Discrimination of Phlebotomus perfiliewi transcaucasicus, Ph. major sensu lato and Ph. tobbi (Diptera: Psychodidae) Using Morphometric and DNA Barcoding Methods in the Endemic Foci of Visceral Leishmaniasis in Ardabil Province, North West of Iran

Background: Visceral leishmaniasis, commonly known as kala-azar, and prevalent in more than 70 countries and sev­eral regions of Iran. It is one of the main diseases transmitted by sand flies. In this work, geometric morphometrics and DNA barcoding were employed as novel techniques to enhance the d...

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Main Authors: Alireza Barmaki, Yavar Rassi, Azad Absavaran, Amir Ahmad Akhavan, Eslam Moradi-Asl, Alireza Zahraei-Ramazani, Sayena Rafizadeh
Format: Article
Language:English
Published: Tehran University of Medical Sciences 2025-03-01
Series:Journal of Arthropod-Borne Diseases
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Online Access:https://jad.tums.ac.ir/index.php/jad/article/view/1704
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author Alireza Barmaki
Yavar Rassi
Azad Absavaran
Amir Ahmad Akhavan
Eslam Moradi-Asl
Alireza Zahraei-Ramazani
Sayena Rafizadeh
author_facet Alireza Barmaki
Yavar Rassi
Azad Absavaran
Amir Ahmad Akhavan
Eslam Moradi-Asl
Alireza Zahraei-Ramazani
Sayena Rafizadeh
author_sort Alireza Barmaki
collection DOAJ
description Background: Visceral leishmaniasis, commonly known as kala-azar, and prevalent in more than 70 countries and sev­eral regions of Iran. It is one of the main diseases transmitted by sand flies. In this work, geometric morphometrics and DNA barcoding were employed as novel techniques to enhance the diagnos­tic tools used in this study. Methods: Phlebotomus perfiliewi transcaucasicus, Phlebotomus major s.l., and Phlebotomus tobbi caught from three districts in the Ardabil Province, northwest of Iran. The right wings of 286 female sand flies were analyzed using geo­metric morphometric (GM) tools. Additionally, the COI gene was isolated from each of the three species, amplified using universal primers, and sequenced through the DNA barcoding method for classification. This sequencing data was then formatted to generate morphometric analyses. Results: The landmarks with the most variations were found in sets 10, 12, 13, and 14, whereas the first set's landmarks at 1 and 11, along with those from the second set at positions 2, 3, and 5 exhibited the greatest variations. Analysis of the size and shape variations in the wings indicates the presence of distinct populations (P< 0.05). Furthermore, the DNA barcoding results not only confirmed the findings from the geometric morphometric analysis but also revealed both interspecific and intraspecific distances. Conclusion: This study was the first attempt to assess whether wing geometry morphometrics, combined with DNA barcode techniques, can effectively distinguish the three mentioned species in the studied areas. Furthermore, the identi­fication of Phlebotomus neglectus in this area prompted recommendations for additional research.
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record_format Article
series Journal of Arthropod-Borne Diseases
spelling doaj-art-99cbddced9d64df2aa08758939ca80232025-08-20T03:26:39ZengTehran University of Medical SciencesJournal of Arthropod-Borne Diseases2322-19842322-22712025-03-0118310.18502/jad.v18i3.18573Discrimination of Phlebotomus perfiliewi transcaucasicus, Ph. major sensu lato and Ph. tobbi (Diptera: Psychodidae) Using Morphometric and DNA Barcoding Methods in the Endemic Foci of Visceral Leishmaniasis in Ardabil Province, North West of IranAlireza Barmaki0Yavar Rassi1Azad Absavaran2Amir Ahmad Akhavan3Eslam Moradi-Asl4Alireza Zahraei-Ramazani5Sayena Rafizadeh6Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, Department of Public Health, School of Public Health, Qazvin University of Medical Sciences, Qazvin, IranDepartment of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran, Arthropod-Borne Diseases Research Center, Ardabil University of Medical Sciences, Ardabil, IranDepartment of Vector Biology and Control of Diseases, School of Medicine, Zanjan University of Medical Sciences, Zanjan, IranDepartment of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran3Arthropod-Borne Diseases Research Center, Ardabil University of