Biotech (Brazil)

Staphylococcus aureus is a Gram-positive bacterium living abundantly on our skin and mucous membranes. When there is an imbalance in microbiota, they are the main protagonists of various infections, such as soft tissue infections and bacteremia. However, Staphylococcus epidermidis also colonizes this microbiome, is able to compete with pathogenic bacteria, including methicillin-resistant Staphylococcus aureus (MRSA), and can contribute to treatments such as photodynamic inactivation (PDI) by inhibiting infection progression and restoring a healthy microbiota. In vitro photodynamic inactivation experiments were carried out using synthetic curcumin at a concentration of 5 μM as a photosensitizer and varying light doses (1, 2 and 5 J/cm2) at a wavelength of 450 nm, on pure cultures (S. aureus, S. epidermidis and MRSA) and mixed cultures, in which bacteria were placed together proportionally. This study revealed that pure cultures of these bacteria obtained statistically significant results with varying light doses of 2 and 5 J/cm2. In addition, in an attempt to bring infections closer to reality, experiments were carried out on mixed cultures. The results were not only significant but also increased reduction of bacteria, including resistant bacteria. Study offers new perspectives on the importance of themicrobiota for treatment of infections caused by the Staphylococcus genus.

Abstract Cancers are malignant neoplastic diseases with complex etiopathogenesis. Transformed cells acquire diverse biological properties that induce alterations in migration, invasion, and cell survival rates, favoring metastasis formation. This process is influenced by many factors, including miRNAs. MiRNAs are molecules that canonically prevent mRNA translation, influencing many cellular processes, carcinogenesis, and the progression of cancers. Nasopharyngeal carcinomas (NPCs) are aggressive cancers with high invasion rates to adjacent tissues, such as the base of the cranium, facial sinuses, and orbits, in addition to a high metastatic rate to local lymph nodes. The nonkeratinizing NPC is the most common histological form (over 95% of cases) and is highly related to Epstein-Barr virus (EBV) infection, notably when undifferentiated. In a previous study, we observed that immortalized nasopharyngeal cells NP69SV40Ttransfected to express the EBV Latent Membrane Protein 1 (LMP1) from the M81 strain hyperregulate human microRNAs miR-100-5p, miR-192-5p, and miR-574-3p, compared to NP69SV40Tcells transfected with LMP1 from the EBV strain B95.8A. In this study, we aimed to assess the putative effects of each of the identified microRNAs on the viability, migration, and proliferation of NP69SV40Tcells cultivated in vitro. Therefore, target prediction and pathway enrichment analysis were performed to infer which pathways could be modulated by each miRNA, followed by expression analysis of selected predicted target genes. Additionally, NP69SV40Tcells were transfected with miR-100-5p, miR-192-5p, or miR-574-3p mimics to analyze cell behavior. After their transfection with the miR-192-5p mimic, we observed a 40% decrease inRAB2Atranscript levels and a 43% reduction in cell migration. During the pathway enrichment analysis, miR-192-5p showed predicted involvement in pathways such as MAPK, GPCRs, TKR, and TGF-β. NP69SV40Tcells transfected with miR-100-5p or miR-574-3p reduced transcriptional levels of their targets FZD8 and STC1, respectively. However, in the performed cellular assays, they did not alter cell behavior. The results demonstrate that miR-192-5p acts as a tumor suppressor, elucidating part of its function in nasopharyngeal cells. This highlights its importance in the NPC context and the necessity of further studies on this miRNA.

This study proposes using the CRISPR transcriptional activation strategy to modulate the expression of genes encoding defense proteins and antimicrobial peptides (AMPs) in Phaseolus vulgaris. Three genes (PvD1, Pv-thionin, and Pv-lectin) were selected and targeted by the CRISPR-dCas9-TV-mediated transcriptional activation complex in the P. vulgaris L. hairy root. RT-qPCR investigated their activation efficiency. The eGFP-positive transgenic hairy roots exhibit enhanced expression of targeted genes compared to that of control roots. A moderate increase of 1.37-fold in PvD1 gene expression was observed in transgenic hairy roots, while 6.97-fold (Pv-lectin) and 5.70-fold (Pv-thionin) increases were observed. Importantly, no off-target effects of sgRNAs were detected, ensuring the precision and safety of the CRISPR-dCas9-TV strategy. The present article is a proof-of-concept study, and it has succeeded in demonstrating the efficiency of the CRISPR-dCas9-TV strategy in modulating the expression of target genes in P. vulgaris, paving the way for an alternative approach to protecting such essential crop plants.

The Thumbless bat Furipterus horrens (Cuvier, 1828) (Chiroptera: Furipteridae) is a tiny insectivorous bat species that has not undergone comprehensive systematic revisions, as taxonomic studies about this species are only represented by the middle XIX century description of F. caerulescens Tomes, 1856. Concurrently there also has been no assessment on the genetic diversity within the species along its distribution. We analyzed patterns of mitochondrial genetic variation of F. horrens under the prediction that structured populations could be associated with the naturally patchy distribution of these bats’ roosts. Our data revealed highly divergent lineages, with low haplotype sharing and indicating well-structured populations. For the phylogeny we recovered well-supported clades, and the outcome of species limits tests suggests the split of F. horrens into a complex of four to six putative species. Lineages recovered were mostly allopatric along South America, except for two sympatric populations from the low-medium Xingu River. Based on the outstandingly high genetic discontinuities found for Furipterus, we hypothesize that historical metapopulational arrangements composed of spatially structured populations with different degrees of isolation resulted in limited gene flow for at least part of the lineages found. The observed results may reflect a female-inherited variation at least partially related to yet unknown biological traits of Furipterus such as social structure, female philopatry or other biological characteristics. Whether part of these lineages represent separate full species needs further examination based on independent datasets (morphology and nuclear data) but it is now clear that Furipterus horrens is composed of distinct populations likely containing more than one species, a diversity that has remained virtually overlooked for over two centuries.

