Methods in Anopheles Research - MR4

Chapter 2 : Anopheles Laboratory Biology and Culture2.4 Anopheles Culture2.4.10 Bloodfeeding : Membrane Apparatuses and AnimalsPage 5 of 6Figure 2.4.10.5. The components that make up themembrane feeder. Clockwise from top, a membrane(in this case Parafilm; the manufacturer’smembranes are recommended), a completelyassembled membrane unit ready to be attached to theheating unit, the metal membrane unit without amembrane, and the “O” ring used to attach themembrane.Switching from a live animal to a membrane system can be difficult if a colony is accustomed to feedingon a live animal. There are some methods commonly employed to increase success rates when using amembrane feeder.Starvation: sugar-starving females 5-12 hours can be helpful in increasing desire to take a blood meal;sugar-starving can cause mortality in males and should be limited if the males are not first removed.Lighting: most successful membrane feeds are conducted in the dark; you can either cover the cagesbeing fed or turn off the lights.Stimuli: gently rubbing the membrane on your arm will transfer human volatiles needed for phagostimulationin mosquitoes.ATP: adding this chemical to the blood has been shown to improve membrane feeding with some strainsof Aedes but not Anopheles.Blood sourcesA reliable source of blood for membrane feeding must be found. Although blood can be drawn from ahuman volunteer or from a laboratory animal, the use of either type of blood may require specialpermission through an IRB or an IACUC (Institutional Animal Care and Use Committee). This typicallyinvolves a review as well as a trained specialist to perform the venipuncture procedure.Blood can sometimes be obtained from a slaughter house (an abattoir). If properly treated (eithercollected in a heparinized vial or provided some anti-coagulant agent and stored at 4°C), these sources ofblood have been used successfully for up to 2 months. Heparinized or citrinated animal blood can also beprocured from specialized vendors such as Lampire, HemoStat, and Bioreclamation.Personal safety while using a membrane feeding apparatus is critical since all blood should be handledas if it were infected. Proper training in the use of Personal Protective Equipment (PPE) such as latexgloves, lab coat, and face-shield should be provided to all personnel handling the blood or performing thefeedings. Personnel should receive appropriate instructions according to their institutions rules.ReferencesCosgrove JB, Wood RJ, Petric D, Evans DT, Abbott RH (1994) A convenient mosquito membrane feedingsystem. J Am Mosq Control Assoc 10:434-436