<b>The effect of repeat feeding on dengue virus transmission potential in </b><b><i>Wolbachia</i></b><b>-infected </b><b><i>Aedes aegypti</i></b><b> following extended egg quiescence</b>

As <i>Wolbachia pipientis</i> is more widely being released into field populations of <i>Aedes aegypti</i> for disease control, the ability to select the appropriate strain for differing environments is increasingly important. A previous study revealed that longer-term quiescence in the egg phase reduced the fertility of mosquitoes, especially those harboring the <i>w</i>AlbB <i>Wolbachia</i> strain. This infertility was also associated with a greater biting rate. Here, we attempt to quantify the effect of this heightened biting behavior on the transmission potential of the dengue virus using a combination of assays for fitness, probing behavior, and vector competence, allowing repeat feeding, and incorporate these effects in a model of R₀. We show that <i>Wolbachia</i>-infected infertile mosquitoes are more interested in feeding almost immediately after an initial blood meal relative to wild type and <i>Wolbachia</i>-infected fertile mosquitoes and that these differences continue for up to 8 days over the period we measured. As a result, the infertile <i>Wolbachia</i> mosquitoes have higher virus prevalence and loads than <i>Wolbachia</i>-fertile mosquitoes. We saw limited evidence of <i>Wolbachia</i>-mediated blocking in the disseminated tissue (legs) in terms of prevalence but did see reduced viral loads. Using a previously published estimate of the extrinsic incubation period, we demonstrate that the effect of repeat feeding/infertility is insufficient to overcome the effects of <i>Wolbachia</i>-mediated blocking on R₀. These estimates are very conservative, however, and we posit that future studies should empirically measure EIP under a repeat feeding model. Our findings echo previous work where periods of extensive egg quiescence affected the reproductive success of <i>Wolbachia</i>-infected <i>Ae. aegypti</i>. Additionally, we show that increased biting behavior in association with this infertility in <i>Wolbachia</i>-infected mosquitoes may drive greater vector competence. These relationships require further exploration, given their ability to affect the success of field releases of <i>Wolbachia</i> for human disease reduction in drier climates where longer egg quiescence periods are expected.