Search Results (53)

The comprehensive analysis of microbial communities reveals the unique microbial identity of different olive varieties, paving the way for new strategies in their development and commercial exploitation. In this context, the present study aimed to explore the microbial diversity and functional characteristics of Tsounati variety olives from the Monemvasia region of Peloponnese, Greece, that were naturally fermented for three months. The bacterial and fungal microbiota of both olives and brines were fingerprinted throughout the fermentation through classical microbiological analysis combined with molecular techniques. Among the 148 isolated bacteria, 85 were lactic acid bacteria (LAB), and 63 belonged to the Enterobacteriaceae family, while the 178 fungal isolates comprised 136 yeasts and 42 non-yeast or yeast-like fungi. Metataxonomic analysis confirmed the dominance of the bacterial genera Lactiplantibacillus, Leuconostoc, along with the Enterobacteriaceae family, and it revealed the presence of Coleofasciculaceae cyanobacteria mostly in olives. The dominant fungal genera were yeasts, namely Saccharomyces, Nakazawaea, and Cyberlindnera. Using the Folin–Ciocalteu assay, the average total polyphenol content of Tsounati fermented olive samples was 761.80 ± 128.87 mg gallic acid equivalents kg⁻¹ after 90 days of fermentation. The concentrations of the triterpenic, maslinic, and oleanolic acids, as determined by HPLC, remained stable throughout fermentation, with average values of 4764 and 1807 mg kg⁻¹, respectively. Finally, sensory analysis revealed the rich aromatic character of Tsounati variety, highlighting its potential to be used for Greek-style table olive production. Full article

The Demarcated Region of Madeira (DRM) is one of the oldest wine regions in Portugal, where the famous Madeira Wine (MW) is produced by spontaneous fermentation using endogenous yeasts. Several studies reported the role of endogenous Saccharomyces cerevisiae strains in the regional identity of wines, but only a few studies have been published in the DRM. The PCR-Interdelta (Polymerase Chain Reaction-Interdelta) analysis is a reliable method for S. cerevisiae strain identification. Here, we report the S. cerevisiae strains isolated from six Vitis vinifera grape varieties, namely, Tinta Negra, Boal, Sercial, Verdelho, Malvasia de São Jorge, and Complexa, which are widely used in MW production. During the 2020 campaign, eleven samples were collected from licensed vineyards and a winery, and submitted to spontaneous microfermentations and yeast isolation. Of the 1452 isolates counted, 1367 (94.2%) presented morphological characteristics of S. cerevisiae. We randomly selected 330 isolates from the positive colonies for strain identification. First, the PCR-Interdelta was optimized in ten commercial strains, using δ2–δ12 and δ12–δ21 pairs of primers, and δ2–δ12 primers were selected to screen the 330 isolates. We detected three fermentative profiles and a total of 25 PCR-Interdelta patterns were obtained, representing 7.6% of intraspecific variability, starting with the first non-official collection. The findings underscore the pivotal role of S. cerevisiae strain diversity in shaping the regional identity and quality of wines, with molecular tools like PCR-Interdelta analysis proving essential for monitoring intraspecific variability. Full article

Previous studies have shown the positive effects of non-conventional Metschnikowia spp. yeasts in mixed cultures with Saccharomyces cerevisiae on the properties of fruit wines. In this study, we investigated the effects of using conventional S. cerevisiae and non-conventional Metschnikowia pulcherrima yeasts as starter cultures in controlled mixed fermentations of honey wort. Other non-conventional yeasts were also tested for comparison, including Wickerhamomyces anomalus, Dekkera/Bretannomyces bruxellensis, and Wickerhamomyces anomalus. We evaluated the tolerance of the tested yeasts to high sugar content and analyzed the metabolic profiles of both monocultures and mixed systems. The M. pulcherrima strain showed the highest tolerance to 30% w/v glucose. The chemical complexity of fermented honey was improved using M. pulcherrima in co-starters with S. cerevisiae. The fermented honey samples were characterized by lower ethanol content, higher glycerol level, and rich volatilomes containing higher levels of both esters (ethyl acetate, 3-methylbutyl acetate, 2-methylpropyl acetate) and aliphatic alcohols (2-methylpropan-1-ol, 3-methylbutan-1-ol, and 2-methylbutan-1-ol). Similar characteristics were obtained using mixed populations of four strains: S. cerevisiae, M. pulcherrima, D. bruxellensis, and W. anomalus. Full article

