Luiz Gustavo dos Anjos Borges

Purpose Diabetes Mellitus (DM) can lead to significant reduction of saliva production, which increases the risk of oral diseases. Managing reduced salivary flow becomes indispensable for restoring microbial balance in this condition. This study aimed to investigate the effects of physical stimulation methods on salivary secretion rate and microbiome composition in individuals with type 2 DM (T2DM) having reduced salivary flow using an exploratory interim analysis of a randomized clinical trial. Methods Eight individuals provided a total of 15 stimulated or rest saliva samples and were randomly assigned to the following groups: low-level laser therapy (LLLT) (G1), transcutaneous electrical nerve stimulation (TENS)(G2), and a combination of both methods(G3), administered twice a week over a span of 10 sessions. Salivary flow was evaluated at three time points: recruitment, before the 1st session (baseline) and after the 10th session. Subjective dry mouth was evaluated using the TESS scale at baseline and after the final session. DNA was extracted from whole saliva immediately before treatment (baseline) and after the final stimulation session, 16 S rRNA gene amplified and sequenced (Illumina MiSeq). Results N = 7 individuals showed improvement in stimulated salivary flow (SSF), achieving or maintaining normal range (SSF > 0.7 ml/min). Bacterial diversity was significantly higher in saliva after stimuli with laser(G1) when compared to TENS(G2). The relative abundance of Firmicutes and Fusobacteriota increased after all treatments, while Actinobacteriota and Proteobacteria decreased. The abundance of minority taxa was reduced after stimulation. Conclusion This exploratory study provides initial indications that physical stimulation may help manage T2DM-related reduced salivary flow and could be associated with subtle changes in low-abundance taxa suggesting a potential “clearance” effect. These preliminary observations require confirmation through larger, long-term studies. Trial registration number RBR-3tqv8r3. Date of registration: 13th April 2023.

Mycolicibacterium gadium IBE100 and Mycobacterium paragordonae IBE200 are aerobic, chemoorganoheterotrophic bacteria isolated from activated sludge from a wastewater treatment plant. They use 2‐methylpropene (isobutene, 2‐MP) as the sole source of carbon and energy. Here, we postulate a degradation pathway of 2‐methylpropene derived from whole genome sequencing, differential expression analysis and peptide‐mass fingerprinting. Key genes identified are coding for a 4‐component soluble diiron monooxygenase with epoxidase activity, an epoxide hydrolase, and a 2‐hydroxyisobutyryl‐CoA mutase. In both strains, involved genes are arranged in clusters of 61.0 and 58.5 kbp, respectively, which also contain the genes coding for parts of the aerobic pathway of adenosylcobalamin synthesis. This vitamin is essential for the carbon rearrangement reaction catalysed by the mutase. These findings provide data for the identification of potential 2‐methylpropene degraders.

The aim of this study is to evaluate the effect of different thermal pretreatments of the inoculum on the diversity of the microbial community producing hydrogen from sugarcane vinasse. High-throughput sequencing of the 16S and 18S rRNA genes was performed. The reactor samples were also selected for the isolation of strict anaerobes. Decreased microbial diversity was observed with increasing pretreatment temperatures, with Firmicutes predominating: 90% to 97%. The highest abundance of Staphylococcus (7.9%) was found in pretreatment at 120 °C / 20 min at pH 6. The fungal analysis revealed a high prevalence of Candida (47%), Agaricomycetes, Pezizomycotina and Aspergillus in assays with higher H₂ production (90° C / 10 min at pH 6). Three species of Clostridium were isolated: C. bifermentans, C. saccharoperbutylacetonicum and C. saccharobutylicum. The isolates were tested separately and in co-cultures for the production of hydrogen. Hydrogen-producing capacity by co-culture of Clostridium species was increased by 18%. Knowing microorganisms and understanding the interaction between eukaryotes and prokaryotes is essential to obtain strategies for biotransformation of vinasse for the production of bioenergy.

Theactive sludge process is one of the most-used techniques for the biodegradation of organic compounds present in effluents from an assortment of wastewaters. This study investigated the bacterial community structure of a petroleum industry’s activated sludge and its physical and chemical parameters using high-throughput sequencing. Samples were collected over one year: autumn 2015 (C1), winter 2015 (C2), spring 2015 (C3), and summer 2016 (C4). Total DNA was extracted, and the primers targeting the V4 region of the 16S rRNA gene were used for amplicon sequencing. The majority of the detected microorganisms were considered rare microbiota, presenting a relative abundance below 1% of the total sequences. All of the sequences were classified at the phylum level, and up to 55% of the ASVs (Amplicon Sequence Variants) were associated with known bacterial genera. Proteobacteria was the most abundant phylum in three seasons, while the phylum Armatimonadota dominated in one season. The genus Hyphomicrobium was the most abundant in autumn, winter and summer, and an ASV belonging to the family Fimbriimonadaceae was the most abundant in the spring. Canonical Correspondence Analysis showed that physicochemical parameters of SS, SD and TSS are correlated, as well as ammoniacal nitrogen. Sample C3 presented the highest values of COD, AN and solids (SS, SD and TSS). The highest COD, AN, and solids values are correlated to the high frequency of the phylum Armatimonadota in C3.

