Proteomic and Metabolomic Analyses Reveal Contrasting Anti-Inflammatory Effects of an Extract of Mucor Racemosus Secondary Metabolites Compared to Dexamethasone.

PubMed

Meier, Samuel M; Muqaku, Besnik; Ullmann, Ronald; Bileck, Andrea; Kreutz, Dominique; Mader, Johanna C; Knasmüller, Siegfried; Gerner, Christopher

2015-01-01

Classical drug assays are often confined to single molecules and targeting single pathways. However, it is also desirable to investigate the effects of complex mixtures on complex systems such as living cells including the natural multitude of signalling pathways. Evidence based on herbal medicine has motivated us to investigate potential beneficial health effects of Mucor racemosus (M rac) extracts. Secondary metabolites of M rac were collected using a good-manufacturing process (GMP) approved production line and a validated manufacturing process, in order to obtain a stable product termed SyCircue (National Drug Code USA: 10424-102). Toxicological studies confirmed that this product does not contain mycotoxins and is non-genotoxic. Potential effects on inflammatory processes were investigated by treating stimulated cells with M rac extracts and the effects were compared to the standard anti-inflammatory drug dexamethasone on the levels of the proteome and metabolome. Using 2D-PAGE, slight anti-inflammatory effects were observed in primary white blood mononuclear cells, which were more pronounced in primary human umbilical vein endothelial cells (HUVECs). Proteome profiling based on nLC-MS/MS analysis of tryptic digests revealed inhibitory effects of M rac extracts on pro-inflammatory cytoplasmic mediators and secreted cytokines and chemokines in these endothelial cells. This finding was confirmed using targeted proteomics, here treatment of stimulated cells with M rac extracts down-regulated the secretion of IL-6, IL-8, CXCL5 and GROA significantly. Finally, the modulating effects of M rac on HUVECs were also confirmed on the level of the metabolome. Several metabolites displayed significant concentration changes upon treatment of inflammatory activated HUVECs with the M rac extract, including spermine and lysophosphatidylcholine acyl C18:0 and sphingomyelin C26:1, while the bulk of measured metabolites remained unaffected. Interestingly, the effects of M rac

Rhinocerebral Mucor circinelloides infection in immunocompromised patient following yogurt ingestion.

PubMed

Lazar, Stephen P; Lukaszewicz, Jennifer M; Persad, Kamleish A; Reinhardt, John F

2014-08-01

The purpose of this case report is to illustrate the cause of this patient's headache and sinus pain in the setting of a unique environmental exposure: the patient ingested yogurt only days before presentation. This particular brand of yogurt caused controversy in early September 2013 when the manufacturer voluntarily recalled all flavors. The yogurt was found to be contaminated with Mucor circinelloides. The recall was triggered by the FDA, after receiving many complaints from consumers affected by temporary gastrointestinal symptoms such as abdominal cramping, diarrhea, and nausea. This patient was diagnosed with Rhinocerebral mucormycosis through fungal culture of the affected area. He was specifically colonized with Mucor circinelloides, a variant that rarely causes disease in humans. According to a literature review, only eight cases of mucormycosis in adults caused by this strain were documented before 2009.

Microbiological Transformation of Enrofloxacin by the Fungus Mucor ramannianus

PubMed Central

Parshikov, Igor A.; Freeman, James P.; Lay, Jackson O.; Beger, Richard D.; Williams, Anna J.; Sutherland, John B.

2000-01-01

Enrofloxacin metabolism by Mucor ramannianus was investigated as a model for the biotransformation of veterinary fluoroquinolones. Cultures grown in sucrose-peptone broth were dosed with enrofloxacin. After 21 days, 22% of the enrofloxacin remained. Three metabolites were identified: enrofloxacin N-oxide (62% of the total absorbance), N-acetylciprofloxacin (8.0%), and desethylene-enrofloxacin (3.5%). PMID:10831454

Production of Mozzarella Cheese Using Rennin Enzyme from Mucor miehei Grown at Rice Bran Molasses Medium

NASA Astrophysics Data System (ADS)

Rusdan, I. H.; Kusnadi, J.

2017-04-01

The research aimed to study the characteristic and yield of Mozzarella cheese produced by using rennin enzyme from Mucor miehei which is grown at rice bran and molasses medium. The popularity of Mozzarella cheese in Indonesia is increased caused by the spreading of western foods in Indonesia such as pizza and spaghetti that use Mozzarella cheese for ingredient. In Italy, Mozzarella and pizza cheeses are dominating 78% of the total Italian Cheese products. In producing Mozzarella cheese, rennin enzyme is always used as milk coagulant. Even now, Indonesia has not produced the rennin enzyme yet. The rennin enzyme from Mucor miehei growing at rice bran and molases medium which have the availability can be managed purposively within short period of time. The completly randomized design methode used to get the best crude extracts of Mucor miehei rennin enzyme, then is employed to produce mozzarella cheese. The result of Mozzarella cheese has various characteristics such as the yield’s weight is 9.1%, which consists of 50% moisture content, 36.64% peotein levels, 0.1 melting ability and 82.72% stretch ability or 0.79/N. With that characteristic it is concluded that rennin enzyme from Mucor miehei grown at rice bran molasses medium has the potential to alternatively subtitute calf rennin to produce Mozzarella cheese, and the characteristics fulfill the standart.

Canthaxanthin production with modified Mucor circinelloides strains.

PubMed

Papp, Tamás; Csernetics, Arpád; Nagy, Gábor; Bencsik, Ottó; Iturriaga, Enrique A; Eslava, Arturo P; Vágvölgyi, Csaba

2013-06-01

Canthaxanthin is a natural diketo derivative of β-carotene primarily used by the food and feed industries. Mucor circinelloides is a β-carotene-accumulating zygomycete fungus and one of the model organisms to study the carotenoid biosynthesis in fungi. In this study, the β-carotene ketolase gene (crtW) of the marine bacterium Paracoccus sp. N81106 fused with fungal promoter and terminator regions was integrated into the M. circinelloides genome to construct stable canthaxanthin-producing strains. Different transformation methods including polyethylene glycol-mediated transformation with linear DNA fragments, restriction enzyme-mediated integration and Agrobacterium tumefaciens-mediated transformation were tested to integrate the crtW gene into the Mucor genome. Mitotic stability, site of integration and copy number of the transferred genes were analysed in the transformants, and several stable strains containing the crtW gene in high copy number were isolated. Carotenoid composition of selected transformants and effect of culturing conditions, such as temperature, carbon sources and application of certain additives in the culturing media, on their carotenoid content were analysed. Canthaxanthin-producing transformants were able to survive at higher growth temperature than the untransformed strain, maybe due to the effect of canthaxanthin on the membrane fluidity and integrity. With the application of glucose, trehalose, dihydroxyacetone and L-aspartic acid as sole carbon sources in minimal medium, the crtW-expressing M. circinelloides strain, MS12+pCA8lf/1, produced more than 200 μg/g (dry mass) of canthaxanthin.

Calcineurin orchestrates dimorphic transitions, antifungal drug responses and host-pathogen interactions of the pathogenic mucoralean fungus Mucor circinelloides.

PubMed

Lee, Soo Chan; Li, Alicia; Calo, Silvia; Inoue, Makoto; Tonthat, Nam K; Bain, Judith M; Louw, Johanna; Shinohara, Mari L; Erwig, Lars P; Schumacher, Maria A; Ko, Dennis C; Heitman, Joseph

2015-09-01

Calcineurin plays essential roles in virulence and growth of pathogenic fungi and is a target of the natural products FK506 and Cyclosporine A. In the pathogenic mucoralean fungus Mucor circinelloides, calcineurin mutation or inhibition confers a yeast-locked phenotype indicating that calcineurin governs the dimorphic transition. Genetic analysis in this study reveals that two calcineurin A catalytic subunits (out of three) are functionally diverged. Homology modeling illustrates modes of resistance resulting from amino substitutions in the interface between each calcineurin subunit and the inhibitory drugs. In addition, we show how the dimorphic transition orchestrated by calcineurin programs different outcomes during host-pathogen interactions. For example, when macrophages phagocytose Mucor yeast, subsequent phagosomal maturation occurs, indicating host cells respond appropriately to control the pathogen. On the other hand, upon phagocytosis of spores, macrophages fail to form mature phagosomes. Cytokine production from immune cells differs following exposure to yeast versus spores (which germinate into hyphae). Thus, the morphogenic transition can be targeted as an efficient treatment option against Mucor infection. In addition, genetic analysis (including gene disruption and mutational studies) further strengthens the understanding of calcineurin and provides a foundation to develop antifungal agents targeting calcineurin to deploy against Mucor and other pathogenic fungi. © 2015 John Wiley & Sons Ltd.

Chronic rhino-orbital mucormycosis caused by Mucor irregularis (Rhizomucor variabilis) in India

USDA-ARS?s Scientific Manuscript database

We describe a chronic case of rhino-orbital zygomycosis caused by Mucor irregularis, formerly known as Rhizomucor variabilis var. variabilis, a rare mycotic agent in humans. The infection caused progressive destruction of the nasal septum, soft and hard palate, leading to collapse of the nose bridge...

[Characterization of a malic enzyme isoform V from Mucor circinelloides].

PubMed

Zhang, Yingtong; Chen, Haiqin; Song, Yuanda; Zhang, Hao; Chen, Yongquan; Chen, Wei

2016-02-04

We aimed at characterizing a malic enzyme isoform V from Mucor circinelloides. me1 gene encoding malic enzyme isoform V was amplified and cloned into expression vector pET28a. High-purity recombinant protein BLME1 was obtained by affinity chromatography using. Ni-NTA column and characterized subsequently. The optimum conditions were pH at 8.0 and temperature at 33 degrees C. Under optimum conditions, BLME1 activity achieved 92.8 U/mg. The K(m) for L-malate and NADP+ were 0.74960 ± 0.06120 mmol/L and 0.22070 ± 0.01810 mmol/L, the V(max) for L-malate and NADP+ were 72.820 ± 1.077 U/mg and 86.110 ± 1.665 U/mg, respectively. In addition, ions played important roles in BLME1 activity; several ions such as Mn2+, Mg2+, Co2+, Ni2+ could activate BLME1, whereas Ca2+, Cu2+ could be used as inhibitors. Additionally, the metabolic intermediates such as oxaloacetic acid and α-ketoglutaric acid inhibited the activity of BLME1, whereas succinic acid activated it. A malic enzyme isoform V from Mucor circinelloides was characterized, providing the references for further studies on this enzyme.

Mucormycosis Caused by Unusual Mucormycetes, Non-Rhizopus, -Mucor, and -Lichtheimia Species

PubMed Central

Gomes, Marisa Z. R.; Lewis, Russell E.; Kontoyiannis, Dimitrios P.

2011-01-01

Summary: Rhizopus, Mucor, and Lichtheimia (formerly Absidia) species are the most common members of the order Mucorales that cause mucormycosis, accounting for 70 to 80% of all cases. In contrast, Cunninghamella, Apophysomyces, Saksenaea, Rhizomucor, Cokeromyces, Actinomucor, and Syncephalastrum species individually are responsible for fewer than 1 to 5% of reported cases of mucormycosis. In this review, we provide an overview of the epidemiology, clinical manifestations, diagnosis of, treatment of, and prognosis for unusual Mucormycetes infections (non-Rhizopus, -Mucor, and -Lichtheimia species). The infections caused by these less frequent members of the order Mucorales frequently differ in their epidemiology, geographic distribution, and disease manifestations. Cunninghamella bertholletiae and Rhizomucor pusillus affect primarily immunocompromised hosts, mostly resulting from spore inhalation, causing pulmonary and disseminated infections with high mortality rates. R. pusillus infections are nosocomial or health care related in a large proportion of cases. While Apophysomyces elegans and Saksenaea vasiformis are occasionally responsible for infections in immunocompromised individuals, most cases are encountered in immunocompetent individuals as a result of trauma, leading to soft tissue infections with relatively low mortality rates. Increased knowledge of the epidemiology and clinical presentations of these unusual Mucormycetes infections may improve early diagnosis and treatment. PMID:21482731

Lipid accumulation by pelletized culture of Mucor circinelloides on corn stover hydrolysate.

PubMed

Reis, Cristiano E R; Zhang, Jianguo; Hu, Bo

2014-09-01

Microbial oil accumulated by fungal cells is a potential feedstock for biodiesel production, and lignocellulosic materials can serve as the carbon source to support the fungal growth. The dilute acid pretreatment of corn stover can effectively break down its lignin structure, and this process generates a hydrolysate containing mostly xylose at very dilute concentration and numerous by-products that may significantly inhibit the cell growth. This study utilized corn stover hydrolysate as the culture media for the growth of Mucor circinelloides. The results showed that Mucor cells formed pellets during the cell growth, which facilitates the cell harvest from dilute solution. The results also showed that the inhibitory effect of furfural, 5-hydroxymethylfurfural (HMF), and acetic acid could be avoided if their concentration was low. In fact, all these by-products may be assimilated as carbon sources for the fungal growth. The results proved the feasibility to reuse the cultural broth water for acid pretreatment and then use for subsequent cell cultivation. The results will have a direct impact on the overall water usage of the process.

Degradation of slime extracellular polymeric substances and inhibited sludge flocs destruction contribute to sludge dewaterability enhancement during fungal treatment of sludge using filamentous fungus Mucor sp. GY-1.

PubMed

Wang, Zhenyu; Zheng, Guanyu; Zhou, Lixiang

2015-09-01

Mechanisms responsible for the sludge dewaterability enhanced by filamentous fungi during fungal treatment of sludge were investigated in the present study. The filamentous fungus Mucor sp. GY-1, isolated from waste activated sludge, enhanced sludge dewaterability by 82.1% to achieve the lowest value of normalized sludge specific resistance to filtration (SRF), 8.18 × 10(10) m · L/kg · g-TSS. During the fungal treatment of sludge, 57.8% of slime extracellular polymeric substances (EPS) and 51.1% of polysaccharide in slime EPS were degraded, respectively, by Mucor sp. GY-1, contributing to the improvement of sludge dewaterability. Slime EPS is much more available for Mucor sp. GY-1 than either LB-EPS or TB-EPS that bound with microbial cells. In addition, filamentous fungus Mucor sp. GY-1 entrapped small sludge particles and inhibited the destruction of sludge flocs larger than 100 μm, thus enhancing sludge dewaterability, during fungal treatment of sludge using Mucor sp. GY-1. Copyright © 2015 Elsevier Ltd. All rights reserved.

Mucor irregularis infection around the inner canthus cured by amphotericin B: a case report and review of published literatures.

PubMed

Kang, Daoxian; Jiang, Xian; Wan, Huiying; Ran, Yuping; Hao, Dan; Zhang, Chaoliang

2014-08-01

We report a case of primary cutaneous mucormycosis caused by Mucor irregularis. A 47-year-old farmer was presented to our clinic with the history of progressive red plaque around the inner canthus following dacryocystectomy about a year earlier. Linear, aseptate hyphae were seen by direct KOH examination and in biopsy. Fungal culture revealed light yellow filamentous colonies that were identified as Mucor irregularis by nucleotide sequencing of rRNA gene. Amphotericin B and dexamethasone were used in gradually increasing dosage. The treatment lasted 43 days, and the patient received 760 mg total amphotericin B. The patient was discharged after 2 months of treatment. The plaque became smooth, and fungal culture was negative. There was no recurrence for half a year through telephone follow-ups. A review of published studies revealed 23 cases of Mucor irregularis infection. Most cases resulted following injuries or surgical complications. Farmers and manual laborers were most at risk with males outnumbering females among patients. Amphotericin B and its liposomal preparations remain most effective treatment choices.

Heterothallic mating in Mucor irregularis and first isolate of the species outside of Asia

USDA-ARS?s Scientific Manuscript database

This study reports on the discovery of heterothallic mating in Mucor irregularis (formerly Rhizomucor variabilis var. variabilis) and it extends the range of this species from Asia to the United States. We report on a case of primary cutaneous mucormycosis, involving the forearms of a cotton farmer ...

RNAi pathways in Mucor: A tale of proteins, small RNAs and functional diversity.

PubMed

Torres-Martínez, Santiago; Ruiz-Vázquez, Rosa M

2016-05-01

The existence of an RNA-mediated silencing mechanism in the opportunistic fungal pathogen Mucor circinelloides was first described in the early 2000. Since then, Mucor has reached an outstanding position within the fungal kingdom as a model system to achieve a deeper understanding of regulation of endogenous functions by the RNA interference (RNAi) machinery. M. circinelloides combines diverse components of its RNAi machinery to carry out functions not only limited to the defense against invasive nucleic acids, but also to regulate expression of its own genes by producing different classes of endogenous small RNA molecules (esRNAs). The recent discovery of a novel RNase that participates in a new RNA degradation pathway adds more elements to the gene silencing-mediated regulation. This review focuses on esRNAs in M. circinelloides, the different pathways involved in their biogenesis, and their roles in regulating specific physiological and developmental processes in response to environmental signals, highlighting the complexity of silencing-mediated regulation in fungi. Copyright © 2015 Elsevier Inc. All rights reserved.

Analysis of a Food-Borne Fungal Pathogen Outbreak: Virulence and Genome of a Mucor circinelloides Isolate from Yogurt

PubMed Central

Billmyre, R. Blake; Li, Alicia; Carson, Sandra; Sykes, Sean M.; Huh, Eun Young; Mieczkowski, Piotr; Ko, Dennis C.; Cuomo, Christina A.

2014-01-01

ABSTRACT Food-borne pathogens are ongoing problems, and new pathogens are emerging. The impact of fungi, however, is largely underestimated. Recently, commercial yogurts contaminated with Mucor circinelloides were sold, and >200 consumers became ill with nausea, vomiting, and diarrhea. Mucoralean fungi cause the fatal fungal infection mucormycosis, whose incidence has been continuously increasing. In this study, we isolated an M. circinelloides strain from a yogurt container, and multilocus sequence typing identified the strain as Mucor circinelloides f. circinelloides. M. circinelloides f. circinelloides is the most virulent M. circinelloides subspecies and is commonly associated with human infections, whereas M. circinelloides f. lusitanicus and M. circinelloides f. griseocyanus are less common causes of infection. Whole-genome analysis of the yogurt isolate confirmed it as being close to the M. circinelloides f. circinelloides subgroup, with a higher percentage of divergence with the M. circinelloides f. lusitanicus subgroup. In mating assays, the yogurt isolate formed sexual zygospores with the (−) M. circinelloides f. circinelloides tester strain, which is congruent with its sex locus encoding SexP, the (+) mating type sex determinant. The yogurt isolate was virulent in murine and wax moth larva host systems. In a murine gastromucormycosis model, Mucor was recovered from fecal samples of infected mice for up to 10 days, indicating that Mucor can survive transit through the GI tract. In interactions with human immune cells, M. circinelloides f. lusitanicus induced proinflammatory cytokines but M. circinelloides f. circinelloides did not, which may explain the different levels of virulence in mammalian hosts. This study demonstrates that M. circinelloides can spoil food products and cause gastrointestinal illness in consumers and may pose a particular risk to immunocompromised patients. PMID:25006230

Hematogenously Disseminated Skin Disease Caused by Mucor velutinosus in a Patient with Acute Myeloid Leukemia▿

PubMed Central

Sugui, Janyce A.; Christensen, Jesica A.; Bennett, John E.; Zelazny, Adrian M.; Kwon-Chung, Kyung J.

2011-01-01

We report here a case of disseminated skin infection caused by Mucor velutinosus, a recently described new species. We believe this to be the first published report of a clinical case of mucormycosis due to M. velutinosus, as well as a rare case of dissemination from a deep site to skin. PMID:21543575

Fourier transform infrared spectroscopy for the prediction of fatty acid profiles in Mucor fungi grown in media with different carbon sources.

PubMed

Shapaval, Volha; Afseth, Nils Kristian; Vogt, Gjermund; Kohler, Achim

2014-09-11

Fungal production of polyunsaturated fatty acids (PUFAs) is a highly potential approach in biotechnology. Currently the main focus is directed towards screening of hundreds strains in order to select of few potential ones. Thus, a reliable method for screening a high number of strains within a short period of time is needed. Here, we present a novel method for screening of PUFA-producing fungi by high-throughput microcultivation and FTIR spectroscopy. In the study selected Mucor fungi were grown in media with different carbon sources and fatty acid profiles were predicted on the basis of the obtained spectral data. FTIR spectra were calibrated against fatty acid analysis by GC-FD. The calibration models were cross-validated and correlation coefficients (R2) from 0.71 to 0.78 with RMSECV (root mean squared error) from 2.86% to 6.96% (percentage of total fat) were obtained. The FTIR results show a strong correlation to the results obtained by GC analysis, where high total contents of unsaturated fatty acids (both PUFA and MUFA) were achieved for Mucor plumbeus VI02019 cultivated in canola, olive and sunflower oil and Mucor hiemalis VI01993 cultivated in canola and olive oil.

Sporangiospore size dimorphism is linked to virulence of Mucor circinelloides.

PubMed

Li, Charles H; Cervantes, Maria; Springer, Deborah J; Boekhout, Teun; Ruiz-Vazquez, Rosa M; Torres-Martinez, Santiago R; Heitman, Joseph; Lee, Soo Chan

2011-06-01

Mucor circinelloides is a zygomycete fungus and an emerging opportunistic pathogen in immunocompromised patients, especially transplant recipients and in some cases otherwise healthy individuals. We have discovered a novel example of size dimorphism linked to virulence. M. circinelloides is a heterothallic fungus: (+) sex allele encodes SexP and (-) sex allele SexM, both of which are HMG domain protein sex determinants. M. circinelloides f. lusitanicus (Mcl) (-) mating type isolates produce larger asexual sporangiospores that are more virulent in the wax moth host compared to (+) isolates that produce smaller less virulent sporangiospores. The larger sporangiospores germinate inside and lyse macrophages, whereas the smaller sporangiospores do not. sexMΔ mutants are sterile and still produce larger virulent sporangiospores, suggesting that either the sex locus is not involved in virulence/spore size or the sexP allele plays an inhibitory role. Phylogenetic analysis supports that at least three extant subspecies populate the M. circinelloides complex in nature: Mcl, M. circinelloides f. griseocyanus, and M. circinelloides f. circinelloides (Mcc). Mcc was found to be more prevalent among clinical Mucor isolates, and more virulent than Mcl in a diabetic murine model in contrast to the wax moth host. The M. circinelloides sex locus encodes an HMG domain protein (SexP for plus and SexM for minus mating types) flanked by genes encoding triose phosphate transporter (TPT) and RNA helicase homologs. The borders of the sex locus between the three subspecies differ: the Mcg sex locus includes the promoters of both the TPT and the RNA helicase genes, whereas the Mcl and Mcc sex locus includes only the TPT gene promoter. Mating between subspecies was restricted compared to mating within subspecies. These findings demonstrate that spore size dimorphism is linked to virulence of M. circinelloides species and that plasticity of the sex locus and adaptations in pathogenicity have

Unguisin F, a new cyclic peptide from the endophytic fungus Mucor irregularis.

PubMed

Akone, Sergi H; Daletos, Georgios; Lin, Wenhan; Proksch, Peter

2016-01-01

The new cyclic heptapeptide unguisin F (1) and the known congener unguisin E (2), were obtained from the endophytic fungus Mucor irregularis, isolated from the medicinal plant Moringa stenopetala, collected in Cameroon. The structure of the new compound was unambiguously determined on the basis of one- and two-dimensional NMR spectroscopy as well as by high-resolution mass spectrometry. The absolute configuration of the amino acid residues of 1 and 2 was determined using Marfey's analysis. Compounds 1 and 2 were evaluated for their antibacterial and antifungal potential, but failed to display significant activities.

In vitro and in vivo activities of posaconazole and amphotericin B in a murine invasive infection by Mucor circinelloides: poor efficacy of posaconazole.

PubMed

Salas, Valentina; Pastor, F Javier; Calvo, Enrique; Alvarez, Eduardo; Sutton, Deanna A; Mayayo, Emilio; Fothergill, Anette W; Rinaldi, Michael G; Guarro, Josep

2012-05-01

The in vitro susceptibility of 17 strains of Mucor circinelloides to amphotericin B and posaconazole was ascertained by using broth microdilution and disk diffusion methods and by determining the minimal fungicidal concentration (MFC). We evaluated the efficacy of posaconazole at 40 mg/kg of body weight/day and amphotericin B at 0.8 mg/kg/day in a neutropenic murine model of disseminated infection by M. circinelloides by using 6 different strains tested previously in vitro. In general, most of the posaconazole MICs were within the range of susceptibility or intermediate susceptibility, while the small inhibition zone diameters (IZDs) were indicative of nonsusceptibility for all isolates tested. The MFCs were ≥ 3 dilutions higher than the corresponding MICs. In contrast, amphotericin B showed good activity against all of the strains tested regardless of the method used. The in vivo studies demonstrated that amphotericin B was effective in prolonging survival and reducing the fungal load. Posaconazole showed poor in vivo efficacy with no correlation with the MIC values. The results suggested that posaconazole should be used with caution in the treatment of infections caused by Mucor circinelloides or by strains of Mucor not identified to the species level.

The Role of Strigolactone in the Cross-Talk Between Arabidopsis thaliana and the Endophytic Fungus Mucor sp.

PubMed Central

Rozpądek, Piotr; Domka, Agnieszka M.; Nosek, Michał; Ważny, Rafał; Jędrzejczyk, Roman J.; Wiciarz, Monika; Turnau, Katarzyna

2018-01-01

Over the last years the role of fungal endophytes in plant biology has been extensively studied. A number of species were shown to positively affect plant growth and fitness, thus attempts have been made to utilize these microorganisms in agriculture and phytoremediation. Plant-fungi symbiosis requires multiple metabolic adjustments of both of the interacting organisms. The mechanisms of these adaptations are mostly unknown, however, plant hormones seem to play a central role in this process. The plant hormone strigolactone (SL) was previously shown to activate hyphae branching of mycorrhizal fungi and to negatively affect pathogenic fungi growth. Its role in the plant–endophytic fungi interaction is unknown. The effect of the synthetic SL analog GR24 on the endophytic fungi Mucor sp. growth, respiration, H2O2 production and the activity of antioxidant enzymes was evaluated. We found fungi colony growth rate was decreased in a GR24 concentration dependent manner. Additionally, the fungi accumulated more H2O2 what was accompanied by an altered activity of antioxidant enzymes. Symbiosis with Mucor sp. positively affected Arabidopsis thaliana growth, but SL was necessary for the establishment of the beneficial interaction. A. thaliana biosynthesis mutants max1 and max4, but not the SL signaling mutant max2 did not develop the beneficial phenotype. The negative growth response was correlated with alterations in SA homeostasis and a significant upregulation of genes encoding selected plant defensins. The fungi were also shown to be able to decompose SL in planta and to downregulate the expression of SL biosynthesis genes. Additionally, we have shown that GR24 treatment with a dose of 1 μM activates the production of SA in A. thaliana. The results presented here provide evidence for a role of SL in the plant–endophyte cross-talk during the mutualistic interaction between Arabidopsis thaliana and Mucor sp. PMID:29615990

Successful treatment of a necrotizing fasciitis patient caused by Mucor indicus with amphotericin B and skin grafting.

PubMed

Luo, Yijin; Zeng, Fanqin; Huang, Xiaowen; Li, Qun; Tan, Guozhen; Xi, Liyan; Lu, Changming; Guo, Qing

2014-04-01

Cutaneous mucormycosis, an uncommon disease caused by Mucorales, predominantly occurs in immunocompromised host. The present case is a primary cutaneous mucormycosis due to Mucor indicus in an immunocompetent individual. It is with the features of necrotizing fasciitis over the right pretibial area. We are presenting this case owing to its rarity and the successful treatment with amphotericin B and skin grafting.

Sporangiospore Size Dimorphism Is Linked to Virulence of Mucor circinelloides

PubMed Central

Li, Charles H.; Cervantes, Maria; Springer, Deborah J.; Boekhout, Teun; Ruiz-Vazquez, Rosa M.; Torres-Martinez, Santiago R.; Heitman, Joseph; Lee, Soo Chan

2011-01-01

Mucor circinelloides is a zygomycete fungus and an emerging opportunistic pathogen in immunocompromised patients, especially transplant recipients and in some cases otherwise healthy individuals. We have discovered a novel example of size dimorphism linked to virulence. M. circinelloides is a heterothallic fungus: (+) sex allele encodes SexP and (−) sex allele SexM, both of which are HMG domain protein sex determinants. M. circinelloides f. lusitanicus (Mcl) (−) mating type isolates produce larger asexual sporangiospores that are more virulent in the wax moth host compared to (+) isolates that produce smaller less virulent sporangiospores. The larger sporangiospores germinate inside and lyse macrophages, whereas the smaller sporangiospores do not. sexMΔ mutants are sterile and still produce larger virulent sporangiospores, suggesting that either the sex locus is not involved in virulence/spore size or the sexP allele plays an inhibitory role. Phylogenetic analysis supports that at least three extant subspecies populate the M. circinelloides complex in nature: Mcl, M. circinelloides f. griseocyanus, and M. circinelloides f. circinelloides (Mcc). Mcc was found to be more prevalent among clinical Mucor isolates, and more virulent than Mcl in a diabetic murine model in contrast to the wax moth host. The M. circinelloides sex locus encodes an HMG domain protein (SexP for plus and SexM for minus mating types) flanked by genes encoding triose phosphate transporter (TPT) and RNA helicase homologs. The borders of the sex locus between the three subspecies differ: the Mcg sex locus includes the promoters of both the TPT and the RNA helicase genes, whereas the Mcl and Mcc sex locus includes only the TPT gene promoter. Mating between subspecies was restricted compared to mating within subspecies. These findings demonstrate that spore size dimorphism is linked to virulence of M. circinelloides species and that plasticity of the sex locus and adaptations in pathogenicity

The RNAi machinery controls distinct responses to environmental signals in the basal fungus Mucor circinelloides.

PubMed

Nicolás, Francisco E; Vila, Ana; Moxon, Simon; Cascales, María D; Torres-Martínez, Santiago; Ruiz-Vázquez, Rosa M; Garre, Victoriano

2015-03-25

RNA interference (RNAi) is a conserved mechanism of genome defence that can also have a role in the regulation of endogenous functions through endogenous small RNAs (esRNAs). In fungi, knowledge of the functions regulated by esRNAs has been hampered by lack of clear phenotypes in most mutants affected in the RNAi machinery. Mutants of Mucor circinelloides affected in RNAi genes show defects in physiological and developmental processes, thus making Mucor an outstanding fungal model for studying endogenous functions regulated by RNAi. Some classes of Mucor esRNAs map to exons (ex-siRNAs) and regulate expression of the genes from which they derive. To have a broad picture of genes regulated by the silencing machinery during vegetative growth, we have sequenced and compared the mRNA profiles of mutants in the main RNAi genes by using RNA-seq. In addition, we have achieved a more complete phenotypic characterization of silencing mutants. Deletion of any main RNAi gene provoked a deep impact in mRNA accumulation at exponential and stationary growth. Genes showing increased mRNA levels, as expected for direct ex-siRNAs targets, but also genes with decreased expression were detected, suggesting that, most probably, the initial ex-siRNA targets regulate the expression of other genes, which can be up- or down-regulated. Expression of 50% of the genes was dependent on more than one RNAi gene in agreement with the existence of several classes of ex-siRNAs produced by different combinations of RNAi proteins. These combinations of proteins have also been involved in the regulation of different cellular processes. Besides genes regulated by the canonical RNAi pathway, this analysis identified processes, such as growth at low pH and sexual interaction that are regulated by a dicer-independent non-canonical RNAi pathway. This work shows that the RNAi pathways play a relevant role in the regulation of a significant number of endogenous genes in M. circinelloides during exponential

The RNAi machinery controls distinct responses to environmental signals in the basal fungus Mucor circinelloides

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nicolas, Francisco E.; Vila, Ana; Moxon, Simon

Here, RNA interference (RNAi) is a conserved mechanism of genome defence that can also have a role in the regulation of endogenous functions through endogenous small RNAs (esRNAs). In fungi, knowledge of the functions regulated by esRNAs has been hampered by lack of clear phenotypes in most mutants affected in the RNAi machinery. Mutants of Mucor circinelloides affected in RNAi genes show defects in physiological and developmental processes, thus making Mucor an outstanding fungal model for studying endogenous functions regulated by RNAi. Some classes of Mucor esRNAs map to exons (ex-siRNAs) and regulate expression of the genes from which theymore » derive. To have a broad picture of genes regulated by the silencing machinery during vegetative growth, we have sequenced and compared the mRNA profiles of mutants in the main RNAi genes by using RNA-seq. In addition, we have achieved a more complete phenotypic characterization of silencing mutants Deletion of any main RNAi gene provoked a deep impact in mRNA accumulation at exponential and stationary growth. Genes showing increased mRNA levels, as expected for direct ex-siRNAs targets, but also genes with decreased expression were detected, suggesting that, most probably, the initial ex-siRNA targets regulate the expression of other genes, which can be up- or down-regulated. Expression of 50% of the genes was dependent on more than one RNAi gene in agreement with the existence of several classes of ex-siRNAs produced by different combinations of RNAi proteins. These combinations of proteins have also been involved in the regulation of different cellular processes. Besides genes regulated by the canonical RNAi pathway, this analysis identified processes, such as growth at low pH and sexual interaction that are regulated by a dicer-independent non-canonical RNAi pathway. In conclusion, this work shows that the RNAi pathways play a relevant role in the regulation of a significant number of endogenous genes in M

The RNAi machinery controls distinct responses to environmental signals in the basal fungus Mucor circinelloides

DOE PAGES

Nicolas, Francisco E.; Vila, Ana; Moxon, Simon; ...

2015-03-25

Here, RNA interference (RNAi) is a conserved mechanism of genome defence that can also have a role in the regulation of endogenous functions through endogenous small RNAs (esRNAs). In fungi, knowledge of the functions regulated by esRNAs has been hampered by lack of clear phenotypes in most mutants affected in the RNAi machinery. Mutants of Mucor circinelloides affected in RNAi genes show defects in physiological and developmental processes, thus making Mucor an outstanding fungal model for studying endogenous functions regulated by RNAi. Some classes of Mucor esRNAs map to exons (ex-siRNAs) and regulate expression of the genes from which theymore » derive. To have a broad picture of genes regulated by the silencing machinery during vegetative growth, we have sequenced and compared the mRNA profiles of mutants in the main RNAi genes by using RNA-seq. In addition, we have achieved a more complete phenotypic characterization of silencing mutants Deletion of any main RNAi gene provoked a deep impact in mRNA accumulation at exponential and stationary growth. Genes showing increased mRNA levels, as expected for direct ex-siRNAs targets, but also genes with decreased expression were detected, suggesting that, most probably, the initial ex-siRNA targets regulate the expression of other genes, which can be up- or down-regulated. Expression of 50% of the genes was dependent on more than one RNAi gene in agreement with the existence of several classes of ex-siRNAs produced by different combinations of RNAi proteins. These combinations of proteins have also been involved in the regulation of different cellular processes. Besides genes regulated by the canonical RNAi pathway, this analysis identified processes, such as growth at low pH and sexual interaction that are regulated by a dicer-independent non-canonical RNAi pathway. In conclusion, this work shows that the RNAi pathways play a relevant role in the regulation of a significant number of endogenous genes in M

Characterization of pterin deaminase from Mucor indicus MTCC 3513

NASA Astrophysics Data System (ADS)

Thandeeswaran, M.; Karthika, P.; Mahendran, R.; Palaniswamy, M.; Angayarkanni, J.

2018-03-01

Pterin deaminase is an amidohydrolase enzyme which hydrolyses pteridines to produce lumazine derivatives and ammonia. Even though the enzyme was shown as early as 1959 for its anticancer efficacy there was a long gap in the communique after that which was in 2013. In our study we have chosen Mucor indicus MTCC 3513 which was a promising strain for production of different industrial products.The pterin deaminase enzyme was harvested and extracellular from M. indicus. The extracellular sample was partially purified by using ethanol precipitation and ion exchange column (Hi-Trap QFF) in Fast Protein Liquid Chromatography. The molecular weight of the purified pterin deaminase enzyme was apparently determined by SDS-PAGE. The purified enzyme was further biochemically characterized. Molecular docking studies with the predicted sequence showed higher binding affinity towards folic acid interaction. The structure of this protein may open the windows for new drug targets for cancer therapy.

Microbial transformation of hederagenin by Cunninghamella echinulate, Mucor subtilissimus, and Pseudomonas oleovorans.

PubMed

Liu, Zhen; Lu, Yan-Hua; Feng, Xu; Zou, Ying-Xin; Diao, Zhuo; Chu, Zhi-Yong

2017-07-01

The pentacyclic triterpenoid hederagenin (1) was subjected to biotransformation by Cunninghamella echinulate CGMCC 3.2000, Mucor subtilissimus CGMCC 3.2454 and Pseudomonas oleovorans CGMCC 1.1641. Three metabolites were obtained. On the basis of nuclear magnetic resonance and high-resolution mass spectral analyses, their structures were characterized as 3β, 23-dihydroxyolean-12-en-28-oic acid 28-O-β-D-glucopyranosyl ester (2), 3β, 15α, 23-trihydroxyolean-12-en-28-oic acid (3), 1β, 3β, 23-trihydroxyolean-12-en-28-oic acid (4), and metabolite (3) was a new compound. This was the first report on the biotransformation of hederagenin.

Analysis of a food-borne fungal pathogen outbreak: virulence and genome of a Mucor circinelloides isolate from yogurt.

PubMed

Lee, Soo Chan; Billmyre, R Blake; Li, Alicia; Carson, Sandra; Sykes, Sean M; Huh, Eun Young; Mieczkowski, Piotr; Ko, Dennis C; Cuomo, Christina A; Heitman, Joseph

2014-07-08

Food-borne pathogens are ongoing problems, and new pathogens are emerging. The impact of fungi, however, is largely underestimated. Recently, commercial yogurts contaminated with Mucor circinelloides were sold, and >200 consumers became ill with nausea, vomiting, and diarrhea. Mucoralean fungi cause the fatal fungal infection mucormycosis, whose incidence has been continuously increasing. In this study, we isolated an M. circinelloides strain from a yogurt container, and multilocus sequence typing identified the strain as Mucor circinelloides f. circinelloides. M. circinelloides f. circinelloides is the most virulent M. circinelloides subspecies and is commonly associated with human infections, whereas M. circinelloides f. lusitanicus and M. circinelloides f. griseocyanus are less common causes of infection. Whole-genome analysis of the yogurt isolate confirmed it as being close to the M. circinelloides f. circinelloides subgroup, with a higher percentage of divergence with the M. circinelloides f. lusitanicus subgroup. In mating assays, the yogurt isolate formed sexual zygospores with the (-) M. circinelloides f. circinelloides tester strain, which is congruent with its sex locus encoding SexP, the (+) mating type sex determinant. The yogurt isolate was virulent in murine and wax moth larva host systems. In a murine gastromucormycosis model, Mucor was recovered from fecal samples of infected mice for up to 10 days, indicating that Mucor can survive transit through the GI tract. In interactions with human immune cells, M. circinelloides f. lusitanicus induced proinflammatory cytokines but M. circinelloides f. circinelloides did not, which may explain the different levels of virulence in mammalian hosts. This study demonstrates that M. circinelloides can spoil food products and cause gastrointestinal illness in consumers and may pose a particular risk to immunocompromised patients. Importance: The U.S. FDA reported that yogurt products were contaminated with M

Calcineurin plays key roles in the dimorphic transition and virulence of the human pathogenic zygomycete Mucor circinelloides.

PubMed

Lee, Soo Chan; Li, Alicia; Calo, Silvia; Heitman, Joseph

2013-01-01

Many pathogenic fungi are dimorphic and switch between yeast and filamentous states. This switch alters host-microbe interactions and is critical for pathogenicity. However, in zygomycetes, whether dimorphism contributes to virulence is a central unanswered question. The pathogenic zygomycete Mucor circinelloides exhibits hyphal growth in aerobic conditions but switches to multi-budded yeast growth under anaerobic/high CO₂ conditions. We found that in the presence of the calcineurin inhibitor FK506, Mucor exhibits exclusively multi-budded yeast growth. We also found that M. circinelloides encodes three calcineurin catalytic A subunits (CnaA, CnaB, and CnaC) and one calcineurin regulatory B subunit (CnbR). Mutations in the latch region of CnbR and in the FKBP12-FK506 binding domain of CnaA result in hyphal growth of Mucor in the presence of FK506. Disruption of the cnbR gene encoding the sole calcineurin B subunit necessary for calcineurin activity yielded mutants locked in permanent yeast phase growth. These findings reveal that the calcineurin pathway plays key roles in the dimorphic transition from yeast to hyphae. The cnbR yeast-locked mutants are less virulent than the wild-type strain in a heterologous host system, providing evidence that hyphae or the yeast-hyphal transition are linked to virulence. Protein kinase A activity (PKA) is elevated during yeast growth under anaerobic conditions, in the presence of FK506, or in the yeast-locked cnbR mutants, suggesting a novel connection between PKA and calcineurin. cnaA mutants lacking the CnaA catalytic subunit are hypersensitive to calcineurin inhibitors, display a hyphal polarity defect, and produce a mixture of yeast and hyphae in aerobic culture. The cnaA mutants also produce spores that are larger than wild-type, and spore size is correlated with virulence potential. Our results demonstrate that the calcineurin pathway orchestrates the yeast-hyphal and spore size dimorphic transitions that contribute to

Dual disseminated infection with Nocardia farcinica and Mucor in a patient with systemic lupus erythematosus: a case report.

PubMed

de Clerck, Frederik; Van Ryckeghem, Florence; Depuydt, Pieter; Benoit, Dominque; Druwé, Patrick; Hugel, Arnika; Claeys, Geert; Cools, Piet; Decruyenaere, Johan

2014-11-20

Infections remain a major cause of morbidity and mortality in immunocompromised patients and require early diagnosis and treatment. However, correct diagnosis and treatment are often delayed by a multitude of factors. We report what we believe to be the first case of a combined disseminated infection with Nocardia and Mucor in a patient with systemic lupus erythematosus. A 74-year-old Caucasian woman with systemic lupus erythematosus presented with recurrent pneumonia. Despite empirical treatment with antibiotics, her condition gradually deteriorated. Microbiological sampling by thoracoscopy revealed the presence of Nocardia. Despite the institution of therapy for disseminated nocardiosis, she died of multi-organ failure. A post-mortem investigation confirmed nocardiosis, but showed concomitant disseminated mucormycosis infection as well. Members of the bacterial genus Nocardia and the fungal genus Mucor are ubiquitous in the environment, have the ability to spread to virtually any organ, and are remarkably resistant to appropriate therapy. Both pathogens can mimic other pathologies both on clinical and radiological investigations. Invasive sampling procedures are often needed to prove their presence. Establishing a timely, correct diagnosis and a specific treatment is essential for patient survival.

Complete NMR assignment of a bisecting hybrid-type oligosaccharide transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase.

PubMed

Yamanoi, Takashi; Oda, Yoshiki; Katsuraya, Kaname; Inazu, Toshiyuki; Yamamoto, Kenji

2016-06-02

This study describes the complete nuclear magnetic resonance (NMR) spectral assignment of a bisecting hybrid-type oligosaccharide 1, transferred by Mucor hiemalis endo-β-N-acetylglucosaminidase (Endo-M). Through (1)H- and (13)C-NMR, DQF-COSY, HSQC, HMBC, TOCSY, and NOESY experiments, we determine the structure of the glycoside linkage formed by the Endo-M transglycosylation, i.e., the connection between GlcNAc and GlcNAc in oligosaccharide 1. Copyright © 2016 Elsevier Ltd. All rights reserved.

Multiple isoforms for the catalytic subunit of PKA in the basal fungal lineage Mucor circinelloides.

PubMed

Fernández Núñez, Lucas; Ocampo, Josefina; Gottlieb, Alexandra M; Rossi, Silvia; Moreno, Silvia

2016-12-01

Protein kinase A (PKA) activity is involved in dimorphism of the basal fungal lineage Mucor. From the recently sequenced genome of Mucor circinelloides we could predict ten catalytic subunits of PKA. From sequence alignment and structural prediction we conclude that the catalytic core of the isoforms is conserved, and the difference between them resides in their amino termini. This high number of isoforms is maintained in the subdivision Mucoromycotina. Each paralogue, when compared to the ones form other fungi is more homologous to one of its orthologs than to its paralogs. All of these fungal isoforms cannot be included in the class I or II in which fungal protein kinases have been classified. mRNA levels for each isoform were measured during aerobic and anaerobic growth. The expression of each isoform is differential and associated to a particular growth stage. We reanalyzed the sequence of PKAC (GI 20218944), the only cloned sequence available until now for a catalytic subunit of M. circinelloides. PKAC cannot be classified as a PKA because of its difference in the conserved C-tail; it shares with PKB a conserved C2 domain in the N-terminus. No catalytic activity could be measured for this protein nor predicted bioinformatically. It can thus be classified as a pseudokinase. Its importance can not be underestimated since it is expressed at the mRNA level in different stages of growth, and its deletion is lethal. Copyright © 2016 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Potential of chitosan from Mucor rouxxi UCP064 as alternative natural compound to inhibit Listeria monocytogenes.

PubMed

Bento, Roberta A; Stamford, Tânia L M; de Campos-Takaki, Galba M; Stamford, Thayza C M; de Souza, Evandro L

2009-07-01

Listeria monocytogenes is widely distributed in nature and the infection listeriosis is recognized as a potential threat for human health because of its mortality rate. The objective of this study was to evaluate the growth profile and chitosan production by Mucor rouxxi UCP 064 grown in yam bean (Pachyrhizus erosus L. Urban) medium. It was also to assess the anti-L. monocytogenes efficacy of the obtained chitosan. Higher values of biomass of M. rouxxi (16.9 g.L(-1)) and best yield of chitosan (62 mg.g(-1)) were found after 48 h of cultivation. Residual glucose and nitrogen in the growth media were 4.1 and 0.02 g.L(-1) after 96 h, respectively. Obtained chitosan presented 85 % of degree of deacetylation and 2.60 x 10(4) g.mol(-1) of viscosimetric molecular weight. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values of chitosan against L. monocytogenes ATCC 7644 were, respectively, 2.5 and 5.0 mg.mL(-1). At 2.5 and 5.0 mg.mL(-1) chitosan caused cidal effect in a maximum time of 4 h. Bacterial count below 2 log cfu.mL(-1) were found from 2 h onwards and no recovery in bacterial growth was noted in the remainder period. These results show the biotechnological potential of yam bean medium for chitosan production by Mucor rouxxi and support the possible rational use of chitosan from fungi as natural antimicrobial to control L. monocytogenes.

Potential of chitosan from Mucor rouxxi UCP064 as alternative natural compound to inhibit Listeria monocytogenes

PubMed Central

Bento, Roberta A.; Stamford, Tânia L.M.; de Campos-Takaki, Galba M.; Stamford, Thayza C.M.; de Souza, Evandro L.

2009-01-01

Listeria monocytogenes is widely distributed in nature and the infection listeriosis is recognized as a potential threat for human health because of its mortality rate. The objective of this study was to evaluate the growth profile and chitosan production by Mucor rouxxi UCP 064 grown in yam bean (Pachyrhizus erosus L. Urban) medium. It was also to assess the anti-L. monocytogenes efficacy of the obtained chitosan. Higher values of biomass of M. rouxxi (16.9 g.L-1) and best yield of chitosan (62 mg.g-1) were found after 48 h of cultivation. Residual glucose and nitrogen in the growth media were 4.1 and 0.02 g.L-1 after 96 h, respectively. Obtained chitosan presented 85 % of degree of deacetylation and 2.60 x 104 g.mol-1 of viscosimetric molecular weight. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values of chitosan against L. monocytogenes ATCC 7644 were, respectively, 2.5 and 5.0 mg.mL-1. At 2.5 and 5.0 mg.mL-1 chitosan caused cidal effect in a maximum time of 4 h. Bacterial count below 2 log cfu.mL-1 were found from 2 h onwards and no recovery in bacterial growth was noted in the remainder period. These results show the biotechnological potential of yam bean medium for chitosan production by Mucor rouxxi and support the possible rational use of chitosan from fungi as natural antimicrobial to control L. monocytogenes. PMID:24031403

Evaluation of Mucor indicus and Saccharomyces cerevisiae capability to ferment hydrolysates of rape straw and Miscanthus giganteus as affected by the pretreatment method.

PubMed

Lewandowska, Małgorzata; Szymańska, Karolina; Kordala, Natalia; Dąbrowska, Aneta; Bednarski, Włodzimierz; Juszczuk, Andrzej

2016-07-01

Rape straw and Miscanthus giganteus was pretreated chemically with oxalic acid or sodium hydroxide. The pretreated substrates were hydrolyzed with enzymatic preparations of cellulase, xylanase and cellobiase. The highest concentration of reducing sugars was achieved after hydrolysis of M. giganteus pretreated with NaOH (51.53gdm(-3)). In turn, the highest yield of enzymatic hydrolysis determined based on polysaccharides content in the pretreated substrates was obtained in the experiments with M. giganteus and oxalic acid (99.3%). Rape straw and M. giganteus hydrolysates were fermented using yeast Saccharomyces cerevisiae 7, NRRL 978 or filamentous fungus Mucor rouxii (Mucor indicus) DSM 1191. The highest ethanol concentration was determined after fermentation of M. giganteus hydrolysate pretreated with NaOH using S. cerevisiae (1.92% v/v). Considering cellulose content in the pretreated solid, the highest degree of its conversion to ethanol (86.2%) was achieved after fermentation of the hydrolysate of acid-treated M. giganteus using S. cerevisiae. Copyright © 2016 Elsevier Ltd. All rights reserved.

Quantifying Fungal Viability in Air and Water Samples using Quantitative PCR after Treatment with Propidium Monoazide (PMA)

EPA Science Inventory

A method is described to discriminate between live and dead cells of the infectious fungi Aspergillus fumigatus, A. flavus, A. terreus, Mucor racemosus, Rhizopus stolonifer and Paecilomyces variotii. To test the method, conidial suspensions were heat inactivated at 85oC or held ...

Primary cutaneous mucormycosis caused by Mucor irregularis in an elderly person.

PubMed

Yamaguchi, Sayaka; Okubo, Yuko; Katano, Azusa; Sano, Ayako; Uezato, Hiroshi; Takahashi, Kenzo

2015-02-01

Primary cutaneous mucormycosis is a rare but often lethal severe fungal infection, which usually occurs in immunocompromised patients. We report a case of primary cutaneous mucormycosis caused by Mucor irregularis in an elderly patient. Seven months after the surgical dissection of the involved skin, cutaneous mucormycosis recurred at the peripheral edge of the skin graft. Shortly subsequent to the administration of liposomal amphotericin B, the remaining skin lesion was excised again. M. irregularis is rarely but increasingly reported as a cause of mucormycosis in immunocompetent individuals, especially in Asian farmers. M. irregularis may be largely disseminated in the soils of Asia and thus the trivial trauma at the time of farm work may be a trigger for the onset. These cases tend to leave severe cosmetic damage even in healthy individuals, although the vital prognosis is not affected. © 2014 Japanese Dermatological Association.

Effect of temperature, pH, and water activity on Mucor spp. growth on synthetic medium, cheese analog and cheese.

PubMed

Morin-Sardin, Stéphanie; Rigalma, Karim; Coroller, Louis; Jany, Jean-Luc; Coton, Emmanuel

2016-06-01

The Mucor genus includes a large number of ubiquitous fungal species. In the dairy environment, some of them play a technological role providing typical organoleptic qualities to some cheeses while others can cause spoilage. In this study, we compared the effect of relevant abiotic factors for cheese production on the growth of six strains representative of dairy technological and contaminant species as well as of a non cheese related strain (plant endophyte). Growth kinetics were determined for each strain in function of temperature, water activity and pH on synthetic Potato Dextrose Agar (PDA), and secondary models were fitted to calculate the corresponding specific cardinal values. Using these values and growth kinetics acquired at 15 °C on cheese agar medium (CA) along with three different cheese types, optimal growth rates (μopt) were estimated and consequently used to establish a predictive model. Contrarily to contaminant strains, technological strains showed higher μopt on cheese matrices than on PDA. Interestingly, lag times of the endophyte strain were strongly extended on cheese related matrices. This study offers a relevant predictive model of growth that may be used for better cheese production control but also raises the question of adaptation of some Mucor strains to the cheese. Copyright © 2015 Elsevier Ltd. All rights reserved.

Statistical analysis and modeling of pelletized cultivation of Mucor circinelloides for microbial lipid accumulation.

PubMed

Xia, Chunjie; Wei, Wei; Hu, Bo

2014-04-01

Microbial oil accumulation via oleaginous fungi has some potential benefits because filamentous fungi can form pellets during cell growth and these pellets are easier to harvest from the culture broth than individual cells. This research studied the effect of various culture conditions on the pelletized cell growth of Mucor circinelloides and its lipid accumulation. The results showed that cell pelletization was positively correlated to biomass accumulation; however, pellet size was negatively correlated to the oil content of the fungal biomass, possibly due to the mass transfer barriers generated by the pellet structure. How to control the size of the pellet is the key to the success of the pelletized microbial oil accumulation process.

Detoxification of Fusaric Acid by the Soil Microbe Mucor rouxii.

PubMed

Crutcher, Frankie K; Puckhaber, Lorraine S; Bell, Alois A; Liu, Jinggao; Duke, Sara E; Stipanovic, Robert D; Nichols, Robert L

2017-06-21

Fusarium oxysporum f. sp. vasinfectum race 4 (VCG0114), which causes root rot and wilt of cotton (Gossypium hirsutum and G. barbadense), has been identified recently for the first time in the western hemisphere in certain fields in the San Joaquin Valley of California. This pathotype produces copious quantities of the plant toxin fusaric acid (5-butyl-2-pyridinecarboxylic acid) compared to other isolates of F. oxysporum f. sp. vasinfectum (Fov) that are indigenous to the United States. Fusaric acid is toxic to cotton plants and may help the pathogen compete with other microbes in the soil. We found that a laboratory strain of the fungus Mucor rouxii converts fusaric acid into a newly identified compound, 8-hydroxyfusaric acid. The latter compound is significantly less phytotoxic to cotton than the parent compound. On the basis of bioassays of hydroxylated analogues of fusaric acid, hydroxylation of the butyl side chain of fusaric acid may affect a general detoxification of fusaric acid. Genes that control this hydroxylation may be useful in developing biocontrol agents to manage Fov.

Antioxidants Potential of the Filamentous Fungi (Mucor circinelloides).

PubMed

Hameed, Ahsan; Hussain, Syed Ammar; Yang, Junhuan; Ijaz, Muhammad Umair; Liu, Qing; Suleria, Hafiz Ansar Rasul; Song, Yuanda

2017-10-07

Three important strains of Mucor circinelloides grown in complete and minimal media for specified period (72 h, 120 h and 168 h) under submerged fermentation conditions were investigated for their potential antioxidants/secondary metabolite production. All mycelial extracts demonstrated effective antioxidant activities in terms of β-carotene/linoleic acid bleaching, radical scavenging, reduction of metal ions and chelating abilities against ferrous ions. Different extraction methods and solvent systems affected the recovery yield and antioxidant activities of the extracts significantly ( p ≤ 0.05). Ethanolic extracts were found to be rich source of antioxidant components and subsequently more effective in antioxidant properties. Fermentation period and media used also significantly affected ( p ≤ 0.05) the antioxidant production and the resulting antioxidant properties. The (ethanolic) extracts of all the strains from late exponential growth phase (120 h) showed highest antioxidant production with topmost reducing, chelating and radical scavenging capabilities. Strain MC277.49 was found to be the highest producer of antioxidants followed by MC108.16 and WJ11. Phenolic compounds were detected significantly in higher ( p ≤ 0.05) amount succeeded by the condensed tannins and flavonoids. Total phenol content of each extract was attributed to overall antioxidant capacity. Submerged fermentation with nutritional stress conditions were found to be excellent way of producing surplus amount of natural antioxidants/secondary metabolites with their vast potential commercial application in food and pharmaceutical industries.

Functional identification and regulatory analysis of Δ6-fatty acid desaturase from the oleaginous fungus Mucor sp. EIM-10.

PubMed

Jiang, Xianzhang; Liu, Hongjiao; Niu, Yongchao; Qi, Feng; Zhang, Mingliang; Huang, Jianzhong

2017-03-01

To enlarge the diversity of the desaturases associated with PUFA biosynthesis and to better understand the transcriptional regulation of desaturases, a Δ 6 -desaturase gene (Md6) from Mucor sp. and its 5'-upstream sequence was functionally identified in Saccharomyces cerevisiae. Expression of the Δ 6 -fatty acid desaturase (Md6) in S. cerevisiae showed that Md6 could convert linolenic acid to γ-linolenic acid. Computational analysis of the promoter of Md6 suggested it contains several eukaryotic fundamental transcription regulatory elements. In vivo functional analysis of the promoter showed the 5'-upstream sequence of Md6 could initiate expression of GFP and Md6 itself in S. cerevisiae. A series deletion analysis of the promoter suggested that sequence between -919 to -784 bp (relative to start site) named as eMd6 is the key factor for high activity of Δ 6 -desaturase. The activity of Δ 6 -desaturase was increased by 2.8-fold and 2.5-fold when the eMd6 sequence was placed upstream of -434 with forward or reverse orientations respectively. To our best knowledge, the native promoter of Md6 from Mucor is the strongest promoter for Δ 6 -desaturase reported so far and the sequence between -919 to -784 bp is an enhancer for Δ 6 -desaturase activity.

