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. 2025 May 22;15(1):17732.
doi: 10.1038/s41598-025-97288-6.

Unstable laboratory Wolbachia strain w-Anga is negatively correlated with Plasmodium falciparum in wild malaria vectors

Dembélé L Estelle #  1   2   3 , Gnambani E Jacques  1   4   3 , Saré Issiaka  1   4 , Brian Lovett  5 , Diabaté Abdoulaye  6   7   8 , Bilgo Etienne  9   10
Affiliations

Unstable laboratory Wolbachia strain w-Anga is negatively correlated with Plasmodium falciparum in wild malaria vectors

Dembélé L Estelle et al. Sci Rep. .

Abstract

Spread of insecticides resistance threatens the control of malaria. In this context, biological control using an endosymbiotic bacterium Wolbachia is being explored as a complementary method for its control. However, for optimal use of this bacterium in biocontrol strategies, it is imperative to characterize it. So, Anopheles gambiae complex mosquitoes were collected, morphologically identified, then blood fed and gravid female mosquitoes oviposited individually. After oviposition, the species of parent was molecularly determined, along with their w-Anga infection status. Additionally, we performed 16SrRNA gene sequencing of w-Anga-positive mosquitoes to determine their phylogeny. Finally, we amplified gene encoding the circumsporozoite protein to determinate their Plasmodium falciparum infection status and assessed the stability of w-Anga transmission of positive females and their offspring. From the results obtained, our w-Anga strains cluster with other Wolbachia Supergroup B strains. However, the prevalence of Plasmodium falciparum infection was lower in Wolbachia-infected females (4.59%) than in those uninfected (22.02%). Furthermore, the transmission frequency of this bacterium in infected Anopheles coluzzii females of the F0 generation to F1 offspring was 10.64% and 16.67% from infected females of the F1 generation to F2 offspring. This study results will serve as preliminary data for the possible use of Wolbachia in malaria control.

Keywords: Anopheles gambiae complex; Plasmodium falciparum; w-Anga; Malaria; Phylogeny; Stability.

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Conflict of interest statement

Declarations. Competing interests: The authors declare no competing financial interests. Ethical approval: Experiments with animals were carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. In addition, Experiments followed the IRSS Animal Welfare Assurance A5926-01. Trained personnel and veterinarians cared for animals involved in this study and all efforts were made to minimize suffering. All work with w-Anga was performed under biosafety containment level II requirements. Consent for publication: All authors have approved the final manuscript and consent for the publication.

Figures

Fig. 1
Fig. 1
Mosquito samples collected have been to 16SrRNA Nested PCR to obtain amplicons positive for Wolbachia strain w-Anga. These amplicons underwent 16SrRNA gene sequencing by Sanger technology using GENEWIZ/AZENTA’s internal formulation of BigDye V3 chemistry on an ABI3730xl sequencer. The sequences obtained were subjected to phylogenetic analysis, which involved aligning them with data available in the GenBank database using BLAST. Following the BLAST alignment, we established a phylogenomic relationship between our samples (five strains, including four derived from our 16SrRNA amplicons and a positive control of wAlbB strain) and 33 Wolbachia strains from NCBI by constructing a phylogenetic tree using the “Fasttree” program in the “Jupyter Notebook” application, version 4.2.4. This phylogenetic tree was visualized and annotated using “ITOL v6.9.1.” The assembly names on the phylogenetic tree were color-coded based on the identity of the supergroups (A-F) and other 16SrRNA sequences.
Fig. 2
Fig. 2
Plasmodium falciparum infection status based on w-Anga infection within wild caught Anopheles coluzzii. The proportions of Anopheles coluzzii mosquito females infected with Plasmodium among w-Anga-positive females (5/109) and among w-Anga-negative females (24/109) are shown in red (with a statistically significant difference between the two proportions, P = 0.0003309).
Fig. 3
Fig. 3
Impact of natural w-Anga infections on fecundity and fertility of wild caught Anopheles coluzzii mosquitoes. (A) Average number of eggs uninfected and infected to w-Anga. (B) Average number of larvae uninfected and infected to w-Anga.
Fig. 4
Fig. 4
Hauts Bassins region in Burkina Faso representation, in which the two collection sites Bama and Soumousso are located in red dots.
See this image and copyright information in PMC

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