The Effect of SkitoSnack, an Artificial Blood Meal Replacement, on Aedes aegypti Life History Traits and Gut Microbiota

Kristina K Gonzales,Margaret Kowalski,Wayne A Van Voorhies,Immo A Hansen,Yiyi Li,Yashoda Kandel,Hae-Na Chung,F Omar Holguin,Kathryn A Hanley,Emily L Moore,Stephanie M Willette,Julia Vulcan,Stacy D Rodriguez

doi:10.1038/s41598-018-29415-5

Kristina K Gonzales, Margaret Kowalski + Show 11 more

Open Access

https://doi.org/10.1038/s41598-018-29415-5

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Journal: Scientific Reports	Publication Date: Jul 23, 2018
Citations: 31	License type: open-access

Affiliation: New Mexico State University

Abstract

Public health research and vector control frequently require the rearing of large numbers of vector mosquitoes. All target vector mosquito species are anautogenous, meaning that females require vertebrate blood for egg production. Vertebrate blood, however, is costly, with a short shelf life. To overcome these constraints, we have developed SkitoSnack, an artificial blood meal replacement for the mosquito Aedes aegypti, the vector of dengue, Zika and chikungunya virus. SkitoSnack contains bovine serum albumin and hemoglobin as protein source as well as egg yolk and a bicarbonate buffer. SkitoSnack-raised females had comparable life history traits as blood-raised females. Mosquitoes reared from SkitoSnack-fed females had similar levels of infection and dissemination when orally challenged with dengue virus type 2 (DENV-2) and significantly lower infection with DENV-4. When SkitoSnack was used as a vehicle for DENV-2 delivery, blood-raised and SkitoSnack-raised females were equally susceptible. The midgut microbiota differed significantly between mosquitoes fed on SkitoSnack and mosquitoes fed on blood. By rearing 20 generations of Aedes exclusively on SkitoSnack, we have proven that this artificial diet can replace blood in mosquito mass rearing.

Highlights

Aedine mosquito species transmit viruses which cause diseases such as yellow fever, dengue fever, chikungunya fever, and Zika congenital syndrome[1,2]
We performed an extensive series of experiments to optimize the mixture of nutrients for SkitoSnack for Ae. aegypti females in order to maximize egg numbers and hatch rates
We found no significant difference between the body infection rates (Fisher’s exact test, df = 1, p = 1.0, N = 115), mean virus titer from infected bodies (t-test, t = −0.288, df = 17, p = 0.78), dissemination rates (Fisher’s exact test, df = 1, p = 0.58, N = 19), and mean virus titer from infected heads (t-test, t = 0.0126, df = 14, p = 0.99) of females raised on bovine blood or SkitoSnack challenged with a dengue virus type 2 (DENV-2)-infected rabbit blood meal

Summary

Introduction

Aedine mosquito species transmit viruses which cause diseases such as yellow fever, dengue fever, chikungunya fever, and Zika congenital syndrome[1,2]. Progress in gene editing techniques, in particular CRISPR/Cas 9, has opened up new avenues for the usage of gene-drive systems in SIT18–20 Another promising approach to control mosquito-borne disease is the release of mosquitoes infected with the bacterial endosymbiont Wolbachia. When mosquitoes are to be used for studies of vector competence, the impact of rearing on the artificial diet on vector competence must be defined, and the vehicle should only be used for pathogen delivery to vectors if subsequent levels of infection are similar to those seen when the pathogen is delivered in blood We have used these requirements as a foundation for the formulation and testing of our artificial blood meal, SkitoSnack. We report differences in the midgut microbiota between blood-fed and SkitoSnack-fed mosquitoes and demonstrate that SkitoSnack is an appropriate vehicle for dengue virus delivery

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