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Species Composition of Bacterial Communities Influences Attraction of Mosquitoes to Experimental Plant Infusions

  • Microbiology of Aquatic Systems
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Abstract

In the container habitats of immature mosquitoes, catabolism of plant matter and other organic detritus by microbial organisms produces metabolites that mediate the oviposition behavior of Aedes aegypti and Aedes albopictus. Public health agencies commonly use oviposition traps containing plant infusions for monitoring populations of these mosquito species, which are global vectors of dengue viruses. In laboratory experiments, gravid females exhibited significantly diminished responses to experimental infusions made with sterilized white oak leaves, showing that attractive odorants were produced through microbial metabolic activity. We evaluated effects of infusion concentration and fermentation time on attraction of gravid females to infusions made from senescent bamboo or white oak leaves. We used plate counts of heterotrophic bacteria, total counts of 4′,6-diamidino-2-phenylindole-stained bacterial cells, and 16S ribosomal DNA (rDNA) polymerase chain reaction–denaturing gradient gel electrophoresis (DGGE) to show that changes in the relative abundance of bacteria and the species composition of bacterial communities influenced attraction of gravid A. aegypti and A. albopictus mosquitoes to infusions. DGGE profiles showed that bacterial species composition in infusions changed over time. Principal components analysis indicated that oviposition responses to plant infusions were in general most affected by bacterial diversity and abundance. Analysis of bacterial 16S rDNA sequences derived from DGGE bands revealed that Proteobacteria (Alpha-, Beta-, Delta-, and Gamma-) were the predominant bacteria detected in both types of plant infusions. Gravid A. aegypti were significantly attracted to a mix of 14 bacterial species cultured from bamboo leaf infusion. The oviposition response of gravid mosquitoes to plant infusions is strongly influenced by abundance and diversity of bacterial species, which in turn is affected by plant species, leaf biomass, and fermentation time.

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Acknowledgments

Partial support for our research was provided by the NIH-NIAID through cooperative agreement U01-AI-58303-01, the Blanton J. Whitmire endowment at North Carolina State University, and the Bill and Melinda Gates Foundation. We thank Luma Abu Ayyash for excellent assistance in the laboratory.

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Authors and Affiliations

  1. Department of Entomology, North Carolina State University, Campus Box 7613, Raleigh, NC, 27695-7613, USA

    Loganathan Ponnusamy, Coby Schal & Charles S. Apperson

  2. Department of Tropical Medicine, Tulane University, New Orleans, LA, 70112-2699, USA

    Dawn M. Wesson

  3. Department of Statistics, North Carolina State University, Campus Box 8203, Raleigh, NC, 27695-8203, USA

    Consuelo Arellano

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  1. Loganathan Ponnusamy
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  2. Dawn M. Wesson
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  3. Consuelo Arellano
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  4. Coby Schal
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  5. Charles S. Apperson
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Correspondence to Charles S. Apperson.

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Supplemental Table 1

Bacterial phylotypes identified* from predominant 16S rDNA-DGGE bands from BL infusions. Phylogenetic affiliations were determined based on classification of band sequences in a neighbor-joining phylogenetic tree (DOC 50 kb)

Supplemental Table 2

Bacterial phylotypes identified* from predominant 16S rDNA-DGGE bands from WOL infusions. Phylogenetic affiliations were determined based on classification of band sequences in a neighbor-joining phylogenetic tree (DOC 60 kb)

Supplemental Table 3

Results of mixed-model ANOVA of effects of trial and fermentation time (day) on bacterial cell growth and diversity in BL infusions (DOC 38 kb)

Supplemental Table 4

Effects of infusion concentration§ and fermentation time on bacterial species diversity in WOL infusions (DOC 37 kb)

Supplemental Table 5

Correlation matrix for infusion-associated variables used in principal components analysis (DOC 29.5 kb)

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Ponnusamy, L., Wesson, D.M., Arellano, C. et al. Species Composition of Bacterial Communities Influences Attraction of Mosquitoes to Experimental Plant Infusions. Microb Ecol 59, 158–173 (2010). https://doi.org/10.1007/s00248-009-9565-1

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