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1.
Sci Rep ; 14(1): 14726, 2024 06 26.
Article in English | MEDLINE | ID: mdl-38926524

ABSTRACT

Zoonotic yeast species have been implicated in disease development in both humans and cats. This study analyzed the yeast mycobiota present in feline facial hair and human nails and explored potential interspecies associations. A total of 118 biological specimens were examined, including 59 feline facial hair and 59 human nail samples. DNA extraction and DNA sequencing were performed to identify the specific yeast species. The most predominant yeast species in humans and cats were selected for antifungal susceptibility testing (itraconazole, ketoconazole, miconazole, and terbinafine). The findings unveiled diverse yeast species in cats and humans. Malassezia pachydermatis (45.8%) and Malassezia furfur (30.5%) were the most common yeast species in cats and humans, respectively. However, no significant correlation was detected between the yeast species identified in cats and their owners residing in the same household (p > 0.05). Miconazole exhibited the highest minimum inhibitory concentrations (MICs) against Malassezia pachydermatis and Malassezia furfur in both cat and human isolates, whereas terbinafine showed the lowest MICs against most Malassezia pachydermatis and Malassezia furfur in both cat and human isolates. Diverse yeast species in cat facial hair and human nails suggest possible cross-contamination among humans, pets, and environments.


Subject(s)
Antifungal Agents , Microbial Sensitivity Tests , Nails , Cats , Humans , Antifungal Agents/pharmacology , Animals , Nails/microbiology , Malassezia/drug effects , Malassezia/genetics , Malassezia/isolation & purification , Hair/microbiology , Yeasts/drug effects , Yeasts/isolation & purification , Yeasts/genetics , Terbinafine/pharmacology , Miconazole/pharmacology , Male , Animal Fur/microbiology , Female
2.
Food Res Int ; 187: 114366, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38763646

ABSTRACT

In recent years, numerous studies have demonstrated the significant potential of non-Saccharomyces yeasts in aroma generation during fermentation. In this study, 134 strains of yeast were isolated from traditional fermented foods. Subsequently, through primary and tertiary screening, 28 strains of aroma-producing non-Saccharomyces yeast were selected for beer brewing. Headspace-solid phase microextraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS) and chemometrics were employed to analyze the volatile flavor substances in beer samples fermented using these strains. Chemometric analysis revealed that distinct species of non-Saccharomyces yeast had a unique influence on beer aroma, with strains from the same genus producing more similar flavor profiles. Accordingly, 2,6-nonadienal, 1-pentanol, phenyl ethanol, isoamyl acetate, ethyl caprate, butyl butyrate, ethyl propionate, furfuryl alcohol, phenethyl acetate, ethyl butyrate, ethyl laurate, acetic acid, and 3-methyl-4 heptanone were identified as the key aroma compounds for distinguishing among different non-Saccharomyces yeast species. This work provides useful insights into the aroma-producing characteristics of different non-Saccharomyces yeasts to reference the targeted improvement of beer aroma.


Subject(s)
Beer , Fermentation , Fermented Foods , Gas Chromatography-Mass Spectrometry , Odorants , Solid Phase Microextraction , Volatile Organic Compounds , Yeasts , Beer/analysis , Beer/microbiology , Odorants/analysis , Volatile Organic Compounds/analysis , Fermented Foods/microbiology , Fermented Foods/analysis , Yeasts/isolation & purification , Yeasts/metabolism , Food Microbiology
3.
BMC Microbiol ; 24(1): 163, 2024 May 14.
Article in English | MEDLINE | ID: mdl-38745280

