Eosinophils are granulocytes classically involved in allergic diseases and in the host immune responses to helminths, fungi, bacteria and viruses. The release of extracellular DNA traps by leukocytes is an important mechanism of the innate immune response to pathogens in various infectious conditions, including fungal infections. Aspergillus fumigatus is an opportunistic fungus responsible for allergic bronchopulmonary aspergillosis (ABPA), a pulmonary disease marked by prominent eosinophilic inflammation. Previously, we demonstrated that isolated human eosinophils release extracellular DNA traps (eosinophil extracellular traps; EETs) when stimulated by A. fumigatus in vitro. This release occurs through a lytic non-oxidative mechanism that involves CD11b and Syk tyrosine kinase. In this work, we unraveled different intracellular mechanisms that drive the release of extracellular DNA traps by A. fumigatus-stimulated eosinophils. Ultrastructurally, we originally observed that A. fumigatus-stimulated eosinophils present typical signs of extracellular DNA trap cell death (ETosis) with the nuclei losing both their shape (delobulation) and the euchromatin/heterochromatin distinction, followed by rupture of the nuclear envelope and EETs release. We also found that by targeting class I PI3K, and more specifically PI3Kδ, the release of extracellular DNA traps induced by A. fumigatus is inhibited. We also demonstrated that A. fumigatus-induced EETs release depends on the Src family, Akt, calcium and p38 MAPK signaling pathways in a process in which fungal viability is dispensable. Interestingly, we showed that A. fumigatus-induced EETs release occurs in a mechanism independent of PAD4 histone citrullination. These findings may contribute to a better understanding of the mechanisms that underlie EETs release in response to A. fumigatus, which may lead to better knowledge of ABPA pathophysiology and treatment.

Biodiversity is currently assessed for environmental characterizations and monitoring through a laborious and time-consuming process of morphological taxonomy. We used rRNA 18S, rRNA 28S and COI, together with NGS and Bioinformatics to identify benthic invertebrate organisms from sediment samples collected in five stations in the Campos Basin in southeast Brazil, an important oil extraction area and one of the best-studied marine biota in Brazil. A total of 3.3 million sequences were clustered in Operational Taxonomic Units and more than 1.6 million sequences (about 50% of all reads) were assigned to 957 prokaryotes and 577 eukaryotes. BLAST identified 23 phyla, 60 classes, 62 orders, 70 families, 67 genus and 46 species of eukaryotes. By meta-barcoding we identified phyla that are traditionally found in samples of marine benthos, such as Annelida, Arthropoda, Mollusca and Chordata, as well as rare phyla like Entoprocta and Gastrotricha. Taxa identified with meta-barcoding were compared to morphology data from previous studies in the area (REVIZEE, Habitats Project) and geo-validated with the database Global Biodiversity Information Facility. For several taxa, this is the first evidence of occurrence in Campos the area and the number of OTU identified suggests an enormous unveiled benthic biodiversity in Campos Basin. Our study supports the application of Meta-Barcoding for environmental characterization and monitoring programs, reducing from years to few months the time currently required for species identification and biodiversity determination.

The Atlantic Forest is one of the most import biodiversity hotspots in the world, nevertheless, its 20,000 plant species are poorly characterized genetically, what could undermine conservation efforts and bioprospection of natural products. We used a genome reduction using restriction site conservation (GR-RSC) technique to minimize sequencing effort and build in a short period a databank of gene sequences from 35 plant species from the Atlantic Forest in a private natural protected area in Southwest Brazil. After Illumina sequencing and standard bioinformatics, we produced more than 66 million super reads, of which 11 million (17%) were annotated using Diamond and UNIREF90 database and 55 million were 'No hit'. We picked 17 enzymes from 2 secondary metabolite synthesis pathways that are both important representatives of biological processes for plants and also of industrial interest, to test the usefulness of the databank we created for gene discovery. All 17 genes were detected in at least one of the 35 species and all species exhibited at least one of the genes. Eight of the 35 species exhibited all 17 genes. These results shows that genome simplification by restriction enzyme can be applied to preliminary screen thousands of species in tropical forests, generating useful databanks for scientific and entrepreneurial activities both in conservation biology and bioprospection.

Biodiversity is currently assessed for environmental characterizations and monitoring through a laborious and time-consuming process of morphological taxonomy. We used rRNA 18S, rRNA 28S and COI, together with NGS and Bioinformatics to identify benthic invertebrate organisms from sediment samples collected in five stations in the Campos Basin in southeast Brazil, an important oil extraction area and one of the best-studied marine biota in Brazil. A total of 3.3 million sequences were clustered in Operational Taxonomic Units and more than 1.6 million sequences (about 50% of all reads) were assigned to 957 prokaryotes and 577 eukaryotes. BLAST identified 23 phyla, 60 classes, 62 orders, 70 families, 67 genus and 46 species of eukaryotes. By meta-barcoding we identified phyla that are traditionally found in samples of marine benthos, such as Annelida, Arthropoda, Mollusca and Chordata, as well as rare phyla like Entoprocta and Gastrotricha. Taxa identified with meta-barcoding were compared to morphology data from previous studies in the area (REVIZEE, Habitats Project) and geo-validated with the database Global Biodiversity Information Facility. For several taxa, this is the first evidence of occurrence in Campos the area and the number of OTU identified suggests an enormous unveiled benthic biodiversity in Campos Basin. Our study supports the application of Meta-Barcoding for environmental characterization and monitoring programs, reducing from years to few months the time currently required for species identification and biodiversity determination.

Sediment fauna characterization and monitoring are mandatory requirements for obtaining oil and gas (O&G) environmental licensing for exploration and production (E&P) activities. Currently, for environmental characterizations and monitoring, biodiversity is assessed through morphological taxonomy, a time-consuming process. Taxonomists are constantly failing to meet the demands for biodiversity assessment required in monitoring programs. Thus, we combined three different phylogenetic markers(rDNA 18S, rDNA 28S and COI), HTS and Bioinformatics to identify benthic invertebrate organisms from sediment samples collected in five stations in the Campos Basin in southeast Brazil, an important oil extraction area and one of the best-studied marine biota in Brazil. Our results obtained with metagenomics were compared to morphology data provided by the Habitats Project whereas the database Global Biodiversity Information Facility ( www.gbif.org ) was used for organism localization. We obtained around 4.83 μg of DNA from 15 samples. A total of 3.3 million sequences were clustered in Operational Taxonomic Units and more than 1.6 million sequences (about 50% of all reads) were assigned to 957 prokaryotes and 577 eukaryotes. BLAST identified 23 phyla, 60 classes, 62 orders, 70 families, 67 genus and 46 species of eukaryotes. Our metagenomic analysis identified phyla that are traditionally found in samples of marine benthos, such as Annelida, Arthropoda, Mollusca and Chordata, as well as more rarely found phyla such as Bryozoa, Cnidaria, Echinodermata, Nematoda, Nemertea, Platyhelminthes, Porifera and Priapulida; and even more rare phyla like Entoprocta and Gastrotricha. The low availability of genetic markers for Brazilian species in Genebank impaired our ability to compare our findings with those obtained morphologically for which no sequences were found in Genebank. Our study shows that metagenomics can be applied for environmental characterization and monitoring programs and, with the possibility of automating the method, may reduce from years to few months the time currently required for species identification and biodiversity determination, which will certainly accelerate species discovery.