Number Of Yeast Cells Research Articles

Effect of Edible Sesame Oil on Growth of Clinical Isolates of Candida albicans

Elderly individuals are at increased risk of oral thrush (oral candidiasis) due to decreased saliva secretion. Due to their antimicrobial properties, edible oils can be effective natural agents for oral care. The objective of the present study was to compare the effects of sesame oil, which is widely used for cooking in Asian countries, and two other edible oils on the growth of both mycelial and yeast forms of five clinical isolates of Candida albicans, a causative microorganism of oral thrush. We assessed the effect of each oil in concentrations of 0.078%, 0.156%, and 0.313% on growth of the mycelial forms of the clinical isolates over 24 hr using the crystal violet method. We also evaluated the effect of each oil on growth of the yeast forms by counting the number of viable yeast cells after culturing in the oils for 24 hr. Sesame oil inhibited the growth of both mycelial and yeast forms. Safflower and olive oil also inhibited the growth of both forms of C. albicans but to a lesser extent than sesame oil. The ability to inhibit the growth of the mycelial form correlated with sesame oil concentration. Roasting influenced growth inhibition ability and high-roasted sesame oil most effectively inhibited the yeast form. The growth inhibitory effect differed among the five isolates. We hypothesize that the sesamin and fatty acid components of sesame oil are involved in its antifungal activity.

Differential Adherence and Expression of Virulence Traits by Candida albicans and Candida parapsilosis in Mono- and Dual-Species Cultures in Artificial Saliva

To evaluate specific virulence factors of Candida albicans and Candida parapsilosis clinical oral isolates in mono- and dual-species culture in the presence of artificial saliva. Two of the strains used in this study were isolated from co-infection (C. albicans AM and C. parapsilosis AM2), and the other two were isolated from single infection (C. albicans AC and C. parapsilosis AD). The number of adhered yeast cells was measured and their enzymatic activity was determined simultaneously. In mono-species culture, C. parapsilosis strains adhered to a higher extent to the surface in comparison with the C. albicans strains. In dual-species culture, the C. parapsilosis strains adhered more in the presence of C. albicans AM. Interestingly, C. albicans AM and C. parapsilosis AD adhered to a higher extent when compared with all other co-cultures. In dual-species culture, the enzymatic activity of C. parapsilosis strains in the presence of C. albicans AC was higher than in the presence of C. albicans AM. The virulence factors of C. albicans and C. parapsilosis differ from strain to strain and are influenced by the presence of other species in culture. To understand the expression of virulence factors in Candida dual-species systems.

Influence of commercial baker's yeasts on bread aroma profiles

Baker's yeast is used for bread fermentation throughout the world and it is very important for the bread quality. The scope of this work was to investigate how the aroma of wheat bread crumb is influenced by yeast fermentation by comparing the aroma formation of bread fermented by seven commercial baker's yeasts. Dough samples were fermented with equal number of yeast cells to equal height and then baked. The volatile components were extracted by dynamic headspace sampling and analyzed by gas chromatography mass spectrometry. The dough fermentation time varied significantly from 40 to 100min. The fermentation compounds 2,3-butanedione and 1-propanol were found in significantly higher concentration in bread fermented with the four baker's yeasts having the shortest fermentation times. Furthermore, 3-methylbutanal, 2-methyl-1-propanol and ethyl acetate were found in significantly higher concentration in two of the yeasts. On the other hand phenylacetaldehyde and 2-phenylethanol were found in significantly higher concentration in bread fermented with two other yeasts. It can be concluded that use of the seven commercial baker's yeasts for bread fermentation resulted in significantly different bread aroma profiles.

