Deep Divergence and Genomic Diversification of Gut Symbionts of Neotropical Stingless Bees.

Social bees harbor conserved gut microbiotas that may have been acquired in a common ancestor of social bees and subsequently codiversified with their hosts. However, most of this knowledge is based on studies on the gut microbiotas of honey bees and bumblebees. Much less is known about the gut microbiotas of the third and most diverse group of social bees, the stingless bees. Specifically, the absence of genomic data from their microbiotas presents an important knowledge gap in understanding the evolution and functional diversity of the social bee microbiota. Here, we combined community profiling with culturing and genome sequencing of gut bacteria from six neotropical stingless bee species from Brazil. Phylogenomic analyses show that most stingless bee gut isolates form deep-branching sister clades of core members of the honey bee and bumblebee gut microbiota with conserved functional capabilities, confirming the common ancestry and ecology of their microbiota. However, our bacterial phylogenies were not congruent with those of the host, indicating that the evolution of the social bee gut microbiota was not driven by strict codiversification but included host switches and independent symbiont gain and losses. Finally, as reported for the honey bee and bumblebee microbiotas, we found substantial genomic divergence among strains of stingless bee gut bacteria, suggesting adaptation to different host species and glycan niches. Our study offers first insights into the genomic diversity of the stingless bee microbiota and highlights the need for broader samplings to understand the evolution of the social bee gut microbiota. IMPORTANCE Stingless bees are the most diverse group of the corbiculate bees and represent important pollinator species throughout the tropics and subtropics. They harbor specialized microbial communities in their gut that are related to those found in honey bees and bumblebees and that are likely important for bee health. Few bacteria have been cultured from the gut of stingless bees, which has prevented characterization of their genomic diversity and functional potential. Here, we established cultures of major members of the gut microbiotas of six stingless bee species and sequenced their genomes. We found that most stingless bee isolates belong to novel bacterial species distantly related to those found in honey bees and bumblebees and encoding similar functional capabilities. Our study offers a new perspective on the evolution of the social bee gut microbiota and presents a basis for characterizing the symbiotic relationships between gut bacteria and stingless bees.

Cultivable cellulolytic fungi isolated from the gut of Amazonian aquatic insects

Fungos filamentosos tem sido alvo de estudos de bioprospecção devido à sua grande eficiencia em produzir enzimas extracelulares, as quais tem grande potencial para uso em bioindústrias. Neste estudo, fungos filamentosos foram isolados do intestino de larvas de insetos aquáticos da Amazônia, para avaliar sua atividade celulolítica. Foram coletadas 69 larvas de insetos aquáticos fragmentadores de três gêneros: Phylloicus (Trichoptera: Calamoceratidae), Triplectides (Trichoptera:Leptoceridae) e Stenochironomus (Diptera: Chironomidae) em dez igarapés de uma área protegida na Amazônia central brasileira. O crescimento dos fungos isolados foi feito em meio de cultura Ágar Batata Dextrose (BDA). Os isolados fúngicos foram transferidos para o meio sintético com Carboximetil celulose e utilizou-se vermelho Congo para determinar o índice enzimático. O halo de hidrólise, indicando a produção de celulases, foi observado em 175 isolados fúngicos (70% do total), dos quais 25 tiveram um índice enzimático ≥ 2,0 e pertencem a sete gêneros fúngicos. Os táxons fúngicos Cladosporium, Gliocephalotrichum, Penicillium, Pestalotiopsis, Talaromyces, Trichoderma e Umbelopsis foram isolados dos intestinos das larvas de Phylloicus, Triplectides e Stenochironomus e são tradicionalmente utilizados em aplicações biotecnológicas. Os resultados indicam um potencial celulolítco destes fungos associados ao intestino de insetos aquáticos amazônicos. (AU)

The antimicrobial peptide HS-1 inhibits dengue virus infection.

Dengue virus (DENV) is an arbovirus that belongs to the Flaviviridae family. Studies reveal that peptides secreted by amphibians have many functions, such as antiviral and antimicrobial activities. As there is no antiviral drug effective against the DENV, the antiviral activity of a synthetic peptide called HS-1, derived from the secretion of the anuran Hypsiboas semilineatus, has been evaluated. The assays of neutralization in the Vero cells show a complete inhibition of infection of the serotypes 2 and 3. Furthermore, the direct action of peptides on the viral particle can be observed through atomic force microscopy. In vivo tests display 80% protection against the dengue-2 virus due to the presence of HS-1, which reveals its potential as an antiviral against the DENV.

Identification and characterization of an antimicrobial peptide of Hypsiboas semilineatus (Spix, 1824) (Amphibia, Hylidae).

The multidrug-resistant bacteria have become a serious problem to public health. In this scenery the antimicrobial peptides (AMPs) derived from animals and plants emerge as a novel therapeutic modality, substituting or in addition to the conventional antimicrobial. The anurans are one of the richest natural sources of AMPs. In this work several cycles of cDNA cloning of the skin of the Brazilian treefrog Hypsiboas semilineatus led to isolation of a precursor sequence that encodes a new AMP. The sequence comprises a 27 residue signal peptide, followed by an acidic intervening sequence that ends in the mature peptide at the carboxy terminal. The AMP, named Hs-1, has 20 amino acids residues, mostly arranged in an alpha helix and with a molecular weight of 2144.6 Da. The chemically synthesized Hs-1 showed an antimicrobial activity against all Gram-positive bacteria tested, with a range of 11-46 µM, but it did not show any effect against Gram-negative bacteria, which suggest that Hs-1 may have a selective action for Gram-positive bacteria. The effects of Hs-1 on bacterial cells were also demonstrated by transmission electron microscopy. Hs-1 is the first AMP to be described from H. semilineatus.