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Comparative analysis of midgut bacterial communities of Aedes aegypti mosquito strains varying in vector competence to dengue virus

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Abstract

Differences in midgut bacterial communities of Aedes aegypti, the primary mosquito vector of dengue viruses (DENV), might influence the susceptibility of these mosquitoes to infection by DENV. As a first step toward addressing this hypothesis, comparative analysis of bacterial communities from midguts of mosquito strains with differential genetic susceptibility to DENV was performed. 16S rRNA gene libraries and real-time PCR approaches were used to characterize midgut bacterial community composition and abundance in three Aedes aegypti strains: MOYO, MOYO-R, and MOYO-S. Although Pseudomonas spp.-related clones were predominant across all libraries, some interesting and potentially significant differences were found in midgut bacterial communities among the three strains. Pedobacter sp.- and Janthinobacterium sp.-related phylotypes were identified only in the MOYO-R strain libraries, while Bacillus sp. was detected only in the MOYO-S strain. Rahnella sp. was found in MOYO-R and MOYO strains libraries but was absent in MOYO-S libraries. Both 16S rRNA gene library and real-time PCR approaches confirmed the presence of Pedobacter sp. only in the MOYO-R strain. Further, real-time PCR-based quantification of 16S rRNA gene copies showed bacterial abundance in midguts of the MOYO-R strain mosquitoes to be at least 10–100-folds higher than in the MOYO-S and MOYO strain mosquitoes. Our study identified some putative bacteria with characteristic physiological properties that could affect the infectivity of dengue virus. This analysis represents the first report of comparisons of midgut bacterial communities with respect to refractoriness and susceptibility of Aedes aegypti mosquitoes to DENV and will guide future efforts to address the potential interactive role of midgut bacteria of Aedes aegypti mosquitoes in determining vectorial capacity for DENV.

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Authors and Affiliations

  1. Molecular Biology Unit, National Centre for Cell Science, Ganeshkhind, Pune, Maharashtra, India

    Shakti S. Charan, Kiran D. Pawar, Milind S. Patole & Yogesh S. Shouche

  2. Eck Institute for Global Health Division of Biological Sciences, University of Notre Dame, Notre Dame, IN, USA

    David W. Severson

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  1. Shakti S. Charan
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  2. Kiran D. Pawar
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  3. David W. Severson
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  4. Milind S. Patole
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  5. Yogesh S. Shouche
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Correspondence to Yogesh S. Shouche.

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Supplementary Fig. 1

Neighbor-joining tree constructed from sequences affiliated to Pseudomonas sp. from 16S rRNA gene libraries of MOYO-R, MOYO-S, and MOYO strains. The tree was generated by using the neighbor joining method with Kimura 2 parameter distances in MEGA 4.0 software. Numbers at nodes indicate percent bootstrap values above 50 (1,000 replicates). The bar indicates the Jukes–Cantor evolutionary distance. In the sequence codes, first letters R, S, and M indicate MOYO-R, MOYO-S, and MOYO strain, respectively. (DOC 74 kb)

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Charan, S.S., Pawar, K.D., Severson, D.W. et al. Comparative analysis of midgut bacterial communities of Aedes aegypti mosquito strains varying in vector competence to dengue virus. Parasitol Res 112, 2627–2637 (2013). https://doi.org/10.1007/s00436-013-3428-x

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