Galleria mellonella as a model host to study Paracoccidioides lutzii and Histoplasma capsulatum

Non-mammalian models have been used to investigate fungal virulence. In this work we have explored the use of Galleria mellonella as an infection model for the pathogenic dimorphic fungi Histoplasma capsulatum and Paracoccidioides lutzii. In mammalian models these fungi cause similar infections, and disease outcomes are influenced by the quantity of the infective inocula. We describe a similar aspect in a G. mellonella model and characterize the pathogenesis features in this system. Infection with P. lutzii or H. capsulatum, in all inoculum used, killed larvae at 25 and 37°C. However, there was a lack of correlation between the number of yeast cells used for infection and the time to larvae death, which may indicate that the fungi induce protective responses in a dynamic manner as the lowest concentrations of fungi induced the most rapid death. For both fungi, the degree of larvae melanization was directly proportional to the inocula size, and this effect was visibly more apparent at 37°C. Histological evaluation of the larvae showed a correlation between the inoculum and granuloma-like formation. Our results suggest that G. mellonella is a potentially useful model to study virulence of dimorphic fungi.

황금·황련 추출물의 항균활성이 막걸리 저장성에 미치는 영향

This research was carried out to find herbal preservatives for Makgeolli, as Makgeolli loses its commercial value due to overproduced acidic materials. When Makgeolli was kept at 25℃ to find the changes in acidity, total microbial cell number, yeast cell number, and bacterial species variety, a sudden increase of acidity as well as the disappearance of yeast cells occurred at day 6, and Makgeolli was changed to complete off-flavor. Acetobacter pasteurianus is the main acidifier in Makgeolli and shows a synergy effect in acid formation when cultured in combination with Lactobacillus casei. Among 12 herbs, the ethanol extract of Sutellaria baicalensis showed antimicrobial activity against A. pasteurianus, whereas the ethanol extract of Coptidis rhizoma showed antimicrobial activity against L. casei. Makgeolli added with Sutellaria baicalensis extracts demonstrated a lower acidity than that with Coptidis rhizoma extracts, which indicates that the inhibition of an acetic acid former is more important than that of a lactic acid former in Makgeolli preservation. Sutellaria baicalensis extracts prolonged the shelf life of Makgeolli by 1～2 weeks at a minimal inhibitory concentration (0.63 ㎎/㎖) during storage at 10℃.

젖산농도별 주모에 따른 막걸리의 품질 특성

The quality characteristics of rice wine seed mash and rice wine made with different lactic acid concentrations were investigated. The pH decreased along with the lactic acid concentration. The total titratable acid content of the rice wine seed mash was lowest when 0.5% lactic acid was added, and the ethanol contents of the mash samples were not significant. The results of the measurement of the microorganism number, C (cell numbers of the total bacteria and the lactic acid bacteria), decreased along with the lactic acid concentrations of the mash samples. The yeast cell numbers of the rice wine seed mash samples according to the lactic acid concentrations were high (0.5, 0.3, 1.0, control, and 0.7%, respectively). The pH and total titratable acid levels of rice wine according to the lactic acid concentration were stable during fermentation, according to the increase in the amount of lactic acid. The organic acids in the rice wines were highest in the lactic acid. The rice wines to which lactic acid had been added had lower acetic acid contents than the control. Also, the acetic acid contents decreased along with the lactic acid concentrations, except in the rice wine treated with 1.0% lactic acid. The ethanol contents of the tested rice wines were not significant. All in all, in the sensory evaluation, the rice wines treated with 0.5 and 0.7% lactic acid scored higher than the other treatments.

Combined effects of high pressure homogenization treatment and citral on microbiological quality of apricot juice

High pressure homogenization (HPH) technique is able to significantly reduce spoilage microbiota in fruit juice. On the other hand, aroma compounds and essential oils can have a key role in the microbial stability of these products. For this reason, the aim of this work was to evaluate the combined effects of an aroma compound (citral, used at a concentration of 50mg/l) and HPH treatments (performed at 100MPa for 1–8 successive passes) on the inactivation dynamics of Saccharomyces cerevisiae SPA strain inoculated in apricot juices at level of about 4.5 log CFU/ml. Moreover, growth of surviving yeast cells was measured during the storage of the treated juice at 10°C and pH, water activity, viscosity and volatile molecule profile of apricot juice were studied. Since citral had been diluted in ethanol before the addition to juice, also samples with only ethanol added at the same volume used to dissolve citral were considered. The results showed that yeast cell viability decreased with the increases of passes at 100MPa and the relationship between yeast cell loads and number of passes at 100MPa followed a linear trend.In addition, the effect of HPH treatment can be notably potentiated throughout the presence of citral and ethanol, increasing the time necessary to reach a spoilage threshold during storage. The volatile profiles of the juices added with citral showed a substitution by yeast metabolism of this aldehyde with molecule characterized by a lower antimicrobial activity such as alcohols. The HPH treatments had also a significant effect on pH and viscosity of apricot juices while did not affect aw.

