Application of biomolecular techniques on tsetse fly puparia for species identification at larvipostion sites

Gimonneau Geoffrey, Hounyeme Robert Eustache, Quartey Myra, Barry Issiaka, Ravel Sophie, Boulange Alain. 2024. Application of biomolecular techniques on tsetse fly puparia for species identification at larvipostion sites. Bulletin of Entomological Research, 7 p.

Liste HCERES des revues (en SHS) : oui

Thème(s) HCERES des revues (en SHS) : Psychologie-éthologie-ergonomie

Résumé : Puparia are commonly found in tsetse fly larviposition sites during studies on larval ecology. This chitinous shell is representative of past or ongoing exploitation of these sites by tsetse flies. The morphological characteristics of the puparium are not sufficiently distinctive to allow identification of the species. This study explores the applicability of biomolecular techniques on empty puparia for tsetse fly species identification. Five techniques were compared for DNA extraction from tsetse fly puparia, 1/Chelex® 100 Resin, 2/CTAB, 3/Livak's protocol, 4/DEB + proteinase K and 5/QIAamp® DNA Mini kit, using two homogenisation methods (manual and automated). Using a combination of two primer pairs, Chelex, CTAB, and DEB + K proved the most efficient on fresh puparia with 90, 85, and 70% samples identified, respectively. Shifting from fresh to one- to nine-month-old puparia, the Chelex method gave the best result allowing species identification on puparia up to seven months old. The subsequent testing of the Chelex extraction protocol identified 152 (60%) of 252 field-collected puparia samples at species level. The results show that reliable genetic identification of tsetse flies species can be performed from empty puparia, what can prove of great interest for future ecological studies on larviposition sites. The Chelex technique was the most efficient for DNA extraction, though the age-limit of the samples stood at seven months, beyond which DNA degradation probably compromises the genetic analysis.

Mots-clés Agrovoc : identification, écologie animale, pupe, Glossina palpalis, biologie moléculaire, Glossinidae

Mots-clés géographiques Agrovoc : Burkina Faso

Mots-clés libres : DNA extraction, Larviposition, Molecular diagnosis, Puparium, Tsetse fly ecology

Classification Agris : L72 - Organismes nuisibles des animaux
L20 - Écologie animale

Champ stratégique Cirad : CTS 4 (2019-) - Santé des plantes, des animaux et des écosystèmes

Agences de financement européennes : European Commission

Agences de financement hors UE : Bill and Melinda Gates Foundation

Programme de financement européen : H2020

Projets sur financement : (EU) COntrolling and progressively Minimizing the Burden of Animal Trypanosomosis, (USA) Trypa-NO !

Auteurs et affiliations

• Gimonneau Geoffrey, CIRAD-BIOS-UMR INTERTRYP (SEN) ORCID: 0000-0002-0613-841X - auteur correspondant
• Hounyeme Robert Eustache, CIRAD-BIOS-UMR INTERTRYP (FRA)
• Quartey Myra, CIRDES (BFA)
• Barry Issiaka, CIRDES (BFA)
• Ravel Sophie, IRD (FRA)
• Boulange Alain, CIRAD-BIOS-UMR INTERTRYP (CIV) ORCID: 0000-0002-8505-7465

Source : Cirad-Agritrop (https://agritrop.cirad.fr/608787/)

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