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Microorganisms Mar 2024(1) Background: The identification of microorganisms includes traditional biochemical methods, molecular biology methods evaluating the conserved regions of rRNA, and...
(1) Background: The identification of microorganisms includes traditional biochemical methods, molecular biology methods evaluating the conserved regions of rRNA, and the molecular biology of proteins (proteomics), such as MALDI-TOF MS mass spectrometry. This work aimed to identify the biodiversity of yeasts associated with stingless bee species' honey and pollen, , , and , from the region of São Gonçalo dos Campos-Bahia (BA) state, Brazil. (2) Methods: Cellular proteins were extracted from 2837 microbial isolates (pollen and honey) and identified via MALDI-TOF MS. The identified yeast species were also compared to the mass spectra of taxonomically well-characterized reference strains, available from the National Center of Biotechnology Information (NCBI) database. (3) Results: Nine yeast species were identified: , , , , , , , , and pollen had the highest number of yeast colonies. The yeasts and showed high populations in the samples of and , respectively. This work shows that there is some sharing of the same species of yeast between honey and pollen from the same beehive. (4) Conclusions: A total of 71.84% of the identified species present a high level of confidence at the species level. Eight yeast species (, , , , , , , and ) were found for the first time in the samples that the authors inspected. This contributes to the construction of new knowledge about the diversity of yeasts associated with stingless bee products, as well as to the possibility of the biotechnological application of some yeast species.
PubMed: 38674622
DOI: 10.3390/microorganisms12040678 -
Foods (Basel, Switzerland) Apr 2024An amplicon metagenomic approach based on the ITS2 region of fungal rDNA was used to investigate the diversity of fungi associated with mature strawberries collected...
An amplicon metagenomic approach based on the ITS2 region of fungal rDNA was used to investigate the diversity of fungi associated with mature strawberries collected from a volcanic orchard and open-air market stands. Based on the Kruskal-Wallis test, no statistically significant differences were observed in both non-phylogenetic and phylogenetic alpha diversity indices. According to beta diversity analyses, significant differences in fungal communities were found between groups (orchard vs. market). Taxonomic assignment of amplicon sequence variables (ASVs) revealed 7 phyla and 31 classes. The prevalent fungal phyla were (29.59-84.58%), (15.33-70.40%), and Fungi-phy-Insertae-sedis (0.45-2.89%). The most predominant classes among the groups were in the market group, and and in the orchard group. Based on the analysis of microbiome composition (ANCOM), we found that the most differentially fungal genera were , , and Endophytic yeasts were prevalent in both groups, while was detected in fruits originating only from the market. In addition, (relative abundance varying from 1.7% to 21.18%) and (relative abundance varying from 1.58% to 16.55%) were detected in all samples regardless of origin, while was detected in samples from the market only, their relative abundance varying with the sample (from 0.80% to 19.23%). Their role in fruit quality and safety has not been yet documented. Moreover, several clinically related yeasts, such as and , were detected in samples only from the market. Understanding the variety and makeup of the mycobiome in ripe fruits during the transition from the orchard to the market is crucial for fruit safety after harvest.
PubMed: 38672848
DOI: 10.3390/foods13081175 -
International Journal of Food... Jun 2024Previous investigations proved the potential of Saccharomyces cerevisiae MBELGA62 and Pichia kudriavzevii MBELGA61 as suitable biocontrolling agents against Aspergillus...
Previous investigations proved the potential of Saccharomyces cerevisiae MBELGA62 and Pichia kudriavzevii MBELGA61 as suitable biocontrolling agents against Aspergillus sp. through the production of soluble and volatile bioactive antifungal compounds. The present study delves into those finding by means of the identification of the volatile compounds produced by brewer's strains that demonstrated fungistatic and fungicidal effects against Aspergillus flavus and A. parasiticus when cultured in brewer's wort agar plates. Traditional brewer's yeasts such as S. cerevisiae MBELGA62 and Saccharomyces pastorianus SAFS235 synthetize volatiles that fully inhibited mycelial development for up to 9 days at 30 °C. The non-conventional brewer's strains P. kudriavzevii MBELGA61 and Meyerozyma guilliermondii MUS122 increased the lag phase by >100% and significantly reduced the fungal growth rate by 27.5-43.0% and 15.4-31.4%, respectively. In this context, 2-phenylethanol, 2-phenylethyl acetate and benzyl alcohol were identified as the main antifungal agents involved in Aspergillus sp.'s inhibition.