Critical factors affecting the adherence of Candida albicans to the vaginal epithelium

A clinical isolate (Candida albicans) was collected from diagnosed cases as vaginal candidiasis. Identification of this isolate was previously reported [11] using both API 20C kit and real-time PCR assay. Adherence of C. albicans to the vaginal epithelium was evaluated. C. albicans recorded the highest rate (63.00%) of adherence among all collected isolates and recorded the highest record for the number (131.00) of adhered yeast cells (data not shown). The evaluation of the critical factors affecting the adherence process revealed that the maximum adherence values were recorded for a logarithmic phase-culture grown at 25°C, incubation temperature of 37°C in phosphate buffer saline adjusted at pH 5. Moreover, the adherence pattern of C. albicans cells to the vaginal epithelium was visualized by the scanning electron microscopy (SEM).

THE STUDY OF THE INFLUENCE OF TEMPERATURE AND INITIAL GLUCOSE CONCENTRATION ON THE FERMENTATION PROCESS IN THE PRESENCE OF Saccharomyces cerevisiae YEAST STRAIN IMMOBILIZED ON STARCH GELS BY REVERSED-FLOW GAS CHROMATOGRAPHY

The technique of reversed-flow gas chromatography (RFGC) was employed for the determination of the alcoholic fermentation phases and of kinetic parameters for free and immobilized cell systems, at different initial glucose concentrations and temperature values. In addition to this, due to its considerable advantages over other techniques, RFGC was used for the characterization of a new biocatalyst, yeast cells immobilized on starch gel, and especially wheat starch gel. Immobilization of wine yeast Saccharomyces cerevisiae AXAZ-1 was accomplished on wheat and corn starch gels in order to prepare new biocatalysts with great interest for the fermentation industry. The RFGC led with great accuracy, resulting from a literature review, to the determination of reaction rate constants and activation energies at each phase of the fermentation processes. A maximum value of rate constants was observed at initial glucose concentration of 205 g/L, where a higher number of yeast cells was observed. The increase of glucose concentrations had a negative influence on the growth of AXAZ-1 cells and rate constants were decreased. The decrease of fermentation temperature caused a substantial reduction in the viability of immobilized cells as well as in rate constant values. Activation energies of corn starch gel presented lower values than those of wheat starch gel. However, the two supports showed higher catalytic efficiency than free cell systems, proving that starch gels may act as a promoter of the catalytic activity of the yeast cells involved in the fermentation process.

Evaluation of the photocatalytic antimicrobial effects of a TiO2 nanocomposite food packaging film by in vitro and in vivo tests

The ability to inactivate Pseudomonas spp. and Rhodotorula mucilaginosa by a photocatalyst thin film, obtained via a combination of anatase and rutile titanium dioxide incorporation into a low density polyethylene (LDPE) polymeric matrix by a melt blending method, was addressed in this study. The number of surviving cell of the Pseudomonas spp. was decreased by 4 log CFU/ml and 1.35 log CFU/ml after 3 h of UVA illumination on TiO2 nanocomposite thin film and LDPE thin film, respectively. The number of cells of R. mucilaginosa decreased by 2 log CFU/ml and 0.64 log CFU/ml on TiO2 nanocomposite thin film and LDPE thin film, respectively. In an in vivo test carried out on fresh pears packaged in TiO2 nanocomposite film and stored under illumination by a fluorescent light lamp at 5 °C for 17 days, the number of mesophilic bacteria and yeast cells decreased significantly compared to samples stored in LDPE film. The greatest effects were recorded by combining UVA illumination and active film. It was also proven that the photocatalyst thin film prepared by extrusion could be effectively used in fruit packaging applications.

