Percorrer por autor "Balata, Duarte"
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- Exploring the aquatic resistomePublication . Rosado, Tânia; Manageiro, Vera; Balata, Duarte; Menezes, Carina; Ferreira, Eugénia; Paulo, Octávio; Caniça, Manuela; Dias, ElsaAntibiotic resistance is one of the major problems in public health today since the failure of antibiotherapy has dramatic clinical implications such as the increase of mobility/mortality and of health costs. Antibiotic resistance is a dynamic process since antibiotic-resistant microorganisms, and the corresponding genetic material, flows and disseminates among several settings simultaneously: humans, animals and natural habitats. Water environments are recognized as important pools of antibiotic pollution and antibiotic resistance genes, but the water resistome (collection of all genes responsible for antibiotic resistance in water environments) is far from being characterized. It is well known that antibiotic pollution affects the structure/functioning/diversity of aquatic ecosystems, namely the cyanobacteria community. However, the role of cyanobacteria in the context of antibiotic resistance was never characterized. In this presentation, the aims, the team, the methodologies and the preliminary results of the national project EXPLORAR – Exploring the Aquatic Resistome (PTDC/BIA-BMA/31451/2017), founded by the Portuguese Foundation for Science and Technology (FCT, PT) will be discussed. In this project, we are investigating the contribution of indigenous freshwater organisms (cyanobacteria and bacteria) to water resistome. We expect to identify and characterize ecological niches associated with antibiotic resistance in freshwater environments. This may contribute to define a monitoring strategy to map the antibiotic resistance profiles of national freshwater resources. We expect to contribute to the definition of preventive measures against the dissemination of antibiotic resistance in the environment.
- Prediction of Antibiotic Resistance Genes in Cyanobacterial Strains by Whole Genome SequencingPublication . Balata, Duarte; Rosado, Tânia; Pina-Martins, Francisco; Manageiro, Vera; Menezes, Carina; Ferreira, Eugénia; Paulo, Octávio S.; Caniça, Manuela; Dias, ElsaCyanobacteria are ubiquitous in freshwater environments, but their role in aquatic resistome remains unclear. In this work, we performed whole genome sequencing on 43 cyanobacterial strains isolated from Portuguese fresh/wastewaters. From 43 available non-axenic unicyanoabacterial cultures (containing only one cyanobacterial strain and their co-occurring bacteria), it was possible to recover 41 cyanobacterial genomes from the genomic assemblies using a genome binning software, 26 of which were classified as high-quality based on completeness, contamination, N50 and contig number thresholds. By using the comprehensive antibiotic resistance database (CARD) on the assembled samples, we detected four antibiotic resistance gene (ARG) variants, conferring resistance in pathogenic bacteria to tetracyclines, fluoroquinolones (-type) and macrolides (-type, -type and -type). Among these, -type was the most prevalent gene, found across 11 cyanobacterial genomes from the Nostocales order. presented the highest variety of close ARG matches, with hits for the macrolide resistance genes -type, -type and type. An analysis of the genomic assemblies also revealed an additional 12 ARGs in bacteria from the phyla Firmicutes, Proteobacteria and Bacteroidetes, present in the cyanobacterial cultures, foreseeing the horizontal gene transfer of ARGs with cyanobacteria. Additionally, more than 200 partial ARGs were detected on each recovered cyanobacterial genome, allowing for future studies of antibiotic resistance genotype/phenotype in cyanobacteria. These findings highlight the importance of further efforts to understand the role of cyanobacteria on the aquatic resistome from a One Health perspective.
- Recovery of cyanobacterial draft genomes from a metagenome dataset: applications to the detection of antibiotic resistance variantsPublication . Balata, DuarteFreshwater cyanobacteria are one of the most widely distributed groups of organisms in continental bodies of water worldwide. Cyanobacteria’s evolutionary process granted them a wide variety of shapes and forms of colony organization, making them a very heterogeneous group, both phenotypically and genotypically. Despite being a frequently studied group due to their capacity to form harmful algal blooms and produce bioactive compounds with promising pharmaceutical properties, their known potential as antibiotic resistance genes reservoirs is still understudied, especially in freshwater environments. Antibiotic resistance is a growing concern in terms of human and animal health, but antibiotic resistance mechanisms aren’t restricted to pathogenic bacteria. Environmental microorganisms have also been shown to play a key role in the emergence and propagation of antibiotic resistance genes. In this work, we use high-throughput sequencing technology (Illumina) to taxonomically classify cyanobacteria present in metagenomic samples, sequenced from non-axenic cultures of Portuguese surface freshwater reservoirs and wastewater treatment plants, belonging to Estela Sousa e Silva Algae Culture Collection and Blue Biotechnology and Ecotoxicology Culture Collection. Metagenomic reads from 43 sequenced samples of non-axenic cultures of cyanobacteria were filtered and assembled. Using a metagenome binning approach, we managed to isolate 41 cyanobacterial genomes and perform an analysis for the presence of antibiotic resistance genes, from the Comprehensive Antibiotic Resistance Database, on strains of Anabaena, Aphanizomenon, Microcystis aeruginosa, Planktothrix agardhii and Planktothrix mougeotii. We detected the presence of the fluoroquinolone and tetracycline resistance gene adeF across a multitude of samples from different genera and collection sites. The simultaneous occurrence of mutually potentiating macrolide resistance genes mphG and mefC, as previously described for marine bacteria, was also detected for the first time in cyanobacteria genomes. Copies of similar antibiotic resistance genes found across multiple bins from the same environmental samples suggests that horizontal gene transfer might be playing a role in the fast adaptation of bacteria to antimicrobial products. This work further corroborates the evidence for cyanobacteria’s importance in the freshwater resistome. In order to achieve these results, a GNU Make/ Shell script pipeline (CyanoPipeline) was created in order to increase the reproducibility of the work and facilitate the iterations of a complex workflow.