A new generation of food packaging materials, centered on green solutions, is currently being developed in labs basing these materials on underused secondary industrial food by-products which have the ability to reduce the amount of petroleum-based packaging generated in order to minimize environmental harm and food by-products while ensuring food quality and safety. This study presents a sustainable biopolymeric combination based on bacterial nanocellulose grafted in yeast films, its potential to improve matrices properties, and the influence of plasticizer and emulsifier concentrations on mechanical properties, volatile fingerprint, and antimicrobial activity of films. Yeast films with 1.00% glycerol and 1.00% Tween 80, functionalized with a 2.00% mixture of cinnamon–lavender essential oils and with 2.00% bacterial cellulose produced from SCOBY presented improved mechanical properties compared to the control and exerted antipathogenic activity against Listeria monocytogens, Staphylococcus aureus, and Escherichia coli. Yeast films with grafted bacterial cellulose could be a sustainable food packaging solution for future applications. Full article

Background: The role of the S. cerevisiae strain in defining the volatile fingerprint is expressed throughout alcoholic fermentation and post-fermentation sur lie aging and is crucial for customizing the wine style. Methods: In this study, the alcoholic fermentation was carried out in a synthetic must to exclusively bring out the performance of the yeast in terms of volatile compound production, excluding the effect of the grape. Results: Among the 33 volatile organic compounds identified in the synthetic wines by GC-MS, esters, alcohols, and acids, represented the major groups for the nine different commercial oenological strains tested. All the relevant differences in the volatile fingerprint of the synthetic wines, which were lab-scale fermented, were quantitative rather than qualitative. The clustergram representation of the volatiles revealed an outstanding fingerprint for two strains (VIN13 and VIN7) among those tested, featuring hexanoic acid, octanoic acid, the corresponding esters (ethyl hexanoate, ethyl octanoate), and the acetates (2-phenylethyl acetate, isoamyl acetate), all at the highest levels. No relationship was appreciated between the fermentation rate and the volatile fingerprints. Conclusions: The outcomes of this study address the wine industry’s needs, supplying a full characterization of a broad range of commercial yeasts’ ability in fermentative volatile production. Full article

In 2019, Candida auris became the first fungal pathogen included in the list of the urgent antimicrobial threats by the Centers for Disease Control (CDC). Short tandem repeat (STR) analysis and whole-genome sequencing (WGS) are considered the gold standard, and can be complemented by other molecular methods, for the genomic surveillance and clade classification of this multidrug-resistant yeast. However, these methods can be expensive and require time and expertise that are not always available. The long turnaround time is especially not compatible with the speed needed to manage clonal transmission in healthcare settings. Fourier transform infrared (FTIR) spectroscopy, a biochemical fingerprint approach, has been applied in this study to a set of 74 C. auris isolates belonging to the five clades of C. auris (I-V) in combination with an artificial neural network (ANN) algorithm to create and validate “CLADE-FINDER”, a tool for C. auris clade determination. The CLADE-FINDER classifier allowed us to discriminate the four primary C. auris clades (I-IV) with a correct classification for 96% of the samples in the validation set. This newly developed genotyping scheme can be reasonably applied for the effective epidemiological monitoring and management of C. auris cases in real time. Full article

As a result of climate change, the phenology of grapes has been altered, mainly by increasing the sugar content and decreasing the acidity of ripe grapes. This shift, when the must is fermented, affects the quality of the wine. In this regard, the use of selected Saccharomyces and non-Saccharomyces yeasts to mitigate these undesirable effects in wine fermentations entails new strategies to improve their control and also to obtain wines better adapted to current consumer preferences. This work focuses on the use of a commercially available strain of Lachancea thermotolerans immobilized in biological support to form “microbial biocapsules”, comparing its effect with a free format and spontaneous fermentation on alcoholic fermentation and volatile compound composition. These biocapsules, consisting of yeast cells attached to fungal pellets, are being tested to improve wine sensory attributes and also to facilitate yeast inoculation in fermentative and clarification winemaking processes, as well as to reduce time and production costs. The composition of young wines obtained with L. thermotolerans, inoculated as free or biocapsule formats, were compared with those obtained by the traditional method of spontaneous fermentation using native yeast by quantifying 12 oenological variables and the contents in 12 major volatiles, 3 polyols, and 46 minor volatile compounds. The analytical data matrix underwent statistical analysis to compare and establish significant differences at p ≤ 0.05 level between the different wines obtained. Among the major volatiles and polyols, only ethyl acetate, 1,1-diethoxyethane, methanol, 2-methyl-1-butanol, acetoin, ethyl lactate, and glycerol showed significant differences in L. thermotolerans wines. Also, from the minor volatile metabolites, eight showed a significant dependence on the format used for L. thermotolerans, and the other nine volatiles were dependent on both yeast and inoculation format. Only 27 volatiles were selected as aroma-active compounds with odor activity values higher than 0.2 units. Statistical analysis showed a clear separation of the obtained wines into groups when subjected to Principal Component Analysis, and the fingerprinting of wines made with biocapsules shows intermediate values between the two remaining inoculation formats, particularly in the fruity/ripe fruit, green, and floral series. The organoleptic evaluation of wines results in significantly higher values in taste, overall quality, and total score for wines obtained with biocapsules. Full article