Capsule Independent Uptake of the Fungal Pathogen Cryptococcus neoformans into Brain Microvascular Endothelial Cells

Cryptococcosis is a life-threatening fungal disease with a high rate of mortality among HIV/AIDS patients across the world. The ability to penetrate the blood-brain barrier (BBB) is central to the pathogenesis of cryptococcosis, but the way in which this occurs remains unclear. Here we use both mouse and human brain derived endothelial cells (bEnd3 and hCMEC/D3) to accurately quantify fungal uptake and survival within brain endothelial cells. Our data indicate that the adherence and internalisation of cryptococci by brain microvascular endothelial cells is an infrequent event involving small numbers of cryptococcal yeast cells. Interestingly, this process requires neither active signalling from the fungus nor the presence of the fungal capsule. Thus entry into brain microvascular endothelial cells is most likely a passive event that occurs following ‘trapping’ within capillary beds of the BBB.

Trapping Yeast Cells on PDMS Micropillar Array

A selectively yeast cell trapping and arraying method was presented, utilizing PDMS micropillar array combining with chemical adhesion. Yeast cells were trapped on the top of micropillar array while swept on the flat area for the pinning of liquid/surface contact line on the micropillars and moving on the flat surface. And the modification of poly-L-lysine on the yeast cells improved the immobilization of cells on the surface of PDMS surface. Both of simulation and experiment results shows that by adjusting the diameter of micropillars, the number of yeast cells on each pillars could be controlled. Single cell array was formed with a 8.3 μm micropillar array, and majority yeast numbers of 3,4,5 was got for the 13.7 μm、18.0 μm and 18.8 μm micropillar array.

벌꿀을 이용한 고액분리 숙성 탁주의 주질 향상과 고형분의 조절에 따른 품질유지기간 증대

This study was conducted to improve quality of takju where natural honey is used to control the ripening fermentation and the amount of sediment derived from takju mash. A koji was prepared using rice starch and Aspergillus awamori var. kawachii. Takju mash was prepared by alcohol fermentation with Saccharomyces cerevisiae and a 3-step addition of steamed rice. The clean part of the mash (CPM) was separated from the sediment at and 5% (w/v) of natural honey was added and then ripened for 23 days at 5, 10, or . Temperature, pH, acidity, and total sugar content showed no significant differences, but a 0.2 percent reduction in alcohol content occurred during storage. However, CPM ripened with honey had a comparatively higher score on sensory evaluation than did immature CPM with added honey added. Takju with 8% alcohol content was prepared by mixing the water from the mixed CPM ripened with honey together with 100, 50, and 25% of the frozen sediment. Several quality characteristics of the takju were checked over 37 days of fermentation at . The pH was sustained between 4.1 and 4.3, and changes in the number of viable yeast cells, acidity, total sugar amounts, and alcohol content showed similar patterns but differences in scale. Smaller amounts of sediment affected the stability of the takju. Mixing the CPM ripened with natural honey at low temperature moderately reduced the amount of sediment in the mash and resulted in a highly flavorful takju with an extended shelf life.

Ethanol production potential from fermented rice noodle wastewater treatment using entrapped yeast cell sequencing batch reactor

Fermented rice noodle production generates a large volume of starch-based wastewater. This study investigated the treatment of the fermented rice noodle wastewater using entrapped cell sequencing batch reactor (ECSBR) compared to traditional sequencing batch reactor (SBR). The yeast cells were applied because of their potential to convert reducing sugar in the wastewater to ethanol. In present study, preliminary treatment by acid hydrolysis was performed. A yeast culture, Saccharomyces cerevisiae, with calcium alginate cell entrapment was used. Optimum yeast cell loading in batch experiment and fermented rice noodle treatment performances using ECSBR and SBR systems were examined. In the first part, it was found that the cell loadings (0.6–2.7 × 108 cells/mL) did not play an important role in this study. Treatment reactions followed the second-order kinetics with the treatment efficiencies of 92–95%. In the second part, the result showed that ECSBR performed better than SBR in both treatment efficiency and system stability perspectives. ECSBR maintained glucose removal of 82.5 ± 10% for 5-cycle treatment while glucose removal by SBR declined from 96 to 40% within the 5-cycle treatment. Scanning electron microscopic images supported the treatment results. A number of yeast cells entrapped and attached onto the matrix grew in the entrapment matrix.