Medical Sciences, Ardabil, Iran, Department of Public Health, School of Public Health, Ardabil University of Medical Sciences, Ardabil, IranDepartment of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, IranMinistry of Health and Medical Education, Tehran, Iran Background: Visceral leishmaniasis, commonly known as kala-azar, and prevalent in more than 70 countries and sev­eral regions of Iran. It is one of the main diseases transmitted by sand flies. In this work, geometric morphometrics and DNA barcoding were employed as novel techniques to enhance the diagnos­tic tools used in this study. Methods: Phlebotomus perfiliewi transcaucasicus, Phlebotomus major s.l., and Phlebotomus tobbi caught from three districts in the Ardabil Province, northwest of Iran. The right wings of 286 female sand flies were analyzed using geo­metric morphometric (GM) tools. Additionally, the COI gene was isolated from each of the three species, amplified using universal primers, and sequenced through the DNA barcoding method for classification. This sequencing data was then formatted to generate morphometric analyses. Results: The landmarks with the most variations were found in sets 10, 12, 13, and 14, whereas the first set's landmarks at 1 and 11, along with those from the second set at positions 2, 3, and 5 exhibited the greatest variations. Analysis of the size and shape variations in the wings indicates the presence of distinct populations (P< 0.05). Furthermore, the DNA barcoding results not only confirmed the findings from the geometric morphometric analysis but also revealed both interspecific and intraspecific distances. Conclusion: This study was the first attempt to assess whether wing geometry morphometrics, combined with DNA barcode techniques, can effectively distinguish the three mentioned species in the studied areas. Furthermore, the identi­fication of Phlebotomus neglectus in this area prompted recommendations for additional research. https://jad.tums.ac.ir/index.php/jad/article/view/1704Visceral leishmaniasisPhlebotomus perfiliewi transcaucasicusDNA barcodingIran
spellingShingle Alireza Barmaki
Yavar Rassi
Azad Absavaran
Amir Ahmad Akhavan
Eslam Moradi-Asl
Alireza Zahraei-Ramazani
Sayena Rafizadeh
Discrimination of Phlebotomus perfiliewi transcaucasicus, Ph. major sensu lato and Ph. tobbi (Diptera: Psychodidae) Using Morphometric and DNA Barcoding Methods in the Endemic Foci of Visceral Leishmaniasis in Ardabil Province, North West of Iran
Journal of Arthropod-Borne Diseases
Visceral leishmaniasis
Phlebotomus perfiliewi transcaucasicus
DNA barcoding
Iran
title Discrimination of Phlebotomus perfiliewi transcaucasicus, Ph. major sensu lato and Ph. tobbi (Diptera: Psychodidae) Using Morphometric and DNA Barcoding Methods in the Endemic Foci of Visceral Leishmaniasis in Ardabil Province, North West of Iran
title_full Discrimination of Phlebotomus perfiliewi transcaucasicus, Ph. major sensu lato and Ph. tobbi (Diptera: Psychodidae) Using Morphometric and DNA Barcoding Methods in the Endemic Foci of Visceral Leishmaniasis in Ardabil Province, North West of Iran
title_fullStr Discrimination of Phlebotomus perfiliewi transcaucasicus, Ph. major sensu lato and Ph. tobbi (Diptera: Psychodidae) Using Morphometric and DNA Barcoding Methods in the Endemic Foci of Visceral Leishmaniasis in Ardabil Province, North West of Iran
title_full_unstemmed Discrimination of Phlebotomus perfiliewi transcaucasicus, Ph. major sensu lato and Ph. tobbi (Diptera: Psychodidae) Using Morphometric and DNA Barcoding Methods in the Endemic Foci of Visceral Leishmaniasis in Ardabil Province, North West of Iran
title_short Discrimination of Phlebotomus perfiliewi transcaucasicus, Ph. major sensu lato and Ph. tobbi (Diptera: Psychodidae) Using Morphometric and DNA Barcoding Methods in the Endemic Foci of Visceral Leishmaniasis in Ardabil Province, North West of Iran
title_sort discrimination of phlebotomus perfiliewi transcaucasicus ph major sensu lato and ph tobbi diptera psychodidae using morphometric and dna barcoding methods in the endemic foci of visceral leishmaniasis in ardabil province north west of iran
topic Visceral leishmaniasis
Phlebotomus perfiliewi transcaucasicus
DNA barcoding
Iran
url https://jad.tums.ac.ir/index.php/jad/article/view/1704
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