An integrative taxonomic approach reveals two putatively novel species of phlebotomine sand fly (Diptera: Psychodidae) in Thailand

Abstract Background The subfamily Phlebotominae comprises 1028 species of sand fly, of which only 90 are recognized as vectors of pathogenic agents such as Trypanosoma, Leishmania, and Bartonella. In Thailand, leishmaniasis—a sand fly-borne disease—is currently endemic, with 36 documented sand fly s...

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Main Authors: Samiullah Soomro, Siwaporn Tuangpermsub, Thongchai Ngamprasertwong, Morakot Kaewthamasorn
Format: Article
Language:English
Published: BMC 2025-01-01
Series:Parasites & Vectors
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Online Access:https://doi.org/10.1186/s13071-024-06640-8
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author Samiullah Soomro
Siwaporn Tuangpermsub
Thongchai Ngamprasertwong
Morakot Kaewthamasorn
author_facet Samiullah Soomro
Siwaporn Tuangpermsub
Thongchai Ngamprasertwong
Morakot Kaewthamasorn
author_sort Samiullah Soomro
collection DOAJ
description Abstract Background The subfamily Phlebotominae comprises 1028 species of sand fly, of which only 90 are recognized as vectors of pathogenic agents such as Trypanosoma, Leishmania, and Bartonella. In Thailand, leishmaniasis—a sand fly-borne disease—is currently endemic, with 36 documented sand fly species. However, many cryptic species likely remain unidentified. To improve our understanding of the distribution, habitat preferences, and role in disease transmission of these sand flies, further research is necessary. Methods Sand flies were collected using CDC light traps from 13 locations across four provinces in Thailand between October 2022 and October 2023. Initially, species identification was based on morphological characteristics, employing identification keys, and subsequently confirmed through mitochondrial cytochrome oxidase c subunit I (COI) and cytochrome b (Cytb) sequencing. Species identities were verified using BLASTN and BOLD searches. Species delimitation was conducted using Automatic Barcode Gap Discovery (ABGD) and Assemble Species by Automatic Partitioning (ASAP) with three substitution models. Additionally, intraspecific and interspecific genetic variation, neutrality tests (including Tajima’s and Fu and Li’s D* tests), phylogenetic analyses, and TCS haplotype network analysis were performed using the obtained sequences. Results A total of 3693 phlebotomine sand flies were collected, with 2261 (61.22%) identified as female. Integrative analyses combining morphological data, BLASTN searches, phylogenetic assessments, and species delimitation confirmed the identification of four genera: Sergentomyia, Grassomyia, Phlebotomus, and Idiophlebotomus, encompassing 12 species: Sergentomyia anodontis, Se. sylvatica, Se. perturbans, Se. barraudi, Se. hivernus, Se. khawi, Se. siamensis, Grassomyia indica, Phlebotomus barguesae, Ph. stantoni, Idiophlebotomus asperulus, and Id. longiforceps. Furthermore, molecular analysis revealed cryptic and complex species, including two putatively novel species, Se. sp. 1 and Se. sp. 2, as well as a unique haplotype. Conclusions This study, which integrated genetic and morphological identification techniques, identified 12 sand fly species and unveiled cryptic and complex species, including two putatively novel species (Se. sp. 1 and Se. sp. 2) and a unique haplotype. The findings underscore the utility of mitochondrial genes, combined with species delimitation methodologies, as reliable approaches for identifying diverse sand fly species. Graphical Abstract
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spelling doaj-art-54e00a7f3a74470eb42aa28d40ad547c2025-01-12T12:09:58ZengBMCParasites & Vectors1756-33052025-01-0118112110.1186/s13071-024-06640-8An integrative taxonomic approach reveals two putatively novel species of phlebotomine sand fly (Diptera: Psychodidae) in ThailandSamiullah Soomro0Siwaporn Tuangpermsub1Thongchai Ngamprasertwong2Morakot Kaewthamasorn3The International Graduate Program of Veterinary Science and Technology (VST), Faculty of Veterinary Science, Chulalongkorn UniversityCenter of Excellence in Veterinary Parasitology, Department of Pathology, Faculty of Veterinary Science, Chulalongkorn