An isoenzymatic comparative analysis of the variability and genetics differentiation among

RESUMO -Modificações na expressão gênica foram observadas nos sistemas esterase, leucina aminopeptidase e x-glicerofosfato desidrogenase, durante o desenvolvimento ontogenético de Anopheles albitarsis. A esterase revelou quatro regiões de atividade, sendo a esterase 1 detectada apenas em larvas de 4º estádio velhas e em pupas, as esterases 2 e 4 foram presentes durante todo o desenvolvimento, e a esterase 3 revelou-se praticamente apenas em larvas e raríssimas vezes em pupas. Também foram observadas quatro regiões de atividade na leucina am presente somente em pupas e adultos e a LAP4 foi detectada nos três diferentes estágios. Uma única região foi observada para a x-glicerofosfato desidrogenase e a intensidade de sua atividade cresce à medida que se aproxima o estágio adulto. Palavras-chave:Anopheles albitarsis, isoenzimas, ontogenia, malária, Amazônia. Ontogenetic Patterns of Esterases, Leucine Aminopeptidase and x-Glycerophosphate DehydrogenABSTRACT -Modifications in genetic expression were observed in the esterase, leucine albitarsis. The esterase revealed four regions of activity. Esterase 1 was detected in 4th stage larvae and in

Eighteen enzymatic loci were analysed in Aedes aegypti populations from four neighbourhoods in the city of Manaus. The analyses showed that the Downtown population was the most polymorphic (p = 55.6%) with higher observed and expected mean heterozygosities (H o = 0.152 ± 0.052; H e = 0.174 ± 0.052, respectively). The least variability was detected in the Coroado and Cidade Nova populations, both with polymorphism of 44.4%. The latter population presented the least observed heterozygosity (H o = 0.109 ± 0.037). Wright's F statistics showed that the mean value of F is was higher than that of F st (F is = 0.164 > F st = 0.048), and from analysis of molecular variance (AMOVA) it was found that 95.12% of the variability is found within populations indicating a certain intra-population differentiation possibly of the microgeographic structure resulting from some barrier in the random coupling. Although the four populations were similar genetically (D = 0.003 to 0.016), the 4.88% differentiation was significant.

Four populations of Aedes aegypti from Manaus were studied, using allozymes and RAPD loci, to determine intra-and interpopulation genetic variability and differentiation and to compare genetic structure parameters assessed with both markers. Five RAPD primers produced 52 polymorphic fragments, whereas only seven of 18 isozyme loci were polymorphic. The population from Praça 14 was the most polymorphic (P= 94.23% and P= 55.6%); while those from Coroado (P= 82.69% and P= 44.40%) and from Cidade Nova (P= 84.61% and P= 44.40%) were the least polymorphic, for both RAPD and isozymes respectively. The observed heterozygosity was higher between populations (Ho= 0.33-0.38) as assessed by RAPD. Wright's F statistics showed an F is value higher than F st (F is = 0.164 > F st = 0.048). AMOVA indicated that 95.12% of the genetic variability is intrapopulational. Even so, both of the genetic markers evaluated showed a relatively high gene flow ((N m = 15.15), and possibly are still random couplings, although the F is value was not low. The genetic distance between populations was similarly low for both markers: RAPD (0.012-0.016) and Isozymes (0.003-0.016). These results show that as assessed by both markers, the populations are genetically similar, and that isozymes (codominant) are the most efficient to detect the population genetic structure. Although isozymes revealed less genetic diversity than RAPDs, the estimated levels of genetic distance were identical.

Populations of Anopheles triannulatus from Macapá (AP), Aripuanã (MT), Ji-Paraná (RO), and Manaus-Janauari Lake (AM) were studied using 16 enzymatic loci. The results of the isozyme analysis showed that the population of Macapá presented higher polymorphism (56.3%). The lowest variability was observed in the population of Manaus (p = 25.0; Ho = 0.077 +/- 0.046). The results of Wright's F statistics showed unbalance due to excess of homozygotes (F(is) > F(st)), denoting a certain intrapopulational differentiation. Although the populations are genetically very close (D = 0.003-0.052), the dendrogram separates the populations in two groups: Macapá separated from that of Manaus, Ji-Paraná, and Aripuanã. This result may suggest a reduction in the genic flow, which possibly had some influence in the substructuration of the populations.

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Introduction: Isoenzymatic analyses were performed involving species of the Nyssorhynchus and Anopheles subgenera in order to estimate the intra and interspecies genetic variability. Methods: Mosquitoes were caught at different localities in the Amazon region. The collection and rearing of mosquitoes in the laboratory followed specific protocols. For the genetic variability analyses, the technique of horizontal electrophoresis on starch and starch-agarose gel with appropriate buffer systems was used. The alloenzyme variation was estimated using the Biosys-1 software. Results: Out of the 13 loci, eight were polymorphic. Anopheles nuneztovari presented the largest number of alleles per locus, while the smallest number was detected in Anopheles marajoara from Macapá. The largest number of polymorphic loci was found for Anopheles marajoara from Maruanum and the smallest for Anopheles benarrochi (Guayará Mirim). Anopheles darlingi (Macapá) presented the greatest heterozygosity (Ho = 0.167 ± 0.071), while the lowest heterozygosity (Ho = 0.045 ± 0.019) was observed in Anopheles intermedius (Pacoval) of the subgenus Anopheles. Wright's F coefficient revealed considerable genetic structuring between the populations of Anopheles darlingi (Fst = 0.110) and between the populations of Anopheles marajoara (Fst = 0.082). Conclusions: Considering all the species studied, the genetic distance ranged from 0.008 to 1.114. The greatest distance was between Anopheles mattogrossensis and Anopheles oswaldoi, while the smallest was between the Anopheles benarrochi populations.

Changes in the expression of genes were observed during development in populations of Anopheles (Anopheles) intermedius and Anopheles (Anopheles) mattogrossensis. Esterase showed seven zones of activity: EST1 was present in all developmental stages of both species; EST2 was observed only in larvae of A. intermedius and larvae and pupae of A. mattogrossensis, with greater activity in pupae; EST3 and EST5 were present in all developmental stages, with greater intensity in larvae; EST4 and EST6 showed weak activity in larvae of A. mattogrossensis and was not found in A. intermedius. Leucine aminopeptidase showed four zones of activity, of which LAP1 and LAP2 were found in all stages of A. intermedius, with highest activity in larvae, and in larvae only of A. mattogrossensis. LAP3 was detected in all stages of A. mattogrossensis and in larvae only of A. intermedius. LAP4 was detected only in larvae and pupae of A. mattogrossensis, with greater intensity in pupae. alpha-Glycerophosphate dehydrogenase showed a single zone of activity, detected in older fourth-instar larvae and becoming more intense from the pupal stage onwards.