonomic information in hemoproteid classification. J Parasites. 2011; 97: 682-94.
Yıldırım A, Dik B, Düzlü Ö, Gürsoy Ergen A. Sultan sazlığı ekosisteminde kanatlı Haemosporidialarının moleküler ekolojisi. TÜBİTAK 1001 Projesi, Proje no: 114 O 646, 2017.
Kieffer JJ. Chironomides d’Afrique et d’Asie conservés au Museum National Hongrois de Budapest. Ann Hist Nat Mus Nat Hung. 1918; 16: 31-136.
Dik B.Culicoides (Diptera: Ceratopogonidae) vektörlükleri ve mücadelesi. İçinde: Özbel Y (editör), Vektör artropodlar ve mücadelesi. Türkiye Parazitoloji Derneği: İzmir; 2017. s.46-122.
Korkmaz C, Alten B, Erol U, Deniz A. Updated checklist ofCulicoides Latreille (Diptera: Ceratopogonidae) of Türkiye with ten new records. J Vector Ecol. 2021; 46: 124-36.
Turgut F. Morphological characteristics of twoCulicoides Latreille biting midges (Diptera, Ceratopogonidae) with a new record for Turkish fauna. Trakya Univ J Nat Sci. 2023; 23: 3-8.
Dik B, Muz D, Muz MN, Uslu U. The geographical distribution and first molecular analysis ofCulicoides Latreille (Diptera: Ceratopogonidae) species in the Southern and Southeastern Türkiye during the 2012 outbreak of bovine ephemeral fever. Parasitol Res. 2014; 113: 4225-32.
Yildirim A, Dik B, Duzlu O, Onder Z, Ciloglu A, Yetismis G, et al. Genetic diversity ofCulicoides species within the Pulicaris complex (Diptera: Ceratopogonidae) in Türkiye inferred from mitochondrial COI gene sequences. Acta Trop. 2019; 190: 380-8.
Şevik M, Oz ME. Detection of Peste des petits ruminants virus RNA inCulicoides imicola (Diptera: Ceratopogonidae) in Türkiye. Vet Ital. 2019; 55: 173-9.
Nolan DV, Carpenter S, Barber J, Mellor PS, Dallas JF, Mordue Luntz AJ, et al. Rapid diagnostic PCR assays for members of theCulicoides obsoletus and Culicoides pulicaris species complexes, implicated vectors of bluetongue virus in Europe. Vet Microbiol. 2007; 124: 82-94.
Pages N, Munoz-Munoz F, Talavera S, Sarto V, Lorca C, Núñez JI. Identification of cryptic species ofCulicoides (Diptera: Ceratopogonidae) in the subgenus Culicoides and development of species-specific PCR assays based on barcode regions. Vet Parasitol. 2009; 165: 298-310.
Mathieu B, Cêtre-Sossah C, Garros C, Chavernac D, Balenghien T, Carpenter S, et al. Development and validation of IIKC: an interactive identification key forCulicoides (Diptera: Ceratopogonidae) females from the Western Palaearctic region. Parasites & Vectors. 2012; 5: 137.
Dallas JF, Cruickshank RH, Linton YM, Nolan DV, Patakakis M, Braverman Y, et al. Phylogenetic status and matrilineal structure of the biting midge,Culicoides imicola, in Portugal, Rhodes and Israel. Med Vet Entomol. 2003; 17: 379-87.
Hellgren O, Waldenström J, Bensch S. A new PCR assay for simultaneous studies ofLeucocytozoon, Plasmodium, and Haemoproteus from avian blood. J Parasitol. 2004; 90: 797-802.
Bensch S, Hellgren O, Pérez-Tris J. MalAvi: A public database of malaria parasites and related haemosporidians in avian hosts based on mitochondrial cytochrome b lineages. Mol Ecol Resour. 2009; 9: 1353.
Kearse M, Moir R, Wilson A, Stones-Havas S, Cheung M, Sturrock S, et al. Geneious basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics. 2012; 28: 1647-9.
Librado P, Rozas J. DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics. 2009; 25: 1451-2.
Kimura M. A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. J Mol Evol. 1980; 16: 111-20.
Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Mol Biol Evol. 2013; 30: 2725-9.
Posada D. jModelTest: phylogenetic model averaging. Mol Biol Evol. 2008; 25: 1253-6.
