Yeasts from fermented Brazilian fruits as biotechnological tools for increasing phenolics bioaccessibility and improving the volatile profile in derived pulps.

Caatinga Biome fruits have been scarcely explored as a source of biotechnological yeasts. This study isolated yeasts from naturally fermented Caatinga fruits and evaluated Hanseniaspora opuntiae125,Issatchenkia terricola 129, and Hanseniaspora opuntiae 148 on fermentation of soursop and umbu-cajá pulps. All strains were able to ferment the pulps (72 h), increasing (p < 0.05) acetic acid, phenolics concentration and bioaccessibility, and maintaining counts above 7 log CFU/mL after fermentation and/or in vitro digestion. H. opuntiae 125 showed the highest counts (8.43-8.76 log CFU/mL; p < 0.05) in pulps and, higher organic acids production, increased survival to digestion, and higher bioaccessibility of various phenolics (p < 0.05) in the umbu-cajá pulp.I. terricola129 andH. opuntiae 148 showed higher metabolic activity, concentration and bioaccessibility of specific phenolics in umbu-cajá and soursop pulps, respectively (p < 0.05). Volatiles varied (p < 0.05) with the yeast strain. Generally, the yeast biotechnological performance for pulp fermentation was better on its fruit source.

Comparative toxicological evaluation of carvacrol, acetylcarvacrol anda fipronil-based pesticide in human blood cells.

Plant-derived chemicals are promising substances to control arthropod pests, although synthetic ones are still the most frequently used. Thus, comparative toxicological studies are needed to determine if natural substances are safe alternatives to replace the use of synthetic chemicals. This study aimed to compare the toxicity of carvacrol (natural origin), acetylcarvacrol (semi-synthetic) and a fipronil-based pesticide (synthetic). We assessed the effects of these chemicals on hemolytic activity, erythrocytes morphology and leucocyte viability using whole blood from human subjects. Additionally, DNA damage was evaluated through comet and DNA fragmentation assays. Fipronil and carvacrol caused hemolysis at concentrations ranging from 0.5 to 2.0%, whereas acetylcarvacrol did not cause hemolysis at 0.5 and 0.75%. Fipronil and carvacrol caused severe alterations in erythrocytes' morphology at 2%, such as ghost erythrocytes, elliptocyte-like shape and rouleau-like shape, presenting only 3.3 and 8.3% normal cells, respectively, at this concentration. However, 73.3% erythrocytes incubated with 2% acetylcarvacrol exhibited normal morphology. Fipronil considerably reduced leucocytes viability, decreasing it to 78% at 2%. Carvacrol and acetylcarvacrol showed no differences in leucocyte viability for 0.5 to 1.0%, but a decrease was observed for 2% carvacrol. The comet assay showed similar DNA damage for fipronil and carvacrol, but it was significantly lower for 1 and 2% acetylcarvacrol. Incubation with genomic DNA showed that only fipronil caused fragmentation of this molecule. Thus, we conclude that carvacrol and fipronil can present similar toxicity at higher concentrations. However, acetylation of carvacrol significantly reduced its toxicity to human blood cells compared with the other chemicals.

Phylogenomic characterization and pangenomic insights into the surfactin-producing bacteria Bacillus subtilis strain RI4914.

Bacillus subtilis is a versatile bacterial species able to produce surfactin, a lipopeptide biosurfactant. We carried out the phylogenomic characterization and pangenomic analyses using available B. subtilis complete genomes. Also, we report the whole genome of the biosurfactant-producing B. subtilis strain RI4914 that was isolated from effluent water from an oil exploration field. We applied a hybrid sequencing approach using both long- and short-read sequencing technologies to generate a highly accurate, single-chromosome genome. The pangenomics analysis of 153 complete genomes classified as B. subtilis retrieved from the NCBI shows an open pangenome composed of 28,511 accessory genes, which agrees with the high genetic plasticity of the species. Also, this analysis suggests that surfactin production is a common trait shared by members of this species since the srfA operon is highly conserved among the B. subtilis strains found in most of the assemblies available. Finally, increased surfactin production corroborates the higher srfAA gene expression in B. subtilis strain RI4914.

Evaluation of potentially probiotic yeasts and Lactiplantibacillus plantarum in co-culture for the elaboration of a functional plant-based fermented beverage.

This work aimed to evaluate the performance of co-cultivation of potential probiotic yeast and lactic acid bacteria (LAB) in producing plant-based fermented beverages. The co-culture comprised LAB Lactiplantibacillus plantarum CCMA0743 with the yeasts Pichia kluyveri CCMA 0615, Pichia guilliermondii CCMA 1753 and Debaryomyces hansenii CCMA 1761 separately. The plant substrate was 75 g oat, 175 g sunflower seeds, and 75 g almonds. The viability of microorganisms in the plant-based matrix was evaluated during fermentation, storage at 4 °C, and under simulated gastrointestinal tract (GIT) conditions. Chemical analysis, antioxidant activity, and sensory profile of the beverages were also determined. The three yeasts and the LAB showed counts greater than 6.0 log CFU/mL after fermentation, and the plant-based matrix protected the yeasts during simulated digestion. P. kluyveri and D. hansenii showed higher survival than P. guilliermondii and L. plantarum after exposure to simulated GIT conditions. The pH of the plant-based matrix reduced from approximately 7 to 3.8. Lactic acid was the main organic acid produced during fermentation. In addition, 113 volatile compounds were detected by gas chromatography-mass spectrometry (GC-MS), including alcohols, aldehydes, alkanes, alkenes, acids, ester, ether, ketones, phenol, and amides. The beverage sensory profile varied with the co-culture. The co-culture D. hansenii and L. plantarum showed higher antioxidant activity than the other co-culture tested, and the homogeneous texture attribute characterized the beverage produced with this combination. Results show the suitability of tested co-cultures to produce a plant-based fermented beverage and indicate more significant potential for D. hansenii and L. plantarum co-culture as a starter for its functionalization.

Probiotic Properties of Lactobacilli and Their Ability to Inhibit the Adhesion of Enteropathogenic Bacteria to Caco-2 and HT-29 Cells.

We evaluated the probiotic properties of lactic acid bacteria using resistance, safety, and functional assays. A preliminary subtractive screening of nineteen strains was performed based on their survival in simulated gastric and intestinal juice, and cell surface characteristics (hydrophobicity and auto-aggregation). Five strains were selected for further characterization, which included the assessment of their co-aggregation to pathogens, phenol tolerance, antimicrobial activity, and safety. Moreover, their adhesion to Caco-2 and HT-29 cells and the ability to inhibit pathogenic bacteria adhesion were evaluated. All strains had high (≥ 80.0%) survival rates in gastric and intestinal juices. Among them, Lactobacillus brevis CCMA 1284, L. plantarum CCMA 0743, and L. plantarum CCMA 0359 exhibited higher hydrophobicity (95.33, 96.06, and 80.02%, respectively), while L. paracasei CCMA 0504 and L. paracasei CCMA 0505 had the highest auto-aggregation values (45.36 and 52.66%, respectively). However, these last two strains were positive for the DNAse test, which is a safety concern. The CCMA 0359 and CCMA 1284 strains did not show antimicrobial activity, while the CCMA 0505 strain had a higher percentage of adhesion (4.75%) to Caco-2 cells. In the simulated competition and exclusion assays, the CCMA 0743 strain was able to reduce Salmonella adhesion to both cells (Caco-2 and HT-29), but only the CCMA 0743 and CCMA 0505 strains inhibited Escherichia coli adhesion to HT-29 cells in the competition assay. According to the results of these evaluated attributes, this strain showed to be an excellent candidate for probiotic use.