Phylogenetic characterization of two common sandflies, Phlebotomus major and P. kandelakii, in Inebolu District of the West Black Sea Region, Türkiye based on mitochondrial gene sequence analysis

Year 2024, Accepted Papers, 1 - 10

Gupse Kübra Karademir Mübeccel Atelge Kardelen Yetişmiş Gamze Yetişmiş Sadullah Uslu Arif Çiloğlu Zuhal Önder Yusuf Özbel Gökmen Zafer Pekmezci Alparslan Yıldırım Önder Düzlü Seray Töz Didem Pekmezci Abdullah İnci

https://doi.org/10.33988/auvfd.1372324

Abstract

Research on the taxonomy and phylogenetic characterization of sand flies (Diptera, Psychodidae, Phlebotominae) has received increasing attention due to their potential role in the transmission of medically significant diseases, including leishmaniasis, bartonellosis, and sand fly fever. Phlebotomus major and Phlebotomus kandelakii are the members of Larroussius subgenus that includes important vector sand fly species. P. major, P. neglectus and P. syriacus from a well-known sand fly “Major” species group are present widely while P. kandelakii has constricted distribution in Turkey. All have the capability to transmit Leishmania infantum, the causative agent of visceral leishmaniasis. Here, we investigated the genetic diversity within each species collected from the West Black Sea Region of Turkey using mitochondrial DNA markers, specifically cytochrome oxidase I (COI) and cytochrome b gene sequences (Cytb). A total of 1889 sand flies specimens were collected from the study area in June 2021 and August 2022, 1596 (84.49%) were identified as P. major s.l., and 253 (13.40%) were identified as P. kandelakii. Nine and four haplotypes of P. major were determined in the study area based on COI and Cytb sequences, respectively. Analysis of the phylogenetic data sets generated from our isolates and published isolates in Genbank revealed high haplotype diversities within P. major (COI = 0.933, Cytb = 0.714). For P. kandelakii, we detected four and three haplotypes within the COI and Cytb sequences, and the haplotype diversities were also high in the datasets including our isolates and published isolates in Genbank (COI = 0.978, Cytb = 1.000). Pairwise mean genetic distance calculated from the COI and Cytb datasets were 0.4% and 1.4% for P. major, and 1.0% and 0.2% for P. kandelakii, respectively, suggesting the absence of cryptic species. Phylogenetic analyses revealed three and two major clusters of Larroussius subgenus in the COI and Cytb datasets, respectively.

Keywords

COI, Cytb, haplotype, Sand fly, West Black Sea Region; Turkey

Supporting Institution

Erciyes University Scientific Research Project Coordination Unit

Project Number

TCD-2021-11185

Thanks

Gupse Kubra Karademir and Sadullah Uslu were supported by the Council of Higher Education (YOK) 100/2000 PhD scholarship. Thanks to YOK for 100/2000 PhD scholarship.