ABSTRACT

Spontaneous fermentation of cereals like millet involves a diverse population of microbes from various sources, including raw materials, processing equipment, fermenting receptacles, and the environment. Here, we present data on the predominant microbial species and their succession at each stage of the Hausa koko production process from five regions of Ghana. The isolates were enumerated using selective media, purified, and phenotypically characterised. The LAB isolates were further characterised by 16S rRNA Sanger sequencing, typed using (GTG)5 repetitive-PCR, and whole genome sequencing, while 28S rRNA Sanger sequencing was performed for yeast identification. The pH of the millet grains ranged from mean values of 6.02-6.53 to 3.51-3.99 in the final product, depending on the processors. The mean LAB and yeast counts increased during fermentation then fell to final counts of log 2.77-3.95 CFU/g for LAB and log 2.10-2.98 CFU/g for yeast in Hausa koko samples. At the various processing stages, the counts of LAB and yeast revealed significant variations (p < 0.0001). The species of LAB identified in this study were Limosilactobacillus pontis, Pediococcus acidilactici, Limosilactobacillus fermentum, Limosilactobacillus reuteri, Pediococcus pentosaceus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Schleiferilactobacillus harbinensis, and Weissella confusa. The yeasts were Saccharomyces cf. cerevisiae/paradoxus, Saccharomyces cerevisiae, Pichia kudriavzevii, Clavispora lusitaniae and Candida tropicalis. The identification and sequencing of these novel isolates and how they change during the fermentation process will pave the way for future controlled fermentation, safer starter cultures, and identifying optimal stages for starter culture addition or nutritional interventions. These LAB and yeast species are linked to many indigenous African fermented foods, potentially acting as probiotics in some cases. This result serves as the basis for further studies into the technological and probiotic potential of these Hausa koko microorganisms.


Subject(s)
Fermentation , Fermented Foods , Food Microbiology , Millets , Yeasts , Ghana , Yeasts/classification , Yeasts/isolation & purification , Yeasts/genetics , Yeasts/metabolism , Fermented Foods/microbiology , Millets/microbiology , Lactobacillales/classification , Lactobacillales/isolation & purification , Lactobacillales/genetics , Lactobacillales/metabolism , RNA, Ribosomal, 16S/genetics , Phylogeny , Hydrogen-Ion Concentration , Edible Grain/microbiology
4.
FEMS Microbiol Lett ; 3712024 Jan 09.
Article in English | MEDLINE | ID: mdl-38782713

ABSTRACT

Recent research has shown the potential of yeast-based biosensors (YBBs) for point-of-use detection of pathogens and target molecules in saliva, blood, and urine samples. The choice of output can greatly affect the sensitivity, dynamic range, detection time, and ease-of-use of a sensor. For visual detection without the need for additional reagents or machinery, colorimetric outputs have shown great potential. Here, we evaluated the inducible generation of prodeoxyviolacein and proviolacein as colorimetric YBB outputs and benchmarked these against lycopene. The outputs were induced via the yeast mating pathway and were compared on agar plates, in liquid culture, and on paper slips. We found that all three outputs produced comparable pigment intensity on agar plates, making them applicable for bioengineering settings. In liquid media and on paper slips, lycopene resulted in a higher intensity pigment and a decreased time-of-detection.


Subject(s)
Biosensing Techniques , Colorimetry , Saccharomyces cerevisiae , Biosensing Techniques/methods , Colorimetry/methods , Saccharomyces cerevisiae/metabolism , Lycopene/metabolism , Yeasts/isolation & purification , Yeasts/metabolism , Carotenoids/analysis , Carotenoids/metabolism , Point-of-Care Systems
5.
J Am Soc Mass Spectrom ; 35(6): 1352-1362, 2024 Jun 05.
Article in English | MEDLINE | ID: mdl-38742647

ABSTRACT

Mass spectrometry is commonly used in the identification of species present in microbial samples, but the high similarity in the peptide composition between strains of a single species has made analysis at the subspecies level challenging. Prior research in this area has employed methods such as Principal Component Analysis (PCA), the k-Nearest Neighbors' (kNN) algorithm, and Pearson correlation. Previously, 1D cross-correlation of mass spectra has been shown to be useful in the classification of small molecule compounds as well as in the identification of peptide sequences via the SEQUEST algorithm and its variants. While direct application of cross-correlation to mass spectral data has been shown to aid in the identification of many other types of compounds, this type of analysis has not been demonstrated in the literature for the purpose of LC-MS based identification of microbial strains. A method of identifying microbial strains is presented here that applies the principle of 2D cross-correlation to LC-MS data. For a set of N = 30 yeast isolate samples representing 5 yeast strains (K-97, S-33, T-58, US-05, WB-06), high-resolution LC-MS-Orbitrap data were collected. Reference spectra were then generated for each strain from the combined data of each sample of that strain. Sample strains were then predicted by computing the 2D cross-correlation of each sample against the reference spectra, followed by application of correction factors measuring the asymmetry of the 2D correlation functions.