Characterization and control of Mucor circinelloides spoilage in yogurt.

PubMed

Snyder, Abigail B; Churey, John J; Worobo, Randy W

2016-07-02

Consumer confidence in the food industry is severely affected by large-scale spoilage incidents. However, relatively little research exists on spoilage potential of members of the fungal subphylum Mucormycotina (e.g. Mucor), which includes dimorphic spoilage organisms that can switch between a yeast-like and hyphal phase depending on environmental conditions. The presence of Mucor circinelloides in yogurt may not cause spoilage, but growth and subsequent changes in quality (e.g. container bloating) can cause spoilage if not controlled. The purpose of this study was to evaluate the effects on M. circinelloides of pasteurization regimen, natamycin concentrations, and storage temperature in yogurt production, as measured by fungal proliferation and carbon dioxide production. A strain of M. circinelloides isolated from commercially spoiled yogurt showed greater yogurt-spoilage potential than clinical isolates and other industrial strains. D-values and z-values were determined for the spoilage isolate in milk as an evaluation of the fungus' ability to survive pasteurization. Natamycin was added to yogurt at 0, 5, 10, 15, and 20ppm (μg/ml) to determine its ability to inhibit M. circinelloides over the course of month-long challenge studies at 4°C, 15°C, and 25°C. Survivors were recovered on acidified PDA and carbon dioxide levels were recorded. The D-values at 54°C, 56°C, and 58°C for hyphae/sporangiospores were (in min) 38.31±0.02, 10.17±0.28, and 1.94±0.53, respectively, which yielded a z-value of 3.09°C. The D-values at 51°C, 53°C, and 55°C for yeast-like cells were (in min) 14.25±0.12, 6.87±1.19, and 2.44±0.35, respectively, which yielded a z-value of 0.34°C. These results indicated that M. circinelloides would not survive fluid milk pasteurization if contamination occurred prior to thermal treatment. CO2 production was only observed when M. circinelloides was incubated under low-oxygen conditions, and occurred only at temperatures above 4�

Antioxidants Potential of the Filamentous Fungi (Mucor circinelloides)

PubMed Central

Hussain, Syed Ammar; Yang, Junhuan; Ijaz, Muhammad Umair; Liu, Qing; Song, Yuanda

2017-01-01

Three important strains of Mucor circinelloides grown in complete and minimal media for specified period (72 h, 120 h and 168 h) under submerged fermentation conditions were investigated for their potential antioxidants/secondary metabolite production. All mycelial extracts demonstrated effective antioxidant activities in terms of β-carotene/linoleic acid bleaching, radical scavenging, reduction of metal ions and chelating abilities against ferrous ions. Different extraction methods and solvent systems affected the recovery yield and antioxidant activities of the extracts significantly (p ≤ 0.05). Ethanolic extracts were found to be rich source of antioxidant components and subsequently more effective in antioxidant properties. Fermentation period and media used also significantly affected (p ≤ 0.05) the antioxidant production and the resulting antioxidant properties. The (ethanolic) extracts of all the strains from late exponential growth phase (120 h) showed highest antioxidant production with topmost reducing, chelating and radical scavenging capabilities. Strain MC277.49 was found to be the highest producer of antioxidants followed by MC108.16 and WJ11. Phenolic compounds were detected significantly in higher (p ≤ 0.05) amount succeeded by the condensed tannins and flavonoids. Total phenol content of each extract was attributed to overall antioxidant capacity. Submerged fermentation with nutritional stress conditions were found to be excellent way of producing surplus amount of natural antioxidants/secondary metabolites with their vast potential commercial application in food and pharmaceutical industries. PMID:28991177

Effect of Naphthalene, Butylated Hydroxytoluene, Dioctyl Phthalate, and Adipic Dioctyl Ester, Chemicals Found in the Nests of the Formosan Subterranean Termite (Isoptera: Rhinotermitidae) on a Saprophytic Mucor sp.

USDA-ARS?s Scientific Manuscript database

Fungi are commonly found associated with termites and their nests. Four chemicals that have been isolated from the nests of the Formosan subterranean termite, Coptotermes formosanus Shiraki, were evaluated to determine their effect on a common nest fungus, a saprophytic Mucor sp. Butylated hydroxyto...

Mucor indicus: biology and industrial application perspectives: a review.

PubMed

Karimi, Keikhosro; Zamani, Akram

2013-01-01

Mucor indicus, one of the most important strains of zygomycetes fungi, has been the subject of several studies since a couple of hundred years ago. This fungus, regarded as a non-pathogenic dimorphic microorganism, is used for production of several beers and foods. Morphology of the fungus can be manipulated and well controlled by changing a number of parameters. Furthermore, M. indicus can grow on a variety of substrates including lignocellulosic hydrolysates which are mixtures of hexoses, pentoses, and different severe fermentation inhibitors. Indeed, high yield ethanol production is among the most important features of this strain. Presence of considerable amounts of chitosan in the cell wall is another important aspect of the fungus. Besides production of ethanol and chitosan, the biomass of this fungus has shown a great potential to be used as a rich nutritional source, e.g. fish feed. The fungus is also among the oleaginous fungi and produces high amounts of polyunsaturated fatty acids particularly γ-linolenic acid. Furthermore, the biomass autolysate has a high potential for yeast extract replacement in fermentation by the fungus. Additionally, the strain has shown promising results in heavy metal removal from wastewaters. This review discusses different aspects of biology and industrial application perspectives of M. indicus. Furthermore, open areas for the future basic and applied levels of research are also presented. Copyright © 2013 Elsevier Inc. All rights reserved.

Enhanced bioremediation of lead-contaminated soil by Solanum nigrum L. with Mucor circinelloides.

PubMed

Sun, Liqun; Cao, Xiufeng; Li, Min; Zhang, Xu; Li, Xinxin; Cui, Zhaojie

2017-04-01

Strain selected from mine tailings in Anshan for Pb bioremediation was characterized at the genetic level by internal transcribed spacer (ITS) sequencing. Results revealed that the strain belongs to Mucor circinelloides. Bioremediation of lead-contaminated soil was conducted using Solanum nigrum L. combined with M. circinelloides. The removal efficacy was in the order microbial/phytoremediation > phytoremediation > microbial remediation > control. The bioremediation rates were 58.6, 47.2, and 40.2% in microbial/phytoremediation, microbial remediation, and phytoremediation groups, respectively. Inoculating soil with M. circinelloides enhanced Pb removal and S. nigrum L. growth. The bioaccumulation factor (BF, 1.43), enrichment factor (EF, 1.56), and translocation factor (TF, 1.35) were higher than unit, suggesting an efficient ability of S. nigrum L. in Pb bioremediation. Soil fertility was increased after bioremediation according to change in enzyme activities. The results indicated that inoculating S. nigrum L. with M. circinelloides enhanced its efficiency for phytoremediation of soil contaminated with Pb.

Identification and Characterization of Diacylglycerol Acyltransferase from Oleaginous Fungus Mucor circinelloides.

PubMed

Zhang, Luning; Zhang, Huaiyuan; Song, Yuanda

2018-01-24

Acyl-CoA:diacylglycerol acyltransferase (DGAT) is a pivotal regulator of triacylglycerol (TAG) synthesis. The oleaginous fungus Mucor circinelloides has four putative DGATs: McDGAT1A, McDGAT1B, McDGAT2A, and McDGAT2B, classified into the DGAT1 and DGAT2 subfamilies, respectively. To identify and characterize DGATs in M. circinelloides, these four genes were expressed in Saccharomyces cerevisiae H1246 (TAG-deficient quadruple mutant), individually. TAG biosynthesis was restored only by the expression of McDGAT2B, and TAG content was significantly higher in the mutants with McDGAT2B expression than in a S. cerevisiae mutant with endogenous DGA1 expression. McDGAT2B prefers saturated fatty acids to monounsaturated fatty acids and has an obvious preference for C18:3 (ω-6) according to the results of substrate preference experiments. Furthermore, only the mRNA expression pattern of McDGAT2B correlated with TAG biosynthesis during a fermentation process. Our experiments strongly indicate that McDGAT2B is crucial for TAG accumulation, suggesting that it may be an essential target for metabolic engineering aimed at increasing lipid content of M. circinelloides.

Production, Purification, and Characterization of Polygalacturonase from Mucor circinelloides ITCC 6025

PubMed Central

Thakur, Akhilesh; Pahwa, Roma; Singh, Smarika; Gupta, Reena

2010-01-01

Mucor circinelloides produced an extracellular polygalacturonase enzyme, the production of which was enhanced when various production parameters were optimized. Maximum polygalacturonase (PGase) activity was obtained in 48 h at 30°C and pH 4.0 with pectin methyl ester (1% w/v) as carbon source and a combination of casein hydrolysate (0.1% w/v) and yeast extract (0.1% w/v) as nitrogen source. The enzyme was purified to homogeneity (13.3-fold) by Sephacryl S-100 gel-filtration chromatography. Its molecular weight was 66 kDa on SDS-PAGE. The enzyme was found to have K m and V max values of 2.2 mM and 4.81 IU/ml at 0.1% to 0.5% (w/v) concentration of the substrate. The addition of phenolic acids (0.05 mM), metal ions such as Mn+2, Co+2, Mg+2, Fe+3, Al+3, Hg+2, and Cu+2, and thiols had inhibitory effect on the enzyme. The enzyme showed maximum activity in the presence of polygalacturonic acid (0.1% w/v) at pH 5.5 and 42°C. PMID:21048861

New isocoumarins from a cold-adapted fungal strain mucor sp. and their developmental toxicity to zebrafish embryos.

PubMed

Feng, Chun-Chi; Chen, Guo-Dong; Zhao, Yan-Qiu; Xin, Sheng-Chang; Li, Song; Tang, Jin-Shan; Li, Xiao-Xia; Hu, Dan; Liu, Xing-Zhong; Gao, Hao

2014-07-01

Three new isocoumarin derivatives, mucorisocoumarins A-C (1-3, resp.), together with seven known compounds, 4-10, were isolated from the cold-adapted fungal strain Mucor sp. (No. XJ07027-5). The structures of the new compounds were identified by detailed IR, MS, and 1D- and 2D-NMR analyses. It was noteworthy that compounds 1, 2, 4, and 5 were successfully resolved by chiral HPLC, indicating that 1-7 should exist as enantiomers. In an embryonic developmental toxicity assay using a zebrafish model, compound 3 produced developmental abnormalities in the zebrafish embryos. This is the first report of isocoumarins with developmental toxicity to zebrafish embryos. Copyright © 2014 Verlag Helvetica Chimica Acta AG, Zürich.

Biotransformation of an africanane sesquiterpene by the fungus Mucor plumbeus.

PubMed

Fraga, Braulio M; Díaz, Carmen E; Amador, Leonardo J; Reina, Matías; López-Rodriguez, Matías; González-Coloma, Azucena

2017-03-01

Biotransformation of 8β-hydroxy-african-4(5)-en-3-one angelate by the fungus Mucor plumbeus afforded as main products 6α,8β-dihydroxy-african-4(5)-en-3-one 8β-angelate and 1α,8β-dihydroxy-african-4(5)-en-3-one 8β-angelate, which had been obtained, together with the substrate, from transformed root cultures of Bethencourtia hermosae. This fact shows that the enzyme system involved in these hydroxylations in both organisms, the fungus and the plant, acts with the same regio- and stereospecificity. In addition another twelve derivatives were isolated in the incubation of the substrate, which were identified as the (2'R,3'R)- and (2'S,3'S)-epoxy derivatives of the substrate and of the 6α- and 1α-hydroxy alcohols, the 8β-(2'R,3'R)- and 8β-(2'S,3'S)-epoxyangelate of 8β,15-dihydroxy-african-4(5)-en-3-one, the hydrolysis product of the substrate, and three isomers of 8β-hydroxy-african-4(5)-en-3-one 2ξ,3ξ-dihydroxy-2-methylbutanoate. The insect antifeedant effects of the pure compounds were tested against chewing and sucking insect species along with their selective cytotoxicity against insect (Sf9) and mammalian (CHO) cell lines. Copyright © 2016 Elsevier Ltd. All rights reserved.

(13)C-metabolic flux analysis of lipid accumulation in the oleaginous fungus Mucor circinelloides.

PubMed

Zhao, Lina; Zhang, Huaiyuan; Wang, Liping; Chen, Haiqin; Chen, Yong Q; Chen, Wei; Song, Yuanda

2015-12-01

The oleaginous fungus Mucor circinelloides is of industrial interest because it can produce high levels of polyunsaturated fatty acid γ-linolenic acid. M. circinelloides CBS 277.49 is able to accumulate less than 15% of cell dry weight as lipids, while M. circinelloides WJ11 can accumulate lipid up to 36%. In order to better understand the mechanisms behind the differential lipid accumulation in these two strains, tracer experiments with (13)C-glucose were performed with the growth of M. circinelloides and subsequent gas chromatography-mass spectrometric detection of (13)C-patterns in proteinogenic amino acids was carried out to identify the metabolic network topology and estimate intracellular fluxes. Our results showed that the high oleaginous strain WJ11 had higher flux of pentose phosphate pathway and malic enzyme, lower flux in tricarboxylic acid cycle, higher flux in glyoxylate cycle and ATP: citrate lyase, together, it might provide more NADPH and substrate acetyl-CoA for fatty acid synthesis. Copyright © 2015 Elsevier Ltd. All rights reserved.

Heterologous expression of the Phycomyces blakesleeanus phytoene dehydrogenase gene (carB) in Mucor circinelloides.

PubMed

Ruiz-Hidalgo, M J; Eslava, A P; Alvarez, M I; Benito, E P

1999-11-01

A phytoene dehydrogenase-deficient mutant of Mucor circinelloides accumulating only phytoene was transformed with the gene encoding the corresponding enzyme (carB gene) of Phycomyces blakesleeanus. Carotenoids derived from phytoene were detected in the transformants showing that the P. blakesleeanus carB gene complements the M. circinelloides carB mutation. These newly formed carotenoids accumulated in low quantities, indicating that functional complementation was poor. carB mRNA molecules correctly transcribed were detected in the transformants, but they represented a small proportion of the total population of carB-derived mRNAs, mostly constituted by truncated transcripts and by transcripts longer than the transcript that is functional in Phycomyces. These results showed that the P. blakesleeanus carB gene was expressed in M. circinelloides and suggested that the poor complementation observed was owing, at least in part, to the lack of specificity in the recognition of the transcription initiation and termination signals of the P. blakesleeanus carB gene by the M. circinelloides transcriptional machinery.

Mucor hiemalis mediated 14α-hydroxylation on steroids: in vivo and in vitro investigations of 14α-hydroxylase activity.

PubMed

Kolet, Swati P; Haldar, Saikat; Niloferjahan, Siddiqui; Thulasiram, Hirekodathakallu V

2014-07-01

Transformation of testosterone and progesterone into synthetically challenging 14α-hydroxy derivatives was achieved by using fungal strain Mucor hiemalis. Prolonged incubation led to the formation of corresponding 6β/7α,14α-dihydroxy metabolites. The position and stereochemistry of newly introduced hydroxyl group was determined by detailed spectroscopic analyses. The time course experiment indicated that fungal strain initiated transformation by hydroxylation at 14α-position followed by at 6β- or 7α-positions. Studies using cell-free extracts suggest that the 14α-hydroxylase activity is NADPH dependent and belongs to the cytochrome P450 family. Copyright © 2014 Elsevier Inc. All rights reserved.

Effect of immobilized polygalacturonase from Mucor circinelloides ITCC-6025 on wine fermentation.

PubMed

Sharma, Sakshi; Hiteshi, Kalpana; Gupta, Reena

2013-01-01

Pectinases are among the most widely distributed enzymes in bacteria, fungi, and plants. Almost all the commercial preparations of pectinases are produced from fungal sources. Mucor circinelloides ITCC-6025 produced polygalacturonase when grown in Riviere's medium containing pectin (methyl ester) as the sole source of carbon. Immobilization of purified polygalacturonase was done on silica gel with 86% efficiency. The enzyme took 60 Min to bind maximally on the support. The immobilized enzyme showed maximum activity at a temperature of 45°C (4.57 U/mg) and pH 5.4. The immobilized enzyme was reused for four cycles as it retained almost 55% of its activity. The immobilized enzyme treatment increased the formation of higher alcohols and phenolics during the course of wine formation from apple and plum juices, whereas there was a decrease in the amount of carbohydrates. The enzyme treatment also resulted in clarification of wine; there was an increase in transmittance at 650 nm (201.78% in the case of apple wine and 223.4% in the case of plum wine) as compared to the control (untreated wine). © 2013 International Union of Biochemistry and Molecular Biology, Inc.

A New Acetylcholinesterase Inhibitor from Green Glycosylation of Trachyloban-19-oic Acid by Mucor plumbeus.

PubMed

Santos, Gabriel F Dos; Takahashi, Jacqueline A

2017-01-01

The in vitro metabolism of a widespread natural product, trachyloban-19-oic acid (1), by the fungal species Mucor plumbeus was studied in a sucrose-yeast liquid medium. Two products were isolated, and their structures were determined by spectroscopic means as 7β-hydroxytrachyloban-19-oic acid (5) and trachyloban-19-O-β-D-glucopyranosyl ester (6). To the best of our knowledge, compound 6 is herein reported by the first time in the literature. These compounds were assayed for acetylcholinesterase inhibition along with some related compounds. Compound 6 showed the highest acetylcholinesterase inhibitory activity at 10000 µg/mL among the tested compounds, a result (92.89%) comparable to the activity of the positive control, galanthamine (94.21%). Therefore, biotransformation of the natural product 1 by M. plumbeus produced a novel compound with potential as a new lead to develop anti-Alzheimer medicines.

Alleviation of heavy metal toxicity and phytostimulation of Brassica campestris L. by endophytic Mucor sp. MHR-7.

PubMed

Zahoor, Mahwish; Irshad, Muhammad; Rahman, Hazir; Qasim, Muhammad; Afridi, Sahib Gul; Qadir, Muhammad; Hussain, Anwar

2017-08-01

Heavy metal (HM) pollution is of great concern in countries like Pakistan where a huge proportion of human population is exposed to it. These toxic metals are making their way from water bodies to soil where it not only interferes with plant growth and development but also initiates serious health issues in human consuming the produce of such soils. Bioremediation is one of the most viable and efficient solution for the problem. Purpose of the current study was to isolate endophytic fungi from plants grown on HM contaminated soil and screen them for their ability to tolerate multiple HM including chromium (Cr 6+ ), manganese (Mn 2+ ), cobalt (Co 2+ ), copper (Cu 2+ ) and zinc (Zn 2+ ). Out of 27 isolated endophytes, only one strain (MHR-7) was selected for multiple heavy metals tolerance. The strain was identified as Mucor sp. by 18S and 28S ribosomal RNA internal transcribed spacer (ITS) 1 and 4 sequence homology. The strain effectively tolerated up to 900µgmL -1 of these heavy metals showing no remarkable effect on its growth. The adverse effect of the heavy metals, measured as reduction of the fungal growth increased with increasing concentration of the metals. The strain was able to remove 60-87% of heavy metals from broth culture when supplied with 300µgmL -1 of these metals. A trend of decline in bioremediation potential of the strain was observed with increasing amount of metals. The strain removed metals by biotransformation and/or accumulation of heavy metal in its hyphae. Application of Mucor sp. MHR-7 locked down HM in tis mycelium thereby making them less available to plant root reducing HM uptake and toxicity in mustard. Besides its bioremediation potential, the strain was also able to produce IAA, ACC deaminase and solubilize phosphate making it excellent phytostimulant fungus. It is concluded that MHR-7 is an excellent candidate for use as biofertilizer in fields affected with heavy metals. Copyright © 2017 Elsevier Inc. All rights reserved.

Rhizovarins A-F, Indole-Diterpenes from the Mangrove-Derived Endophytic Fungus Mucor irregularis QEN-189.

PubMed

Gao, Shu-Shan; Li, Xiao-Ming; Williams, Katherine; Proksch, Peter; Ji, Nai-Yun; Wang, Bin-Gui

2016-08-26

Genome mining of the fungus Mucor irregularis (formerly known as Rhizomucor variabilis) revealed the presence of various gene clusters for secondary metabolite biosynthesis, including several terpene-based clusters. Investigation into the chemical diversity of M. irregularis QEN-189, an endophytic fungus isolated from the fresh inner tissue of the marine mangrove plant Rhizophora stylosa, resulted in the discovery of 20 structurally diverse indole-diterpenes including six new compounds, namely, rhizovarins A-F (1-6). Among them, compounds 1-3 represent the most complex members of the reported indole-diterpenes. The presence of an unusual acetal linked to a hemiketal (1) or a ketal (2 and 3) in an unprecedented 4,6,6,8,5,6,6,6,6-fused indole-diterpene ring system makes them chemically unique. Their structures and absolute configurations were elucidated by spectroscopic analysis, modified Mosher's method, and chemical calculations. Each of the isolated compounds was evaluated for antitumor activity against HL-60 and A-549 cell lines.

Biomineralization of gold by Mucor plumbeus: The progress in understanding the mechanism of nanoparticles' formation.

PubMed

Maliszewska, Irena; Tylus, Włodzimierz; Chęcmanowski, Jacek; Szczygieł, Bogdan; Pawlaczyk-Graja, Izabela; Pusz, Wojciech; Baturo-Cieśniewska, Anna

2017-09-01

This contribution describes the deposition of gold nanoparticles by microbial reduction of Au(III) ions using the mycelium of Mucor plumbeus. Biosorption as the major mechanism of Au(III) ions binding by the fungal cells and the reduction of them to the form of Au(0) on/in the cell wall, followed by the transportation of the synthesized gold nanoparticles to the cytoplasm, is postulated. The probable mechanism behind the reduction of Au(III) ions is discussed, leading to the conclusion that this process is nonenzymatic one. Chitosan of the fungal cell wall is most likely to be the major molecule involved in biomineralization of gold by the mycelium of M. plumbeus. Separation of gold nanoparticles from the cells has been carried out by the ultrasonic disintegration and the obtained nanostructures were characterized by UV-vis spectroscopy and transmission electron micrograph analysis. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1381-1392, 2017. © 2017 American Institute of Chemical Engineers.

Hydroxylation of 1,8-cineole by Mucor ramannianus and Aspergillus niger.

PubMed

Ramos, Aline de Souza; Ribeiro, Joyce Benzaquem; Teixeira, Bruna Gomes; Ferreira, José Luiz Pinto; Silva, Jefferson Rocha de A; Ferreira, Alexandre do Amaral; de Souza, Rodrigo Octavio Mendonça Alves; Amaral, Ana Claudia F

2015-03-01

The monoterpenoid 1,8-cineole is obtained from the leaves of Eucalyptus globulus and it has important biological activities. It is a cheap natural substrate because it is a by-product of the Eucalyptus cultivation for wood and pulp production. In this study, it was evaluated the potential of three filamentous fungi in the biotransformation of 1,8-cineole. The study was divided in two steps: first, reactions were carried out with 1,8-cineole at 1 g/L for 24 h; afterwards, reactions were carried out with substrate at 5 g/L for 5 days. The substrate was hydroxylated into 2-exo-hydroxy-1,8-cineole and 3-exo-hydroxy-1,8-cineole by fungi Mucor ramannianus and Aspergillus niger with high stereoselectivity. Trichoderma harzianum was also tested but no transformation was detected. M. ramannianus led to higher than 99% of conversion within 24 h with a starting high substrate concentration (1 g/L). When substrate was added at 5 g/L, only M. ramannianus was able to catalyze the reaction, but the conversion level was 21.7% after 5 days. Both products have defined stereochemistry and could be used as chiral synthons. Furthermore, biological activity has been described for 3-exo-hydroxy-1,8-cineol. To the best of our knowledge, this is the first report on the use of M. ramannianus in this reaction.

Role of malate transporter in lipid accumulation of oleaginous fungus Mucor circinelloides.

PubMed

Zhao, Lina; Cánovas-Márquez, José T; Tang, Xin; Chen, Haiqin; Chen, Yong Q; Chen, Wei; Garre, Victoriano; Song, Yuanda; Ratledge, Colin

2016-02-01

Fatty acid biosynthesis in oleaginous fungi requires the supply of reducing power, NADPH, and the precursor of fatty acids, acetyl-CoA, which is generated in the cytosol being produced by ATP: citrate lyase which requires citrate to be, transported from the mitochondrion by the citrate/malate/pyruvate transporter. This transporter, which is within the mitochondrial membrane, transports cytosolic malate into the mitochondrion in exchange for mitochondrial citrate moving into the cytosol (Fig. 1). The role of malate transporter in lipid accumulation in oleaginous fungi is not fully understood, however. Therefore, the expression level of the mt gene, coding for a malate transporter, was manipulated in the oleaginous fungus Mucor circinelloides to analyze its effect on lipid accumulation. The results showed that mt overexpression increased the lipid content for about 70 % (from 13 to 22 % dry cell weight, CDW), whereas the lipid content in mt knockout mutant decreased about 27 % (from 13 to 9.5 % CDW) compared with the control strain. Furthermore, the extracellular malate concentration was decreased in the mt overexpressing strain and increased in the mt knockout strain compared with the wild-type strain. This work suggests that the malate transporter plays an important role in regulating lipid accumulation in oleaginous fungus M. circinelloides.

A new regulatory mechanism controlling carotenogenesis in the fungus Mucor circinelloides as a target to generate β-carotene over-producing strains by genetic engineering.

PubMed

Zhang, Yingtong; Navarro, Eusebio; Cánovas-Márquez, José T; Almagro, Lorena; Chen, Haiqin; Chen, Yong Q; Zhang, Hao; Torres-Martínez, Santiago; Chen, Wei; Garre, Victoriano

2016-06-07

Carotenoids are natural pigments with antioxidant properties that have important functions in human physiology and must be supplied through the diet. They also have important industrial applications as food colourants, animal feed additives and nutraceuticals. Some of them, such as β-carotene, are produced on an industrial scale with the use of microorganisms, including fungi. The mucoral Blakeslea trispora is used by the industry to produce β-carotene, although optimisation of production by molecular genetic engineering is unfeasible. However, the phylogenetically closely related Mucor circinelloides, which is also able to accumulate β-carotene, possesses a vast collection of genetic tools with which to manipulate its genome. This work combines classical forward and modern reverse genetic techniques to deepen the regulation of carotenoid synthesis and generate candidate strains for biotechnological production of β-carotene. Mutagenesis followed by screening for mutants with altered colour in the dark and/or in light led to the isolation of 26 mutants that, together with eight previously isolated mutants, have been analysed in this work. Although most of the mutants harboured mutations in known structural and regulatory carotenogenic genes, eight of them lacked mutations in those genes. Whole-genome sequencing of six of these strains revealed the presence of many mutations throughout their genomes, which makes identification of the mutation that produced the phenotype difficult. However, deletion of the crgA gene, a well-known repressor of carotenoid biosynthesis in M. circinelloides, in two mutants (MU206 and MU218) with high levels of β-carotene resulted in a further increase in β-carotene content to differing extents with respect to the crgA single-null strain; in particular, one strain derived from MU218 was able to accumulate up to 4 mg/g of β-carotene. The additive effect of crgA deletion and the mutations present in MU218 suggests the existence of a

Physico-chemical characteristics and functional properties of chitin and chitosan produced by Mucor circinelloides using yam bean as substrate.

PubMed

Fai, Ana Elizabeth C; Stamford, Thayza C M; Stamford-Arnaud, Thatiana M; Santa-Cruz, Petrus D'Amorim; da Silva, Marta C Freitas; Campos-Takaki, Galba M; Stamford, Tânia L M

2011-08-23

Microbiological processes were used for chitin and chitosan production by Mucor circinelloides (UCP 050) grown in yam bean (Pachyrhizus erosus L. Urban) medium. The polysaccharides were extracted by alkali-acid treatment and structural investigations by X-ray diffraction, Fourier transform IR analysis, viscosity and thermal analysis by TG, DTG, and DTA were done. The highest biomass yield (20.7 g/L) was obtained at 96 hours. The highest levels of chitosan (64 mg/g) and chitin (500 mg/g) were produced at 48 and 72 hours, respectively. It was demonstrated that yam bean shows great potential as an economic medium and it is possible to achieve a good yield of chitosan with chemical properties that enable its use in biotechnological applications.

Responses of the antioxidative and biotransformation enzymes in the aquatic fungus Mucor hiemalis exposed to cyanotoxins.

PubMed

Balsano, Evelyn; Esterhuizen-Londt, Maranda; Hoque, Enamul; Lima, Stephan Pflugmacher

2017-08-01

To investigate antioxidative and biotransformation enzyme responses in Mucor hiemalis towards cyanotoxins considering its use in mycoremediation applications. Catalase (CAT), glutathione reductase (GR), and glutathione peroxidase (GPx) in M. hiemalis maintained their activities at all tested microcystin-LR (MC-LR) exposure concentrations. Cytosolic glutathione S-transferase (GST) activity decreased with exposure to 100 µg MC-LR l -1 while microsomal GST remained constant. Cylindrospermopsin (CYN) at 100 µg l -1 led to an increase in CAT activity and inhibition of GR, as well as to a concentration-dependent GPx inhibition. Microsomal GST was inhibited at all concentrations tested. β-N-methylamino-L-alanine (BMAA) inhibited GR activity in a concentration-dependent manner, however, CAT, GPx, and GST remained unaffected. M. hiemalis showed enhanced oxidative stress tolerance and intact biotransformation enzyme activity towards MC-LR and BMAA in comparison to CYN, confirming its applicability in bioreactor technology in terms of viability and survival in their presence.

LC-MS/MS method development for quantitative analysis of acetaminophen uptake by the aquatic fungus Mucor hiemalis.

PubMed

Esterhuizen-Londt, Maranda; Schwartz, Katrin; Balsano, Evelyn; Kühn, Sandra; Pflugmacher, Stephan

2016-06-01

Acetaminophen is a pharmaceutical, frequently found in surface water as a contaminant. Bioremediation, in particular, mycoremediation of acetaminophen is a method to remove this compound from waters. Owing to the lack of quantitative analytical method for acetaminophen in aquatic organisms, the present study aimed to develop a method for the determination of acetaminophen using LC-MS/MS in the aquatic fungus Mucor hiemalis. The method was then applied to evaluate the uptake of acetaminophen by M. hiemalis, cultured in pellet morphology. The method was robust, sensitive and reproducible with a lower limit of quantification of 5 pg acetaminophen on column. It was found that M. hiemalis internalize the pharmaceutical, and bioaccumulate it with time. Therefore, M. hiemalis was deemed a suitable candidate for further studies to elucidate its pharmaceutical tolerance and the longevity in mycoremediation applications. Copyright © 2016 Elsevier Inc. All rights reserved.

New cryptotanshinone derivatives with anti-influenza A virus activities obtained via biotransformation by Mucor rouxii.

PubMed

He, Wenni; Li, Yao; Qin, Yuejie; Tong, Xiaomei; Song, Zhijun; Zhao, Yu; Wei, Ran; Li, Li; Dai, Huanqin; Wang, Wenzhao; Luo, Houwei; Ye, Xin; Zhang, Lixin; Liu, Xueting

2017-08-01

This paper provides an efficient platform to diversify the structure and pharmaceutical potentials of known natural products. Seven metabolites were obtained via the biotransformation of cryptotanshinone by the fungus Mucor rouxii AS 3.3447, and assigned as 13R-14R-hydroxy-anhydride of 16R-cryptotanshinone (1), 1S-hydroxy-anhydride of 16R-cryptotanshinone (2), 1R-hydroxy-anhydride of 16R-cryptotanshinone (3), 3S-hydroxy-epicryptoacetalide (4), 3S-hydroxy-cryptoacetalide (5), epicryptoacetalide (6), and cryptoacetalide (7). Among these compounds, 1-5 are novel. The ortho-naphthoquinone chromophore of cryptotanshinone was degraded and rearranged by M. rouxii. 1 and 3 showed good anti-influenza A virus activities with the reduced cytotoxic activities compared to the parent substrate cryptotanshinone (8). The structures of all the new compounds were determined on the basis of HRESIMS (high-resolution electrospray ionization mass spectroscopy) spectrometry, NMR (nuclear magnetic resonance) spectroscopy, ECD (electronic circular dichroism) calculations, and the CD (circular dichroism) of "in situ" method with [Rh 2 (OCOCF 3 ) 4 ].

Purification, biochemical, and structural characterization of a novel fibrinolytic enzyme from Mucor subtilissimus UCP 1262.

PubMed

Nascimento, Thiago Pajeú; Sales, Amanda Emmanuelle; Porto, Tatiana Souza; Costa, Romero Marcos Pedrosa Brandão; Breydo, Leonid; Uversky, Vladimir N; Porto, Ana Lúcia Figueiredo; Converti, Attilio

2017-08-01

Fibrinolytic proteases are enzymes that degrade fibrin. They provide a promising alternative to existing drugs for thrombolytic therapy. A protease isolated from the filamentous fungus Mucor subtilissimus UCP 1262 was purified in three steps by ammonium sulfate fractionation, ion exchange, and molecular exclusion chromatographies, and characterized biochemically and structurally. The purified protease exhibited a molecular mass of 20 kDa, an apparent isoelectric point of 4.94 and a secondary structure composed mainly of α-helices. Selectivity for N-succinyl-Ala-Ala-Pro-Phe-p-nitroanilide as substrate suggests that this enzyme is a chymotrypsin-like serine protease, whose activity was enhanced by the addition of Cu 2+ , Mg 2+ , and Fe 2+ . The enzyme showed a fibrinolytic activity of 22.53 U/mL at 40 °C and its contact with polyethylene glycol did not lead to any significant alteration of its secondary structure. This protein represents an important example of a novel fibrinolytic enzyme with potential use in the treatment of thromboembolic disorders such as strokes, pulmonary emboli, and deep vein thrombosis.

Genome-scale metabolic modeling of Mucor circinelloides and comparative analysis with other oleaginous species.

PubMed

Vongsangnak, Wanwipa; Klanchui, Amornpan; Tawornsamretkit, Iyarest; Tatiyaborwornchai, Witthawin; Laoteng, Kobkul; Meechai, Asawin

2016-06-01

We present a novel genome-scale metabolic model iWV1213 of Mucor circinelloides, which is an oleaginous fungus for industrial applications. The model contains 1213 genes, 1413 metabolites and 1326 metabolic reactions across different compartments. We demonstrate that iWV1213 is able to accurately predict the growth rates of M. circinelloides on various nutrient sources and culture conditions using Flux Balance Analysis and Phenotypic Phase Plane analysis. Comparative analysis of three oleaginous genome-scale models, including M. circinelloides (iWV1213), Mortierella alpina (iCY1106) and Yarrowia lipolytica (iYL619_PCP) revealed that iWV1213 possesses a higher number of genes involved in carbohydrate, amino acid, and lipid metabolisms that might contribute to its versatility in nutrient utilization. Moreover, the identification of unique and common active reactions among the Zygomycetes oleaginous models using Flux Variability Analysis unveiled a set of gene/enzyme candidates as metabolic engineering targets for cellular improvement. Thus, iWV1213 offers a powerful metabolic engineering tool for multi-level omics analysis, enabling strain optimization as a cell factory platform of lipid-based production. Copyright © 2016 Elsevier B.V. All rights reserved.

Sialoglycoproteins in morphological distinct stages of Mucor polymorphosporus and their influence on phagocytosis by human blood phagocytes.

PubMed

Almeida, Catia Amancio; de Campos-Takaki, Galba Maria; Portela, Maristela Barbosa; Travassos, Luiz R; Alviano, Celuta Sales; Alviano, Daniela Sales

2013-10-01

The possible role of sialic acids in host cells-fungi interaction and their association with glycoproteins were evaluated using a clinical isolate of the dimorphic fungus Mucor polymorphosporus. Lectin-binding assays with spores and yeast cells denoted the presence of surface sialoglycoconjugates containing 2,3- and 2,6-linked sialylglycosyl groups. Western blotting with peroxidase-labeled Limulus polyphemus agglutinin revealed the occurrence of different sialoglycoprotein types in both cell lysates and cell wall protein extracts of mycelia, spores, and yeasts of M. polymorphosporus. Sialic acids contributed to the surface negative charge of spores and yeast forms as evaluated by adherence to a cationic substrate. Sialidase-treated spores were less resistant to phagocytosis by human neutrophils and monocytes from healthy individuals than control (untreated) fungal suspensions. The results suggest that sialic acids are terminal units of various glycoproteins of M. polymorphosporus, contributing to negative charge of yeasts and spore cells and protecting infectious propagules from destruction by host cells.

Dual Functions of Lip6 and Its Regulation of Lipid Metabolism in the Oleaginous Fungus Mucor circinelloides.

PubMed

Zan, Xinyi; Tang, Xin; Chu, Linfang; Song, Yuanda

2018-03-21

Although multiple roles of lipases have been reported in yeasts and microalgae, the functions of lipases have not been studied in oleaginous filamentous fungi. Lipase Lip6 has been reported in the oleaginous filamentous fungus Mucor circinelloides with the consensus lipase motif GXSXG and the typical acyltransferase motif of H-(X) 4 -D. To demonstrate that Lip6 might play dual roles as a lipase and an acyltransferase, we performed site-directed mutagenesis in the lipase motif and the acyltransferase motif of Lip6. Mutation in the lipase motif increased cell biomass by 12%-18% and promoted lipid accumulation by 9%-24%, while mutation in the acyltransferase motif induced lipid degradation. In vitro, purified Lip6 had a slight lipase activity but had a stronger phospholipid:DAG acyltransferase activity. Enzyme activity assays in vivo and phospholipid synthesis pathway analysis suggested that phosphatidyl serine and phosphatidyl ethanolamine can be the supplier of a fatty acyl moiety to form TAG in M. circinelloides.

Morphological changes of the filamentous fungus Mucor mucedo and inhibition of chitin synthase activity induced by anethole.

PubMed

Yutani, Masahiro; Hashimoto, Yukie; Ogita, Akira; Kubo, Isao; Tanaka, Toshio; Fujita, Ken-ichi

2011-11-01

trans-Anethole (anethole), a major component of anise oil, has a broad antimicrobial spectrum with antimicrobial activity relatively weaker than those of well-known antibiotics, and significantly enhances the antifungal activity of polygodial and dodecanol against the baker's yeast Saccharomyces cerevisiae and human pathogenic yeast Candida albicans. However, the antifungal mechanism of anethole is unresolved. Anethole demonstrated antifungal activity against the filamentous fungus, Mucor mucedo IFO 7684, accompanied by hyphal morphological changes such as swollen hyphae at the tips. Its minimum growth inhibitory concentration was 0.625 mM. A hyperosmotic condition (1.2 M sorbitol) restricted the induction of morphological changes, while hypoosmotic treatment (distilled water) induced bursting of hyphal tips and leakage of cytoplasmic constituents. Furthermore, anethole dose-dependently inhibited chitin synthase (CHS) activity in permeabilized hyphae in an uncompetitive manner. These results suggest that the morphological changes of M. mucedo could be explained by the fragility of cell walls caused by CHS inhibition. Copyright © 2011 John Wiley & Sons, Ltd.

Value-added oil and animal feed production from corn-ethanol stillage using the oleaginous fungus Mucor circinelloides.

PubMed

Mitra, Debjani; Rasmussen, Mary L; Chand, Priyanka; Chintareddy, Venkat Reddy; Yao, Linxing; Grewell, David; Verkade, John G; Wang, Tong; van Leeuwen, J Hans

2012-03-01

This study highlights the potential of oleaginous fungus, Mucor circinelloides in adsorbing/assimilating oil and nutrients in thin stillage (TS), and producing lipid and protein-rich fungal biomass. Fungal cultivation on TS for 2 days in a 6-L airlift bioreactor, resulted in a 92% increase in oil yield from TS, and 20 g/L of fungal biomass (dry) with a lipid content of 46% (g of oil per 100g dry biomass). Reduction in suspended solids and soluble chemical oxygen demand (SCOD) in TS were 95% and 89%, respectively. The polyunsaturated fatty acids in fungal oil were 52% of total lipids. Fungal cells grown on Yeast Malt (YM) broth had a higher concentration of γ-linolenic acid (17 wt.%) than those grown on TS (1.4 wt.%). Supplementing TS with crude glycerol (10%, v/v) during the stationary growth phase led to a further 32% increase (from 46% to 61%) in cellular oil content. Copyright © 2011 Elsevier Ltd. All rights reserved.

Lysine and Glutamic Acids as the End Products of Multi-response of Optimized Fermented Medium by Mucor mucedo KP736529.

PubMed

El-Hersh, Mohammed S; Saber, WesamEldin I A; El-Fadaly, Husain A; Mahmoud, Mohammed K

Amino acids are important for living organisms, they acting as crucial for metabolic activities and energy generation, wherein the deficiency in these amino acids cause various physiological defects. The aim of this study is to investigate the effect of some nutritional factors on the amino acids production by Mucor mucedo KP736529 during fermentation intervals. Mucor mucedo KP736529 was selected according to proteolytic activity. Corn steep liquor and olive cake were used in the fermented medium during Placket-Burman and central composite design to maximize the production of lysine and glutamic acids. During the screening by Plackett-Burman design, olive cake and Corn Steep Liquor (CSL) had potential importance for the higher production of amino acids. The individual fractionation of total amino acids showed both lysine and glutamic as the major amino acids associated with the fermentation process. Moreover, the Central Composite Design (CCD) has been adopted to explain the interaction between olive cake and CSL on the production of lysine and glutamic acids. The model recorded significant F-value, with high values of R 2, adjusted R 2 and predicted R 2 for both lysine and glutamic, indicating the validity of the data. Solving equation for maximum production of lysine recorded theoretical levels of olive cake and CSL, being 2.58 and 1.83 g L -1, respectively, with predicting value of lysine at 1.470 μg mL -1, whereas the predicting value of glutamic acid reached 0.805 mg mL -1 at levels of 2.49 and 1.93 g L -1 from olive cake and CSL, respectively. The desirability function (D) showed the actual responses being 1.473±0.009 and 0.801±0.004 μg mL -1 for lysine and glutamic acids, respectively. The model showed adequate validity to be applied in a large-scale production of both lysine and glutamic acids.

Novel glucose dehydrogenase from Mucor prainii: Purification, characterization, molecular cloning and gene expression in Aspergillus sojae.

PubMed

Satake, Ryoko; Ichiyanagi, Atsushi; Ichikawa, Keiichi; Hirokawa, Kozo; Araki, Yasuko; Yoshimura, Taro; Gomi, Keiko

2015-11-01

Glucose dehydrogenase (GDH) is of interest for its potential applications in the field of glucose sensors. To improve the performance of glucose sensors, GDH is required to have strict substrate specificity. A novel flavin adenine dinucleotide (FAD)-dependent GDH was isolated from Mucor prainii NISL0103 and its enzymatic properties were characterized. This FAD-dependent GDH (MpGDH) exhibited high specificity toward glucose. High specificity for glucose was also observed even in the presence of saccharides such as maltose, galactose and xylose. The molecular masses of the glycoforms of GDH ranged from 90 to 130 kDa. After deglycosylation, a single 80 kDa band was observed. The gene encoding MpGDH was cloned and expressed in Aspergillus sojae. The apparent kcat and Km values of recombinant enzyme for glucose were found to be 749.7 s(-1) and 28.3 mM, respectively. The results indicated that the characteristics of MpGDH were suitable for assaying blood glucose levels. Copyright © 2015 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

Phylogenetic analysis reveals two genotypes of the emerging fungus Mucor indicus, an opportunistic human pathogen in immunocompromised patients.

PubMed

Taj-Aldeen, Saad J; Almaslamani, Muna; Theelen, Bart; Boekhout, Teun

2017-07-12

Mucormycosis is a rare fungal infection caused by Mucor indicus. Phylogenetic analysis of many M. indicus isolates, mainly sampled from different clinical and environmental specimens collected worldwide, revealed two genotypes, I and II, based on ITS and D1/D2 LSU rDNA sequences. A retrospective review of the literature revealed 13 cases. Eight (76.9%) patients had disseminated infections, and the overall mortality rate was 30.7%. A pulmonary infection caused by M. indicus genotype I in a liver transplant recipient was disseminated to include the skin and was successfully treated with liposomal amphotericin B and aggressive surgery. M. indicus can infect a wide variety of patients with no real preference for the site of infection. We concluded that M. indicus has emerged as a significant cause of invasive mycosis in severely immunocompromised patients worldwide. Early diagnosis and initiation of appropriate therapy could enhance survival in these immunocompromised patient populations.

Generation of lycopene-overproducing strains of the fungus Mucor circinelloides reveals important aspects of lycopene formation and accumulation.

PubMed

Zhang, Yingtong; Chen, Haiqin; Navarro, Eusebio; López-García, Sergio; Chen, Yong Q; Zhang, Hao; Chen, Wei; Garre, Victoriano

2017-03-01

To generate lycopene-overproducing strains of the fungus Mucor circinelloides with interest for industrial production and to gain insight into the catalytic mechanism of lycopene cyclase and regulatory process during lycopene overaccumulation. Three lycopene-overproducing mutants were generated by classic mutagenesis techniques from a β-carotene-overproducing strain. They carried distinct mutations in the carRP gene encoding lycopene cyclase that produced loss of enzymatic activity to different extents. In one mutant (MU616), the lycopene cyclase was completely destroyed, and a 43.8% (1.1 mg/g dry mass) increase in lycopene production was observed in comparison to that by the previously existing lycopene overproducer. In addition, feedback regulation of the end product was suggested in lycopene-overproducing strains. A lycopene-overaccumulating strain of the fungus M. circinelloides was generated that could be an alternative for the industrial production of lycopene. Vital catalytic residues for lycopene cyclase activity and the potential mechanism of lycopene formation and accumulation were identified.

Obesity changes the human gut mycobiome

PubMed Central

Mar Rodríguez, M.; Pérez, Daniel; Javier Chaves, Felipe; Esteve, Eduardo; Marin-Garcia, Pablo; Xifra, Gemma; Vendrell, Joan; Jové, Mariona; Pamplona, Reinald; Ricart, Wifredo; Portero-Otin, Manuel; Chacón, Matilde R.; Fernández Real, José Manuel

2015-01-01

The human intestine is home to a diverse range of bacterial and fungal species, forming an ecological community that contributes to normal physiology and disease susceptibility. Here, the fungal microbiota (mycobiome) in obese and non-obese subjects was characterized using Internal Transcribed Spacer (ITS)-based sequencing. The results demonstrate that obese patients could be discriminated by their specific fungal composition, which also distinguished metabolically “healthy” from “unhealthy” obesity. Clusters according to genus abundance co-segregated with body fatness, fasting triglycerides and HDL-cholesterol. A preliminary link to metabolites such as hexadecanedioic acid, caproic acid and N-acetyl-L-glutamic acid was also found. Mucor racemosus and M. fuscus were the species more represented in non-obese subjects compared to obese counterparts. Interestingly, the decreased relative abundance of the Mucor genus in obese subjects was reversible upon weight loss. Collectively, these findings suggest that manipulation of gut mycobiome communities might be a novel target in the treatment of obesity. PMID:26455903

Purification and properties of extracellular polysaccharide (EPS) antigens produced by different mould species.

PubMed

Notermans, S; Wieten, G; Engel, H W; Rombouts, F M; Hoogerhout, P; van Boom, J H

1987-02-01

Extracellular polysaccharide (EPS) antigens produced by different mould species were purified and partially characterized. Purification included (NH4)2SO4 treatment, Sepharose CL-4B column chromatography and Con A-sepharose chromatography. The EPS of Penicillium digitatum, Mucor racemosus and Cladosporium cladosporioides showed high antigenic capacities. Immunologically the EPS were partially genus-specific, but cross-reactivity was observed. The EPS antigens produced by species of Penicillium, Aspergillus repens and Geotrichum candidum lost their immunological activity upon heating (100 degrees C) at pH 1.8, while the EPS antigen of M. racemosus, Rhizopus oligosporus and C. cladosporioides were stable under the same conditions. The dominant monosaccharides present in the EPS antigen were mannose, galactose and glucose. The EPS obtained from cultures of M. racemosus and R. oligosporus also contained rhamnose. In the EPS produced by Penicillium spp. and A. repens the galactose residues were determined to be immunodominant.

Mucor irregularis infection and lethal midline granuloma: a case report and review of published literature.

PubMed

Li, Dong Ming; Lun, Li De

2012-12-01

Mucor irregularis (Rhizomucor variabilis) infection and lethal midline granuloma (LMG) are characterized by progressive swelling, ulceration, and destruction of the central face that is usually fatal. Pathological features are inflammation, necrosis, and granulation. LMG has been called by various names, and in recent years, it has been known as NK/T cell lymphoma. However, diagnosis still relies on the progressive necrosis course rather than malignancy in histology. The disease has long challenged physicians, particularly when it worsens with radiotherapy or chemotherapy but sometimes achieves total remission without anti-malignancy therapies. We describe a 35-year-old man who had typical clinical-pathological symptoms of LMG, which turned out to be primary M. irregularis infection; that was diagnosed by positive tissue culture and fungal elements in histology. The patient was successfully treated with antifungal therapy (liposomal amphotericin B, total 4,600 mg and amphotericin B total 277 mg, over a duration of 70 days). We hereby review current knowledge about the epidemiology, clinical manifestations, radiographic characteristics, and pathologic features of LMG with those of M. irregularis infection and their associations. We conclude that primary M. irregulars infection can mimic the clinico-pathological symptoms of LMG and the condition responds favorably to aggressive antifungal therapy.

Mucor circinelloides induces platelet aggregation through integrin αIIbβ3 and FcγRIIA.

PubMed

Ghuman, Harlene; Shepherd-Roberts, Alicia; Watson, Stephanie; Zuidscherwoude, Malou; Watson, Steve P; Voelz, Kerstin

2018-01-03

Thrombosis is a hallmark of the fatal fungal infection mucormycosis. Yet, the platelet activation pathway in response to mucormycetes is unknown. In this study we determined the platelet aggregation potential of Mucor circinelloides (M. circinelloides) NRRL3631, characterized the signaling pathway facilitating aggregation in response to fungal spores, and identified the influence of the spore developmental stage upon platelet aggregation potential. Using impedance and light-transmission aggregometry, we showed that M. circinelloides induced platelet aggregation in whole blood and in platelet-rich plasma, respectively. The formation of large spore-platelet aggregates was confirmed by light-sheet microscopy, which showed spores dispersed throughout the aggregate. Aggregation potential was dependent on the spore's developmental stage, with the strongest platelet aggregation by spores in mid-germination. Inhibitor studies revealed platelet aggregation was mediated by the low affinity IgG receptor FcγRIIA and integrin αIIbβ3; Src and Syk tyrosine kinase signaling; and the secondary mediators TxA 2 and ADP. Flow cytometry of antibody stained platelets showed that interaction with spores increased expression of platelet surface integrin αIIbβ3 and the platelet activation marker CD62P. Together, this is the first elucidation of the signaling pathways underlying thrombosis formation during a fungal infection, highlighting targets for therapeutic intervention.

Effects of Plant Growth Hormones on Mucor indicus Growth and Chitosan and Ethanol Production.

PubMed

Safaei, Zahra; Karimi, Keikhosro; Golkar, Poorandokht; Zamani, Akram

2015-07-22

The objective of this study was to investigate the effects of indole-3-acetic acid (IAA) and kinetin (KIN) on Mucor indicus growth, cell wall composition, and ethanol production. A semi-synthetic medium, supplemented with 0-5 mg/L hormones, was used for the cultivations (at 32 °C for 48 h). By addition of 1 mg/L of each hormone, the biomass and ethanol yields were increased and decreased, respectively. At higher levels, however, an inverse trend was observed. The glucosamine fraction of the cell wall, as a representative for chitosan, followed similar but sharper changes, compared to the biomass. The highest level was 221% higher than that obtained without hormones. The sum of glucosamine and N-acetyl glucosamine (chitin and chitosan) was noticeably enhanced in the presence of the hormones. Increase of chitosan was accompanied by a decrease in the phosphate content, with the lowest phosphate (0.01 g/g cell wall) being obtained when the chitosan was at the maximum (0.45 g/g cell wall). In conclusion, IAA and KIN significantly enhanced the M. indicus growth and chitosan production, while at the same time decreasing the ethanol yield to some extent. This study shows that plant growth hormones have a high potential for the improvement of fungal chitosan production by M. indicus.

Biosynthesis, characterization and enzymatic transesterification of single cell oil of Mucor circinelloides--a sustainable pathway for biofuel production.

PubMed

Carvalho, Ana K F; Rivaldi, Juan D; Barbosa, Jayne C; de Castro, Heizir F

2015-04-01

The filamentous fungus Mucor circinelloides URM 4182 was tested to determine its ability to produce single-cell oil suitable for obtaining biodiesel. Cell growth and lipid accumulation were investigated in a medium containing glucose as the main carbon source. A microwave-assisted ethanol extraction technique (microwave power ⩽200 W, 50-60 °C) was established and applied to lipid extraction from the fungal hyphae to obtain high lipid concentration (44%wt) of the dry biomass, which was considerably higher than the quantity obtained by classical solvent methods. The lipid profile showed a considerable amount of oleic acid (39.3%wt), palmitic acid (22.2%wt) and γ-linoleic acid (10.8%wt). Biodiesel was produced by transesterification of the single-cell oil with ethanol using a immobilized lipase from Candida antarctica (Novozym® 435) as the catalyst. (1)H NMR and HPLC analyses confirmed conversion of 93% of the single-cell oil from M. circinelloides into ethyl esters (FAEE). Copyright © 2015 Elsevier Ltd. All rights reserved.

Changing Epidemiology of Mucoralean Fungi: Chronic Cutaneous Infection Caused by Mucor irregularis.

PubMed

Chander, Jagdish; Kaur, Mandeep; Bhalla, Mala; Punia, Rajpal Singh; Singla, Nidhi; Bhola, Kalyani; Alastruey-Izquierdo, Ana; Stchigel, Alberto M; Guarro, Josep

2015-10-01

The fungi pertaining to order Mucorales usually cause an acute form of clinical disease called mucormycosis. A primary chronic presentation in an immunocompetent patient is a rare form of mucormycosis. Mucor irregularis is known for causing chronic cutaneous infections geographically confined to Asia, mainly in China. We describe a case of primary chronic cutaneous mucormycosis caused by M. irregularis from a new geographical niche in India, highlighting changing aspects of its epidemiology. The patient was a farmer with a history of skin lesions over the lower limb for the past 6 years. The biopsy taken from the lesions showed pauci-septate hyphae with right-angle branching on KOH wet mount as well as special fungal stains. On fungal culture, greyish-white cottony mycelial growth of Mucormycetes was obtained. The strain was finally identified as M. irregularis on macro- and microscopic features on 2 % MEA and DNA sequencing. The antifungal susceptibility was done using EUCAST broth microdilution method and was found to be susceptible to commonly used antifungal agents. The patient was started on oral itraconazole and saturated solution of potassium iodide (SSKI). While undergoing treatment for 2 months, he was lost to follow-up, however, after a year when he recently visited the hospital; the disease got completely healed with no new crops of skin lesions. Mucoralean fungi should also be suspected in cases with chronic presentation, in immunocompetent host, as there is emergence of such fungi in new endemic areas, particularly located in Asia. The role of other antifungal agents apart from amphotericin B for the treatment of chronic mucormycosis needs to be explored.

Comparative Proteome Analysis between High Lipid-Producing Strain Mucor circinelloides WJ11 and Low Lipid-Producing Strain CBS 277.49.

PubMed

Tang, Xin; Chen, Haiqin; Gu, Zhennan; Zhang, Hao; Chen, Yong Q; Song, Yuanda; Chen, Wei

2017-06-21

Mucor circinelloides is one of few oleaginous fungi that produces a useful oil rich in γ-linolenic acid, but it usually only produces <25% total lipid. Nevertheless, we isolated a new strain WJ11 that can produce up to 36% lipid of cell dry weight. In this study, we have systematically analyzed the global changes in protein levels between the high lipid-producing strain WJ11 and the low lipid-producing strain CBS 277.49 (15%, lipid/cell dry weight) at lipid accumulation phase through comparative proteome analysis. Proteome analysis demonstrated that the branched-chain amino acid and lysine metabolism, glycolytic pathway, and pentose phosphate pathway in WJ11 were up-regulated, while the activities of tricarboxylic acid cycle and branch point enzyme for synthesis of isoprenoids were retarded compared with CBS 277.49. The coordinated regulation at proteome level indicate that more acetyl-CoA and NADPH are provided for fatty acid biosynthesis in WJ11 compared with CBS 277.49.

Static magnetic field effects on proteases with fibrinolytic activity produced by Mucor subtilissimus.