References

• Adler S, Theodor O (1931): Investigations on mediterranean kala azar. V.-Distribution of sandflies of the major group in relation to Mediterranean kala azar. Proceedings of the Royal Society of London Series B, Containing Papers of a Biological Character, 108, 494–502.
• Artem’ev MM, Neronov VM (1984): Distribution and ecology of sandflies of the Old World (genus Phlebotomus). Moscow: Institut Ėvolyutsionnoĭ Morfologii Ėkologii Zhivotnykh, 207.
• Badakhshan M, Sadraei J, Moin‐Vaziri V (2011): Morphometric and morphological variation between two different populations of Phlebotomus major sl from endemic and non‐endemic foci of visceral leishmaniasis in Iran. Journal of Vector Ecology, 36, 153–158.
• Bailey F, Mondragon-Shem K, Hotez P, et al (2017): A new perspective on cutaneous leishmaniasis—Implications for global prevalence and burden of disease estimates. PLoS Neglected Tropical Diseases, 11, e0005739.
• Bates PA, Depaquit J, Galati EA, et al (2015): Recent advances in phlebotomine sand fly research related to leishmaniasis control. Parasites & Vectors, 8, 1–8.
• Brilhante AF, de Albuquerque AL, Rocha AC de B, et al (2020): First report of an Onchocercidae worm infecting Psychodopygus carrerai carrerai sandfly, a putative vector of Leishmania braziliensis in the Amazon. Scientific Reports, 10, 15246.
• Chen H, Dong H, Yuan H, et al (2023): Mitochondrial COI and Cytb gene as valid molecular identification marker of sandfly species (Diptera: Psychodidae) in China. Acta Tropica, 238, 106798.
• Contreras Gutierrez MA, Vivero RJ, Velez ID, et al (2014): DNA barcoding for the identification of sand fly species (Diptera, Psychodidae, Phlebotominae) in Colombia. PloS One, 9, e85496.
• Depaquit J (2014): Molecular systematics applied to Phlebotomine sandflies: Review and perspectives. Infection, Genetics and Evolution, 28, 744–756.
• Depaquit J, Grandadam M, Fouque F, et al (2010): Arthropod-borne viruses transmitted by Phlebotomine sandflies in Europe: a review. Eurosurveillance, 15, 19507.
• Dokianakis E, Tsirigotakis N, Christodoulou V, et al (2018): Identification of wild-caught phlebotomine sand flies from Crete and Cyprus using DNA barcoding. Parasites & Vectors, 11, 1–9.
• El Kacem S, Ait Kbaich M, Mhaidi I, et al (2023): Population Genetic Structure of Phlebotomus sergenti (Diptera: Psychodidae) Collected in Four Regions of Morocco Based on the Analysis of Cyt b and EF-1α Genes. Journal of Medical Entomology, 60, 294–305.
• Erisoz Kasap O, Linton Y-M, Karakus M, et al (2019): Revision of the species composition and distribution of Turkish sand flies using DNA barcodes. Parasites Vectors, 12, 1–20.
• Fayaz S, Raz A, Bahrami F, et al (2023): Molecular identification of Phlebotomus kandelakii apyrase and assessment of the immunogenicity of its recombinant protein in BALB/c mice. Scientific Reports, 13, 8766.
• Florin DA, Rebollar-Téllez EA (2013): Divergence of Lutzomyia (Psathyromyia) shannoni (Diptera: Psychodidae: Phlebotominae) is indicated by morphometric and molecular analyses when examined between taxa from the southeastern United States and southern Mexico. Journal of Medical Entomology, 50, 1324–1329.
• Fu Y-X (1997): Statistical tests of neutrality of mutations against population growth, hitchhiking and background selection. Genetics, 147, 915–925.
• Guindon S, Gascuel O (2003): A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Systematic Biology, 52, 696–704.
• Gutierrez MAC, Lopez ROH, Ramos AT, et al (2021): DNA barcoding of Lutzomyia longipalpis species complex (Diptera: Psychodidae), suggests the existence of 8 candidate species. Acta Tropica, 221, 105983.
• Hebert PD, Ratnasingham S, De Waard JR (2003): Barcoding animal life: cytochrome c oxidase subunit 1 divergences among closely related species. Proceedings of the Royal Society of London Series B: Biological Sciences, 270, S96–S99.
• Huelsenbeck JP, Ronquist F (2001): MRBAYES: Bayesian inference of phylogenetic trees. Bioinformatics, 17, 754–755.
• Kasap OE, Votýpka J, Alten B (2013): The distribution of the Phlebotomus major complex (Diptera: Psychodidae) in Turkey. Acta Tropica, 127, 204–211.
• Katoh K, Standley DM (2013): MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Molecular Biology and Evolution, 30, 772–780.
• Keser EM (2013): İnebolu İlçe Analizi. KUZKA (Kuzey Anadolu Kalkınma Ajansı), Planlama, Programlama ve Stratejik Araştırmalar Birimi, 18, 20–38.
• Killick-Kendrick R, Tang Y, Killick-Kendrick M, et al (1991): The identification of female sandflies of the subgenus Larroussius by the morphology of the spermathecal ducts. Parassitologia, 33, 335–347.