UniversityDepartment of Biology, Faculty of Science, Chulalongkorn UniversityCenter of Excellence in Veterinary Parasitology, Department of Pathology, Faculty of Veterinary Science, Chulalongkorn UniversityAbstract Background The subfamily Phlebotominae comprises 1028 species of sand fly, of which only 90 are recognized as vectors of pathogenic agents such as Trypanosoma, Leishmania, and Bartonella. In Thailand, leishmaniasis—a sand fly-borne disease—is currently endemic, with 36 documented sand fly species. However, many cryptic species likely remain unidentified. To improve our understanding of the distribution, habitat preferences, and role in disease transmission of these sand flies, further research is necessary. Methods Sand flies were collected using CDC light traps from 13 locations across four provinces in Thailand between October 2022 and October 2023. Initially, species identification was based on morphological characteristics, employing identification keys, and subsequently confirmed through mitochondrial cytochrome oxidase c subunit I (COI) and cytochrome b (Cytb) sequencing. Species identities were verified using BLASTN and BOLD searches. Species delimitation was conducted using Automatic Barcode Gap Discovery (ABGD) and Assemble Species by Automatic Partitioning (ASAP) with three substitution models. Additionally, intraspecific and interspecific genetic variation, neutrality tests (including Tajima’s and Fu and Li’s D* tests), phylogenetic analyses, and TCS haplotype network analysis were performed using the obtained sequences. Results A total of 3693 phlebotomine sand flies were collected, with 2261 (61.22%) identified as female. Integrative analyses combining morphological data, BLASTN searches, phylogenetic assessments, and species delimitation confirmed the identification of four genera: Sergentomyia, Grassomyia, Phlebotomus, and Idiophlebotomus, encompassing 12 species: Sergentomyia anodontis, Se. sylvatica, Se. perturbans, Se. barraudi, Se. hivernus, Se. khawi, Se. siamensis, Grassomyia indica, Phlebotomus barguesae, Ph. stantoni, Idiophlebotomus asperulus, and Id. longiforceps. Furthermore, molecular analysis revealed cryptic and complex species, including two putatively novel species, Se. sp. 1 and Se. sp. 2, as well as a unique haplotype. Conclusions This study, which integrated genetic and morphological identification techniques, identified 12 sand fly species and unveiled cryptic and complex species, including two putatively novel species (Se. sp. 1 and Se. sp. 2) and a unique haplotype. The findings underscore the utility of mitochondrial genes, combined with species delimitation methodologies, as reliable approaches for identifying diverse sand fly species. Graphical Abstracthttps://doi.org/10.1186/s13071-024-06640-8Genetic diversityPhlebotominePhylogeneticsSpecies delimitationTCS haplotype networkThailand
spellingShingle Samiullah Soomro
Siwaporn Tuangpermsub
Thongchai Ngamprasertwong
Morakot Kaewthamasorn
An integrative taxonomic approach reveals two putatively novel species of phlebotomine sand fly (Diptera: Psychodidae) in Thailand
Parasites & Vectors
Genetic diversity
Phlebotomine
Phylogenetics
Species delimitation
TCS haplotype network
Thailand
title An integrative taxonomic approach reveals two putatively novel species of phlebotomine sand fly (Diptera: Psychodidae) in Thailand
title_full An integrative taxonomic approach reveals two putatively novel species of phlebotomine sand fly (Diptera: Psychodidae) in Thailand
title_fullStr An integrative taxonomic approach reveals two putatively novel species of phlebotomine sand fly (Diptera: Psychodidae) in Thailand
title_full_unstemmed An integrative taxonomic approach reveals two putatively novel species of phlebotomine sand fly (Diptera: Psychodidae) in Thailand
title_short An integrative taxonomic approach reveals two putatively novel species of phlebotomine sand fly (Diptera: Psychodidae) in Thailand
title_sort integrative taxonomic approach reveals two putatively novel species of phlebotomine sand fly diptera psychodidae in thailand
topic Genetic diversity
Phlebotomine
Phylogenetics
Species delimitation
TCS haplotype network
Thailand
url https://doi.org/10.1186/s13071-024-06640-8
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