Subject(s)
Algorithms , Mass Spectrometry , Chromatography, Liquid/methods , Mass Spectrometry/methods , Yeasts/chemistry , Yeasts/classification , Yeasts/isolation & purification , Liquid Chromatography-Mass Spectrometry
6.
Environ Microbiol Rep ; 16(3): e13213, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38738810

ABSTRACT

Since a significant proportion of plant matter is consumed by herbivores, a necessary adaptation for many phyllosphere microbes could be to survive through the guts of herbivores. While many studies explore the gut microbiome of herbivores by surveying the microbiome in their frass, few studies compare the phyllosphere microbiome to the gut microbiome of herbivores. High-throughput metabarcode sequencing was used to track the fungal community from milkweed (Asclepias spp.) leaves to monarch caterpillar frass. The most commonly identified fungal taxa that dominated the caterpillar frass after the consumption of leaves were yeasts, mostly belonging to the Basidiomycota phylum. While most fungal communities underwent significant bottlenecks and some yeast taxa increased in relative abundance, a consistent directional change in community structure was not identified from leaf to caterpillar frass. These results suggest that some phyllosphere fungi, especially diverse yeasts, can survive herbivory, but whether herbivory is a key stage of their life cycle remains uncertain. For exploring phyllosphere fungi and the potential coprophilous lifestyles of endophytic and epiphytic fungi, methods that target yeast and Basidiomycota fungi are recommended.


Subject(s)
Asclepias , Fungi , Herbivory , Plant Leaves , Animals , Plant Leaves/microbiology , Asclepias/microbiology , Fungi/classification , Fungi/genetics , Fungi/isolation & purification , Fungi/physiology , Yeasts/classification , Yeasts/isolation & purification , Yeasts/genetics , Mycobiome , Basidiomycota/classification , Basidiomycota/genetics , Basidiomycota/physiology , Basidiomycota/isolation & purification , Gastrointestinal Microbiome , Larva/microbiology , Moths/microbiology
7.
Med Mycol ; 62(5)2024 May 03.
Article in English | MEDLINE | ID: mdl-38627248

ABSTRACT

Although Candida species are the most common cause of fungemia, non-Candida rare yeasts (NCY) have been increasingly reported worldwide. Although the importance of these yeast infections is recognized, current epidemiological information about these pathogens is limited, and they have variable antifungal susceptibility profiles. In this study, we aimed to evaluate the clinical characteristics for fungemia caused by NCY by comparing with candidemia. The episodes of NCY fungemia between January 2011 and August 2023 were retrospectively evaluated in terms of clinical characteristics, predisposing factor, and outcome. In addition, a candidemia group, including patients in the same period was conducted for comparison. Antifungal susceptibility tests were performed according to the reference method. A total of 85 patients with fungemia episodes were included: 25 with NCY fungemia and 60 with candidemia. Fluconazole had high minimal inhibitory concentration (MIC) values against almost all NCY isolates. The MIC values for voriconazole, posaconazole, and amphotericin B were ≤ 2 µg/ml, and for caspofungin and anidulafungin were ≥ 1 µg/ml against most of isolates. Hematological malignancies, immunosuppressive therapy, neutropenia and prolonged neutropenia, polymicrobial bacteremia/fungemia, preexposure to antifungal drugs, and breakthrough fungemia were associated with NCY fungemia, whereas intensive care unit admission, diabetes mellitus, urinary catheters, and total parenteral nutrition were associated with candidemia. In conclusion, the majority of fungemia due to NCY species was the problem, particularly in hematology units and patients with hematological malignancy. Preexposure to antifungal drugs likely causes a change in the epidemiology of fungemia in favor of non-albicans Candida and/or NCY.


Among all fungemia episodes, hematological malignancies, immunosuppressive therapy, neutropenia, and preexposure to antifungals were risk factors for non-Candida yeast fungemia; diabetes mellitus, urinary catheters, and total parenteral nutrition were risks for candidemia.