• Kimura M (1980): A simple method for estimating evolutionary rates of base substitutions through comparative studies of nucleotide sequences. Journal of Molecular Evolution, 16, 111–120.
• Krüger A, Strüven L, Post RJ, et al (2011): The sandflies (Diptera: Psychodidae, Phlebotominae) in military camps in northern Afghanistan (2007–2009), as identified by morphology and DNA ‘barcoding’. Annals of Tropical Medicine & Parasitology, 105, 163–176.
• Kumar NP, Srinivasan R, Jambulingam P (2012): DNA barcoding for identification of sand flies (Diptera: Psychodidae) in India. Molecular Ecology Resources, 12, 414–420.
• Léger N, Pesson B (1987): Sur la taxonomie et la répartition géographique de Phlebotomus (Adlerius) Chinensis sl et P. Larroussius major sl (Psychodidae-Diptera): statut des espèces présentes en Grèce. Bulletin de La Société de Pathologie Exotique, 80, 252–260.
• Lewis DJ (1982): A taxonomic review of the genus Phlebotomus (Diptera: Psychodidae). Bulletin of the British Museum (Natural History), Entomology Series 52, 1–35.
• Librado P, Rozas J (2009): DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics, 25, 1451–1452.
• Lozano-Sardaneta YN, Paternina LE, Sanchez-Montes S, et al (2020): DNA barcoding and fauna of phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) from Los Tuxtlas, Veracruz, Mexico. Acta Tropica, 201, 105220.
• Maroli M, Feliciangeli MD, Bichaud L, et al (2013): Phlebotomine sandflies and the spreading of leishmaniases and other diseases of public health concern. Medical and Veterinary Entomology, 27, 123–147.
• Mozaffari E, Vatandoost H, Rassi Y, et al (2020): Epidemiology of visceral leishmaniasis with emphasis on the dynamic activity of sand flies in an important endemic focus of disease in Northwestern Iran. Journal of Arthropod-Borne Diseases, 14, 97.
• Nzelu CO, Cáceres AG, Arrunátegui-Jiménez MJ, et al (2015): DNA barcoding for identification of sand fly species (Diptera: Psychodidae) from leishmaniasis-endemic areas of Peru. Acta Tropica, 145, 45–51.
• Oryan A, Akbari M (2016): Worldwide risk factors in leishmaniasis. Asian Pacific Journal of Tropical Medicine, 9, 925–932.
• Ozbel Y, Toz S, Kitapcıoglu G (2019): Sark Cıbanı. Vol. 1. 1 ed. İzmir, Meta Basım.
• Parvizi P, Amirkhani A (2008): Mitochondrial DNA characterization of Sergentomyia sintoni populations and finding mammalian Leishmania infections in this sandfly by using ITS-rDNA gene. Iranian Journal of Vet Research, 9-18.
• Pavlou C, Dokianakis E, Tsirigotakis N, et al (2022): A molecular phylogeny and phylogeography of Greek Aegean Island sand flies of the genus Phlebotomus (Diptera: Psychodidae). Arthropod Systematics & Phylogeny, 80, 137–154.
• Perfiliev PP (1966): Sandflies (Family Phlebotomidae). 93, 382. In: O Theodor (Ed): Fauna SSSR Wiener Bindery Ltd, Jerusalem.
• Perrotey S (1998): Etude critique des caracteres et de leurs etats utilises pour la diagnose des plebotomes femelles (diptera: psychodidae) (doctorat: parasitologie). Reims.
• Pinto I de S, Chagas BD das, Rodrigues AAF, et al (2015): DNA barcoding of neotropical sand flies (Diptera, Psychodidae, Phlebotominae): species identification and discovery within Brazil. PLoS One, 10, e0140636.
• Posada D (2008): jModelTest: phylogenetic model averaging. Molecular Biology and Evolution, 25, 1253–1256.
• Ramírez-Soriano A, Ramos-Onsins SE, Rozas J, et al (2008): Statistical Power Analysis of Neutrality Tests Under Demographic Expansions, Contractions and Bottlenecks With Recombination. Genetics, 179, 555–567.
• Ramos-Onsins SE, Rozas J (2002): Statistical Properties of New Neutrality Tests Against Population Growth. Molecular Biology and Evolution, 19, 2092–2100.
• Rassi Y, Abai MR, Oshaghi MA, et al (2012): First detection of Leishmania infantum in Phlebotomus kandelakii using molecular methods in north-eastern Islamic Republic of Iran. EMHJ-Eastern Mediterranean Health Journal, 18, 387-392.
• Schwarz G (1978): Estimating thedimension of a model. Annals of Statistics, 6, 461–464.
• Seccombe AK, Ready PD, Huddleston LM (1993): A Catalogue of Old World phlebotomine sandflies (Diptera: Psychodidae, Phlebotominae). Occasional Papers on Systematic Entomology, 8, 1–57.
• Tajima F (1989): Statistical method for testing the neutral mutation hypothesis by DNA polymorphism. Genetics, 123, 585–595.
• Tamura K, Stecher G, Kumar S (2021): MEGA11: Molecular Evolutionary Genetics Analysis Version 11. Molecular Biology and Evolution, 38, 3022–3027.