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. 2025 Jun 9;18(1):216.
doi: 10.1186/s13071-025-06860-6.

In vitro interaction profiles and midgut glycoconjugates of Trichophoromyia spp./Leishmania (Viannia) lainsoni

Thais Gouvea de Morais  1   2 , Yetsenia Del Valle Sánchez Uzcátegui  1   2   3 , Chubert Bernardo Castro de Sena  4 , Rodrigo Ribeiro Furtado  1   2 , Fernando Tobias Silveira  2 , Edilene Oliveira da Silva #  1   4 , Thiago Vasconcelos Dos Santos #  5   6
Affiliations

In vitro interaction profiles and midgut glycoconjugates of Trichophoromyia spp./Leishmania (Viannia) lainsoni

Thais Gouvea de Morais et al. Parasit Vectors. .

Abstract

Background: Knowledge of sand fly-Leishmania attachment determinants is pivotal for providing evidence on vector status. Considering the Amazonian transmission context of Trichophoromyia spp.-L. (Viannia) lainsoni, the present study aimed to assess in vitro interactions and detect gut glycoconjugates associated with this vector-parasite association.

Methods: Field-caught Trichophoromyia brachipyga and Trichophoromyia ubiquitalis were tested. Lutzomyia longipalpis reared in the laboratory was used as a control. The intestines were obtained by dissection, and the species were confirmed by morphology. Interactions for each sand fly-Leishmania association were individually performed via an in vitro incubation system. N-acetyl-D-glucosamine (GlcNAc), galactose-(β 1,3)-GalNAc (Gal/GalNAc), and N-acetyl-D-galactosamine (GalNAc) glycoconjugates of Trichophoromyia spp. were analyzed by Western blotting using corresponding peroxidase-conjugated lectins.

Results: No difference was found between Th. ubiquitalis and Lu. longipalpis attachment with L. (V.) lainsoni, and Lu. longipalpis with Leishmania (Leishmania) infantum (control). However, Th. brachipyga-L. (V.) lainsoni attachment was weaker than that of the control. Trichophoromyia spp. were negative for residues of GlcNAc terminally exposed. Trichophoromyia ubiquitalis was positive for GalNAc and Gal/GalNAc, whereas Th. brachipyga presented only residues of GalNac terminally exposed.

Conclusions: The present study suggests that Trichophoromyia spp. sand flies, particularly Th. brachipyga, are susceptible to L. (V.) lainsoni, based on the observed vector-parasite attachment profiles and detection of GalNAc in their midguts. This supports early field data suggesting the vector status of these sand fly species.

Keywords: Leishmania; Attachment; Glycoconjugates; Sand fly; Vector.

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Conflict of interest statement

Declarations. Ethics approval and consent to participate: The capture and processing of invertebrate fauna (Diptera: Psychodidae) were authorized by the Sistema de Autorização e Informação em Biodiversidade (SISBio) under protocol no. 89906–1. Consent for publication: Not applicable. Competing interests: The authors declare no competing interests.

Figures

Fig. 1
Fig. 1
In vitro interactions between Trichophoromyia brachipyga–Leishmania (Viannia) lainsoni, Th. ubiquitalis–L. (V.) lainsoni, and Lutzomyia longipalpis–L. (V.) lainsoni. The susceptible association Lu. longipalpis–L. (L.) infantum was used as a control. The attachment profiles were classified into three categories: weak (less than 100 parasites per gut), moderate (100–1000 parasites per gut), and heavy (more than 1000 parasites per gut); values inside the bars indicate percentages of each category; values on the bars indicate the number of guts assessed. *P < 0.05
Fig. 2
Fig. 2
Western blot and densitometry analyses of peroxidase-conjugated lectin reaction profiles for the detection of sand fly gut glycoconjugates. A Peanut agglutinin (PNA), indicating galactose/GalNAc (Gal/GalNAc); B Helix pomatia agglutinin (HPA), indicating N-acetyl-d-galactosamine (GalNac). 1 indicates Trichophoromyia ubiquitalis; 2 indicates Th. brachipyga; 3 indicates Lutzomyia longipalpis. WM, molecular weight marker (values in kilodaltons)
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