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. 2016 Sep;111(9):577-87.
doi: 10.1590/0074-02760160238. Epub 2016 Aug 25.

Effects of environment, dietary regime and ageing on the dengue vector microbiota: evidence of a core microbiota throughout Aedes aegypti lifespan

Mariana Rocha David  1 , Lilha Maria Barbosa Dos Santos  1 , Ana Carolina Paulo Vicente  2 , Rafael Maciel-de-Freitas  1
Affiliations

Effects of environment, dietary regime and ageing on the dengue vector microbiota: evidence of a core microbiota throughout Aedes aegypti lifespan

Mariana Rocha David et al. Mem Inst Oswaldo Cruz. 2016 Sep.

Abstract

Mosquito midgut microbiota is a key component of vector competence, as gut bacteria can disturb pathogen development. In this study, we addressed the microbiota composition of Aedes aegypti during its lifespan, under field conditions. We also investigated the possible effects of environment, dietary regime and ageing on the gut community composition. We employed culture independent and dependent approaches to characterise vector microbiota. There was evidence of a lifelong stable core microbiota after mosquitoes were released into an urban settlement, where they presumably fed on a range of vertebrate hosts and carbohydrate sources. This core was formed mainly of bacteria belonging to the genera Pseudomonas, Acinetobacter, Aeromonas and Stenotrophomonas and to the families Oxalobacteraceae, Enterobacteriaceae and Comamonadaceae. We showed that both dietary regime and age were associated with the abundance of some bacterial groups in the Ae. aegypti microbiota. The majority of the bacterial groups we identified have been detected in the midgut of Ae. aegypti from laboratory and wild populations, indicating a possible core microbiota associated with this mosquito species. Our findings suggest that Ae. aegypti harbours a stable bacterial community during its adult life, similar to mosquito populations from distinct geographic areas, which may be further explored for arbovirus biocontrol strategies.

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Figures

Fig. 1
Fig. 1. : rarefaction curves for each sample. Operational taxonomic units (OTUs) at 3% distance were plotted for each sample against the number of sequences generated through 454 pyrosequencing (Sample size). SFY: sugar-fed young; BFY: blood-fed young; SFO: sugar-fed old; DAR: days after release in the mark-release-recapture experiment; WD: wild.
Fig. 2
Fig. 2. : principal coordinate analysis (PCoA) of Bray-Curtis distances among Aedes aegypti microbiota samples. Bray-Curtis distance was calculated with operational taxonomic units at 3% maximum distance. SFY: sugar-fed young (yellow); BFY: blood-fed young (pink); SFO: sugar-fed old (blue); MRR: mark-released-recaptured (orange); WD: wild (black). Each axis shows percentage of variation explained.
Fig. 3
Fig. 3. : Aedes aegypti microbiota composition per sample. Gut bacterial taxonomic composition of Ae. aegypti adult females was determined via 16S rRNA gene deep sequencing. SFY: sugar-fed young; BFY: blood-fed young; SFO: sugar-fed old; DAR: days after release in the mark-release-recapture experiment; WD: wild. Taxa with < 2% relative abundance were pooled as “Other”; *: not discriminated at genus level.
Fig. 4
Fig. 4. : phylogenetic tree of bacterial taxa detected in Aedes aegypti female midgut via 16S rRNA gene deep sequencing. Yellow circles indicate taxa identified in sugar-fed young females (SFY); pink circles in blood-fed young (BFY); light blue in sugar-fed old (SFO); orange in mark-released-recaptured (MRR); and black in wild (WD) females. Taxa were classified according to their phyla: Bacteroidetes (blue), Actinobacteria (green), Firmicutes (purple), and Proteobacteria (red); *: not discriminated at genus level.
Fig. 5
Fig. 5. : Venn diagram showing the number of bacterial taxa identified in the Aedes aegypti microbiota via 16S rRNA gene deep sequencing. SFY: sugar-fed young; BFY: blood-fed young; SFO: sugar-fed old; MRR: mark-released-recaptured; WD: wild.
Fig. 6
Fig. 6. : Aedes aegypti microbiota composition per group. Gut bacterial taxonomic composition of Ae. aegypti adult females was determined via 16S rRNA gene deep sequencing. SFY: sugar-fed young; BFY: blood-fed young; SFO: sugar-fed old; MRR: mark-released-recaptured; WD: wild. Taxa with < 2% relative abundance were pooled as “Other”; *: not discriminated at genus level.
Fig. 7
Fig. 7. : predicted functions of the bacterial communities found in Aedes aegypti adult females. Yellow bars indicate Kyoto Encyclopedia of Genes and Genomes metabolic pathways predicted in the microbiota of sugar-fed young females (SFY); pink bars in blood-fed young (BFY); light blue in sugar-fed old (SFO); orange in mark-released-recaptured (MRR); and black in wild females (WD). *: indicates gene categories significantly different between SFO and WD mosquitoes; **: indicates gene categories significantly different among SFO and both MRR and WD mosquitoes (Kruskal-Wallis test, Benjamini-Hochberg corrected p-value < 0.05).
Fig. 8
Fig. 8. : phylogenetic tree of culturable bacterial genera recovered from Aedes aegypti females. SFY: sugar-fed young (yellow); BFY: blood-fed young (pink); SFO: sugar-fed old (blue); MRR: mark-released-recaptured (orange). Bacterial genera were classified according to their phyla: Bacteroidetes (blue), Actinobacteria (green) and Proteobacteria (red).
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