Artificial neural network approach to modeling of alcoholic fermentation of thick juice from sugar beet processing

In this paper the bioethanol production in batch culture by free Saccharomyces cerevisiae cells from thick juice as intermediate product of sugar beet processing was examined. The obtained results suggest that it is possible to decrease fermentation time for the cultivation medium based on thick juice with starting sugar content of 5-15 g kg-1. For the fermentation of cultivation medium based on thick juice with starting sugar content of 20 and 25 g kg-1 significant increase in ethanol content was attained during the whole fermentation process, resulting in 12.51 and 10.95 dm3 m-3 ethanol contents after 48 h, respectively. Other goals of this work were to investigate the possibilities for experimental results prediction using artificial neural networks (ANNs) and to find its optimal topology. A feed-forward back-propagation artificial neural network was used to test the hypothesis. As input variables fermentation time and starting sugar content were used. Neural networks had one output value, ethanol content, yeast cell number or sugar content. There was one hidden layer and the optimal number of neurons was found to be nine for all selected network outputs. In this study transfer function was tansig and the selected learning rule was Levenberg-Marquardt. Results suggest that artificial neural networks are good prediction tool for selected network outputs. It was found that experimental results are in very good agreement with computed ones. The coefficient of determination (the R-squared) was found to be 0.9997, 0.9997 and 0.9999 for ethanol content, yeast cell number and sugar content, respectively.

Phosphatidate Phosphatase Plays Role in Zinc-mediated Regulation of Phospholipid Synthesis in Yeast

In the yeast Saccharomyces cerevisiae, the synthesis of phospholipids is coordinately regulated by mechanisms that control the homeostasis of the essential mineral zinc (Carman, G.M., and Han, G. S. (2007) Regulation of phospholipid synthesis in Saccharomyces cerevisiae by zinc depletion. Biochim. Biophys. Acta 1771, 322-330; Eide, D. J. (2009) Homeostatic and adaptive responses to zinc deficiency in Saccharomyces cerevisiae. J. Biol. Chem. 284, 18565-18569). The synthesis of phosphatidylcholine is balanced by the repression of CDP-diacylglycerol pathway enzymes and the induction of Kennedy pathway enzymes. PAH1-encoded phosphatidate phosphatase catalyzes the penultimate step in triacylglycerol synthesis, and the diacylglycerol generated in the reaction may also be used for phosphatidylcholine synthesis via the Kennedy pathway. In this work, we showed that the expression of PAH1-encoded phosphatidate phosphatase was induced by zinc deficiency through a mechanism that involved interaction of the Zap1p zinc-responsive transcription factor with putative upstream activating sequence zinc-responsive elements in the PAH1 promoter. The pah1Δ mutation resulted in the derepression of the CHO1-encoded phosphatidylserine synthase (CDP-diacylglycerol pathway enzyme) and loss of the zinc-mediated regulation of the enzyme. Loss of phosphatidate phosphatase also resulted in the derepression of the CKI1-encoded choline kinase (Kennedy pathway enzyme) but decreased the synthesis of phosphatidylcholine when cells were deficient of zinc. This result confirmed the role phosphatidate phosphatase plays in phosphatidylcholine synthesis via the Kennedy pathway.

