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Microorganisms
Volume 13
Issue 12
10.3390/microorganisms13122650
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This is an early access version, the complete PDF, HTML, and XML versions will be available soon.
Article

Invertebrate-Derived DNA (iDNA) to Identify Sand Flies’ Bloodmeal: A Molecular Approach to Identifying Hosts in Blood-Feeding Vectors of Leishmaniasis

by
Bruno Oliveira Cova
1,2,*,
Bruno Henrique Saranholi
3,4,
Carla Cristina Gestich
4,
Paulo Roberto Machado
1,2,
Adriano Figueiredo Monte-Alegre
1,2,5 and
Albert Schriefer
1,2,5
1
Serviço de Imunologia, Hospital Universitário Professor Edgard Santos, Universidade Federal da Bahia (UFBA), Salvador 40170-110, BA, Brazil
2
Programa de Pós-Graduação em Ciências da Saúde, Faculdade de Medicina da Bahia, Universidade Federal da Bahia (UFBA), Salvador 40170-110, BA, Brazil
3
Instituto Tecnológico Vale, Belém 66055-090, PA, Brazil
4
Departamento de Genética e Evolução, Universidade Federal de São Carlos, São Carlos 13565-905, SP, Brazil
5
Departamento de Ciências da Biointeração, Instituto de Ciências da Saúde, Universidade Federal da Bahia (UFBA), Salvador 40170-110, BA, Brazil
*
Author to whom correspondence should be addressed.
Microorganisms 2025, 13(12), 2650; https://doi.org/10.3390/microorganisms13122650
Submission received: 30 September 2025 / Revised: 16 November 2025 / Accepted: 17 November 2025 / Published: 21 November 2025
(This article belongs to the Special Issue Interactions between Parasites/Pathogens and Vectors)
Versions Notes

Abstract

DNA metabarcoding data obtained by next generation sequencing (NGS) has been used to identify species in mixed biological samples, such as DNA from the gut content of invertebrates that feed on vertebrates (invertebrate-derived DNA, iDNA). This investigation employed DNA metabarcoding approach to determine vertebrate hosts of female phlebotomine sand flies, blood-feeding leishmaniasis vectors. We evaluated performance across three mitochondrial markers: a mammal-specific mini-barcode (16S rRNA), a pan-vertebrate mini-barcode (12S rRNA), and a standard CytB barcode region. Phlebotomine sand flies collections occurred in the Cacao Region of Southeastern Bahia, Brazil, an American Tegumentary Leishmaniasis (ATL) endemic zone. Our analysis examined iDNA from forty female specimens pooled in thirteen samples of seven sand fly species, including confirmed ATL vectors. Metabarcoding-derived operational taxonomic units (OTUs) underwent taxonomic assignment through comparison with GenBank NCBI® reference databases. Results identified twenty vertebrate OTUs: primates (four OTUs), rodents (four), ungulates (five), marsupials (one), plus a domestic dog and a chicken. Notably, non-mammalian taxa, including reptiles (one OTU) and amphibians (three), were detected. The iDNA metabarcoding approach allowed us to accurately sample the diversity of phlebotomine sandflies’ bloodmeals in a single specimen of a non-engorged female sand fly with mixed feeding.
Keywords: Phlebotominae; bloodmeal; metabarcoding; iDNA; Leishmania Phlebotominae; bloodmeal; metabarcoding; iDNA; Leishmania

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MDPI and ACS Style

Cova, B.O.; Saranholi, B.H.; Gestich, C.C.; Machado, P.R.; Monte-Alegre, A.F.; Schriefer, A. Invertebrate-Derived DNA (iDNA) to Identify Sand Flies’ Bloodmeal: A Molecular Approach to Identifying Hosts in Blood-Feeding Vectors of Leishmaniasis. Microorganisms 2025, 13, 2650. https://doi.org/10.3390/microorganisms13122650

AMA Style

Cova BO, Saranholi BH, Gestich CC, Machado PR, Monte-Alegre AF, Schriefer A. Invertebrate-Derived DNA (iDNA) to Identify Sand Flies’ Bloodmeal: A Molecular Approach to Identifying Hosts in Blood-Feeding Vectors of Leishmaniasis. Microorganisms. 2025; 13(12):2650. https://doi.org/10.3390/microorganisms13122650

Chicago/Turabian Style

Cova, Bruno Oliveira, Bruno Henrique Saranholi, Carla Cristina Gestich, Paulo Roberto Machado, Adriano Figueiredo Monte-Alegre, and Albert Schriefer. 2025. "Invertebrate-Derived DNA (iDNA) to Identify Sand Flies’ Bloodmeal: A Molecular Approach to Identifying Hosts in Blood-Feeding Vectors of Leishmaniasis" Microorganisms 13, no. 12: 2650. https://doi.org/10.3390/microorganisms13122650

APA Style

Cova, B. O., Saranholi, B. H., Gestich, C. C., Machado, P. R., Monte-Alegre, A. F., & Schriefer, A. (2025). Invertebrate-Derived DNA (iDNA) to Identify Sand Flies’ Bloodmeal: A Molecular Approach to Identifying Hosts in Blood-Feeding Vectors of Leishmaniasis. Microorganisms, 13(12), 2650. https://doi.org/10.3390/microorganisms13122650

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