Conventional agricultural practices, such as rice plantations, often contaminate the soil and water with xenobiotics. Here we evaluated the microbiota composition in experimental rice planting with a record of prolonged pesticide use, using 16S and 18S rRNA amplicon sequencing. We investigated four components of a complete agricultural system: affluent water (A), rice rhizosphere soil (R), sediment from a storage pond (S), and effluent (E) water (drained from the storage pond). Despite the short spatial distance between our sites, the beta diversity analysis of bacterial communities showed two well-defined clusters, separating the water and sediment/rhizosphere samples; rhizosphere and sediment were richer while the effluent was less diverse. Overall, the site with the highest evenness was the rhizosphere. Unlike the bacterial communities, Shannon diversity of microeukaryotes was significantly different between A and E. The effluent presented the lowest values for all ecological indexes tested and differed significantly from all sampled sites, except on evenness. When mapped the metabolic pathways, genes corresponding to the degradation of aromatic compounds, including genes related to pesticide degradation, were identified. The most abundant genes were related to the degradation of benzoate. Our results indicate that the effluent is a selective environment for fungi. Interestingly, the overall fungal diversity was higher in the affluent, the water that reached the system before pesticide application, and where the prokaryotic diversity was the lowest. The affluent and effluent seem to have the lowest environmental quality, given the presence of bacteria genera previously recorded in environments with high concentrations of pesticide residues. The microbiota, environmental characteristics, and pesticide residues should be further studied and try to elucidate the potential for pesticide degradation by natural consortia. Thus, extensive comparative studies are needed to clarify the microbial composition, diversity, and functioning of rice cultivation environments, and how pesticide use changes may reflect differences in microbial structure.

Live visualization of influenza A virus (IAV) structural proteins during viral infection in cells is highly sought objective to study different aspects of the viral replication cycle. To achieve this, we engineered an IAV to express a Tetra Cysteine tag (TC tag) from hemagglutinin (HA), which allows intracellular labeling of the engineered HA protein with biarsenic dyes and subsequent fluorescence detection. Using such constructs, we rescued a recombinant IAV with TC tag inserted in HA, in A/Puerto Rico/8/1934(H1N1) background (HA-TC). This recombinant HA-TC tag reporter IAV was replication-competent; however, as compared to wild type PR8 IAV, it was attenuated in multicycle replication. We confirmed expression of TC tag and biarsenical labeling of HA by immunofluorescence assay in cells infected with an HA-TC tag reporter IAV. Further, we used this reporter virus to visualize HA expression and translocation in IAV infected cells by live confocal imaging. We also tested the utility of the HA-TC IAV in testing chemical inhibitors of the HA translocation. Overall, HA-TC IAV is a versatile tool that will be useful for studying viral life cycle events, virus-host interactions, and anti-viral testing.

The chemical properties, ecotoxicity, and microbiome of leachates from phytomanaged Cu-contaminated soils were analyzed. The phytomanagement was carried out using Cu-tolerant poplar Populus trichocarpa × deltoides cv. Beaupré and black bent Agrostis gigantea L., aided by soil amendments, i.e., dolomitic limestone (DL) and compost (OM), alone and in combination (OMDL). Plants plus either DL or OMDL amendments reduced in leachates the electrical conductivity, the Cu concentration, and the concentration of total organic C except for the OMDL treatment, and decreased leachate toxicity towards bacteria. Total N concentration increased in the OM leachates. The aided phytostabilization increased the culturable bacteria numbers and the proportion of Cu-resistant bacteria in the leachates, as compared to the leachate from the untreated soil. Phytomanagement also enriched the microbial communities of the leachates with plant beneficial bacteria. Overall, the Cu stabilization and phytomanagement induced positive changes in the microbial communities of the soil leachates.

The ecology of complex microhabitats remains poorly characterized in most tropical and subtropical biomes, and holds potential to help understand the structure and dynamics of different biodiversity components in these ecosystems. We assessed nutritional and metabolic parameters of two bromeliad species (Aechmea gamosepala and Vriesea platynema) at an Atlantic Forest site and used 16S rDNA metabarcoding to survey the microbial communities inhabiting their tanks. We observed that levels of some nutrients (e.g., nitrogen) varied across seasons consistently in both species, while others (e.g., phenolic compounds) presented considerable differences between the two bromeliads. In contrast, patterns of tank microbial diversity did not follow a similar temporal trend. There was extensive variation in microbial composition among samples, which included intra-specific differences but also some consistent differences between the two bromeliads. For example, Citrobacter, Klebsiella and Pantoea presented significantly different abundances in the two species. Interestingly, the dominant bacterial genera in both species included Pseudomonas and Enterobacter, which have been reported to include plant-beneficial species. Overall, our data contribute to the characterization of the nutritional status of Atlantic Forest bromeliads and the composition of their prokaryotic communities, laying the foundation for detailed investigations targeting the ecological interactions between these plants and their associated microbes.