Huelsenbeck JP, Ronquist F. MRBAYES: bayesian inference of phylogenetic trees. Bioinformatics. 2001; 17: 754-5.
Ramilo DW, Nunes T, Madeira S, Boinas F, da Fonseca IP. Geographical distribution ofCulicoides (Diptera: Ceratopogonidae) in mainland Portugal: presence/absence modelling of vector and potential vector species. PLoS One. 2017; 12: e0180606.
Sarvašová A, Kočišová A, Halán M, Delécolle JC, Mathieu B. Morphological and molecular analysis of the genusCulicoides (Diptera: Ceratopogonidae) in Slovakia with five new records. Zootaxa. 2014; 3872: 541-60.
Carvalho LPC, Costa GDS, Pereira Júnior AM, de Paulo PFM, Silva GS, Carioca ALPM, et al. DNA Barcoding of genusCulicoides biting midges (Diptera: Ceratopogonidae) in the Brazilian Amazon. Acta Trop. 2022; 235: 106619.
Gopurenko D, Bellis G, Pengsakul T, Siriyasatien P, Thepparat A. DNA barcoding ofCulicoides Latreille (Diptera: Ceratopogonidae) from Thailand reveals taxonomic inconsistencies and novel diversity among reported sequences. J Med Entomol. 2022; 59: 1960-70.
Ortega-Morales AI, Hernández-Triana LM, Chan-Chable RJ, Garza-Hernández JA, González-Álvarez VH, Ruiz-Arrondo I, et al. DNA barcoding of mosquitoes from the Pantanos de Centla biosphere reserve, Southeastern Mexico. J Am Mosq Control Assoc. 2021; 37: 198-207.
Dähn O, Werner D, Mathieu B, Kampen H. Large-scale cytochrome C Oxidase Subunit I gene data analysis for the development of a multiplex polymerase chain reaction test capable of identifying biting midge vector species and haplotypes (Diptera: ceratopogonidae) of theCulicoides Subgenus Avaritia Fox, 1955. Genes (Basel). 2024; 15: 323.
Onder Z, Yildirim A, Duzlu O, Arslan MO, Sari B, Tasci GT, et al. Molecular characterization of black flies (Diptera: Simuliidae) in areas with pest outbreaks and simuliotoxicosis in Northeast Anatolia Region, Türkiye. Acta Trop. 2019; 199: 105149.
Inci A, Yildirim A, Duzlu O, Onder Z, Ciloglu A, Seitz G, et al. Genetic diversity and identification of Palearctic black flies in the subgenus Wilhelmia (Diptera: Simuliidae). J Med Entomol. 2017; 54: 888-94.
Anon. İzmir Bornova Veteriner Kontrol Enstitüsü Kayıtları; 2016.
Goredo M, Catalani M, Federici V, Portanti O, Marini V, Mancini G, et al. Vector species ofCulicoides midges implicated in the 2012-2014 Bluetongue epidemics in Italy. Vet Ital. 2015; 51: 131-8.
Goredo M, Meiswinkel R, Federici V, Di Nicola F, Mancini G, Ippoliti C, et al. The “Culicoides obsoletus group” in Italy: Relative abundance, geographic range, and role as vector for Bluetongue virus. Vet Ital. 2016; 52: 235-24.
Yavru S, Dik B, Bulut O, Yavru N, Kale M, Bulut A. NewCulicoides vector species for BTV transmission in Central and Central west of Anatolia. Annu Res Rev Biol. 2018; 27: 1-9.
Foxi C, Meloni G, Puggioni G, Manunta D, Rocchigiani A, Vento L, et al. Bluetongue virus detection in newCulicoides species in Sardinia, Italy. Vet Re. 2019; 184: 621.
Dik B, Kuçlu Ö, Öztürk R.Culicoides Latreille, 1809 (Diptera: Ceratopogonidae) species in the Western Black Sea region of Türkiye, new records for the Turkish fauna. Turk J Vet Anim Sci. 2017; 41: 228-37.
Reyhan E. Güney Doğu Anadolu Bölgesinin Gaziantep, Kahramanmaraş ve Kilis illerindeCulicoides (Diptera: Ceratopogonidae) türlerinin yayılışı, Yüksek Lisans Tezi, Gaziantep Üniversitesi Fen Bilimleri Enstitüsü: Gaziantep; 2018.
Turgut F, Kilic AY. The Ceratopogonidae (Insecta: Diptera) fauna of the Central Black Sea region in Türkiye. Turk Zool Derg. 2015; 39: 1071-89.