PubMed

Albuquerque, Wendell; Nascimento, Thiago; Brandão-Costa, Romero; Fernandes, Thiago; Porto, Ana

2017-02-01

The influence of a static magnetic field (SMF) on crude enzyme extracts with proteolytic activity is described and discussed. Proteolytic enzymes, which hydrolyze peptide bonds, and fibrinolytic enzymes, which dissolve fibrin clots, have industrial relevance, and applicability dependent on improvements of productivity and activity. We investigated whether a moderate SMF affects proteolysis in different in vitro tests: general proteolysis of azocasein substrate, and static and dynamic fibrinolytic processes (to compare fibrin gel configuration under exposure). Crude enzyme extracts, obtained from solid state fermentation of Mucor subtilissimus UCP (Universidade Católica de Pernambuco, Recife, Brazil) 1262, were used to carry out assays under slightly heterogeneous fields: a varied vertical SMF (for tests in Eppendorf tubes, from 0.100 to 0.170 T) and a varied horizontal SMF (for tests in Petri dishes, from 0.01 to 0.122 T), generated by two permanent magnets (NdFeB alloy). Results showed significant differences (P < 0.05) in static fibrinolysis assays after 24 h of exposure. The mean diameter of halos of fibrin degradation in the treated group increased by 21% compared to the control group; and the pixel number count of fibrin consumption (in a computational analysis of the area of each halo) enhanced by 30% with exposure. However, in dynamic fibrinolysis assays, no effects of SMF were observed. These results suggest a response of fibrin monomers to the SMF as a possible cause of the observed effects. Bioelectromagnetics. 38:109-120, 2017. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

Expression of Xanthophyllomyces dendrorhous cytochrome-P450 hydroxylase and reductase in Mucor circinelloides.

PubMed

Csernetics, Árpád; Tóth, Eszter; Farkas, Anita; Nagy, Gábor; Bencsik, Ottó; Vágvölgyi, Csaba; Papp, Tamás

2015-02-01

Carotenoids are natural pigments that act as powerful antioxidants and have various beneficial effects on human and animal health. Mucor circinelloides (Mucoromycotina) is a carotenoid producing zygomycetes fungus, which accumulates β-carotene as the main carotenoid but also able to produce the hydroxylated derivatives of β-carotene (i.e. zeaxanthin and β-cryptoxanthin) in low amount. These xanthophylls, together with the ketolated derivatives of β-carotene (such as canthaxanthin, echinenone and astaxanthin) have better antioxidant activity than β-carotene. In this study our aim was to modify and enhance the xanthophyll production of the M. circinelloides by expression of heterologous genes responsible for the astaxanthin biosynthesis. The crtS and crtR genes, encoding the cytochrome-P450 hydroxylase and reductase, respectively, of wild-type and astaxanthin overproducing mutant Xanthophyllomyces dendrorhous strains were amplified from cDNA and the nucleotide and the deduced amino acid sequences were compared to each other. Introduction of the crtS on autonomously replicating plasmid in the wild-type M. circinelloides resulted enhanced zeaxanthin and β-cryptoxanthin accumulation and the presence of canthaxanthin, echinenone and astaxanthin in low amount; the β-carotene hydroxylase and ketolase activity of the X. dendrorhous cytochrome-P450 hydroxylase in M. circinelloides was verified. Increased canthaxanthin and echinenone production was observed by expression of the gene in a canthaxanthin producing mutant M. circinelloides. Co-expression of the crtR and crtS genes led to increase in the total carotenoid and slight change in xanthophyll accumulation in comparison with transformants harbouring the single crtS gene.

Identification of a β-glucosidase from the Mucor circinelloides genome by peptide pattern recognition.

PubMed

Huang, Yuhong; Busk, Peter Kamp; Grell, Morten Nedergaard; Zhao, Hai; Lange, Lene

2014-12-01

Mucor circinelloides produces plant cell wall degrading enzymes that allow it to grow on complex polysaccharides. Although the genome of M. circinelloides has been sequenced, only few plant cell wall degrading enzymes are annotated in this species. We applied peptide pattern recognition, which is a non-alignment based method for sequence analysis to map conserved sequences in glycoside hydrolase families. The conserved sequences were used to identify similar genes in the M. circinelloides genome. We found 12 different novel genes encoding members of the GH3, GH5, GH9, GH16, GH38, GH47 and GH125 families in M. circinelloides. One of the two GH3-encoding genes was predicted to encode a β-glucosidase (EC 3.2.1.21). We expressed this gene in Pichia pastoris KM71H and found that the purified recombinant protein had relative high β-glucosidase activity (1.73U/mg) at pH5 and 50°C. The Km and Vmax with p-nitrophenyl-β-d-glucopyranoside as substrate was 0.20mM and 2.41U/mg, respectively. The enzyme was not inhibited by glucose and retained 84% activity at glucose concentrations up to 140mM. Although zygomycetes are not considered to be important degraders of lignocellulosic biomass in nature, the present finding of an active β-glucosidase in M. circinelloides demonstrates that enzymes from this group of fungi have a potential for cellulose degradation. Copyright © 2014 Elsevier Inc. All rights reserved.

Identification and characterization of D-xylulokinase from the D-xylose-fermenting fungus, Mucor circinelloides.

PubMed

Komeda, Hidenobu; Yamasaki-Yashiki, Shino; Hoshino, Kazuhiro; Asano, Yasuhisa

2014-11-01

D-Xylulokinase catalyzes the phosphorylation of D-xylulose in the final step of the pentose catabolic pathway to form d-xylulose-5-phosphate. The D-xylulokinase activity was found to be induced by both D-xylose and L-arabinose, as well as some of the other enzymes involved in the pentose catabolism, in the D-xylose-fermenting zygomycetous fungus, Mucor circinelloides NBRC 4572. The putative gene, xyl3, which may encode D-xylulokinase, was detected in the genome sequence of this strain. The amino acid sequence deduced from the gene was more similar to D-xylulokinases from an animal origin than from other fungi. The recombinant enzyme was purified from the E. coli transformant expressing xyl3 and then characterized. The ATP-dependent phosphorylative activity of the enzyme was the highest toward D-xylulose. Its kinetic parameters were determined as Km (D-xylulose) = 0.29 mM and Km (ATP) = 0.51 mM, indicating that the xyl3 gene encoded D-xylulokinase (McXK). Western blot analysis revealed that McXK was induced by L-arabinose as well as D-xylose and the induction was repressed in the presence of D-glucose, suggesting that the enzyme may be involved in the catabolism of D-xylose and L-arabinose and is subject to carbon catabolite repression in this fungus. This is the first study on D-xylulokinase from zygomycetous fungi. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Distinct RNAi Pathways in the Regulation of Physiology and Development in the Fungus Mucor circinelloides.

PubMed

Ruiz-Vázquez, Rosa M; Nicolás, Francisco E; Torres-Martínez, Santiago; Garre, Victoriano

2015-01-01

The basal fungus Mucor circinelloides has become, in recent years, a valuable model to study RNA-mediated gene silencing or RNA interference (RNAi). Serendipitously discovered in the late 1900s, the gene silencing in M. circinelloides is a landscape of consensus and dissents. Although similar to other classical fungal models in the basic design of the essential machinery that is responsible for silencing of gene expression, the existence of small RNA molecules of different sizes generated during this process and the presence of a mechanism that amplifies the silencing signal, give it a unique identity. In addition, M. circinelloides combines the components of RNAi machinery to carry out functions that not only limit themselves to the defense against foreign genetic material, but it uses some of these elements to regulate the expression of its own genes. Thus, different combinations of RNAi elements produce distinct classes of endogenous small RNAs (esRNAs) that regulate different physiological and developmental processes in response to environmental signals. The recent discovery of a new RNAi pathway involved in the specific degradation of endogenous mRNAs, using a novel RNase protein, adds one more element to the exciting puzzle of the gene silencing in M. circinelloides, in addition to providing hints about the evolutionary origin of the RNAi mechanism. Copyright © 2015 Elsevier Inc. All rights reserved.

Overexpression of malic enzyme (ME) of Mucor circinelloides improved lipid accumulation in engineered Rhodotorula glutinis.

PubMed

Li, Zhi; Sun, Hanxiao; Mo, Xuemei; Li, Xiuying; Xu, Bo; Tian, Peng

2013-06-01

The oleaginous yeast Rhodotorula glutinis has been known to be a potential feedstock for lipid production. In the present study, we investigated the enhancement of expression of malic enzyme (ME; NADP(+) dependent; EC 1.1.1.40) from Mucor circinelloides as a strategy to improve lipid content inside the yeast cells. The 26S rDNA and 5.8S rDNA gene fragments isolated from Rhodotorula glutinis were used for homologous integration of ME gene into R. glutinis chromosome under the control of the constitutively highly expressed gene phosphoglycerate kinase 1 to achieve stable expression. We demonstrated that by increasing the expression of the foreign ME gene in R. glutinis, we successfully improved the lipid content by more than twofold. At the end of lipid accumulation phrase (96 h) in the transformants, activity of ME was increased by twofold and lipid content of the yeast cells was increased from 18.74 % of the biomass to 39.35 %. Simultaneously, there were no significant differences in fatty acid profiles between the wild-type strain and the recombinant strain. Over 94 % of total fatty acids were C16:0, C18:0, C16:1, C18:1, and C18:2. Our results indicated that heterologous expression of NADP(+)-dependent ME involved in fatty acid biosynthesis indeed increased the lipid accumulation in the oleaginous yeast R. glutinis.

Selection of reference genes for quantitative real time RT-PCR during dimorphism in the zygomycete Mucor circinelloides.

PubMed

Valle-Maldonado, Marco I; Jácome-Galarza, Irvin E; Gutiérrez-Corona, Félix; Ramírez-Díaz, Martha I; Campos-García, Jesús; Meza-Carmen, Víctor

2015-03-01

Mucor circinelloides is a dimorphic fungal model for studying several biological processes including cell differentiation (yeast-mold transitions) as well as biodiesel and carotene production. The recent release of the first draft sequence of the M. circinelloides genome, combined with the availability of analytical methods to determine patterns of gene expression, such as quantitative Reverse transcription-Polymerase chain reaction (qRT-PCR), and the development of molecular genetic tools for the manipulation of the fungus, may help identify M. circinelloides gene products and analyze their relevance in different biological processes. However, no information is available on M. circinelloides genes of stable expression that could serve as internal references in qRT-PCR analyses. One approach to solve this problem consists in the use of housekeeping genes as internal references. However, validation of the usability of these reference genes is a fundamental step prior to initiating qRT-PCR assays. This work evaluates expression of several constitutive genes by qRT-PCR throughout the morphological differentiation stages of M. circinelloides; our results indicate that tfc-1 and ef-1 are the most stable genes for qRT-PCR assays during differentiation studies and they are proposed as reference genes to carry out gene expression studies in this fungus.

Impact of Phosphate, Potassium, Yeast Extract, and Trace Metals on Chitosan and Metabolite Production by Mucor indicus.

PubMed

Safaei, Zahra; Karimi, Keikhosro; Zamani, Akram

2016-08-30

In this study the effects of phosphate, potassium, yeast extract, and trace metals on the growth of Mucor indicus and chitosan, chitin, and metabolite production by the fungus were investigated. Maximum yield of chitosan (0.32 g/g cell wall) was obtained in a phosphate-free medium. Reversely, cell growth and ethanol formation by the fungus were positively affected in the presence of phosphate. In a phosphate-free medium, the highest chitosan content (0.42 g/g cell wall) and cell growth (0.66 g/g sugar) were obtained at 2.5 g/L of KOH. Potassium concentration had no significant effect on ethanol and glycerol yields. The presence of trace metals significantly increased the chitosan yield at an optimal phosphate and potassium concentration (0.50 g/g cell wall). By contrast, production of ethanol by the fungus was negatively affected (0.33 g/g sugars). A remarkable increase in chitin and decrease in chitosan were observed in the absence of yeast extract and concentrations lower than 2 g/L. The maximum chitosan yield of 51% cell wall was obtained at 5 g/L of yeast extract when the medium contained no phosphate, 2.5 g/L KOH, and 1 mL/L trace metal solution.

Composition and morphology characterization of exopolymeric substances produced by the PAH-degrading fungus of Mucor mucedo.

PubMed

Jia, Chunyun; Li, Xiaojun; Allinson, Graeme; Liu, Changfeng; Gong, Zongqiang

2016-05-01

To explore the role of exopolymeric substances (EPS) in the process of polycyclic aromatic hydrocarbons (PAH) biodegradation, the characteristics of EPS isolated from a PAH-degrading fungus were investigated firstly by spectrometric determination, microscopic observation, Fourier transform-infrared spectroscopy (FT-IR), and three-dimensional excitation-emission matrix fluorescence spectroscopy (3D-EEM), and then the PAH-degrading ability of isolated EPS was evaluated. The EPS compositions and morphology varied significantly with the extraction methods. EPS were mainly composed of proteins, carbohydrate, and humic-like substances, and the cation exchange resin (CER)-extracted EPS were granular while other EPS samples were all powders. Heating was the most effective treatment method, followed by the CER, centrifugation, and ultrasonication methods. However, 3D-EEM data demonstrated that heating treatment makes the mycelia lyse the most. Overall, therefore, the CER was the best EPS extraction method for Mucor mucedo (M. mucedo). The PAH degradation results indicated that 87 % of pyrene and 81 % of benzo[a]pyrene (B[a]P) were removed by M. mucedo over 12 days and 9 % more pyrene and 7 % more B[a]P were reduced after CER-extracted EPS addition of 465 mg l(-1). The investigation of EPS characterization and EPS enhancing PAH biodegradation is the premise for further in-depth exploration of the role of EPS contribution to PAH biodegradation.

Two new β-glucosidases from ethanol-fermenting fungus Mucor circinelloides NBRC 4572: enzyme purification, functional characterization, and molecular cloning of the gene.

PubMed

Kato, Yasuo; Nomura, Taiji; Ogita, Shinjiro; Takano, Maki; Hoshino, Kazuhiro

2013-12-01

Two β-glucosidases (BGLs 1 and 2) were purified to homogeneity from the extracellular enzyme preparations of the ethanol-fermenting Mucor circinelloides NBRC 4572 statically grown on rice straw. BGLs 1 and 2 are monomeric glycoproteins whose apparent molecular masses (Ms) are around 78 kDa, which decreased by approximately 10 kDa upon enzymatic deglycosylation. Both BGLs showed similar enzyme characteristics in optimal temperature and pH, stability, and inhibitors. They were active against a wide range of aryl-β-glucosides and β-linked glucose oligosaccharides. Their amino acid sequences shared 81% identity and exhibited less than 60% identity with the known family-3 BGLs. Considering properties such as reduced inhibition by ethanol, glucose, and cellobiose, low transglucosylation activity, wider substrate range, less binding affinity to lignocellulosic materials, and abundant expression, BGL1 is likely to be more suitable for bioethanol production than BGL2 via simultaneous saccharification and fermentation of rice straw with M. circinelloides.

Effects of 20 standard amino acids on the growth, total fatty acids production, and γ-linolenic acid yield in Mucor circinelloides.

PubMed

Tang, Xin; Zhang, Huaiyuan; Chen, Haiqin; Chen, Yong Q; Chen, Wei; Song, Yuanda

2014-12-01

Twenty standard amino acids were examined as single nitrogen source on the growth, total fatty acids production, and yield of γ-linolenic acid (GLA) in Mucor circinelloides. Of the amino acids, tyrosine gave the highest biomass and lipid accumulation and thus resulted in a high GLA yield with respective values of 17.8 g/L, 23 % (w/w, dry cell weight, DCW), and 0.81 g/L, which were 36, 25, and 72 % higher than when the fungus was grown with ammonium tartrate. To find out the potential mechanism underlying the increased lipid accumulation of M. circinelloides when grown on tyrosine, the activity of lipogenic enzymes of the fungus during lipid accumulation phase was measured. The enzyme activities of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase, and ATP-citrate lyase were up-regulated, while NADP-isocitrate dehydrogenase was down-regulated by tyrosine during the lipid accumulation phase of the fungus which suggested that these enzymes may be involved in the increased lipid biosynthesis by tyrosine in this fungus.

Robust Phagocyte Recruitment Controls the Opportunistic Fungal Pathogen Mucor circinelloides in Innate Granulomas In Vivo

PubMed Central

2018-01-01

ABSTRACT Mucormycosis is an emerging fungal infection with extremely high mortality rates in patients with defects in their innate immune response, specifically in functions mediated through phagocytes. However, we currently have a limited understanding of the molecular and cellular interactions between these innate immune effectors and mucormycete spores during the early immune response. Here, the early events of innate immune recruitment in response to infection by Mucor circinelloides spores are modeled by a combined in silico modeling approach and real-time in vivo microscopy. Phagocytes are rapidly recruited to the site of infection in a zebrafish larval model of mucormycosis. This robust early recruitment protects from disease onset in vivo. In silico analysis identified that protection is dependent on the number of phagocytes at the infection site, but not the speed of recruitment. The mathematical model highlights the role of proinflammatory signals for phagocyte recruitment and the importance of inhibition of spore germination for protection from active fungal disease. These in silico data are supported by an in vivo lack of fungal spore killing and lack of reactive oxygen burst, which together result in latent fungal infection. During this latent stage of infection, spores are controlled in innate granulomas in vivo. Disease can be reactivated by immunosuppression. Together, these data represent the first in vivo real-time analysis of innate granuloma formation during the early stages of a fungal infection. The results highlight a potential latent stage during mucormycosis that should urgently be considered for clinical management of patients. PMID:29588406

[Blue-light induced expression of S-adenosy-L-homocysteine hydrolase-like gene in Mucor amphibiorum RCS1].

PubMed

Gao, Ya; Wang, Shu; Fu, Mingjia; Zhong, Guolin

2013-09-04

To determine blue-light induced expression of S-adenosyl-L-homocysteine hydrolase-like (sahhl) gene in fungus Mucor amphibiorum RCS1. In the random process of PCR, a sequence of 555 bp was obtained from M. amphibiorum RCS1. The 555 bp sequence was labeled with digoxin to prepare the probe for northern hybridization. By northern hybridization, the transcription of sahhl gene was analyzed in M. amphibiorum RCS1 mycelia culture process from darkness to blue light to darkness. Simultaneously real-time PCR method was used to the sahhl gene expression analysis. Compared with the sequence of sahh gene from Homo sapiens, Mus musculus and some fungi species, a high homology of the 555 bp sequence was confirmed. Therefore, the preliminary confirmation has supported that the 555 bp sequence should be sahhl gene from M. amphibiorum RCS1. Under the dark pre-culture in 24 h, a large amounts of transcript of sahhl gene in the mycelia can be detected by northern hybridization and real-time PCR in the condition of 24 h blue light. But a large amounts of transcript of sahhl gene were not found in other detection for the dark pre-culture of 48 h, even though M. amphibiorum RCS1 mycelia were induced by blue light. Blue light can induce the expression of sahhl gene in the vigorous growth of M. amphibiorum RCS1 mycelia.

Biosurfactant production by Mucor circinelloides on waste frying oil and possible uses in crude oil remediation.

PubMed

Hasanizadeh, Parvin; Moghimi, Hamid; Hamedi, Javad

2017-10-01

Biosurfactants are biocompatible surface active agents which many microorganisms produce. This study investigated the production of biosurfactants by Mucor circinelloides. The effects of different factors on biosurfactant production, including carbon sources and concentrations, nitrogen sources, and iron (II) concentration, were studied and the optimum condition determined. Finally, the strain's ability to remove the crude oil and its relationship with biosurfactant production was evaluated. The results showed that M. circinelloides could reduce the surface tension of the culture medium to 26.6 mN/m and create a clear zone of 12.9 cm diameter in an oil-spreading test. The maximum surface tension reduction was recorded 3 days after incubation. The optimum condition for biosurfactant production was achieved in the presence of 8% waste frying oil as a carbon source, 2 g/L yeast extract as a nitrogen source, and 0.01 mM FeSO 4 . M. circinelloides could consume 8% waste frying oil in 5 days of incubation, and 87.6% crude oil in 12 days of incubation. A direct correlation was observed between oil degradation and surface tension reduction in the first 3 days of fungal growth. The results showed that the waste frying oil could be recommended as an inexpensive oily waste substance for biosurfactant production, and M. circinelloides could have the potential to treat waste frying oil. According to the results, the produced crude biosurfactant or fungal strain could be directly used for the mycoremediation of crude oil contamination in oil fields.

Fungal strains isolated from cork stoppers and the formation of 2,4,6-trichloroanisole involved in the cork taint of wine.

PubMed

Prak, Sina; Gunata, Ziya; Guiraud, Joseph-Pierre; Schorr-Galindo, Sabine

2007-05-01

Cork taint is mainly due to 2,4,6-trichloroanisole (TCA) produced through the activity of undesirable fungal strains. We observed that CFU mould number in TCA-containing stoppers was not quantitatively different to that of the stoppers not containing TCA (ca. 10(5)CFU/g). In contrast more fungi diversity was observed in TCA-containing stoppers. Penicillium spp (Penicillium chrysogenum, Penicillium glabrum), Aspergillus spp (Aspergillus niger and Aspergillus oryzae), Chrysonilia sitophila, Mucor racemosus, Paecilomyces sp. and Trichoderma viride were found in TCA-containing stoppers, while C. sitophila and Penicillium sp. were the main fungi in the stoppers devoid of TCA. Conidia were numerous close to the lenticels and present from the lateral surface through to the centre of the stoppers. Strains of Aspergillus, Mucor, Paecilomyces, Penicillium and Trichoderma isolated from TCA-containing stoppers were able to convert 2,4,6-trichlorophenol (TCP) in TCA in resting cell or growing conditions. The best yields of conversion were obtained by green fungi Paecilomyces sp. and P. chrysogenum, 17% and 20%, respectively. Chysonilia sitophila and Penicillium sp. did not produce TCA from TCP in our conditions.

Generation of a Mutant Mucor hiemalis Endoglycosidase That Acts on Core-fucosylated N-Glycans.

PubMed

Katoh, Toshihiko; Katayama, Takane; Tomabechi, Yusuke; Nishikawa, Yoshihide; Kumada, Jyunichi; Matsuzaki, Yuji; Yamamoto, Kenji

2016-10-28

Endo-β-N-acetylglucosaminidase M (Endo-M), an endoglycosidase from the fungus Mucor hiemalis, is a useful tool for chemoenzymatic synthesis of glycoconjugates, including glycoprotein-based therapeutics having a precisely defined glycoform, by virtue of its transglycosylation activity. Although Endo-M has been known to act on various N-glycans, it does not act on core-fucosylated N-glycans, which exist widely in mammalian glycoproteins, thus limiting its application. Therefore, we performed site-directed mutagenesis on Endo-M to isolate mutant enzymes that are able to act on mammalian-type core-α1,6-fucosylated glycans. Among the Endo-M mutant enzymes generated, those in which the tryptophan at position 251 was substituted with alanine or asparagine showed altered substrate specificities. Such mutant enzymes exhibited increased hydrolysis of a synthetic α1,6-fucosylated trimannosyl core structure, whereas their activity on the afucosylated form decreased. In addition, among the Trp-251 mutants, the W251N mutant was most efficient in hydrolyzing the core-fucosylated substrate. W251N mutants could act on the immunoglobulin G-derived core-fucosylated glycopeptides and human lactoferrin glycoproteins. This mutant was also capable of transferring the sialyl glycan from an activated substrate intermediate (sialyl glyco-oxazoline) onto an α1,6-fucosyl-N-acetylglucosaminyl biotin. Furthermore, the W251N mutant gained a glycosynthase-like activity when a N175Q substitution was introduced and it caused accumulation of the transglycosylation products. These findings not only give insights into the substrate recognition mechanism of glycoside hydrolase family 85 enzymes but also widen their scope of application in preparing homogeneous glycoforms of core-fucosylated glycoproteins for the production of potent glycoprotein-based therapeutics. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Apoptosis-like death was involved in freeze-drying-preserved fungus Mucor rouxii and can be inhibited by L-proline.

PubMed

Wang, Xiaoyun; Wang, Youzhi

2016-02-01

Freeze-drying is one of the most effective methods to preserve fungi for an extended period. However, it is associated with a loss of viability and shortened storage time in some fungi. This study evaluated the stresses that led to the death of freeze-dried Mucor rouxii by using cell apoptotic methods. The results showed there were apoptosis-inducing stresses, such as the generation of obvious intracellular reactive oxygen species (ROS) and metacaspase activation. Moreover, nuclear condensation and a delayed cell death peak were determined after rehydration and 24 h incubation in freeze-dried M. rouxii via a propidium iodide (PI) assay, which is similar to the phenomenon of cryopreservation-induced delayed-onset cell death (CIDOCD). Then, several protective agents were tested to decrease the apoptosis-inducing stresses and to improve the viability. Finally, it was found that 1.6 mM L-proline can effectively decrease the nuclear condensation rate and increase the survival rate in freeze-dried M. rouxii. (1) apoptosis-inducing factors occur in freeze-dried M. rouxii. (2) ROS and activated metacaspases lead to death in freeze-dried M. rouxii. (3)L-proline increases the survival rate of freeze-dried M. rouxii. Copyright © 2015 Elsevier Inc. All rights reserved.

A thermolabile aspartic proteinase from Mucor mucedo DSM 809: gene identification, cloning, and functional expression in Pichia pastoris.

PubMed

Yegin, Sirma; Fernandez-Lahore, Marcelo

2013-06-01

In this study, the cDNA encoding the aspartic proteinase of Mucor mucedo DSM 809 has been identified by RNA ligased-mediated and oligo-capping rapid amplification of cDNA ends (RACE) technique. The gene contained an open reading frame of 1,200 bp and encoded for a signal peptide of 21 amino acid residues. Two N-glycosylation sites were observed within the identified sequence. The proteinase gene was cloned into the vector pGAPZαA and expressed in Pichia pastoris X-33 for the first time. The protein has been secreted in functionally active form into the culture medium. The expression system does not require any acid activation process. The factors affecting the expression level were optimized in shaking flask cultures. Maximum enzyme production was observed with an initial medium pH of 3.5 at 20 °C and 220 rpm shaking speed utilizing 4 % glucose as a carbon and energy source. The enzyme was purified with cation exchange chromatography and further studies revealed that the enzyme was secreted in glycosylated form. The purified enzyme exhibited remarkable sensitivity to thermal treatment and became completely inactivated after incubation at 55 °C for 10 min. These results indicated that the recombinant proteinase could be considered as a potential rennet candidate for the cheese-making industry.

Improved γ-linolenic acid production in Mucor circinelloides by homologous overexpressing of delta-12 and delta-6 desaturases.

PubMed

Zhang, Yao; Luan, Xiao; Zhang, Huaiyuan; Garre, Victoriano; Song, Yuanda; Ratledge, Colin

2017-06-21

γ-Linolenic acid (GLA) is important because of its nutritional value and medicinal applications. Although the biosynthetic pathways of some plant and microbial GLA have been deciphered, current understanding of the correlation between desaturases and GLA synthesis in oleaginous fungi is incomplete. In previous work, we found that a large amount of oleic acid (OA) had not been converted to linoleic acid (LA) or GLA in Mucor circinelloides CBS 277.49, which may be due to inadequate activities of the delta-12 or delta-6 desaturases, and thus leading to the accumulation of OA and LA. Thus, it is necessary to explore the main contributing factor during the process of GLA biosynthesis in M. circinelloides. To enhance GLA production in M. circinelloides, homologous overexpression of delta-12 and two delta-6 desaturases (named delta-6-1 and delta-6-2, respectively) were analyzed. When delta-6 desaturase were overexpressed in M. circinelloides, up to 43% GLA was produced in the total fatty acids, and the yield of GLA reached 180 mg/l, which were, respectively, 38 and 33% higher than the control strain. These findings revealed that delta-6 desaturase (especially for delta-6-1 desaturase) plays an important role in GLA synthesis by M. circinelloides. The strain overexpressing delta-6-1 desaturase may have potential application in microbial GLA production.

Biochemical mechanism of phytoremediation process of lead and cadmium pollution with Mucor circinelloides and Trichoderma asperellum.

PubMed

Zhang, Xu; Li, Xinxin; Yang, Huanhuan; Cui, Zhaojie

2018-08-15

This study focused on the bioremediation mechanisms of lead (0, 100, 500, 1000 mg kg -1 ) and cadmium (0,10,50,100 mg kg -1 ) contaminated soil using two indigenous fungi selected from mine tailings as the phytostimulation of Arabidopsis thaliana. The two fungal strains were characterized as Mucor circinelloides (MC) and Trichoderma asperellum (TA) by internal transcribed spacer sequencing at the genetic levels. Our research revealed that Cadmium was more toxic to plant growth than lead and meanwhile, MC and TA can strengthen A. thaliana tolerance to cadmium and lead with 40.19-117.50% higher root length and 58.31-154.14% shoot fresh weight of plant compared to non-inoculation. In this study, TA exhibited a higher potential to the inactivation of cadmium; however, MC was more effective in lead passivation. There was a direct correlation between the type of fungi, heavy metal content, heavy metal type and oxidative damage in plant. Both lead and cadmium induced oxidative damage as indicated by increased superoxide dismutase and catalase activities, while the antioxidant levels were significantly higher in fungal inoculated plants compared with those non-inoculated. The analysis of soil enzyme activity and taxonomic richness uncovered that the dominant structures of soil microbial community were altered by exogenous microbial agents. MC enhanced higher microbial diversity and soil enzyme activity than TA. The two indigenous fungi lessened several limiting factors with respect to phytoremediation technology, such as soil chemistry, contamination level and transformation, and metal solubility. Copyright © 2018 Elsevier Inc. All rights reserved.

Phylogenetic analysis of fungal heterotrimeric G protein-encoding genes and their expression during dimorphism in Mucor circinelloides.

PubMed

Valle-Maldonado, Marco Iván; Jácome-Galarza, Irvin Eduardo; Díaz-Pérez, Alma Laura; Martínez-Cadena, Guadalupe; Campos-García, Jesús; Ramírez-Díaz, Martha Isela; Reyes-De la Cruz, Homero; Riveros-Rosas, Héctor; Díaz-Pérez, César; Meza-Carmen, Víctor

2015-12-01

In fungi, heterotrimeric G proteins are key regulators of biological processes such as mating, virulence, morphology, among others. Mucor circinelloides is a model organism for many biological processes, and its genome contains the largest known repertoire of genes that encode putative heterotrimeric G protein subunits in the fungal kingdom: twelve Gα (McGpa1-12), three Gβ (McGpb1-3), and three Gγ (McGpg1-3). Phylogenetic analysis of fungal Gα showed that they are divided into four distinct groups as reported previously. Fungal Gβ and Gγ are also divided into four phylogenetic groups, and to our understanding this is the first report of a phylogenetic classification for fungal Gβ and Gγ subunits. Almost all genes that encode putative heterotrimeric G subunits in M. circinelloides are differentially expressed during dimorphic growth, except for McGpg1 (Gγ) that showed very low mRNA levels at all developmental stages. Moreover, several of the subunits are expressed in a similar pattern and at the same level, suggesting that they constitute discrete complexes. For example, McGpb3 (Gβ), and McGpg2 (Gγ), are co-expressed during mycelium growth, and McGpa1, McGpb2, and McGpg2, are co-expressed during yeast development. These findings provide the conceptual framework to study the biological role of these genes during M. circinelloides morphogenesis. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Simultaneous Detection and Identification of Aspergillus and Mucorales Species in Tissues Collected from Patients with Fungal Rhinosinusitis▿

PubMed Central

Zhao, Zuotao; Li, Lili; Wan, Zhe; Chen, Wei; Liu, Honggang; Li, Ruoyu

2011-01-01

Rapid detection and differentiation of Aspergillus and Mucorales species in fungal rhinosinusitis diagnosis are desirable, since the clinical management and prognosis associated with the two taxa are fundamentally different. We describe an assay based on a combination of broad-range PCR amplification and reverse line blot hybridization (PCR/RLB) to detect and differentiate the pathogens causing fungal rhinosinusitis, which include five Aspergillus species (A. fumigatus, A. flavus, A. niger, A. terreus, and A. nidulans) and seven Mucorales species (Mucor heimalis, Mucor racemosus, Mucor cercinelloidea, Rhizopus arrhizus, Rhizopus microsporus, Rhizomucor pusillus, and Absidia corymbifera). The assay was validated with 98 well-characterized clinical isolates and 41 clinical tissue specimens. PCR/RLB showed high sensitivity and specificity, with 100% correct identifications of 98 clinical isolates and no cross-hybridization between the species-specific probes. Results for five control isolates, Candida albicans, Fusarium solani, Scedosporium apiospermum, Penicillium marneffei, and Exophiala verrucosa, were negative as judged by PCR/RLB. The analytical sensitivity of PCR/RLB was found to be 1.8 × 10−3 ng/μl by 10-fold serial dilution of Aspergillus genomic DNA. The assay identified 35 of 41 (85.4%) clinical specimens, exhibiting a higher sensitivity than fungal culture (22 of 41; 53.7%) and direct sequencing (18 of 41; 43.9%). PCR/RLB similarly showed high specificity, with correct identification 16 of 18 specimens detected by internal transcribed spacer (ITS) sequencing and 16 of 22 detected by fungal culture, but it also has the additional advantage of being able to detect mixed infection in a single clinical specimen. The PCR/RLB assay thus provides a rapid and reliable option for laboratory diagnosis of fungal rhinosinusitis. PMID:21325541

Simultaneous detection and identification of Aspergillus and mucorales species in tissues collected from patients with fungal rhinosinusitis.

PubMed

Zhao, Zuotao; Li, Lili; Wan, Zhe; Chen, Wei; Liu, Honggang; Li, Ruoyu

2011-04-01

Rapid detection and differentiation of Aspergillus and Mucorales species in fungal rhinosinusitis diagnosis are desirable, since the clinical management and prognosis associated with the two taxa are fundamentally different. We describe an assay based on a combination of broad-range PCR amplification and reverse line blot hybridization (PCR/RLB) to detect and differentiate the pathogens causing fungal rhinosinusitis, which include five Aspergillus species (A. fumigatus, A. flavus, A. niger, A. terreus, and A. nidulans) and seven Mucorales species (Mucor heimalis, Mucor racemosus, Mucor cercinelloidea, Rhizopus arrhizus, Rhizopus microsporus, Rhizomucor pusillus, and Absidia corymbifera). The assay was validated with 98 well-characterized clinical isolates and 41 clinical tissue specimens. PCR/RLB showed high sensitivity and specificity, with 100% correct identifications of 98 clinical isolates and no cross-hybridization between the species-specific probes. Results for five control isolates, Candida albicans, Fusarium solani, Scedosporium apiospermum, Penicillium marneffei, and Exophiala verrucosa, were negative as judged by PCR/RLB. The analytical sensitivity of PCR/RLB was found to be 1.8 × 10(-3) ng/μl by 10-fold serial dilution of Aspergillus genomic DNA. The assay identified 35 of 41 (85.4%) clinical specimens, exhibiting a higher sensitivity than fungal culture (22 of 41; 53.7%) and direct sequencing (18 of 41; 43.9%). PCR/RLB similarly showed high specificity, with correct identification 16 of 18 specimens detected by internal transcribed spacer (ITS) sequencing and 16 of 22 detected by fungal culture, but it also has the additional advantage of being able to detect mixed infection in a single clinical specimen. The PCR/RLB assay thus provides a rapid and reliable option for laboratory diagnosis of fungal rhinosinusitis.

Uptake and biotransformation of pure commercial microcystin-LR versus microcystin-LR from a natural cyanobacterial bloom extract in the aquatic fungus Mucor hiemalis.

PubMed

Esterhuizen-Londt, Maranda; Hertel, Stefanie; Pflugmacher, Stephan

2017-10-01

To evaluate the remediation efficiency of Mucor hiemalis by comparing media elimination, uptake, and biotransformation of microcystin-LR with exposure to pure toxin versus a crude bloom extract. With exposure to the extract, the elimination rate of microcystin-LR from the media, which was 0.28 ng MC-LR l -1  h -1 , was significantly higher compared to that achieved with exposure to the pure toxin (0.16 ng MC-LR l -1  h -1 ) after 24 h. However, intracellular breakdown of microcystin-LR was significantly lower in the extract exposed pellets compared to the pure toxin treated fungal pellets over time. This coincided with reduced intracellular glutathione S-transferase activity with crude extract exposure which could be responsible for the detection of only the glutathione conjugate of microcystin-LR. This paper signifies the importance of using laboratory exposure scenarios which resemble conditions in nature to fully understand and evaluate remediation efficiency. There is merit in using M. hiemalis for mycoremediation of cyanotoxins in surface waters.

A new mercury-accumulating Mucor hiemalis strain EH8 from cold sulfidic spring water biofilms.

PubMed

Hoque, Enamul; Fritscher, Johannes

2016-10-01

Here, we report about a unique aquatic fungus Mucor hiemalisEH8 that can remove toxic ionic mercury from water by intracellular accumulation and reduction into elemental mercury (Hg 0 ). EH8 was isolated from a microbial biofilm grown in sulfidic-reducing spring water sourced at a Marching's site located downhill from hop cultivation areas with a history of mercury use. A thorough biodiversity survey and mercury-removal function analyses were undertaken in an area of about 200 km 2 in Bavaria (Germany) to find the key biofilm and microbe for mercury removal. After a systematic search using metal removal assays we identified Marching spring's biofilm out of 18 different sulfidic springs' biofilms as the only one that was capable of removing ionic Hg from water. EH8 was selected, due to its molecular biological identification as the key microorganism of this biofilm with the capability of mercury removal, and cultivated as a pure culture on solid and in liquid media to produce germinating sporangiospores. They removed 99% of mercury from water within 10-48 h after initial exposure to Hg(II). Scanning electron microscopy demonstrated occurrence of intracellular mercury in germinating sporangiospores exposed to mercury. Not only associated with intracellular components, but mercury was also found to be released and deposited as metallic-shiny nanospheres. Electron-dispersive x-ray analysis of such a nanosphere confirmed presence of mercury by the HgM α peak at 2.195 keV. Thus, a first aquatic eukaryotic microbe has been found that is able to grow even at low temperature under sulfur-reducing conditions with promising performance in mercury removal to safeguard our environment from mercury pollution. © 2016 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Rapamycin exerts antifungal activity in vitro and in vivo against Mucor circinelloides via FKBP12-dependent inhibition of Tor.

PubMed

Bastidas, Robert J; Shertz, Cecelia A; Lee, Soo Chan; Heitman, Joseph; Cardenas, Maria E

2012-03-01

The zygomycete Mucor circinelloides is an opportunistic fungal pathogen that commonly infects patients with malignancies, diabetes mellitus, and solid organ transplants. Despite the widespread use of antifungal therapy in the management of zygomycosis, the incidence of infections continues to rise among immunocompromised individuals. In this study, we established that the target and mechanism of antifungal action of the immunosuppressant rapamycin in M. circinelloides are mediated via conserved complexes with FKBP12 and a Tor homolog. We found that spontaneous mutations that disrupted conserved residues in FKBP12 conferred rapamycin and FK506 resistance. Disruption of the FKBP12-encoding gene, fkbA, also conferred rapamycin and FK506 resistance. Expression of M. circinelloides FKBP12 (McFKBP12) complemented a Saccharomyces cerevisiae mutant strain lacking FKBP12 to restore rapamycin sensitivity. Expression of the McTor FKBP12-rapamycin binding (FRB) domain conferred rapamycin resistance in S. cerevisiae, and McFKBP12 interacted in a rapamycin-dependent fashion with the McTor FRB domain in a yeast two-hybrid assay, validating McFKBP12 and McTor as conserved targets of rapamycin. We showed that in vitro, rapamycin exhibited potent growth inhibitory activity against M. circinelloides. In a Galleria mellonella model of systemic mucormycosis, rapamycin improved survival by 50%, suggesting that rapamycin and nonimmunosuppressive analogs have the potential to be developed as novel antifungal therapies for treatment of patients with mucormycosis.

Comparative Studies of Oleaginous Fungal Strains (Mucor circinelloides and Trichoderma reesei) for Effective Wastewater Treatment and Bio-Oil Production

PubMed Central

Bhanja, Anshuman; Kalyanraman, V.

2014-01-01

Biological wastewater treatment typically requires the use of bacteria for degradation of carbonaceous and nitrogenous compounds present in wastewater. The high lipid containing biomass can be used to extract oil and the contents can be termed as bio-oil (or biodiesel or myco-diesel after transesterification). The separate experiments were conducted on actual wastewater samples with 5% v/v inoculum of Mucor circinelloides MTCC1297 and Trichoderma reesei NCIM992 strains. The observed reductions in chemical oxygen demand (COD) were 88.72% and 86.75% in 96 hrs and the observed substrate based biomass yields were 0.21 mg VSS/mg COD and 0.22 mg VSS/mg COD for M. circinelloides reactor and for T. reesei reactor, respectively. The resulted bio-oil production from wastewater treatment by M. circinelloides and T. reesei reactors was 142.2 mg/L and 74.1 mg/L, whereas biomass containing bio-oil contents (%w/w) were 22.11% and 9.82%, respectively. In this experiment, the fungal wastewater treatment was also compared with conventional bacterial process with respect to specific growth rate, biomass production, and oil content. This study suggests that wastewater can be used as a potential feedstock for bio-oil production with the use of oleaginous fungal strains and which could be a possible route of waste to energy. PMID:25530884

Comparative Studies of Oleaginous Fungal Strains (Mucor circinelloides and Trichoderma reesei) for Effective Wastewater Treatment and Bio-Oil Production.

PubMed

Bhanja, Anshuman; Minde, Gauri; Magdum, Sandip; Kalyanraman, V

2014-01-01

Biological wastewater treatment typically requires the use of bacteria for degradation of carbonaceous and nitrogenous compounds present in wastewater. The high lipid containing biomass can be used to extract oil and the contents can be termed as bio-oil (or biodiesel or myco-diesel after transesterification). The separate experiments were conducted on actual wastewater samples with 5% v/v inoculum of Mucor circinelloides MTCC1297 and Trichoderma reesei NCIM992 strains. The observed reductions in chemical oxygen demand (COD) were 88.72% and 86.75% in 96 hrs and the observed substrate based biomass yields were 0.21 mg VSS/mg COD and 0.22 mg VSS/mg COD for M. circinelloides reactor and for T. reesei reactor, respectively. The resulted bio-oil production from wastewater treatment by M. circinelloides and T. reesei reactors was 142.2 mg/L and 74.1 mg/L, whereas biomass containing bio-oil contents (%w/w) were 22.11% and 9.82%, respectively. In this experiment, the fungal wastewater treatment was also compared with conventional bacterial process with respect to specific growth rate, biomass production, and oil content. This study suggests that wastewater can be used as a potential feedstock for bio-oil production with the use of oleaginous fungal strains and which could be a possible route of waste to energy.

Endogenous short RNAs generated by Dicer 2 and RNA-dependent RNA polymerase 1 regulate mRNAs in the basal fungus Mucor circinelloides

PubMed Central

Nicolas, Francisco Esteban; Moxon, Simon; de Haro, Juan P.; Calo, Silvia; Grigoriev, Igor V.; Torres-Martínez, Santiago; Moulton, Vincent; Ruiz-Vázquez, Rosa M.; Dalmay, Tamas

2010-01-01

Endogenous short RNAs (esRNAs) play diverse roles in eukaryotes and usually are produced from double-stranded RNA (dsRNA) by Dicer. esRNAs are grouped into different classes based on biogenesis and function but not all classes are present in all three eukaryotic kingdoms. The esRNA register of fungi is poorly described compared to other eukaryotes and it is not clear what esRNA classes are present in this kingdom and whether they regulate the expression of protein coding genes. However, evidence that some dicer mutant fungi display altered phenotypes suggests that esRNAs play an important role in fungi. Here, we show that the basal fungus Mucor circinelloides produces new classes of esRNAs that map to exons and regulate the expression of many protein coding genes. The largest class of these exonic-siRNAs (ex-siRNAs) are generated by RNA-dependent RNA Polymerase 1 (RdRP1) and dicer-like 2 (DCL2) and target the mRNAs of protein coding genes from which they were produced. Our results expand the range of esRNAs in eukaryotes and reveal a new role for esRNAs in fungi. PMID:20427422

Can fungi compete with marine sources for chitosan production?

PubMed

Ghormade, V; Pathan, E K; Deshpande, M V

2017-11-01

Chitosan, a β-1,4-linked glucosamine polymer is formed by deacetylation of chitin. It has a wide range of applications from agriculture to human health care products. Chitosan is commercially produced from shellfish, shrimp waste, crab and lobster processing using strong alkalis at high temperatures for long time periods. The production of chitin and chitosan from fungal sources has gained increased attention in recent years due to potential advantages in terms of homogenous polymer length, high degree of deacetylation and solubility over the current marine source. Zygomycetous fungi such as Absidia coerulea, Benjaminiella poitrasii, Cunninghamella elegans, Gongrenella butleri, Mucor rouxii, Mucor racemosus and Rhizopus oryzae have been studied extensively. Isolation of chitosan are reported from few edible basidiomycetous fungi like Agaricus bisporus, Lentinula edodes and Pleurotus sajor-caju. Other organisms from mycotech industries explored for chitosan production are Aspergillus niger, Penicillium chrysogenum, Saccharomyces cerevisiae and other wine yeasts. Number of aspects such as value addition to the existing applications of fungi, utilization of waste from agriculture sector, and issues and challenges for the production of fungal chitosan to compete with existing sources, metabolic engineering and novel applications have been discussed to adjudge the potential of fungal sources for commercial chitosan production. Copyright © 2017 Elsevier B.V. All rights reserved.

Rapid Identification of Airborne Biological Particles by Flow Cytometry, Gas Chromatography, and Genetic Probes.

DTIC Science & Technology

1997-09-01

CONCLUSIONS ...................................... 36 REFERENCES ....................................... 39 -5- FIGURES la. Cladosporium and Mucor ...15 lb. Cladosporium and Heterobasidion .................... 16 Ic. Cladosporium and Penicillium ....................... 17 id. Mucor ...34. ........................................ 27 2b. "Blank 2". ........................................ 28 2c. Mucor spores ..................................... 29 2d. Aspergillus spores

Species Identification and Delineation of Pathogenic Mucorales by Matrix-Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry.

PubMed

Shao, Jin; Wan, Zhe; Li, Ruoyu; Yu, Jin

2018-04-01

This study aimed to validate the effectiveness of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS)-based identification of filamentous fungi of the order Mucorales. A total of 111 isolates covering six genera preserved at the Research Center for Medical Mycology of Peking University were selected for MALDI-TOF MS analysis. We emphasized the study of 23 strains of Mucor irregularis predominantly isolated from patients in China. We first used the Bruker Filamentous Fungi library (v1.0) to identify all 111 isolates. To increase the identification rate, we created a compensatory in-house database, the Beijing Medical University (BMU) database, using 13 reference strains covering 6 species, including M. irregularis , Mucor hiemalis , Mucor racemosus , Cunninghamella bertholletiae , Cunninghamella phaeospora , and Cunninghamella echinulata All 111 isolates were then identified by MALDI-TOF MS using a combination of the Bruker library and BMU database. MALDI-TOF MS identified 55 (49.5%) and 74 (66.7%) isolates at the species and genus levels, respectively, using the Bruker Filamentous Fungi library v1.0 alone. A combination of the Bruker library and BMU database allowed MALDI-TOF MS to identify 90 (81.1%) and 111 (100%) isolates at the species and genus levels, respectively, with a significantly increased accuracy rate. MALDI-TOF MS poorly identified Mucorales when the Bruker library was used alone due to its lack of some fungal species. In contrast, this technique perfectly identified M. irregularis after main spectrum profiles (MSPs) of relevant reference strains were added to the Bruker library. With an expanded Bruker library, MALDI-TOF MS is an effective tool for the identification of pathogenic Mucorales. Copyright © 2018 American Society for Microbiology.

Effects of post-harvest treatment using chitosan from Mucor circinelloides on fungal pathogenicity and quality of table grapes during storage.

PubMed

de Oliveira, Carlos Eduardo Vasconcelos; Magnani, Marciane; de Sales, Camila Veríssimo; Pontes, Alline Lima de Souza; Campos-Takaki, Galba Maria; Stamford, Thayza Christina Montenegro; de Souza, Evandro Leite

2014-12-01

The aim of this study was to extract chitosan (CHI) from Mucor circinelloides UCP 050 grown in a corn steep liquor (CSL)-based medium under optimized conditions and to assess the efficacy of the obtained CHI to inhibit the post-harvest pathogenic fungi Aspergillus niger URM 5162 and Rhizopus stolonifer URM 3482 in laboratory media and as a coating on table grapes (Vitis labrusca L.). The effect of CHI coating on some physical, physicochemical and sensory characteristics of the fruits during storage was assessed. The greatest amount of CHI was extracted from M. circinelloides UCP 050 grown in medium containing 7 g of CSL per 100 mL at pH 5.5 with rotation at 180 rpm. CHI from M. circinelloides UCP 050 caused morphological changes in the spores of the fungal strains tested and inhibited mycelial growth and spore germination. CHI coating delayed the growth of the assayed fungal strains in artificially infected grapes, as well as autochthonous mycoflora during storage. CHI coating preserved the quality of grapes during storage, as measured by their physical, physicochemical and sensory attributes. These results demonstrate that edible coatings derived from M. circinelloides CHI could be a useful alternative for controlling pathogenic fungi and maintaining the post-harvest quality of table grapes. Copyright © 2014 Elsevier Ltd. All rights reserved.

Controlling wildlife fungal disease spread: in vitro efficacy of disinfectants against Batrachochytrium dendrobatidis and Mucor amphibiorum.

PubMed

Webb, Rebecca; Philips, Annie; Speare, Rick; Connolly, Joanne; Berger, Lee

2012-06-13

Chytridiomycosis in amphibians, and mucormycosis in the platypus Ornithorhynchus anatinus and amphibians, are serious fungal diseases affecting these aquatic taxa. In Tasmania, Australia, the fungi that cause these diseases overlap in range along with Phytophthora cinnamomi (Pc), an invasive fungal plant pathogen. To identify disinfectants that may be useful to reduce anthropogenic spread of these fungi to uninfected wilderness areas, for example by bush walkers and forestry or fire-fighting operations, we tested 3 disinfectants and a fire-fighting foam against Mucor amphibiorum (Ma) and tested 1 disinfectant and the foam against Batrachochytrium dendrobatidis (Bd). Combining the present study with previous work we found Bd was more susceptible to all 4 chemicals than Ma. Phytoclean, a disinfectant used at 2 to 10% for 30 s to control Pc, killed cultures of Bd at 0.075% and Ma at 5%, when also applied for 30 s. The disinfectant F10sc was not effective against Ma at standard exposures, but previous work shows Bd is killed at 0.03% with a 1 min exposure. Path-X is effective against Bd at 0.001% with a 30 s exposure and killed Ma at 1% with a 5 min exposure. Forexpan S, a foam added to water at 0.1 to 1% to control forest fires, killed Bd but not Ma when used at 1% for 2 min. Therefore, Phytoclean and Path-X have broader efficacy, although Path-X has not been trialled against Pc. Interestingly a positive mating strain of Ma (from a platypus) was more resistant to disinfectants than a negative strain (from a frog). Current protocols against Pc that involve high concentrations (10%) of Phytoclean are likely to reduce spread of pathogenic wildlife fungi, which is important for protecting biodiversity.

Rapamycin Exerts Antifungal Activity In Vitro and In Vivo against Mucor circinelloides via FKBP12-Dependent Inhibition of Tor

PubMed Central

Bastidas, Robert J.; Shertz, Cecelia A.; Lee, Soo Chan; Heitman, Joseph

2012-01-01

The zygomycete Mucor circinelloides is an opportunistic fungal pathogen that commonly infects patients with malignancies, diabetes mellitus, and solid organ transplants. Despite the widespread use of antifungal therapy in the management of zygomycosis, the incidence of infections continues to rise among immunocompromised individuals. In this study, we established that the target and mechanism of antifungal action of the immunosuppressant rapamycin in M. circinelloides are mediated via conserved complexes with FKBP12 and a Tor homolog. We found that spontaneous mutations that disrupted conserved residues in FKBP12 conferred rapamycin and FK506 resistance. Disruption of the FKBP12-encoding gene, fkbA, also conferred rapamycin and FK506 resistance. Expression of M. circinelloides FKBP12 (McFKBP12) complemented a Saccharomyces cerevisiae mutant strain lacking FKBP12 to restore rapamycin sensitivity. Expression of the McTor FKBP12-rapamycin binding (FRB) domain conferred rapamycin resistance in S. cerevisiae, and McFKBP12 interacted in a rapamycin-dependent fashion with the McTor FRB domain in a yeast two-hybrid assay, validating McFKBP12 and McTor as conserved targets of rapamycin. We showed that in vitro, rapamycin exhibited potent growth inhibitory activity against M. circinelloides. In a Galleria mellonella model of systemic mucormycosis, rapamycin improved survival by 50%, suggesting that rapamycin and nonimmunosuppressive analogs have the potential to be developed as novel antifungal therapies for treatment of patients with mucormycosis. PMID:22210828

A non-canonical RNA degradation pathway suppresses RNAi-dependent epimutations in the human fungal pathogen Mucor circinelloides.

PubMed

Calo, Silvia; Nicolás, Francisco E; Lee, Soo Chan; Vila, Ana; Cervantes, Maria; Torres-Martinez, Santiago; Ruiz-Vazquez, Rosa M; Cardenas, Maria E; Heitman, Joseph

2017-03-01

Mucorales are a group of basal fungi that includes the casual agents of the human emerging disease mucormycosis. Recent studies revealed that these pathogens activate an RNAi-based pathway to rapidly generate drug-resistant epimutant strains when exposed to stressful compounds such as the antifungal drug FK506. To elucidate the molecular mechanism of this epimutation pathway, we performed a genetic analysis in Mucor circinelloides that revealed an inhibitory role for the non-canonical RdRP-dependent Dicer-independent silencing pathway, which is an RNAi-based mechanism involved in mRNA degradation that was recently identified. Thus, mutations that specifically block the mRNA degradation pathway, such as those in the genes r3b2 and rdrp3, enhance the production of drug resistant epimutants, similar to the phenotype previously described for mutation of the gene rdrp1. Our genetic analysis also revealed two new specific components of the epimutation pathway related to the quelling induced protein (qip) and a Sad-3-like helicase (rnhA), as mutations in these genes prevented formation of drug-resistant epimutants. Remarkably, drug-resistant epimutant production was notably increased in M. circinelloides f. circinelloides isolates from humans or other animal hosts. The host-pathogen interaction could be a stressful environment in which the phenotypic plasticity provided by the epimutant pathway might provide an advantage for these strains. These results evoke a model whereby balanced regulation of two different RNAi pathways is determined by the activation of the RNAi-dependent epimutant pathway under stress conditions, or its repression when the regular maintenance of the mRNA degradation pathway operates under non-stress conditions.

Protein Kinase A Regulatory Subunit Isoforms Regulate Growth and Differentiation in Mucor circinelloides: Essential Role of PKAR4

PubMed Central

Ocampo, J.; McCormack, B.; Navarro, E.; Moreno, S.; Garre, V.

2012-01-01

The protein kinase A (PKA) signaling pathway plays a role in regulating growth and differentiation in the dimorphic fungus Mucor circinelloides. PKA holoenzyme is comprised of two catalytic (C) and two regulatory (R) subunits. In M. circinelloides, four genes encode the PKAR1, PKAR2, PKAR3, and PKAR4 isoforms of R subunits. We have constructed null mutants and demonstrate that each isoform has a different role in growth and differentiation. The most striking finding is that pkaR4 is an essential gene, because only heterokaryons were obtained in knockout experiments. Heterokaryons with low levels of wild-type nuclei showed an impediment in the emission of the germ tube, suggesting a pivotal role of this gene in germ tube emergence. The remaining null strains showed different alterations in germ tube emergence, sporulation, and volume of the mother cell. The pkaR2 null mutant showed an accelerated germ tube emission and was the only mutant that germinated under anaerobic conditions when glycine was used as a nitrogen source, suggesting that pkaR2 participates in germ tube emergence by repressing it. From the measurement of the mRNA and protein levels of each isoform in the wild-type and knockout strains, it can be concluded that the expression of each subunit has its own mechanism of differential regulation. The PKAR1 and PKAR2 isoforms are posttranslationally modified by ubiquitylation, suggesting another regulation point in the specificity of the signal transduction. The results indicate that each R isoform has a different role in M. circinelloides physiology, controlling the dimorphism and contributing to the specificity of cyclic AMP (cAMP)-PKA pathway. PMID:22635921

Isolation and identification of fungi from a meju contaminated with aflatoxins.

