Infecting and mosquitoes with Dirofilaria immitis and Brugia malayi v3

This protocol is for infecting Aedes aegypti and Aedes albopictus mosquitoes with the filarial nematodes Dirofilaria immitis or Brugia malayi. Although it is not necessary, it is strongly recommended to measure the number of ingested parasites, which indicates the exposure level for the mosquito and allows for better comparison between replicates. We use a post-feed treatment protocol, but if mosquito groups are treated prior to infection, then it is not just strongly recommended, but critical to measure ingested microfilariae in a group of mosquitoes immediately following the feed to confirm that treatment does not change the extent of blood feeding. Note that microfilariae are large (approximately 300 µm) and will quickly settle in blood samples. This should be considered throughout the protocol and this fact is pointed out in places in this protocol.