A Method for Repeated, Longitudinal Sampling of Individual Aedes aegypti for Transmission Potential of Arboviruses.

E Handly Mayton,Heather M Hernandez,Christopher J Vitek,Rebecca C Christofferson

doi:10.3390/insects12040292

E Handly Mayton, Heather M Hernandez + Show 2 more

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https://doi.org/10.3390/insects12040292

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Abstract

Mosquito-borne viruses are the cause of significant morbidity and mortality worldwide, especially in low- and middle-income countries. Assessing risk for viral transmission often involves characterization of the vector competence of vector-virus pairings. The most common determination of vector competence uses discreet, terminal time points, which cannot be used to investigate variation in transmission aspects, such as biting behavior, over time. Here, we present a novel method to longitudinally measure individual biting behavior and Zika virus (ZIKV) transmission. Individual mosquitoes were exposed to ZIKV, and from 9 to 24 days post-exposure, individuals were each offered a 180 μL bloodmeal every other day. Biting behavior was observed and characterized as either active probing, feeding, or no bite. The bloodmeal was then collected, spun down, serum collected, and tested for ZIKV RNA via qRT-PCR to determine individuals' vector competence over time. This included whether transmission to the bloodmeal was successful and the titer of expectorated virus. Additionally, serum was inoculated onto Vero cells in order to determine infectiousness of positive recovered sera. Results demonstrate heterogeneity in not only biting patterns but expectorated viral titers among individual mosquitoes over time. These findings demonstrate that the act of transmission is a complex process governed by mosquito behavior and mosquito-virus interaction, and herein we offer a method to investigate this phenomenon.

Highlights

Vector-borne viruses remain a major cause of morbidity in low- and middle-income countries and have been making incursions into more temperate regions recently
Cells were inoculated onto Vero cells at a multiplicity of infection (MOI) of 1
Todetermined determine by thepreviously limit of detection our method, we first measured the recovera24 dpe, observedofvector competence studies and average time to bility of virus from sera using a controlled scenario

Summary

Introduction

Vector-borne viruses remain a major cause of morbidity in low- and middle-income countries and have been making incursions into more temperate regions recently [1,2,3]. Dengue virus (DENV), chikungunya virus (CHIKV), and Zika virus (ZIKV) are transmitted by Aedes aegypti, an urban-dwelling mosquito widespread throughout tropical and subtropical areas [4,5,6,7,8]. The viruses this vector species transmits are responsible for large outbreaks affecting millions of people every year [9,10,11,12,13]. Complete understanding of the transmission systems of these arboviruses provides insight into the spread of the virus, especially when parameterizing prediction models that may be used in decision-making [14,15].

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