Methods in Anopheles Research - MR4

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Chapter 2 : Anopheles Laboratory Biology and Culture2.1 Behavior and Physiology of Anophelines in the LaboratoryPage 5 of 8Figure 2.1.11 From left to right, Anopheles gambiae larvae 24 hours post hatch (1 st instar or L1), 2 dayspost hatch (2 nd instar or L2), 5 days post hatch (3 rd instar or L3) and 6 days post hatch (4 th instar or L4).All were photographed at the same magnification.Figure 2.1.12. Anopheles gambiae pupae: twofemales (bottom right) and one male (top leftcorner). Size difference is not obvious by eye.Figure 2.1.13. Aedes aegypti pupae: twomales (right) one female (left). Size disparitiesare apparent.
Chapter 2 : Anopheles Laboratory Biology and Culture2.1 Behavior and Physiology of Anophelines in the LaboratoryPage 6 of 8NutritionThe amount of available food significantly affects larval growth. Underfeeding can cause as much delayas overfeeding but will likely be evident later, especially in the adult stage. Much has been written aboutDietary Restriction (DR) in mice and flies and is reviewed in Chapter 3 for its contributions to longevityand fecundity. In short, DR causes the animals to live longer but age faster having negative influence onfecundity and tolerance for environmental fluctuations and infection.Larval DensityAchieving the right density is very important in growth and development. The most common problemsassociated with over crowding are: longer development time, reduced pupation and eclosion, and adecrease in pupal weight. See Chapter 2 Culture section for more on proper density for anophelines.Effect of Larval Health on AdultsAdults from larvae that were crowded are typically smaller and less fecund. The ultimate size of an adultmosquito will be based on genetics in combination with the environmental conditions experienced throughdevelopment. Studies have shown that larvae that are reared in crowded conditions had negative effectson weight at emergence, quantity of the blood meal and overall fertility. Poor larval conditions cannot betotally overcome by good diet or care in later stages, therefore careful attention to larval conditionsdetermines high overall quality of production.Environmental effects on rhythmsStudies show a link between environmental factors and ecdysis. These studies are limited to certainspecies and conditions; however, the evidence supports the shifting of ecdysis under temperaturechanges, light/dark cycles, and larval stress such as salinity. For example, researchers found that incontinual darkness, with variable temperature cycles, the larval-pupal ecdysis was more likely to occurduring the warm phase.The time of day/night the ecdysis will occur is species-dependent. It is thought that the trigger to molt isswitched on and off based on a daily rhythmic activity cycle of 24-hour intervals that is exhibited by manyorganisms, or a circadian rhythm. This is not universal among all mosquitoes. Examples of some foundnot to have such a rhythm are An. quadrimaculatus (Nayar and Sauerman 1970) and Ae. aegypti(Haddow et al. 1959). The best rule of thumb is if your insectary has problems with ecdysis beingtemporally irregular or extended over several days, experiment with your conditions to correct theproblem. Light and temperature are the key factors.Adult feedingPlant JuicesBoth adult male and female mosquitoes will drink plant juices as an energy source (see review in (Foster1995). Plant sugar is the major food resource for mosquitoes. In the wild, the most common source isfloral nectar, but other sources exist such as damaged fruit (Figure 2.1.14) or vegetative tissue. Withthese different meals, the mosquito would be receiving largely sucrose, fructose, or glucose, dependingon the source. Other sources such as maltose or melibiose are seldom found in mosquitoes. Amino acidsneeded for ovary development can be found in some nectar, but the concentrations are not high enoughto replace a blood meal. Natural sugar sources can have a wide range of amounts of sugar from none to50% w/v, though 20-50% w/v is the normal range. Mosquitoes have been seen ingesting crystallizedsucrose by liquefying it with saliva.
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Methods in Anopheles Research - MR4

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