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Appl Environ Microbiol. 2019 Jul 26. pii: AEM.01202-19. doi: 10.1128/AEM.01202-19. [Epub ahead of print]

Adaptation of a microbial detection array as a monitoring tool revealed the presence of mosquito-borne viruses and insect-specific viruses in field-collected mosquitoes.

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Department of Entomology, Texas A&M University, College Station, Texas
Physical & Life Sciences Directorate, Lawrence Livermore National Laboratory, Livermore, California.
Department of Entomology, Texas A&M University, College Station, Texas.
Program of Biology, Texas A&M University-San Antonio, San Antonio, Texas.
Physical & Life Sciences Directorate, Lawrence Livermore National Laboratory, Livermore, California


Several mosquito-borne diseases affecting humans are emerging or re-emerging in the United States. The early detection of pathogens in mosquito populations is essential to prevent and control the spread of these diseases. In this study, we tested the potential applicability of the Lawrence Livermore Microbial Detection Array (LLMDA) to enhance bio-surveillance by detecting microbes present in Aedes aegypti, Aedes albopictus and Culex mosquitoes that are major vector species globally, including in Texas. The sensitivity and reproducibility of the LLMDA was tested in mosquito samples spiked with different concentrations of dengue virus (DENV) revealing a detection limit of >100 but <1000 pfu/mL. Additionally, field-collected mosquitoes from Chicago, Illinois and College Station, Texas of known infection status (West Nile virus (WNV) and Culex flavivirus (CxFLAV) positive) were tested on the LLMDA to confirm its efficiency. Mosquito field samples of unknown infection status, collected in San Antonio, TX and the Lower Rio Grande Valley (LRGV), TX were run on the LLMDA and further confirmed by PCR or qPCR. The analysis of the field samples with the LLMDA revealed the presence of cell fusing agent virus (CFAV) in Ae. aegypti populations. Wolbachia was also detected in several of the field samples (Ae. albopictus and Culex spp.) by the LLMDA. Our findings demonstrated that the LLMDA can be used to detect multiple arboviruses of public health importance including viruses that belong to the Flavivirus, Alphavirus and Orthobunyavirus genera. Additionally, insect-specific viruses and bacteria were also detected from field-collected mosquitoes. Another strength of this array is its ability to detect multiple viruses in the same mosquito pool allowing for the detection of co-circulating pathogens in an area, and the identification of potential ecological associations between different viruses. This array can aid in the bio-surveillance of mosquito borne viruses circulating in specific geographical areas.ImportanceViruses associated with mosquitoes have made a large impact on public and veterinary health. In the US, several viruses including WNV, DENV and chikungunya virus (CHIKV) are responsible for human disease. From 2015-2018, imported Zika cases were reported in the US and in 2016-2017, local Zika transmission occurred in the states of Texas and Florida. With globalization and a changing climate, the frequency of outbreaks linked to arboviruses will increase, revealing a need to better detect viruses in vector populations. With its capacity to detect viruses, bacteria and fungi, this study highlights the ability of the LLMDA to broadly screen field-collected mosquitoes and contribute to the surveillance and management of arboviral diseases.


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