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International Journal of Occupational... Jun 2013Yeasts may become potential human and animal pathogens, particularly for individuals with a depressed immune system. Their presence in the environment, especially in...
OBJECTIVES
Yeasts may become potential human and animal pathogens, particularly for individuals with a depressed immune system. Their presence in the environment, especially in soil, may favour their spread into human ontocenoses.
MATERIALS AND METHODS
Eighty-four soil samples obtained from 21 children's recreational sites in Łódź in autumn 2010 and spring 2011 were evaluated. The yeasts were isolated by classical microbiological methods and identified on the basis of morphological and biochemical features.
RESULTS
The fungi were found in 73.8% and in 69.0% of the examined samples collected in autumn and spring, respectively. Among 97 isolates of yeasts, the species potentially pathogenic to humans and animals were Candida colliculosa, C. guilliermondii, C. humicola, C. inconspicua, C. lambica, C. lusitaniae, C. pelliculosa, C. tropicalis, Cryptococcus albidus, C. laurentii, C. neoformans, C. terreus, Kloeckera japonica, Geotrichum candidum, G. penicillatum, Rhodotorula mucilaginosa, R. glutinis, Saccharomyces cerevisiae, Sporobolomyces salmonicolor and Trichosporon cutaneum. The most frequently isolated fungi included the genus Cryptococcus (38 isolates) and two species: Rhodotorula glutinis (15), Trichosporon cutaneum (14). C. neoformans, an etiological factor of cryptococcal meningitis, was present in the sandpits of 3 kindergartens. The Candida species were identified from park playgrounds and school sports fields mainly in autumn 2010 (14 isolates), in spring 2011 - only 1 isolate. The concentration of fungal species in particular samples varied considerably, but in the majority of samples, fungi were present at concentration of up to 1×10(2) CFU/1 g of soil.
CONCLUSIONS
Yeasts were present in the soil of parks, schools and kindergarten recreational areas; the fact may pose a health risk to humans, especially to children, and this type of biological pollution should be regarded as a potential public health concern.
Topics: Candida; Child; Cities; Cryptococcus; Environment; Geotrichum; Humans; Kloeckera; Poland; Recreation; Rhodotorula; Saccharomyces; Schools; Seasons; Trichosporon; Yeasts
PubMed: 24018998
DOI: 10.2478/s13382-013-0118-y -
Microbial Cell Factories Jun 2013Oats contain hydroxycinnamoyl anthranilates, also named avenanthramides (Avn), which have beneficial health properties because of their antioxidant, anti-inflammatory,...
BACKGROUND
Oats contain hydroxycinnamoyl anthranilates, also named avenanthramides (Avn), which have beneficial health properties because of their antioxidant, anti-inflammatory, and antiproliferative effects. The microbial production of hydroxycinnamoyl anthranilates is an eco-friendly alternative to chemical synthesis or purification from plant sources. We recently demonstrated in yeast (Saccharomyces cerevisiae) that coexpression of 4-coumarate: CoA ligase (4CL) from Arabidopsis thaliana and hydroxycinnamoyl/benzoyl-CoA/anthranilate N-hydroxycinnamoyl/benzoyltransferase (HCBT) from Dianthus caryophyllusenabled the biological production of several cinnamoyl anthranilates upon feeding with anthranilate and various cinnamates. Using engineering strategies to overproduce anthranilate and hydroxycinnamates, we describe here an entire pathway for the microbial synthesis of two Avns from glucose in Escherichia coli.
RESULTS
We first showed that coexpression of HCBT and Nt4CL1 from tobacco in the E. coli anthranilate-accumulating strain W3110 trpD9923 allowed the production of Avn D [N-(4'-hydroxycinnamoyl)-anthranilic acid] and Avn F [N-(3',4'-dihydroxycinnamoyl)-anthranilic acid] upon feeding with p-coumarate and caffeate, respectively. Moreover, additional expression in this strain of a tyrosine ammonia-lyase from Rhodotorula glutinis (RgTAL) led to the conversion of endogenous tyrosine into p-coumarate and resulted in the production of Avn D from glucose. Second, a 135-fold improvement in Avn D titer was achieved by boosting tyrosine production using two plasmids that express the eleven genes necessary for tyrosine synthesis from erythrose 4-phosphate and phosphoenolpyruvate. Finally, expression of either the p-coumarate 3-hydroxylase Sam5 from Saccharothrix espanensis or the hydroxylase complex HpaBC from E. coli resulted in the endogenous production of caffeate and biosynthesis of Avn F.
CONCLUSION
We established a biosynthetic pathway for the microbial production of valuable hydroxycinnamoyl anthranilates from an inexpensive carbon source. The proposed pathway will serve as a platform for further engineering toward economical and sustainable bioproduction of these pharmaceuticals and other related aromatic compounds.