ABSTRACTWhite mould is a disease caused by the fungus Sclerotinia sclerotiorum, and it is considered one of the most devastating diseases in soybean crops, causing huge losses in productivity. In this study, we aimed to contribute to the genetic control of this disease by characterising the phenotypic reaction of 67 Brazilian soybean genotypes to S. sclerotiorum under controlled conditions. Plants were artificially inoculated with mycelium discs when they reached the four‐node phenological stage. Symptoms of white mould developed in all soybean genotypes. The resistance reaction was characterised by measuring the length of lesions on the main stem at 7 days after inoculation. Of the 67 genotypes evaluated, nine showed greater resistance to S. sclerotiorum, including 17S‐01443‐L8, 16S‐00630‐L4, GER_00003, 13S‐00001‐L2/FPS 2457 RR, BMX Torque I2X, P95Y02 IPRO, 17S‐00842‐L5, HO Pirapó IPRO and TMG 2359 IPRO. Twenty‐three genotypes showed an intermediate response, while 35 genotypes showed greater susceptibility to S. sclerotiorum. Further studies should be conducted under field conditions with the soybean genotypes that showed greater resistance response to S. sclerotiorum under controlled conditions.

Raw reads and consensus fasta sequences from coinfection cases are available in the GISAID database under accession codes: EPI_ISL_18907330, EPI_ISL_18907331, EPI_ISL_18907332, EPI_ISL_18907333, EPI_ISL_18907334, and EPI_ISL_18907335. Datasets used in phylogenetic analysis, major and minor consensus sequences, and genome assembly metrics are available on GitHub (https://github.com/salvatolab/coinfection-Dengue-clades-Southern-Brazil).

Research into microbial interactions during coffee processing is essential for developing new methods that adapt to climate change and improve flavor, thus enhancing the resilience and quality of global coffee production. This study aimed to investigate how microbial communities interact and contribute to flavor development in coffee processing within humid subtropical climates. Employing Illumina sequencing for microbial dynamics analysis, and high-performance liquid chromatography (HPLC) integrated with gas chromatography-mass spectrometry (GC-MS) for metabolite assessment, the study revealed intricate microbial diversity and associated metabolic activities. Throughout the fermentation process, dominant microbial species included Enterobacter, Erwinia, Kluyvera, and Pantoea from the prokaryotic group, and Fusarium, Cladosporium, Kurtzmaniella, Leptosphaerulina, Neonectria, and Penicillium from the eukaryotic group. The key metabolites identified were ethanol, and lactic, acetic, and citric acids. Notably, the bacterial community plays a crucial role in flavor development by utilizing metabolic versatility to produce esters and alcohols, while plant-derived metabolites such as caffeine and linalool remain stable throughout the fermentation process. The undirected network analysis revealed 321 interactions among microbial species and key substances during the fermentation process, with Enterobacter, Kluyvera, and Serratia showing strong connections with sugar and various volatile compounds, such as hexanal, benzaldehyde, 3-methylbenzaldehyde, 2-butenal, and 4-heptenal. These interactions, including inhibitory effects by Fusarium and Cladosporium, suggest microbial adaptability to subtropical conditions, potentially influencing fermentation and coffee quality. The sensory analysis showed that the final beverage obtained a score of 80.83 ± 0.39, being classified as a specialty coffee by the Specialty Coffee Association (SCA) metrics. Nonetheless, further enhancements in acidity, body, and aftertaste could lead to a more balanced flavor profile. The findings of this research hold substantial implications for the coffee industry in humid subtropical regions, offering potential strategies to enhance flavor quality and consistency through controlled fermentation practices. Furthermore, this study contributes to the broader understanding of how microbial ecology interplays with environmental factors to influence food and beverage fermentation, a topic of growing interest in the context of climate change and sustainable agriculture.

Second generation biorefineries play an important role in the production of renewable energy and fuels, utilizing forest and agro-industrial residues and by-products as raw materials. The integration of novel bioproducts, such as: xylitol, β-carotene, xylooligosaccharides, and biopigments into the biorefinery’s portfolio can offer economic benefits in the valorization of lignocellulosic materials, particularly cellulosic and hemicellulosic fractions. Fungal biopigments, known for their additional antioxidant and antimicrobial properties, are appealing to consumers and can have applications in various industrial sectors, including food and pharmaceuticals. The use of lignocellulosic materials as carbon and nutrient sources for the growth medium helps to reduce production costs, increasing the competitiveness of fungal biopigments in the market. In addition, the implementation of biopigment production in biorefineries allows the utilization of underutilized fractions, such as hemicellulose, for value-added bioproducts. This study deals with the potential of fungal biopigments production in second generation biorefineries in order to diversify the produced biomolecules together with energy generation. A comprehensive and critical review of the recent literature on this topic has been conducted, covering the major possible raw materials, general aspects of second generation biorefineries, the fungal biopigments and their potential for incorporation into biorefineries.