Muz D, Dik B, Muz MN, Öztürk R. Tekirdağ iliCulicoides (Diptera: Ceratopogonidae) faunasının mavi dil virus ve Schmallenberg virus varlığı yönünden araştırılması. Namık Kemal Üniversitesi Araştırma Projesi: 2016, Proje no: NKUBAP.00.23.AR.14.08.
Turgut F. Samsun iliCulicoides Latreille, 1809 (Diptera: Ceratopogonidae) faunasına katkılar. GBAD. 2018; 7: 14-20.
Turgut F. Contributions toCulicoides Latreille, 1809 (Diptera: Ceratopogonidae) fauna of Sinop province. SAUJS. 2018; 22: 1683-93.
Hadj-Henni L, De Meulemeester T, Mathieu B, Depaquit J, Augot D. Taxonomic assessment ofCulicoides brunnicans, C. santonicus and C. vexans (Diptera: Ceratopogonidae) in France: Implications in systematics. Infect Genet Evol. 2015; 33: 324-31.
Augot D, Ninio C, Akhoundi M, Lehrter V, Couloux A, Jouet D, et al. Characterization of two cryptic species,Culicoides stigma and C. parroti (Diptera: Ceratopogonidae), based on barcode regions and morphology. J Vector Ecol. 2013; 38: 260-5.
Lassen SB, Nielsen SA, Skovgård H, Kristensen M. Molecular differentiation ofCulicoides biting midges (Diptera: Ceratopogonidae) from the subgenus Culicoides Latreille in Denmark. Parasitol Res. 2012; 110: 1765-71.
Pagès N, Sarto I Monteys V. Differentiation ofCulicoides obsoletus and Culicoides scoticus (Diptera: Ceratopogonidae) based on mitochondrial cytochrome oxidase subunit I. J Med Entomol. 2005; 42: 1026-34.
Ishtiaq F, Guillaumot L, Clegg SM, Phillimore AB, Black RA, Owens IP, et al. Avian haematozoan parasites and their associations with mosquitoes across Southwest Pacific Islands. Mol Ecol. 2008; 17: 4545-55.
Njabo KY, Cornel AJ, Bonneaud C, Toffelmier E, Sehgal RN, Valkiūnas G, et al. Nonspecific patterns of vector, host and avian malaria parasite associations in a central African rainforest. Mol Ecol. 2011; 20: 1049-61.
James AA, Rossignol PA. Mosquito salivary glands: parasitological and molecular aspects. Parasitol Today. 1991; 7: 267-71.
Inci A, Yıldırım A, Njabo KY, Duzlu O, Biskin Z, Ciloglu A. Detection and molecular characterization of avianPlasmodium from mosquitoes in central Türkiye. Vet Parasitol. 2012; 188: 179-84.
Mata VA, da Silva LP, Lopes RJ, Drovetski SV. The strait of gibraltar poses an effective barrier to host-specialised but not to host-generalised lineages of avian Haemosporidia. Int J Parasitol. 2015; 45: 711-9.
Martinez-de la Puente J, Munoz J, Capelli G, Montarsi F, Soriguer R, Arnoldi D, et al. Avian malaria parasites in the last supper: identifying encounters between parasites and the invasive Asian mosquito tiger and native mosquito species in Italy. Malaria J. 2015; 14: 32.
Synek P, Munclinger P, Albrecht T, Votypka J. Avian haemosporidians in haematophagous insects in the Czech Republic. Parasitol Res. 2013; 112: 839-45.
Bobeva A, Zehtindjiev P, Bensch S, Radrova J. A survey of biting midges of the genusCulicoides Latreille, 1809 (Diptera: Ceratopogonidae) in NE Bulgaria, with respect to transmission of avian haemosporidians. Acta Parasitol. 2013; 58: 585-91.
Ferraguti M, Martinez-de la Puente J, Munoz J, Roiz D, Ruiz S, Soriguer R, et al. AvianPlasmodium in Culex and Ochlerotatus mosquitoes from Southern Spain: effects of season and host-feeding source on parasite dynamics. PLoS One. 2013; 8: e66237.
Kim KS, Tsuda Y. AvianPlasmodium lineages found in spot surveys of mosquitoes from 2007 to 2010 at Sakata wetland, Japan: do dominant lineages persist for multiple years? Mol Ecol. 2012; 21: 5374-85.
