Anopheles (Kerteszia) cruzii infected by Plasmodium in the Atlantic Forest indicates that the malaria transmission cycle is maintained even after howler monkeys' population decline.

The Atlantic Forests outside of the Amazon region in Brazil are low-frequency malaria hotspots. The disease behaves as a zoonosis maintained by nonhuman primates (NHPs), especially howler monkeys. Between 2016 and 2018, Brazil witnessed the largest yellow fever outbreak since 1980, resulting in massive declines in these NHP populations. However, reports of malaria cases continued in transmission areas. This scenario motivated this survey to determine the frequency of infection of the anophelines by Plasmodium species. Mosquitoes were captured using Shannon traps and CDC light traps and identified as to species based on morphological characters. The screening for malaria parasites targeted only Anopheles species belonging to the subgenus Kerteszia, the proven primary malaria vector. A TaqMan qPCR assay using ribosomal primers (18S rRNA gene) was performed in a Step One Plus Real-time PCR to detect Plasmodium species. Seven hundred sixty field-caught anophelines divided into 76 pools were examined. Out of 76 tested pools, seven (9.21%) were positive. Three pools were Plasmodium malariae-positive, and four were Plasmodium vivax-positive. The anopheline infection was expressed as the maximum infection rate (MIR), disclosing a value of 0.92%, indicative of a steady state. Such stability after the yellow fever outbreak suggests that other species of NHPs could support transmission.