Topics: Acyltransferases; Ammonia-Lyases; Arabidopsis; Biosynthetic Pathways; Caffeic Acids; Coenzyme A Ligases; Dianthus; Escherichia coli; Fungal Proteins; Genetic Engineering; Glucose; Mixed Function Oxygenases; Plant Proteins; Plasmids; Rhodotorula; Tyrosine; ortho-Aminobenzoates
PubMed: 23806124
DOI: 10.1186/1475-2859-12-62 -
3 Biotech Feb 2013Rhodotorula glutinis CCY 20-2-26 when grown under controlled stress of either NaCl (1-5 %) or HO (1-5 mM) on basal media exhibited a twofold increase in its total...
Rhodotorula glutinis CCY 20-2-26 when grown under controlled stress of either NaCl (1-5 %) or HO (1-5 mM) on basal media exhibited a twofold increase in its total phenolic contents. The radical scavenging capacities (RSCs) as determined by ABTS test were found to be highest in 4 mM HO (1.44 mM TEAC mg) and 4 % NaCl (1.13 mM TEAC mg) as compared to control samples (0.41 mM TEAC mg). Similarly, the RSCs as determined by DPPH test were also highest in 4 % NaCl (1.83 mM TEAC mg) and 4 mM HO (1.78 mM TEAC mg) compared to control (0.48 TEAC mg). The relative RSCs from EPR spin-trapping assay for HO-stressed cultures were highest in 1 mM HO (56.1 μM TEAC g) whereas in NaCl-stressed cultures it was highest in 5 % NaCl (44.6 μM TEAC g) as compared to control (30.9 μM TEAC g). Five phenolic compounds (gallic acid, benzoic acid, catechin, caffeic acid and ferulic acid) were detected for the first time in R. glutinis CCY 20-2-26.
PubMed: 28324345
DOI: 10.1007/s13205-012-0069-1 -
Microbiology (Reading, England) Jan 2013The biosynthesis of triacylglycerol (TAG) occurs in the microsomal membranes of eukaryotes. Here, we report the identification and functional characterization of...
The biosynthesis of triacylglycerol (TAG) occurs in the microsomal membranes of eukaryotes. Here, we report the identification and functional characterization of diacylglycerol acyltransferase (DGAT), a member of the 10 S cytosolic TAG biosynthetic complex (TBC) in Rhodotorula glutinis. Both a full-length and an N-terminally truncated cDNA clone of a single gene were isolated from R. glutinis. The DGAT activity of the protein encoded by RgDGAT was confirmed in vivo by the heterologous expression of cDNA in a Saccharomyces cerevisiae quadruple mutant (H1246) that is defective in TAG synthesis. RgDGAT overexpression in yeast was found to be capable of acylating diacylglycerol (DAG) in an acyl-CoA-dependent manner. Quadruple mutant yeast cells exhibit growth defects in the presence of oleic acid, but wild-type yeast cells do not. In an in vivo fatty acid supplementation experiment, RgDGAT expression rescued quadruple mutant growth in an oleate-containing medium. We describe a soluble acyl-CoA-dependent DAG acyltransferase from R. glutinis that belongs to the DGAT3 class of enzymes. The study highlights the importance of an alternative TAG biosynthetic pathway in oleaginous yeasts.
Topics: Acyl Coenzyme A; Acylation; Cloning, Molecular; Cluster Analysis; DNA, Fungal; Diacylglycerol O-Acyltransferase; Diglycerides; Gene Expression; Molecular Sequence Data; Phylogeny; Rhodotorula; Saccharomyces cerevisiae; Sequence Analysis, DNA; Triglycerides
PubMed: 23103975
DOI: 10.1099/mic.0.063156-0 -
Journal of Clinical Microbiology Sep 2011Candida auris is a newly described species whose clinical significance is not clear. Here, we describe the first three cases of nosocomial fungemia caused by C. auris,...
Candida auris is a newly described species whose clinical significance is not clear. Here, we describe the first three cases of nosocomial fungemia caused by C. auris, which confirms that it is a causative agent of bloodstream infections. All three patients presented persistent fungemia for 10 to 31 days. The isolates obtained from the three patients were misidentified as Candida haemulonii and Rhodotorula glutinis by the Vitek 2 and the API 20C systems, respectively. C. auris was confirmed by sequence analysis of the internal transcribed spacer region and D1/D2 regions of the 26S ribosomal DNA of the rRNA gene. The MIC ranges of amphotericin B (AMB), fluconazole (FLU), itraconazole, and voriconazole were 0.5 to 1, 2 to 128, 0.125 to 2, and 0.06 to 1 μg/ml, respectively. All isolates were susceptible to caspofungin (MIC = 0.06 μg/ml) and micafungin (MIC = 0.03 μg/ml). One patient developed breakthrough fungemia while receiving FLU therapy, and two patients who received FLU therapy followed by AMB showed therapeutic failure and fatal outcomes. Our cases show that C. auris fungemia can be persistent, despite FLU or AMB therapy, which emphasizes the importance of accurately identifying this species.