Topics: Aspergillus; Fermentation; Antifungal Agents; Volatile Organic Compounds; Saccharomyces cerevisiae; Pichia; Phenylethyl Alcohol
PubMed: 38640817
DOI: 10.1016/j.ijfoodmicro.2024.110692 -
World Journal of Microbiology &... Mar 2024Probiotic microorganisms are used to improve the health and wellness of people and the research on this topic is of current relevance and interest. Fifty-five yeasts,...
Probiotic microorganisms are used to improve the health and wellness of people and the research on this topic is of current relevance and interest. Fifty-five yeasts, coming from honeybee's ecosystem and belonging to Candida, Debaryomyces, Hanseniaspora, Lachancea, Metschnikowia, Meyerozyma, Starmerella and Zygosacchromyces genera and related different species, were evaluated for the probiotic traits. The resistance to gastrointestinal conditions, auto-aggregation, cell surface hydrophobicity or biofilm formation abilities as well as antimicrobial activity against common human pathogenic bacteria were evaluated. The safety analysis of strains was also carried out to exclude any possible negative effect on the consumer's health. The influence of proteinase treatment of living yeasts and their adhesion to Caco-2 cells were also evaluated. The greatest selection occurred in the first step of survival at the acidic pH and in the presence of bile salts, where more than 50% of the strains were unable to survive. Equally discriminating was the protease test which allowed the survival of only 27 strains belonging to the species Hanseniaspora guilliermondii, Hanseniaspora uvarum, Metschnikowia pulcherrima, Metschnikowia ziziphicola, Meyerozyma caribbica, Meyerozyma guilliermondii, Pichia kluyveri, Pichia kudriavzevii and Pichia terricola. An integrated analysis of the results obtained allowed the detection of seven yeast strains with probiotic aptitudes, all belonging to the Meyerozyma genus, of which three belonging to M. guillermondii and four belonging to M. caribbica species.
Topics: Bees; Animals; Humans; Ecosystem; Caco-2 Cells; Yeasts; Candida; Probiotics
PubMed: 38538981
DOI: 10.1007/s11274-024-03941-z -
Journal of Fungi (Basel, Switzerland) Mar 2024Yeast infections are challenging human and animal medicine due to low rates of detection and the emergence of unknown ecology isolates. The aim of this study was to...
Yeast infections are challenging human and animal medicine due to low rates of detection and the emergence of unknown ecology isolates. The aim of this study was to verify the biochemical identification of yeasts and yeast-like microorganisms obtained from animals comparing the results with chromogenic media and matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF MS). Between January and August 2023, yeast and yeast-like isolates from samples of animals with suspicion of mycosis were identified using Vitek 2 Compact, Brilliance Candida Agar and MALDI Biotyper MSP. A total of 39 cases were included, and 45 isolations were obtained. (15.5%, 7/45), (13.3%, 6/45), (11.1%, 5/45), and (8.9%, each one 4/45) were the most identified organisms. There was full agreement with the three identification methods in 71.1% (32/45) of the isolates, disagreement on species in 17.8% (8/45), disagreement on genus and species in 6.7% (3/45) and, in 4.4% (2/45), there was no matched pattern in MALDI-TOF to compare the results. Biochemical methods are a good option in laboratories where proteomics are not available, and chromogenic media enhances diagnostics by detecting mixed infections. Surveillance must be implemented to improve the detection of agents shared between humans and animals.
PubMed: 38535226
DOI: 10.3390/jof10030218 -
Synthetic and Systems Biotechnology Mar 2024Dopamine can be used to treat depression, myocardial infarction, and other diseases. However, few reports are available on the de novo microbial synthesis of dopamine...
