First record of mermithid parasitism in adult biting midges, Culicoides huffi (Diptera: Ceratopogonidae), collected from Southern Thailand, with ultrastructural and molecular characterization

Abstract Background Culicoides biting midges are known vectors of several pathogens, including arboviruses, protozoa, and filarial worms. Additionally, mermithid nematodes have been found to parasitize Culicoides midges, causing pathogenic effects that reduce host fitness and often lead to death. Co...

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Main Authors: Chulaluk Promrangsee, Vivornpun Sanprasert, Arunrat Thepparat, Sakone Sunantaraporn, Rinnara Ampol, Rungfar Boonserm, Padet Siriyasatien, Kanok Preativatanyou
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Parasites & Vectors
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Online Access:https://doi.org/10.1186/s13071-025-06958-x
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author Chulaluk Promrangsee
Vivornpun Sanprasert
Arunrat Thepparat
Sakone Sunantaraporn
Rinnara Ampol
Rungfar Boonserm
Padet Siriyasatien
Kanok Preativatanyou
author_facet Chulaluk Promrangsee
Vivornpun Sanprasert
Arunrat Thepparat
Sakone Sunantaraporn
Rinnara Ampol
Rungfar Boonserm
Padet Siriyasatien
Kanok Preativatanyou
author_sort Chulaluk Promrangsee
collection DOAJ
description Abstract Background Culicoides biting midges are known vectors of several pathogens, including arboviruses, protozoa, and filarial worms. Additionally, mermithid nematodes have been found to parasitize Culicoides midges, causing pathogenic effects that reduce host fitness and often lead to death. Consequently, mermithids have potential as biocontrol agents to reduce midge populations. However, the biology of these entomopathogenic nematodes infecting Culicoides in Thailand remains unknown. Methods As part of the leishmaniasis surveillance program in Nakhon Si Thammarat Province, Southern Thailand, we collected Culicoides midges near the residence of a leishmaniasis patient in November 2024. The Culicoides samples were morphologically identified and examined microscopically for parasitic nematodes. Infected midges were dissected to isolate nematodes from each individual. The nematodes were then characterized morphologically using scanning electron microscopy (SEM) and identified molecularly via polymerase chain reaction (PCR) targeting the non-filarial hypervariable region I of the small subunit ribosomal RNA gene (SSU HVR-I) and mermithid small subunit ribosomal RNA (SSU rRNA) gene, followed by nanopore sequencing, phylogenetic analysis, and species delimitation testing. Results A total of 263 field-caught adult Culicoides midges were collected, with Culicoides huffi of the Calvipalpis group being the most abundant species (n = 155, 58.9%). Among these, 35 C. huffi samples, including 4 males, 11 females, and 20 intersex males, were infected with nematodes, resulting in an overall infection rate of 13.3%. The parasitized intersex males, presumably genetically male, exhibited a high degree of feminization in their antennae and wings, which likely enhances female-like behaviors such as detecting and flying toward breeding sites, possibly facilitating parasite transmission and reproductive success. The SEM analysis revealed key morphological features consistent with parasitic nematode larvae of the Mermithidae family, including a long, slender body, a stylet, cephalic papillae, amphids, and a trophosome. Basic Local Alignment Search Tool for nucleotide (BLASTn) analysis of non-filarial SSU HVR-I and mermithid-specific SSU rRNA gene sequences identified all nematodes as mermithids, showing 94.2–94.4% similarity to Pheromermis sp. from the hornet and 97.3% similarity to Mermis sp. from Culicoides obsoletus. Phylogenetic analysis and species delimitation suggest that these sequences represent a single putative species distinct from other known mermithids. Conclusions This study is the first to report mermithid parasitism in Culicoides midges in Thailand, incorporating both ultrastructural and molecular characterization. The novel morphological and molecular insights provided here contribute to a more comprehensive understanding of the biology of entomopathogenic nematodes. Further research is needed to evaluate the potential of these nematodes for the biological control of Culicoides biting midges. Graphical Abstract
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spelling doaj-art-cd81c8154c704a0aa79b2a801f4bed912025-08-20T04:01:52ZengBMCParasites & Vectors1756-33052025-07-0118111510.1186/s13071-025-06958-xFirst record of mermithid parasitism in adult biting midges, Culicoides huffi (Diptera: Ceratopogonidae), collected from Southern Thailand, with ultrastructural and molecular characterizationChulaluk Promrangsee0Vivornpun Sanprasert1Arunrat Thepparat2Sakone Sunantaraporn3Rinnara Ampol4Rungfar Boonserm5Padet Siriyasatien6Kanok Preativatanyou7Interdisciplinary Program of Biomedical Sciences, Graduate School, Chulalongkorn UniversityDepartment of Parasitology, Faculty of Medicine, Chulalongkorn UniversityDepartment of Agricultural Technology, Faculty of Science, Ramkhamhaeng UniversityCenter of Excellence in Vector Biology and Vector-Borne Disease, Chulalongkorn UniversityDepartment of Parasitology, Faculty of Medicine, Chulalongkorn