Subject(s)
Antifungal Agents , Candida , Candidemia , Fungemia , Microbial Sensitivity Tests , Tertiary Care Centers , Humans , Retrospective Studies , Tertiary Care Centers/statistics & numerical data , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Male , Female , Middle Aged , Aged , Candida/drug effects , Candida/isolation & purification , Candida/classification , Fungemia/microbiology , Fungemia/epidemiology , Fungemia/drug therapy , Adult , Candidemia/microbiology , Candidemia/epidemiology , Candidemia/drug therapy , Yeasts/isolation & purification , Yeasts/drug effects , Yeasts/classification , Aged, 80 and over , Fluconazole/pharmacology , Fluconazole/therapeutic use , Young Adult
8.
Braz J Microbiol ; 55(2): 1451-1463, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38656427

ABSTRACT

Antarctic temperature variations and long periods of freezing shaped the evolution of microorganisms with unique survival mechanisms. These resilient organisms exhibit several adaptations for life in extreme cold. In such ecosystems, microorganisms endure the absence of liquid water and exhibit resistance to freezing by producing water-binding molecules such as antifreeze proteins (AFP). AFPs modify the ice structure, lower the freezing point, and inhibit recrystallization. The objective of this study was to select and identify microorganisms isolated from different Antarctic ecosystems based on their resistance to temperatures below 0 °C. Furthermore, the study sought to characterize these microorganisms regarding their potential antifreeze adaptive mechanisms. Samples of soil, moss, permafrost, and marine sediment were collected on King George Island, located in the South Shetland archipelago, Antarctica. Bacteria and yeasts were isolated and subjected to freezing-resistance and ice recrystallization inhibition (IR) tests. A total of 215 microorganisms were isolated, out of which 118 were molecularly identified through molecular analysis using the 16S rRNA and ITS regions. Furthermore, our study identified 24 freezing-resistant isolates, including two yeasts and 22 bacteria. A total of 131 protein extracts were subjected to the IR test, revealing 14 isolates positive for AFP production. Finally, four isolates showed both freeze-resistance and IR activity (Arthrobacter sp. BGS04, Pseudomonas sp. BGS05, Cryobacterium sp. P64, and Acinetobacter sp. M1_25C). This study emphasizes the diversity of Antarctic microorganisms with the ability to tolerate freezing conditions. These microorganisms warrant further investigation to conduct a comprehensive analysis of their antifreeze capabilities, with the goal of exploring their potential for future biotechnological applications.


Subject(s)
Antifreeze Proteins , Bacteria , Freezing , Antarctic Regions , Antifreeze Proteins/metabolism , Antifreeze Proteins/chemistry , Antifreeze Proteins/genetics , Bacteria/genetics , Bacteria/classification , Bacteria/metabolism , Bacteria/isolation & purification , Islands , Phylogeny , Yeasts/genetics , Yeasts/classification , Yeasts/isolation & purification , Yeasts/metabolism , RNA, Ribosomal, 16S/genetics , Ecosystem
9.
Int J Food Microbiol ; 417: 110688, 2024 Jun 02.
Article in English | MEDLINE | ID: mdl-38615425

ABSTRACT

Taggiasca table olives are typical of Liguria, a Northwestern Italian region, produced with a spontaneous fermentation carried out by placing the raw drupes directly into brine with a salt concentration of 8-12 % w/v. Such concentrations limit the development of unwanted microbes and favor the growth of yeasts. This process usually lasts up to 8 months. Yeasts are found throughout the entire fermentation process and they are mainly involved in the production of volatile organic compounds, which strongly impact the quality of the final product. The aim of this study was to evaluate the dynamics of autochthonous yeasts in brines and olives in a spontaneous process with no lye pre-treatment or addition of acids in the fermenting brine with 10 % NaCl (w/v) in two batches during 2021 harvest. Three hundred seventy-three yeast colonies were isolated, characterized by rep-PCR and identified by the D1/D2 region of the 26S rRNA gene sequencing. Mycobiota was also studied by 26S rRNA gene metataxonomics, while metabolome was assessed through GC-MS analysis. Traditional culture-dependent methods showed the dominance of Candida diddensiae, Wickerhamomyces anomalus, Pichia membranifaciens and Aureobasidium pullulans, with differences in species distribution between batches, sampling time and type of sample (olives/brines). Amplicon-based sequencing confirmed the dominance of W. anomalus in batch 1 throughout the entire fermentation, while Cyteromyces nyonsensis and Aureobasidium spp. were most abundant in the fermentation in batch 2. Volatilome results were analyzed and correlated to the mycobiota data, confirming differences between fermentation stages. Given the high appreciation for this traditional food, this study helps elucidate the mycobiota associated to Taggiasca cv. table olives and its relationship with the quality of the final product.


