Metagenomic exploration of the bacteriome reveals natural Wolbachia infections in yellow fever mosquito Aedes aegypti and Asian tiger mosquito Aedes albopictus

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Abstract

Background

Dengue and associated complications are spreading to non-endemic regions of Pakistan. Vector control, the foremost and widely adopted strategy for managing dengue has been implemented through various measures in Pakistan. Biological control through the use of Wolbachia , a bacterium naturally present in various insect genera, including Aedes , has demonstrated promising results globally. In this study we collected Aedes species and investigated their microbiomes with a particular focus on identifying the endosymbiont Wolbachia .

Methods

Mosquitoes were collected via Gravitraps in the Peshawar region of northwest Pakistan. The identity of the mosquitoes was initially confirmed through morphological characters followed by molecular identification using species-specific Cytochrome oxidase I (COI) primers. The DNA from female Ae. aegypti and Ae. albopictus was further subjected to 16S rRNA sequencing. The hypervariable regions V3/V4 of 16S rRNA were used for sequencing using the paired-end Illumina MiSeq platform.

Results

The phylogenetic analysis of the COI gene in our samples demonstrated similarity to Aedes species previously documented in Pakistan. In comparative analysis of their microbiomes, Ae. albopictus was found to harbor 921 bacterial species, while Ae. aegypti only had 239 species. The metagenomic analysis revealed Wolbachia pipientis infection in Ae. aegypti while co-infection of Wolbachia pipientis and Wolbachia bourtzisii was detected in Ae. albopictus microbiota.

Conclusion

Both Ae. aegypti and Ae. albopictus are present in Peshawar region of Khyber Pakhtoonkhwa province of Pakistan. Comparative analysis of the bacteriome showed higher bacterial diversity for Ae. albopictus as compared to Ae. aegypti. The investigation revealed Wolbachia infection in both Aedes mosquitoes species.

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