Kombucha is a fermented beverage traditionally made from the leaves of Camelia sinensis. The market has drastically expanded recently, and the beverage has become more elaborated with new, healthy food materials and flavors. Pruning and harvesting during coffee production may generate tons of coffee leaves that are discarded although they contain substantial amounts of bioactive compounds, including those found in maté tea and coffee seeds. This study characterized the changes in volatilome, microbial, and sensory profiles of pure and blended arabica coffee leaf tea kombuchas between 3–9 days of fermentation. Acceptance was also evaluated by consumers from Rio de Janeiro (n = 103). Kombuchas (K) were prepared using black tea kombucha starter (BTKS) (10%), sucrose (10%), a symbiotic culture of Bacteria and Yeasts (SCOBY) (2.5%), and a pure coffee leaf infusion (CL) or a 50:50 blend with toasted maté infusion (CL-TM) at 2.5%. The RATA test was chosen for sensory profile characterization. One hundred volatile organic compounds were identified when all infusions and kombucha samples were considered. The potential impact compounds identified in CL K and CL-TM K were: methyl salicylate, benzaldehyde, hexanal, nonanal, pentadecanal, phenylethyl-alcohol, cedrol, 3,5-octadien-2-one, β-damascenone, α-ionone, β-ionone, acetic acid, caproic acid, octanoic acid, nonanoic acid, decanoic acid, isovaleric acid, linalool, (S)-dihydroactinidiolide, isoamyl alcohol, ethyl hexanoate, and geranyl acetone. Aroma and flavor descriptors with higher intensities in CL K included fruity, peach, sweet, and herbal, while CL-TM K included additional toasted mate notes. The highest mean acceptance score was given to CL-TM K and CL K on day 3 (6.6 and 6.4, respectively, on a nine-point scale). Arabica coffee leaf can be a co-product with similar fingerprinting to maté and black tea, which can be explored for the elaboration of potentially healthy fermented beverages in food industries. Full article

Low fermentation temperatures are usually employed to obtain high-quality wines. This is especially interesting for white wine production since it prevents the loss of volatile compounds and a browning appearance; however, available fermentative yeasts do not usually tolerate low temperatures. Therefore, an interesting place to find new yeasts with cryotolerance is the Antarctic continent. From soil samples collected in Antarctica, 125 yeasts were isolated, of which 25 exhibited fermentative activity at 10 °C. After a fingerprinting assay, we classified the candidates into nine isotypes and sequenced internal transcribed spacer regions for their identification. These yeasts were identified as part of the Mrakia genus. Sugar and alcohol tolerance tests showed that some of these Antarctic soil yeasts were able to grow up to 9% alcohol, and 25% sugar was reached; however, they exhibited longer latency periods compared to the control Saccharomyces cerevisiae. The optimal growing temperature for the isolated Antarctic yeasts was between 10 °C and 15 °C. A comprehensive analysis of the results obtained showed that the isolates 10M3-1, 4M3-6, and 4B1-35 could be good candidates for fermentation purposes due to their alcohol, sugar tolerance, and growth features. Our results prove that it is possible to isolate fermentative yeasts from Antarctic soil with promising characteristics for their potential use in the wine production industry. Full article

