Methods in Anopheles Research - MR4

Chapter 2 : Anopheles Laboratory Biology and Culture2.4 Anopheles Culture2.4.10 Bloodfeeding : Membrane Apparatuses and AnimalsPage 3 of 6Figure 2.4.10.1. Use of arabbit for mosquito feedingutilizing a combination ofdrugs for anesthesia andpain under an IRBapprovedprotocol.Restraining the animalensures uninterruptedfeeding for the mosquitoesand safety for the rabbitshould it awakenunexpectedly. Shaving thebelly prior to feedingmakes the skin easilyaccessible.Membrane feedingDue in part to the restrictions on the use and difficulty of live animals in a research setting, artificialmembrane methods for feeding have been developed. Designs all have two basic features: a heatingelement and a membrane to hold the blood. The heating element is necessary to maintain the blood at areasonable temperature at which mosquitoes will imbibe, typically between 35- 40°C. The membraneshould be relatively thin so mosquitoes can pierce it easily but sturdy so that spilling does not occur.Membrane typesThere are several materials that have been used in membrane feeding. Natural membranes are claimedto work best (e.g. animal skin membranes), but they are can be difficult to procure and can usually onlybe used once (Novak et al. 1991). Other membranes used with some success include: Parafilm M®,collagen sheets, latex membranes e.g. gloves or dental dam, sausage casings (natural or synthetic),Baudruche membranes (Joseph Long Inc, N.J.), and condoms (lambskin or latex).Membrane FeedersSeveral different methods have been developed for feeding mosquitoes, some simple, some are morecomplex. A short list of some reported examples from the literature are:Latex or lambskin condoms: Fill the condom with heparinized blood. Tie it closed and soak it in a warmwater bath (~40°C) for 30 minutes to 1 hour to fully heat the blood. Condoms have been reported to havevarying levels of success.Tseng feeder (Tseng 2003): This employs a Parafilm packet filled with blood then wrapped in a wire meshcasing. It is best to heat the blood before pouring it into the Parafilm packet.Mishra feeder (Mishra et al. 2005): A Petri dish is wrapped with Parafilm on the outside leaving a smallpocket in the center of the bottom. Warm water is poured into the Petri dish and a small amount of bloodis injected into the Parafilm. This unit can be placed directly on the cage.Mourya feeder (Mourya et al. 2000): A 10 mm hole is drilled into a sheet of acrylic. Parafilm is thenstretched over the sheet and warmed blood added into the 10 mm hole. A beaker of warm water placedon the top of the hole maintains the temperature.More Complex Designs for Membrane FeedersGlass feeders: Although there are several models of these, most are built in the same manner: An outerarea contains circulating warm water and an inner chamber where the blood is poured, see Figure2.4.10.2. A nice reference to several styles is given by Kasap et al. (2003).