PubMed

Jung, Yu Jung; Chung, Soo Hyun; Lee, Hyo Ku; Chun, Hyang Sook; Hong, Seung Beom

2012-12-01

A home-made meju sample contaminated naturally with aflatoxins was used for isolation of fungal strains. Overall, 230 fungal isolates were obtained on dichloran rosebengal chloramphenicol (DRBC) and dichloran 18% glycerol (DG18) agar plates. Morphological characteristics and molecular analysis of a partial beta-tubulin gene and the internal transcribed spacer (ITS) of rDNA were used for the identification of the isolates. The fungal isolates were divided into 7 genera: Aspergillus, Eurotium, Penicillium, Eupenicillium, Mucor, Lichtheimia, and Curvularia. Three strains from 56 isolates of the A. oryzae/flavus group were found to be aflatoxigenic A. flavus, by the presence of the aflatoxin biosynthesis genes and confirmatory aflatoxin production by high-performance liquid chromatography (HPLC). The predominant isolate from DRBC plates was A. oryzae (42 strains, 36.2%), whereas that from DG18 was A. candidus (61 strains, 53.5%). Out of the 230 isolates, the most common species was A. candidus (34.3%) followed by A. oryzae (22.2%), Mucor circinelloides (13.0%), P. polonicum (10.0%), A. tubingensis (4.8%), and L. ramosa (3.5%). A. flavus and E. chevalieri presented occurrence levels of 2.2%, respectively. The remaining isolates of A. unguis, P. oxalicum, Eupenicillium cinnamopurpureum, A. acidus, E. rubrum, P. chrysogenum, M. racemosus, and C. inaequalis had lower occurrence levels of < 2.0%.

The potential of soil fungi associated with potato rhizosphere to control root knot nematode (Meloidogyne spp.) on potato

NASA Astrophysics Data System (ADS)

Utari, E.; Lisnawita; Safni, I.; Lubis, K.; Tantawi, AR; Hasanuddin

2018-02-01

The root knot nematode (Meloidogyne spp.) is one of important pathogens on potato crops in North Sumatra, Indonesia. This nematode causes significant crop losses on potatoes directly and indirectly. The effect of fungal isolates (Trichoderma sp. 1, Mucor sp.1, Aspergillus sp. 2, Mucor sp. 2) that were isolated from rhizosphere of potato in North Sumatra were studied in green house experiments on the growth of potato and the reproduction of the nematode (Meloidogyne spp). The results showed that Trichoderma sp. 1 caused a significant gall reduction, while Mucor sp.1 and Mucor sp.2 could improve the growth of potato.

Early and Late Trisporoids Differentially Regulate β-Carotene Production and Gene Transcript Levels in the Mucoralean Fungi Blakeslea trispora and Mucor mucedo

PubMed Central

Sahadevan, Yamuna; Richter-Fecken, Mareike; Kaerger, Kerstin; Voigt, Kerstin

2013-01-01

The multistep cleavage of carotenoids in Mucorales during the sexual phase results in a cocktail of trisporic acid (C18) sex pheromones. We hypothesized that the C18 trisporoid intermediates have a specific regulatory function for sex pheromone production and carotenogenesis that varies with genus/species and vegetative and sexual phases of their life cycles. Real-time quantitative PCR kinetics determined for Blakeslea trispora displayed a very high transcript turnover in the gene for carotenoid cleavage dioxygenase, tsp3, during the sexual phase. An in vivo enzyme assay and chromatographic analysis led to the identification of β-apo-12′-carotenal as the first apocarotenoid involved in trisporic acid biosynthesis in B. trispora. Supplementation of C18 trisporoids, namely D'orenone, methyl trisporate C, and trisporin C, increased tsp3 transcripts in the plus compared to minus partners. Interestingly, the tsp1 gene, which is involved in trisporic acid biosynthesis, was downregulated compared to tsp3 irrespective of asexual or sexual phase. Only the minus partners of both B. trispora and Mucor mucedo had enhanced β-carotene production after treatment with C20 apocarotenoids, 15 different trisporoids, and their analogues. We conclude that the apocarotenoids and trisporoids influence gene transcription and metabolite production, depending upon the fungal strain, corresponding genus, and developmental phase, representing a “chemical dialect” during sexual communication. PMID:24056470

Dissecting metabolic behavior of lipid over-producing strain of Mucor circinelloides through genome-scale metabolic network and multi-level data integration.

PubMed

Vongsangnak, Wanwipa; Kingkaw, Amornthep; Yang, Junhuan; Song, Yuanda; Laoteng, Kobkul

2018-09-05

Lipid accumulation is an important cellular process of oleaginous microorganisms. To dissect metabolic behavior of oleaginous Zygomycetes, the lipid over-producing strain, Mucor circinelloides WJ11, was subjected for omics-scale analysis. The genome annotation was improved and used for construction of genome-scale metabolic network of WJ11 strain. Then, the quality of the metabolic network was enhanced by incorporating gene and protein expression data. In addition to the known oleaginous genes, our results showed a number of newly identified unique genes of WJ11 strain, which involved in central carbon metabolism, lipid, amino acid and nitrogen metabolisms. The systematic compilations indicated the additional metabolic routes with the involvement in supplying precursors (acetyl-CoA, NADPH and fatty acyl substrate) for fatty acid and lipid biosynthesis. Interestingly, amino acid metabolism played a substantial role in responsive mechanism of the fungal cells to nutrient imbalance circumstance through lipogenesis as the finding of reporter metabolites (l-methionine, l-glutamate, l-aspartate, l-asparagine and l-glutamine) at lipid-accumulating stage. The cooperative function of certain lipid-degrading enzymes at the particular growth stage was elucidated by integrating the metabolic networks with gene expression data. The unique feature of carotenoid biosynthetic route in WJ11 strain was also identified by protein domain analysis. Taken together, there were cross-functional metabolisms in regulating lipid biosynthesis and retaining high level of cellular lipids in the representative of lipid over-producing strains. Copyright © 2018 Elsevier B.V. All rights reserved.

Screening of Fungi for Biodegradation of Volatile Organic Compounds

DTIC Science & Technology

2004-04-20

more-robust alternative to bacteria in biofilter treatment. In the studies described herein, five fungal species, Exophiala lecanii-corni, Mucor ...degrade n-butyl acetate and methyl ethyl ketone but not benzene or p-xylene under the conditions tested. Mucor rouxii was able to use n-butyl...In the studies described herein, five fungal species, Exophiala lecanii-corni, Mucor rouxii (ATCC 44260), Phanerochaete chrysosporium (ATCC 24725

Fungal Burn Wound Infection

DTIC Science & Technology

1991-01-01

Aspergillus), Blasto- T he use of effective topical chemotherapeutic agents to myces (Candida), and Zygomycetes ( Mucor , Rhizopus).6 reduce...species, 18%; Mucor species and Rhizopus species, acetate in the morning and silver sulfadiazine in the evening. Prophy- 9.1%; and Microspora species and...sensitivity reports, and the patient’s sue, including one patient who required a hip disarticulation response. to control an invasive Mucor burn wound

Cause of Mortality in Thermally Injured Patients

DTIC Science & Technology

1993-01-01

11 patients, Cladosporium species from nine patients, Penicillium species from seven patients, and Phycomycetes ( mucor species and rhizopus species...infecting organisms were filamentous fungi, Aspergillus species in 16 and Mucor species in two, and in four patients C(nlida species were the causative...surface. Twelve, or 75 %, of the 16 patients with Aspergillus wound infections died but, somewhat surprisingly, both of the patients with mucor species

Lipases: Structure, Function and Applications in Biotransformations: A Descriptive Summary of an International Conference Held in Coventry (United Kingdom) on 16-18 July 1991

DTIC Science & Technology

1991-07-18

Basel) and Joseph D. Schrag (National Research Council of Canada, Montreal). These lecturers described the structures of the lipases from Mucor miehei...Geotrichum candidum, and human pancreas, respectively. Dr Huge- Jensen described the two forms of the Mucor enzyme that differed in their...hydrolysis). The crystal structure of the Mucor lipase showed that it consisted of an eight-stranded -sheet with an extended aX-helix at the N-terminus. The

[Intracardial fungal multiplication of order Mucor in an almost totally carbonised part of a male body found after ten days missing].

PubMed

Iannaccone, Silvia Farkašová; Klán, Jaroslav; Lamps, Laura W; Farkaš, Daniel; Švajdler Ml, Marián; Szabo, Miroslav

Determination of time of death belongs to the most difficult and also the most important issues for the medical examiners, especially those who deal with violent death. Besides the most frequently evaluated postmortal changes it is sometimes possible to perform the evaluation on the basis of less frequently observed findings. One of such findings is for example the fungal multiplication on the body or in the very close vicinity. Knowledge of moulds as well as information about their speed of growth should contribute to confirmation or negation of some information gained during police investigation. In this case report authors describe the macroscopically visible fungal intracardiac multiplication in heart chambers and aorta in an almost totally carbonised body which was missing for only ten days. Based on the molecular examination it was detected that the body belonged to the 64-year-old man who was repeatedly hospitalised in psychiatry for depression with suicidal tendencies. The last hospitalisation was six weeks before death and there was no organic disability. The cause of fire was a naked flame. The cause of death was burn injury or asphyxia. The almost total carbonisation did not allow to perform toxicological investigation. By histological investigation we found the presence of wide long non-septate moulds growing in the heart muscle, which belonged to the order Mucor. Since there was no obvious inflammatory response, we suppose their growth started on the congealed blood after death.

Characterization of an extracellular endopolygalacturonase from the saprobe Mucor ramosissimus Samutsevitsch and its action as trigger of defensive response in tropical plants.

PubMed

Marques, Maria Rita; Buckeridge, Marcos S; Braga, Marcia R; Dietrich, Sonia M C

2006-11-01

In recent years, interest in the ability of non-pathogenic microorganisms to induce resistance in plants has grown, particularly with respect to their use as environmentally safe controllers of plant disease. In this study, we investigated the capacity of Mucor ramosissimus Samutsevitsch to release pectinases able to degrade cell walls of Palicourea marcgravii St. Hil., a tropical forest native Rubiaceae on which the spores of this saprobic fungus have been found. The fungus was grown in liquid culture medium containing pectin as the sole carbon source and filtrates were analyzed for pectinase activity. An endopolygalacturonase was partially purified by ion exchange chromatography, gel filtration, and preparative isoelectrofocusing, and characterized. This enzyme was more active upon pectic substrates with a low degree of methyl esterification. The products of hydrolysis of different pectic substrates (including pectin from P. marcgravii) by the action of this endopolygalacturonase elicited to different extents the phytoalexin production in soybean cotyledons. Also, the enzyme itself and the products of its action on the pectic fraction of P. marcgravii elicited the production of defensive compounds in the leaves of the plant. These results suggest that, besides the role in recycling organic matter, saprobes may also play an important role in the induction of defensive mechanisms in wild plants by enhancing their non-specific resistance against pathogens. Furthermore, they set the stage for future studies on the role of saprobic fungi in inducing resistance of host plants to pathogens.

Mucormycosis Attributed Mortality: A Seven-year Review of Surgical and Medical Management

DTIC Science & Technology

2014-12-01

consideration for active wound surveillance. Specifically, fungal wound cultures diagnosed Mucor species in 7 out of 12 patients. Prior reports from...our institution have demonstrated mucor like morphol ogy was only able to be captured 40% of the time on fungal wound culture. Identification of these...theme of mucor infec tions is that of a large, open wound often with residual necrotic tissue regardless of the mechanism of injury that creates it, be

Construction of a Recyclable Genetic Marker and Serial Gene Deletions in the Human Pathogenic Mucorales Mucor circinelloides.

PubMed

Garcia, Alexis; Adedoyin, Gloria; Heitman, Joseph; Lee, Soo Chan

2017-07-05

Mucor circinelloides is a human pathogen, biofuel producer, and model system that belongs to a basal fungal lineage; however, the genetics of this fungus are limited. In contrast to ascomycetes and basidiomycetes, basal fungal lineages have been understudied. This may be caused by a lack of attention given to these fungi, as well as limited tools for genetic analysis. Nonetheless, the importance of these fungi as pathogens and model systems has increased. M. circinelloides is one of a few genetically tractable organisms in the basal fungi, but it is far from a robust genetic system when compared to model fungi in the subkingdom Dikarya. One problem is the organism is resistant to drugs utilized to select for dominant markers in other fungal transformation systems. Thus, we developed a blaster recyclable marker system by using the pyrG gene (encoding an orotidine-5'-phosphate decarboxylase, ortholog of URA3 in Saccharomyces cerevisiae ). A 237-bp fragment downstream of the pyrG gene was tandemly incorporated into the upstream region of the gene, resulting in construction of a pyrG-dpl237 marker. To test the functionality of the pyrG-dpl237 marker, we disrupted the carRP gene that is involved in carotenoid synthesis in pyrG - mutant background. The resulting carRP :: pyrG-dpl237 mutants exhibit a white colony phenotype due to lack of carotene, whereas wild type displays yellowish colonies. The pyrG marker was then successfully excised, generating carRP-dpl237 on 5-FOA medium. The mutants became auxotrophic and required uridine for growth. We then disrupted the calcineurin B regulatory subunit cnbR gene in the carRP :: dpl237 strain, generating mutants with the alleles carRP :: dpl237 and cnbR :: pyrG These results demonstrate that the recyclable marker system is fully functional, and therefore the pyrG-dpl237 marker can be used for sequential gene deletions in M. circinelloides . Copyright © 2017 Garcia et al.

Enzymatic bioremediation of polyaromatic hydrocarbons by fungal consortia enriched from petroleum contaminated soil and oil seeds.

PubMed

Balaji, V; Arulazhagan, P; Ebenezer, P

2014-05-01

The present study focuses on fungal strains capable of secreting extracellular enzymes by utilizing hydrocarbons present in the contaminated soil. Fungal strains were enriched from petroleum hydrocarbons contaminated soil samples collected from Chennai city, India. The potential fungi were isolated and screened for their enzyme secretion such as lipase, laccase, peroxidase and protease and also evaluated fungal enzyme mediated PAHs degradation. Total, 21 potential PAHs degrading fungi were isolated from PAHs contaminated soil, which belongs to 9 genera such as Aspergillus, Curvularia, Drechslera, Fusarium, Lasiodiplodia, Mucor Penicillium, Rhizopus, Trichoderma, and two oilseed-associated fungal genera such as Colletotrichum and Lasiodiplodia were used to test their efficacy in degradation of PAHs in polluted soil. Maximum lipase production was obtained with P. chrysogenum, M. racemosus and L. theobromae VBE1 under optimized cultural condition, which utilized PAHs in contaminated soil as sole carbon source. Fungal strains, P. chrysogenum, M. racemosus and L. theobromae VBE1, as consortia, used in the present study were capable of degrading branched alkane isoprenoids such as pristine (C17) and pyrene (C18) present in PAHs contaminated soil with high lipase production. The fungal consortia acts as potential candidate for bioremediation of PAHs contaminated environments.

Purification and characteristics of an inducible by polycyclic aromatic hydrocarbons NADP(+)-dependent naphthalenediol dehydrogenase (NDD) in Mucor circinelloides YR-1.

PubMed

Camacho-Morales, Reyna Lucero; Zazueta-Novoa, Vanesa; Casillas, Juana Lizbeth González; Ballesteros, Elizabeth Aranda; Bote, Juan Antonio Ocampo; Zazueta-Sandoval, Roberto

2014-05-01

We detected NADP(+)-dependent dihydrodiol dehydrogenase (DD) activity in a cell-free extract from Mucor circinelloides YR-1, after high-speed centrifugation. We analyzed the enzymatic activity in the cytosolic fraction by zymograms, as described previously, and eight different DD activity bands were revealed. Five constitutive DD activities (DD1-5) were present when glucose was used as carbon source and three inducible activities (NDD, PDD1 and PDD2) when aromatic hydrocarbon compounds were used. NDD activity was induced all of the aromatic hydrocarbon compounds. The highest DD activity inducer was naphthalene and the lowest was pyrene. One of the enzymes showed higher activity with cis-naphthalene-diol rather than with trans-nahthalenediol as a substrate. We purified this particular enzyme to homogeneity and found that it had an isoelectric point of 4.6. The molecular weight for the native protein was 197.4kDa and 49.03±0.5kDa for the monomer that conforms it, suggesting a homotetrameric structure for the complete enzyme. Polyclonal antibodies were raised against it and obtained. NDD activity was almost totally inhibited when antibodies were used at low concentrations, and in native immunoblots only one band, which corresponds to the activity band detected in the zymograms, could be detected. In denaturing PAGE immunoblots only one band was detected. This band corresponds to the purified protein band of 49kDa detected in SDS-PAGE gels. The other two inducible enzymes PDD1 and PDD2 were present only when phenanthrene was used as sole carbon source in the culture media. Copyright © 2014 Elsevier Inc. All rights reserved.

Lipase genes in Mucor circinelloides: identification, sub-cellular location, phylogenetic analysis and expression profiling during growth and lipid accumulation.

PubMed

Zan, Xinyi; Tang, Xin; Chu, Linfang; Zhao, Lina; Chen, Haiqin; Chen, Yong Q; Chen, Wei; Song, Yuanda

2016-10-01

Lipases or triacylglycerol hydrolases are widely spread in nature and are particularly common in the microbial world. The filamentous fungus Mucor circinelloides is a potential lipase producer, as it grows well in triacylglycerol-contained culture media. So far only one lipase from M. circinelloides has been characterized, while the majority of lipases remain unknown in this fungus. In the present study, 47 potential lipase genes in M. circinelloides WJ11 and 30 potential lipase genes in M. circinelloides CBS 277.49 were identified by extensive bioinformatics analysis. An overview of these lipases is presented, including several characteristics, sub-cellular location, phylogenetic analysis and expression profiling of the lipase genes during growth and lipid accumulation. All of these proteins contained the consensus sequence for a classical lipase (GXSXG motif) and were divided into four types including α/β-hydrolase_1, α/β-hydrolase_3, class_3 and GDSL lipase (GDSL) based on gene annotations. Phylogenetic analyses revealed that class_3 family and α/β-hydrolase_3 family were the conserved lipase family in M. circinelloides. Additionally, some lipases also contained a typical acyltransferase motif of H-(X) 4-D, and these lipases may play a dual role in lipid metabolism, catalyzing both lipid hydrolysis and transacylation reactions. The differential expression of all lipase genes were confirmed by quantitative real-time PCR, and the expression profiling were analyzed to predict the possible biological roles of these lipase genes in lipid metabolism in M. circinelloides. We preliminarily hypothesized that lipases may be involved in triacylglycerol degradation, phospholipid synthesis and beta-oxidation. Moreover, the results of sub-cellular localization, the presence of signal peptide and transcriptional analyses of lipase genes indicated that four lipase in WJ11 most likely belong to extracellular lipases with a signal peptide. These findings provide a platform

Genomic, proteomic, and biochemical analyses of oleaginous Mucor circinelloides: evaluating its capability in utilizing cellulolytic substrates for lipid production.

PubMed

Wei, Hui; Wang, Wei; Yarbrough, John M; Baker, John O; Laurens, Lieve; Van Wychen, Stefanie; Chen, Xiaowen; Taylor, Larry E; Xu, Qi; Himmel, Michael E; Zhang, Min

2013-01-01

Lipid production by oleaginous microorganisms is a promising route to produce raw material for the production of biodiesel. However, most of these organisms must be grown on sugars and agro-industrial wastes because they cannot directly utilize lignocellulosic substrates. We report the first comprehensive investigation of Mucor circinelloides, one of a few oleaginous fungi for which genome sequences are available, for its potential to assimilate cellulose and produce lipids. Our genomic analysis revealed the existence of genes encoding 13 endoglucanases (7 of them secretory), 3 β-D-glucosidases (2 of them secretory) and 243 other glycoside hydrolase (GH) proteins, but not genes for exoglucanases such as cellobiohydrolases (CBH) that are required for breakdown of cellulose to cellobiose. Analysis of the major PAGE gel bands of secretome proteins confirmed expression of two secretory endoglucanases and one β-D-glucosidase, along with a set of accessory cell wall-degrading enzymes and 11 proteins of unknown function. We found that M. circinelloides can grow on CMC (carboxymethyl cellulose) and cellobiose, confirming the enzymatic activities of endoglucanases and β-D-glucosidases, respectively. The data suggested that M. circinelloides could be made usable as a consolidated bioprocessing (CBP) strain by introducing a CBH (e.g. CBHI) into the microorganism. This proposal was validated by our demonstration that M. circinelloides growing on Avicel supplemented with CBHI produced about 33% of the lipid that was generated in glucose medium. Furthermore, fatty acid methyl ester (FAME) analysis showed that when growing on pre-saccharified Avicel substrates, it produced a higher proportion of C14 fatty acids, which has an interesting implication in that shorter fatty acid chains have characteristics that are ideal for use in jet fuel. This substrate-specific shift in FAME profile warrants further investigation.

Effect of the Filamentous Fungus Mucor circinelloides On The Development of Eggs of the Rumen Fluke Calicophoron daubneyi (Paramphistomidae).

PubMed

Arroyo, Fabián; Hernández, José A; Cazapal-Monteiro, Cristiana F; Pedreira, José; Sanchís, Jaime; Romasanta, Ángel; Sánchez-Andrade, Rita; Paz-Silva, Adolfo; Arias, María S

2017-06-01

Ruminants infected by Paramphistomidae flukes shed eggs in the feces, which pass through different stages in the environment until the infective stages (metacercariae) are reached. The activity of the soil fungus Mucor circinelloides on the development of eggs of the rumen fluke Calicophoron daubneyi was presently tested with 3 probes, i.e., in petri plates, feces, and an aqueous environment (tubes). The effect of the fungus was assessed by recording the numbers of undeveloped, nonviable, and embryonated eggs. Nonviable eggs were considered when vacuolization occurred, the inner structures were not clearly observed, the eggshell was broken, or the embryo inside was destroyed. By considering the ability of hyphae of M. circinelloides to develop in the presence of C. daubneyi eggs, attach to their surface, and penetrate and destroy the inner embryo, this ovicidal effect was classified as type 3. After a period of 50 days, the percentage of undeveloped eggs in the feces of infected cattle was 40%; furthermore, 27% of the eggs were nonviable, and 33% were embryonated (1 miracidium inside). The addition of 4 doses of M. circinelloides spores directly onto the feces resulted in 9-31% undeveloped eggs, 38-60% nonviable eggs, and 9-21% embryonated eggs, and no statistical significances were obtained among the different doses. Placing the eggs of C. daubneyi into an aqueous solution containing 10 7 spores of M. circinelloides/ml for 29 days resulted in 43% undeveloped eggs, 40% nonviable eggs, and 17% embryonated eggs, whereas in the controls, the percentages were 48%, 12%, and 40%, respectively. These data demonstrate the usefulness of the spores of the fungus M. circinelloides in limiting the development of the eggs of the trematode C. daubneyi.

Biosorption of Mercury (II) from Aqueous Solutions onto Fungal Biomass

PubMed Central

Martínez-Juárez, Víctor M.; Cárdenas-González, Juan F.; Torre-Bouscoulet, María Eugenia; Acosta-Rodríguez, Ismael

2012-01-01

The biosorption of mercury (II) on 14 fungal biomasses, Aspergillus flavus I–V, Aspergillus fumigatus I-II, Helminthosporium sp., Cladosporium sp., Mucor rouxii mutant, M. rouxii IM-80, Mucor sp 1 and 2, and Candida albicans, was studied in this work. It was found that the biomasses of the fungus M. rouxii IM-80, M. rouxii mutant, Mucor sp1, and Mucor sp 2 were very efficient removing the metal in solution, using dithizone, reaching the next percentage of removals: 95.3%, 88.7%, 80.4%, and 78.3%, respectively. The highest adsorption was obtained at pH 5.5, at 30°C after 24 hours of incubation, with 1 g/100 mL of fungal biomass. PMID:23028382

Oil and Hydrocarbon Spill Bioremediation Product and Application Technology

DTIC Science & Technology

1993-05-01

Kiebsiella Heiminthosporium Lactobacillus Mucor Leucothrix Oidiadendrumn Moraxella Paecylomyces Nocardia Phialophora Peptococcus Penicillium Pseudomonas...sludge’ Alkanes PsoudomonasPA rthroba eter, A cineeofbacter, yeasts, Penicillium sp., Cunninghamells blakesleearia, Absidiaglauca, Mucor spif n-Alkanes

Robust Phagocyte Recruitment Controls the Opportunistic Fungal Pathogen Mucor circinelloides in Innate Granulomas In Vivo.

PubMed

Inglesfield, Sarah; Jasiulewicz, Aleksandra; Hopwood, Matthew; Tyrrell, James; Youlden, George; Mazon-Moya, Maria; Millington, Owain R; Mostowy, Serge; Jabbari, Sara; Voelz, Kerstin

2018-03-27

Mucormycosis is an emerging fungal infection with extremely high mortality rates in patients with defects in their innate immune response, specifically in functions mediated through phagocytes. However, we currently have a limited understanding of the molecular and cellular interactions between these innate immune effectors and mucormycete spores during the early immune response. Here, the early events of innate immune recruitment in response to infection by Mucor circinelloides spores are modeled by a combined in silico modeling approach and real-time in vivo microscopy. Phagocytes are rapidly recruited to the site of infection in a zebrafish larval model of mucormycosis. This robust early recruitment protects from disease onset in vivo In silico analysis identified that protection is dependent on the number of phagocytes at the infection site, but not the speed of recruitment. The mathematical model highlights the role of proinflammatory signals for phagocyte recruitment and the importance of inhibition of spore germination for protection from active fungal disease. These in silico data are supported by an in vivo lack of fungal spore killing and lack of reactive oxygen burst, which together result in latent fungal infection. During this latent stage of infection, spores are controlled in innate granulomas in vivo Disease can be reactivated by immunosuppression. Together, these data represent the first in vivo real-time analysis of innate granuloma formation during the early stages of a fungal infection. The results highlight a potential latent stage during mucormycosis that should urgently be considered for clinical management of patients. IMPORTANCE Mucormycosis is a dramatic fungal infection frequently leading to the death of patients. We know little about the immune response to the fungus causing this infection, although evidence points toward defects in early immune events after infection. Here, we dissect this early immune response to infectious fungal

Comparison of Biochemical Activities between High and Low Lipid-Producing Strains of Mucor circinelloides: An Explanation for the High Oleaginicity of Strain WJ11.

PubMed

Tang, Xin; Chen, Haiqin; Chen, Yong Q; Chen, Wei; Garre, Victoriano; Song, Yuanda; Ratledge, Colin

2015-01-01

The oleaginous fungus, Mucor circinelloides, is one of few fungi that produce high amounts of γ-linolenic acid (GLA); however, it usually only produces <25% lipid. Nevertheless, a new strain (WJ11) isolated in this laboratory can produce lipid up to 36% (w/w) cell dry weight (CDW). We have investigated the potential mechanism of high lipid accumulation in M. circinelloides WJ11 by comparative biochemical analysis with a low lipid-producing strain, M. circinelloides CBS 277.49, which accumulates less than 15% (w/w) lipid. M. circinelloides WJ11 produced more cell mass than that of strain CBS 277.49, although with slower glucose consumption. In the lipid accumulation phase, activities of glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in strain WJ11 were greater than in CBS 277.49 by 46% and 17%, respectively, and therefore may provide more NADPH for fatty acid biosynthesis. The activities of NAD+:isocitrate dehydrogenase and NADP+:isocitrate dehydrogenase, however, were 43% and 54%, respectively, lower in WJ11 than in CBS 277.49 and may retard the tricarboxylic acid cycle and thereby provide more substrate for ATP:citrate lyase (ACL) to produce acetyl-CoA. Also, the activities of ACL and fatty acid synthase in the high lipid-producing strain, WJ11, were 25% and 56%, respectively, greater than in strain CBS 277.49. These enzymes may therefore cooperatively regulate the fatty acid biosynthesis in these two strains.

Isolation of Mucorales from processed maize (Zea mays L.) and screening for protease activity

PubMed Central

de Azevedo Santiago, André Luiz Cabral Monteiro; de Souza Motta, Cristina Maria

2008-01-01

Mucorales were isolated from maize flour, corn meal and cooked cornflakes using surface and depth plate methods. Rhizopus oryzae, Circinella muscae, Mucor subtilissimus, Mucor hiemalis f. hiemalis, Syncephalastrum racemosum, Rhizopus microsporus var. chinensis and Absidia cylindrospora showed protease activity. PMID:24031292

Volatile organic compounds emitted by filamentous fungi isolated from flooded homes after Hurricane Sandy show toxicity in a Drosophila bioassay.

PubMed

Zhao, G; Yin, G; Inamdar, A A; Luo, J; Zhang, N; Yang, I; Buckley, B; Bennett, J W

2017-05-01

Superstorm Sandy provided an opportunity to study filamentous fungi (molds) associated with winter storm damage. We collected 36 morphologically distinct fungal isolates from flooded buildings. By combining traditional morphological and cultural characters with an analysis of ITS sequences (the fungal DNA barcode), we identified 24 fungal species that belong to eight genera: Penicillium (11 species), Fusarium (four species), Aspergillus (three species), Trichoderma (two species), and one species each of Metarhizium, Mucor, Pestalotiopsis, and Umbelopsis. Then, we used a Drosophila larval assay to assess possible toxicity of volatile organic compounds (VOCs) emitted by these molds. When cultured in a shared atmosphere with growing cultures of molds isolated after Hurricane Sandy, larval toxicity ranged from 15 to 80%. VOCs from Aspergillus niger 129B were the most toxic yielding 80% mortality to Drosophila after 12 days. The VOCs from Trichoderma longibrachiatum 117, Mucor racemosus 138a, and Metarhizium anisopliae 124 were relatively non-toxigenic. A preliminary analysis of VOCs was conducted using solid-phase microextraction-gas chromatography-mass spectrometry from two of the most toxic, two of the least toxic, and two species of intermediate toxicity. The more toxic molds produced higher concentrations of 1-octen-3-ol, 3-octanone, 3-octanol, 2-octen-1-ol, and 2-nonanone; while the less toxic molds produced more 3-methyl-1-butanol and 2-methyl-1-propanol, or an overall lower amount of volatiles. Our data support the hypothesis that at certain concentrations, some VOCs emitted by indoor molds are toxigenic. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Using aquatic fungi for pharmaceutical bioremediation: Uptake of acetaminophen by Mucor hiemalis does not result in an enzymatic oxidative stress response.

PubMed

Esterhuizen-Londt, Maranda; Schwartz, Katrin; Pflugmacher, Stephan

2016-10-01

The increasing anthropogenic pollution of aquatic environments and fresh water scarcity worldwide have prompted the development of low-cost and effective water treatment alternatives. One example of a highly released anthropogenic xenobiotics is acetaminophen (APAP), which has been detected in surface waters at concentrations as high as 5 μg L(-1). To date, traditional water treatment plants were unable to remove all pharmaceutical xenobiotics and as in the case with APAP, the breakdown products are toxic. Phytoremediation has proved to remove xenobiotics efficiently producing no toxic breakdown products, however, they are often restrained in their application range. Therefore, it was necessary to find alternate remediation tools to extend and complement the application ranges of existing bioremediation techniques. With the success of mycoremediation as well as the adaptability of fungi, Mucor hiemalis was investigated in terms of its APAP uptake capabilities. The investigation included the examination of concentration- and time-dependent uptake studies to examine the effects of each of these parameters independently. Additionally, the extracellular peroxidase activity of M. hiemalis was measured with exposure to APAP to evaluate possible breakdown and the antioxidative stress enzymes, catalase, glutathione peroxidase, and glutathione reductase, were assayed to investigate whether APAP caused oxidative stress. The results showed that M. hiemalis was able to internalize between 1 and 2 μg APAP per g dried fungal biomass when exposed to 5, 10, 50 and 100 ng mL(-1) APAP for 24-48 h, but not beyond this time frame. Further, exposure to APAP did not result in elevated extracellular peroxidase activity or oxidative stress. The findings led to the conclusion that M. hiemalis could be integrated in bioremediation systems, for short-term degradation at low concentrations of APAP with effective management. Copyright © 2016 British Mycological Society. Published by

The Effect of Hyperbaric Oxygen on the Growth of Rhizopus nigricans.

DTIC Science & Technology

1980-12-01

identify by block number) Impression Chamber Medical Mycology Oxygen Therapy (ompression Therapy Mucor Phycomycosis hvp)erbaric Medicine Mucormycosis...rado. 3. Cai rnev, W. J. 1980. The effect of hyperbaric oxygen on the growth of Mucor sp. and Aspergillus fumigLatus. Technical Report FJSRL-TR- 80

Incidence of Systemic Fungal Infection and Related Mortality Following Severe Burns

DTIC Science & Technology

2008-01-01

had Mucor , 7 patients had Fusarium, 3 patients had Alternaria, 4 patients had Penicillium, 2 patients had Cladosporium, 2 patients had Trichsoporon, 1...Aspergillus in these 13 cases included Candida in 3 patients, Mucor in 2 patients, and Fusarium in 2 patients. The presence of fungal elements in autopsied

Detection of Fungi from an Indoor Environment using Loop-mediated Isothermal Amplification (LAMP) Method.

PubMed

Nakayama, Takako; Yamazaki, Takashi; Yo, Ayaka; Tone, Kazuya; Mahdi Alshahni, Mohamed; Fujisaki, Ryuichi; Makimura, Koichi

2017-01-01

　Loop-mediated isothermal amplification (LAMP) is a useful DNA detection method with high specificity and sensitivity. The LAMP reaction is carried out within a short time at a constant temperature without the need for thermal cycling. We developed a LAMP primer set for detecting a wide range of fungi by aligning the sequences of the large subunit ribosomal RNA gene of Candida albicans (Ascomycota), Cryptococcus neoformans (Basidiomycota), and Mucor racemosus (Mucorales). The threshold of C. albicans rDNA as template with our LAMP primer set was in the range of 10-100 copies per a reaction. In this study, we evaluated the correlation between colony forming units (CFU) and LAMP detection rate using the LAMP method for environmental fungi. The LAMP method should be a useful means of detecting fungi in indoor environments, disaster areas, or even in confined manned spacecraft to prevent allergies or infections caused by fungi.

Chemical composition and antimicrobial activities of essential oil from Wedelia urticifolia growing wild in Hunan Province, China.

PubMed

Hu, Junpeng; Jia, Mengmeng; Zhu, Liang

2018-04-18

The essential oil obtained from Wedelia urticifolia growing in Hunan Province, China, was analyzed for the first time by capillary GC and GC-MS. A total of 67 constituents, representing 98.68% in essential oil were identified. The major constituents of the oil were: α-pinene (8.85%), limonene (6.38%), carvacrol (6.15%), caryophyllene (6.08%), spathulenol (5.49%), sabinene (5.36%), camphor (4.34%). Antimicrobial potential of oil against bacterial strains (Pseudomonas aeruginosa, Escherichia coli and Bacillus subtilis, and Staphylococcus aureus), yeast strains (Hansenula anomala and Saccharomy cescerevisiae) and molds (Aspergillus niger, Chaetomium globosum, Mucor racemosus, and Monascus anka) was determined by disc diffusion method and broth micro dilution method, respectively. The oil exhibited promising antimicrobial effect as a diameter of zones of inhibition (16.8-24.9 mm). Minimum inhibitory concentration values of oil were ranged 62.5-1000 μg/mL.

[Clinical analyses of the diagnosis and treatment of invasive fungal rhinosinusitis: report of 14 cases].

PubMed

Shi, G G; Shi, L; Zhang, Z Y; Wan, Y Z; Li, B; Yu, L; Zhang, E P; Ju, H S; He, M Q; Ji, H Z

2016-08-07

Through the retrospective analysis of the clinical data in 14 cases of invasive fungal rhinosinusitis (IFRS), the clinical characteristics, diagnosis and treatment of this disease were evaluated. Fourteen clinically confirmed cases of IFRS since January 2008 to October 2015 were evaluated.collected, the clinical features, diagnosis, treatment and prognosis were analyzed to obtain a more comprehensive understanding for clinical reference. Fourteen patients were confirmed by pathological examination as IFRS, including 9 cases of aspergillus, 4 cases of mucor, and 1 case of rhinocerebral zygomycosis; including 5 cases of acute IFRS, 9 cases of chronic IFRS. All patients were treated with endoscopic surgery and intravenous antifungal therapy. Nine cases of chronic IFRS (including 1 case of mucor, 7 cases of aspergillus and 1 case of rhinocerebral zygomycosis) were cured, but the vision loss, diplopia or blindness, hard palate perforation remained. Five cases of acute IFRS included 3 cases of mucor and 2 cases of aspergillus. Among the 3 cases of mucor, 2 cases were died and 1 case was cured. Among the 2 cases of aspergillus, 1 patient was cured and the other patient died of electrolyte disorder after discharge from hospital. Patients with IFRS usually have diabetes. After the active surgical cleaning of lesion tissue and the systematic antifungal treatment with adequate dosage, these patients would have a better result. IFRS caused by mucor is ofen dangerous.

Associations between fungal species and water-damaged building materials.

PubMed

Andersen, Birgitte; Frisvad, Jens C; Søndergaard, Ib; Rasmussen, Ib S; Larsen, Lisbeth S

2011-06-01

Fungal growth in damp or water-damaged buildings worldwide is an increasing problem, which has adverse effects on both the occupants and the buildings. Air sampling alone in moldy buildings does not reveal the full diversity of fungal species growing on building materials. One aim of this study was to estimate the qualitative and quantitative diversity of fungi growing on damp or water-damaged building materials. Another was to determine if associations exist between the most commonly found fungal species and different types of materials. More than 5,300 surface samples were taken by means of V8 contact plates from materials with visible fungal growth. Fungal identifications and information on building material components were analyzed using multivariate statistic methods to determine associations between fungi and material components. The results confirmed that Penicillium chrysogenum and Aspergillus versicolor are the most common fungal species in water-damaged buildings. The results also showed Chaetomium spp., Acremonium spp., and Ulocladium spp. to be very common on damp building materials. Analyses show that associated mycobiotas exist on different building materials. Associations were found between (i) Acremonium spp., Penicillium chrysogenum, Stachybotrys spp., Ulocladium spp., and gypsum and wallpaper, (ii) Arthrinium phaeospermum, Aureobasidium pullulans, Cladosporium herbarum, Trichoderma spp., yeasts, and different types of wood and plywood, and (iii) Aspergillus fumigatus, Aspergillus melleus, Aspergillus niger, Aspergillus ochraceus, Chaetomium spp., Mucor racemosus, Mucor spinosus, and concrete and other floor-related materials. These results can be used to develop new and resistant building materials and relevant allergen extracts and to help focus research on relevant mycotoxins, microbial volatile organic compounds (MVOCs), and microparticles released into the indoor environment.

Associations between Fungal Species and Water-Damaged Building Materials ▿

PubMed Central

Andersen, Birgitte; Frisvad, Jens C.; Søndergaard, Ib; Rasmussen, Ib S.; Larsen, Lisbeth S.

2011-01-01

Fungal growth in damp or water-damaged buildings worldwide is an increasing problem, which has adverse effects on both the occupants and the buildings. Air sampling alone in moldy buildings does not reveal the full diversity of fungal species growing on building materials. One aim of this study was to estimate the qualitative and quantitative diversity of fungi growing on damp or water-damaged building materials. Another was to determine if associations exist between the most commonly found fungal species and different types of materials. More than 5,300 surface samples were taken by means of V8 contact plates from materials with visible fungal growth. Fungal identifications and information on building material components were analyzed using multivariate statistic methods to determine associations between fungi and material components. The results confirmed that Penicillium chrysogenum and Aspergillus versicolor are the most common fungal species in water-damaged buildings. The results also showed Chaetomium spp., Acremonium spp., and Ulocladium spp. to be very common on damp building materials. Analyses show that associated mycobiotas exist on different building materials. Associations were found between (i) Acremonium spp., Penicillium chrysogenum, Stachybotrys spp., Ulocladium spp., and gypsum and wallpaper, (ii) Arthrinium phaeospermum, Aureobasidium pullulans, Cladosporium herbarum, Trichoderma spp., yeasts, and different types of wood and plywood, and (iii) Aspergillus fumigatus, Aspergillus melleus, Aspergillus niger, Aspergillus ochraceus, Chaetomium spp., Mucor racemosus, Mucor spinosus, and concrete and other floor-related materials. These results can be used to develop new and resistant building materials and relevant allergen extracts and to help focus research on relevant mycotoxins, microbial volatile organic compounds (MVOCs), and microparticles released into the indoor environment. PMID:21531835

A mycological investigation of phane, an edible caterpillar of an emperor moth, Imbrasia belina.

PubMed

Simpanya, M F; Allotey, J; Mpuchane, S F

2000-01-01

Phane worm (an edible larval stage of the emperor moth Imbrasia belina Westwood) is an important food source, and its harvesting is an economic activity in rural Botswana. When the larva is feeding on leaves and later during processing, phane gets contaminated with fungi from the leaves and soil. We examined 73 jars, each containing approximately 608 g (+/-0.25 g) of processed phane stored under laboratory conditions (temperature range 20 to 24 degrees C and 50 to 80% relative humidity) and combined intestinal contents of five phane squeezed into each of 74 Duran bottles for fungi. Ninety seven percent of 74 samples of intestinal contents and 57.5% of 73 laboratory-stored phane were positive for either molds and/or yeasts. Yeast population in intestinal contents ranged from 2 x 10(1) CFU/g to 5 x 10(3) CFU/g, whereas molds ranged from 1 x 10(1) CFU/g to 2 x 10(2) CFU/g. Laboratory-stored phane had a mold population of 1 x 10(2) CFU/g to 6 x 10(5) CFU/g. Species of Chaetomium 13.8%, Aspergillus 12.4%, Fusarium 5.5%, and Mucor racemosus 4.1% were the most prevalent in intestinal contents of phane, whereas Aspergillus 42.1%, Penicillium 33.9%, and Mucorales 5.7% were predominant in laboratory-stored phane. The important mycotoxigenic fungi A. flavus, A. parasiticus, A. ochraceus, P. aurantiogriseum, P. citrinum, and P. verrucosum were isolated mainly from the laboratory-stored phane. The genera isolated from both intestinal phane contents and laboratory-stored phane were Alternaria, Aspergillus, Chaetomium, Drechslera, Fusarium, Mucor, Phoma, and Penicillium, suggesting recontamination of phane during drying and storage.

Efficacy of a coating composed of chitosan from Mucor circinelloides and carvacrol to control Aspergillus flavus and the quality of cherry tomato fruits

PubMed Central

de Souza, Evandro L.; Sales, Camila V.; de Oliveira, Carlos E. V.; Lopes, Laênia A. A.; da Conceição, Maria L.; Berger, Lúcia R. R.; Stamford, Thayza C. M.

2015-01-01

Cherry tomato (Lycopersicon esculentum Mill) fruits are susceptible to contamination by Aspergillus flavus, which may cause the development of fruit rot and significant postharvest losses. Currently there are significant drawbacks for the use of synthetic fungicides to control pathogenic fungi in tomato fruits, and it has increased the interest in exploring new alternatives to control the occurrence of fungal infections in these fruits. This study evaluated the efficacy of chitosan (CHI) from Mucor circinelloides in combination with carvacrol (CAR) in inhibiting A. flavus in laboratory media and as a coating on cherry tomato fruits (25°C, 12 days and 12°C, 24 days). During a period of storage, the effect of coatings composed of CHI and CAR on autochthonous microflora, as well as on some quality characteristics of the fruits such as weight loss, color, firmness, soluble solids, and titratable acidity was evaluated. CHI and CAR displayed MIC valuesof 7.5 mg/mL and 10 μL/mL, respectively, against A. flavus. The combined application of CHI (7.5 or 3.75 mg/mL) and CAR (5 or 2.5 μL/mL) strongly inhibited the mycelial growth and spore germination of A. flavus. The coating composed of CHI (3.75 mg/mL) and CAR (2.5 or 1.25 μL/mL) inhibited the growth of A. flavus in artificially contaminated fruits, as well as the native fungal microflora of the fruits stored at room or low temperature. The application of the tested coatings preserved the quality of cherry tomato fruits as measured by some physicochemical attributes. From this, composite coatings containing CHI and CAR offer a promising alternative to control postharvest infection caused by A. flavus or native fungal microflora in fresh cherry tomato fruits without negatively affecting their quality over storage. PMID:26257717

Activity of selected essential oils on spoiling fungi cultured from Marzolino cheese.

PubMed

Nardoni, Simona; D'Ascenzi, Carlo; Caracciolo, Irene; Mannaioni, Gaia; Papini, Roberto Amerigo; Pistelli, Luisa; Najar, Basma; Mancianti, Francesca

2018-06-20

Microscopic fungi can be present on a variety of foodstuff, including cheese. They can be responsible for fungal spoilage, causing sensory changes making food unacceptable for human consumption, and posing severe health concerns. Furthermore, some of these organisms are able to resist antimicrobial preservatives provided for by law. Antifungal activity of 15 chemically defined EOs, alone and in mixture, were checked by a microdilution test against isolates of Penicillium funiculosum and Mucor racemosus cultured from rinds of Marzolino, a typical Italian fresh pecorino cheese. Origanum vulgare yielded the lowest MIC values, followed by Salvia sclarea, Ocimum basilicum and Cymbopogon citratus, while Citrus paradisi and Citrus limon were not active. All mixtures showed antifungal activity at lower concentration with respect to MIC values of each EO component, when not in combination. This study is the first to describe the setting up of EOs mixtures to limit spoiling moulds.

Stress response in medically important Mucorales.

PubMed

Singh, Pankaj; Paul, Saikat; Shivaprakash, M Rudramurthy; Chakrabarti, Arunaloke; Ghosh, Anup K

2016-10-01

Mucorales are saprobes, ubiquitously distributed and able to infect a heterogeneous population of human hosts. The fungi require robust stress responses to survive in human host. We tested the growth of Mucorales in the presence of different abiotic stress. Eight pathogenic species of Mucorales, including Rhizopus arrhizus, Rhizopus microsporus, Rhizomucor pusillus, Apophysomyces elegans, Licthemia corymbifera, Cunninghamella bertholletiae, Syncephalastrum racemosum and Mucor racemosus, were exposed to different stress inducers: osmotic (sodium chloride and d-sorbitol), oxidative (hydrogen peroxide and menadione), pH, cell wall and metal ions (Cu, Zn, Fe and Mg). Wide variation in stress responses was noted: R. arrhizus showed maximum resistance to both osmotic and oxidative stresses, whereas R. pusillus and M. indicus were relatively sensitive. Rhizopus arrhizus and R. microsporus showed maximum resistance to alkaline pH, whereas C. bertholletiae, L. corymbifera, M. racemosus and A. elegans were resistant to acidic pH. Maximum tolerance was noted in R. microsporus to Cu, R. microsporus and R. arrhizus to Fe and C. bertholletiae to Zn. In contrast, L. corymbifera, A. elegans and M. indicus were sensitive to Cu, Zn and Fe respectively. In conclusion, R. arrhizus showed high stress tolerance in comparison to other species of Mucorales, and this could be the possible reason for high pathogenic potential of this fungi. © 2016 Blackwell Verlag GmbH.

Morphological evolution of various fungal species in the presence and absence of aluminum oxide microparticles: Comparative and quantitative insights into microparticle-enhanced cultivation (MPEC).

PubMed

Kowalska, Anna; Boruta, Tomasz; Bizukojć, Marcin

2018-03-05

The application of microparticle-enhanced cultivation (MPEC) is an attractive method to control mycelial morphology, and thus enhance the production of metabolites and enzymes in the submerged cultivations of filamentous fungi. Unfortunately, most literature data deals with the spore-agglomerating species like aspergilli. Therefore, the detailed quantitative study of the morphological evolution of four different fungal species (Aspergillus terreus, Penicillium rubens, Chaetomium globosum, and Mucor racemosus) based on the digital analysis of microscopic images was presented in this paper. In accordance with the current knowledge, these species exhibit different mechanisms of agglomerates formation. The standard submerged shake flask cultivations (as a reference) and MPEC involving 10 μm aluminum oxide microparticles (6 g·L -1 ) were performed. The morphological parameters, including mean projected area, elongation, roughness, and morphology number were determined for the mycelial objects within the first 24 hr of growth. It occurred that heretofore observed and widely discussed effect of microparticles on fungi, namely the decrease in pellet size, was not observed for the species whose pellet formation mechanism is different from spore agglomeration. In the MPEC, C. globosum developed core-shell pellets, and M. racemosus, a nonagglomerative species, formed the relatively larger, compared to standard cultures, pellets with distinct cores. © 2018 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

Antifungal Susceptibility and Phylogeny of Opportunistic Members of the Order Mucorales

PubMed Central

Vitale, Roxana G.; de Hoog, G. Sybren; Schwarz, Patrick; Dannaoui, Eric; Deng, Shuwen; Machouart, Marie; Voigt, Kerstin; van de Sande, Wendy W. J.; Dolatabadi, Somayeh; Meis, Jacques F.

2012-01-01

The in vitro susceptibilities of 66 molecularly identified strains of the Mucorales to eight antifungals (amphotericin B, terbinafine, itraconazole, posaconazole, voriconazole, caspofungin, micafungin, and 5-fluorocytosine) were tested. Molecular phylogeny was reconstructed based on the nuclear ribosomal large subunit to reveal taxon-specific susceptibility profiles. The impressive phylogenetic diversity of the Mucorales was reflected in susceptibilities differing at family, genus, and species levels. Amphotericin B was the most active drug, though somewhat less against Rhizopus and Cunninghamella species. Posaconazole was the second most effective antifungal agent but showed reduced activity in Mucor and Cunninghamella strains, while voriconazole lacked in vitro activity for most strains. Genera attributed to the Mucoraceae exhibited a wide range of MICs for posaconazole, itraconazole, and terbinafine and included resistant strains. Cunninghamella also comprised strains resistant to all azoles tested but was fully susceptible to terbinafine. In contrast, the Lichtheimiaceae completely lacked strains with reduced susceptibility for these antifungals. Syncephalastrum species exhibited susceptibility profiles similar to those of the Lichtheimiaceae. Mucor species were more resistant to azoles than Rhizopus species. Species-specific responses were obtained for terbinafine where only Rhizopus arrhizus and Mucor circinelloides were resistant. Complete or vast resistance was observed for 5-fluorocytosine, caspofungin, and micafungin. Intraspecific variability of in vitro susceptibility was found in all genera tested but was especially high in Mucor and Rhizopus for azoles and terbinafine. Accurate molecular identification of etiologic agents is compulsory to predict therapy outcome. For species of critical genera such as Mucor and Rhizopus, exhibiting high intraspecific variation, susceptibility testing before the onset of therapy is recommended. PMID:22075600

Antifungal susceptibility and phylogeny of opportunistic members of the order mucorales.

PubMed

Vitale, Roxana G; de Hoog, G Sybren; Schwarz, Patrick; Dannaoui, Eric; Deng, Shuwen; Machouart, Marie; Voigt, Kerstin; van de Sande, Wendy W J; Dolatabadi, Somayeh; Meis, Jacques F; Walther, Grit

2012-01-01

The in vitro susceptibilities of 66 molecularly identified strains of the Mucorales to eight antifungals (amphotericin B, terbinafine, itraconazole, posaconazole, voriconazole, caspofungin, micafungin, and 5-fluorocytosine) were tested. Molecular phylogeny was reconstructed based on the nuclear ribosomal large subunit to reveal taxon-specific susceptibility profiles. The impressive phylogenetic diversity of the Mucorales was reflected in susceptibilities differing at family, genus, and species levels. Amphotericin B was the most active drug, though somewhat less against Rhizopus and Cunninghamella species. Posaconazole was the second most effective antifungal agent but showed reduced activity in Mucor and Cunninghamella strains, while voriconazole lacked in vitro activity for most strains. Genera attributed to the Mucoraceae exhibited a wide range of MICs for posaconazole, itraconazole, and terbinafine and included resistant strains. Cunninghamella also comprised strains resistant to all azoles tested but was fully susceptible to terbinafine. In contrast, the Lichtheimiaceae completely lacked strains with reduced susceptibility for these antifungals. Syncephalastrum species exhibited susceptibility profiles similar to those of the Lichtheimiaceae. Mucor species were more resistant to azoles than Rhizopus species. Species-specific responses were obtained for terbinafine where only Rhizopus arrhizus and Mucor circinelloides were resistant. Complete or vast resistance was observed for 5-fluorocytosine, caspofungin, and micafungin. Intraspecific variability of in vitro susceptibility was found in all genera tested but was especially high in Mucor and Rhizopus for azoles and terbinafine. Accurate molecular identification of etiologic agents is compulsory to predict therapy outcome. For species of critical genera such as Mucor and Rhizopus, exhibiting high intraspecific variation, susceptibility testing before the onset of therapy is recommended.

DNA barcoding in Mucorales: an inventory of biodiversity.

PubMed

Walther, G; Pawłowska, J; Alastruey-Izquierdo, A; Wrzosek, M; Rodriguez-Tudela, J L; Dolatabadi, S; Chakrabarti, A; de Hoog, G S

2013-06-01

The order Mucorales comprises predominantly fast-growing saprotrophic fungi, some of which are used for the fermentation of foodstuffs but it also includes species known to cause infections in patients with severe immune or metabolic impairments. To inventory biodiversity in Mucorales ITS barcodes of 668 strains in 203 taxa were generated covering more than two thirds of the recognised species. Using the ITS sequences, Molecular Operational Taxonomic Units were defined by a similarity threshold of 99 %. An LSU sequence was generated for each unit as well. Analysis of the LSU sequences revealed that conventional phenotypic classifications of the Mucoraceae are highly artificial. The LSU- and ITS-based trees suggest that characters, such as rhizoids and sporangiola, traditionally used in mucoralean taxonomy are plesiomorphic traits. The ITS region turned out to be an appropriate barcoding marker in Mucorales. It could be sequenced directly in 82 % of the strains and its variability was sufficient to resolve most of the morphospecies. Molecular identification turned out to be problematic only for the species complexes of Mucor circinelloides, M. flavus, M. piriformis and Zygorhynchus moelleri. As many as 12 possibly undescribed species were detected. Intraspecific variability differed widely among mucorealean species ranging from 0 % in Backusella circina to 13.3 % in Cunninghamella echinulata. A high proportion of clinical strains was included for molecular identification. Clinical isolates of Cunninghamella elegans were identified molecularly for the first time. As a result of the phylogenetic analyses several taxonomic and nomenclatural changes became necessary. The genus Backusella was emended to include all species with transitorily recurved sporangiophores. Since this matched molecular data all Mucor species possessing this character were transferred to Backusella. The genus Zygorhynchus was shown to be polyphyletic based on ITS and LSU data. Consequently

Efficient anchoring of alien chromosome segments introgressed into bread wheat by new Leymus racemosus genome-based markers.

PubMed

Edet, Offiong Ukpong; Kim, June-Sik; Okamoto, Masanori; Hanada, Kousuke; Takeda, Tomoyuki; Kishii, Masahiro; Gorafi, Yasir Serag Alnor; Tsujimoto, Hisashi

2018-03-27

The tertiary gene pool of bread wheat, to which Leymus racemosus belongs, has remained underutilized due to the current limited genomic resources of the species that constitute it. Continuous enrichment of public databases with useful information regarding these species is, therefore, needed to provide insights on their genome structures and aid successful utilization of their genes to develop improved wheat cultivars for effective management of environmental stresses. We generated de novo DNA and mRNA sequence information of L. racemosus and developed 110 polymorphic PCR-based markers from the data, and to complement the PCR markers, DArT-seq genotyping was applied to develop additional 9990 SNP markers. Approximately 52% of all the markers enabled us to clearly genotype 22 wheat-L. racemosus chromosome introgression lines, and L. racemosus chromosome-specific markers were highly efficient in detailed characterization of the translocation and recombination lines analyzed. A further analysis revealed remarkable transferability of the PCR markers to three other important Triticeae perennial species: L. mollis, Psathyrostachys huashanica and Elymus ciliaris, indicating their suitability for characterizing wheat-alien chromosome introgressions carrying chromosomes of these genomes. The efficiency of the markers in characterizing wheat-L. racemosus chromosome introgression lines proves their reliability, and their high transferability further broadens their scope of application. This is the first report on sequencing and development of markers from L. racemosus genome and the application of DArT-seq to develop markers from a perennial wild relative of wheat, marking a paradigm shift from the seeming concentration of the technology on cultivated species. Integration of these markers with appropriate cytogenetic methods would accelerate development and characterization of wheat-alien chromosome introgression lines.

Quantifying fungal viability in air and water samples using quantitative PCR after treatment with propidium monoazide (PMA)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Vesper, Stephen; McKinstry, Craig A.; Hartmann, Chris

2007-11-28

A method is described to discriminate between live and dead cells of the infectious fungi Aspergillus fumigatus, Aspergillus flavus, Aspergillus terreus, Mucor racemosus, Rhizopus stolonifer and Paecilomyces variotii. To test the method, conidial suspensions were heat inactivated at 85 °C or held at 5 °C (controls) for 1 h. Polycarbonate filters (25 mm diameter, 0.8 μm pore size) were placed on "welled" slides (14 mm diameter) and the filters treated with either PBS or PMA. Propidium monoazide (PMA), which enters dead cells but not live cells, was incubated with cell suspensions, exposed to blue wavelength light-emitting diodes (LED) to inactivatemore » remaining PMA and secure intercalation of PMAwith DNA of dead cells. Treated cells were extracted and the live and dead cells evaluated with quantitative PCR (QPCR). After heat treatment and DNA modification with PMA, all fungal species tested showed an approximate 100- to 1000-fold difference in cell viability estimated by QPCR analysis which was consistent with estimates of viability based on culturing.« less

Fermentation process improvement of a Chinese traditional food: soybean residue cake.

