Background
Panstrongylus megistus is the most important vector of Chagas disease in Brazil. Studies show that the principal factor hindering the control of triatomines is reinfestation of houses previously treated with insecticides. Studies at the microgeographic level are therefore necessary to better understand these events. However, an efficient molecular marker is not yet available for carrying out such analyses in this species. The aim of the present study was to identify and characterize microsatellite loci for future population genetic studies of P. megistus.
Methods
This study work consisted of five stages: (i) sequencing of genomic DNA; (ii) assembly and selection of contigs containing microsatellites; (iii) validation of amplification and evaluation of polymorphic loci; (iv) standardization of the polymorphic loci; and (v) verification of cross-amplification with other triatomine species.
Results
Sequencing of males and females generated 7,908,463 contigs with a total length of 2,043,422,613 bp. A total of 2,043,690 regions with microsatellites in 1,441,091 contigs were obtained, with mononucleotide repeats being the most abundant class. From a panel of 96 loci it was possible to visualize polymorphisms in 64.55% of the loci. Of the 20 loci genotyped, the number of alleles varied from two to nine with an average of 4.9. Cross-amplification with other species of triatomines was observed in 13 of the loci.
Conclusions
Due to the high number of alleles encountered, polymorphism and the capacity to amplify from geographically distant populations, the microsatellites described here show promise for utilization in population genetic studies of P. megistus.
Graphic abstract
Background
Triatoma brasiliensis
Neiva, 1911 is the main vector of
Trypanosoma cruzi
in the caatinga of Northeastern Brazil. Despite of its epidemiological relevance, there are few studies on its genetic variability. Using microsatellite markers, we characterized the variability and dynamics of infestation and reinfestation of
T. brasiliensis
after residual insecticide spraying in five surveys conducted in a well-defined rural area located in the municipality of Tauá, Ceará, between 2009 and 2015. We evaluated: (1) general variability among local of captures; (2) variability along the time analysis (2009, 2010 and 2015); (3) and reinfestation process.
Results
On the analysis (1) global and pairwise
F
ST
values suggested absence of clusters among the area. AMOVA indicated that total variation is mainly represented by individual differences. Absence of clustering indicates a panmitic unit, with free gene flow. For (2), Pairwise
F
ST
indicated alterations in the genetic profile of the triatomines along the time. (3) Analysis of the reinfestation process showed that the domiciliary units investigated had different sources of infestation despite of its proximity.
Conclusions
Observed homogeneity can be explained by the great dispersal capacity of
T. brasiliensis
, overlapping the different environments. Persistent house infestation in Tauá may be attributed to the occurrence of postspraying residual foci and the invasion of triatomines from their natural habitats.
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