Isoenzyme variation in Aedes aegypti correlated with Dirofilaria immitis infectability

From the Vero Beach strain of the mosquito Aedes (Stegomyia) aegypti (L.) (Diptera: Culicidae), substrains were selected for susceptibility (SS) and refractoriness (RR) to the dog heartworm Dirofilaria immitis (Leidy) (Filarioidea: Onchocercidae). These two lines and their reciprocal F1 hybrids were analysed for genetic variation at 14 enzyme loci, using polyacrylamide gel electrophoresis. Six of the enzyme loci showed variation (sample size 48 alleles/locus/line). Three of these were monomorphic in the refractory line but polymorphic in the susceptible, i.e. aconitase hydratase (Acoh), isocitrate dehydrogenase-1 (Idh-1) and phosphoglucomutase (Pgm). The other three loci, glucose-6-phosphate isomerase (Gpi), hexokinase-1 (Hk-1) and isocitrate dehydrogenase-2 (Idh-2), were polymorphic in both SS and RR lines and their hybrids. At two loci (Hk-1, Pgm) three alleles were detected, whereas the other polymorphic loci had only two alleles. For Hk-1, the most frequent allele was Hk-1(80) (0.563) in refractory and Hk-1(100) in the susceptible (0.521) and F1 hybrids. For Pgm the most frequent alleles were Pgm125 in the susceptible line (0.646) and Pgm100 in the F1 hybrids (0.563 and 0.604) and refractory line (1.000). The mean observed heterozygosity (Ho), the mean Hardy-Weinberg expected heterozygosity (He) and the mean number of alleles per locus in the refractory line were lower, but not significantly so, than in the susceptible line and their reciprocal F1 hybrids; the proportion of polymorphic loci was significantly lower in the refractory than in the susceptible line and their F1 hybrids. Within both lines all polymorphisms were in Hardy-Weinberg equilibrium, whereas significant departures from predicted frequencies were observed in SS x RR hybrids at four polymorphic loci (Acoh, Gpi, Hk-1, Pgm) and at three polymorphic loci (Acoh, Hk-1, Pgm) in RR x SS hybrids. The average Nei's and modified Rogers' genetic distances between the lines were 0.024 and 0.139, respectively. These electrophoretic data show that the refractory line (putatively lacking fi allele) can be distinguished from the susceptible line (fi/fi) and their hybrids (heterozygous fi) by isozyme marker frequencies, but it remains to be seen whether this difference is causal or chance linkage. In any case, this model system of Ae. aegypti/D. immitis provides opportunities to better understand and manipulate the molecular biology of filariasis transmission.