PubMed

Yao, Yingzheng; Pan, Siyi; Wang, Kexing; Xu, Xiaoyun

2010-09-01

Fermentation process improvement of soybean residue cake, a Chinese traditional fermented food, and its physicochemical analysis during fermentation were studied. One of the dominant strains in the fermentation was isolated and identified as Mucor racemosus Fresenius. The fermentation process was improved by subsection fermentation. The crude protein content decreased from 19.95 ± 0.03% in the raw soybean residue to 16.85 ± 0.10% in the fermented products, and the formaldehyde nitrogen content increased from 0.068 ± 0.004% to 0.461 ± 0.022% in final fermented cakes. Hardness of samples significantly (P < 0.05) increased whereas springiness, cohesiveness, and resilience significantly (P < 0.05) decreased with increasing fermentation time, respectively. Microstructure observations showed obvious change of the surface of cake samples during the fermentation process. During the soybean processing, it will produce plenty of by-products, and the most part of them is soybean residue. The discarded soybean residue causes economic loss. Fortunately, we can obtain nutritious and delicious fermented soybean residue cakes by fermenting soybean residue as raw material.

Does co-inoculation of Lactuca serriola with endophytic and arbuscular mycorrhizal fungi improve plant growth in a polluted environment?

PubMed

Ważny, Rafał; Rozpądek, Piotr; Jędrzejczyk, Roman J; Śliwa, Marta; Stojakowska, Anna; Anielska, Teresa; Turnau, Katarzyna

2018-04-01

Phytoremediation of polluted sites can be improved by co-inoculation with mycorrhizal and endophytic fungi. In this study, the effects of single- and co-inoculation of Lactuca serriola with an arbuscular mycorrhizal (AM) fungus, Rhizoglomus intraradices, and endophytic fungi, Mucor sp. or Trichoderma asperellum, on plant growth, vitality, toxic metal accumulation, sesquiterpene lactone production and flavonoid concentration in the presence of toxic metals were evaluated. Inoculation with the AM fungus increased biomass yield of the plants grown on non-polluted and polluted substrate. Co-inoculation with the AM fungus and Mucor sp. resulted in increased biomass yield of plants cultivated on the polluted substrate, whereas co-inoculation with T. asperellum and the AM fungus increased plant biomass on the non-polluted substrate. In the presence of Mucor sp., mycorrhizal colonization and arbuscule richness were increased in the non-polluted substrate. Co-inoculation with the AM fungus and Mucor sp. increased Zn concentration in leaves and roots. The concentration of sesquiterpene lactones in plant leaves was decreased by AM fungus inoculation in both substrates. Despite enhanced host plant costs caused by maintaining symbiosis with numerous microorganisms, interaction of wild lettuce with both mycorrhizal and endophytic fungi was more beneficial than that with a single fungus. The study shows the potential of double inoculation in unfavourable environments, including agricultural areas and toxic metal-polluted areas.

Proteomics analysis of high lipid-producing strain Mucor circinelloides WJ11: an explanation for the mechanism of lipid accumulation at the proteomic level.

PubMed

Tang, Xin; Zan, Xinyi; Zhao, Lina; Chen, Haiqin; Chen, Yong Q; Chen, Wei; Song, Yuanda; Ratledge, Colin

2016-02-11

The oleaginous fungus, Mucor circinelloides, is attracting considerable interest as it produces oil rich in γ-linolenic acid. Nitrogen (N) deficiency is a common strategy to trigger the lipid accumulation in oleaginous microorganisms. Although a simple pathway from N depletion in the medium to lipid accumulation has been elucidated at the enzymatic level, global changes at protein levels upon N depletion have not been investigated. In this study, we have systematically analyzed the changes at the levels of protein expression in M. circinelloides WJ11, a high lipid-producing strain (36 %, lipid/cell dry weight), during lipid accumulation. Proteomic analysis demonstrated that N depletion increased the expression of glutamine synthetase, involved in ammonia assimilation, for the supply of cellular nitrogen but decreased the metabolism of amino acids. Upon N deficiency, many proteins (e.g., fructose-bisphosphate aldolase, glyceraldehyde-3-phosphate dehydrogenase, enolase, pyruvate kinase) involved in glycolytic pathway were up-regulated while proteins involved in the tricarboxylic acid cycle (e.g., isocitrate dehydrogenase, succinyl-CoA ligase, succinate dehydrogenase, fumarate hydratase) were down-regulated, indicating this activity was retarded thereby leading to a greater flux of carbon into fatty acid biosynthesis. Moreover, glucose-6-phosphate dehydrogenase, transaldolase and transketolase, which participate in the pentose phosphate pathway, were up-regulated, leading to the increased production of NADPH, the reducing power for fatty acid biosynthesis. Furthermore, protein and nucleic acid metabolism were down-regulated and some proteins involved in energy metabolism, signal transduction, molecular chaperone and redox homeostasis were up-regulated upon N depletion, which may be the cellular response to the stress produced by the onset of N deficiency. N limitation increased those expressions of the proteins involved in ammonia assimilation but decreased that

Antisecretory and antiulcer activity of Asparagus racemosus Willd. against indomethacin plus phyloric ligation-induced gastric ulcer in rats.

PubMed

Bhatnagar, Maheep; Sisodia, S S

2006-01-01

To study the antisecretory and antiulcer activity of Asparagus racemosus Willd. (methanolic extract) and its action against indomethacin (a non-steroidal anti-inflammatory drug) plus pyloric ligation (PL)-induced gastric ulcers in rats. Indomethacin plus PL-induced gastric ulceration model was used in the study. Treatment with Asparagus racemosus (Shatavari) crude extract (100 mg/kg/day orally) for fifteen days significantly reduced ulcer index when compared with control group. The reduction in gastric lesions was comparable to a standard antiulcer drug Ranitidine (30 mg/kg/ day orally). Crude extract also significantly reduced volume of gastric secretion, free acidity and total acidity. A significant increase in total carbohydrate (TC) and TC/total protein (TP) ratio of gastric juice was also observed. No significant change in the total protein was noted. Asparagus racemosus was found to be an effective antiulcerogenic agent, whose activity can well be compared with that of ranitidine hydrochloride. The results of this study suggest that Asparagus racemosus causes an inhibitory effect on release of gastric hydrochloric acid and protects gastric mucosal damage.

Evaluation of nested PCR in diagnosis of fungal rhinosinusitis.

PubMed

Badiee, Parisa; Gandomi, Behrooz; Sabz, Gholamabbass; Khodami, Bijan; Choopanizadeh, Maral; Jafarian, Hadis

2015-02-01

Given the importance of rapid diagnosis for fungal rhinosinusitis, this study aimed to evaluate the use of nested PCR to identify Aspergillus and Mucor species in clinical samples from patients with suspected fungal rhinosinusitis. Functional endoscopic sinus surgery specimens were collected from 98 patients with rhinosinusitis from 2012 to 2013. All samples were ground and cultured on sabouraud dextrose agar. The isolated fungi were identified based on their macroscopic and microscopic features. Fungal DNA was extracted from the tissue samples and nested PCR was performed with two sets of primers for Mucor and Aspergillus. Direct microscopic showed that 5.1% contained fungal components and 9.2% exhibited growth of fungi in culture. The most common agents isolated were Aspergillus fumigatus (n= 3), Aspergillus flavus (n=2), Penicillium sp (n=3) and Alternaria sp. (n=1). Mucor sp. was identified in the pathology smear from 1 patient. Positive results for fungal rhinosinusitis were obtained for a total of 10.2% by culture or pathology smear. Positive PCR results were obtained in 72 samples for Aspergillus and 31 samples for Mucor. Our results suggest that endoscopic sinus surgery specimens are not suitable for nested PCR, probably because of the accumulation of fungi that contaminate the environmental air. This drawback is a limiting factor for diagnosis with nasal cavity specimens. Therefore, molecular methods and conventional culture techniques are helpful complementary diagnostic methods to detect fungal rhinosinusitis and determine appropriate management for these patients.

Immunoaffinity Knockout of Saponin Glycosides from Asparagus racemosus to Assess Anti-lipid Peroxidation.

PubMed

Onlom, Churanya; Phrompittayarat, Watoo; Putalun, Waraporn; Waranuch, Neti; Ingkaninan, Kornkanok

2017-07-01

Asparagus racemosus Willd (Asparagaceae family), known as Shatavari, is important in Ayurveda and traditional Thai medicines. The saponin glycosides, shatavarin I and IV are major constituents in its roots and may be responsible for their actions including protection against lipid peroxidation and carcinogenesis. To develop an immunoaffinity column for isolating compounds with structures related to shatavarin IV from crude extracts of A. racemosus root. The monoclonal antibody recognising shatavarin IV (mAbShavIV) was coupled to an Affi-Gel Hz gel to isolate compounds with structures related to shatavarin IV from the other components of crude extracts of A. racemosus root. The saponin glycosides in each fraction were analysed by mAbShavIV ELISA and LC-MS/MS. The pooled wash-through fractions contained 3% of loaded mAbShavIV reactive saponin glycosides, while eluted fractions released ~ 90% of shatavarin saponin glycosides in a single step. Using thiobarbiturate (TBARs) to measure lipid-peroxidation, the extract, and the pooled wash-through fractions showed moderate protection against Cu + -induced oxidation of human low density lipoprotein (LDL) (IC 50 11.3 ± 1.4 and 12.6 ± 0.9 μg/mL, respectively). In contrast, the saponin glycosides eluted from the mAbShavIV-column had weaker protectant (IC 50 29.7 ± 1.8 μg/mL) suggesting that A. racemosus shatavarins do not inhibit carcinogenesis through preventing lipid peroxidation. The strategy described here demonstrates its utility for isolating a group of related compounds from the rest of the extract with selectivity and recovery rate. Pharmacological efficacy and synergistic effects of the components obtained can be further investigated. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

Cytogenetic and molecular identification of three Triticum aestivum-Leymus racemosus translocation addition lines.

PubMed

Wang, Le; Yuan, Jianhua; Bie, Tongde; Zhou, Bo; Chen, Peidu

2009-06-01

Chromosome 2C from Aegilops cylindrica has the ability to induce chromosome breakage in common wheat (Tritivum aestivum). In the BC(1)F(3) generation of the T. aestivum cv. Chinese Spring and a hybrid between T. aestivum-Leymus racemosus Lr.7 addition line and T. aestivum-Ae. cylindrica 2C addition line, three disomic translocation addition lines (2n = 44) were selected by mitotic chromosome C-banding and genomic in situ hybridization. We further characterized these T. aestivum-L. racemosus translocation addition lines, NAU636, NAU637 and NAU638, by chromosome C-banding, in situ hybridization using the A- and D-genome-specific bacterial artificial chromosome (BAC) clones 676D4 and 9M13; plasmids pAs1 and pSc119.2, and 45S rDNA; as well as genomic DNA of L. racemosus as probes, in combination with double ditelosomic test cross and SSR marker analysis. The translocation chromosomes were designated as T3AS-Lr7S, T6BS-Lr7S, and T5DS-Lr7L. The translocation line T3AS-Lr7S was highly resistant to Fusarium head blight and will be useful germplasm for resistance breeding.

Effect of Novel Starter Culture on Reduction of Biogenic Amines, Quality Improvement, and Sensory Properties of Doenjang, a Traditional Korean Soybean Fermented Sauce Variety.

PubMed

Shukla, Shruti; Lee, Jong Suk; Park, Hae-Kyong; Yoo, Jung-Ah; Hong, Sung-Yong; Kim, Jong-Kyu; Kim, Myunghee

2015-08-01

To select appropriate microorganisms as starter cultures for the reliable and reproducible fermentation of soybean fermented products of Korean Doenjang, various ratios of fungi (Aspergillus oryzae J, Mucor racemosus 15, M. racemosus 42) combined with Bacillus subtilis TKSP 24 were selected as either single, double, or multiple Meju strains for commercial mass production of Doenjang, followed by analysis of sensory characteristics. In the sensory evaluation, Doenjang BAM15-1 and BAM42-1, which were fermented with multiple strains (1:1:1), showed the highest sensory scores as compared to control. Based on sensory characteristics, 6 Doenjang samples were subjected to quantitative determination of amino acids, free sugars, and organic acids (volatile and nonvolatile) contents, followed by determination of biogenic amines. Total sweet taste amino acid contents were highest in BAM15-1 and BAM42-1 samples (333.7 and 295.8 mg/100 g, respectively) and similar that of control (391.1 mg/100 g). Samples BAM15-1 and BAM42-1 showed the relatively high volatile and nonvolatile organic acid contents (154.24, 192.26, and 71.31, 82.42 mg/100 g, respectively). In addition, BAM15-1 and BAM42-1 showed negligible biogenic amine formation, ranging from 0.00 to 1.02 and 0.00 to 3.92 mg/100 g, respectively. These findings indicate that determination of food components along with sensory and quality attributes using multiple microbial Meju strains as a starter culture may provide substantial results on improved quality fermented Doenjang products. © 2015 Institute of Food Technologists®

Microbial burden of some herbal antimalarials marketed at Elele, Rivers State.

PubMed

Tatfeng, Y M; Olama, E H; Ojo, T O

2009-12-30

Herbal antimalarials still remain an alternative to our traditional communities who can not afford orthodox antimalarials. This study was aimed at investigating the microbial quality of six herbal antimalarials using standard microbiological methods. Of the six preparations analyzed, "schnapps", palm wine and water were the media of preparation; the water base preparations recorded higher microbial load. The mean microbial load was 159.5 × 10(5) cfu/ml and 217.4 × 10(2)cfu/ml in water and alcohol base preparations respectively. The microbial profile of the preparations showed that the schnapps base preparations were predominantly contaminated with Bacillus sp (Aerobic spore bearers) and Mucor spp. The palm wine preparation harboured Bacillus sp, yeasts and Mucor spp while the water base preparations had several isolates such as Staphylococcus epidermidis, Pseudomonas aeruginosa, Escherichia coli 0157H7, Proteus mirabilis, Enterococcus feacalis, Serratia marcensces, Staph. aureus, Bacillus spp and Mucor spp. Conclusively, this study underlines the public health importance of these preparations given the high burden of such human pathogen as Ecoli O157H7, Ps aeruginosa, Stahp aureus, etc. in the preparations.

The Mycobiota of Air Inside and Outside the Meju Fermentation Room and the Origin of Meju Fungi.

PubMed

Kim, Dae-Ho; Kim, Sun-Hwa; Kwon, Soon-Wo; Lee, Jong-Kyu; Hong, Seung-Beom

2015-09-01

The fungi on Meju are known to play an important role as degrader of macromolecule of soybeans. In order to elucidate the origin of fungi on traditional Meju, mycobiota of the air both inside and outside traditional Meju fermentation rooms was examined. From 11 samples of air collected from inside and outside of 7 Meju fermentation rooms, 37 genera and 90 species of fungi were identified. In outside air of the fermentation room, Cladosporium sp. and Cladosporium cladosporioides were the dominant species, followed by Cladosporium tenuissimum, Eurotium sp., Phoma sp., Sistotrema brinkmannii, Alternaria sp., Aspergillus fumigatus, Schizophyllum commune, and Penicillium glabrum. In inside air of the fermentation room, Cladosporium sp., Aspergillus oryzae, Penicillium chrysogenum, Asp. nidulans, Aspergillus sp., Cla. cladosporioides, Eurotium sp., Penicillium sp., Cla. tenuissimum, Asp. niger, Eur. herbariorum, Asp. sydowii, and Eur. repens were collected with high frequency. The concentrations of the genera Aspergillus, Eurotium, and Penicillium were significantly higher in inside air than outside air. From this result and those of previous reports, the origin of fungi present on Meju was inferred. Of the dominant fungal species present on Meju, Lichtheimia ramosa, Mucor circinelloides, Mucor racemosus, and Scopulariopsis brevicaulis are thought to be originated from outside air, because these species are not or are rarely isolated from rice straw and soybean; however, they were detected outside air of fermentation room and are species commonly found in indoor environments. However, Asp. oryzae, Pen. polonicum, Eur. repens, Pen. solitum, and Eur. chevalieri, which are frequently found on Meju, are common in rice straw and could be transferred from rice straw to Meju. The fungi grow and produce abundant spores during Meju fermentation, and after the spores accumulate in the air of fermentation room, they could influence mycobiota of Meju fermentation in the following

The Mycobiota of Air Inside and Outside the Meju Fermentation Room and the Origin of Meju Fungi

PubMed Central

Kim, Dae-Ho; Kim, Sun-Hwa; Kwon, Soon-wo; Lee, Jong-Kyu

2015-01-01

The fungi on Meju are known to play an important role as degrader of macromolecule of soybeans. In order to elucidate the origin of fungi on traditional Meju, mycobiota of the air both inside and outside traditional Meju fermentation rooms was examined. From 11 samples of air collected from inside and outside of 7 Meju fermentation rooms, 37 genera and 90 species of fungi were identified. In outside air of the fermentation room, Cladosporium sp. and Cladosporium cladosporioides were the dominant species, followed by Cladosporium tenuissimum, Eurotium sp., Phoma sp., Sistotrema brinkmannii, Alternaria sp., Aspergillus fumigatus, Schizophyllum commune, and Penicillium glabrum. In inside air of the fermentation room, Cladosporium sp., Aspergillus oryzae, Penicillium chrysogenum, Asp. nidulans, Aspergillus sp., Cla. cladosporioides, Eurotium sp., Penicillium sp., Cla. tenuissimum, Asp. niger, Eur. herbariorum, Asp. sydowii, and Eur. repens were collected with high frequency. The concentrations of the genera Aspergillus, Eurotium, and Penicillium were significantly higher in inside air than outside air. From this result and those of previous reports, the origin of fungi present on Meju was inferred. Of the dominant fungal species present on Meju, Lichtheimia ramosa, Mucor circinelloides, Mucor racemosus, and Scopulariopsis brevicaulis are thought to be originated from outside air, because these species are not or are rarely isolated from rice straw and soybean; however, they were detected outside air of fermentation room and are species commonly found in indoor environments. However, Asp. oryzae, Pen. polonicum, Eur. repens, Pen. solitum, and Eur. chevalieri, which are frequently found on Meju, are common in rice straw and could be transferred from rice straw to Meju. The fungi grow and produce abundant spores during Meju fermentation, and after the spores accumulate in the air of fermentation room, they could influence mycobiota of Meju fermentation in the following

Outbreak of Invasive Wound Mucormycosis in a Burn Unit Due to Multiple Strains of Mucor circinelloides f. circinelloides Resolved by Whole-Genome Sequencing.

PubMed

Garcia-Hermoso, Dea; Criscuolo, Alexis; Lee, Soo Chan; Legrand, Matthieu; Chaouat, Marc; Denis, Blandine; Lafaurie, Matthieu; Rouveau, Martine; Soler, Charles; Schaal, Jean-Vivien; Mimoun, Maurice; Mebazaa, Alexandre; Heitman, Joseph; Dromer, Françoise; Brisse, Sylvain; Bretagne, Stéphane; Alanio, Alexandre

2018-04-24

Mucorales are ubiquitous environmental molds responsible for mucormycosis in diabetic, immunocompromised, and severely burned patients. Small outbreaks of invasive wound mucormycosis (IWM) have already been reported in burn units without extensive microbiological investigations. We faced an outbreak of IWM in our center and investigated the clinical isolates with whole-genome sequencing (WGS) analysis. We analyzed M. circinelloides isolates from patients in our burn unit (BU1, Hôpital Saint-Louis, Paris, France) together with nonoutbreak isolates from Burn Unit 2 (BU2, Paris area) and from France over a 2-year period (2013 to 2015). A total of 21 isolates, including 14 isolates from six BU1 patients, were analyzed by whole-genome sequencing (WGS). Phylogenetic classification based on de novo assembly and assembly free approaches showed that the clinical isolates clustered in four highly divergent clades. Clade 1 contained at least one of the strains from the six epidemiologically linked BU1 patients. The clinical isolates were specific to each patient. Two patients were infected with more than two strains from different clades, suggesting that an environmental reservoir of clonally unrelated isolates was the source of contamination. Only two patients from BU1 shared one strain, which could correspond to direct transmission or contamination with the same environmental source. In conclusion, WGS of several isolates per patients coupled with precise epidemiological data revealed a complex situation combining potential cross-transmission between patients and multiple contaminations with a heterogeneous pool of strains from a cryptic environmental reservoir. IMPORTANCE Invasive wound mucormycosis (IWM) is a severe infection due to environmental molds belonging to the order Mucorales. Severely burned patients are particularly at risk for IWM. Here, we used whole-genome sequencing (WGS) analysis to resolve an outbreak of IWM due to Mucor circinelloides that occurred in

"Asparagus Racemosus" Enhances Memory and Protects against Amnesia in Rodent Models

ERIC Educational Resources Information Center

Ojha, Rakesh; Sahu, Alakh N.; Muruganandam, A. V.; Singh, Gireesh Kumar; Krishnamurthy, Sairam

2010-01-01

"Asparagus Racemosus" (AR) is an Ayurvedic rasayana possessing multiple neuropharmacological activities. The adpatogenic and antidepressant activity of AR is well documented. The present study was undertaken to assess nootropic and anti-amnesic activities of MAR in rats. The Morris water maze (MWM) and elevated plus maze (EPM) models were employed…

Selective effects of two systemic fungicides on soil fungi.

PubMed

Abdel-Fattah, H M; Abdel-Kader, M I; Hamida, S

1982-08-20

BAS 317 00F was not toxic to the total count of fungi after 2 days but was regularly significantly toxic at the three doses after 5, 20 and 40 days and toxic at the low and the high doses after 80 days. In the agar medium, it was toxic to the counts of total fungi. Aspergillus, A. terreus, Rhizopus oryzae and Mucor racemosus at the high dose. Only the mycelial growth of Trichoderma viride which was significantly inhibited by the three doses when this fungicide was added to the liquid medium. Polyram-Combi induced two effects on the total population of soil fungi. One inhibitory and this was demonstrated almost regularly after 2, 10 and 40 days and the other stimulatory after 80 days of treatment with the low and the high doses. In the agar medium, this fungicide was very toxic to total fungi and to almost all fungal genera and species at the three doses. Several fungi could survive the high dose. In liquid medium, the test fungi showed variable degree of sensitivity and the most sensitive was Gliocladium roseum which was completely eradicated by the three doses.

Detection of fungi colony growth on bones by dynamic speckle

NASA Astrophysics Data System (ADS)

Vincitorio, F. M.; Budini, N.; Mulone, C.; Spector, M.; Freyre, C.; López Díaz, A. J.; Ramil, A.

2013-11-01

In this work we have studied the dynamic speckle patterns of mucor fungi colonies, which were inoculated on different samples. We were interested in analyzing the development of fungi colonies in bones, since during the last two years, a series of infections by mucor fungi have been reported on patients from different hospitals in Argentina. Coincidentally, all of these infections appeared on patients that were subjected to a surgical intervention for implantation of a titanium prosthesis. Apparently, the reason of the infection was a deficient sterilization process in conjunction with an accidental contamination. We observed that fungi growth, activity and death can be distinguished by means of the dynamic speckle technique.

Longterm storage of post-packaged bread by controlling spoilage pathogens using Lactobacillus fermentum C14 isolated from homemade curd

PubMed Central

Barman, Soma; Ghosh, Ranjan; Sengupta, Shreya

2017-01-01

One potent lactic acid bacterial strain C14 with strong antifungal activity was isolated from homemade curd. Based on morphological as well as biochemical characters and 16S rDNA sequence homology the strain was identified as Lactobacillus fermentum. It displayed a wide antimicrobial spectrum against both Gram-positive and Gram-negative pathogenic bacteria, and also against number of food spoilage, plant and human pathogenic fungi. The cell free supernatant (CFS) of the strain C14 was also effective against the fungi tested. Inhibition of radial growth of Penicillium digitatum, Trichophyton rubrum and Mucor sp. was noticed in the presence of CFS of C14 even at low concentration (1%). More than 94.3 ± 1.6% and 91.5 ± 2.2% inhibition of conidial germination of P. digitatum and Mucor sp. were noticed in the presence of 10-fold-concentrated CFS of C14. Massive deformation of the fungal mycelia was observed by SEM studies, and losses of cellular proteins and DNA are also evident upon its treatment with C14. HPLC analysis revealed the presence of phenyl lactic acid, lactic acid along with some unidentified compounds in the antifungal extract. Challenge experiment showed immense potential of the strain C14 in preventing the spoilage of bread samples caused by Mucor sp. and Bacillus subtilis. The bread samples remained fresh upto 25 days even after inoculation with Mucor sp. (3.7 × 104 spores /ml) and B. subtilis (4.6 × 104 CFU /ml). Along with the antifungal properties, the isolated lactic acid bacterial strain also showed very good antioxidant activities. Unchanged level of liver enzymes serum glutamic pyruvic transaminase and serum glutamic oxaloacetic transaminase in albino mice upon feeding with C14 also suggested non-toxic nature of the bacterial isolate. PMID:28859156

Longterm storage of post-packaged bread by controlling spoilage pathogens using Lactobacillus fermentum C14 isolated from homemade curd.

PubMed

Barman, Soma; Ghosh, Ranjan; Sengupta, Shreya; Mandal, Narayan C

2017-01-01

One potent lactic acid bacterial strain C14 with strong antifungal activity was isolated from homemade curd. Based on morphological as well as biochemical characters and 16S rDNA sequence homology the strain was identified as Lactobacillus fermentum. It displayed a wide antimicrobial spectrum against both Gram-positive and Gram-negative pathogenic bacteria, and also against number of food spoilage, plant and human pathogenic fungi. The cell free supernatant (CFS) of the strain C14 was also effective against the fungi tested. Inhibition of radial growth of Penicillium digitatum, Trichophyton rubrum and Mucor sp. was noticed in the presence of CFS of C14 even at low concentration (1%). More than 94.3 ± 1.6% and 91.5 ± 2.2% inhibition of conidial germination of P. digitatum and Mucor sp. were noticed in the presence of 10-fold-concentrated CFS of C14. Massive deformation of the fungal mycelia was observed by SEM studies, and losses of cellular proteins and DNA are also evident upon its treatment with C14. HPLC analysis revealed the presence of phenyl lactic acid, lactic acid along with some unidentified compounds in the antifungal extract. Challenge experiment showed immense potential of the strain C14 in preventing the spoilage of bread samples caused by Mucor sp. and Bacillus subtilis. The bread samples remained fresh upto 25 days even after inoculation with Mucor sp. (3.7 × 104 spores /ml) and B. subtilis (4.6 × 104 CFU /ml). Along with the antifungal properties, the isolated lactic acid bacterial strain also showed very good antioxidant activities. Unchanged level of liver enzymes serum glutamic pyruvic transaminase and serum glutamic oxaloacetic transaminase in albino mice upon feeding with C14 also suggested non-toxic nature of the bacterial isolate.

Identification of fungal ene-reductase activity by means of a functional screening.

PubMed

Romagnolo, Alice; Spina, Federica; Brenna, Elisabetta; Crotti, Michele; Parmeggiani, Fabio; Varese, Giovanna Cristina

2015-06-01

Bioeconomy stresses the need of green processes promoting the development of new methods for biocatalyzed alkene reductions. A functional screening of 28 fungi belonging to Ascomycota, Basidiomycota, and Zygomycota isolated from different habitats was performed to analyze their capability to reduce C=C double bonds towards three substrates (cyclohexenone, α-methylnitrostyrene, and α-methylcinnamaldehyde) with different electron-withdrawing groups, i.e., ketone, nitro, and aldehyde, respectively. Almost all the fungi showed this reducing activity. Noteworthy Gliomastix masseei, Mucor circinelloides, and Mucor plumbeus resulted versatile and effective, being able to reduce all the model substrates quickly and with high yields. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Cytological effects of fungal metabolites produced by fungi isolated from Egyptian poultry feedstuffs.

PubMed

Abdou, R F; Megalla, S E; Moharram, A M; Abdel-Gawad, K M; Sherif, T H; el-Syed Mahmood, A L; Lottfy, A E

1989-01-01

The cytogenetic effects of fungal metabolites produced by 113 strains belonging to 36 fungal species and isolated form 5 substrates of commercial poultry feedstuffs were tested for their effect on the growing root meristems of Allium cepa. The fungal metabolites of Paecilomyces canescens, Aspergillus fumigatus, Syncephalastrum racemosum, Aspergillus terreus and Mucor hiemalis strongly suppressed cell division. Metabolites from other strains had less effect on cell division but permitted the appearance of several abnormalities through different mitotic stages. In general, chromosomal aberrations were more obvious with metabolites of Aspergillus species, Mucor circinelloides and Cladosporium cladosporioides. The mutagenic effects produced by these fungal metabolites reflect the risk that might take place through the consumption of these contaminated feedstuffs.

Identification of Mucorales isolates from soil using morphological and molecular methods.

PubMed

Ziaee, A; Zia, M; Bayat, M; Hashemi, J

2016-03-01

Soil is the main habitat of saprophytic and pathogenic fungi. Mucoromycotina constitutes a large group of soil fungi, with certain opportunistic members causing systemic infections in immunocompromised hosts. The majority of human and animal infections are caused by the members of the genera Rhizopus , Mucor , Rhizomucor , Lichtheimia ( Absidia) , Cunninghamella, and Mortierella . Accordingly, in the present study, we aimed to isolate and identify the main genera of the order Mucorales, using molecular assays and morphological features . In total, 340 soil samples were collected from seven public parks throughout the city and sidewalk gardens in 14 municipal districts in Isfahan, Iran. All the samples were cultured on the appropriate media, incubated at 27°C for 2- 4 days, and examined daily for visible fungal growth. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was applied and macroscopic, microscopic, and physiological characteristics were assessed to identify fungal colonies. 400 pure colonies, belonging to the orders Mucorales and Mortierellales, including the genera Lichtheimia , Rhizopus, Rhizomucor, Mucor, Cunninghamella, and Mortierella, were identified . The genus Rhizopus (35.5%) was the most frequent isolate, followed by Mucor (32.25%) and Rhizomucor (27.5%). The results emphasize the importance of opportunistic fungi in public areas and indicate the risk of exposure for immunocompromised individuals.

Biodegradation of dieldrin by a soil fungus isolated from a soil with annual endosulfan applications.

PubMed

Kataoka, Ryota; Takagi, Kazuhiro; Kamei, Ichiro; Kiyota, Hiromasa; Sato, Yuuki

2010-08-15

An aerobic dieldrin-degrading fungus, Mucor racemosus strain DDF, was isolated from a soil to which endosulfan had been annually applied for more than 10 years until 2008. Strain DDF degraded dieldrin to 1.01 microM from 14.3 microM during a 10-day incubation at 25 degrees C. Approximately 0.15 microM (9%) of aldrin trans-diol was generated from the dieldrin degradation after a 1-day incubation. The degradation of dieldrin by strain DDF was detected over a broad range of pH and concentrations of glucose and nitrogen sources. Extracellular fluid without mycelia also degraded dieldrin. Strain DDF degraded not only dieldrin but also heptachlor, heptachlor epoxide, endosulfan, endosulfan sulfate, DDT, and DDE. Endosulfan sulfate and heptachlor were degraded by 0.64 microM (95%) and 0.75 microM (94%), respectively, whereas endosulfan and DDE were degraded by 2.42 microM (80%) and 3.29 microM (79%), respectively, and DDT and heptachlor epoxide were degraded by 6.95 microM (49.3%) and 5.36 microM (67.5%), respectively, compared with the control, which had a concentration of approximately 14 microM. These results suggest that strain DDF could be a candidate for the bioremediation of sites contaminated with various persistent organochlorine pesticides including POPs.

Fungal biodegradation of anthracene-polluted cork: A comparative study.

PubMed

Jové, Patrícia; Olivella, Maria À; Camarero, Susana; Caixach, Josep; Planas, Carles; Cano, Laura; De Las Heras, Francesc X

2016-01-01

The efficiency of cork waste in adsorbing aqueous polycyclic aromatic hydrocarbons (PAHs) has been previously reported. Biodegradation of contaminated cork using filamentous fungi could be a good alternative for detoxifying cork to facilitate its final processing. For this purpose, the degradation efficiency of anthracene by three ligninolytic white-rot fungi (Phanerochaete chrysosporium, Irpex lacteus and Pleurotus ostreatus) and three non-ligninolytic fungi which are found in the cork itself (Aspergillus niger, Penicillium simplicissimum and Mucor racemosus) are compared. Anthracene degradation by all fungi was examined in solid-phase cultures after 0, 16, 30 and 61 days. The degradation products of anthracene by P. simplicissimum and I. lacteus were also identified by GC-MS and a metabolic pathway was proposed for P. simplicissimum. Results show that all the fungi tested degraded anthracene. After 61 days of incubation, approximately 86%, 40%, and 38% of the initial concentration of anthracene (i.e., 100 µM) was degraded by P. simplicissimum, P. chrysosporium and I. lacteus, respectively. The rest of the fungi degraded anthracene to a lesser extent (<30%). As a final remark, the results obtained in this study indicate that P. simplicissimum, a non-ligninolytic fungi characteristic of cork itself, could be used as an efficient degrader of PAH-contaminated cork.

Invasive Mucormycosis Induced Pneumopericardium: A Rare Cause of Pneumopericardium in an Immunocompromised Patient

PubMed Central

Khan, Sana; Waqar Elahi, Muhammad; Ullah, Waqas; Ahmad, Ejaz; Al Mohajer, Mayar; Majeed, Aneela

2017-01-01

Mucor and Rhizopus cause life-threatening infections primarily involving the lungs and sinuses, which disseminate very rapidly by necrosis and infarction of the contiguous tissues. We present a case of a 64-year-old African American posttransplant patient who presented with a productive cough and weight loss. He had a past surgical history of renal transplant for renal cell carcinoma and was on dual immunosuppressive therapy, that is, mycophenolate and tacrolimus. During his hospital stay, he developed a pneumopericardium due to the direct extension of a lung lesion. The diagnosis was made by radiological imaging and PCR result which was consistent with Mucor species. He was treated with antifungal therapy. The purpose of this report is to highlight the unusual association of mucormycosis with pneumopericardium. PMID:28596926

Identification of Mucorales isolates from soil using morphological and molecular methods

PubMed Central

Ziaee, A; Zia, M; Bayat, M; Hashemi, J

2016-01-01

Background and Purpose: Soil is the main habitat of saprophytic and pathogenic fungi. Mucoromycotina constitutes a large group of soil fungi, with certain opportunistic members causing systemic infections in immunocompromised hosts. The majority of human and animal infections are caused by the members of the genera Rhizopus, Mucor, Rhizomucor, Lichtheimia (Absidia), Cunninghamella, and Mortierella. Accordingly, in the present study, we aimed to isolate and identify the main genera of the order Mucorales, using molecular assays and morphological features. Materials and Methods: In total, 340 soil samples were collected from seven public parks throughout the city and sidewalk gardens in 14 municipal districts in Isfahan, Iran. All the samples were cultured on the appropriate media, incubated at 27°C for 2- 4 days, and examined daily for visible fungal growth. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was applied and macroscopic, microscopic, and physiological characteristics were assessed to identify fungal colonies. Results: 400 pure colonies, belonging to the orders Mucorales and Mortierellales, including the genera Lichtheimia, Rhizopus, Rhizomucor, Mucor, Cunninghamella, and Mortierella, were identified. The genus Rhizopus (35.5%) was the most frequent isolate, followed by Mucor (32.25%) and Rhizomucor (27.5%). Conclusion: The results emphasize the importance of opportunistic fungi in public areas and indicate the risk of exposure for immunocompromised individuals. PMID:28681007

Coprophilous Mucorales (ex Zygomycota) from three areas in the semi-arid of Pernambuco, Brazil.

PubMed

Souza, Carlos Alberto Fragoso de; Lima, Diogo Xavier; Gurgel, Luciana M S; Santiago, André Luiz Cabral Monteiro de Azevedo

Mucorales comprises fungi commonly isolated as saprobes from soil, dung, stored grains and plants. Although these fungi have been studied in several countries, there are relatively a few reports of them in semi-arid areas. Therefore, the aims of the present study were to assess and compare the Mucorales communities in dung from different species and breeds of herbivores in the semi-arid of Pernambuco, based on the frequency of occurrence and species richness of these fungi. Samples of dung collected in the cities of Arcoverde, Serra Talhada and Sertânia were incubated in moist chambers in triplicate. Altogether, 24 taxa of Mucorales distributed in the genera Absidia, Circinella, Cunninghamella, Lichtheimia, Mucor, Pilobolus, Rhizopus and Syncephalastrum were identified. The highest species richness was found in sheep excrement. Mucor circinelloides f. griseo-cyanus was the most common taxon, followed by M. ramosissimus. The similarity of the composition of Mucorales species was greatest between the excrements of Guzerá and Sindi breeds (bovine). All mucoralean species isolated are being cited for the first time from animal dung found in Caatinga and a new species of Mucor was recorded. An identification key for species of Mucorales from dung in the semi-arid region of Brazil is provided. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Rapid screening for human-pathogenic Mucorales using rolling circle amplification.

PubMed

Dolatabadi, S; Najafzadeh, M J; de Hoog, G S

2014-12-01

Mucormycosis has emerged as a relatively common severe mycosis in patients with haematological and allogeneic stem cell transplantation. Source of transmission is from unidentified sources in the environment. Early diagnosis of infection and its source of contamination are paramount for rapid and appropriate therapy. In this study, rolling circle amplification (RCA) is introduced as a sensitive, specific and reproducible isothermal DNA amplification technique for rapid molecular identification of six of the most virulent species (Rhizopus microsporus, R. arrhizus var. arrhizus, R. arrhizus var. delemar, Mucor irregularis, Mucor circinelloides, Lichtheimia ramosa, Lichtheimia corymbifera). DNAs of target species were successfully amplified, with no cross reactivity between species. RCA can be considered as a rapid detection method with high specificity and sensitivity, suitable for large screening. © 2014 Blackwell Verlag GmbH.

Use of external ventriculostomy and intrathecal anti-fungal treatment in cerebral mucormycotic abscess.

PubMed

Grannan, Benjamin L; Yanamadala, Vijay; Venteicher, Andrew S; Walcott, Brian P; Barr, John C

2014-10-01

Mucormycosis is an invasive fungal infection associated with a high mortality. Cerebral mucor abscesses can result secondary to rhinocerebral or hematogenous spread. Amphotericin B, posaconazole, and aggressive surgical resection are the hallmarks of treatment. While amphotericin is typically administered intravenously, less is known about the use of intrathecal amphotericin B. We describe a 42-year-old man who developed a cerebellar mucor abscess after undergoing hematopoietic stem cell transplant for the treatment of myelodysplastic syndrome. In the post-operative period he was admitted to the neurocritical care unit and received liposomal amphotericin B intravenously and through an external ventricular drain. This patient demonstrates that utilization of an external ventricular drain for intrathecal antifungal therapy in the post-operative period may warrant further study in patients with difficult to treat intracranial fungal abscesses. Copyright © 2014. Published by Elsevier Ltd.

Molecular and morphological identification of fungal species isolated from bealmijang meju.

PubMed

Kim, Ji Yeun; Yeo, Soo-Hwan; Baek, Sung Yeol; Choi, Hye Sun

2011-12-01

Bealmijang is a short-term aged paste made from meju, which is a brick of fermented soybeans and other ingredients. Different types of bealmijang are available depending on the geographic region or ingredients used. However, no study has clarified the microbial diversity of these types. We identified 17 and 14 fungal species from black soybean meju (BSM) and buckwheat meju (BWM), respectively, on the basis of morphology, culture characteristics, and internal transcribed spacer and beta-tubulin gene sequencing. In both meju, Aspergillus oryzae, Rhizopus oryzae, Penicillium polonicum, P. steckii, Cladosporium tenuissimum, C. cladosporioides, C. uredinicola, and yeast species Pichia burtonii were commonly found. Moreover, A. flavus, A. niger, P. crustosum, P. citrinum, Eurotium niveoglaucum, Absidia corymbifera, Setomelanomma holmii, Cladosporium spp. and unclassified species were identified from BSM. A. clavatus, Mucor circinelloides, M. racemosus, P. brevicompactum, Davidiella tassiana, and Cladosporium spp. were isolated from BWM. Fast growing Zygomycetous fungi is considered important for the early stage of meju fermentation, and A. oryae and A. niger might play a pivotal role in meju fermentation owing to their excellent enzyme productive activities. It is supposed that Penicillium sp. and Pichia burtonii could contribute to the flavor of the final food products. Identification of this fungal diversity will be useful for understanding the microbiota that participate in meju fermentation, and these fungal isolates can be utilized in the fermented foods and biotechnology industries.

Phyllanthus wightianus Müll. Arg.: A Potential Source for Natural Antimicrobial Agents

PubMed Central

Natarajan, D.; Srinivasan, R.; Shivakumar, M. S.

2014-01-01

Phyllanthus wightianus belongs to Euphorbiaceae family having ethnobotanical importance. The present study deals with validating the antimicrobial potential of solvent leaf extracts of P. wightianus. 11 human bacterial pathogens (Bacillus subtilis, Streptococcus pneumoniae, Staphylococcus epidermidis, Proteus vulgaris, Pseudomonas aeruginosa, Klebsiella pneumoniae, Salmonella typhimurium, Escherichia coli, Shigella flexneri, Proteus vulgaris, and Serratia marcescens) and 4 fungal pathogens (Candida albicans, Cryptococcus neoformans, Mucor racemosus, and Aspergillus niger) were also challenged with solvent leaf extracts usingagar well and disc diffusion methods. Further, identification of the active component present in the bioactive extract was done using GC-MS analysis. Results show that all extracts exhibited broad spectrum (6–29 mm) of antibacterial activity on most of the tested organisms. The results highlight the fact that the well in agar method was more effective than disc diffusion method. Significant antimicrobial activity was detected in methanol extract against S. pneumoniae (29 mm) with MIC and MBC values of 15.62 μg/mL. GC-MS analysis revealed that 29 bioactive constituents were present in methanolic extract of P. wightianus, of which 9,12-octadecaenioic acid (peak area 22.82%; RT-23.97) and N-hexadecanoic acid (peak area 21.55% RT-21.796) are the major compounds. The findings of this study show that P. wightianus extracts may be used as an anti-infective agent in folklore medicine. PMID:24883301

Integrated genomics of Mucorales reveals novel therapeutic targets

USDA-ARS?s Scientific Manuscript database

Mucormycosis is a life-threatening infection caused by Mucorales fungi. We sequenced 30 fungal genomes and performed transcriptomics with three representative Rhizopus and Mucor strains with human airway epithelial cells during fungal invasion to reveal key host and fungal determinants contributing ...

Microbial metabolism part 13 metabolites of hesperetin

USDA-ARS?s Scientific Manuscript database

The fungal culture, Mucor ramannianus (ATCC 2628) transformed hesperitin to four metabolites: 4'-methoxy -5, 7, 8, 3'-tetrahydroxyflavanone (8-hydroxyhesperetin), 5, 7, 3', 4'-tetrahydroxyflavanone (eriodictyol), 4'-methoxy-5, 3'-dihydroxyflavanone 7-sulfate (hesperetin 7-sulfate) and 5, 7, 3'-tri...

Fungi from interior organs of free-living small mammals in Czechoslovakia and Yugoslavia.

PubMed

Hubálek, Z; Rosický, B; Otcenásek, M

1980-01-01

A total of 308 fungi was isolated from interior organs (lungs, spleen, liver) of 529 small mammals belonging to 21 species, 7 families and 3 orders (Insectivora, Chiroptera, Rodentia), some of these being potentially pathogenic to vertebrates (e.g. Aspergillus flavus, A. fumigatus, Geotrichum candidum, Mucor pusillus, Rhizopus arrhizus). In one vole (Microtus arvalis) captured in South Moravia, adiaspiromycosis (Emmonsia crescens) was demonstrated. Comparison of mycoflora of hair and that of interior organs of wild small mammals revealed that out of the total number of isolates the following fungi were represented in a higher proportion from visceral organs than from the hair: Aspergillus (A. amstelodami, A. flavus, A. repens), Aureobasidium (A. pullulans), Candida, Cladosporium (C. herbarum), Cryptococcus, Fusarium, Gliocladium (G. deliquescens), Helminthosporium, Kloeckera, Mucor (M. fragilis, M. hiemalis, M. pusillus), Paecilomyces marquandii, Penicillium (P. purpurogenum), Phoma, Rhizopus arrhizus, Scopulariopsis (S. candida, S. koningii) and Torulopsis.

Microbial Metabolism. Part 10. Metabolites of 7,8 Dimethoxyflavone and 5-Methoxyflavone

USDA-ARS?s Scientific Manuscript database

Microbial transformation of 7, 8-dimethoxyflavone (1) by Mucor ramannianus (ATCC 9628) produced five metabolites: 7, 8-dimethoxy-4'-hydroxyflavone (2), 3', 4'-dihydroxy-7, 8-dimethoxyflavone (3), 7, 3'-dihydroxy-8-methoxyflavone (4), 7, 4'-dihydroxy-8-methoxyflavone (5) and 8-methoxy-7, 3', 4'-trihy...

Using homogenization, sonication and thermo-sonication to inactivate fungi

PubMed Central

Bevilacqua, Antonio; Sinigaglia, Milena; Corbo, Maria Rosaria

2016-01-01

Ultrasound (US), Thermo-sonication (TS) and High Pressure Homogenization (HPH) were studied as tools to inactivate the spores of Penicillium spp. and Mucor spp. inoculated in distilled water. For US, the power ranged from 40% to 100%, pulse from 2 to 10 s, and duration of the treatment from 2 to 10 min. TS was performed combining US (40–80% of power, for 8 min and pulse of 2 s) with a thermal treatment (50, 55 and 60°C at 4, 8 and 12 min). Homogenization was done at 30–150 MPa for 1, 2 and 3 times. Power was the most important factors to determine the antifungal effect of US and TS towards the conidia of Penicillium spp.; on the other hand, in US treatments Mucor spp. was also affected by pulse and time. HPH exerted a significant antifungal effect only if the highest pressures were applied for 2–3 times. PMID:27375964

Contribution of selected fungi to the reduction of cyanogen levels during solid substrate fermentation of cassava.

PubMed

Essers, A J; Jurgens, C M; Nout, M J

1995-07-01

The effect of six individual strains of the dominant microflora in solid substrate fermenting cassava on cyanogen levels was examined. Six out of eight batches of disinfected cassava root pieces were incubated for 72 h after inoculation with either of the fungi Geotrichum candidum, Mucor racemosus, Neurospora sitophila, Rhizopus oryzae and Rhizopus stolonifer, or a Bacillus sp., isolated from on-farm fermented cassava flours from Uganda. One non-inoculated batch was incubated as a reference. Levels of initial and final moisture and cyanogens were assayed. The experiment was done in quadruplicate. Incubation of disinfected root pieces reduced cyanogenic glucoside levels significantly to 62.7% (SD 2.8) of the initial value. Microbial growth resulted in significant additional reduction of the cyanogenic glucoside levels to 29.8% (SD 18.9) of the levels which were obtained after non-inoculated incubation. Among the tested strains, N. sitophila reduced cyanogenic glucoside levels most effectively, followed by R. stolonifer and R. oryzae. Of all fermented samples, both Rhizopus spp. showed highest proportion of residual cyanogens in the cyanohydrin form. Flours showed similar patterns of cyanogens as the batches they were prepared from. Cyanogenic glucoside level reduction was significantly correlated (r = 0.86) with the extent of root softening. It is concluded that both incubation and microbial activity are instrumental in reducing the potential toxicity of cassava during the solid substrate fermentation and that effectiveness varies considerably between the species of microorganisms applied.

Coprophilous fungi of the horse.

PubMed

Pointelli, E; Santa-maria, M A; Caretta, G

1981-05-08

A total of 1267 microfungi, including 35 Myxomycetes, were recorded from the fecal samples of the 60 horses; of these 395 were found on 20 saddle-horse feces, 363 on 20 race-horses and 509 on 20 working horses. Eighty two species representing 53 genera were recorded; of these 7 were Zygomycetes, 18 Ascomycetes, 1 Basidiomycetes and 25 Fungi Imperfecti: 2 Myxomycetes. Common coprophilous fungi are in decreasing order Pilobolus kleinii, Saccobolus depauperatus, Mucor hiemalis, Lasiobolus ciliatus, Podospora curvula, Petriella guttulata, M. circinelloides, Coprinus radiatus, Dictyostelium mucoroides, Sordaria fimicola, C. miser, C. stercorariusm, Acremonium sp., Coprotus granuliformis, Graphium putredinis, Iodophanus carneus, Chaetomium murorum, Podospora communis, P. inaequalis, P. setosa, Saccobolus versicolor and Cladosporium cucumerinum. Species of Myrothecium verrucaria, Actinomucor elegans, Kernia nitida, Spiculostilbella dendritica and Mucor parvispora were found exclusively in working-horses feces. Badhamia sp., Anixiopsis stercoraria, Echinobotryum state of D. stemonitis, Geotrichum candidum and Oidiodendron sp. were found only in saddle-horses feces. Chlamidomyces palmarum, Philocopra sp. were found exclusively in race-horses feces. Notes on infrequent or interesting fungi include Thamnostylum piriforme, Phialocephala dimorphospora, Rhopalomyces elegans and Spiculostilbella dendritica.

[Forefront of diagnosis and treatment of deep-steam mycology in Korea--rhinoorbitocerebral zygomycosis].

PubMed

Lee, Eun Jung; Chung, Jin-Won; Choi, Sangho; Kim, Yang Soo; Woo, Jun Hee

2009-01-01

Mucor is a mold which exists in nature, but mucor infections of humans, even in immunocompromised hosts, are rare. Clinical manifestations of mucormycosis are nonspecific and diagnosis is based on microscopic examination and culture of biopsy specimens. Serologic test or molecular methods of speciation are used only as research tools. We investigated medical records especially for underlying diseases, clinical findings, treatment, and prognosis of patients diagnosed with rhinocerebral mucormycosis retrospectively in the Asan Medical Center. The underlying diseases were diabetes mellitus in 8 patients, acute leukemia in 2, kidney transplantation in 2, and myelodysplastic syndrome in 1 of the total 13 patients. Six patients complained of nasal symptoms including stuffy nose, rhinorrhea, 5 patients complained of ophthalmic symptoms such as decreased visual acuity, diplopia, and ophthalmic pain and 2 of hard palate ulcer. The mortality was 23%(3/13; the two patients with kidney transplant, and one patient with acute leukemia). In summary, mucormycosis should be considered in an uncontrolled DM and an immunocompromised host. The combined modality of early surgical debridement and antifungal agents was used for better treatment of rhinocerebral mucormycosis.

Disseminated mycosis in a killer whale (Orcinus orca).

PubMed

Abdo, Walied; Kawachi, Takeshi; Sakai, Hiroki; Fukushi, Hideto; Kano, Rui; Shibahara, Tomoyuki; Shirouzu, Hiroshi; Kakizoe, Yuko; Tuji, Hajime; Yanai, Tokuma

2012-01-01

Hematological findings in a female killer whale (Orcinus orca) undergoing rehabilitation after sudden severe anorexia revealed continuing increases in serum lactate dehydrogenase and aspartate aminotransferase activities as well as fibrinogen concentration. Serologic evidence of herpesvirus infection and skin vesicles were detected 2 weeks into the treatment regimen of antibiotics and corticosteroids. The whale showed signs of improvement after treatment with anti-herpesvirus drugs, but sudden severe anorexia reappeared, along with marked elevation of fibrinogen concentration that continued until the death. Postmortem examination revealed multiple light tan foci of necrosis in the skeletal and cardiac muscles, and lung consolidation. Microscopic findings indicated disseminated fungal granulomas in the skeletal and cardiac muscles, as well as myocarditis, mycotic embolic thromboarteritis of cardiac blood vessels, and bronchopneumonia with numerous typical Aspergillus-like fungi. Mucor-like structures in granulomas in the heart and skeletal muscle and Aspergillus-like fungi in the lungs were identified using periodic acid-Schiff, Gomori methenamine silver stain, and immunohistochemistry. The present case involves dual infection with Mucor and Aspergillus species in a killer whale with concurrent herpesvirus.

Comparison in antioxidant and antitumor activities of pine polyphenols and its seven biotransformation extracts by fungi

PubMed Central

Li, Hui

2017-01-01

Microbial transformation can strengthen the antioxidant and antitumor activities of polyphenols. Polyphenols contents, antioxidant and antitumor activities of pine polyphenols and its biotransformation extracts by Aspergillus niger, Aspergillus oryzae, Aspergillus carbonarius, Aspergillus candidus, Trichodermas viride, Mucor wutungkiao and Rhizopus sp were studied. Significant differences were noted in antioxidant and antitumor activities. The highest antioxidant activities in Trolox equivalent antioxidant capacity (TEAC), DPPH radical scavenging activity, superoxide anion radical scavenging activity, hydroxyl radical scavenging activity, reducing power assay and antitumor activity against LoVo cells were biotransformation extract of Aspergillus carbonarius (BAC), biotransformation extract of Mucor wutungkiao (BMW), biotransformation extract of Aspergillus carbonarius (BAC), biotransformation extract of Aspergillus niger (BAN), biotransformation extract of Aspergillus oryzae (BAO) and BMW, respectively. Correlation analysis found that antioxidant and antitumor activities were associated with polyphenols contents and types of free radicals and tumors. A. carbonarius can make polyphenol oxidation, hydroxylation and methylation, and form new polyphenols. In conclusion, A. carbonarius, A. niger and M. wutungkiao are valuable microorganisms used for polyphenols biotransformation and enhance the antioxidant and antitumor activities of polyphenols. PMID:28560092

Model of the synthesis of trisporic acid in Mucorales showing bistability.

PubMed

Werner, S; Schroeter, A; Schimek, C; Vlaic, S; Wöstemeyer, J; Schuster, S

2012-12-01

An important substance in the signalling between individuals of Mucor-like fungi is trisporic acid (TA). This compound, together with some of its precursors, serves as a pheromone in mating between (+)- and (-)-mating types. Moreover, intermediates of the TA pathway are exchanged between the two mating partners. Based on differential equations, mathematical models of the synthesis pathways of TA in the two mating types of an idealised Mucor-fungus are here presented. These models include the positive feedback of TA on its own synthesis. The authors compare three sub-models in view of bistability, robustness and the reversibility of transitions. The proposed modelling study showed that, in a system where intermediates are exchanged, a reversible transition between the two stable steady states occurs, whereas an exchange of the end product leads to an irreversible transition. The reversible transition is physiologically favoured, because the high-production state of TA must come to an end eventually. Moreover, the exchange of intermediates and TA is compared with the 3-way handshake widely used by computers linked in a network.

Evaluation of antifungal activity in essential oil of the Syzygium aromaticum (L.) by extraction, purification and analysis of its main component eugenol.

PubMed

Rana, Inder Singh; Rana, Aarti Singh; Rajak, Ram Charan

2011-10-01

Antifungal properties of some essential oils have been well documented. Clove oil is reported to have strong antifungal activity against many fungal species. In this study we have evaluated antifungal potential of essential oil of Syzygium aromaticum (L.) against some common fungal pathogens of plants and animals namely, Fusarium moniliforme NCIM 1100, Fusarium oxysporum MTCC 284, Aspergillus sp., Mucor sp., Trichophyton rubrum and Microsporum gypseum. All fungal species were found to be inhibited by the oil when tested through agar well diffusion method. Minimum inhibitory concentration (MIC) was determined for all the species. Column chromatography was performed to separate the eugenol rich fraction from clove oil. Out of seven fractions maximum activity was obtained in column fraction II. TLC and HPLC data confirmed presence of considerable Eugenol in fraction II and clove oil. Microscopic study on effect of clove oil and column fraction II on spores of Mucor sp. and M. gypseum showed distortion and shrinkage while it was absent in other column fractions. So it can be concluded that the antifungal action of clove oil is due to its high eugenol content.

Biotransformation of Stypotriol triacetate by Aspergillus niger

NASA Astrophysics Data System (ADS)

Areche, Carlos; Vaca, Inmaculada; Labbe, Pamela; Soto-Delgado, Jorge; Astudillo, Luis; Silva, Mario; Rovirosa, Juana; San-Martin, Aurelio

2011-07-01

Biological transformation of the meroditerpenoid, stypotriol triacetate ( 1) by the fungi Aspergillus niger, Cunninghamella elegans, Gibberella fujikuroi and Mucor plumbeus was studied. The incubation of 1 with A. niger yielded the new compound 6',14-diacetoxy-stypol-4,5-dione ( 2) whose structure was established by 1H, 13C and 2D NMR and supported by DFT/GIAO.

Ovicidal, larvicidal and adulticidal properties of Asparagus racemosus (Willd.) (Family: Asparagaceae) root extracts against filariasis (Culex quinquefasciatus), dengue (Aedes aegypti) and malaria (Anopheles stephensi) vector mosquitoes (Diptera: Culicidae).

