Methods in Anopheles Research - MR4

Chapter 3 : Specific Anopheles Techniques3.7 Anopheles Mating3.7.3 Pair MatingsPage 1 of 23.7.3 Pair MatingsMR4 StaffBackgroundPair mating (or single-pair mating) is most commonly used when knowledge of the genetic makeup ofboth parents is important. The isolation of specific phenotypes within Anopheles mosquitoes is useful indetermining vector population species/genetic composition (Rabbani et al. 1976). Phenotypes such asinsecticide resistance as well as other novel genetic mutations can be quickly isolated and purified usingpair mating (Collins et al. 1986).Beside forced copulation (Chapter 3.7.2), mating between a particular male and female can be obtainedby free-mating in small numbers in small containers. Allowing a male and a female to mate freely is lessinvasive and time consuming than the method of forced copulation. Neither method is efficient, however.Benedict and Rafferty (2002) reported a method for obtaining reasonable frequencies of free matings.They observed that mating did not occur until later than is typical, so we recommend keeping the pair inthe mating tube for 7 or 8 days.Materials• Qorpack tubes with modified lids, or similar• Cotton balls• 10% sucrose solution• Rack for holding the single pair mating tubes• AspiratorProcedure1. Ensure the lighting in the space in which the mosquitoes will beheld is adjusted to obtain sunset and sunrise periods to entrain thelarvae before adulthood.2. After emergence, place a male and female in an individual rearingcontainer (Figure 3.7.3.1) and leave for 5-8 days. Additionalfemales may be added if desired – 5 would be a reasonablemaximum.3. Maintain the adults by placing a sugar pad on top of the tube andkeeping it wet.4. Obtain eggs as described in Chapter 3.9 - Family Culture.Figure 3.7.3.1. This particularchoice of tube that we use forfamily oviposition is modifiedfrom a chamber purchasedthrough Qorpack (Bridgeville,PA. No. 3891). Many similartubes are suitable. The bottommay be covered with a filterpaper disk to absorb fluids.ReferencesBenedict MQ, Rafferty CS (2002) Unassisted isolated-pair mating of Anopheles gambiae (Diptera:Culicidae) mosquitoes. Journal of Medical Entomology 39:942-944Collins FH et al. (1986) Genetic selection of a Plasmodium-refractory strain of the malaria vectorAnopheles gambiae. Science 234:607-610Rabbani MG, Seawright JA, Leatherwood LB (1976) A method for culturing single families of Anophelesalbimanus. Mosq. News 36:100-102