Subject(s)
Fermentation , Food Microbiology , Olea , Volatile Organic Compounds , Yeasts , Olea/microbiology , Volatile Organic Compounds/analysis , Volatile Organic Compounds/metabolism , Yeasts/metabolism , Yeasts/classification , Yeasts/isolation & purification , Yeasts/genetics , Italy , Salts
10.
J Appl Microbiol ; 135(5)2024 May 01.
Article in English | MEDLINE | ID: mdl-38621715

ABSTRACT

AIMS: To compare the species diversity and composition of indigenous yeast communities of hybrid grapes from conventionally and organically cultivated vineyards of an emerging cool-climate wine producing region. METHODS AND RESULTS: Illumina MiSeq sequences from L'Acadie blanc grape musts were processed and filtered to characterize indigenous yeast communities in organic and conventional vineyards of the Annapolis Valley wine region in Nova Scotia, Canada. While cultivation practice was not associated with yeast diversity or species richness, there was a strong effect on yeast community composition, with conventional vineyards characterized by higher proportions of Sporidiobolales and Filobasidium magnum, and organic vineyards supporting Filobasidium species other than F. magnum and higher proportions of Symmetrospora. There was also variation in yeast community composition among individual vineyards, and from year to year. CONCLUSIONS: This is the first comprehensive assessment of yeasts associated with hybrid grapes grown using different cultivation practices in a North American cool climate wine region. Communities were dominated by basidiomycete yeasts and species composition of these yeasts differed significantly between vineyards employing organic and conventional cultivation practices. The role of basidiomycete yeasts in winemaking is not well understood, but some species may influence wine characteristics.


Subject(s)
Vitis , Wine , Yeasts , Vitis/microbiology , Wine/microbiology , Wine/analysis , Yeasts/genetics , Yeasts/classification , Yeasts/isolation & purification , Nova Scotia , Farms , Organic Agriculture
11.
Diagn Microbiol Infect Dis ; 109(2): 116270, 2024 Jun.
Article in English | MEDLINE | ID: mdl-38518388

ABSTRACT

MALDI-TOF MS identifications of microorganisms in a clinical laboratory were investigated, comparing steel targets with MBT Biotargets. By using MBT Biotargets, the score values of yeast identifications increased, whereas the score values of Gram-negative bacteria decreased. Switching to MBT Biotargets did not negatively impact overall frequencies of high confidence identifications.


Subject(s)
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Steel , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Humans , Steel/chemistry , Bacteria/classification , Bacteria/isolation & purification , Bacteria/genetics , Microbiological Techniques/methods , Yeasts/isolation & purification , Yeasts/classification , Yeasts/genetics
12.
Biol Trace Elem Res ; 201(6): 3069-3076, 2023 Jun.
Article in English | MEDLINE | ID: mdl-35915280

ABSTRACT

The Çökelek samples what 30 different were collected from randomly local bazaars to investigate heavy metal contaminant and mineral levels and some physicochemical and microbiological properties of samples. While the Pb was identified in 6 of the 30 samples, the As was only found in 4 of the samples. The mean major and trace element contents of Çökelek samples were ordered as Na > P > Ca > K > Mg and Al > Zn > Ni > Cu, respectively. The physicochemical properties indicated a high deviation among samples. The mean total solids, ash, salt, fat, protein waters soluble nitrogen contents, and sample ripening index were 29.83%, 1.88%, 0.68%, 4.31%, 19.84%, 0.33%, and 1.79%, respectively. The mean total aerobic mesophilic bacteria (TAMB) count of Çökelek samples was found as 8.26 log CFU g-1. The coliform bacteria and yeast-mold counts were detected in 11 and 27 of 30 samples, respectively. The mean coliform and yeast-mold counts were 1.82 log CFU g-1 and 7.11 log CFU g-1, respectively. Traditional cheeses are not mentioned in legal laws such as the Turkish Food Codex. So, there is no legal limit and standard production processes. This situation is a problem in terms of traditional cheese quality. For this reason, traditional cheese should perform further studied, and determine the legal limits.


