Despite widespread availability of long-lasting insecticide treated bednet (LLINs), malaria transmission with high human infection incidence continues in Malawi. We hypothesize that biases in mosquito human feeding could be driving infection transmission. The study was conducted to quantify the extent and diversity of human blood feeding by vector mosquitoes in areas with extensive use of LLINs including Balaka district with standard permethrin and Machinga district with piperonyl butoxide synergist LLINs (PBO). Mosquitoes were sampled indoors during 2019-2020 by pyrethrum spray catch and light traps. Human demographic data and blood spots on filter paper by finger prick were collected from consented participants. Mosquitoes were identified by morphological and molecular methods. Mosquito blood meal source and Plasmodium falciparum infection in abdomen or head-thorax of mosquitoes was determined by qPCR methods, using hostspecific oligonucleotide probes and parasite-specific probes, respectively. Human blood meals and human blood spots were analyzed by genotyping 24 human microsatellite loci, to generate genetic profiles that were matched using an algorithm executed in R package. Of 635 blood-fed Anopheles spp, 44.1% were An. arabiensis, 16.2% An. gambiae s.s, 33.5% An. funestus, 0.3% An. parensis, and 5.8% were unidentified Anopheles spp. Blood meals were predominantly from humans (80.5%), but also goats (5.2%), dogs (2.0%), human/goat mixed (11.1%), human/dog (0.9%), and dog/goat (0.2%). Samples from Balaka with standard LLIN had a higher human blood index (81.2%) than Machinga (75.3%) with PBO nets, but it was not significantly different (p = 0.59). Sporozoite infection in the head-thorax was 16% (99/633) overall, for an estimated annual EIR of 26 infectious bites/person. Genotyping analysis revealed a highly non-random pattern of human host section with a bias toward feeding upon males aged 6-15 years. These results show a high proportion of human blood meals, regardless of LLIN type, and suggest that the school age males may be important contributors to transmission.