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. 2023 Apr 25;16(1):142.
doi: 10.1186/s13071-023-05745-w.

Screening of natural Wolbachia infection in mosquitoes (Diptera: Culicidae) from the Cape Verde islands

Aires Januário Fernandes da Moura  1   2 , Vera Valadas  3 , Silvania Da Veiga Leal  4 , Eddyson Montalvo Sabino  3   5 , Carla A Sousa  3 , João Pinto  3
Affiliations

Screening of natural Wolbachia infection in mosquitoes (Diptera: Culicidae) from the Cape Verde islands

Aires Januário Fernandes da Moura et al. Parasit Vectors. .

Abstract

Background: Wolbachia pipientis is an endosymbiont bacterium that induces cytoplasmic incompatibility and inhibits arboviral replication in mosquitoes. This study aimed to assess Wolbachia prevalence and genetic diversity in different mosquito species from Cape Verde.

Methods: Mosquitoes were collected on six islands of Cape Verde and identified to species using morphological keys and PCR-based assays. Wolbachia was detected by amplifying a fragment of the surface protein gene (wsp). Multilocus sequence typing (MLST) was performed with five housekeeping genes (coxA, gatB, ftsZ, hcpA, and fbpA) and the wsp hypervariable region (HVR) for strain identification. Identification of wPip groups (wPip-I to wPip-V) was performed using PCR-restriction fragment length polymorphism (RFLP) assay on the ankyrin domain gene pk1.

Results: Nine mosquito species were collected, including the major vectors Aedes aegypti, Anopheles arabiensis, Culex pipiens sensu stricto, and Culex quinquefasciatus. Wolbachia was only detected in Cx. pipiens s.s. (100% prevalence), Cx. quinquefasciatus (98.3%), Cx. pipiens/quinquefasciatus hybrids (100%), and Culex tigripes (100%). Based on the results of MLST and wsp hypervariable region typing, Wolbachia from the Cx. pipiens complex was assigned to sequence type 9, wPip clade, and supergroup B. PCR/RFLP analysis revealed three wPip groups in Cape Verde, namely wPip-II, wPip-III, and wPip-IV. wPip-IV was the most prevalent, while wPip-II and wPip-III were found only on Maio and Fogo islands. Wolbachia detected in Cx. tigripes belongs to supergroup B, with no attributed MLST profile, indicating a new strain of Wolbachia in this mosquito species.

Conclusions: A high prevalence and diversity of Wolbachia was found in species from the Cx. pipiens complex. This diversity may be related to the mosquito's colonization history on the Cape Verde islands. To the best of our knowledge, this is the first study to detect Wolbachia in Cx. tigripes, which may provide an additional opportunity for biocontrol initiatives.

Keywords: Cape Verde; Culex pipiens; Culex tigripes; Genotyping; Mosquitoes; Wolbachia.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
Map of the North Atlantic region showing the geographic location of the Cape Verde islands. Mosquito samples were collected on the islands of Santo Antão, Boavista, Maio, Santiago, Fogo, and Brava (highlighted in black)
Fig. 2
Fig. 2
Phylogenetic tree generated from concatenated sequences of MLST loci (coxA, gatB, ftsZ, fbpA, hcpA) and the wsp hypervariable region. Numbers on branches indicate percentage bootstrap support (1000 replicates). Reference sequences were obtained from the Wolbachia MLST database and are marked by full circles. Each Wolbachia supergroup is marked with a different color: yellow, supergroup B; black, supergroup A; red, supergroup D; and green, supergroup F. The scale bar indicates the number of substitutions
Fig. 3
Fig. 3
Phylogenetic tree generated from pk1 sequences by Bayesian analysis. Known wPip group pk1 sequences are marked by full circles. Numbers on branches indicate percentage bootstrap support (1000 replicates). The scale bar indicates the number of substitutions
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