ABSTRACT. The Anopheles albitasis complex includes 6 species, and 3 are considered as malaria vectors in Brazil. Twenty-five polymorphic microsatellite DNA loci were isolated and characterized in 24-36 individuals from the neighborhood of Puraquequara, Manaus, Amazonas State, Brazil. The number of estimated alleles ranged from 2 to 10, the observed heterozygosity ranged from 0.182 to 0.897, and the expected heterozygosity ranged from 0.260 to 0.854. Eleven loci showed significant deviation from Hardy-Weinberg equilibrium. Eleven loci were cross- amplified successfully in 5 Anopheles species. These microsatellite loci will be useful in studies investigating population structure and evolutionary genetics in A. albitarsis sensu lato and other A. albitarsis complex species.
The genome assembly of Anopheles darlingi consists of 2221 scaffolds (N50 = 115,072 bp) and has a size spanning 136.94 Mbp. This assembly represents one of the smallest genomes among Anopheles species. Anopheles darlingi genomic DNA fragments of ~37 Kb were cloned, end-sequenced, and used as probes for fluorescence in situ hybridization (FISH) with salivary gland polytene chromosomes. In total, we mapped nine DNA probes to scaffolds and autosomal arms. Comparative analysis of the An. darlingi scaffolds with homologous sequences of the Anopheles albimanus and Anopheles gambiae genomes identified chromosomal rearrangements among these species. Our results confirmed that physical mapping is a useful tool for anchoring genome assemblies to mosquito chromosomes.
The caparari (Pseudoplatystoma tigrinum) is a widely distributed species in the Amazon Basin. It has a high market value and it is commonly consumed by the local population. In order to protect the species from possible overfishing, there is a need for a better understanding of the population diversity, since knowledge regarding its biology is scarce, which makes it difficult to monitor. Seven microsatellite loci (simple sequence repeats-SSR) for the species were isolated and characterized in 46 individuals which were sampled in four locations from the Madeira River, in the Brazilian Amazon. The number of alleles per locus ranged from three to eleven. The observed and expected heterozygosity ranged from 0,326 to 0,705 and 0,322 to 0,758, respectively. No linkage disequilibrium between pairs of loci was detected. The seven microsatellite loci were additionally used for inter-specific amplification in other four species of Pseudoplatystoma. Therefore, this study contributes to the first molecular species-specific SSR markers, which can be used as a new tool for estimating the genetic variability of P. tigrinum and has potential for application in population-related studies.
Background Anopheles darlingi is a monotypic species in terms of its morphological, genetic, and behavioral aspects and is the primary transmitter of human malaria (99%) in Brazil, especially in the Brazilian Amazon. In this pioneering study, 15 expressed sequence tag (EST)-simple sequence repeat (SSR) markers were obtained and characterized in samples from the municipality of São Gabriel da Cachoeira, Amazonas state, Brazil, with polymorphisms that can be used for further genetic research.
Methods and Results The specimens (from egg to larval stage) collected were bred in the insectary at INPA (National Institute for Amazonian Research). The SSR repeats within the contigs of the A. darlingi EST banks were confirmed on the Vector Base site. DNA was extracted and amplified using polymerase chain reaction and then genotyped. Fifteen polymorphic SSR loci were identified and characterized. The number of alleles totaled 76, and ranged from 2 to 9. The observed heterozygosity varied between 0.026 and 0.769, the expected heterozygosity between 0.025 and 0.776, and the mean polymorphism information content was 0.468. Eight loci showed Hardy-Weinberg equilibrium (HWE) after Bonferroni correction (P: (5%) ≤ 0.0005). No linkage disequilibrium was found among the loci.
Conclusions The polymorphic SSRs of the loci have been shown to be efficient for investigation of the variability and genetic population structure of A. darlingi.
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