The role of adjuvants in therapeutic protection against paracoccidioidomycosis after immunization with the P10 peptide

Paracoccidioidomycosis (PCM), a common chronic mycosis in Latin America, is a granulomatous systemic disease caused by the thermo-dimorphic fungus Paracoccidioides brasiliensis. The glycoprotein gp43 is the main antigen target of P. brasiliensis and a 15-mer internal peptide (QTLIAIHTLAIRYAN), known as P10, defines a major CD4+-specific T cell epitope. Previous results have indicated that, besides having a preventive role in conventional immunizations prior to challenge with the fungus, protective anti-fungal effects can be induced in P. brasiliensis-infected mice treated with P10 administered with complete Freund’s adjuvant (CFA). The peptide elicits an IFN-γ-dependent Th1 immune response and is the main candidate for effective immunotherapy of patients with PCM, as an adjunctive approach to conventional chemotherapy. In the present study we tested the therapeutic effects of P10 combined with different adjuvants [aluminum hydroxide, CFA, flagellin, and the cationic lipid dioctadecyl-dimethylammonium bromide (DODAB)] in BALB/c mice previously infected with the P. brasiliensis Pb18 strain. Significant reductions in the number of colony forming units of the fungus were detected in lungs of mice immunized with P10 associated with the different adjuvants 52 days after infection. Mice treated with DODAB and P10, followed by mice treated with P10 and flagellin, showed the most prominent effects as demonstrated by the lowest numbers of viable yeast cells as well as reductions in granuloma formation and fibrosis. Concomitantly, secretion of IFN-γ and TNF-α, in contrast to interleukin (IL)-4 and IL-10, was enhanced in the lungs of mice immunized with P10 in combination with the tested adjuvants, with the best results observed in mice treated with P10 and DODAB. In conclusion, the present results demonstrate that the co-administration of the synthetic P10 peptide with several adjuvants, particularly DODAB, have significant therapeutic effects in experimental PCM.

Autoantibody detection by direct counting of antigen-displayed yeast cells

In this study, we report a new immunoassay platform based on yeast surface display technology for detection of autoantibodies involved in autoimmune diseases, e.g., systemic lupus erythematosus (SLE) and Sjögren's syndrome (SS). The autoantigens of Ro52/SSA epitope and SmD were chosen to be displayed on the yeast surface with their respective antibodies as the analytes. By using magnetic beads modified with protein G, yeast cells bound with specific target antibody can be captured. The amount of analytes could be determined by counting the number of fluorescent yeast cells captured in a magnetic field. The platform showed promising results in the detection of SLE autoantibodies with high sensitivity and multiplex detection capability over the traditional approaches.

Yeast DNA Replication 2D Gel Protocol

Two-dimensional agarose gel electrophoresis (2D gel) analysis is used extensively as a method to detect origins of replication. Here, I present a simplified method for the isolation of yeast genomic DNA for 2D gel analysis from a small number of yeast cells. This DNA isolation method is simpler and less time consuming than the traditional method that involves CsCl density gradient centrifugation. This method could be modified for 2D gel analysis in other organisms as well.

Concanavalin-A induces IL-17 production during the course of Candida albicans infection

In a previous study, our group verified that 100% of mice survived to a lethal dose of Candida albicans following pretreatment with concanavalin-A (Con-A) for 3 days. This work proposed to investigate whether treatment could mediate an adaptative immune response involving T(H) 17 cells. A significant increase in IL-17 levels at 6 h postinfection was observed and was maintained up to 18 h in the Con-A group, whereas in control mice, a reduction in this cytokine was verified. In addition, T(H) 17 cells develop in the presence of TGF-β, IL-1 β, and IL-6 that were increased significantly 2 h postinfection in Con-A-treated mice. Macrophages were involved in the process, engulfing greater numbers of yeast cells, and were activated through TNF-α and interferon-γ produced at significant levels at 2 h postinfection. A significant increase in IL-12 levels was also observed at 2 h postinfection. Thus, activated macrophages were probably more capable of killing and processing Candida antigens, signalizing an adaptative immune response. Macrophages from controls did not prevent yeast-to-hyphae transition and were partially destroyed, as shown in scanning microscopy. These results suggest that treatment with Con-A facilitated the triggering of T(H) 17 and T(H) 1 responses via IL-17 and IFN-γ production, leading to the resolution of C. albicans infection.