Topics: Aged; Antifungal Agents; Candida; Candidemia; Cross Infection; DNA, Fungal; DNA, Ribosomal Spacer; Diagnostic Errors; Female; Humans; Infant; Male; Microbial Sensitivity Tests; Mycological Typing Techniques; Sequence Analysis, DNA
PubMed: 21715586
DOI: 10.1128/JCM.00319-11 -
Enzyme Research 2011Enantioselective reductions of p-R(1)-C(6)H(4)C(O)CH(2)R(2) (R(1) = Cl, Br, CH(3), OCH(3), NO(2) and R(2) = Br, Cl) mediated by Geotrichum candidum CCT 1205 and...
Chiral Pharmaceutical Intermediaries Obtained by Reduction of 2-Halo-1-(4-substituted phenyl)-ethanones Mediated by Geotrichum candidum CCT 1205 and Rhodotorula glutinis CCT 2182.
Enantioselective reductions of p-R(1)-C(6)H(4)C(O)CH(2)R(2) (R(1) = Cl, Br, CH(3), OCH(3), NO(2) and R(2) = Br, Cl) mediated by Geotrichum candidum CCT 1205 and Rhodotorula glutinis CCT 2182 afforded the corresponding halohydrins with complementary R and S configurations, respectively, in excellent yield and enantiomeric excesses. The obtained (R)- or (S)-halohydrins are important building blocks in chemical and pharmaceutical industries.
PubMed: 21687613
DOI: 10.4061/2011/976368 -
Journal of Applied Microbiology Apr 2011We developed improved methods for DNA-based fluorescence in situ hybridization (FISH) for rapid detection of Candida spp. and Candida albicans via flow cytometry.
AIMS
We developed improved methods for DNA-based fluorescence in situ hybridization (FISH) for rapid detection of Candida spp. and Candida albicans via flow cytometry.
METHODS AND RESULTS
Two previously reported C. albicans-targeted DNA probes were evaluated against whole cells of C. albicans and related Candida species using a rapid, high-temperature hybridization protocol. One probe (CalB2208) was shown for the first time to be suitable as a FISH probe. Although cell labelling for both probes was relatively bright, we were able to substantially improve our results by altering fixation and hybridization conditions. For fixation, a 60 : 40 mixture of 10% buffered formalin and ethanol was most effective. Probe intensity was improved as much as ten-fold through the use of unlabelled helper probes, and buffer containing 0·9 mol l−1 NaCl plus 10% formamide yielded the best hybridizations for both probe/helper cocktails. Although optimal labelling occurred with longer hybridizations, we found that C. albicans could be completely differentiated from the nontarget yeast Rhodotorula glutinis after only 15 min using the brightest probe (Calb-1249) and that a formal washing step was not required. Specificities of probe/helper cocktails under optimal conditions were determined using a panel of target and nontarget cell types, including four strains of Candida dubliniensis. Calb-1249 cross-reacted slightly with Candida parapsilosis and strongly with both Candida tropicalis and C. dubliniensis. In contrast, we found that CalB2208 was exclusive for C. albicans. The molecular basis of this specificity was confirmed by DNA sequencing.
CONCLUSIONS
We describe DNA probe-based approaches for rapid and bright labelling of Candida spp. and for specific labelling of C. albicans without cross-reaction with C. dubliniensis. Our work improves upon previously described methods.
Topics: Candida; Candida albicans; Candida tropicalis; DNA Probes; Flow Cytometry; In Situ Hybridization, Fluorescence; Nucleic Acid Hybridization; Sequence Analysis, DNA
PubMed: 21205104
DOI: 10.1111/j.1365-2672.2011.04936.x -
Le Infezioni in Medicina Jun 2010Fungal sepsis is an important cause of fever resistant to antibiotic therapy that is very often taken into marginal account. It should instead be particularly considered...
Fungal sepsis is an important cause of fever resistant to antibiotic therapy that is very often taken into marginal account. It should instead be particularly considered in patients with a long history of immune depression such as diabetes or chronic and debilitating diseases. Blood cultures are essential for diagnostic purposes, preferably performed in antibiotic wash-out, since they may allow identification of pathogenic (or opportunistic) fungi responsible for episodes of fungal sepsis. The case described illustrates an episode of systemic infection by Rhodotorula glutinis correlated with the presence of CVC.