UniversityDepartment of Parasitology, Faculty of Medicine, Chulalongkorn UniversityDepartment of Parasitology, Faculty of Medicine, Chulalongkorn UniversityDepartment of Parasitology, Faculty of Medicine, Chulalongkorn UniversityAbstract Background Culicoides biting midges are known vectors of several pathogens, including arboviruses, protozoa, and filarial worms. Additionally, mermithid nematodes have been found to parasitize Culicoides midges, causing pathogenic effects that reduce host fitness and often lead to death. Consequently, mermithids have potential as biocontrol agents to reduce midge populations. However, the biology of these entomopathogenic nematodes infecting Culicoides in Thailand remains unknown. Methods As part of the leishmaniasis surveillance program in Nakhon Si Thammarat Province, Southern Thailand, we collected Culicoides midges near the residence of a leishmaniasis patient in November 2024. The Culicoides samples were morphologically identified and examined microscopically for parasitic nematodes. Infected midges were dissected to isolate nematodes from each individual. The nematodes were then characterized morphologically using scanning electron microscopy (SEM) and identified molecularly via polymerase chain reaction (PCR) targeting the non-filarial hypervariable region I of the small subunit ribosomal RNA gene (SSU HVR-I) and mermithid small subunit ribosomal RNA (SSU rRNA) gene, followed by nanopore sequencing, phylogenetic analysis, and species delimitation testing. Results A total of 263 field-caught adult Culicoides midges were collected, with Culicoides huffi of the Calvipalpis group being the most abundant species (n = 155, 58.9%). Among these, 35 C. huffi samples, including 4 males, 11 females, and 20 intersex males, were infected with nematodes, resulting in an overall infection rate of 13.3%. The parasitized intersex males, presumably genetically male, exhibited a high degree of feminization in their antennae and wings, which likely enhances female-like behaviors such as detecting and flying toward breeding sites, possibly facilitating parasite transmission and reproductive success. The SEM analysis revealed key morphological features consistent with parasitic nematode larvae of the Mermithidae family, including a long, slender body, a stylet, cephalic papillae, amphids, and a trophosome. Basic Local Alignment Search Tool for nucleotide (BLASTn) analysis of non-filarial SSU HVR-I and mermithid-specific SSU rRNA gene sequences identified all nematodes as mermithids, showing 94.2–94.4% similarity to Pheromermis sp. from the hornet and 97.3% similarity to Mermis sp. from Culicoides obsoletus. Phylogenetic analysis and species delimitation suggest that these sequences represent a single putative species distinct from other known mermithids. Conclusions This study is the first to report mermithid parasitism in Culicoides midges in Thailand, incorporating both ultrastructural and molecular characterization. The novel morphological and molecular insights provided here contribute to a more comprehensive understanding of the biology of entomopathogenic nematodes. Further research is needed to evaluate the potential of these nematodes for the biological control of Culicoides biting midges. Graphical Abstracthttps://doi.org/10.1186/s13071-025-06958-xMermithid parasitismCulicoides huffiBiting midgesIntersexualityBiological controlSouthern Thailand
spellingShingle Chulaluk Promrangsee
Vivornpun Sanprasert
Arunrat Thepparat
Sakone Sunantaraporn
Rinnara Ampol
Rungfar Boonserm
Padet Siriyasatien
Kanok Preativatanyou
First record of mermithid parasitism in adult biting midges, Culicoides huffi (Diptera: Ceratopogonidae), collected from Southern Thailand, with ultrastructural and molecular characterization
Parasites & Vectors
Mermithid parasitism
Culicoides huffi
Biting midges
Intersexuality
Biological control
Southern Thailand
title First record of mermithid parasitism in adult biting midges, Culicoides huffi (Diptera: Ceratopogonidae), collected from Southern Thailand, with ultrastructural and molecular characterization
title_full First record of mermithid parasitism in adult biting midges, Culicoides huffi (Diptera: Ceratopogonidae), collected from Southern Thailand, with ultrastructural and molecular characterization
title_fullStr First record of mermithid parasitism in adult biting midges, Culicoides huffi (Diptera: Ceratopogonidae), collected from Southern Thailand, with ultrastructural and molecular characterization
title_full_unstemmed First record of mermithid parasitism in adult biting midges, Culicoides huffi (Diptera: Ceratopogonidae), collected from Southern Thailand, with ultrastructural and molecular characterization
title_short First record of mermithid parasitism in adult biting midges, Culicoides huffi (Diptera: Ceratopogonidae), collected from Southern Thailand, with ultrastructural and molecular characterization
title_sort first record of mermithid parasitism in adult biting midges culicoides huffi diptera ceratopogonidae collected from southern thailand with ultrastructural and molecular characterization
topic Mermithid parasitism
Culicoides huffi
Biting midges
Intersexuality
Biological control
Southern Thailand
url https://doi.org/10.1186/s13071-025-06958-x
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