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[Preprint]. 2024 Aug 28:rs.3.rs-4756528.
doi: 10.21203/rs.3.rs-4756528/v1.

Symbiotic bacteria Sodalis glossinidius, Spiroplasma sp and Wolbachia do not favour Trypanosoma grayi coexistence in wild population of tsetse flies collected in Bobo-Dioulasso, Burkina Faso

Youssouf Mouliom Mfopit  1 , Etienne Bilgo  2 , Soudah Boma  3 , Martin Bienvenu Somda  3 , Jacques Edounou Gnambani  2 , Maurice Konkobo  2 , Abdoulaye Diabate  2 , Guiguigbaza-Kossigan Dayo  3 , Mohammed Mamman  4 , Soerge Kelm  5 , Emmanuel Oluwadare Balogun  4 , Mohammed Nasir Shuaibu  4 , Junaidu Kabir  4
Affiliations

Symbiotic bacteria Sodalis glossinidius, Spiroplasma sp and Wolbachia do not favour Trypanosoma grayi coexistence in wild population of tsetse flies collected in Bobo-Dioulasso, Burkina Faso

Youssouf Mouliom Mfopit et al. Res Sq. .

Update in

Abstract

Background: Tsetse flies, the biological vectors of African trypanosomes, have established symbiotic associations with different bacteria. Their vector competence is suggested to be affected by bacterial endosymbionts. The current study provided the prevalence of three tsetse symbiotic bacteria and trypanosomes in Glossina species from Burkina Faso.

Results: A total of 430 tsetse flies were captured using biconical traps in four different collection sites around Bobo-Dioulasso (Bama, Bana, Nasso, and Peni), and their guts were removed. Two hundred tsetse were randomly selected and their guts were screened byPCR for the presence of Sodalis glossinidius, Spiroplasmasp., Wolbachia and trypanosomes. Of the 200 tsetse, 196 (98.0%) were Glossina palpalis gambienseand 4 (2.0%) Glossina tachinoides. The overall symbiont prevalence was 49.0%, 96.5%, and 45.0%, respectively for S. glossinidius, Spiroplasma and Wolbachia. Prevalence varied between sampling locations: S. glossinidius(54.7%, 38.5%, 31.6%, 70.8%); Spiroplasma (100%, 100%, 87.7%, 100%); and Wolbachia(43.4%, 38.5%, 38.6%, 70.8%),respectively in Bama, Bana, Nasso and Peni. Noteworthy, no G. tachhnoideswas infected by S. glossinidius and Wolbachia, but they were all infected by Spiroplasma sp. A total of 196 (98.0 %) harbored at least one endosymbionts. Fifty-five (27.5%) carried single endosymbiont. Trypanosomes were found only in G.p. gambiense, but not G. tachinoides. Trypanosomes were present in flies from all study locations with an overall prevalence of 29.5%. In Bama, Bana, Nasso, and Peni, the trypanosome infection rate was respectively 39.6%, 23.1%, 8.8%, and 37.5%. Remarkably, only Trypanosoma grayi was present. Of all trypanosome-infected flies, 55.9%, 98.3%, and 33.9% hosted S. glossinidius, Spiroplasma sp and Wolbachia, respectively. There was no association between Sodalis, Spiroplasma and trypanosome presence, but there was a negative association with Wolbachia presence. We reported1.9 times likelihood of trypanosome absence when Wolbachia was present.

Conclusion: This is the first survey reporting the presence of Trypanosoma grayi in tsetse from Burkina Faso. Tsetse from these localities were highly positive for symbiotic bacteria, more predominantly with Spiroplasma sp. Modifications of symbiotic interactions may pave way for disease control.

Keywords: Burkina Faso; Sodalis glossinidius; Spiroplasma; Trypanosoma grayi; Tsetse flies; Wolbachia.

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Conflict of interest statement

Competing interests: The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Map of study area. Trapping sites are marked with yellow squares. Tsetse flies were collected in Bama (10 traps), Bana (5 traps), Nasso (5 traps) and Peni (10 traps). (Map created with mapchart.net and SW Map).
Figure 2
Figure 2
Phylogenetic analysis of S. glossinidius hemolysin partial gene sequence and its closed relatives. Our isolate is marked by a black diamond. The evolutionary history conducted in MEGA X [33] was inferred by using the Maximum Likelihood method and Kimura 2-parameter model [34]. The tree with the highest log likelihood (−2083.77) is shown. The percentage of trees in which the associated taxa clustered together is shown next to the branches. This analysis involved 12 nucleotide sequences.
Figure 3
Figure 3
Phylogenetic analysis of Spiroplasma16S rRNA partial gene sequence and its closed relatives. Our isolate is marked by a black diamond. The evolutionary history conducted in MEGA X [33] was inferred by using the Maximum Likelihood method and Kimura 2-parameter model [34]. The tree with the highest log likelihood (−927.62) is shown. The percentage of trees in which the associated taxa clustered together is shown next to the branches. This analysis involved 17 nucleotide sequences. There were a total of 307 positions in the final dataset.
Figure 4
Figure 4
Phylogenetic analysis of Wolbachia16S rRNA partial gene sequence and its closed relatives. Our isolate is marked by a black diamond. The evolutionary history conducted in MEGA X [33], was inferred by using the Maximum Likelihood method and Kimura 2-parameter model [34]. The tree with the highest log likelihood (−519.94) is shown. The percentage of trees in which the associated taxa clustered together is shown next to the branches. This analysis involved 12 nucleotide sequences. There were a total of 332 positions in the final dataset.
Figure 5
Figure 5
Coexistence of Sodalis, Spiroplasma, and Wolbachia. Endosymbionts co-existed in 141 tsetse flies. A total of 97 flies carried double infections: Spiroplasma + Wolbachia (43), Sodalis + Wolbachia (1), Sodalis + Spiroplasma (53), and 44 triple infection (Sodalis + Spiroplasma + Wolbachia).
Figure 6
Figure 6
Agarose gel of selected ITS-1 amplicons. Lane M: Molecular Marker 100 bp; Lane 1: T. grayi; Lane 2: T. grayi; Lane 3: T. grayi and kinetoplastid; Lane 4: T. grayi; Lane 5: T. grayi and kinetoplastid; Lane 6: T. grayi; Lane 7: T. grayi and kinetoplastid; Lane P: Positive control; Lane N: Negative control.
Figure 7
Figure 7
Phylogenetic analysis of Trypanosomaand kinetoplastidae ITS-1 gene sequences. Our isolate is marked by a black diamond. The evolutionary history conducted in MEGA X [33], was inferred by using the Maximum Likelihood method and Kimura 2-parameter model [34]. The tree with the highest log likelihood (−1054.41) is shown. The percentage of trees in which the associated taxa clustered together is shown next to the branches. A discrete Gamma distribution was used to model evolutionary rate differences among sites. This analysis involved 14 nucleotide sequences.
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