Malaria diagnosis Giemsa stained blood smear traditional method may be hard to detect low level infection hard to identify which Plasmodium species occurs 2 Immunofluorescence assay IFA antibody against specific malaria species conjugated to fluorophore observe with fluorescence microscope usually not available in hospital setting in developing countries where malaria is endemic 3 Molecular diagnostic tools to distinguish 4 malaria species eg PCR Ingested pathogens having already survived challenges from the vertebrate immune system must then enter the mosquito midgut and overcome The vertebrate immune system in ingested blood Digestive enzymes of mosquito Heme i e oxidative stress from the blood meal Endogenous microorganisms Peritrophic matrix of mosquito Mosquito immune responses 68 of An gambiae mosquitoes fed on blood containing P falciparum never became infected i e gametocytes in gut Overall 75 of infected mosquitoes never became infective i e sporozoites in salivary glands Mosquitoes have a highly effective innate immune system Insects don t have adaptive acquired immunity so no immunoglobulins Insects have only innate immunity the more evolutionarily ancient form Insect innate immunity includes both cellular and humoral defense responses a somewhat artificial distinction since many of the pathways and molecules are common to both systems Cellular defenses Hemocyte mediated Phagocytosis encapsulation Mosquitoes have three types of hemocytes granulocytes oenocytoids Prohemocytes Humoral defenses Toll IMD JAK STAT and PPO pathways PRRs kick off a signaling cascade Production of immune effectors antimicrobial peptides thioester containing proteins ROS NOS Entomological considerations The frequency of disease will depend on the Entomological Inoculation Rate Entomological Inoculation Rate number of infectious bites day EIR Product of HBR PIM HBR human biting rate bites by vectors person day PIM proportion of mosquitoes that are infectious e g for malaria PIM sporozoite rate The HBR will be influenced by the behavior of vectors Since most anopheles bite after dark when people are inside and asleep the tendency to follow an odor plume into houses is important Endophilic will enter houses to feed on hosts a big problem Exophilic bites outdoors only not as dangerous overall