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. 2019:26:5.
doi: 10.1051/parasite/2019005. Epub 2019 Feb 7.

Molecular identification of Wolbachia and Sodalis glossinidius in the midgut of Glossina fuscipes quanzensis from the Democratic Republic of Congo

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Molecular identification of Wolbachia and Sodalis glossinidius in the midgut of Glossina fuscipes quanzensis from the Democratic Republic of Congo

Gustave Simo et al. Parasite. 2019.

Abstract

During the last 30 years, investigations on the microbiome of different tsetse species have generated substantial data on the bacterial flora of these cyclical vectors of African trypanosomes, with the overarching goal of improving the control of trypanosomiases. It is in this context that the presence of Wolbachia and Sodalis glossinidius was studied in wild populations of Glossina fuscipes quanzensis from the Democratic Republic of Congo. Tsetse flies were captured with pyramidal traps. Of the 700 Glossina f. quanzensis captured, 360 were dissected and their midguts collected and analyzed. Sodalis glossinidius and Wolbachia were identified by PCR. The Wolbachia-positive samples were genetically characterized with five molecular markers. PCR revealed 84.78% and 15.55% midguts infected by Wolbachia and S. glossinidius, respectively. The infection rates varied according to capture sites. Of the five molecular markers used to characterize Wolbachia, only the fructose bis-phosphate aldolase gene was amplified for about 60% of midguts previously found with Wolbachia infections. The sequencing results confirmed the presence of Wolbachia and revealed the presence of S. glossinidius in the midgut of Glossina f. quanzensis. A low level of midguts were naturally co-infected by both bacteria. The data generated in this study open a framework for investigations aimed at understanding the contribution of these symbiotic microorganisms to the vectorial competence of Glossina fuscipes quanzensis.

Au cours des 30 dernières années, les recherches sur le microbiome de différentes espèces de glossines ont généré des données substantielles sur la flore bactérienne de ces vecteurs cycliques des trypanosomes africains, l’objectif principal étant d’améliorer le contrôle des trypanosomiases. C’est dans cette optique que la présence de Wolbachia et Sodalis glossinidius a été étudiée dans des populations sauvages de Glossina fuscipes quanzensis de la République démocratique du Congo. Les glossines ont été capturées avec des pièges pyramidaux. Parmi les 700 Glossina f. quanzensis capturés, 360 ont été disséqués et leur estomac récupéré et analysé. Sodalis glossinidius et Wolbachia ont été identifiés par PCR. Les échantillons positifs pour Wolbachia ont été génétiquement caractérisés avec cinq marqueurs moléculaires. La PCR a révélé que 84,78 % et 15,55 % de l’intestin moyen étaient respectivement infectés par Wolbachia et S. glossinidius. Les taux d’infection variaient selon les sites de capture. Sur les cinq marqueurs moléculaires utilisés pour caractériser Wolbachia, seul le gène de la fructose bis-phosphate aldolase a été amplifié pour environ 60 % des intestins moyens précédemment identifiés porteurs de Wolbachia. Les résultats du séquençage ont confirmé la présence de Wolbachia et ont révélé la présence de S. glossinidius dans l’intestin moyen de Glossina f. quanzensis. Un faible taux des intestins moyens était naturellement co-infecté par les deux bactéries. Les données de cette étude ouvrent un cadre pour des recherches visant à comprendre la contribution de ces microorganismes symbiotiques sur la compétence vectorielle de Glossina fuscipes quanzensis.

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Figures

Figure 1
Figure 1
Map showing areas where tsetse flies were caught (stars).
Figure 2
Figure 2
Agarose gel showing the DNA profile resulting from the molecular identification of S. glossinidius using pSG2 primers. Lane 1: negative control; lane 9: positive control; lanes 2, 4 and 7: samples positive for S. glossinidius; Lane 3, 5, 6 and 8: samples negative for S. glossinidius. MW: 100 bp DNA ladder.
Figure 3
Figure 3
Agarose gel illustrating the DNA profile resulting from the amplification of a DNA fragment of the Wolbachia surface protein gene using wsp primers. MW: 1 kb DNA ladder molecular weight marker; lane 1: negative control; lane 11: positive control; Lane 2, 3, 6 and 9: samples positive for Wolbachia; lanes 4, 5, 7, 8 and 10: samples negative for Wolbachia.

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