Dopamine can be used to treat depression, myocardial infarction, and other diseases. However, few reports are available on the de novo microbial synthesis of dopamine from low-cost substrate. In this study, integrated omics technology was used to explore the dopamine metabolism of a novel marine multi-stress-tolerant aromatic yeast GXDK6. GXDK6 was found to have the ability to biosynthesize dopamine when using glucose as the substrate. 14 key genes for the biosynthesis of dopamine were identified by whole genome-wide analysis. Transcriptomic and proteomic data showed that the expression levels of gene encoding aspartate aminotransferase (regulating dopamine anabolism) were upregulated, while gene encoding copper amine oxidase (involved in dopamine catabolism) were downregulated under 10 % NaCl stress compared with non-NaCl stress, thereby contributing to biosynthesis of dopamine. Further, the amount of dopamine under 10 % NaCl stress was 2.51-fold higher than that of zero NaCl, which was consistent with the multi-omics results. Real-time fluorescence quantitative PCR (RT-qPCR) and high-performance liquid chromatography (HPLC) results confirmed the metabolic model of dopamine. Furthermore, by overexpressing , AST enzyme activity was increased by 24.89 %, the expression of genes related to dopamine metabolism was enhanced, and dopamine production was increased by 56.36 % in recombinant GXDK6AAT2. In conclusion, GXDK6 could utilize low-cost carbon source to synthesize dopamine, and NaCl stress promoted the biosynthesis of dopamine.
PubMed: 38292761
DOI: 10.1016/j.synbio.2024.01.002 -
Applied Microbiology and Biotechnology Jan 2024Honeybee (Apis mellifera) is an important agricultural pollinator and a model for sociality. In this study, a deep knowledge on yeast community characterizing the...
Honeybee (Apis mellifera) is an important agricultural pollinator and a model for sociality. In this study, a deep knowledge on yeast community characterizing the honeybees' environmental was carried out. For this, a total of 93 samples were collected: flowers as food sources, bee gut mycobiota, and bee products (bee pollen, bee bread, propolis), and processed using culture-dependent techniques and a molecular approach for identification. The occurrence of yeast populations was quantitatively similar among flowers, bee gut mycobiota, and bee products. Overall, 27 genera and 51 species were identified. Basidiomycetes genera were predominant in the flowers while the yeast genera detected in all environments were Aureobasidium, Filobasidium, Meyerozyma, and Metschnikowia. Fermenting species belonging to the genera Debaryomyces, Saccharomyces, Starmerella, Pichia, and Lachancea occurred mainly in the gut, while most of the identified species of bee products were not found in the gut mycobiota. Five yeast species, Meyerozyma guilliermondii, Debaryomyces hansenii, Hanseniaspora uvarum, Hanseniaspora guilliermondii, and Starmerella roseus, were present in both summer and winter, thus indicating them as stable components of bee mycobiota. These findings can help understand the yeast community as a component of the bee gut microbiota and its relationship with related environments, since mycobiota characterization was still less unexplored. In addition, the gut microbiota, affecting the nutrition, endocrine signaling, immune function, and pathogen resistance of honeybees, represents a useful tool for its health evaluation and could be a possible source of functional yeasts. KEY POINTS: • The stable yeast populations are represented by M. guilliermondii, D. hansenii, H. uvarum, H. guilliermondii, and S. roseus. • A. pullulans was the most abondance yeast detective in the flowers and honeybee guts. • Aureobasidium, Meyerozyma, Pichia, and Hanseniaspora are the main genera resident in gut tract.
Topics: Bees; Animals; Yeasts; Ascomycota; Pichia; Gastrointestinal Microbiome; Flowers
PubMed: 38276993
DOI: 10.1007/s00253-023-12942-1 -
Plants (Basel, Switzerland) Nov 2023Phytoremediation, an environmentally friendly and sustainable approach for addressing Cu-contaminated environments, remains underutilized in mine tailings. Arbuscular...