Topics: Aged; Antifungal Agents; Catheter-Related Infections; Catheterization, Central Venous; Communicable Diseases, Emerging; Comorbidity; Diabetes Complications; Fungemia; Humans; Immunocompromised Host; Liver Cirrhosis; Male; Mycoses; Pyrimidines; Rhodotorula; Sepsis; Triazoles; Voriconazole
PubMed: 20610937
DOI: No ID Found -
Annals of Clinical Microbiology and... Mar 2010The aim of this study was to investigate the presence of multidrug resistant yeasts in the faeces of synanthropic wild birds from the Bangsar suburb of Kuala Lumpur.
BACKGROUND
The aim of this study was to investigate the presence of multidrug resistant yeasts in the faeces of synanthropic wild birds from the Bangsar suburb of Kuala Lumpur.
METHODS
Species characterisations of yeast isolates and determinations of antimycotic susceptibility profiles were undertaken using the commercial characterization kit, Integral System Yeasts Plus (Liofilchem, Italy).
RESULTS
Fourteen species of yeasts were detected in the bird faecal samples.Candida albicans was present in 28.89% of bird faecal samples, Candida krusei (13.33%), Candida tropicalis (4.44%), Candida glabrata (4.44%), Candida parapsilosis (2.22%), Candida lambica (2.22%), Candida stellatoidea (2.22%), Candida rugosa (2.22%) and Candida lusitaniae (2.22%). Amongst the non-candidal yeast isolates, Cryptococcus laurentii was present in 6.67% of bird faecal samples, Cryptococcus uniguttulatus (4.44%), Saccharomyces cerevisiae (4.44%), Trichosporon pullulans (2.22%), Trichosporon pullulans/Cryptococcus albidus (8.89%) and Rhodotorula rubra/Rhodotorula glutinis (4.44%). Of the isolated yeasts, 18.1% (or 26/144) were found to be resistant to all 11 antimycotic agents they were tested against i.e. Nystatin, Amphotericin B, Flucytosine, Econazole, Ketoconazole, Clotrimazole, Miconazole, Itraconazole, Voriconazole, Fluconazole 16 and Fluconazole 64. 45.8% (or 66/144) of the bird faecal yeast isolates were resistant to four or more of the 11 antimycotic agents they were tested against.
CONCLUSIONS
This finding is of public health significance as these synanthropic wild birds may be reservoirs for transmission of drug resistant yeast infections to humans.
Topics: Animals; Antifungal Agents; Birds; Drug Resistance, Multiple, Fungal; Environmental Monitoring; Face; Humans; Malaysia; Mycoses; Yeasts
PubMed: 20307325
DOI: 10.1186/1476-0711-9-11 -
Journal of Applied Microbiology May 2010To isolate and characterize microbes in the soils containing high contents of phenolics and to dissolve the allelopathic inhibition of plants through microbial...
AIMS
To isolate and characterize microbes in the soils containing high contents of phenolics and to dissolve the allelopathic inhibition of plants through microbial degradation.
METHODS AND RESULTS
Four microbes were isolated from plant soils using a screening medium containing p-coumaric acid as sole carbon source. The isolates were identified by biochemical analysis and sequences of their 16S or 18S rDNA, and designated as Pseudomonas putida 4CD1 from rice (Oryza sativa) soil, Ps. putida 4CD3 from pine (Pinus massoniana) soil, Pseudomonas nitroreducens 4CD2 and Rhodotorula glutinis 4CD4 from bamboo (Bambusa chungii) soil. All isolates degraded 1 g l(-1) of p-coumaric acid by 70-93% in inorganic and by 99% in Luria-Bertani solutions within 48 h. They also effectively degraded ferulic acid, p-hydroxybenzoic acid and p-hydroxybenzaldehyde. The microbes can degrade p-coumaric acid and reverse its inhibition on seed germination and seedling growth in culture solutions and soils. Low pHs inhibited the growth and phenolic degradation of the three bacteria. High temperature inhibited the R. glutinis. Co(2+) completely inhibited the three bacteria, but not the R. glutinis. Cu(2+), Al(3+), Zn(2+), Fe(3+), Mn(2+), Mg(2+) and Ca(2+) had varying degrees of inhibition for each of the bacteria.
CONCLUSIONS
Phenolics in plant culture solutions and soils can be decomposed through application of soil microbes in laboratory or controlled conditions. However, modification of growth conditions is more important for acidic and ions-contaminated media.
SIGNIFICANCE AND IMPACT OF THE STUDY
The four microbes were first isolated and characterized from the soils of bamboo, rice or pine. This study provides some evidence and methods for microbial control of phenolic allelochemicals.
Topics: Coumaric Acids; Germination; Hydrogen-Ion Concentration; Metals; Oryza; Phenols; Pheromones; Pseudomonas; RNA, Ribosomal, 16S; RNA, Ribosomal, 18S; Rhodotorula; Sasa; Soil Microbiology; Temperature
PubMed: 19912433
DOI: 10.1111/j.1365-2672.2009.04589.x