In recent years, there has been renewed interest in the maintenance of food quality and food safety on the basis of metabolomic fingerprinting using vibrational spectroscopy combined with multivariate chemometrics. Nontargeted spectroscopy techniques such as FTIR, NIR and Raman can provide fingerprint information for metabolomic constituents in agricultural products, natural products and foods in a high-throughput, cost-effective and rapid way. In the current review, we tried to explain the capabilities of FTIR, NIR and Raman spectroscopy techniques combined with multivariate analysis for metabolic fingerprinting and profiling. Previous contributions highlighted the considerable potential of these analytical techniques for the detection and quantification of key constituents, such as aromatic amino acids, peptides, aromatic acids, carotenoids, alcohols, terpenoids and flavonoids in the food matrices. Additionally, promising results were obtained for the identification and characterization of different microorganism species such as fungus, bacterial strains and yeasts using these techniques combined with supervised and unsupervised pattern recognition techniques. In conclusion, this review summarized the cutting-edge applications of FTIR, NIR and Raman spectroscopy techniques equipped with multivariate statistics for food analysis and foodomics in the context of metabolomic fingerprinting and profiling. Full article

Beauveria bassiana is a dimorphic and entomopathogenic fungus with different ecological roles in nature. In pathogenic fungi, yeast-to-mycelial conversion, which is controlled by environmental factors, is required for virulence. Here, we studied the effects of different stimuli on the morphology of two B. bassiana strains and compared the toxicities of culture filtrates. In addition, we explored the role of volatiles as quorum sensing-like signals during dimorphic transition. The killing assays in Caenorhabditis elegans (Nematoda: Rhabditidae) showed that strain AI2 isolated from a mycosed insect cadaver had higher toxicity than strain AS5 isolated from soil. Furthermore, AI2 showed earlier yeast-to-mycelial switching than AS5. However, an increase in inoculum size induced faster yeast-to-mycelium conversion in AS5 cells, suggesting a cell-density-dependent phenomenon. Gas chromatography-mass spectrometry (GC-MS) analyses showed that the fingerprint of the volatiles was strain-specific; however, during the morphological switching, an inverse relationship between the abundance of total terpenes and 3-methylbutanol was observed in both strains. Fungal exposure to 3-methylbutanol retarded the yeast-to-mycelium transition. Hence, this study provides evidence that volatile compounds are associated with critical events in the life cycle of B. bassiana. Full article

The increasing number of infections caused by antimicrobial multi-resistant microorganisms has led to the search for new microorganisms capable of producing novel antibiotics. This work proposes Streptomyces pakalii sp. nov. as a new member of the Streptomycetaceae family. The strain ENCB-J15 was isolated from the jungle soil in Palenque National Park, Chiapas, Mexico. The strain formed pale brown, dry, tough, and buried colonies in the agar with no diffusible pigment in GAE (glucose–asparagine–yeast extract) medium. Scanning electron micrographs showed typical mycelium with long chains of smooth and oval-shaped spores (3–10 m). The strain grew in all of the International Streptomyces Project (ISP)’s media at 28–37 °C with a pH of 6–9 and 0–10% NaCl. S. pakalii ENCB-J15 assimilated diverse carbon as well as organic and inorganic nitrogen sources. The strain also exhibited significant inhibitory activity against the prodigiosin synthesis of Serratia marcescens and the inhibition of the formation and destruction of biofilms of ESKAPE strains of Acinetobacter baumannii and Klebsiella pneumoniae. The draft genome sequencing of ENCB-J15 revealed a 7.6 Mb genome with a high G + C content (71.6%), 6833 total genes, and 6746 genes encoding putative proteins. A total of 26 accessory clusters of proteins associated with carbon sources and amino acid catabolism, DNA modification, and the antibiotic biosynthetic process were annotated. The 16S rRNA gene phylogeny, core-proteome phylogenomic tree, and virtual genome fingerprints support that S. pakalii ENCB-J15 is a new species related to Streptomyces badius and Streptomyces globisporus. Similarly, its average nucleotide identity (ANI) (96.4%), average amino acid identity (AAI) (96.06%), and virtual DNA–DNA hybridization (67.3%) provide evidence to recognize it as a new species. Comparative genomics revealed that S. pakalli and its closest related species maintain a well-conserved genomic synteny. This work proposes Streptomyces pakalii sp. nov. as a novel species that expresses anti-biofilm and anti-quorum sensing activities. Full article

