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. 2014 Feb 20;8(2):e2688.
doi: 10.1371/journal.pntd.0002688. eCollection 2014 Feb.

Limited dengue virus replication in field-collected Aedes aegypti mosquitoes infected with Wolbachia

Affiliations

Limited dengue virus replication in field-collected Aedes aegypti mosquitoes infected with Wolbachia

Francesca D Frentiu et al. PLoS Negl Trop Dis. .

Abstract

Introduction: Dengue is one of the most widespread mosquito-borne diseases in the world. The causative agent, dengue virus (DENV), is primarily transmitted by the mosquito Aedes aegypti, a species that has proved difficult to control using conventional methods. The discovery that A. aegypti transinfected with the wMel strain of Wolbachia showed limited DENV replication led to trial field releases of these mosquitoes in Cairns, Australia as a biocontrol strategy for the virus.

Methodology/principal findings: Field collected wMel mosquitoes that were challenged with three DENV serotypes displayed limited rates of body infection, viral replication and dissemination to the head compared to uninfected controls. Rates of dengue infection, replication and dissemination in field wMel mosquitoes were similar to those observed in the original transinfected wMel line that had been maintained in the laboratory. We found that wMel was distributed in similar body tissues in field mosquitoes as in laboratory ones, but, at seven days following blood-feeding, wMel densities increased to a greater extent in field mosquitoes.

Conclusions/significance: Our results indicate that virus-blocking is likely to persist in Wolbachia-infected mosquitoes after their release and establishment in wild populations, suggesting that Wolbachia biocontrol may be a successful strategy for reducing dengue transmission in the field.

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Conflict of interest statement

The authors have declared that no competing interests exist.

Figures

Figure 1
Figure 1. Experiment 1: DENV replication in wildtype and field-released (wMel.F) A. aegypti.
DENV replication in bodies (A) and heads (B) of mosquitoes challenged with three strains (DENV2-92T, DENV1-P249, DENV2-P410), assayed at 14 days post-infection. DENV levels determined using one-step qRT-PCR and expressed as copies per 1 μg of total RNA. Bars denote medians. P<0.05 (*), P<0.01 (**), P<0.001 (***). Each point represents an individual mosquito.
Figure 2
Figure 2. Experiment 2: DENV replication in wildtype, outbred laboratory wMel (MGYP2.out) and field-released wMel (wMel.F) A. aegypti.
DENV replication in bodies (A) and heads (B) of mosquitoes challenged with three strains (DENV2-92T, DENV1-P307, DENV3-Cairns08/09), assayed at 14 days post-infection. DENV levels determined using one-step qRT-PCR and expressed as copies per 1 μg of total RNA. Bars denote medians. P<0.05 (*), P<0.01 (**), P<0.001 (***). Each point represents an individual mosquito.
Figure 3
Figure 3. Localization of Wolbachia in different A. aegypti tissues visualized using FISH.
Outbred laboratory wMel (MGYP2.out) (A, C, G, E) and field-released wMel (wMel.F) (B, D, F, H) mosquitoes at day 7 post DENV infection. Wolbachia stained in red (Alexa 594) and cell nuclei in blue (DAPI). Images are representative of 4–5 mosquitoes per line. Bars represent 50 μM scale.
Figure 4
Figure 4. Blood-feeding and Wolbachia densities in whole mosquitoes.
Outbred laboratory wMel (MGYP2.out) and field-released wMel (wMel.F) A. aegypti at 7 (A) and 14 (B) days post blood-feeding (BF) versus non-blood fed (NBF) controls. Bars denote medians. P<0.05 (*), P<0.01 (**), P<0.001 (***). Each point represents an individual mosquito.
Figure 5
Figure 5. Blood-feeding and Wolbachia densities in mosquito heads and bodies.
Bodies (A) and heads (B) of outbred laboratory wMel (MGYP2.out) and field-released wMel (wMel.F) A. aegypti at days 7 and 14 post blood-feeding. Bars denote medians. P<0.05 (*), P<0.01 (**), P<0.001 (***). Each point represents an individual mosquito.

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