Female A. aegypti that are homozygous for pls, a recessive gene on linkage group 2, cannot develop oocysts of P. gallinaceum. The gene has not been found in strains from Asia or the Americas but was present in all of 8 strains tested from Africa (Ghana, Nigeria, Tanzania, Uganda). Estimated gene frequency ranged from 0.27 to 1.0. Experimental laboratory populations were initiated with different proportions of pls, allowed to breed continuously for 8 months, and sampled periodically. Homozygous populations (all or no pls) remained unchanged; mixed populations showed no strong selection against pls. Most laboratory strains of Aedes aegypti (L.) are uniformly susceptible to Plasmodium gallinaceum Brumpt. Most parasitologists consider this susceptibility as a species characteristic. However, Kilama (1969, 1970) and Kilama and Craig (1969) showed that mosquitoes that are homozygous for the gene pls (plasmodium-susceptibility) cannot develop oocysts of P. gallinaceum. In refractory mosquitoes, the ookinete moves through the gut wall but the newly formed oocyst dies within 40 to 60 hr of the infectious blood meal. The gene pls behaves as a simple Mendelian autosomal recessive factor, located on linkage group 2. The pls factor was originally isolated from two strains of A. aegypti formosus (Walker) from West Africa (Kilama and Craig, 1969). Tests on more than 20 laboratory strains which had originated from Asia or the Americas showed uniform susceptibility; there was no evidence of pls in these parts of the world. These findings stimulated other questions: (1) Is pls restricted to West Africa? (2) Alternatively, could pls be found throughout tropical Africa, associated with the nondomestic subspecies, A. aegypti formosus? (3) What is the frequency of pls in field populations? (4) What is the selective value of pls and can this be tested in artificial populations in the laboratory? The present work reports experiments attempting to answer the above questions. The work was performed in 1969-70. Received for publication 11 November 1971. * Supported in part by U. S. NIH Research Grant No. AI-02753. t Present address: Faculty of Medicine, University of Dar es Salaam, Tanzania. MATERIALS AND METHODS The 8A strain of P. gallinaceum was used in these studies. Methods for maintaining malaria in chickens and for rearing and infecting mosquitoes were those reported by Kilama and Craig (1969). The strains of A. aegypti used are listed in Table I. All strains are currently available at the WHO International Reference Centre for Aedes at the University of Notre Dame. Individual mosquitoes may be refractory to malaria for reasons other than the presence of pls. Hence, the presence and frequency of pls was indirectly determined by using a complementary test; i.e., 2 recessive mutations in a transheterozygote are usually allelic if they produce a mutant phenotype (refractory) and nonallelic if they give wild-type progeny (susceptible). Individuals from the strain to be tested were therefore crossed with individuals from the ILOBI line which was homozygous for pls and therefore refractory. The F1 progeny from single-pair crosses were tested and segregated for susceptibility to P. gallinaceum. Experimental populations were established in the laboratory to follow the frequency of pls over a period of 8 months. Five populations were initiated, with the following proportions of pls to wild-type individuals: 10:0, 9:1, 1:1, 1:9, 0:10. Cages were 61 by 61 by 61 cm. The initial population in each cage consisted of 500 males and 500 females in the proper proportion. The mosquito populations were blood-fed twice per week on anesthetized mice. Each population received a constant supply of apple slices or sugar cubes. A large dishpan containing water and measuring 31 by 39 by 17 cm supplied an oviposition site. The water level in the pans was periodically raised to stimulate hatching. The larvae were fed on a solution of liver powder (Nutritional Biochemical Corporation). The frequency of pls in each population was determined monthly. Eggs were removed from each cage, conditioned, and hatched. The resulting adult females when approximately a week old were fed on an infected chicken, and scored for susceptibility a week later. The number of oocysts per mosquito stomach was counted; mosquitoes showing 0 to 10 oocysts were considered as re-
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