Subject(s)
Cheese , Food Microbiology , Cheese/analysis , Cheese/microbiology , Food Handling , Fungi/isolation & purification , Sodium Chloride , Turkey , Yeasts/isolation & purification , Colony Count, Microbial , Enterobacteriaceae/isolation & purification
13.
Molecules ; 27(3)2022 Jan 28.
Article in English | MEDLINE | ID: mdl-35164165

ABSTRACT

Yeasts are one of the main ingredients responsible for flavor precursors production associated with sensorial characteristics in chocolate. Using wild yeast isolated from cocoa beans fermentation is emerging as a strategy for developing starter cultures. However, the volatile compounds (VCs) produced by yeasts are not yet known. This study aimed to select wild yeasts with the potential to produce volatile compounds associated with desirable flavor attributes. A total of 150 wild yeasts strains were isolated from the spontaneous cocoa beans fermentation, of which 40 were identified by morphology and physiological features. VCs produced were identified and quantified using SPME-GC-MS and GC-FID and profiles were evaluated statistically by PCA and cluster analysis for the compounds that had a high odor threshold value. Thirty-six VCs produced by these yeasts were identified into six main families, namely esters, alcohols, acids, aldehydes, ketones, and pyrazines. PCA showed the separation of the yeasts into two main clusters. Strains, Y195 and Y246, belong to the first cluster and are the highest producers of alcohols related to floral perceptions. In the second cluster, thirty-three yeasts were grouped by their ability to produce esters. Of all of them, Y110MRS stood out for producing 2-phenyl ethyl acetate and isoamyl acetate associated with fruity perceptions. This screening allowed us to identify yeasts that produced VCs of technological interest and which could be used to develop a starter culture.


Subject(s)
Cacao/microbiology , Fermentation , Volatile Organic Compounds/analysis , Yeasts/isolation & purification , Alcohols/analysis , Aldehydes/analysis , Cacao/metabolism , Chocolate/analysis , Pyrazines/analysis , Yeasts/metabolism
15.
J Clin Microbiol ; 60(1): e0139821, 2022 01 19.
Article in English | MEDLINE | ID: mdl-34669460

ABSTRACT

Fourier transform infrared (FTIR) spectroscopy has demonstrated applicability as a reagent-free whole-organism fingerprinting technique for both microbial identification and strain typing. For routine application of this technique in microbiology laboratories, acquisition of FTIR spectra in the attenuated total reflectance (ATR) mode simplifies the FTIR spectroscopy workflow, providing results within minutes after initial culture without prior sample preparation. In our previous central work, 99.7% correct species identification of clinically relevant yeasts was achieved by employing an ATR-FTIR-based method and spectral database developed by our group. In this study, ATR-FTIR spectrometers were placed in 6 clinical microbiology laboratories over a 16-month period and were used to collect spectra of routine yeast isolates for on-site identification to the species level. The identification results were compared to those obtained from conventional biochemical tests and/or matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Isolates producing discordant results were reanalyzed by routine identification methods, ATR-FTIR spectroscopy, and PCR gene sequencing of the D1/D2 and internal transcribed spacer (ITS) regions. Among the 573 routine clinical yeast isolates collected and identified by the ATR-FTIR-based method, 564 isolates (98.4%) were correctly identified at the species level, while the remaining isolates were inconclusive with no misidentifications. Due to the low prevalence of Candida auris in routine isolates, additional randomly selected C. auris (n = 24) isolates were obtained for evaluation and resulted in 100% correct identification. Overall, the data obtained in our multicenter evaluation study using multiple spectrometers and system operators indicate that ATR-FTIR spectroscopy is a reliable, cost-effective yeast identification technique that provides accurate and timely (∼3 min/sample) species identification promptly after the initial culture.


Subject(s)
Yeasts , Fourier Analysis , Humans , Spectroscopy, Fourier Transform Infrared/methods , Yeasts/isolation & purification
16.
Microbiol Spectr ; 9(3): e0140421, 2021 12 22.
Article in English | MEDLINE | ID: mdl-34935420