Phytoremediation, an environmentally friendly and sustainable approach for addressing Cu-contaminated environments, remains underutilized in mine tailings. Arbuscular mycorrhizal fungi (AMF) play a vital role in reducing Cu levels in plants through various mechanisms, including glomalin stabilization, immobilization within fungal structures, and enhancing plant tolerance to oxidative stress. Yeasts also contribute to plant growth and metal tolerance by producing phytohormones, solubilizing phosphates, generating exopolysaccharides, and facilitating AMF colonization. This study aimed to assess the impact of AMF and yeast inoculation on the growth and antioxidant response of plants growing in Cu mine tailings amended with compost. Plants were either non-inoculated (NY) or inoculated with (MG), (RM), or a combination of both (MIX). Plants were also inoculated with (CC), while others remained non-AMF inoculated (NM). The results indicated significantly higher shoot biomass in the MG-NM treatment, showing a 3.4-fold increase compared to the NY-NM treatment. The MG-CC treatment exhibited the most substantial increase in root biomass, reaching 5-fold that in the NY-NM treatment. Co-inoculation of AMF and yeast influenced antioxidant activity, particularly catalase and ascorbate peroxidase. Furthermore, AMF and yeast inoculation individually led to a 2-fold decrease in total phenols in the roots. Yeast inoculation notably reduced non-enzymatic antioxidant activity in the ABTS and CUPRAC assays. Both AMF and yeast inoculation promoted the production of photosynthetic pigments, further emphasizing their importance in phytoremediation programs for mine tailings.
PubMed: 38068648
DOI: 10.3390/plants12234012 -
Anais Da Academia Brasileira de Ciencias 2023The interest for oleaginous yeasts has grown significantly in the last three decades, mainly due to their potential use as a renewable source of microbial oil or single...
The interest for oleaginous yeasts has grown significantly in the last three decades, mainly due to their potential use as a renewable source of microbial oil or single cell oils (SCOs). However, the methodologies for cell disruption to obtain the microbial oil are considered critical and determinant for a large-scale production. Therefore, this work aimed to evaluate different methods for cell wall disruption for the lipid extraction of Yarrowia lipolytica QU21 and Meyerozyma guilliermondii BI281A. The two strains were separately cultivated in 5 L batch fermenters for 120 hours, at 26 ºC and 400 rpm. Three different lipid extraction processes using Turrax homogenizer, Ultrasonicator and Braun homogenizer combined with bead milling were applied in wet, oven-dried, and freeze-dried biomass of both strains. The treatment with the highest percentage of disrupted cells and highest oil yield was the ultrasonication of oven-dried biomass (37-40% lipid content for both strains). The fact that our results point to one best extraction strategy for two different yeast strains, belonging to different species, is a great news towards the development of a unified technique that could be applied at industrial plants.
Topics: Yarrowia; Oils; Biomass
PubMed: 38055604
DOI: 10.1590/0001-3765202320191256 -
Molecules (Basel, Switzerland) Oct 2023Using YB4, which was isolated and screened from southern Sichuan pickles in the laboratory, as the experimental group, we investigated the changes in growth, total...
Using YB4, which was isolated and screened from southern Sichuan pickles in the laboratory, as the experimental group, we investigated the changes in growth, total ester content, and volatile flavor substances of YB4 under different NaCl concentrations. The growth of YB4 was found to be inhibited by NaCl, and the degree of inhibition increased at higher NaCl concentrations. Additionally, the total ester content of the control group (CK) was significantly lower compared to the other groups ( < 0.05). The application of NaCl also resulted in distinct changes in the volatile profile of YB4, as evidenced by E-nose results. Gas chromatography-mass spectrometry (GC-MS) and gas chromatography-ion mobility spectrometry (GC-IMS) were employed to analyze the volatile compounds. A total of 148 and 86 volatiles were detected and identified using GC-MS and GC-IMS, respectively. Differential volatiles among the various NaCl concentrations in YB4 were determined by a variable importance in projection (VIP) analysis in partial least squares-discriminant analysis (PLS-DA). These differentially expressed volatiles were further confirmed by their relative odor activity value (ROAV) and odor description. Ten key contributing volatiles were identified, including ethanol, 1-pentanol, nonanal, octanal, isoamyl acetate, palmitic acid ethyl ester, acrolein, ethyl isobutanoate, prop-1-ene-3,3'-thiobis, and 2-acetylpyrazine. This study provides insights into the specificities and contributions of volatiles in YB4 under different NaCl concentrations. These findings offer valuable information for the development of aroma-producing yeast agents and the subsequent enhancement in the flavor of southern Sichuan pickles.
Topics: Gas Chromatography-Mass Spectrometry; Electronic Nose; Sodium Chloride; Volatile Organic Compounds; Odorants; Esters
PubMed: 37836821
DOI: 10.3390/molecules28196979