The main aim of this study is to find relevant analytic fingerprints for plants’ structural characterization using spectroscopic techniques and thermogravimetric analyses (TGAs) as alternative methods, particularized on cabbage treated with selenium–baker’s yeast vinasse formulation (Se-VF) included in a foliar fertilizer formula. The hypothesis investigated is that Se-VF will induce significant structural changes compared with the control, analytically confirming the biofortification of selenium-enriched cabbage as a nutritive vegetable, and particularly the plant biostimulant effects of the applied Se-VF formulation on cabbage grown in the field. The TGA evidenced a structural transformation of the molecular building blocks in the treated cabbage leaves. The ash residues increased after treatment, suggesting increased mineral accumulation in leaves. X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR) evidenced a pectin–Iα-cellulose structure of cabbage that correlated with each other in terms of leaf crystallinity. FTIR analysis suggested the accumulation of unesterified pectin and possibly (seleno) glucosinolates and an increased network of hydrogen bonds. The treatment with Se-VF formulation induced a significant increase in the soluble fibers of the inner leaves, accompanied by a decrease in the insoluble fibers. The ratio of soluble/insoluble fibers correlated with the crystallinity determined by XRD and with the FTIR data. The employed analytic techniques can find practical applications as fast methods in studies of the effects of new agrotechnical practices, while in our particular case study, they revealed effects specific to plant biostimulants of the Se-VF formulation treatment: enhanced mineral utilization and improved quality traits. Full article

Coffea liberica presents the smallest proportion of cultivated coffee worldwide. Its taste varies highly from C. arabica spp. or C. canephora spp. As the unfermented taste of C. liberica is often described as lactic, and animalic with aromatics of blue cheese, fermentation experiments of C. liberica coffee cherries were conducted. Coffee cherry fermentation is carried out to modify and enhance the sensory profile of green and roasted coffee beans. Various microorganisms change the composition of organic precursor compounds, reducing off-flavours during green bean coffee processing and roasting. For the comparison of the sensory properties of fermented coffee beans, SCA-trained experts grade the coffees by specific sensory attributes. Roasted and ground coffee powder, as well as ground raw coffee samples, were analysed by headspace-gas chromatography–mass spectrometry/ion mobility spectrometry (GC-MS/IMS) to determine the volatile fingerprint of unfermented and fermented C. liberica coffee. Eight different unfermented C. liberica samples were analysed and the volatile profiles compared to those of arabica spp. and canephora spp. The unfermented coffee beans differ in origin and processing. Furthermore, single-origin C. liberica fermented with two different wine yeasts was compared to unfermented C. liberica. The fermented coffee beans share the same variety, processing, and profile of roasting and differ only in their origin and fermentation inoculum. This study presents the direct comparison of the aromatic fingerprint measured in the instrument. Compound regions in the spectra associated with different coffee species and ferments are shown. The potential of GC-IMS for fast comparison of the aromatic fingerprint is demonstrated. Full article

Given the substantial world coffee production, tons of coffee fruit cascara rich in bioactive compounds are discarded annually. Using this by-product to produce potentially healthy and acceptable foods is a sustainable practice that aggregates value to coffee production and may help improve people’s lives. This study aimed to elaborate kombuchas from coffee cascara tea, evaluate their microbial profile, and monitor the changes in the volatile profile during fermentation, together with sensory attributes and acceptance by consumers from Rio de Janeiro (n = 113). Arabica coffee cascaras from Brazil and Nicaragua were used to make infusions, to which black tea kombucha, a Symbiotic Culture of Bacteria and Yeasts (SCOBY), and sucrose were added. Fermentation of plain black tea kombucha was also monitored for comparison. The volatile profile was analyzed after 0, 3, 6, and 9 days of fermentation via headspace solid phase microextraction GC-MS. A total of 81 compounds were identified considering all beverages, 59 in coffee cascara kombuchas and 59 in the black tea kombucha, with 37 common compounds for both. An increase mainly in acids and esters occurred during fermentation. Despite the similarity to black tea kombucha, some aldehydes, esters, alcohols, and ketones in coffee cascara kombucha were not identified in black tea kombucha. Potential impact compounds in CC were linalool, decanal, nonanal, octanal, dodecanal, ethanol, 2-ethylhexanol, ethyl acetate, ethyl butyrate, ethyl acetate, β-damascenone, γ-nonalactone, linalool oxide, phenylethyl alcohol, geranyl acetone, phenylacetaldehyde, isoamyl alcohol, acetic acid, octanoic acid, isovaleric acid, ethyl isobutyrate, ethyl hexanoate, and limonene. The mean acceptance scores for cascara kombuchas varied between 5.7 ± 0.53 and 7.4 ± 0.53 on a nine-point hedonic scale, with coffee cascara from three-day Nicaragua kombucha showing the highest score, associated with sweetness and berry, honey, woody, and herbal aromas and flavors. The present results indicate that coffee cascara is a promising by-product for elaboration of fermented beverages, exhibiting exotic and singular fingerprinting that can be explored for applications in the food industry. Full article