Adaptation of a microbial detection array as a monitoring tool revealed the presence 2 of mosquito-borne viruses and insect-specific viruses in field-collected mosquitoes

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 \u3e100 but \u3c1000 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