PubMed

Govindarajan, Marimuthu; Sivakumar, Rajamohan

2014-04-01

Several diseases are associated to the mosquito-human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikungunya fever, West Nile virus and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The present investigation was undertaken to study the ovicidal, larvicidal and adulticidal activities of crude hexane, ethyl acetate, benzene, chloroform and methanol extracts of root of Asparagus racemosus were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi (Diptera: Culicidae). The mean percent hatchability of the eggs was observed after 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the methanol extract showed the highest ovicidal activity. The methanol extract of Asparagus racemosus against Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi exerted 100% mortality (zero hatchability) at 375, 300 and 225 ppm, respectively. Control eggs showed 99-100% hatchability. The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in methanol extract of root of Asparagus racemosus against the larvae of Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi with the LC50 and LC90 values were 115.13, 97.71 and 90.97 ppm and 210.96, 179.92, and 168.82 ppm, respectively. The adult mortality was observed after 24 h recovery period. The plant crude extracts showed dose-dependent mortality. At higher concentrations, the adult showed restless movement for some times with abnormal wagging and then died. Among the extracts tested, the highest adulticidal activity was observed in

Solitary ecthyma gangrenosum (EG)-like lesion consequent to Candida albicans in a neonate.

PubMed

Agarwal, Shilpi; Sharma, Meenal; Mehndirata, Vibhu

2007-06-01

Ecthyma gangrenosum (EG) is a well recognized cutaneous lesion most commonly associated with Pseudomonas aeruginosa bacteremia. It usually occurs in immunocompromized and critically ill patients. Uncommonly, EG like lesions have also been observed in patients with fungal infections, like Fusarium Candida, Mucor and Aspergillus. We present a case of EG- Like lesion due to Candida albicans, in a 12 day old neonate.

Activity of deferasirox in Mucorales: influences of species and exogenous iron.

PubMed

Lewis, Russell E; Pongas, Georgios N; Albert, Nathaniel; Ben-Ami, Ronen; Walsh, Thomas J; Kontoyiannis, Dimitrios P

2011-01-01

Differences in deferasirox susceptibility among members of the order Mucorales are unknown. Here we show that Cunninghamella bertholletiae (four isolates) and Mucor species (five isolates) display higher deferasirox MICs and minimal fungicidal concentrations compared to Rhizopus species (six isolates). Exogenous iron further attenuated the deferasirox susceptibility of Mucorales isolates with low MICs. Vital staining revealed damage to subapical compartments in susceptible strains.

Antagonism of rice phylloplane fungi against Cercospora oryzae

NASA Astrophysics Data System (ADS)

Mardani, A.; Hadiwiyono

2018-03-01

Narrow brown leaf spot (NBLS) caused by Cercospora oryzae Miyake is one of the important obstacle in rice cultivation that can decrease the productivity up to 40%. It has been known well that some phylloplane fungi are antagonistic to some leaf diseases. Phylloplane fungi of rice however haven’t been studied much and poorly understood as biological control agent of rice pathogen such C. oryzae. The research aimed to study the antagonism of some phylloplane fungi of rice against C. oryzae. At least 14 isolates of phylloplane fungi were collected which consisted of six pathogenic and eight nonpathogenic variants. All of nonpathogenic isolates were antagonistic against C. oryzae both in vitro and only one isolate could not inhibit the infection of the pathogen in vivo. Some isolates were identified as Aspergillus, Mucor, Penicillium, Fusarium, and Trichoderma. The isolate of Mucor and Fusarium could inhibit the highest growth of pathogen on potato dextrose medium that were at 36.0% and 35.5% respectively. Whereas on artificial inoculation on rice, some isolates such Penicillium and Fusarium could inhibit most effectively and were significantly different to Mencozeb application with dosage 5g L-1.

Evaluation of antifungal activity in essential oil of the Syzygium aromaticum (L.) by extraction, purification and analysis of its main component eugenol

PubMed Central

Rana, Inder Singh; Rana, Aarti Singh; Rajak, Ram Charan

2011-01-01

Antifungal properties of some essential oils have been well documented. Clove oil is reported to have strong antifungal activity against many fungal species. In this study we have evaluated antifungal potential of essential oil of Syzygium aromaticum (L.) against some common fungal pathogens of plants and animals namely, Fusarium moniliforme NCIM 1100, Fusarium oxysporum MTCC 284, Aspergillus sp., Mucor sp., Trichophyton rubrum and Microsporum gypseum. All fungal species were found to be inhibited by the oil when tested through agar well diffusion method. Minimum inhibitory concentration (MIC) was determined for all the species. Column chromatography was performed to separate the eugenol rich fraction from clove oil. Out of seven fractions maximum activity was obtained in column fraction II. TLC and HPLC data confirmed presence of considerable Eugenol in fraction II and clove oil. Microscopic study on effect of clove oil and column fraction II on spores of Mucor sp. and M. gypseum showed distortion and shrinkage while it was absent in other column fractions. So it can be concluded that the antifungal action of clove oil is due to its high eugenol content. PMID:24031751

Activity of Deferasirox in Mucorales: Influences of Species and Exogenous Iron▿

PubMed Central

Lewis, Russell E.; Pongas, Georgios N.; Albert, Nathaniel; Ben-Ami, Ronen; Walsh, Thomas J.; Kontoyiannis, Dimitrios P.

2011-01-01

Differences in deferasirox susceptibility among members of the order Mucorales are unknown. Here we show that Cunninghamella bertholletiae (four isolates) and Mucor species (five isolates) display higher deferasirox MICs and minimal fungicidal concentrations compared to Rhizopus species (six isolates). Exogenous iron further attenuated the deferasirox susceptibility of Mucorales isolates with low MICs. Vital staining revealed damage to subapical compartments in susceptible strains. PMID:20956598

Enhancement of the functionality of bread by incorporation of Shatavari (Asparagus racemosus).

PubMed

Singh, Nishu; Jha, Alok; Chaudhary, Anand; Upadhyay, Ashutosh

2014-09-01

In view of the wider consumption of bakery products, they could be good choice for the delivery of functionality. The present study attempts to develop a functional formulation of bread by incorporation of shatavari (Asparagus racemosus Willd.), which is an important medicinal plant of India. Central composite rotatable design (CCRD) was used for experiments in which yeast and shatavari powder were taken as variables. Response surface methodology (RSM) was used to optimize the bread formulations on the basis of hardness, adhesiveness, springiness, chewiness and cohesiveness as responses. Qualitative tests were performed for assessing the presence of phytochemicals in shatavari bread. Sensory attributes of the shatavari bread were evaluated using descriptive analysis technique. The optimum acceptable level for shatavari and yeast in bread was found to be 3.5 % and 4.96 %, respectively. All the phytochemicals such as alkaloid, steroid, terpenoid and saponin present in original herbs were also present in bread. However flavonoids were not found in the bread when analysed qualitatively and using TLC.

Advantages of Environmental Scanning Electron Microscopy in Studies of Microorganisms

DTIC Science & Technology

1993-01-01

external structures, cultures of Aspergillus sp. and Cunninghamella sp. but details of cytoplasmic organization were lacking were maintained on corn meal ... agar . (Fig. 7). Uncoated, freeze-dried Euglena sp. cells exam- ined with either ESD or BSE detectors did not exhibit Specimen Preparation for ESEM...approximately Agar plugs from sporulating cultures of Cunning- 1.0 pLm in diameter (Fig. 23). Many of the vesicles ap- hamella sp., Mucor sp., and Aspergillus

Activity of posaconazole and other antifungal agents against Mucorales strains identified by sequencing of internal transcribed spacers.

PubMed

Alastruey-Izquierdo, Ana; Castelli, Maria Victoria; Cuesta, Isabel; Monzon, Araceli; Cuenca-Estrella, Manuel; Rodriguez-Tudela, Juan Luis

2009-04-01

The antifungal susceptibility profiles of 77 clinical strains of Mucorales species, identified by internal transcribed spacer sequencing, were analyzed. MICs obtained at 24 and 48 h were compared. Amphotericin B was the most active agent against all isolates, except for Cunninghamella and Apophysomyces isolates. Posaconazole also showed good activity for all species but Cunninghamella bertholletiae. Voriconazole had no activity against any of the fungi tested. Terbinafine showed good activity, except for Rhizopus oryzae, Mucor circinelloides, and Rhizomucor variabilis isolates.

Influence of growth regulators and elicitors on cell growth and α-tocopherol and pigment productions in cell cultures of Carthamus tinctorius L.

PubMed

Chavan, Smita P; Lokhande, Vinayak H; Nitnaware, Kirti M; Nikam, Tukaram D

2011-03-01

The present study examined the effects of plant growth hormones, incubation period, biotic (Trametes versicolor, Mucor sp., Penicillium notatum, Rhizopus stolonifer, and Fusarium oxysporum) and abiotic (NaCl, MgSO(4), FeSO(4), ZnSO(4), and FeCl(3)) elicitors on cell growth and α-tocopherol and pigment (red and yellow) productions in Carthamus tinctorius cell cultures. The cell growth and α-tocopherol and pigment contents improved significantly on Murashige and Skoog (MS) liquid medium containing 50.0 μM α-naphthalene acetic acid (NAA) and 2.5 μM 6-Benzyladenine (BA) at 28 days of incubation period. Incorporation of T. versicolor (50 mg l(-1)) significantly enhanced the production of α-tocopherol (12.7-fold) and red pigment (4.24-fold). Similarly, supplementation of 30 mg l(-1) T. versicolor (7.54-fold) and 70 mg l(-1) Mucor sp. (7.40-fold) significantly increased the production of yellow pigment. Among abiotic elicitors, NaCl (50-70 mg l(-1)) and MgSO(4) (10-30 mg l(-1)) significantly improved production of α-tocopherol (1.24-fold) and red pigment (20-fold), whereas yellow pigment content increased considerably by all the abiotic elicitor treatments. Taken together, the present study reports improved productions of α-tocopherol and the pigment as a stress response of safflower cell cultures exposed to these elicitors.

Identification of Mucorales From Clinical Specimens: A 4-Year Experience in a Single Institution

PubMed Central

Yang, Mina; Lee, Jang Ho; Kim, Young-Kwon; Ki, Chang-Seok

2016-01-01

Mucormycosis, a fatal opportunistic infection in immunocompromised hosts, is caused by fungi belonging to the order Mucorales. Early diagnosis based on exact identification and multidisciplinary treatments is critical. However, identification of Mucorales fungi is difficult and often delayed, resulting in poor prognosis. This study aimed to compare the results of phenotypic and molecular identification of 12 Mucorales isolates collected from 4-yr-accumulated data. All isolates were identified on the basis of phenotypic characteristics such as growth rate, colony morphology, and reproductive structures. PCR and direct sequencing were performed to target internal transcribed spacer (ITS) and/or D1/D2 regions. Target DNA sequencing identified five Lichtheimia isolates, two Rhizopus microsporus isolates, two Rhizomucor pusillus isolates, one Cunninghamella bertholletiae isolate, one Mucor fragilis isolate, and one Syncephalastrum racemosum isolate. Five of the 12 (41.7%) isolates were incorrectly identified on the basis of phenotypic identification. DNA sequencing showed that of these five isolates, two were Lichtheimia isolates, one was Mucor isolate, one was Rhizomucor isolate, and one was Rhizopus microspores. All the isolates were identified at the species level by ITS and/or D1/D2 analyses. Phenotypic differentiation and identification of Mucorales is difficult because different Mucorales share similar morphology. Our results indicate that the molecular methods employed in this study are valuable for identifying Mucorales. PMID:26522761

Identification of mucorales from clinical specimens: a 4-year experience in a single institution.

PubMed

Yang, Mina; Lee, Jang Ho; Kim, Young Kwon; Ki, Chang Seok; Huh, Hee Jae; Lee, Nam Yong

2016-01-01

Mucormycosis, a fatal opportunistic infection in immunocompromised hosts, is caused by fungi belonging to the order Mucorales. Early diagnosis based on exact identification and multidisciplinary treatments is critical. However, identification of Mucorales fungi is difficult and often delayed, resulting in poor prognosis. This study aimed to compare the results of phenotypic and molecular identification of 12 Mucorales isolates collected from 4-yr-accumulated data. All isolates were identified on the basis of phenotypic characteristics such as growth rate, colony morphology, and reproductive structures. PCR and direct sequencing were performed to target internal transcribed spacer (ITS) and/or D1/D2 regions. Target DNA sequencing identified five Lichtheimia isolates, two Rhizopus microsporus isolates, two Rhizomucor pusillus isolates, one Cunninghamella bertholletiae isolate, one Mucor fragilis isolate, and one Syncephalastrum racemosum isolate. Five of the 12 (41.7%) isolates were incorrectly identified on the basis of phenotypic identification. DNA sequencing showed that of these five isolates, two were Lichtheimia isolates, one was Mucor isolate, one was Rhizomucor isolate, and one was Rhizopus microspores. All the isolates were identified at the species level by ITS and/or D1/D2 analyses. Phenotypic differentiation and identification of Mucorales is difficult because different Mucorales share similar morphology. Our results indicate that the molecular methods employed in this study are valuable for identifying Mucorales.

Fungal infection by Mucorales order in lung transplantation: 4 case reports.

PubMed

Neto, F M F D; Camargo, P C L B; Costa, A N; Teixeira, R H O B; Carraro, R M; Afonso, J E; Campos, S V; Samano, M N; Fernandes, L M; Abdalla, L G; Pêgo-Fernandes, P M

2014-01-01

Mucorales is a fungus that causes systemic, highly lethal infections in immunocompromised patients. The overall mortality of pulmonary mucormycosis can reach 95%. This work is a review of medical records of 200 lung transplant recipients between the years of 2003 and 2013, in order to identify the prevalence of Mucorales in the Lung Transplantation service of Heart Institute (InCor), Hospital das Clínicas da Universidade de São Paulo, Brazil, by culture results from bronchoalveolar lavage and necropsy findings. We report 4 cases found at this analyses: 3 in patients with cystic fibrosis and 1 in a patient with bronchiectasis due to Kartagener syndrome. There were 2 unfavorable outcomes related to the presence of Mucorales, 1 by reduction of immunosuppression, another by invasive infection. Another patient died from renal and septic complications from another etiology. One patient was diagnosed at autopsy just 5 days after lung transplantation, with the Mucor inside the pulmonary vein with a precise, well-defined involvement only of donor's segment, leading to previous colonization hypothesis. There are few case reports of Mucorales infection in lung transplantation in the literature. Surveillance for the presence of Mucor can lead to timely fungal treatment and reduce morbidity and mortality in the immunocompromised patients, especially lung transplant recipients. Copyright © 2014 Elsevier Inc. All rights reserved.

Biotechnological Applications of Dimorphic Yeasts

NASA Astrophysics Data System (ADS)

Doiphode, N.; Joshi, C.; Ghormade, V.; Deshpande, M. V.

The dimorphic yeasts have the equilibrium between spherical growth (budding) and polarized (hyphal or pseudohyphal tip elongation) which can be triggered by change in the environmental conditions. The reversible growth phenomenon has made dimorphic yeasts as an useful model to understand fungal evolution and fungal differentiation, in general. In nature dimorphism is clearly evident in plant and animal fungal pathogens, which survive and most importantly proliferate in the respective hosts. However, number of organisms with no known pathogenic behaviour also show such a transition, which can be exploited for the technological applications due to their different biochemical make up under different morphologies. For instance, chitin and chitosan production using dimorphic Saccharomyces, Mucor, Rhizopus and Benjaminiella, oil degradation and biotransformation with yeast-form of Yarrowia species, bioremediation of organic pollutants, exopolysac-charide production by yeast-phase of Aureobasidium pullulans, to name a few. Myrothecium verrucaria can be used for seed dressing in its yeast form and it produces a mycolytic enzyme complex in its hyphal-form for the biocontrol of fungal pathogens, while Beauveria bassiana and other entomopathogens kill the insect pest by producing yeast- like cells in the insect body. The form-specific expression of protease, chitinase, lipase, ornithine decarboxylase, glutamate dehydrogenases, etc. make Benjaminiella poitrasii, Basidiobolus sp., and Mucor rouxii strains important in bioremediation, nanobiotechnology, fungal evolution and other areas.

Bioconversion of straw into improved fodder: fungal flora decomposing rice straw.

PubMed

Helal, G A

2005-09-01

The fungal flora decomposing rice straw were investigated all over the soil of Sharkia Province, east of Nile Delta, Egypt, using the nylon net bag technique. Sixty-four straw-decomposing species belonging to 30 genera were isolated by the dilution plate method in ground rice straw-Czapek's agar medium at pH 6. The plates were incubated separately at 5℃, 25℃ and 45℃, respectively. Twenty nine species belonging to 14 genera were isolated at 5℃. The most frequent genus was Penicillium (seven species), and the next frequent genera were Acremonium (three species), Fusarium (three species), Alternaria, Chaetomium, Cladosporium, Mucor, Stachybotrys (two species) and Rhizopus stolonifer. At 25℃, 47 species belonging to 24 genera were isolated. The most frequent genus was Aspergillus (nine species), and the next frequent genera were ranked by Penicillium (five species), Chaetomium (three species), Fusarium (three species). Each of Alternaria, Cladosporium, Mucor, Myrothecium and Trichoderma was represented by two species. At 45℃, 15 species belonging to seven genera were isolated. These were seven species of Aspergillus, two species of Chaetomium and two species of Emericella, while Humicola, Malbranchea, Rhizomucor and Talaromyces were represented by one species respectively. The total counts of fungi the genera, and species per gram of dry straw were significantly affected by incubation temperature and soil analysis (P < 0.05).

Activity of Posaconazole and Other Antifungal Agents against Mucorales Strains Identified by Sequencing of Internal Transcribed Spacers▿

PubMed Central

Alastruey-Izquierdo, Ana; Castelli, Maria Victoria; Cuesta, Isabel; Monzon, Araceli; Cuenca-Estrella, Manuel; Rodriguez-Tudela, Juan Luis

2009-01-01

The antifungal susceptibility profiles of 77 clinical strains of Mucorales species, identified by internal transcribed spacer sequencing, were analyzed. MICs obtained at 24 and 48 h were compared. Amphotericin B was the most active agent against all isolates, except for Cunninghamella and Apophysomyces isolates. Posaconazole also showed good activity for all species but Cunninghamella bertholletiae. Voriconazole had no activity against any of the fungi tested. Terbinafine showed good activity, except for Rhizopus oryzae, Mucor circinelloides, and Rhizomucor variabilis isolates. PMID:19171801

Chemo-enzymatic synthesis of a glycosylated peptide containing a complex N-glycan based on unprotected oligosaccharides by using DMT-MM and Endo-M.

PubMed

Tomabechi, Yusuke; Katoh, Toshihiko; Kunishima, Munetaka; Inazu, Toshiyuki; Yamamoto, Kenji

2017-08-01

For chemo-enzymatic synthesis of a glycosylated peptide, 4-(4,6-dimethoxy-1,3,5-triazin-2-yl)-4-methylmorpholinium chloride (DMT-MM) was used for the synthesis of a N-acetylglucosaminyl peptide and a pseudoglycopeptide by solid-phase peptide synthesis without the requirement of protecting groups on the carbohydrate. We also performed transglycosylation of an N-glycan to the N-acetylglucosaminyl peptide using endo-β-N-acetylglucosaminidase from Mucor hiemalis (Endo-M) to synthesize a glycopeptide containing a complex N-glycan.

Uncommon opportunistic fungal infections of oral cavity: A review

PubMed Central

Deepa, AG; Nair, Bindu J; Sivakumar, TT; Joseph, Anna P

2014-01-01

The majority of opportunistic oral mucosal fungal infections are due to Candida albicans and Aspergillus fumigatus species. Mucor and Cryptococcus also have a major role in causing oral infections, whereas Geotrichum, Fusarium, Rhodotorula, Saccharomyces and Penicillium marneffei are uncommon pathogens in the oral cavity. The broad spectrum of clinical presentation includes pseudo-membranes, abscesses, ulcers, pustules and extensive tissue necrosis involving bone. This review discusses various uncommon opportunistic fungal infections affecting the oral cavity including their morphology, clinical features and diagnostic methods. PMID:25328305

Detoxification of fusaric acid by the soil microbe Mucor rouxii

USDA-ARS?s Scientific Manuscript database

An unusually aggressive biotype of the root rotting pathogen of cotton, Fusarium oxysporum f. sp. vasinfectum (Fov), has been identified in the Western Hemisphere in some cotton fields in California. This pathogen produces copious quantities of the plant toxin fusaric acid (5-butyl-2-pyridinecarbox...

Phytate degradation by fungi and bacteria that inhabit sawdust and coffee residue composts.

PubMed

Fathallh Eida, Mohamed; Nagaoka, Toshinori; Wasaki, Jun; Kouno, Kenji

2013-01-01

Phytate is the primary source of organic phosphorus, but it cannot be directly utilized by plants and is strongly adsorbed by the soil, reducing bioavailability. Composting is a process used to improve the bioavailability of phytate in organic wastes through degradation by microorganisms. In this study, we aimed to investigate the phytate-degrading ability of fungi and bacteria that inhabit sawdust compost and coffee residue compost, and their contribution to the composting process. In the plate assay, the fungi that formed clear zones around their colonies belonged to the genera Mucor, Penicillium, Galactomyces, Coniochaeta, Aspergillus, and Fusarium, while the bacteria belonged to the genera Pseudomonas, Enterobacter, Chitinophaga, and Rahnella. Eight fungal isolates (genera Mucor, Penicillium, Galactomyces, and Coniochaeta) and four bacterial isolates (genera Pseudomonas, Enterobacter, and Rahnella) were selected to evaluate phytase activity in their liquid culture and their ability to degrade phytate in organic materials composed of mushroom media residue and rice bran. The selected fungi degraded phytate in organic materials to varying degrees. Penicillium isolates showed the highest degradation ability and Coniochaeta isolate exhibited relatively high degradation ability. The clear zone diameters of these fungal isolates displayed significantly positive and negative correlations with inorganic and phytate phosphorus contents in the organic materials after incubation, respectively; however, none of the selected bacteria reduced phytate phosphorus in organic materials. It is therefore possible that fungi are major contributors to phytate degradation during composting.

Phytate Degradation by Fungi and Bacteria that Inhabit Sawdust and Coffee Residue Composts

PubMed Central

Eida, Mohamed Fathallh; Nagaoka, Toshinori; Wasaki, Jun; Kouno, Kenji

2013-01-01

Phytate is the primary source of organic phosphorus, but it cannot be directly utilized by plants and is strongly adsorbed by the soil, reducing bioavailability. Composting is a process used to improve the bioavailability of phytate in organic wastes through degradation by microorganisms. In this study, we aimed to investigate the phytate-degrading ability of fungi and bacteria that inhabit sawdust compost and coffee residue compost, and their contribution to the composting process. In the plate assay, the fungi that formed clear zones around their colonies belonged to the genera Mucor, Penicillium, Galactomyces, Coniochaeta, Aspergillus, and Fusarium, while the bacteria belonged to the genera Pseudomonas, Enterobacter, Chitinophaga, and Rahnella. Eight fungal isolates (genera Mucor, Penicillium, Galactomyces, and Coniochaeta) and four bacterial isolates (genera Pseudomonas, Enterobacter, and Rahnella) were selected to evaluate phytase activity in their liquid culture and their ability to degrade phytate in organic materials composed of mushroom media residue and rice bran. The selected fungi degraded phytate in organic materials to varying degrees. Penicillium isolates showed the highest degradation ability and Coniochaeta isolate exhibited relatively high degradation ability. The clear zone diameters of these fungal isolates displayed significantly positive and negative correlations with inorganic and phytate phosphorus contents in the organic materials after incubation, respectively; however, none of the selected bacteria reduced phytate phosphorus in organic materials. It is therefore possible that fungi are major contributors to phytate degradation during composting. PMID:23100024

Utility of intraoperative frozen sections in surgical decision making for acute invasive fungal rhinosinusitis.

PubMed

Papagiannopoulos, Peter; Lin, Diana Murro; Al-Khudari, Samer; Rajan, Kumar; Reddy, Swathi; Gattuso, Paulo; Tajudeen, Bobby; Batra, Pete S

2017-05-01

Acute invasive fungal rhinosinusitis (AIFRS) represents a fulminant, potentially fatal, disease process in immunocompromised patients. The diagnosis often rests on high index of clinical suspicion, with relative paucity of data on the diagnostic and therapeutic implications of intraoperative frozen sections. Retrospective review was performed for 18 cases undergoing endoscopic sinus surgery for AIFRS. Reliability of intraoperative frozen section diagnosis was evaluated for all patients using final pathology as the gold standard. A total of 66 frozen sections were performed. Diagnostic accuracy of frozen sections illustrated sensitivity of 72.7% (95% confidence interval [CI], 0.57 to 0.85), specificity of 100% (95% CI, 0.85 to 1.00), positive predictive value (PPV) of 100% (95% CI, 0.89 to 1.00), and negative predictive value (NPV) of 64.7% (95% CI, 0.46 to 0.80). There was no statistically significant difference in sensitivity of frozen sections in cases of Mucor and Aspergillus at 68.8%% and 76.2%, respectively (p = 0.61). This study represents the largest series assessing the diagnostic accuracy of frozen section analysis in AIFRS. Frozen section analysis is an effective tool for guiding intraoperative decision making in patients with AIFRS with a high PPV. A Low NPV underscores the importance of clinical suspicion and intraoperative decision making based on endoscopic findings when negative frozen section results are encountered. Further, frozen section analysis appears to be equally effective in detecting either Mucor or Aspergillus. © 2017 ARS-AAOA, LLC.

Fungal outbreak in a show cave.

PubMed

Jurado, V; Porca, E; Cuezva, S; Fernandez-Cortes, A; Sanchez-Moral, S; Saiz-Jimenez, C

2010-08-01

Castañar de Ibor Cave (Spain) was discovered in 1967 and declared a Natural Monument in 1997. In 2003 the cave was opened to public visits. Despite of extensive control, on 26 August 2008 the cave walls and sediments appeared colonized by long, white fungal mycelia. This event was the result of an accidental input of detritus on the afternoon of 24 August 2008. We report here a fungal outbreak initiated by Mucor circinelloides and Fusarium solani and the methods used to control it. 2010 Elsevier B.V. All rights reserved.

Impact of stress on female reproductive health disorders: Possible beneficial effects of shatavari (Asparagus racemosus).

PubMed

Pandey, Ajai K; Gupta, Anumegha; Tiwari, Meenakshi; Prasad, Shilpa; Pandey, Ashutosh N; Yadav, Pramod K; Sharma, Alka; Sahu, Kankshi; Asrafuzzaman, Syed; Vengayil, Doyil T; Shrivastav, Tulsidas G; Chaube, Shail K

2018-07-01

Stress is deeply rooted in the society and women are frequently exposed to psychological, physical and physiological stressors. Psychological stress disturbs reproductive health by inducing generation of reactive oxygen species (ROS) and thereby oxidative stress (OS). The increased OS may affect physiology of ovary, oocyte quality and cause female reproductive health disorders. To overcome stress-mediated reproductive health disorders in women, shatavari (Asparagus racemosus) is frequently recommended in Ayurvedic system of medicine. Although shatavari is one of the major health tonics and most popular rasayana drugs to treat reproductive ailments of women, underlying mechanism of shatavari action at the level of ovary remains poorly understood. Based on the existing studies, we propose that shatavari may improve female reproductive health complications including hormonal imbalance, polycystic ovarian syndrome (PCOS), follicular growth and development, oocyte quality and infertility possibly by reducing OS level and increasing antioxidants level in the body. Further studies are required to elucidate the mechanism of shatavari actions at the level of ovary and oocyte that directly impacts the reproductive health of women. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Quantitative polymerase chain reaction detection of circulating DNA in serum for early diagnosis of mucormycosis in immunocompromised patients.

PubMed

Millon, Laurence; Larosa, Fabrice; Lepiller, Quentin; Legrand, Faezeh; Rocchi, Steffi; Daguindau, Etienne; Scherer, Emeline; Bellanger, Anne-Pauline; Leroy, Joel; Grenouillet, Frederic

2013-05-01

The aim of our study was to assess the detection of circulating DNA from the most common species of Mucorales for early diagnosis of mucormycosis in at-risk patients. We retrospectively evaluated a combination of 3 quantitative polymerase chain reaction (qPCR) assays using hydrolysis probes targeting Mucor/Rhizopus, Lichtheimia (formerly Absidia), and Rhizomucor for circulating Mucorales detection. Serial serum samples from 10 patients diagnosed with proven mucormycosis (2-9 samples per patient) were analyzed. No cross-reactivity was detected in the 3 qPCR assays using 19 reference strains of opportunistic fungi, and the limit of detection ranged from 3.7 to 15 femtograms/10 µL, depending on the species. DNA from Mucorales was detected in the serum of 9 of 10 patients between 68 and 3 days before mucormycosis diagnosis was confirmed by histopathological examination and/or positive culture. All the qPCR results were concordant with culture and/or PCR-based identification of the causing agents in tissue (Lichtheimia species, Rhizomucor species, and Mucor/Rhizopus species in 4, 3, and 2 patients, respectively). Quantitative PCR was negative in only 1 patient with proven disseminated mucormycosis caused by Lichtheimia species. Our study suggests that using specific qPCR targeting several species of Mucorales according to local ecology to screen at-risk patients could be useful in a clinical setting. The cost and efficacy of this strategy should be evaluated. However, given the human and economic cost of mucormycosis and the need for rapid diagnosis to initiate prompt directed antifungal therapy, this strategy could be highly attractive.

Search for fungi-specific metabolites of four model drugs in postmortem blood as potential indicators of postmortem fungal metabolism.

PubMed

Martínez-Ramírez, Jorge A; Strien, Juliane; Walther, Grit; Peters, Frank T

2016-05-01

Fungi colonizing cadavers are capable of drug metabolism and may thus change the metabolite pattern or concentration of drugs in forensic postmortem samples. The purpose of this study was to check for the presence of such changes by searching fungi-specific metabolites of four model drugs (amitriptyline, metoprolol, mirtazapine, and zolpidem) in decomposed postmortem blood samples from 33 cases involving these drugs. After isolation and identification of fungal strains present in the samples, each isolate was incubated in Sabouraud medium at 25°C for up to 120h with each model drug. One part of the supernatants was directly analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS), another after liquid-liquid extraction with chlorobutane and concentration. From 21 out of 33 decomposed postmortem blood samples (64%) a total of 30 different strains could be isolated, one from the class of Ascomycete and the rest belonging to 15 species from 8 different genera (number of species): Aspergillus (2), Botrytis (1), Candida (8), Fusarium (1), Mucor (1), Penicillium (1), and Rodothorula (1). In the in vitro studies, these microorganisms were found capable of N-demethylation and N-oxidation of amitriptyline and mirtazapine, O-demethylation followed by side chain oxidation of metoprolol as well as hydroxylation of all four-model drugs. In two of the postmortem blood samples, from which the fungi Aspergillus jensenii, Candida parapsilosis. and Mucor circinelloides had been isolated, a fungi-specific hydroxy zolpidem metabolite was detected. The presence of this metabolite in postmortem samples likely indicates postmortem fungal biodegradation. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Growth of chitinolytic dune soil beta-subclass Proteobacteria in response to invading fungal hyphae.

PubMed

De Boer, W; Klein Gunnewiek, P J; Kowalchuk, G A; Van Veen, J A

2001-08-01

It has frequently been reported that chitinolytic soil bacteria, in particular biocontrol strains, can lyse living fungal hyphae, thereby releasing potential growth substrate. However, the conditions used in such assays (high bacterial density, rich media, fragmented hyphae) make it difficult to determine whether mycolytic activity is actually of importance for the growth and survival of chitinolytic bacteria in soils. An unidentified group of beta-subclass Proteobacteria (CbetaPs) was most dominant among the culturable nonfilamentous chitinolytic bacteria isolated from Dutch sand dune soils. Here we demonstrate that the CbetaPs grew at the expense of extending fungal mycelium of three dune soil fungi (Chaetomium globosum, Fusarium culmorum, and Mucor hiemalis) under nutrient-limiting, soil-like conditions. Aggregates of CbetaPs were also often found attached to fungal hyphae. The growth of a control group of dominant nonchitinolytic dune soil bacteria (beta- and gamma-subclass Proteobacteria) was not stimulated in the mycelial zone, indicating that growth-supporting materials were not independently released in appreciable amounts by the extending hyphae. Therefore, mycolytic activities of CbetaPs have apparently been involved in allowing them to grow after exposure to living hyphae. The chitinase inhibitor allosamidin did not, in the case of Mucor, or only partially, in the cases of Chaetomium and Fusarium, repress mycolytic growth of the CbetaPs, indicating that chitinase activity alone could not explain the extent of bacterial proliferation. Chitinolytic Stenotrophomonas-like and Cytophaga-like bacteria, isolated from the same dune soils, were only slightly stimulated by exposure to fungal hyphae.

Antifungal activity of 1'-homo-N-1,2,3-triazol-bicyclic carbonucleosides: A novel type of compound afforded by azide-enolate (3+2) cycloaddition.

PubMed

González-Calderón, Davir; Mejía-Dionicio, María G; Morales-Reza, Marco A; Aguirre-de Paz, José G; Ramírez-Villalva, Alejandra; Morales-Rodríguez, Macario; Fuentes-Benítes, Aydeé; González-Romero, Carlos

2016-12-01

The first report of 1'-homo-N-1,2,3-triazol-bicyclic carbonucleosides (7a and 7b) is described herein. Azide-enolate (3+2) cycloaddition afforded the synthesis of this novel type of compound. Antifungal activity was evaluated in vitro against four filamentous fungi (Aspergillus fumigatus, Trichosporon cutaneum, Rhizopus oryzae and Mucor hiemalis) as well as nine species of Candida spp. as yeast specimens. These pre-clinical studies suggest that compounds 7a and 7b are promising candidates for complementary biological studies due to their good activity against Candida spp. Copyright © 2016 Elsevier Inc. All rights reserved.

Herbicide Orange Site Treatment and Environmental Monitoring: Summary Report and Recommendations for Naval Construction Battalion Center, Gulfport, Mississippi.

DTIC Science & Technology

1979-11-01

Aspergillus spp. X X Penicillium spp. X X X X Cunninghamella spp. X X Zygorhynchus sp . X X Alternaria sp . X X Mycelial Molds X X Candida spp. X X...itermined if a technique could he used t dtrmn , ’ old s 1 from new sp i 11 . 12 it was not leaving the base and most likely was being held in the bottom...Rhodotorula sp . X X X Geotrichum sp . X X Trichoderma spp. X X X Mucor spp. X X X Rhizopus sp . X X Absidia sp . X X aMicrobial analyses conducted by Department

Discovery of tanshinone derivatives with anti-MRSA activity via targeted bio-transformation.

PubMed

He, Wenni; Liu, Miaomiao; Huang, Pei; Abdel-Mageed, Wael M; Han, Jianying; Watrous, Jeramie D; Nguyen, Don D; Wang, Wenzhao; Song, Fuhang; Dai, Huanqin; Zhang, Jingyu; Quinn, Ronald J; Grkovi, Tanja; Luo, Houwei; Zhang, Lixin; Liu, Xueting

2016-09-01

Two potent anti-MRSA tanshinone glycosides ( 1 and 2 ) were discovered by targeted microbial biotransformation, along with rapid identification via MS/MS networking. Serial reactions including dehydrogenation, demethylations, reduction, glycosylation and methylation have been observed after incubation of tanshinone IIA and fungus Mucor rouxianus AS 3.3447. In addition, tanshinosides B ( 2 ) showed potent activities against serial clinical isolates of oxacillin-resistant Staphylococcus aureus with MIC values of 0.78 μg/mL. This is the first study that shows a significant increase in the level and activities of tanshinone glycosides relative to the substrate tanshinone IIA.

In Vitro Activity of Isavuconazole and Comparators against Clinical Isolates of the Mucorales Order

PubMed Central

Jensen, Rasmus Hare; Meletiadis, Joseph

2015-01-01

The in vitro activity of isavuconazole against Mucorales isolates measured by EUCAST E.Def 9.2 and CLSI M38-A2 methodologies was investigated in comparison with those of amphotericin B, posaconazole, and voriconazole. Seventy-two isolates were included: 12 of Lichtheimia corymbifera, 5 of Lichtheimia ramosa, 5 of group I and 9 of group II of Mucor circinelloides, 9 of Rhizomucor pusillus, 26 of Rhizopus microsporus, and 6 of Rhizopus oryzae. Species identification was confirmed by internal transcribed spacer (ITS) sequencing. EUCAST MICs were read on day 1 (EUCAST-d1) and day 2 (EUCAST-d2), and CLSI MICs were read on day 2 (CLSI-d2). Isavuconazole MIC50s (range) (mg/liter) by EUCAST-d1, CLSI-d2, and EUCAST-d2 were 1 (0.125 to 16), 1 (0.125 to 2), and 4 (0.5 to >16), respectively, across all isolates. The similar values for comparator drugs were as follows: posaconazole, 0.25 (≤0.03 to >16), 0.25 (0.06 to >16), and 1 (0.06 to >16); amphotericin, 0.06 (≤0.03 to 0.5), 0.06 (≤0.03 to 0.25), and 0.125 (≤0.03 to 1); voriconazole, 16 (2 to >16), 8 (1 to >16), and >16 (8 to >16), respectively. Isavuconazole activity varied by species: Lichtheimia corymbifera, 1 (0.5 to 2), 1 (1 to 2), and 2 (1 to 4); Lichtheimia ramosa, 0.25 (0.125 to 0.5), 1 (0.5 to 2), and 2 (0.5 to 4); Rhizomucor pusillus, 0.5 (0.5 to 1), 1 (0.125 to 1), and 2 (1 to 2); Rhizopus microsporus, 1 (0.5 to 4), 0.5 (0.125 to 1), and 4 (1 to 8); and Rhizopus oryzae, 1 (0.5 to 4), 1 (0.125 to 2), and 4 (0.5 to 8), respectively, were more susceptible than Mucor circinelloides: group I, 8 (4 to 8), 4 (2 to 4), and 16 (2 to 16), respectively, and group II, 8 (1 to 16), 8 (1 to 8), and 16 (4 to >16), respectively. This was also observed for posaconazole. The essential agreement was best between EUCAST-d1 and CLSI-d2 (75% to 83%). Isavuconazole displayed in vitro activity against Mucorales isolates with the exception of Mucor circinelloides. The MICs were in general 1 to 3 steps higher than those for

In Vitro Activity of Isavuconazole and Comparators against Clinical Isolates of the Mucorales Order.

PubMed

Arendrup, Maiken Cavling; Jensen, Rasmus Hare; Meletiadis, Joseph

2015-12-01

The in vitro activity of isavuconazole against Mucorales isolates measured by EUCAST E.Def 9.2 and CLSI M38-A2 methodologies was investigated in comparison with those of amphotericin B, posaconazole, and voriconazole. Seventy-two isolates were included: 12 of Lichtheimia corymbifera, 5 of Lichtheimia ramosa, 5 of group I and 9 of group II of Mucor circinelloides, 9 of Rhizomucor pusillus, 26 of Rhizopus microsporus, and 6 of Rhizopus oryzae. Species identification was confirmed by internal transcribed spacer (ITS) sequencing. EUCAST MICs were read on day 1 (EUCAST-d1) and day 2 (EUCAST-d2), and CLSI MICs were read on day 2 (CLSI-d2). Isavuconazole MIC50s (range) (mg/liter) by EUCAST-d1, CLSI-d2, and EUCAST-d2 were 1 (0.125 to 16), 1 (0.125 to 2), and 4 (0.5 to >16), respectively, across all isolates. The similar values for comparator drugs were as follows: posaconazole, 0.25 (≤ 0.03 to >16), 0.25 (0.06 to >16), and 1 (0.06 to >16); amphotericin, 0.06 (≤ 0.03 to 0.5), 0.06 (≤ 0.03 to 0.25), and 0.125 (≤ 0.03 to 1); voriconazole, 16 (2 to >16), 8 (1 to >16), and >16 (8 to >16), respectively. Isavuconazole activity varied by species: Lichtheimia corymbifera, 1 (0.5 to 2), 1 (1 to 2), and 2 (1 to 4); Lichtheimia ramosa, 0.25 (0.125 to 0.5), 1 (0.5 to 2), and 2 (0.5 to 4); Rhizomucor pusillus, 0.5 (0.5 to 1), 1 (0.125 to 1), and 2 (1 to 2); Rhizopus microsporus, 1 (0.5 to 4), 0.5 (0.125 to 1), and 4 (1 to 8); and Rhizopus oryzae, 1 (0.5 to 4), 1 (0.125 to 2), and 4 (0.5 to 8), respectively, were more susceptible than Mucor circinelloides: group I, 8 (4 to 8), 4 (2 to 4), and 16 (2 to 16), respectively, and group II, 8 (1 to 16), 8 (1 to 8), and 16 (4 to >16), respectively. This was also observed for posaconazole. The essential agreement was best between EUCAST-d1 and CLSI-d2 (75% to 83%). Isavuconazole displayed in vitro activity against Mucorales isolates with the exception of Mucor circinelloides. The MICs were in general 1 to 3 steps higher than those for

Protease and lipase activities of fungal and bacterial strains derived from an artisanal raw ewe's milk cheese.

PubMed

Ozturkoglu-Budak, Sebnem; Wiebenga, Ad; Bron, Peter A; de Vries, Ronald P

2016-11-21

We previously identified the microbiota present during cheese ripening and observed high protease and lipase activity in Divle Cave cheese. To determine the contribution of individual isolates to enzyme activities, we investigated a range of species representing this microbiota for their proteolytic and lipolytic ability. In total, 17 fungal, 5 yeast and 18 bacterial strains, previously isolated from Divle Cave cheese, were assessed. Qualitative protease and lipase activities were performed on skim-milk agar and spirit-blue lipase agar, respectively, and resulted in a selection of strains for quantitative assays. For the quantitative assays, the strains were grown on minimal medium containing irradiated Divle Cave cheese, obtained from the first day of ripening. Out of 16 selected filamentous fungi, Penicillium brevicompactum, Penicillium cavernicola and Penicillium olsonii showed the highest protease activity, while Mucor racemosus was the best lipase producer. Yarrowia lipolytica was the best performing yeast with respect to protease and lipase activity. From the 18 bacterial strains, 14 and 11 strains, respectively showed protease and lipase activity in agar plates. Micrococcus luteus, Bacillus stratosphericus, Brevibacterium antiquum, Psychrobacter glacincola and Pseudomonas proteolytica displayed the highest protease and lipase activity. The proteases of yeast and filamentous fungi were identified as mainly aspartic protease by specific inhibition with Pepstatin A, whereas inhibition by PMSF (phenylmethylsulfonyl fluoride) indicated that most bacterial enzymes belong to serine type protease. Our results demonstrate that aspartic proteases, which usually have high milk clotting activity, are predominantly derived from fungal strains, and therefore fungal enzymes appear to be more suitable for use in the cheese industry. Microbial enzymes studied in this research might be alternatives for rennin (chymosin) from animal source because of their low cost and stable

Antifungal Activity of Isolated Bacillus amyloliquefaciens SYBC H47 for the Biocontrol of Peach Gummosis.

PubMed

Li, Xunhang; Zhang, Yanzhou; Wei, Zhiwen; Guan, Zhengbing; Cai, Yujie; Liao, Xiangru

2016-01-01

The gummosis disease is caused by Botryosphaeria dothidea (Moug. ex. Fr) Ces. et de Not., and it is one of the most important diseases of stone fruits worldwide. The use of biocontrol as an alternative approach to synthetic chemical fungicides has aroused general concern about how to control plant diseases that are caused by phytopathogens. The aim of this study is to isolate Bacillus strains from raw honeys with the capacity to inhibit B. dothidea and to explore the mechanisms by which they could be used in the biocontrol of peach gummosis. Bacillus amyloliquefaciens SYBC H47 was isolated and identified on the basis of its physiological and biochemical characteristics and its 16S rRNA and gyrB gene sequences. The cell suspension and the cell-free supernatant of its culture showed significant antifungal activity against Aspergillus niger, Mucor racemosus, Fusarium oxysporum, Penicillium citrinum, and Candida albicans by agar-diffusion assays. The primary antifungal substances were bacillomycin L, fengycin, and surfactin, which were analyzed by HPLC LC/ESI-MS/MS. Bacillomycin L showed the best inhibitory effect against conidial germination of B. dothidea, followed by fengycin and surfactin. Surfactin had limited effects on mycelial growth, contrary to those of bacillomycin L and fengycin. However, a mixture of the three lipopeptides had a synergistic effect that disrupted the structure of the conidia and mycelia. In order to reduce the production cost, the use of waste frying peanut oil and soy oil as the sole carbon source increased the lipopeptide yield levels by approximately 17% (2.42 g/L) and 110% (4.35 g/L), respectively. In a field trial, the decreases in the infected gummosis rate (IGR) and the disease severity index (DSI) through cell suspension treatments were 20% and 57.5% (in 2014), respectively, and 40% and 57.5% (in 2015), respectively, in comparison with the control. In conclusion, B. amyloliquefaciens SYBC H47 could inhibit the germination of conidia

Microbial community dynamics during fermentation of doenjang-meju, traditional Korean fermented soybean.

PubMed

Jung, Ji Young; Lee, Se Hee; Jeon, Che Ok

2014-08-18

Bacterial and fungal community dynamics, along with viable plate counts and water content, were investigated in the exterior and interior regions of doenjang-meju, traditional Korean fermented soybean, during its fermentation process. Measurement of viable cells showed that the meju molding equipment might be an important source of bacterial cells (mostly Bacillus) during doenjang-meju fermentation, whereas fungi might be mostly derived from the fermentation environment including incubation shelves, air, and rice straws. Community analysis using rRNA-targeted pyrosequencing revealed that Bacillus among bacteria and Mucor among fungi were predominant in both the exterior and interior regions of doenjang-meju during the early fermentation period. Bacteria such as Ignatzschineria, Myroides, Enterococcus, Corynebacterium, and Clostridium and fungi such as Geotrichum, Scopulariopsis, Monascus, Fusarium, and eventually Aspergillus were mainly detected as the fermentation progressed. Bacillus, an aerobic bacterial group, was predominant in the exterior regions during the entire fermentation period, while anaerobic, facultative anaerobic, and microaerobic bacteria including Enterococcus, Lactobacillus, Clostridium, Myroides, and Ignatzschineria were much more abundant in the interior regions. Principal component analysis (PCA) also indicated that the bacterial communities in the exterior and interior regions were clearly differentiated, suggesting that aeration might be an important factor in determining the bacterial communities during doenjang-meju fermentation. However, PCA showed that fungal communities were not separated in the exterior and interior regions and Pearson's correlation coefficients showed that the major fungal taxa had significantly positive (Mucor and Geotrichum) or negative (Aspergillus) correlations with the water content during doenjang-meju fermentation, indicating that water content might be a significant factor in determining the fungal

Analysis of the effect of soil saprophytic fungi on the eggs of Baylisascaris procyonis.

PubMed

Cazapal-Monteiro, Cristiana F; Hernández, José A; Arroyo, Fabián L; Miguélez, Silvia; Romasanta, Ángel; Paz-Silva, Adolfo; Sánchez-Andrade, Rita; Arias, María S

2015-07-01

Baylisascaris procyonis is a soil-transmitted helminth mainly found in raccoons (Procyon lotor) which can also affect other domestic and sylvatic animals, as well as humans, when the eggs released in the feces of parasitized raccoons are accidentally ingested. Three assays have been conducted to assess the effect of three saprophytic fungi, Mucor circinelloides, Paecilomyces lilacinus, and Verticillium sp., on the eggs of B. procyonis. Firstly, their ovicidal effect was in vitro ascertained by placing 1 mL with 2 × 10(6) spores of each fungus in Petri plates with water-agar (2 %) and simultaneously adding 200 eggs of Baylisascaris/plate. Two in vivo probes were carried out, by spraying the fungal spores (3 mL containing about 2 × 10(6) spores/mL) on the feces of raccoons and coatis (Nasua narica) passing eggs of B. procyonis in a zoological park; the other assay consisted of evaluating the activity of the fungi after adding sand to fecal samples from raccoons. An ovicidal type 3 activity characterized by morphological damage of the eggshell with hyphal penetration, internal egg colonization, and embryo alteration was observed for all the tested fungi. In the plate assays, viability of Baylisascaris eggs reduced significantly by 53-69 % with Mucor, 45-62 % with Paecilomyces, and 52-67 % with Verticillium. A similar ovicidal effect was detected in the feces with sand. These results demonstrate the usefulness of spraying spores of M. circinelloides, Pa. lilacinus, or Verticillium sp. on the feces of animals infected by Baylisascaris to decrease the numbers of viable eggs and, thus, the risk of infection.

Assessment of the scalability of a microtiter plate system for screening of oleaginous microorganisms.

PubMed

Kosa, Gergely; Vuoristo, Kiira S; Horn, Svein Jarle; Zimmermann, Boris; Afseth, Nils Kristian; Kohler, Achim; Shapaval, Volha

2018-06-01

Recent developments in molecular biology and metabolic engineering have resulted in a large increase in the number of strains that need to be tested, positioning high-throughput screening of microorganisms as an important step in bioprocess development. Scalability is crucial for performing reliable screening of microorganisms. Most of the scalability studies from microplate screening systems to controlled stirred-tank bioreactors have been performed so far with unicellular microorganisms. We have compared cultivation of industrially relevant oleaginous filamentous fungi and microalga in a Duetz-microtiter plate system to benchtop and pre-pilot bioreactors. Maximal glucose consumption rate, biomass concentration, lipid content of the biomass, biomass, and lipid yield values showed good scalability for Mucor circinelloides (less than 20% differences) and Mortierella alpina (less than 30% differences) filamentous fungi. Maximal glucose consumption and biomass production rates were identical for Crypthecodinium cohnii in microtiter plate and benchtop bioreactor. Most likely due to shear stress sensitivity of this microalga in stirred bioreactor, biomass concentration and lipid content of biomass were significantly higher in the microtiter plate system than in the benchtop bioreactor. Still, fermentation results obtained in the Duetz-microtiter plate system for Crypthecodinium cohnii are encouraging compared to what has been reported in literature. Good reproducibility (coefficient of variation less than 15% for biomass growth, glucose consumption, lipid content, and pH) were achieved in the Duetz-microtiter plate system for Mucor circinelloides and Crypthecodinium cohnii. Mortierella alpina cultivation reproducibility might be improved with inoculation optimization. In conclusion, we have presented suitability of the Duetz-microtiter plate system for the reproducible, scalable, and cost-efficient high-throughput screening of oleaginous microorganisms.

Fungi inhabiting household environments in Riyadh, Saudi Arabia.

PubMed

Bokhary, H A; Parvez, S

1995-05-01

Fungi inhabiting household environments in the West, East and Central localities of Riyadh city were screened. The screened area included bedrooms, drawing rooms, living rooms, kitchens and bathrooms. The common genera of fungi isolated were Alternaria, Aspergillus, Cercospora, Chaetomium, Cladosporium, Curvularia, Drechslera, Embellisia, Fusarium, Mucor, Penicillium, Rhizopus, Scytalidium, Trichoderma, Torula and Ulocladium. Two uncommon genera Nodulosporium and Oidiodendron for Saudi Arabian mycoflora were also isolated. In all the localities, the highest number of fungal colonies per plate or per gram were found in the living rooms, followed by bedrooms. The number of colonies per plate or per gram was in general higher in densely populated than in less populated areas.

Kinetics of enzymatic trans-esterification of glycerides for biodiesel production.

PubMed

Calabrò, Vincenza; Ricca, Emanuele; De Paola, Maria Gabriela; Curcio, Stefano; Iorio, Gabriele

2010-08-01

In this paper, the reaction of enzymatic trans-esterification of glycerides with ethanol in a reaction medium containing hexane at a temperature of 37 degrees C has been studied. The enzyme was Lipase from Mucor miehei, immobilized on ionic exchange resin, aimed at achieving high catalytic specific surface and recovering, regenerating and reusing the biocatalyst. A kinetic analysis has been carried out to identify the reaction path; the rate equation and kinetic parameters have been also calculated. The kinetic model has been validated by comparison between predicted and experimental results. Mass transport resistances estimation was undertaken in order to verify that the kinetics found was intrinsic. Model potentialities in terms of reactors design and optimization are also shown.

The discovery of microorganisms by Robert Hooke and Antoni Van Leeuwenhoek, fellows of the Royal Society.

PubMed

Gest, Howard

2004-05-01

The existence of microscopic organisms was discovered during the period 1665-83 by two Fellows of The Royal Society, Robert Hooke and Antoni van Leeuwenhoek. In Micrographia (1665), Hooke presented the first published depiction of a microganism, the microfungus Mucor. Later, Leeuwenhoek observed and described microscopic protozoa and bacteria. These important revelations were made possible by the ingenuity of Hooke and Leeuwenhoek in fabricating and using simple microscopes that magnified objects from about 25-fold to 250-fold. After a lapse of more than 150 years, microscopy became the backbone of our understanding of the roles of microbes in the causation of infectious diseases and the recycling of chemical elements in the biosphere.

Enzymatic and Microbial Preparation of d-Xylulose from d-Xylose

PubMed Central

Chiang, Lin-Chang; Hsiao, Humg-Yu; Ueng, Pear P.; Tsao, George T.

1981-01-01

A high-d-xylulose mixture (d-xylose-d-xylulose = 33:67) was prepared from the cold ethanol extract of preisomerized d-xylose solution (d-xylose-d-xylulose = 77:23). Fusarium oxysporum f. sp. lini and Aspergillus niger were demonstrated to preferentially utilize d-xylose in the mixture of d-xylose and d-xylulose. Chromatographically pure d-xylulose was thus obtained in 90% yield. A high-d-xylulose mixture was also incubated with Rhodotorula toruloides, Klebsiella pneumoniae, Candida utilis, or Mucor rouxii.d-Xylose and d-xylulose were simultaneously consumed. When borate was added to the mixture, a d-xylulose-borate complex was formed, and it could be used to protect d-xylulose from being utilized. PMID:16345816

Isolation of Fungal Pathogens to an Edible Mushroom, Pleurotus eryngii, and Development of Specific ITS Primers

PubMed Central

Kim, Sang-Woo; Kim, Sinil; Lee, Hyun-Jun; Park, Ju-Wan

2013-01-01

Fungal pathogens have caused severe damage to the commercial production of Pleurotus eryngii, the king oyster mushroom, by reducing production yield, causing deterioration of commercial value, and shortening shelf-life. Four strains of pathogenic fungi, including Trichoderma koningiopsis DC3, Phomopsis sp. MP4, Mucor circinelloides MP5, and Cladosporium bruhnei MP6, were isolated from the bottle culture of diseased P. eryngii. A species-specific primer set was designed for each fungus from the ITS1-5.8S rDNA-ITS2 sequences. PCR using the ITS primer set yielded a unique DNA band for each fungus without any cross-reaction, proving the validity of our method in detection of mushroom fungal pathogens. PMID:24493949

Effect of Asparagus racemosus extract on transdermal delivery of carvedilol: a mechanistic study.

PubMed

Sapra, Bharti; Jain, Subheet; Tiwary, Ashok K

2009-01-01

This study was designed for investigating the effect of Asparagus racemosus (AR) extract and chitosan (CTN) in facilitating the permeation of carvedilol (CDL) across rat epidermis. Transdermal flux of carvedilol through heat-separated rat epidermis was investigated in vitro using vertical Keshary-Chien diffusion cells. Biophysical and microscopic manifestations of epidermis treated with AR extract, CTN, and AR extract-CTN mixture were investigated by using differential scanning calorimetry, transepidermal water loss, scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Biochemical estimations of cholesterol, sphingosine, and triglycerides were carried out for treated excised as well as viable rat epidermis. The antihypertensive activity of the patches in comparison with that of oral carvedilol was studied in deoxycorticosterone acetate-induced hypertensive rats. The permeation of carvedilol across excised rat epidermis was significantly higher (p < 0.05) when AR extract, CTN, or AR extract-CTN mixture was used as donor vehicle as compared to propylene glycol/ethanol (7:3) mixture. Epidermis obtained after 12 h treatment of viable rat skin with AR extract-CTN mixture showed significantly higher (p < 0.05) permeability to CDL as compared to that after treatment with AR extract or CTN alone. Further, the application of patches containing AR extract-CTN mixture resulted in sustained release of CDL which was able to control the hypertension in deoxycorticosterone acetate-induced hypertensive rats through 36 h. Estimation of micro constituents in rat epidermis revealed maximum extraction of cholesterol, sphingosine, and triglycerides after treatment with AR extract-CTN mixture. This was manifested in altered lipid and protein-specific thermotropic transitions. Further, increase in intercellular space, disordered lipid structure, and corneocyte detachment as observed in SEM and TEM suggested great potential of AR extract for use as percutaneous

The Effect of Hyperbaric Oxygen on the Growth of Mucor sp. Aspergillus fumigatus.

DTIC Science & Technology

1980-02-01

designations are for controls. 12 mamo 7<)U 4/ Figure 2. Aspergillus fumigatus Top Frame - Hyphae at day 2. Middle Frame - Plyphae with developing...conidiaphores, day 3. L~ower Frame - Hyphae with conidiophores and conidia, day 4. Time~ designations are for co~ntrols. 14 0 lOum Figure 3. Grouth curves

First report of association of Mucor circinelloides on noni (Morinda citrifolia) in Hawaii

USDA-ARS?s Scientific Manuscript database

Noni (Morinda citrifolia) is a popular medicinal plant found in tropical or subtropical regions of the world. The fruit and juice extracts have properties that are reportedly therapeutic for diabetes, high blood pressure, and certain types of cancer (1,4). In our studies on noni juice produced from ...