ABSTRACT

In the beer brewing industry, microbial spoilage presents a consistent threat that must be monitored and controlled to ensure the palatability of a finished product. Many of the predominant beer spoilage microbes have been identified and characterized, but the mechanisms of contamination and persistence remain an open area of study. Postproduction, many beers are distributed as kegs that are attached to draft delivery systems in retail settings where ample opportunities for microbial spoilage are present. As such, restaurants and bars can experience substantial costs and downtime for cleaning when beer draft lines become heavily contaminated. Spoilage monitoring on the retail side of the beer industry is often overlooked, yet this arena may represent one of the largest threats to the profitability of a beer if its flavor profile becomes substantially distorted by contaminating microbes. In this study, we sampled and cultured microbial communities found in beers dispensed from a retail draft system to identify the contaminating bacteria and yeasts. We also evaluated their capability to establish new biofilms in a controlled setting. Among four tested beer types, we identified over a hundred different contaminant bacteria and nearly 20 wild yeasts. The culturing experiments demonstrated that most of these microbes were viable and capable of joining new biofilm communities. These data provide an important reference for monitoring specific beer spoilage microbes in draft systems and we provide suggestions for cleaning protocol improvements. IMPORTANCE Beer production, packaging, and service are each vulnerable to contamination by microbes that metabolize beer chemicals and impart undesirable flavors, which can result in the disposal of entire batches. Therefore, great effort is taken by brewmasters to reduce and monitor contamination during production and packaging. A commonly overlooked quality control stage of a beer supply chain is at the retail service end, where beer kegs supply draft lines in bars and restaurants under nonsterile conditions. We found that retail draft line contamination is rampant and that routine line cleaning methods are insufficient to efficiently suppress beer spoilage. Thus, many customers unknowingly consume spoiled versions of the beers they consume. This study identified the bacteria and yeast that were resident in retail draft beer samples and also investigated their abilities to colonize tubing material as members of biofilm communities.


Subject(s)
Bacterial Physiological Phenomena , Beer/microbiology , Biofilms , Microbiota , Yeasts/physiology , Bacteria/classification , Bacteria/genetics , Bacteria/isolation & purification , Food Contamination/analysis , Food Microbiology , Yeasts/classification , Yeasts/genetics , Yeasts/isolation & purification
17.
FEMS Microbiol Lett ; 368(20)2021 11 27.
Article in English | MEDLINE | ID: mdl-34755861

ABSTRACT

Non-Saccharomyces yeasts are important players during winemaking and may come from grapes grown in vineyards. To study the diversity of non-Saccharomyces yeasts on grape berry surfaces, 433 strains were isolated from different Cabernet Sauvignon vineyards grown in Henan Province. Our results demonstrated that these strains were classified into 16 morphotypes according to their growth morphology on Wallerstein Laboratory agar medium, and were identified as seven species from four genera-Hanseniaspora opuntiae, Hanseniaspora vineae, Hanseniaspora uvarum, Pichia occidentalis, Pichia kluyveri, Issatchenkia terricola and Saturnispora diversa-based on a series of molecular biological experiments. Hanseniaspora opuntiae was obtained from all sampling sites except Changyuan County, while Pichia kluyveri and Saturnispora diversa were only found in sites of Zhengzhou Grape Resource Garden and Minquan County, respectively. The site Minquan was home of the greatest species richness, while only one single species (Hanseniaspora opuntiae) was detected at NAPA winery from Zhengzhou or at Anyang County. Finally, this study suggested that the geographic distribution and diversity of non-Saccharomyces yeast populations on Cabernet Sauvignon grape berries were likely to be determined by a combination of grape varieties and environmental factors.


Subject(s)
Biodiversity , Fruit , Vitis , Yeasts , China , Farms , Fermentation , Fruit/microbiology , Hanseniaspora/classification , Hanseniaspora/isolation & purification , Pichia/classification , Pichia/isolation & purification , Saccharomycetales/classification , Saccharomycetales/isolation & purification , Vitis/microbiology , Wine/microbiology , Yeasts/classification , Yeasts/isolation & purification
18.
FEMS Microbiol Lett ; 368(20)2021 11 30.
Article in English | MEDLINE | ID: mdl-34788824

ABSTRACT

Nine morphologically distinct halophilic yeasts were isolated from Makgadikgadi and Sua pans, as pristine and extreme environments in Botswana. Screening for biosurfactant production showed that Rhodotorula mucilaginosa SP6 and Debaryomyces hansenii MK9 exhibited the highest biosurfactant activity using Xanthocercis zambesiaca seed powder as a novel and alternative inexpensive carbon substrate. Chemical characterization of the purified biosurfactants by Fourier Transform Infra-Red spectroscopy suggested that the biosurfactant from R. mucilaginosa SP6 was a rhamnolipid-type whereas the biosurfactant from D. hansenii MK9 was a sophorolipid-type. The two biosurfactants exhibited antimicrobial activities against eight pathogenic bacteria and fungal strains (Proteus vulgaris, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Micrococcus luteus, Cryptococcus neoformans, Candida albicans and Aspergilus niger). The sophorolopid-type biosurfactant was found to be the most potent among the antimicrobial drug resistant strains tested. The findings open up prospects for the development of environmentally friendly antimicrobial drugs that use an inexpensive source of carbon to reduce the costs associated with the production of biosurfactants.


