Stability and detection of nucleic acid from viruses and hosts in controlled mosquito blood feeds.

Ethan K Jackson,Coyne Drummond,Andrzej Pastusiak,Maria Teresa Sáenz Robles,Mary E Gebhardt,Douglas E Norris,Giovanna Carpi,Isaiah Hoyer,Michael R Reddy,James M Pipas,Luciano Andrade Moreira

doi:10.1371/journal.pone.0231061

Abstract

Monitoring the presence and spread of pathogens in the environment is of critical importance. Rapid detection of infectious disease outbreaks and prediction of their spread can facilitate early responses of health agencies and reduce the severity of outbreaks. Current sampling methods are sorely limited by available personnel and throughput. For instance, xenosurveillance utilizes captured arthropod vectors, such as mosquitoes, as sampling tools to access blood from a wide variety of vertebrate hosts. Next generation sequencing (NGS) of nucleic acid from individual blooded mosquitoes can be used to identify mosquito and host species, and microorganisms including pathogens circulating within either host. However, there are practical challenges to collecting and processing mosquitoes for xenosurveillance, such as the rapid metabolization or decay of microorganisms within the mosquito midgut. This particularly affects pathogens that do not replicate in mosquitoes, preventing their detection by NGS or other methods. Accordingly, we performed a series of experiments to establish the windows of detection for DNA or RNA from human blood and/or viruses present in mosquito blood meals. Our results will contribute to the development of xenosurveillance techniques with respect to optimal timing of sample collection and NGS processing and will also aid trap design by demonstrating the stabilizing effect of temperature control on viral genome recovery from blood-fed mosquitoes.

Highlights

Emerging and re-emerging viral infectious diseases represents a continued threat to human health and imposes an immense economic burden on at-risk populations [1,2,3,4,5]
Nucleic acid content of mosquitoes fed with virus-containing blood meals was assessed by qPCR
dengue virus 2 New Guinea strain C (DENV-2) is an arbovirus with tropism for Ae. aegypti, and was expected to be detectable by qPCR once the virus infected midgut tissues and started replicating with a seven to ten day extrinsic incubation period (EIP) [26]

Summary

Introduction

Emerging and re-emerging viral infectious diseases represents a continued threat to human health and imposes an immense economic burden on at-risk populations [1,2,3,4,5]. Monitoring for emerging infectious diseases remains a challenge, especially in under-developed parts of the world, due to inaccessibility of remote locations, difficulty linking zoonotic and human. Stability of nucleic acids in mosquito bloodmeals gift was intended to support in the form of salaries for JMP and DEN. The funder provided support in the form of salaries for authors [IH, AP, MRR, EKJ], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section

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Discussion

Conclusion