An efficient and convenient method for collecting mosquito salivary proteins with high bioactivity and mosquito salivary viruses with high infectivity

Mosquito salivary proteins (MSPs) play vital roles in facilitating mosquito feeding and mediating mosquito-host interactions. The current standard method for collecting MSPs and mosquito salivary viruses (MSVs) is the forced salivation (FS) method, which is laborious and time-consuming, and yields MSPs with altered composition and reduced bioactivity and MSVs with weakened infectivity, compared to those secreted under physiological conditions. To address these shortcomings, we developed a collecting-by-natural-feeding (CNF) method which empowers convenient and time-saving collection of MSPs and MSVs under physiological conditions. Compared to the FS method, the CNF method enables collection of relatively large quantities of MSPs in a short period of time without any harm to the mosquitoes, allowing for dynamic collection from the same individuals. More importantly, the MSPs and the MSVs collected by the CNF method exhibited a drastically higher bioactivity or infectivity than those collected by the FS method. The CNF method requires no specialized materials or instruments, thus enhancing its general applicability. Collecting sufficient quantities of MSPs with high bioactivity and MSVs with high infectivity has long been a significant challenge. Our novel CNF method may overcome this obstacle, making it highly useful for research on MSPs and vector biology.