Subject(s)
Extreme Environments , Surface-Active Agents , Yeasts , Anti-Infective Agents/isolation & purification , Anti-Infective Agents/metabolism , Anti-Infective Agents/pharmacology , Bacteria/drug effects , Botswana , Carbon/metabolism , Debaryomyces/chemistry , Debaryomyces/metabolism , Fungi/drug effects , Industrial Microbiology , Rhodotorula/chemistry , Rhodotorula/metabolism , Surface-Active Agents/isolation & purification , Surface-Active Agents/metabolism , Surface-Active Agents/pharmacology , Yeasts/chemistry , Yeasts/isolation & purification , Yeasts/metabolism
19.
Biol Futur ; 72(3): 325-337, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34554551

ABSTRACT

Plasmodial resistance to a variety of plant-based antimalarial drugs has led toward the discovery of more effective antimalarial compounds having chemical or biological origin. Since natural compounds are considered as safer drugs, in this study, yeast strains were identified and compared for the production of carotenoids that are well-known antioxidants and this metabolite was tested for its antiparasitic activity. Plasmodium falciparum 3D7 strain was selected as the target parasite for evaluation of antimalarial activity of yeast carotenoids using in vitro studies. Data were analyzed by FACS (fluorescence-activated cell sorter) and counted via gold standard Giemsa-stained smears. The extracted yeast carotenoids showed a profound inhibitory effect at a concentration of 10-3 µg/µl and 10-4 µg/µl when compared to ß- carotene as control. SYBR Green1 fluorescent dye was used to confirm the decrease in parasitaemia at given range of concentration. Egress assay results suggested that treated parasite remained stalled at schizont stage with constricted morphology and were darkly stained. Non-toxicity of carotenoids on erythrocytes and on human liver hepatocellular carcinoma cells (HepG2 cells) was shown at a given concentration. This report provides strong evidence for antimalarial effects of extracted yeast carotenoids, which can be produced via a sustainable and cost-effective strategy and may be scaled up for industrial application.


Subject(s)
Antimalarials/standards , Carotenoids/analysis , Carotenoids/isolation & purification , Plasmodium falciparum/drug effects , Yeasts/metabolism , Antimalarials/pharmacology , Antimalarials/therapeutic use , Malaria, Falciparum/drug therapy , Malaria, Falciparum/physiopathology , Yeasts/isolation & purification
20.
Food Microbiol ; 100: 103864, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34416964

ABSTRACT

While the trend in winemaking is toward reducing the inputs and especially sulphites utilization, emerging technologies for the preservation of wine is a relevant topic for the industry. Amongst yeast spoilage in wine, Brettanomyces bruxellensis is undoubtedly the most feared. In this study, UV-C treatment is investigated. This non-thermal technique is widely used for food preservation. A first approach was conducted using a drop-platted system to compare the sensitivity of various strains to UV-C surface treatment. 147 strains distributed amongst fourteen yeast species related to wine environment were assessed for six UV-C doses. An important variability in UV-C response was observed at the interspecific level. Interestingly, cellar resident species, which are mainly associated with wine spoilage, shows higher sensitivity to UV-C than vineyard-resident species. A focus on B. bruxellensis species with 104 screened strains highlighted an important effect of the UV-C, with intra-specific variation. This intra-specific variation was confirmed on 6 strains in liquid red wine by using a home-made pilot. 6624 J.L-1 was enough for a reduction of 5 log10 of magnitude for 5 upon 6 strains. These results highlight the potential of UV-C utilization against wine yeast spoiler at cellar scale.


Subject(s)
Wine/microbiology , Yeasts/radiation effects , Phylogeny , Species Specificity , Ultraviolet Rays , Wine/analysis , Yeasts/genetics , Yeasts/growth & development , Yeasts/isolation & purification
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