Phylogenetic relationship of the genus Gilbertella and related genera within the order Mucorales based on 5.8 S ribosomal DNA sequences.

PubMed

Papp, T; Acs, Klára; Nyilasi, Ildikó; Nagy, Erzsébet; Vágvölgyi, Cs

2003-01-01

The complete ITS (internal transcribed spacer) region coding the ITS1, the ITS2 and the 5.8S rDNA was amplified by polymerase chain reaction from two strains of Gilbertella persicaria, six strains in the Mucoraceae (Mucor piriformis, M. rouxii, M. circinelloides, Rhizomucor miehei, R. pusillus and R. tauricus) and four strains representing three species of the Choanephoraceae (Blakeslea trispora, Choanephora infundibulifera and Poitrasia circinans). Sequences of the amplified DNA fragments were determined and analysed. G. persicaria belongs to the monogeneric family (Gilbertellaceae), however, originally it was described as Choanephora persicaria. The goal of this study was to reveal the phylogenetic relationship among fungi belonging to Gilbertellaceae, Choanephoraceae and Mucoraceae. Our results support that the "intermediate" position of this family is between Choanephoraceae and Mucoraceae.

Primary Cutaneous Mucormycosis Caused by Rhizopus oryzae: A Case Report and Review of Literature.

PubMed

Rodríguez-Lobato, Erika; Ramírez-Hobak, Lourdes; Aquino-Matus, Jorge E; Ramírez-Hinojosa, Juan P; Lozano-Fernández, Víctor H; Xicohtencatl-Cortes, Juan; Hernández-Castro, Rigoberto; Arenas, Roberto

2017-04-01

Mucormycosis is an invasive infection caused by opportunistic fungi. Rhizopus, Lichtheimia, Mucor and Rhizomucor are the most common isolated genera. Primary cutaneous mucormycosis is usually related to traumatic injuries, but immunocompromised cases are associated with underlying conditions such as diabetes mellitus and malignancies. The treatment of choice is surgical debridement and liposomal amphotericin B. We present a 40-year-old male with fever and a painful necrotic lesion on the middle back and history of poorly controlled diabetes mellitus. Rhizopus oryzae was isolated and identified using an internal transcribed spacer regions ITS1 and ITS2. An initial good response to treatment was observed; however, 7 days later a diabetic ketoacidosis due to poor adherence to treatment caused a lethal outcome.

Effect of essential oil from fresh leaves of Ocimum gratissimum L. on mycoflora during storage of peanuts in Benin.

PubMed

Adjou, Euloge S; Kouton, Sandrine; Dahouenon-Ahoussi, Edwige; Soumanou, Mohamed M; Sohounhloue, Dominique C K

2013-02-01

The aim of this study was to evaluate the effect of essential oil from fresh leaves of Sweet Fennel (Ocimum gratissimum) on mycoflora and Aspergillus section Flavi populations in stored peanuts. Aspergillus, Fusarium and Mucor spp. were the most common genera identified from peanuts at post-harvest in Benin by using a taxonomic schemes primarily based on morphological characters of mycelium and conidia. The isolated fungi include Aspergillus niger, A. parasiticus, A. flavus, A. ochraceus, Fusarium graminearum, F. solani, F. oxysporum and Mucor spp. The most prevalent fungi recorded were A. niger (94.18 %), A. flavus (83.72 %), A. parasiticus (77.90 %), A. ochraceus (72.09 %), F. graminearum (59.30 %) and F. oxysporum (51.16 %). Antifungal assay, performed by the agar medium assay, indicated that essential oil exhibited high antifungal activity against the growth of A. flavus, A. parasiticus, A. ochraceus and F. oxysporium. The minimal inhibitory concentration (MIC) of the essential oil was found to be 7.5 μl/ml for A. flavus and A. parasiticus and 5.5 μl/ml for A. ochraceus and F. oxysporium. The minimal fungicidal concentration (MFC) was recorded to be 8.0 μl/ml for A. flavus and A. parasiticus, 6,5 μl/ml for A. ochraceus and 6.0 μl/ml for F. oxysporium. The essential oil was found to be strongly fungicidal and inhibitory to aflatoxin production. Chemical analysis by GC/MS of the components of the oil led to the identification of 31 components characterized by myrcene (6.4 %), α-thujene (8.2 %), p-cymene (17.6 %), γ-terpinene (20.0 %), and thymol (26.9 %) as major components. The essential oil of Sweet Fennel, with fungal growth and mycotoxin inhibitory properties, offers a novel approach to the management of storage, thus opening up the possibility to prevent mold contamination in stored peanuts.

[Risk factors of invasive fungal infections in patients admitted to non- hematological oncology department and pediatric intensive care unit].

PubMed

Zhao, Cheng-song; Zhao, Shun-ying; Liu, Gang; Xi-Wei, Xu

2013-08-01

To determine risk factors of invasive fungal infections (IFI) in patients admitted to non-hematological oncology department and pediatric intensive care unit (PICU), in order to improve diagnostic level of invasive fungal infections. We retrospectively assessed 85 hospitalized pediatric patients with invasive fungal infections in Beijing Children's Hospital Affiliated to Capital Medical University from Jan.2007 to Nov.2012. All the cases were either from non-hematological oncology department or the PICU.We reviewed risk factors of invasive fungal infections. Among 85 patients, 42 had invasive candida infection, 20 invasive aspergillus infection, 21 cryptococcus infection, 1 Histoplasma capsulatum infection and 1 Mucor mucedo infection.In the 42 patients with invasive candida infection, 5 were young infants, 3 had combined immunodeficiency, 1 cellular immunodeficiency, 25 secondary infection due to long term use of corticosteroids and/or combined use of more than 2 kinds of antibiotics with primary disease, 5 prior intestinal tract surgery or chronic diarrheal disease, 1 reflux gastritis.In the 20 patients with invasive aspergillosis infection, 10 patients had chronic granulomatous disease, 5 long term use of corticosteroids ≥ 1 month, 3 long term use of corticosteroids and combined use of more than 2 kinds of antibiotics, 2 had no apparent host factors.In the 21 patients with cryptococcus infection, 2 patients had used corticosteroids ≥ 1 month, 2 had immunodeficiency mainly for lack of antibodies, while others had no apparent host factors. The child with Mucor mucedo infection had diabetes mellitus. And the one with Histoplasma capsulatum infection had immunodeficiency. High risk factors for IFI in patients admitted to non-hematological oncology department and PICU are primary immunodeficiency disease and long term use of corticosteroids and/or long term combined use of more than 2 kinds of antibiotics. Besides, young infant is also a high risk factor for

New insight into microbial diversity and functions in traditional Vietnamese alcoholic fermentation.

PubMed

Thanh, Vu Nguyen; Thuy, Nguyen Thanh; Chi, Nguyen Thuy; Hien, Dinh Duc; Ha, Bui Thi Viet; Luong, Dao Thi; Ngoc, Pham Duc; Ty, Pham Van

2016-09-02

The roles of microorganisms in traditional alcoholic fermentation are often assumed based on abundance in the starter and activity in pure culture. There is a serious lack of hard evidence on the behavior and activity of individual microbial species during the actual fermentation process. In this study, microbial succession and metabolite changes during 7days of traditional Vietnamese alcoholic fermentation were monitored. Special attention was devoted to starch degradation. In total, 22 microbial species, including 6 species of filamentous fungi (Rhizopus microsporus, Rhizopus arrhizus, Mucor indicus, Mucor circinelloides, Cunninghamella elegans, Aspergillus niger), 1 yeast-like fungus (Saccharomycopsis fibuligera), 7 yeasts (Saccharomyces cerevisiae, Clavispora lusitaniae, Wickerhamomyces anomalus, Lindnera fabianii, Pichia kudriavzevii, Candida rugosa, Candida tropicalis), and 8 bacteria (Stenotrophomonas maltophilia, Lactobacillus brevis, Lactobacillus helveticus, Acinetobacter baumannii, Staphylococcus hominis, Bacillus megaterium, Enterobacter asburiae, Pediococcus pentosaceus) were identified. Despite the presence of a complex microbiota in the starter, the fermentation process is consistent and involves a limited number of functional species. Rapid change in microbial composition of fermentation mash was observed and it was correlated with ethanol content. Microbial biomass reached maximum during first 2days of solid state fermentation. Acidification of the medium took place in day 1, starch degradation in days 2, 3, 4, and alcohol accumulation from day 3. Although Sm. fibuligera dominated by cell count amongst potential starch degraders, zymography indicated that it did not produce amylase in the fermentation mash. In mixed culture with Rhizopus, amylase production by Sm. fibuligera is regulated by the moisture content of the substrate. Rhizopus was identified as the main starch degrader and S. cerevisiae as the main ethanol producer. Bacterial load was

Screening of selected single and polyherbal Ayurvedic medicines for Antibacterial and Antifungal activity

PubMed Central

Kekuda, T.R Prashith; Kavya, R; Shrungashree, R.M; Suchitra, S.V

2010-01-01

The present study deals with antimicrobial activity of ayurvedic drugs containing single herb (Amalaki Choorna and Yastimadhu Choorna) and combination of herbs (DN-90 and Asanadi Kwatha Choorna). Disc diffusion method was used to assess antibacterial activity and antifungal activity was tested using Poison food technique. Absence of bacterial growth around the discs impregnated with the aqueous extracts of drugs and reduction of fungal growth in poisoned plates indicated antimicrobial activity. Further, the results of antibacterial activity of Amalaki choorna were comparable with standard drug Streptomycin. Asanadi Kwatha Choorna inhibited bacteria to more extent than Yastimadhu choorna and DN-90. Among fungi tested, more antifungal activity was observed against Mucor sp. The antimicrobial activity of drugs tested could be due to active principles present in them. PMID:22557355

Bioactivity studies of extracts from Tridax procumbens.

PubMed

Taddei, A; Rosas-Romero, A J

2000-06-01

An updated review on the biological activity of Tridax procumbens is presented. A detailed biological screening comprised of gram-positive and gram-negative bacteria, yeasts and fungi using crude extracts of this plant was undertaken. The n-hexane extract of the flowers showed activity against Escherichia coli. The same extract of the whole aerial parts was active against Mycobacterium smegmatis, Escherichia coli, Salmonella group C and Salmonella paratyphi. The ethyl-acetate extract of the flowers was active against Bacillus cereus and Klebsiella sp. The aerial parts extract also showed activity only against Mycobacterium smegmatis and Staphylococcus aureus, while the aqueous extract showed no antimicrobial activity. None of the tested extracts was active against the yeasts, Candida albicans, Candida tropicalis and Rhodotorula rubra; or the fungi: Aspergillus flavus, Aspergillus niger, Mucor sp. and Trichophyton rubrum.

Study of cellulolytic soil fungi and two nova species and new medium

PubMed Central

Khalid, Mahmood; Yang, Wei-jun; Kishwar, Nazir; Rajput, Zahid Iqbal; Arijo, Abdullah G.

2006-01-01

This study is aimed at identifying and determining the percentage of occurrence frequency of cellulose decomposing soil fungi. The soil samples were inoculated into culture plates prepared in Sabouraud medium under sterilized conditions and incubated at 30 °C for 4 to 7 d. The identified fungal species were incubated in self-designed cellulose medium for testing their cellulolytic ability. Forty-two species, including 2 nova species, representing sixteen genera showed growth and sporulation in the cellulose medium. Most of the isolated species were from genus Aspergillus and Penicillium. Aspergillus niger and Mucor hiemalis showed highest occurrence frequency (45% and 36% respectively), as these species were collected from about 80% of soil samples. Being agar free and cheaper, the new fungal medium designed showed results equivalent to Sabouraud medium. PMID:16691640

[Effect of Trichoderma species fungi on soil micromycetes, causing infectious conifer seedling lodging in Siberian tree nurseries].

PubMed

Iakimenko, E E; Grodinitskaia, I D

2000-01-01

Soils in the tree nurseries studied were characterized by a lower species diversity of fungi than adjacent virgin soils. In particular, the relative abundances of representatives of the genera Mucor, Chaetomium, and Trichoderma in the nursery soil were two times lower than in adjacent virgin soils. On the other hand, the nursery soil exhibited greater abundances of fungi of the genus Fusarium, which are causative agents of many diseases of conifer seedlings. To appreciate the efficiency of biocontrol of the infectious diseases of conifer seedlings, we introduced several indigenous Trichoderma strains into the nursery soil and found that this affected the species composition of soil microflora considerably. Changes in the species composition of mycobiota beneficially influenced the phytosanitary state of soils and reduced the infectious lodging of conifer seedlings.

A zebrafish larval model reveals early tissue-specific innate immune responses to Mucor circinelloides.

PubMed

Voelz, Kerstin; Gratacap, Remi L; Wheeler, Robert T

2015-11-01

Mucormycosis is an emerging fungal infection that is clinically difficult to manage, with increasing incidence and extremely high mortality rates. Individuals with diabetes, suppressed immunity or traumatic injury are at increased risk of developing disease. These individuals often present with defects in phagocytic effector cell function. Research using mammalian models and phagocytic effector cell lines has attempted to decipher the importance of the innate immune system in host defence against mucormycosis. However, these model systems have not been satisfactory for direct analysis of the interaction between innate immune effector cells and infectious sporangiospores in vivo. Here, we report the first real-time in vivo analysis of the early innate immune response to mucormycete infection using a whole-animal zebrafish larval model system. We identified differential host susceptibility, dependent on the site of infection (hindbrain ventricle and swim bladder), as well as differential functions of the two major phagocyte effector cell types in response to viable and non-viable spores. Larval susceptibility to mucormycete spore infection was increased upon immunosuppressant treatment. We showed for the first time that macrophages and neutrophils were readily recruited in vivo to the site of infection in an intact host and that spore phagocytosis can be observed in real-time in vivo. While exploring innate immune effector recruitment dynamics, we discovered the formation of phagocyte clusters in response to fungal spores that potentially play a role in fungal spore dissemination. Spores failed to activate pro-inflammatory gene expression by 6 h post-infection in both infection models. After 24 h, induction of a pro-inflammatory response was observed only in hindbrain ventricle infections. Only a weak pro-inflammatory response was initiated after spore injection into the swim bladder during the same time frame. In the future, the zebrafish larva as a live whole-animal model system will contribute greatly to the study of molecular mechanisms involved in the interaction of the host innate immune system with fungal spores during mucormycosis. © 2015. Published by The Company of Biologists Ltd.

Comparison of antifungal activities of Vietnamese citrus essential oils.

PubMed

Van Hung, Pham; Chi, Pham Thi Lan; Phi, Nguyen Thi Lan

2013-03-01

Citrus essential oils (EOs) are volatile compounds from citrus peels and widely used in perfumes, cosmetics, soaps and aromatherapy. In this study, inhibition of citrus EOs extracted from Vietnamese orange (Citrus sinensis), mandarin (Citrus reticulata Blanco), pomelo (Citrus grandis Osbeck) and lime (Citrus aurantifolia Swingle) on the growth of plant pathogenic fungi, Mucor hiemalis, Penicillium expansum and Fusarium proliferatum was investigated. The EOs of the citrus peels were obtained by cold-pressing method and the antifungal activity of EOs was evaluated using the agar dilution method. The results show that the EOs had significant antifungal activity. Lime EO was the best inhibitor of M. hiemalis and F. proliferatum while pomelo EO was the most effective against P. expansum. These results indicate that citrus EOs can be used as antifungal natural products in the food, pharmaceutical and cosmetic industries.

Solitary pulmonary nodule: A rare presentation of pulmonary mucormycosis in an immunocompetent adult

PubMed Central

Sarkar, Supriya; Jash, Debraj; Maji, Arnab; Maikap, Malay Kr

2014-01-01

Pulmonary mucormycosis is a rare opportunistic infection of immunocompromised individuals. Here, we report a case of 70-year-old male, smoker presenting with high-grade fever for 2 weeks and episodes of hemoptysis. Contrast-enhanced computed tomography (CT) thorax revealed a solitary pulmonary nodule measuring 2.3 × 1.6 cm in the right upper lobe. CT guided fine needle aspiration cytology and true cut biopsy showed plenty of typical fungal hyphae consistent with the diagnosis of mucormycosis. Fungal culture confirmed the organism as mucor. Positron emission tomography-CT scan showed a non- 18 fluorodeoxy glucose avid nodule ruling out possibility of malignancy. Investigation did not reveal any evidence of immunosuppression. Patient was treated with intravenous liposomal amphotericin B for 4 weeks. Follow-up chest X-ray and CT scan after 6 weeks were normal. PMID:24669089

Specific characteristics of family 45 endoglucanases from Mucorales in the use of textiles and laundry.

PubMed

Shimonaka, Atsushi; Koga, Jinichiro; Baba, Yuko; Nishimura, Tomoko; Murashima, Koichiro; Kubota, Hidetoshi; Kono, Toshiaki

2006-04-01

We examined the characteristics of family 45 endoglucanases (glycoside hydrolases family 45; GH45) from Mucorales belonging to Zygomycota in the use of textiles and laundry. The defibrillation activities on lyocell fabric of family 45 endoglucanases from Mucorales, such as RCE1 and RCE2 from Rhizopus oryzae, MCE1 and MCE2 from Mucor circinelloides, and PCE1 from Phycomyces nitens, were much higher than those of the other family 45 endoglucanases. By contrast, family 45 endoglucanases from Mucorales were less resistant to anionic surfactant and oxidizing agent, main components in detergents, than the other family 45 endoglucanases. RCE1 consists of two distinct modules, a catalytic module and a carbohydrate-binding module family 1 (CBM1), and these common specific characteristics were considered to due to the catalytic module, but not to the CBM1.

Facial nerve paralysis: A case report of rare complication in uncontrolled diabetic patient with mucormycosis

PubMed Central

Shekar, Vandana; Sikander, Jeelani; Rangdhol, Vishwanath; Naidu, Madhulika

2015-01-01

Mucormycosis is a rare opportunistic aggressive and fatal infection caused by mucor fungus. Seven types of mucormycosis are identified based on the extension and involvement of the lesion, of which the rhino orbital mucormycosis is most common in the head and neck region. Although it is widely spread in nature, clinical cases are rare and observed only in immunocompromised patients and patients with uncontrolled diabetes mellitus. Early symptoms include fever, nasal ulceration or necrosis, periorbital edema or facial swelling, paresthesia and reduced vision. Involvement of cranial nerves although not common, facial nerve palsy is a rare finding. The infection may spread through cribriform plate to the brain resulting in extensive cerebellar infarctions. Timely diagnosis and early recognition of the signs and symptoms, correction of underlying medical disorders, and aggressive medical and surgical intervention are necessary for successful therapeutic outcome. PMID:25810669

A PLAUSIBLE ROLE FOR POLLEN-RESIDING MOLDS IN AGRICULTURAL PURPOSES.

PubMed

Belhadj, H; Harzallah, D; Dahamna, S; Ghadbane, M

2015-01-01

Pollen microbial content of 15 samples was investigated. Pollen was collected by honeybees. Total aerobic mesophilic count ranged from 3.00 to 5.48 Log CFU/g. Total mold and yeast count ranged from 2.3 to 6.99 Log CFU/g. Selected strains of isolated molds from pollen samples were characterized by conventional methods. Potent phytopathogenic and food spoilage species such as Penicillium sp., Alternaria alternata, Alternaria sp., Cladosporium werneckii, Mucor hiemalis, Rhizomucor pusillus, Aspergillus flavus, Aspergillus niger, Drechslera tritici-repentis, Verticillium albo-atrum, and Aspergillus alliaceus were recovered. Other fungal species with valuable biotechnological and plant diseases control purposes were isolated. They were characterized as Geotrichum candidum, Monilia sitophilia, and Sepedonium chrysospermum. Animal pathogenic molds were also isolated. Bee pollen may be considered as a source for a highly diverse fungal flora with different applications.

Percutaneous Management of Systemic Fungal Infection Presenting As Bilateral Renal Fungal Ball.

PubMed

Shukla, Abhishek; Shrivastava, Nitin; Singh, Chirom Amit; Nayak, Brusabhanu

2016-01-01

Background: Zygomycoses are uncommon, frequently fatal diseases caused by fungi of the class Zygomycetes. The majority of human cases are caused by Mucorales (genus-rhizopus, mucor, and absidia) fungi. Renal involvement is uncommon and urine microscopy, pottasium hydroxide mount, and fungal cultures are frequently negative. Case Presentation: A twenty-one-year-old young unmarried lady presented to our emergency department with bilateral flank pain, fever, nausea, and decreased urine output of one-month duration. She was found to have azotemia with sepsis with bilateral hydronephrosis with a left renal pelvic obstructing stone. Even after nephrostomy drainage and broad spectrum antibiotics, her condition worsened. She developed disseminated fungal infection, and timely systemic antifungal followed by bilateral nephroscopic clearance saved the patient. Conclusion: Although renal fungal infections are uncommon, a high index of suspicion and early antifungal and surgical intervention can give favorable outcomes.

Occurrence of itraconazole-tolerant micromycetes in the soil and food products.

PubMed

Piecková, E; Jesenská, Z

1999-01-01

Unexpected pathogens from the environment represent considerable risk for humans with impaired health. We examined the occurrence of itraconazole tolerant micromycetes in soil and in maize products. Five concentrations of itraconazole (2.5-12.5 micrograms/mL) selected according to known treatment schedules for human patients were incorporated into Sabouraud agar with chloramphenicol and Rose Bengal and diluted samples were inoculated onto the agar surface. After 7-d growth at 22 degrees C colonies of Alternaria sp., Aspergillus clavatus, A. glaucus group, A. flavus, A. fumigatus, A. niger group, A. ochraceus group, A. ochraceus, Chaetomium sp., Cladosporium cladosporioides, Cylindrocarpon sp., Doratomyces sp., Fusarium sp., F. moniliforme, F. oxysporum, F. solani, F. subglutinans, Marianaea elegans, Mortierella sp., Mucor sp., Myrothecium sp., Penicillium sp., Rhizopus sp., Scopulariopsis brevicaulis, Sepedonium sp., Stachybotrys chartarum, Stemphylium sp., Torula humicola and Trichoderma viride were isolated.

Mycotoxin-producing potential of fungi associated with qat (Catha edulis) leaves in Yemen.

PubMed

Mahmoud, A L

2000-01-01

Forty-four fungal species belonging to 20 genera were isolated from 30 samples of qat leaves. The most frequent genera were Aspergillus, Alternaria, Penicillium, and Cladosporium followed by Fusarium, Drechslera, Chaetomium, and Mucor. The most prevalent species in above genera were Aspergillus niger, A. flavus, A fumigatus, Alternaria alternata, Penicillium chrysogenum, P. citrinum, Cladosporium cladosporioides, and Fusarium verticillioides. From these fungi, 17 species (39%) related to 7 genera (35%) proved to be true endophytes. Eleven out of 75 isolates were mycotoxigenic. A. alternata produced alternariol and alternariol monomethyl ether whereas A. flavus produced aflatoxins B1 and B2. Ochratoxin A, sterigmatocystin, citrinin and T-2 toxin were produced by A. ochraceus, A. versicolor, P. citrinum and F. oxysporum, respectively. The presence of such toxigenic fungi associated with qat leaves is considered to be a threat to public health.

Mycobiota and mycotoxins in bee pollen collected from different areas of Slovakia.

PubMed

Kačániová, Miroslava; Juráček, Miroslav; Chlebo, Róbert; Kňazovická, Vladimíra; Kadasi-Horáková, Miriam; Kunová, Simona; Lejková, Jadža; Haščík, Peter; Mareček, Ján; Simko, Milan

2011-01-01

Contamination by microscopic fungi and mycotoxins in different bee pollen samples, which were stored under three different ways of storing as freezing, drying and UV radiation, was investigated. During spring 2009, 45 samples of bee-collected pollen were gathered from beekeepers who placed their bee colonies on monocultures of sunflower, rape and poppy fields within their flying distance. Bee pollen was collected from bees' legs by special devices placed at the entrance to hives. Samples were examined for the concentration and identification of microscopic fungi able to grow on Malt and Czapek-Dox agar and mycotoxins content [deoxynivalenol (DON), T-2 toxin (T-2), zearalenone (ZON) and total aflatoxins (AFL), fumonisins (FUM), ochratoxins (OTA)] by direct competitive enzyme-linked immunosorbent assays (ELISA). The total number of microscopic fungi in this study ranged from 2.98 ± 0.02 in frozen sunflower bee pollen to 4.06 ± 0.10 log cfu.g(-1) in sunflower bee pollen after UV radiation. In this study, 449 isolates belonging to 21 fungal species representing 9 genera were found in 45 samples of bee pollen. The total isolates were detected in frozen poppy pollen 29, rape pollen 40, sunflower pollen 80, in dried poppy pollen 12, rape pollen 36, sunflower 78, in poppy pollen after UV radiation treatment 54, rape 59 and sunflower 58. The most frequent isolates of microscopic fungi found in bee pollen samples of all prevalent species were Mucor mucedo (49 isolates), Alternaria alternata (40 isolates), Mucor hiemalis (40 isolates), Aspergillus fumigatus (33 isolates) and Cladosporium cladosporioides (31 isolates). The most frequently found isolates were detected in sunflower bee pollen frozen (80 isolates) and the lowest number of isolates was observed in poppy bee pollen dried (12 isolates). The most prevalent mycotoxin of poppy bee pollen was ZON (361.55 ± 0.26 μg.kg(-1)), in rape bee pollen T-2 toxin (265.40 ± 0.18 μg.kg(-1)) and in sunflower bee pollen T-2 toxin

Air Contamination With Fungals In Museum

NASA Astrophysics Data System (ADS)

Scarlat, Iuliana; Haiducu, Maria; Stepa, Raluca

2015-07-01

The aim of the studies was to determine the level and kind of fungal contamination of air in museum, deposits patrimony, restoration and conservation laboratories and their effects on health of workers. Microbiological air purity was measured with a SAS-100 Surface Air System impactor. The fungal contamination was observed in all 54 rooms where we made determinations. The highest levels of fungal were recorded at rooms with hygroscopic patrimony objects, eg carpets, chairs, upholstered chairs, books etc. The most species identified included under common allergens: Aspergillus, Penicillium, and Mucor. There fungal species belonging to the genus identified in this study, can trigger serious diseases museum workers, such as for example Aspergillus fumigatus, known allergies and toxic effects that may occur. In some places of the museum, occupational exposure limit values to fungi present in the air in the work environment, recommended by the specialized literature, have been overcome.

An integrated genomic and transcriptomic survey of mucormycosis-causing fungi

PubMed Central

Chibucos, Marcus C.; Soliman, Sameh; Gebremariam, Teclegiorgis; Lee, Hongkyu; Daugherty, Sean; Orvis, Joshua; Shetty, Amol C.; Crabtree, Jonathan; Hazen, Tracy H.; Etienne, Kizee A.; Kumari, Priti; O'Connor, Timothy D.; Rasko, David A.; Filler, Scott G.; Fraser, Claire M.; Lockhart, Shawn R.; Skory, Christopher D.; Ibrahim, Ashraf S.; Bruno, Vincent M.

2016-01-01

Mucormycosis is a life-threatening infection caused by Mucorales fungi. Here we sequence 30 fungal genomes, and perform transcriptomics with three representative Rhizopus and Mucor strains and with human airway epithelial cells during fungal invasion, to reveal key host and fungal determinants contributing to pathogenesis. Analysis of the host transcriptional response to Mucorales reveals platelet-derived growth factor receptor B (PDGFRB) signaling as part of a core response to divergent pathogenic fungi; inhibition of PDGFRB reduces Mucorales-induced damage to host cells. The unique presence of CotH invasins in all invasive Mucorales, and the correlation between CotH gene copy number and clinical prevalence, are consistent with an important role for these proteins in mucormycosis pathogenesis. Our work provides insight into the evolution of this medically and economically important group of fungi, and identifies several molecular pathways that might be exploited as potential therapeutic targets. PMID:27447865

Percutaneous Management of Systemic Fungal Infection Presenting As Bilateral Renal Fungal Ball

PubMed Central

Shukla, Abhishek; Shrivastava, Nitin; Singh, Chirom Amit

2016-01-01

Abstract Background: Zygomycoses are uncommon, frequently fatal diseases caused by fungi of the class Zygomycetes. The majority of human cases are caused by Mucorales (genus—rhizopus, mucor, and absidia) fungi. Renal involvement is uncommon and urine microscopy, pottasium hydroxide mount, and fungal cultures are frequently negative. Case Presentation: A twenty-one-year-old young unmarried lady presented to our emergency department with bilateral flank pain, fever, nausea, and decreased urine output of one-month duration. She was found to have azotemia with sepsis with bilateral hydronephrosis with a left renal pelvic obstructing stone. Even after nephrostomy drainage and broad spectrum antibiotics, her condition worsened. She developed disseminated fungal infection, and timely systemic antifungal followed by bilateral nephroscopic clearance saved the patient. Conclusion: Although renal fungal infections are uncommon, a high index of suspicion and early antifungal and surgical intervention can give favorable outcomes. PMID:27704055

[Indoor air studies of mould fungus contamination of homes of selected patients with bronchial asthma (with special regard to evaluation problems)].

PubMed

Senkpiel, K; Kurowski, V; Ohgke, H

1996-02-01

Investigations of indoor air of the homes of seven patients with asthma bronchiale who showed up with positive reactions following intracutaneous application of fungal allergens revealed that their places of residence were contaminated by fungal and bacterial spores. The number of colony forming units of mesophilic fungal spores of the indoor air ranged from 100 to 1000 CFU/m3 and this was much higher than the mould flora of the outdoor air determined simultaneously. The major fungi species found by the indoor investigation were: Penicillium sp. > Aspergillus sp. > Cladosporium sp., Mucor sp., Chrysonilia sp., Verticillium sp. > Geotrichum sp., Trichoderma sp. In two cases Thermoactinomyces species could be detected in the indoor air. The main cause of fungal contamination were moist building materials on room walls, insufficient air ventilation, bad maintenance of the circulating air-machines and insufficient room hygiene (e.g. biological garbage in the kitchen).

Diversity and biochemical features of culturable fungi from the coastal waters of Southern China

PubMed Central

2014-01-01

Fungi play a major role in various biogeochemical cycles of terrestrial and marine ecosystems. However, fungi in marine environments remain to be one of the most under-studied microbial groups. This study investigates the diversity of planktonic fungi from the coastal habitat off Pearl River Delta (China) using culture-dependent approach. A total of 22 fungi and 9 yeast isolates were recovered from 30 seawater and 2 sediment samples. Microscopic and ITS rRNA gene sequence analyses revealed that most of the fungi belonged to the phylum Ascomycota and Basidiomycota with a very small percentage (3%) of the subphylum Mucoromycotina of the Phylum Zygomycota. Most of these fungal isolates exhibited considerable production of extracellular enzymes, cellulase, lipase and laccase. Fungal isolates of two genera Mucor and Aspergillus sp. demonstrated pelletization capability over a wide range of pH, suggesting them as potential agents towards algae harvesting and wastewater treatment. PMID:25401065

Development of a specific polymerase chain reaction assay for the detection of Basidiobolus.

PubMed

Gómez-Muñoz, María Teresa; Fernández-Barredo, Salceda; Martínez-Díaz, Rafael Alberto; Pérez-Gracia, María Teresa; Ponce-Gordo, Francisco

2012-01-01

The etiology of chronic diarrhea is complex in humans and animals. It is always necessary to evaluate a list of differential diagnosis, including bacteria, protozoa and fungi. Basidiobolomycosis is a fungal disease reported sporadically worldwide, mainly caused by B. ranarum, a frequent organism found in soil or in the intestine and skin of lizards and frogs. It is an opportunistic pathogen that causes infections characterized by granulomatous lesions in the subcutaneous tissues as well as in the intestinal wall in humans and animals. In this work we have developed a PCR technique to differentiate Basidiobolus from other causes of intestinal disease in dogs and humans. To test the specificity of the PCR assay we included closely related organisms, common intestinal microbiota and pathogenic organisms, such as Aspergillus, Candida, Cryptosporidium, Escherichia, Giardia, Mucor, Proteus, Rhizopus and Salmonella. Pythium insidiosum, which cause clinically similar disease in dogs but require a different treatment. Only Basidiobolus was positive to the PCR assay.

Synthesis, Spectral Characterization, In-vitro Antibacterial and Antifungal Activities of Novel (2e)-Ethyl-2-(2-(2, 4-Dinitrophenyl) Hydrazono)-4-(Naphthalen-2-yl)-6-Arylcyclohex-3-Enecarboxylates

PubMed Central

Kanagarajan, V; Thanusu, J; Gopalakrishnan, M

2011-01-01

In a search for new leads towards potent antimicrobial agents, an array of novel (2E)-ethyl-2-(2-(2,4-dinitrophenyl)hydrazono)-4-(naphthalen-2-yl)-6-arylcyclohex-3-ene carboxylates 17-24 were synthesized and characterized through their melting point, elemental analysis, MS, FT-IR, one-dimensional NMR (1H, D2O exchanged 1H and 13C), two dimensional HOMOCOR and HSQC spectroscopic data. In-vitro microbiological evaluations were carried out for all the newly synthesized compounds 17-24 against clinically isolated bacterial strains namely Salmonella typhii, Klebsiellapneumoniae, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, β-Hemolytic streptococcus and Micrococcus luteusand also fungal strains namely Aspergillusflavus, Aspergillusniger, Mucor, Rhizopus and Microsporumgypseumand finally, the results of their structure activity relationship were discussed. The obtained results can be used as the key step for the building of novel chemical compounds with interesting antimicrobial profiles comparable to that of the standard drugs. PMID:24250406

Synthesis, Spectral Characterization, In-vitro Antibacterial and Antifungal Activities of Novel (2e)-Ethyl-2-(2-(2, 4-Dinitrophenyl) Hydrazono)-4-(Naphthalen-2-yl)-6-Arylcyclohex-3-Enecarboxylates.

PubMed

Kanagarajan, V; Thanusu, J; Gopalakrishnan, M

2011-01-01

In a search for new leads towards potent antimicrobial agents, an array of novel (2E)-ethyl-2-(2-(2,4-dinitrophenyl)hydrazono)-4-(naphthalen-2-yl)-6-arylcyclohex-3-ene carboxylates 17-24 were synthesized and characterized through their melting point, elemental analysis, MS, FT-IR, one-dimensional NMR ((1)H, D2O exchanged (1)H and (13)C), two dimensional HOMOCOR and HSQC spectroscopic data. In-vitro microbiological evaluations were carried out for all the newly synthesized compounds 17-24 against clinically isolated bacterial strains namely Salmonella typhii, Klebsiellapneumoniae, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, β-Hemolytic streptococcus and Micrococcus luteusand also fungal strains namely Aspergillusflavus, Aspergillusniger, Mucor, Rhizopus and Microsporumgypseumand finally, the results of their structure activity relationship were discussed. The obtained results can be used as the key step for the building of novel chemical compounds with interesting antimicrobial profiles comparable to that of the standard drugs.

Characterization of Microbial Communities in Chinese Rice Wine Collected at Yichang City and Suzhou City in China.

PubMed

Lü, Yucai; Gong, Yanli; Li, Yajie; Pan, Zejiang; Yao, Yi; Li, Ning; Guo, Jinling; Gong, Dachun; Tian, Yihong; Peng, Caiyun

2017-08-28

Two typical microbial communities from Chinese rice wine fermentation collected in Yichang city and Suzhou city in China were investigated. Both communities could ferment glutinous rice to rice wine in 2 days. The sugar and ethanol contents were 198.67 and 14.47 mg/g, respectively, for rice wine from Yichang city, and 292.50 and 12.31 mg/g, respectively, for rice wine from Suzhou city. Acetic acid and lactic acid were the most abundant organic acids. Abundant fungi and bacteria were detected in both communities by high-throughput sequencing. Saccharomycopsis fibuligera and Rhizopus oryzae were the dominant fungi in rice wine from Suzhou city, compared with R. oryzae , Wickerhamomyces anomalus, Saccharomyces cerevisiae, Mucor indicus , and Rhizopus microsporus in rice wine from Yichang city. Bacterial diversity was greater than fungal diversity in both communities. Citrobacter was the most abundant genus. Furthermore, Exiguobacterium, Aeromonas, Acinetobacter, Pseudomonas, Enterobacter, Bacillus , and Lactococcus were highly abundant in both communities.

Oxygenated N-Acyl Alanine Methyl Esters (NAMEs) from the Marine Bacterium Roseovarius tolerans EL-164.

PubMed

Bruns, Hilke; Herrmann, Jennifer; Müller, Rolf; Wang, Hui; Wagner Döbler, Irene; Schulz, Stefan

2018-01-26

The marine bacterium Roseovarius tolerans EL-164 (Rhodobacteraceae) can produce unique N-acylalanine methyl esters (NAMEs) besides strucutrally related N-acylhomoserine lactones (AHLs), bacterial signaling compounds widespread in the Rhodobacteraceae. The structures of two unprecedented NAMEs carrying a rare terminally oxidized acyl chain are reported here. The compounds (Z)-N-16-hydroxyhexadec-9-enoyl-l-alanine methyl ester (Z9-16-OH-C16:1-NAME, 3) and (Z)-N-15-carboxypentadec-9-enoyl-l-alanine methyl ester (16COOH-C16:1-NAME, 4) were isolated, and the structures were determined by NMR and MS experiments. Both compounds were synthesized to prove assignments and to test their biological activity. Finally, non-natural, structurally related Z9-3-OH-C16:1-NAME (18) was synthesized to investigate the mass spectroscopy of structurally related NAMEs. Compound 3 showed moderate antibacterial activity against microorganisms such as Bacillus, Streptococcus, Micrococcus, or Mucor strains. In contrast to AHLs, quorum-sensing or quorum-quenching activity was not observed.

The diversity of eukaryotic microbiota in the traditional Slovak sheep cheese--bryndza.

PubMed

Laurencík, M; Sulo, P; Sláviková, E; Piecková, E; Seman, M; Ebringer, L

2008-09-30

We investigated the occurrence and diversity of yeasts and filamentous fungi in bryndza an artisanal Slovak soft spreadable cheese prepared from raw sheep milk or from a mixture of sheep and cow milk. Samples collected during four months of the summer production period from two locations (northern and southern parts of central Slovakia) contained 10(5)-10(7) (cfu) yeasts and about 10(2) (cfu) of mold per gram of wet weight. Further characterization by conventional taxonomy and sequence comparison of D1/D2 region from 26S rRNA gene revealed Mucor circinelloides v. Tieghem as the predominant filamentous fungus. A novel Geotrichum sp. together with Kluyveromyces (K. lactis/K. marxianus) was identified as the most abundant yeast species. Occasionally other yeasts, such as Candida inconspicua, Candida silvae, Pichia fermentans and Trichosporon domesticum were found. Conventional taxonomy readily identified isolates to the genus level, but DNA sequence comparison was capable of discriminating them at the species level.

Invasive pulmonary Aspergillosis in organ transplants--Focus on lung transplants.

PubMed

Geltner, Christian; Lass-Flörl, Cornelia

2016-03-01

Infections with filamentous fungi are common in transplant recipients. The risk for aspergillosis and other invasive pulmonary mycosis (IPM) is high in patients undergoing stem cell and lung transplantations. The mortality rates range from 20% to 60% and depend on a number of risk factors. The typical manifestations of IPM are lung infiltrates, consolidations, and fungal tracheobronchitis. The most common infectious agent is Aspergillus fumigatus. Infections caused by non-Aspergillus molds are more frequent for various reasons. The species distribution of non-Aspergillus molds varies in different locations. Furthermore, infections caused by Mucor and Penicillium are increasing, as are infections caused by species resistant to azoles and amphotericin B. Most centers use antifungal prophylaxis with inhaled amphotericin B or oral azoles. Early diagnosis and therapy is crucial. Reliable information on the local microbiological spectrum is a prerequisite for the effective treatment of molds with primary or secondary resistance to antimycotic drugs. Copyright © 2015 The Japanese Respiratory Society. Published by Elsevier B.V. All rights reserved.

Biocatalyst mediated production of 6β,11α-dihydroxy derivatives of 4-ene-3-one steroids.

PubMed

Kolet, Swati P; Niloferjahan, Siddiqui; Haldar, Saikat; Gonnade, Rajesh; Thulasiram, Hirekodathakallu V

2013-11-01

Biotransformation of steroids with 4-ene-3-one functionality such as progesterone (I), testosterone (II), 17α-methyltestosterone (III), 4-androstene-3,17-dione (IV) and 19-nortestosterone (V) were studied by using a fungal system belonging to the genera of Mucor (M881). The fungal system efficiently and quantitatively converted these steroids in regio- and stereo-selective manner into corresponding 6β,11α-dihydroxy compounds. Time course experiments suggested that the transformation was initiated by hydroxylation at 6β- or 11α-(10β-hydroxy in case of V) to form monohydroxy derivatives which upon prolonged incubation were converted into corresponding 6β,11α-dihydroxy derivatives. The fermentation studies carried out using 5L table-top fermentor with substrates (I and II) clearly indicates that 6β,11α-dihydroxy derivatives of steroids with 4-ene-3-one functionality can be produced in large scale by using M881. Copyright © 2013 Elsevier Ltd. All rights reserved.

Biogenic Silver Nanoparticles by Gelidiella acerosa Extract and their Antifungal Effects

PubMed Central

Vivek, Marimuthu; Kumar, Palanisamy Senthil; Steffi, Sesurajan; Sudha, Sellappa

2011-01-01

The synthesis, characterization and application of biologically synthesized nanomaterials are an important aspect in nanotechnology. The present study deals with the synthesis of silver nanoparticles (Ag-NPs) using the aqueous extract of red seaweed Gelidiella acerosa as the reducing agent to study the antifungal activity. The formation of Ag-NPs was confirmed by UV-Visible Spectroscopy, X-Ray Diffraction (XRD) pattern, Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). The synthesized Ag-NPs was predominately spherical in shape and polydispersed. Fourier Transform Infra-Red (FT-IR) spectroscopy analysis showed that the synthesized nano-Ag was capped with bimolecular compounds which are responsible for reduction of silver ions. The antifungal effects of these nanoparticles were studied against Humicola insolens (MTCC 4520), Fusarium dimerum (MTCC 6583), Mucor indicus (MTCC 3318) and Trichoderma reesei (MTCC 3929). The present study indicates that Ag-NPs have considerable antifungal activity in comparison with standard antifungal drug, and hence further investigation for clinical applications is necessary. PMID:23408653

Zygomycetes From Herbivore Dung in the Ecological Reserve of Dois IrmÃOs, Northeast Brazil

PubMed Central

de Azevedo Santiago, André Luiz Cabral Monteiro; Botelho Trufem, Sandra Farto; Malosso, Elaine; dos Santos, Paulo Jorge Parreira; de Queiroz Cavalcanti, Maria Auxiliadora

2011-01-01

Thirty-eight taxa of Zygomycetes distributed in 15 genera were recorded from tapir (Tapirus terrestris), camel (Camelus bactrianus), horse (Equus caballus), deer (Cervus elaphus), agouti (Dasyprocta aguti), donkey (Equus asinus), llama (Llama glama) and waterbuck (Kobus ellipsiprymnus) dung collected at the Reserva Ecológica de Dois Irmãos located in Recife, State of Pernambuco, Northeast Brazil. The samples were collected on a monthly basis from June 2005 to May 2006, taken to the laboratory and incubated in moist chambers. Higher number of taxa was observed in the excrements of tapir, followed by deer and donkey. The highest number of species was detected for Mucor, followed by Pilobolus. Statistical analyses showed significant differences in richness of Zygomycetes taxa between the herbivore dung types. Differences of species composition, however, were weak. Seasonality influenced the Zygomycetes species composition but not its richness. Variations in taxa composition between ruminants and non-ruminants dung were non significant. PMID:24031609

Zygomycetes From Herbivore Dung in the Ecological Reserve of Dois IrmÃOs, Northeast Brazil.

PubMed

de Azevedo Santiago, André Luiz Cabral Monteiro; Botelho Trufem, Sandra Farto; Malosso, Elaine; Dos Santos, Paulo Jorge Parreira; de Queiroz Cavalcanti, Maria Auxiliadora

2011-01-01

Thirty-eight taxa of Zygomycetes distributed in 15 genera were recorded from tapir (Tapirus terrestris), camel (Camelus bactrianus), horse (Equus caballus), deer (Cervus elaphus), agouti (Dasyprocta aguti), donkey (Equus asinus), llama (Llama glama) and waterbuck (Kobus ellipsiprymnus) dung collected at the Reserva Ecológica de Dois Irmãos located in Recife, State of Pernambuco, Northeast Brazil. The samples were collected on a monthly basis from June 2005 to May 2006, taken to the laboratory and incubated in moist chambers. Higher number of taxa was observed in the excrements of tapir, followed by deer and donkey. The highest number of species was detected for Mucor, followed by Pilobolus. Statistical analyses showed significant differences in richness of Zygomycetes taxa between the herbivore dung types. Differences of species composition, however, were weak. Seasonality influenced the Zygomycetes species composition but not its richness. Variations in taxa composition between ruminants and non-ruminants dung were non significant.

A multi-criteria analysis approach for ranking and selection of microorganisms for the production of oils for biodiesel production.

PubMed

Ahmad, Farah B; Zhang, Zhanying; Doherty, William O S; O'Hara, Ian M

2015-08-01

Oleaginous microorganisms have potential to be used to produce oils as alternative feedstock for biodiesel production. Microalgae (Chlorella protothecoides and Chlorella zofingiensis), yeasts (Cryptococcus albidus and Rhodotorula mucilaginosa), and fungi (Aspergillus oryzae and Mucor plumbeus) were investigated for their ability to produce oil from glucose, xylose and glycerol. Multi-criteria analysis (MCA) using analytic hierarchy process (AHP) and preference ranking organization method for the enrichment of evaluations (PROMETHEE) with graphical analysis for interactive aid (GAIA), was used to rank and select the preferred microorganisms for oil production for biodiesel application. This was based on a number of criteria viz., oil concentration, content, production rate and yield, substrate consumption rate, fatty acids composition, biomass harvesting and nutrient costs. PROMETHEE selected A. oryzae, M. plumbeus and R. mucilaginosa as the most prospective species for oil production. However, further analysis by GAIA Webs identified A. oryzae and M. plumbeus as the best performing microorganisms. Copyright © 2015 Elsevier Ltd. All rights reserved.

Assessment of microbial contamination within working environments of different types of composting plants.

PubMed

Gutarowska, Beata; Skóra, Justyna; Stępień, Łukasz; Szponar, Bogumiła; Otlewska, Anna; Pielech-Przybylska, Katarzyna

2015-04-01

The objective of the study was to determine the degree of microbiological contamination, type of microflora, bioaerosol particle size distribution, and concentration of endotoxins in dust in different types of composting plants. In addition, this study provides a list of indicator microorganisms that pose a biological threat in composting facilities, based on their prevalence within the workplace, source of isolation, and health hazards. We undertook microbiological analysis of the air, work surfaces, and compost, and assessed the particle size distribution of bioaerosols using a six-stage Andersen sampler. Endotoxins were determined using gas chromatography-mass spectrometry (GC-MS). Microbial identification was undertaken both microscopically and using biochemical tests. The predominant bacterial and fungal species were identified using 16S rRNA and ITS1/2 analysis, respectively. The number of mesophilic microorganisms in composting plants amounted to 6.9×10(2)-2.5×10(4) CFU/m3 in the air, 2.9×10(2)-3.3×10(3) CFU/100 cm2 on surfaces, and 2.2×10(5)-2.4×10(7) CFU/g in compost. Qualitative analysis revealed 75 microbial strains in composting plants, with filamentous fungi being the largest group of microorganisms, accounting for as many as 38 isolates. The total amount of endotoxins was 0.0062-0.0140 nmol/mg of dust. The dust fraction with aerodynamic particle diameter of 0.65-1.1 μm accounted for 28-39% of bacterial aerosols and 4-13% of fungal aerosols. We propose the following strains as indicators of harmful biological agent contamination: Bacillus cereus, Aspergillus fumigatus, Cladosporium cladosporioides, C. herbarum, Mucor hiemalis, and Rhizopus oryzae for both types of composting plants, and Bacillus pumilus, Mucor fragilis, Penicillium svalbardense, and P. crustosum for green waste composting plants. The biological hazards posed within these plants are due to the presence of potentially pathogenic microorganisms and the inhalation of respirable

Purification of a fibrinolytic protease from Mucor subtilissimus UCP 1262 by aqueous two-phase systems (PEG/sulfate).

PubMed

Nascimento, Thiago Pajeú; Sales, Amanda Emmanuelle; Porto, Camila Souza; Brandão, Romero Marcos Pedrosa; de Campos-Takaki, Galba Maria; Teixeira, José Antônio Couto; Porto, Tatiana Souza; Porto, Ana Lúcia Figueiredo; Converti, Attilio

2016-07-01

A fibrinolytic protease from M. subtilissimus UCP 1262 was recovered and partially purified by polyethylene glycol (PEG)/sodium sulfate aqueous two-phase systems (ATPS). The simultaneous influence of PEG molar mass, PEG concentration and sulfate concentration on the enzyme recovery was first investigated using a 2(3) full factorial design, and the Response Surface Methodology used to identify the optimum conditions for enzyme extraction by ATPS. Once the best PEG molar mass for the process had been selected (6000g/mol), a two-factor central composite rotary design was applied to better evaluate the effects of the other two independent variables. The fibrinolytic enzyme was shown to preferentially partition to the bottom phase with a partition coefficient (K) ranging from 0.2 to 0.7. The best results in terms of enzyme purification were obtained with the system formed by 30.0% (w/w) PEG 6000g/mol and 13.2% (w/w) sodium sulfate, which ensured a purification factor of 10.0, K of 0.2 and activity yield of 102.0%. SDS-PAGE and fibrin zymography showed that the purified protease has a molecular mass of 97kDa and an apparent isoelectric point of 5.4. When submitted to assays with different substrates and inhibitors, it showed selectivity for succinyl-l-ala-ala-pro-l-phenylalanine-p-nitroanilide and was almost completely inhibited by phenylmethylsulfonyl fluoride, behaving as a chymotrypsin-like protease. At the optimum temperature of 37°C, the enzyme residual activity was 94 and 68% of the initial one after 120 and 150min of incubation, respectively. This study demonstrated that M. subtilissimus protease has potent fibrinolytic activity compared with similar enzymes produced by solid-state fermentation, therefore it may be used as an agent for the prevention and therapy of thrombosis. Furthermore, it appears to have the advantages of low cost production and simple purification. Copyright © 2016 Elsevier B.V. All rights reserved.

Lipid formation and γ-linolenic acid production by Mucor circinelloides and Rhizopus sp., grown on vegetable oil

PubMed Central

Tauk-Tornisielo, Sâmia M.; Arasato, Luciana S.; de Almeida, Alex F.; Govone, José S.; Malagutti, Eleni N.

2009-01-01

The fungi strains were tested in Bioscreen automated system to select the best nutritional source. Following, shaking submserse cultures were studied in media containing sole carbon or nitrogen source. The growth of these strains improved in media containing vegetable oil, with high concentration of lipids. The high concentration of γ-linolenic acid was obtained with M. circinelloides in culture containing sesame oil. PMID:24031370

Cloning and expression of two chitin deacetylase genes of Saccharomyces cerevisiae.

PubMed

Mishra, C; Semino, C E; McCreath, K J; de la Vega, H; Jones, B J; Specht, C A; Robbins, P W

1997-03-30

Chitin deacetylase (EC 3.5.1.41), which hydrolyses the N-acetamido groups of N-acetyl-D-glucosamine residues in chitin, has been demonstrated in crude extracts from sporulating Saccharomyces cerevisiae. Two S. cerevisiae open reading frames (ORFs), identified by the Yeast Genome Project, have protein sequence homology to a chitin deacetylase from Mucor rouxii. Northern blot hybridizations show each ORF was transcribed in diploid cells after transfer to sporulation medium and prior to formation of asci. Each ORF was cloned in a vector under transcriptional control of the GAL 1, 10 promoter and introduced back into haploid strains of S. cerevisiae. Chitin deacetylase activity was detected by in vitro assays from vegetative cells grown in galactose. Chemical analysis of these cells also demonstrated the synthesis of chitosam in vivo. Both recombinant chitin deacetylases showed similar qualitative and quantitative activities toward chitooligosaccharides in vitro. A diploid strain deleted to both ORFs, when sporulated, did not show deacetylase activity. The mutant spores were hypersensitive to lytic enzymes (Glusulase or Zymolyase).

Mucorales species activation of a serum leukotactic factor.

PubMed Central

Marx, R S; Forsyth, K R; Hentz, S K

1982-01-01

Previous studies have suggested that the focal accumulation of phagocytic leukocytes is an important feature of the host response in mucormycosis. To ascertain the basis for this influx of inflammatory cells, we evaluated the effect of members of the order Mucorales, including species from the genera Rhizopus, Absidia, and Mucor, on the chemotactic activity of normal human serum for neutrophils and monocytes. Both hyphae and spores produced concentration-dependent chemotaxigenesis in serum to a maximum level equivalent to that produced by zymosan activation of serum. Chemotactic activity was similar for live and heat-killed hyphae. No leukotactic activity was demonstrated in the absence of serum. The pretreatment of serum with anti-C3 antibody, heating at 56 degrees C, or 0.01 M EDTA abolished the activity. The pretreatment of serum with 0.01 M ethylene glycol-bis(beta-aminoethyl ether)-N,N-tetraacetic acid did not abolish the activity. These data provide evidence that the leukotactic activity of Mucorales species is generated through the alternative complement pathway. PMID:6759409

Antifungal activity of peppermint and sweet basil essential oils and their major aroma constituents on some plant pathogenic fungi from the vapor phase.

PubMed

Edris, Amr E; Farrag, Eman S

2003-04-01

The vapors of peppermint oil and two of its major constituents (menthol and menthone), and sweet basil oil and two of its major constituents (linalool and eugenol), were tested against Sclerotinia sclerotiorum (Lib.), Rhizopus stolonifer (Ehrenb. exFr.) Vuill and Mucor sp. (Fisher) in a closed system. These fungi cause deterioration and heavy decay of peach fruit during marketing, shipping and storage. The essential oils, their major individual aroma constituents and blends of the major individual constituents at different ratios inhibited the growth of the fungi in a dose-dependent manner. Menthol was found to be the individual aroma constituent responsible for the antifungal properties of peppermint essential oil, while menthone alone did not show any effect at all doses. In the case of basil oil, linalool alone showed a moderate antifungal activity while eugenol showed no activity at all. Mixing the two components in a ratio similar to their concentrations in the original oil was found to enhance the antifungal properties of basil oil indicating a synergistic effect.

Isolation and Identification of Mushroom Pathogens from Agrocybe aegerita

PubMed Central

Choi, Jang-Nam; Sharma, Praveen K.; Lee, Wang-Hyu

2010-01-01

Agrocybe aegerita is an important mushroom cultivated in Korea, with good feel and a peculiar fragrance. A. aegerita can be cultivated throughout the year using culture bottles but is more susceptible to contamination than other mushrooms. Twenty-two pathogens were isolated from the fruiting bodies and compost of A. aegerita, and seven isolates were isolated from Pleurotus ostreatus to compare with the A. aegerita isolates, collected from Gimje, Iksan, Gunsan of Chonbuk, and Chilgok of Gyeongbuk Province in 2009. These isolates were identified based on morphological and molecular characteristics. Of the 29 isolates, 26 were identified as Trichoderma spp. and the remaining three were Aspergillus spp., Mucor spp., and Penicillium spp. A phylogenetic analysis revealed that the 26 isolates of Trichoderma were divided into four taxa, namely T. harzianum, T. pleuroticola, T. longibrachiatum, and T. atroviride. Among the Trichoderma spp., 16 isolates (55.2%) were identified as T. harzianum, six as T. pleuroticola (20.7%), two as T. longibrachiatum, and the remaining two were T. atroviride. PMID:23956671

Algological and Mycological Characterization of Soils under Pine and Birch Forests in the Pasvik Reserve

NASA Astrophysics Data System (ADS)

Korneikova, M. V.; Redkina, V. V.; Shalygina, R. R.

2018-02-01

The structure of algological and mycological complexes in Al-Fe-humus podzols (Albic Podzols) under pine and birch forests of the Pasvik Reserve is characterized. The number of micromycetes is higher in more acid soils of the pine forest, while the species diversity is greater under the birch forest. The genus Penicillium includes the largest number of species. The greatest abundance and occurrence frequency are typical for Penicillium spinulosum, P. glabrum, and Trichoderma viride in pine forest and for Umbelopsis isabellina, Mucor sp., Mortierella alpina, P. glabrum, Aspergillus ustus, Trichoderma viride, and T. koningii in birch forest. Cyanobacteria-algal cenoses of the investigated soils are predominated by green algae. Soils under birch forest are distinguished by a greater diversity of algal groups due to the presence of diatoms and xanthophytes. Species of frequent occurrence are represented by Pseudococcomyxa simplex and Parietochloris alveolaris in soils of the pine forest and by Tetracystis cf. aplanospora, Halochlorella rubescens, Pseudococcomyxa simplex, Fottea stichococcoides, Klebsormidium flaccidum, Hantzschia amphioxys, Microcoleus vaginatus, and Aphanocapsa sp. in soils under birch forest

Isolation of Ovicidal Fungi from Fecal Samples of Captive Animals Maintained in a Zoological Park.

PubMed

Hernández, José A; Vázquez-Ruiz, Rosa A; Cazapal-Monteiro, Cristiana F; Valderrábano, Esther; Arroyo, Fabián L; Francisco, Iván; Miguélez, Silvia; Sánchez-Andrade, Rita; Paz-Silva, Adolfo; Arias, María S

2017-06-02

Abstract : There are certain saprophytic fungi in the soil able to develop an antagonistic effect against eggs of parasites. Some of these fungal species are ingested by animals during grazing, and survive in their feces after passing through the digestive tract. To identify and isolate ovicidal fungi in the feces of wild captive animals, a total of 60 fecal samples were taken from different wild animals kept captive in the Marcelle Natureza Zoological Park (Lugo, Spain). After the serial culture of the feces onto Petri dishes with different media, their parasicitide activity was assayed against eggs of trematodes ( Calicophoron daubneyi ) and ascarids ( Parascaris equorum ). Seven fungal genera were identified in the feces. Isolates from Fusarium , Lecanicillium , Mucor , Trichoderma , and Verticillium showed an ovicidal effect classified as type 3, because of their ability to adhere to the eggshell, penetrate, and damage permanently the inner embryo. Penicillium and Gliocladium developed a type 1 effect (hyphae attach to the eggshell but morphological damage was not provoked). These results provide very interesting and useful information about fungi susceptible for being used in biological control procedures against parasites.

Moisture Sorption Behaviour and Mould Ecology of Trade Garri Sold in South Eastern Nigeria

PubMed Central

Samuel, Tochukwu; Ugwuanyi, J. Obeta

2014-01-01

Garri is a creamy white or yellow starchy grit produced by roasting to gelatinization and dryness of peeled, washed, mashed, and fermented dewatered cassava roots. It is the most important product of cassava in West and Central Africa. Mean moisture content of yellow and white garri was 11.11% and 10.81% within 24 hrs of sampling from the market, increasing to 17.27% and 16.14%, respectively, following 3 months of storage at room temperature. The water activity of samples varied from initial 0.587 to 0.934 following storage. Moisture sorption isotherms, determined by static gravimetric techniques at 20° and 30°C, showed temperature dependent BET Sigmoidal type II behaviour typical of carbohydrate rich foods but modulated very slightly by the content of palm oil. Equilibrium moisture content decreased with increase in temperature at constant water activity. A total of 10 fungal species belonging to the genera Mucor, Penicillium, Cephalosporium, Aspergillus, Scopulariopsis, Rhizopus, and Paecilomyces were identified, with range increasing with water activity of samples. PMID:26904621

Simultaneous enrichment of cereals with polyunsaturated fatty acids and pigments by fungal solid state fermentations.

PubMed

Čertík, Milan; Adamechová, Zuzana; Guothová, Lucia

2013-10-20

Four Mucor strains were tested for their ability to grow on four cereal substrates and enriched them with gamma-linolenic acid (GLA) and β-carotene. M. circinelloides CCF-2617 as the best producer accumulated of both GLA and β-carotene in high amounts during utilization of rye bran/spent malt grains (3:1). The first growth phase was characterized by rapid GLA biosynthesis, while distinct β-carotene formation was found in the stationary fungal growth. Therefore various cultivation conditions were tested in order to optimize the yield of either GLA or β-carotene. The fungus grown on cereal substrate supplemented with glucose produced maximal 8.5 mg β-carotene and 12.1 g GLA in 1 kg fermented substrate, respectively. On the other hand, the highest amount of GLA in the fermented substrate (24.2 g/kg) was achieved when 30% of sunflower oil was employed to the substrate. Interestingly, β-carotene biosynthesis was completely inhibited when either whey or linseed oil were added to the substrate. Copyright © 2013 Elsevier B.V. All rights reserved.

Isolation and characterization of bioactive fungi from shark Carcharodon carcharias' gill with biopharmaceutical prospects

NASA Astrophysics Data System (ADS)

Zhang, Yi; Han, Jinyuan; Feng, Yan; Mu, Jun; Bao, Haiyan; Kulik, Andreas; Grond, Stephanie

2016-01-01

Until recently, little was known about the fungi found in shark gills and their biomedicinal potential. In this article, we described the isolation, bioactivity, diversity, and secondary metabolites of bioactive fungi from the gill of a shark ( Carcharodon carcharias). A total of 115 isolates were obtained and grown in 12 culture media. Fifty-eight of these isolates demonstrated significant activity in four antimicrobial, pesticidal, and cytotoxic bioassay models. Four randomly selected bioactive isolates inhibited human cancer cell proliferation during re-screening. These active isolates were segregated into 6 genera using the internal transcribed spacer-large subunit (ITS-LSU) rDNA-sequence BLAST comparison. Four genera, Penicillium, Aspergillus, Mucor, and Chaetomium were the dominant taxa. A phylogenic tree illustrated their intergenera and intragenera genetic diversity. HPLC-DAD-HRMS analysis and subsequent database searching revealed that nine representative strains produced diverse bioactive compound profiles. These results detail the broad range of bioactive fungi found in a shark's gills, revealing their biopharmaceutical potential. To the best of our knowledge, this is the first study characterizing shark gill fungi and their bioactivity.

Transglycosidase-like activity of Mucor hiemalis endoglycosidase mutants enabling the synthesis of glycoconjugates using a natural glycan donor.

PubMed

Sakaguchi, Kouta; Katoh, Toshihiko; Yamamoto, Kenji

2016-11-01

Glycan conversion of glycoprotein via the transglycosylation activity of endo-β-N-acetylglucosaminidase is a promising chemoenzymatic technology for the production of glycoproteins including bio-medicines with a homogeneous glycoform. Although Endo-M is a key enzyme in this process, its product undergoes rehydrolysis, which leads to a lower yield, and limits the practical application of this enzyme. We developed several Endo-M mutant enzymes including N175Q with glycosynthase-like activity and/or transglycosidase-like activity. We found that the Endo-M N175H mutant showed glycosynthase-like activity comparable to N175Q as well as transglycosidase-like activity superior to N175Q. Using a natural sialylglycopeptide as a donor substrate, N175H readily transferred the sialo-glycan onto an N-acetylglucosamine residue attached to bovine ribonuclease B (RNase B), yielding a nonnative sialoglycosylated RNase B. These results demonstrate that use of Endo-M N175H is an alternative glycoengineering technique, which provides a relatively high yield of transglycosylation product and avoids the laborious synthesis of a sugar oxazoline as a donor substrate. © 2015 International Union of Biochemistry and Molecular Biology, Inc.

Chlorine Dioxide for Reduction of Postharvest Pathogen Inoculum during Handling of Tree Fruits

PubMed Central

Roberts, Rodney G.; Reymond, Stephen T.

1994-01-01

Alternatives to hypochlorous acid and fungicides are needed for treatment of fruit and fruit-handling facilities. Chlorine dioxide was evaluated and found effective against common postharvest decay fungi and against filamentous fungi occurring on fruit packinghouse surfaces. In vitro tests with conidial or sporangiospore suspensions of Botrytis cinerea, Penicillium expansum, Mucor piriformis, and Cryptosporiopsis perennans demonstrated >99% spore mortality within 1 min when the fungi were exposed to aqueous chlorine dioxide at 3 or 5 μg · ml-1. Longer exposure times were necessary to achieve similar spore mortalities with 1 μg · ml-1. Of the fungi tested, B. cinerea and P. expansum were the least sensitive to ClO2. In comparison with the number recovered from untreated control areas, the number of filamentous fungi recovered was significantly lower in swipe tests from hard surfaces such as belts and pads in a commercial apple and pear packinghouse after treatment of surfaces with a 14.0- to 18.0-μg · ml-1 ClO2 foam formulation. Chlorine dioxide has desirable properties as a sanitizing agent for postharvest decay management when residues of postharvest fungicides are not desired or allowed. PMID:16349354

Lipase-catalyzed synthesis of palmitanilide: Kinetic model and antimicrobial activity study.

PubMed

Liu, Kuan-Miao; Liu, Kuan-Ju

2016-01-01

Enzymatic syntheses of fatty acid anilides are important owing to their wide range of industrial applications in detergents, shampoo, cosmetics, and surfactant formulations. The amidation reaction of Mucor miehei lipase Lipozyme IM20 was investigated for direct amidation of triacylglycerol in organic solvents. The process parameters (reaction temperature, substrate molar ratio, enzyme amount) were optimized to achieve the highest yield of anilide. The maximum yield of palmitanilide (88.9%) was achieved after 24 h of reaction at 40 °C at an enzyme concentration of 1.4% (70 mg). Kinetics of lipase-catalyzed amidation of aniline with tripalmitin has been investigated. The reaction rate could be described in terms of the Michaelis-Menten equation with a Ping-Pong Bi-Bi mechanism and competitive inhibition by both the substrates. The kinetic constants were estimated by using non-linear regression method using enzyme kinetic modules. The enzyme operational stability study showed that Lipozyme IM20 retained 38.1% of the initial activity for the synthesis of palmitanilide (even after repeated use for 48 h). Palmitanilide, a fatty acid amide, exhibited potent antimicrobial activity toward Bacillus cereus. Copyright © 2015 Elsevier Inc. All rights reserved.

Inhibitory effect of gamma radiation and Nigella sativa seeds oil on growth, spore germination and toxin production of fungi

NASA Astrophysics Data System (ADS)

Zeinab, E. M. EL-Bazza; Hala, A. Farrag; Mohie, E. D. Z. EL-Fouly; Seham, Y. M. EL-Tablawy

2001-02-01

Twenty samples of Nigella sativa seeds (Black cumin) were purchased from different localities in Egypt. The mold viable count ranged from 1.7×10 1 to 9.8×10 3 c.f.u. Sixty six molds were isolated belonging to six genera Aspergillus, Penicillium, Rhizopus, Mucor, Alternaria and Fusarium. Exposure of seeds samples to different radiation doses showed that a dose level of 6.0 kGy could be considered as a sufficient dose for decontamination of the tested samples. Seven radioresistant isolates were identified as Rhizopus oryzae, Rhizopus stolonifer, Penicillium chrysogenum and Penicillium corylophillum. All the herb samples were found to be free from aflatoxins B 1, B 2, G 1, G 2 and ochratoxin A. One mold isolate was identified as Aspergillus flavus could produce aflatoxin B 1 and G 1. None of the isolated radioresistant strains could produce mycotoxins. The water activities of seeds were slightly decreased by the storage time and the seeds needed to be stored at relative humidity not more than 85%. The addition of extract volatile and fixed oil from tested seeds to the medium stimulated the growth of isolated Aspergillus sp.

Comparative Analysis of Pathogenic Organisms in Cockroaches from Different Community Settings in Edo State, Nigeria

PubMed Central

Orue, Philip Ogbeide; Iyamu, Mercy Itohan; Ehiaghe, Joy Imuetiyan; Isaac, Osesojie

2014-01-01

Cockroaches are abundant in Nigeria and are seen to harbour an array of pathogens. Environmental and sanitary conditions associated with demographic/socio-economic settings of an area could contribute to the prevalence of disease pathogens in cockroaches. A total of 246 cockroaches (Periplaneta americana) in urban (Benin, n=91), semi-urban (Ekpoma, n=75) and rural (Emuhi, n=70) settings in Edo State, Nigeria were collected within and around households. The external body surfaces and alimentary canal of these cockroaches were screened for bacterial, fungal, and parasitological infections. Bacillus sp. and Escherichia coli were the most common bacteria in cockroaches. However, Enterococcus faecalis could not be isolated in cockroaches trapped from Ekpoma and Emuhi. Aspergillus niger was the most prevalent fungus in Benin and Ekpoma, while Mucor sp. was predominant in Emuhi. Parasitological investigations revealed the preponderance of Ascaris lumbricoides in Benin and Emuhi, while Trichuris trichura was the most predominant in Ekpoma. The prevalence and burden of infection in cockroaches is likely to be a reflection of the sanitary conditions of these areas. Also, cockroaches in these areas making incursions in homes may increase the risk of human infections with these disease agents. PMID:24850961

Hydrolysis of virgin coconut oil using immobilized lipase in a batch reactor.

PubMed

Chua, Lee Suan; Alitabarimansor, Meisam; Lee, Chew Tin; Mat, Ramli

2012-01-01

Hydrolysis of virgin coconut oil (VCO) had been carried out by using an immobilised lipase from Mucor miehei (Lipozyme) in a water-jacketed batch reactor. The kinetic of the hydrolysis was investigated by varying the parameters such as VCO concentration, enzyme loading, water content, and reaction temperature. It was found that VCO exhibited substrate inhibition at the concentration more than 40% (v/v). Lipozyme also achieved the highest production of free fatty acids, 4.56 mM at 1% (w/v) of enzyme loading. The optimum water content for VCO hydrolysis was 7% (v/v). A relatively high content of water was required because water was one of the reactants in the hydrolysis. The progress curve and the temperature profile of the enzymatic hydrolysis also showed that Lipozyme could be used for free fatty acid production at the temperature up to 50°C. However, the highest initial reaction rate and the highest yield of free fatty acid production were at 45 and 40°C, respectively. A 100 hours of initial reaction time has to be compensated in order to obtain the highest yield of free fatty acid production at 40°C.

Hydrolysis of Virgin Coconut Oil Using Immobilized Lipase in a Batch Reactor

PubMed Central

Chua, Lee Suan; Alitabarimansor, Meisam; Lee, Chew Tin; Mat, Ramli

2012-01-01

Hydrolysis of virgin coconut oil (VCO) had been carried out by using an immobilised lipase from Mucor miehei (Lipozyme) in a water-jacketed batch reactor. The kinetic of the hydrolysis was investigated by varying the parameters such as VCO concentration, enzyme loading, water content, and reaction temperature. It was found that VCO exhibited substrate inhibition at the concentration more than 40% (v/v). Lipozyme also achieved the highest production of free fatty acids, 4.56 mM at 1% (w/v) of enzyme loading. The optimum water content for VCO hydrolysis was 7% (v/v). A relatively high content of water was required because water was one of the reactants in the hydrolysis. The progress curve and the temperature profile of the enzymatic hydrolysis also showed that Lipozyme could be used for free fatty acid production at the temperature up to 50°C. However, the highest initial reaction rate and the highest yield of free fatty acid production were at 45 and 40°C, respectively. A 100 hours of initial reaction time has to be compensated in order to obtain the highest yield of free fatty acid production at 40°C. PMID:22953055

Pioneering fungi from the Damma glacier forefield in the Swiss Alps can promote granite weathering.

PubMed

Brunner, I; Plötze, M; Rieder, S; Zumsteg, A; Furrer, G; Frey, B

2011-05-01

Fungi were isolated from fine granitic sediments, which were collected at 15 sampling points within a 20 m × 40 m area in front of the Damma glacier in the central Swiss Alps. From the 45 fungal isolates grown on nutrient-rich agar media at 4 °C, 24 isolates were selected for partial sequencing and identification based on the small subunit ribosomal DNA. Sequencing data revealed that the isolated fungi represented three fungal phyla and 15 species. The weathering potential of 10 of the 15 fungal species was tested with dissolution experiments using powdered granite material (<63 μm). The results showed that the zygomyceteous species Mucor hiemalis, Umbelopsis isabellina and Mortierella alpina dissolved the granite powder most efficiently due to the release of a variety of organic acids, mainly citrate, malate and oxalate. In particular, the high concentrations of Ca, Fe, Mg and Mn in the solutions clustered well with the high amounts of exuded citrate. This is the first report on fungi that were isolated from a non-vegetated glacier forefield in which the fungi's capabilities to dissolve granite minerals were examined. © 2011 Blackwell Publishing Ltd.

Generation of efficient mutants of endoglycosidase from Streptococcus pyogenes and their application in a novel one-pot transglycosylation reaction for antibody modification

PubMed Central

Sekiguchi, Yukiko; Nakamura, Kensuke; Kawaguchi, Yoshirou; Honda, Takeshi; Hasegawa, Jun

2018-01-01

The fine structures of Fc N-glycan modulate the biological functions and physicochemical properties of antibodies. By remodeling N-glycan to obtain a homogeneous glycoform or chemically modified glycan, antibody characteristics can be controlled or modified. Such remodeling can be achieved by transglycosylation reactions using a mutant of endoglycosidase from Streptococcus pyogenes (Endo-S) and glycan oxazoline. In this study, we generated improved mutants of Endo-S by introducing additional mutations to the D233Q mutant. Notably, Endo-S D233Q/Q303L, D233Q/E350Q, and several other mutations resulted in transglycosylation efficiencies exceeding 90%, with a single-digit donor-to-substrate ratio of five, and D233Q/Y402F/D405A and several other mutations resulted in slightly reduced transglycosylation efficiencies accompanied by no detectable hydrolysis activity for 48 h. We further demonstrated that the combined use of mutants of Endo-S with Endo-M or Endo-CC, endoglycosidases from Mucor hiemalis and Coprinopsis cinerea, enables one-pot transglycosylation from sialoglycopeptide to antibodies. This novel reaction enables glycosylation remodeling of antibodies, without the chemical synthesis of oxazoline in advance or in situ. PMID:29474426

Generation of efficient mutants of endoglycosidase from Streptococcus pyogenes and their application in a novel one-pot transglycosylation reaction for antibody modification.

PubMed

Iwamoto, Mitsuhiro; Sekiguchi, Yukiko; Nakamura, Kensuke; Kawaguchi, Yoshirou; Honda, Takeshi; Hasegawa, Jun

2018-01-01

The fine structures of Fc N-glycan modulate the biological functions and physicochemical properties of antibodies. By remodeling N-glycan to obtain a homogeneous glycoform or chemically modified glycan, antibody characteristics can be controlled or modified. Such remodeling can be achieved by transglycosylation reactions using a mutant of endoglycosidase from Streptococcus pyogenes (Endo-S) and glycan oxazoline. In this study, we generated improved mutants of Endo-S by introducing additional mutations to the D233Q mutant. Notably, Endo-S D233Q/Q303L, D233Q/E350Q, and several other mutations resulted in transglycosylation efficiencies exceeding 90%, with a single-digit donor-to-substrate ratio of five, and D233Q/Y402F/D405A and several other mutations resulted in slightly reduced transglycosylation efficiencies accompanied by no detectable hydrolysis activity for 48 h. We further demonstrated that the combined use of mutants of Endo-S with Endo-M or Endo-CC, endoglycosidases from Mucor hiemalis and Coprinopsis cinerea, enables one-pot transglycosylation from sialoglycopeptide to antibodies. This novel reaction enables glycosylation remodeling of antibodies, without the chemical synthesis of oxazoline in advance or in situ.

Hygienic quality of traditional processing and stability of tomato (Lycopersicon esculentum) puree in Togo.

PubMed

Ameyapoh, Yaovi; de Souza, Comlan; Traore, Alfred S

2008-09-01

Microbiological and physicochemical qualities of a tomato (Lycopersicon esculentum) puree production line (ripe tomato, washing, cutting, pounding, bleaching, straining, bottling and pasteurization) and its preservation in Togo, West Africa, were studied using the HACCP method. Samples generated during the steps described previously were analyzed by determining sensory, chemical and microbiological characteristics. Samples were analyzed using MPN for coliform populations and plate count methodology for other bacteria. The microorganisms involved in spoilage of the opened products were moulds of genera Penicillium, Aspergillus, Fusarium, Geotrichum, Mucor and gram-positive Bacillus bacteria. The preserved tomato puree exhibited a pH value of 4.3, 90% water content, 0.98 water activity (aw) and an average ascorbic acid level of 27.3mg/100g. Results showed that the critical control point (CCP) of this tomato puree processing line is the pasteurization stage. The analysis of selected microbiological and physicochemical parameters during the preservation of bottled tomato puree indicated that this product was stable over 22 months at 29 degrees C. But the stability of the opened product stored at 29 degrees C did not exceed two months.

Genetic identification of the main opportunistic Mucorales by PCR-restriction fragment length polymorphism.

PubMed

Machouart, M; Larché, J; Burton, K; Collomb, J; Maurer, P; Cintrat, A; Biava, M F; Greciano, S; Kuijpers, A F A; Contet-Audonneau, N; de Hoog, G S; Gérard, A; Fortier, B

2006-03-01

Mucormycosis is a rare and opportunistic infection caused by fungi belonging to the order Mucorales. Recent reports have demonstrated an increasing incidence of mucormycosis, which is frequently lethal, especially in patients suffering from severe underlying conditions such as immunodeficiency. In addition, even though conventional mycology and histopathology assays allow for the identification of Mucorales, they often fail in offering a species-specific diagnosis. Due to the lack of other laboratory tests, a precise identification of these molds is thus notoriously difficult. In this study we aimed to develop a molecular biology tool to identify the main Mucorales involved in human pathology. A PCR strategy selectively amplifies genomic DNA from molds belonging to the genera Absidia, Mucor, Rhizopus, and Rhizomucor, excluding human DNA and DNA from other filamentous fungi and yeasts. A subsequent digestion step identified the Mucorales at genus and species level. This technique was validated using both fungal cultures and retrospective analyses of clinical samples. By enabling a rapid and precise identification of Mucorales strains in infected patients, this PCR-restriction fragment length polymorphism-based method should help clinicians to decide on the appropriate treatment, consequently decreasing the mortality of mucormycosis.

Genetic Identification of the Main Opportunistic Mucorales by PCR-Restriction Fragment Length Polymorphism

PubMed Central

Machouart, M.; Larché, J.; Burton, K.; Collomb, J.; Maurer, P.; Cintrat, A.; Biava, M. F.; Greciano, S.; Kuijpers, A. F. A.; Contet-Audonneau, N.; de Hoog, G. S.; Gérard, A.; Fortier, B.

2006-01-01

Mucormycosis is a rare and opportunistic infection caused by fungi belonging to the order Mucorales. Recent reports have demonstrated an increasing incidence of mucormycosis, which is frequently lethal, especially in patients suffering from severe underlying conditions such as immunodeficiency. In addition, even though conventional mycology and histopathology assays allow for the identification of Mucorales, they often fail in offering a species-specific diagnosis. Due to the lack of other laboratory tests, a precise identification of these molds is thus notoriously difficult. In this study we aimed to develop a molecular biology tool to identify the main Mucorales involved in human pathology. A PCR strategy selectively amplifies genomic DNA from molds belonging to the genera Absidia, Mucor, Rhizopus, and Rhizomucor, excluding human DNA and DNA from other filamentous fungi and yeasts. A subsequent digestion step identified the Mucorales at genus and species level. This technique was validated using both fungal cultures and retrospective analyses of clinical samples. By enabling a rapid and precise identification of Mucorales strains in infected patients, this PCR-restriction fragment length polymorphism-based method should help clinicians to decide on the appropriate treatment, consequently decreasing the mortality of mucormycosis. PMID:16517858

Chemodiversity and biodiversity of fungi associated with the pine weevil Hylobius abietis.

PubMed

Azeem, Muhammad; Terenius, Olle; Rajarao, Gunaratna Kuttuva; Nagahama, Kazuhiro; Nordenhem, Henrik; Nordlander, Göran; Borg-Karlson, Anna-Karin

2015-08-01

The pine weevil Hylobius abietis is a severe pest of conifer seedlings in reforestation areas. Weevils lay eggs in the root bark or in the soil near roots of recently dead trees and cover the eggs with frass (feces combined with chewed bark), possibly to avoid conspecific egg predation. The aim of the present investigation focused on isolation, identification, and volatile production of fungi from pine-weevil feces and frass. Fungi were isolated from weevil frass and feces separately, followed by identification based on ITS sequencing. Fifty-nine isolates belonging to the genera Penicillium, Ophiostoma, Mucor, Leptographium, Eucasphaeria, Rhizosphaera, Debaryomyces, and Candida were identified. Volatile organic compounds (VOCs) produced by the fungal community and fungal isolates cultured on weevil-frass broth were identified by SPME-GCMS. Major VOCs emitted from the fungal community and pure isolates were species- and strain specific and included isopentylalcohol, styrene, 3-octanone, 6-protoilludene, methyl salicylate, 3-methylanisole, 2-methoxyphenol, and phenol. Some of these are known to influence the orientation of pine weevils when tested among highly attractive newly planted conifer seedlings. Copyright © 2015 The British Mycological Society. Published by Elsevier Ltd. All rights reserved.

Xerophilic aflatoxigenic black tea fungi and their inhibition by Elettaria cardamomum and Syzygium aromaticum extracts

PubMed Central

Al-Sohaibani, Saleh; Murugan, K.; Lakshimi, G.; Anandraj, K.

2011-01-01

Black tea is consumed worldwide and is believed to play a role in cancer prevention. Xerophilic aflatoxigenic fungi are highly hazardous contaminants of tea since they are associated with tea quality impairment and human health risk. The present study reports isolation of such xerophilic and aflatoxigenic fungi associated with marketed tea. Twenty different tea samples collected from the local markets of Tamilnadu, India were investigated for fungal contamination. The results indicated contamination by 0.38% Aspergillus flavus. Other common contaminant fungi including Penicillium spp. (0.30%), Pacelomyces spp. (0.14%), and Mucor spp. (0.19%) were also isolated. Amongst the fungi isolated Aspergillus niger ML01 and A. flavus ML02 were found to be xerophilic aflatoxigenic mycoflora. Phylogenetic analysis based on 28S rRNA revealed their close ancestry. The chloroform and acetone extracts of spices Elettaria cardamomum and Syzygium aromaticum exhibited antifungal inhibitory activity on growth and toxin elaboration of both these xerophilic tea contaminants A. niger ML01 and A. flavus ML02. The results advocate the use of these spices plant or their extracts as novel antimicrobials which may add preservation and flavour in marketed tea. PMID:23961151

Fungicide contamination reduces beneficial fungi in bee bread based on an area-wide field study in honey bee, Apis mellifera, colonies.

PubMed

Yoder, Jay A; Jajack, Andrew J; Rosselot, Andrew E; Smith, Terrance J; Yerke, Mary Clare; Sammataro, Diana

2013-01-01

Fermentation by fungi converts stored pollen into bee bread that is fed to honey bee larvae, Apis mellifera, so the diversity of fungi in bee bread may be related to its food value. To explore the relationship between fungicide exposure and bee bread fungi, samples of bee bread collected from bee colonies pollinating orchards from 7 locations over 2 years were analyzed for fungicide residues and fungus composition. There were detectable levels of fungicides from regions that were sprayed before bloom. An organic orchard had the highest quantity and variety of fungicides, likely due to the presence of treated orchards within bees' flight range. Aspergillus, Penicillium, Rhizopus, and Cladosporium (beneficial fungi) were the primary fungal isolates found, regardless of habitat differences. There was some variation in fungal components amongst colonies, even within the same apiary. The variable components were Absidia, Alternaria, Aureobasidium, Bipolaris, Fusarium, Geotrichum, Mucor, Nigrospora, Paecilomyces, Scopulariopsis, and Trichoderma. The number of fungal isolates was reduced as an effect of fungicide contamination. Aspergillus abundance was particularly affected by increased fungicide levels, as indicated by Simpson's diversity index. Bee bread showing fungicide contamination originated from colonies, many of which showed chalkbrood symptoms.

Two classes of small antisense RNAs in fungal RNA silencing triggered by non-integrative transgenes

PubMed Central

Nicolás, Francisco E.; Torres-Martínez, Santiago; Ruiz-Vázquez, Rosa M.

2003-01-01

Transformation of Mucor circinelloides with self-replicative plasmids containing a wild-type copy of the carotenogenic gene carB causes silencing of the carB function in 3% of transformants. Genomic analyses revealed a relationship between silenced phenotype and number of copies of plasmids. This phenotype results from a reduction of the steady-state levels of carB mRNA, a reduction that is not due to differences in the level of transcription, indicating that silencing is post-transcriptional. Small sense and antisense RNAs have been found to be associated with gene silencing in M.circinelloides. Two size classes of small antisense RNAs, differentially accumulated during the vegetative growth of silenced transformants, have been detected: a long 25-nucleotide RNA and a short 21-nucleotide RNA. Secondary sense and antisense RNAs corresponding to sequences of the endogenous gene downstream of the initial triggering molecule have also been detected, revealing the existence of spreading of RNA targeting in fungi. These findings, together with the self-replicative nature of the triggering molecules, make M.circinelloides a suitable organism for investigating some unresolved questions in RNA silencing. PMID:12881432

Rich nutrition from the poorest - cereal fermentations in Africa and Asia.

PubMed

Nout, M J Rob

2009-10-01

Cereal fermentations in Africa and Asia involve mainly the processing of maize, rice, sorghum and the millets. Lactic acid bacteria (Lactobacillus, Pediococcus), Enterobacter spp., yeasts (Candida, Debaryomyces, Endomycopsis, Hansenula, Pichia, Saccharomyces and Trichosporon spp.) and filamentous fungi (Amylomyces, Aspergillus, Mucor, and Rhizopus spp.) contribute to desirable modifications of taste, flavour, acidity, digestibility, and texture in non-alcoholic beverages (e.g., uji, and ben-saalga), porridges (e.g., mawè) and cooked gels (e.g., kenkey, idli, and mifen). In addition, alcoholic beverages (beers such as tchoukoutou and jnard; and spirits e.g. jiu) are obtained using malt, or using amylolytic mixed microbial starter cultures as generators of fermentable substrates. Wet processing, marketing of multi-purpose intermediate products, co-fermentation for texture and nutrition, and mixed culture fermentations as practiced in indigenous fermentation processes are of interest for industrial innovation and for better control of natural mixed culture fermentation systems. On the other hand, the nutritional properties of traditional cereal fermented products can be enhanced by increasing their nutrient and energy density, as well as by increasing their mineral status by combining mineral fortification and dephytinization.

Assessment of aflatoxigenic Aspergillus and other fungi in millet and sesame from Plateau State, Nigeria

PubMed Central

Ezekiel, C.N.; Udom, I.E.; Frisvad, J.C.; Adetunji, M.C.; Houbraken, J.; Fapohunda, S.O.; Samson, R.A.; Atanda, O.O.; Agi-Otto, M.C.; Onashile, O.A.

2014-01-01

Sixteen fonio millet and 17 sesame samples were analysed for incidence of moulds, especially aflatoxigenic Aspergillus species, in order to determine the safety of both crops to consumers, and to correlate aflatoxin levels in the crops with levels produced by toxigenic isolates on laboratory medium. Diverse moulds including Alternaria, Aspergillus, Cercospora, Fusarium, Mucor, Penicillium, Rhizopus and Trichoderma were isolated. Aspergillus was predominantly present in both crops (46–48%), and amongst the potentially aflatoxigenic Aspergillus species, A. flavus recorded the highest incidence (68% in fonio millet; 86% in sesame kernels). All A. parvisclerotigenus isolates produced B and G aflatoxins in culture while B aflatoxins were produced by only 39% and 20% of A. flavus strains isolated from the fonio millet and sesame kernels, respectively. Aflatoxin concentrations in fonio millet correlated inversely (r = −0.55; p = 0.02) with aflatoxin levels produced by toxigenic isolates on laboratory medium, but no correlation was observed in the case of the sesame samples. Both crops, especially sesame, may not be suitable substrates for aflatoxin biosynthesis. This is the first report on A. parvisclerotigenus in sesame. PMID:24772370

Anion effects on anti-microbial activity of poly[1-vinyl-3-(2-sulfoethyl imidazolium betaine)].

PubMed

Garg, Godawari; Chauhan, Ghanshyam S; Gupta, Reena; Ahn, J-H

2010-04-01

Recent investigations in the anti-microbial properties of the functional polymers are predominantly focused on the structure of the cationic moieties. In the present study, we investigated that the nature of the anion present in polysulfobetaines affects activity against certain microorganisms and their anti-microbial properties have been rationalized in terms of the structure-activity relationship. Vinyl imidazolium-based polysulfobetaines were prepared by the quaternization of poly(N-vinyl imidazole) with sodium salt of 2-bromo ethanesulfonic acid. The bromide counter anion of the resulting polymer was exchanged with different anions to generate a series of polymers. These were characterized by FTIR, DSC, XRD, SEM, elemental analysis (C, H, N and S) and viscosity measurements. The anti-microbial activity studies were carried against three fungi (Aspergillus niger, Byssochlamys fulva and Mucor circenelliods) and two bacteria (Bacillus coagulans BTS-3 and Pseudomonas aeruginosa BTS-2). The nature of the anion affects the structure of polysulfobetaine by realignment of polymer chains. The anion-dependent anti-microbial properties of polysulfobetaines result from the interaction of the microbes at the polymer interface. Copyright 2009 Elsevier Inc. All rights reserved.

Bioprospecting of lipolytic microorganisms obtained from industrial effluents.

PubMed

Peil, Greice H S; Kuss, Anelise V; Rave, Andrés F G; Villarreal, José P V; Hernandes, Yohana M L; Nascente, Patrícia S

2016-01-01

The lipases have ability to catalyze diverse reactions and are important in different biotechnological applications. The aim of this work was to isolate and characterize microorganisms that produce lipases, from different food industry effluents localized in Pelotas, RS/Brazil. Bacteria were identified using Gram stain and biochemical tests (Vitek 2(r)). Fungi were identified according to macro and micromorphology characteristics. The extracellular lipase production was evaluated using the Rhodamine B test and the enzymatic activity by titration. Twenty-one bacteria were isolated and identified as Klebsiella pneumoniae ssp. pneumoniae, Serratia marcescens, Enterobacter aerogenes, Raoultella ornithinolytica and Raoultella planticola. Were characterized isolated filamentous fungi by the following genera: Alternaria sp., Fusarium sp., Geotrichum sp., Gliocladium sp., Mucor sp., Paecilomyces sp. and Trichoderma sp. Extracellular lipase production was observed in 71.43% of the bacteria and 57.14% of the fungi. The bacterium that presented better promising enzymatic activity was E. aerogenes (1.54 U/ml) however between fungi there was not significant difference between the four isolates. This study indicated that microorganisms lipase producers are present in the industrial effluents, as well as these enzymes have potential of biodegradation of lipid compounds.

Colony-PCR Is a Rapid Method for DNA Amplification of Hyphomycetes

PubMed Central

Walch, Georg; Knapp, Maria; Rainer, Georg; Peintner, Ursula

2016-01-01

Fungal pure cultures identified with both classical morphological methods and through barcoding sequences are a basic requirement for reliable reference sequences in public databases. Improved techniques for an accelerated DNA barcode reference library construction will result in considerably improved sequence databases covering a wider taxonomic range. Fast, cheap, and reliable methods for obtaining DNA sequences from fungal isolates are, therefore, a valuable tool for the scientific community. Direct colony PCR was already successfully established for yeasts, but has not been evaluated for a wide range of anamorphic soil fungi up to now, and a direct amplification protocol for hyphomycetes without tissue pre-treatment has not been published so far. Here, we present a colony PCR technique directly from fungal hyphae without previous DNA extraction or other prior manipulation. Seven hundred eighty-eight fungal strains from 48 genera were tested with a success rate of 86%. PCR success varied considerably: DNA of fungi belonging to the genera Cladosporium, Geomyces, Fusarium, and Mortierella could be amplified with high success. DNA of soil-borne yeasts was always successfully amplified. Absidia, Mucor, Trichoderma, and Penicillium isolates had noticeably lower PCR success. PMID:29376929

Fungal flora associated with combine harvester wheat and sorghum dusts from Egypt.

PubMed

Abdel-Hafez, S I; Moubasher, A H; Shoreit, A A; Ismail, M A

1990-01-01

107 species and 8 species varieties belonging to 44 genera were collected from combine harvester wheat and sorghum dusts (35 genera and 91 species + 4 varieties) and from the atmosphere of their hay sites (26 genera and 69 species + 4 varieties) on glucose- and cellulose-Czapek's Dox agar at 28 degrees C and 45 degrees C. The mycoflora of wheat and sorghum dusts were basically similar on the two types of media and the most common fungi were: Alternaria alternata, Aspergillus flavus, A. fumigatus, A. niger, A. ochraceus, A. sydowii, A. terreus, Cochliobolus spicifer, Emericella nidulans, Fusarium moniliforme, Penicillium chrysogenum, P. duclauxii, P. funiculosum and P. oxalicum. Truly thermophilic species were frequently encountered from the two substrates: Chaetomium thermophilum, Humicola grisea var. thermoidae, H. insolens, Malbranchea pulchella var., sulphurea, Rhizomucor pusillus, Sporotrichum thermophilum, Talaromyces thermophilus, Thermoascus thermophilus and Thermomyces lanuginosus. The airborne fungi in the two atmospheres were basically similar and the most prevalent species were members of Alternaria (1 species), Aspergillus (18 species + 2 varieties), Chaetomium (2 species), Cochliobolus(3 species), Emericella (3 species + 2 varieties), Fusarium (3 species), Mucor (1 species), Penicillium (14 species) and Stachybotrys (1 species).

Association between sensitization to Aureobasidium pullulans (Pullularia sp) and severity of asthma.

PubMed

Niedoszytko, Marek; Chełmińska, Marta; Jassem, Ewa; Czestochowska, Eugenia

2007-02-01

Recent data indicate that fungi may contribute to increased severity of asthma. To determine the prevalence of allergy to 15 mold allergens among patients hospitalized because of exacerbation of asthma and to evaluate the relationship between the severity of the disease and allergy to particular molds. Skin prick tests with standard aeroallergens of airborne allergens, including grass, tree, Dermatophagoides pteronyssinus, Dermatophagoides farinae, feather, and cat and dog fur, and a panel of mold allergens, including Alternaria, Cladosporium, Aspergillus, Penicillium, Trichothecium, Chaetomium globosum, Epicoccum, Epidermophyton, Helminthosporium, Aureobasidium pullulans, Rhizopus nigricans, Fusarium, Mucor, Merulius lacrymans, and yeast mix, were performed in 105 asthmatic patients and 30 controls. Positive skin prick test results were found in 98% of asthmatic patients and 66% of controls. Sensitivity to A pullulans was significantly associated with more severe asthma (odds ratio, 1.4; 95% confidence interval, 1.09-1.75; P = .006). Sensitization to Helminthosporium was associated with an increased number of asthma exacerbations that required hospitalization (17% vs 38%; chi2 test P = .03). Sensitization to A pullulans is a risk factor for severe asthma. Sensitization to Helminthosporium may be related to asthma exacerbation that requires hospitalization.

DNA-binding studies and biological activities of new nitrosubstituted acyl thioureas

NASA Astrophysics Data System (ADS)

Tahir, Shaista; Badshah, Amin; Hussain, Raja Azadar; Tahir, Muhammad Nawaz; Tabassum, Saira; Patujo, Jahangir Ali; Rauf, Muhammad Khawar

2015-11-01

Four new nitrosubstituted acylthioureas i.e. 1-acetyl-3-(4-nitrophenyl)thiourea (TU1), 1-acetyl-3-(2-methyl-4-nitrophenyl)thiourea (TU2), 1-acetyl-3-(2-methoxy-4-nitrophenyl)thiourea (TU3) and 1-acetyl-3-(4-chloro-3-nitrophenyl)thiourea (TU4) have been synthesized and characterized (by C13 and H1 nuclear magnetic resonance, Fourier transform infrared spectroscopy and single crystal X-ray diffraction). As a preliminary investigation of the anti-cancer potencies of the said compounds, DNA interaction studies have been carried out using cyclic voltammetry and UV-vis spectroscopy along with verification from computational studies. The drug-DNA binding constants are found to be in the order, KTU3 9.04 × 106 M-1 > KTU4 8.57 × 106 M-1 > KTU2 6.05 × 106 M-1 > KTU1 1.16 × 106 M-1. Furthermore, the antioxidant, cytotoxic, antibacterial and antifungal activities have been carried out against DPPH (1,1-diphenyl-2-dipicrylhydrazyl), Brine shrimp eggs, gram positive (Micrococcus luteus, Staphylococcus aureus) and gram negative (Bordetella bronchiseptica, Salmonella typhimurium, Enterobacter aerogens) and fungal cultures (Aspergillus fumigatus, Mucor species, Aspergillus niger, Aspergillus flavus) respectively.

Scanning electron and light microscopic study of microbial succession on bethlehem st. Nectaire cheese.

PubMed

Marcellino, S N; Benson, D R

1992-11-01

St. Nectaire cheese is a semisoft cheese of French origin that, along with Brie and Camembert cheeses, belongs to the class of surface mold-ripened cheese. The surface microorganisms that develop on the cheese rind during ripening impart a distinctive aroma and flavor to this class of cheese. We have documented the sequential appearance of microorganisms on the cheese rind and in the curd over a 60-day ripening period. Scanning electron microscopy was used to visualize the development of surface fungi and bacteria. Light microscopy of stained paraffin sections was used to study cross sections through the rind. We also monitored the development of bacterial and yeast populations in and the pH of the curd and rind. The earliest stage of ripening (0 to 2 days) is dominated by the lactic acid bacterium Streptococcus cremoris and multilateral budding yeasts, primarily Debaryomyces and Torulopsis species. Geotrichum candidum follows closely, and then zygomycetes of the genus Mucor develop at day 4 of ripening. At day 20, the deuteromycete Trichothecium roseum appears. From day 20 until the end of the ripening process, coryneforms of the genera Brevibacterium and Arthrobacter can be seen near the surface of the cheese rind among fungal hyphae and yeast cells.

Antifungal Activity of Lactobacillus sp. Bacteria in the Presence of Xylitol and Galactosyl-Xylitol

PubMed Central

Lipińska, Lidia; Klewicki, Robert; Klewicka, Elżbieta; Kołodziejczyk, Krzysztof; Sójka, Michał; Nowak, Adriana

2016-01-01

Lactic acid fermentation is a natural method of antimicrobial food protection. Antagonistic activity of Lactobacillus sp. bacteria, taking part in this process, is directed mainly against the same or other microorganisms. In this work we determine the impact of the presence of xylitol and galactosyl-xylitol on the antagonistic activity of 60 Lactobacillus sp. strains against indicator molds (Alternaria alternata, Alternaria brassicicola, Aspergillus niger, Fusarium latenicum, Geotrichum candidum, and Mucor hiemalis) and yeasts (Candida vini). We used double-layer method to select antifungal strains of Lactobacillus bacteria and poisoned medium method to confirm their fungistatic properties. Additionally, we examined the inhibition of Alternaria brassicicola by Lactobacillus paracasei ŁOCK 0921 cultivated with xylitol or galactosyl-xylitol directly on wild cherries. The presence of xylitol and its galactosyl derivative led to increase of spectrum of antifungal activity in most of the studied plant-associated lactobacilli strains. However, no single strain exhibited activity against all the indicator microorganisms. The antifungal activity of Lactobacillus bacteria against molds varied considerably and depended on both the indicator strain and the composition of the medium. The presence of xylitol and galactosyl-xylitol in the growth medium is correlated with the antifungal activity of the studied Lactobacillus sp. bacteria against selected indicator molds. PMID:27294124

Proteomic Analysis of Pathogenic Fungi Reveals Highly Expressed Conserved Cell Wall Proteins

PubMed Central

Champer, Jackson; Ito, James I.; Clemons, Karl V.; Stevens, David A.; Kalkum, Markus

2016-01-01

We are presenting a quantitative proteomics tally of the most commonly expressed conserved fungal proteins of the cytosol, the cell wall, and the secretome. It was our goal to identify fungi-typical proteins that do not share significant homology with human proteins. Such fungal proteins are of interest to the development of vaccines or drug targets. Protein samples were derived from 13 fungal species, cultured in rich or in minimal media; these included clinical isolates of Aspergillus, Candida, Mucor, Cryptococcus, and Coccidioides species. Proteomes were analyzed by quantitative MSE (Mass Spectrometry—Elevated Collision Energy). Several thousand proteins were identified and quantified in total across all fractions and culture conditions. The 42 most abundant proteins identified in fungal cell walls or supernatants shared no to very little homology with human proteins. In contrast, all but five of the 50 most abundant cytosolic proteins had human homologs with sequence identity averaging 59%. Proteomic comparisons of the secreted or surface localized fungal proteins highlighted conserved homologs of the Aspergillus fumigatus proteins 1,3-β-glucanosyltransferases (Bgt1, Gel1-4), Crf1, Ecm33, EglC, and others. The fact that Crf1 and Gel1 were previously shown to be promising vaccine candidates, underlines the value of the proteomics data presented here. PMID:26878023

The Celecoxib Derivative AR-12 Has Broad-Spectrum Antifungal Activity In Vitro and Improves the Activity of Fluconazole in a Murine Model of Cryptococcosis

PubMed Central

Koselny, Kristy; Green, Julianne; DiDone, Louis; Halterman, Justin P.; Fothergill, Annette W.; Wiederhold, Nathan P.; Patterson, Thomas F.; Cushion, Melanie T.; Rappelye, Chad; Wellington, Melanie

2016-01-01

Only one new class of antifungal drugs has been introduced into clinical practice in the last 30 years, and thus the identification of small molecules with novel mechanisms of action is an important goal of current anti-infective research. Here, we describe the characterization of the spectrum of in vitro activity and in vivo activity of AR-12, a celecoxib derivative which has been tested in a phase I clinical trial as an anticancer agent. AR-12 inhibits fungal acetyl coenzyme A (acetyl-CoA) synthetase in vitro and is fungicidal at concentrations similar to those achieved in human plasma. AR-12 has a broad spectrum of activity, including activity against yeasts (e.g., Candida albicans, non-albicans Candida spp., Cryptococcus neoformans), molds (e.g., Fusarium, Mucor), and dimorphic fungi (Blastomyces, Histoplasma, and Coccidioides) with MICs of 2 to 4 μg/ml. AR-12 is also active against azole- and echinocandin-resistant Candida isolates, and subinhibitory AR-12 concentrations increase the susceptibility of fluconazole- and echinocandin-resistant Candida isolates. Finally, AR-12 also increases the activity of fluconazole in a murine model of cryptococcosis. Taken together, these data indicate that AR-12 represents a promising class of small molecules with broad-spectrum antifungal activity. PMID:27645246

Antifungal Activity of Lactobacillus sp. Bacteria in the Presence of Xylitol and Galactosyl-Xylitol.

PubMed

Lipińska, Lidia; Klewicki, Robert; Klewicka, Elżbieta; Kołodziejczyk, Krzysztof; Sójka, Michał; Nowak, Adriana

2016-01-01

Lactic acid fermentation is a natural method of antimicrobial food protection. Antagonistic activity of Lactobacillus sp. bacteria, taking part in this process, is directed mainly against the same or other microorganisms. In this work we determine the impact of the presence of xylitol and galactosyl-xylitol on the antagonistic activity of 60 Lactobacillus sp. strains against indicator molds (Alternaria alternata, Alternaria brassicicola, Aspergillus niger, Fusarium latenicum, Geotrichum candidum, and Mucor hiemalis) and yeasts (Candida vini). We used double-layer method to select antifungal strains of Lactobacillus bacteria and poisoned medium method to confirm their fungistatic properties. Additionally, we examined the inhibition of Alternaria brassicicola by Lactobacillus paracasei ŁOCK 0921 cultivated with xylitol or galactosyl-xylitol directly on wild cherries. The presence of xylitol and its galactosyl derivative led to increase of spectrum of antifungal activity in most of the studied plant-associated lactobacilli strains. However, no single strain exhibited activity against all the indicator microorganisms. The antifungal activity of Lactobacillus bacteria against molds varied considerably and depended on both the indicator strain and the composition of the medium. The presence of xylitol and galactosyl-xylitol in the growth medium is correlated with the antifungal activity of the studied Lactobacillus sp. bacteria against selected indicator molds.

Optimized synthesis of lipase-catalyzed hexyl acetate in n-hexane by response surface methodology.

PubMed

Shieh, C J; Chang, S W

2001-03-01

Hexyl acetate, a short-chain ester with fruity odor, is a significant green note flavor compound and widely used in the food industry. The ability for immobilized lipase from Mucor miehei (Lipozyme IM-77) to catalyze the transesterification of hexanol with triacetin was investigated in this study. Response surface methodology and five-level-five-factor central composite rotatable design were adopted to evaluate the effects of synthesis variables, such as reaction time (2-10 h), temperature (25-65 degrees C), enzyme amount (10-50%; 0.024-0.118 BAUN), substrate molar ratio of triacetin to hexanol (1:1 to 3:1), and added water content (0-20%) on percentage molar conversion of hexyl acetate. The results showed that reaction temperature and substrate molar ratio were the most important parameters and that added water content had less of an effect on percent molar conversion. On the basis of canonical analysis, optimum synthesis conditions were as follows: reaction time, 7.7 h; temperature, 52.6 degrees C; enzyme amount, 37.1% (0.089 BAUN); substrate molar ratio, 2.7:1; and added water, 12.5%. The predicted value was 88.9% molar conversion, and the actual experimental value was 86.6% molar conversion.

Estimating biodiversity of fungi in activated sludge communities using culture-independent methods.

PubMed

Evans, Tegan N; Seviour, Robert J

2012-05-01

Fungal diversity of communities in several activated sludge plants treating different influent wastes was determined by comparative sequence analyses of their 18S rRNA genes. Methods for DNA extraction and choice of primers for PCR amplification were both optimised using denaturing gradient gel electrophoresis profile patterns. Phylogenetic analysis revealed that the levels of fungal biodiversity in some communities, like those treating paper pulp wastes, were low, and most of the fungi detected in all communities examined were novel uncultured representatives of the major fungal subdivisions, in particular, the newly described clade Cryptomycota. The fungal populations in activated sludge revealed by these culture-independent methods were markedly different to those based on culture-dependent data. Members of the genera Penicillium, Cladosporium, Aspergillus and Mucor, which have been commonly identified in mixed liquor, were not identified in any of these plant communities. Non-fungal eukaryotic 18S rRNA genes were also amplified with the primer sets used. This is the first report where culture-independent methods have been applied to flocculated activated sludge biomass samples to estimate fungal community composition and, as expected, the data obtained gave a markedly different view of their population biodiversity compared to that based on culture-dependent methods.

Anthropogenic impact on environmental filamentous fungi communities along the Mediterranean littoral.

PubMed

Al-Yasiri, Mohammed Hashim; Normand, Anne-Cécile; Mauffrey, Jean-François; Ranque, Stéphane

2017-07-01

We hypothesised that anthropogenic influences impact the filamentous fungi community structure and that particular species or species patterns might serve as markers to characterise ecosystems. This study aimed to describe the filamentous fungi community structure in various biotopes along the Mediterranean shore that were exposed to various levels of anthropogenic influence. We sampled filamentous fungi from yellow-legged gull faecal samples at five study sites along the Mediterranean littoral in southern France. The sites were characterised by variable anthropogenic influence, ranging from building rooftops in two cities to a natural reserve. The sites also included two suburban ecoclines, one of which was exposed to sewer pollution. Filamentous fungal colonies were quantified and identified via MALDI-TOF mass spectrometry. Interestingly, we found that both fungal diversity and abundance were low in urban areas compared with suburban ecocline or environments little affected by anthropogenic influence. Furthermore, some fungal species were clearly associated with particular environments. In particular, Mucor circinelloides was associated with a natural environment with little anthropogenic impact and distant from human settlements. Whereas, Scedosporium apiospermum was associated with an ecocline polluted by sewage. Our findings indicate that particular fungal species or species combination might be used as surrogate markers of ecosystems exposed to anthropogenic pollution. © 2017 Blackwell Verlag GmbH.

Metagenomics analysis of microbial communities associated with a traditional rice wine starter culture (Xaj-pitha) of Assam, India.

PubMed

Bora, Sudipta Sankar; Keot, Jyotshna; Das, Saurav; Sarma, Kishore; Barooah, Madhumita

2016-12-01

This is the first report on the microbial diversity of xaj-pitha, a rice wine fermentation starter culture through a metagenomics approach involving Illumine-based whole genome shotgun (WGS) sequencing method. Metagenomic DNA was extracted from rice wine starter culture concocted by Ahom community of Assam and analyzed using a MiSeq ® System. A total of 2,78,231 contigs, with an average read length of 640.13 bp, were obtained. Data obtained from the use of several taxonomic profiling tools were compared with previously reported microbial diversity studies through the culture-dependent and culture-independent method. The microbial community revealed the existence of amylase producers, such as Rhizopus delemar, Mucor circinelloides, and Aspergillus sp. Ethanol producers viz., Meyerozyma guilliermondii, Wickerhamomyces ciferrii, Saccharomyces cerevisiae, Candida glabrata, Debaryomyces hansenii, Ogataea parapolymorpha, and Dekkera bruxellensis, were found associated with the starter culture along with a diverse range of opportunistic contaminants. The bacterial microflora was dominated by lactic acid bacteria (LAB). The most frequent occurring LAB was Lactobacillus plantarum, Lactobacillus brevis, Leuconostoc lactis, Weissella cibaria, Lactococcus lactis, Weissella para mesenteroides, Leuconostoc pseudomesenteroides, etc. Our study provided a comprehensive picture of microbial diversity associated with rice wine fermentation starter and indicated the superiority of metagenomic sequencing over previously used techniques.

Pretreatment of different food rest materials for bioconversion into fungal lipid-rich biomass.

PubMed

Tzimorotas, D; Afseth, N K; Lindberg, D; Kjørlaug, O; Axelsson, L; Shapaval, V

2018-04-13

Food rest materials have the potential to be used as media components in various types of fermentations. Oleaginous filamentous fungi can utilize those components and generate a high-value lipid-rich biomass, which could be further used for animal and human use. One of the main limitations in this process is the pretreatment of food rest materials, needed to provide homogenization, sterilization and solubilization. In this study, two pretreatment processes-steam explosion and enzymatic hydrolysis-were evaluated for potato and animal protein-rich food rest materials. The pretreated food rest materials were used for the production of fungal lipid-rich biomass in submerged fermentation by the oleaginous fungus Mucor circinelloides. Cultivation media based on malt extract broth and glucose were used as controls of growth and lipid production, respectively. It was observed that media based on food rest materials can support growth and lipid production in M. circinelloides to a similar extent as the control media. More specifically, the use of potato hydrolysate combined with chicken auto-hydrolysate resulted in a higher fungal total biomass weight than using malt extract broth. When the same C/N ratio was used for glucose and rest materials-based media, similar lipid content was obtained or even higher using the latter media.

Sterol Composition of Clinically Relevant Mucorales and Changes Resulting from Posaconazole Treatment.

PubMed

Müller, Christoph; Neugebauer, Thomas; Zill, Patrizia; Lass-Flörl, Cornelia; Bracher, Franz; Binder, Ulrike

2018-05-19

Mucorales are fungi with increasing importance in the clinics. Infections take a rapidly progressive course resulting in high mortality rates. The ergosterol biosynthesis pathway and sterol composition are of interest, since they are targeted by currently applied antifungal drugs. Nevertheless, Mucorales often exhibit resistance to these drugs, resulting in therapeutic failure. Here, sterol patterns of six clinically relevant Mucorales ( Lichtheimia corymbifera , Lichtheimia ramosa , Mucor circinelloides , Rhizomucor pusillus , Rhizopus arrhizus , and Rhizopus microsporus ) were analysed in a targeted metabolomics fashion after derivatization by gas chromatography-mass spectrometry. Additionally, the effect of posaconazole (POS) treatment on the sterol pattern of R. arrhizus was evaluated. Overall, fifteen different sterols were detected with species dependent variations in the total and relative sterol amount. Sterol analysis from R. arrhizus hyphae confronted with sublethal concentrations of posaconazole revealed the accumulation of 14-methylergosta-8,24-diene-3,6-diol, which is a toxic sterol that was previously only detected in yeasts. Sterol content and composition were further compared to the well-characterized pathogenic mold Aspergillus fumigatus . This work contributes to a better understanding of the ergosterol biosynthesis pathway of Mucorales, which is essential to improve antifungal efficacy, the identification of targets for novel drug design, and to investigate the combinatorial effects of drugs targeting this pathway.

Keratinophilic fungi and other moulds associated with air-dust particles from Egypt.

PubMed

Abdel-Hafez, S I; Moubasher, A H; Barakat, A

1990-01-01

One-hundred and eleven species and three species varieties belonging to 39 genera were collected from 50 dust samples on the five media used at 28 degrees C. Using the hair-baiting technique with horse hair, 10 species of Chrysosporium were isolated: C. asperatum, C. state of Arthroderma tuberculatum, C. indicum, C. inops, C. keratinophilum, C. merdarium, C. pannorum, C. queenslandicum, C. tropicum and C. xerophilum. True dermatophytes were isolated: Trichophyton verrucosum and Trichophyton sp. Also, numerous fungi tolerating high levels of cycloheximide were encountered, such as members of Acremonium, Aspergillus and Penicillium. On plates of glucose or cellulose Czapek-Dox agar (free from sucrose) the most frequent fungi were: Alternaria alternata, Aspergillus flavus, A. flavus var. columnaris, A. fumigatus, A. niger, A. ochraceus, A. sydowii, A. terreus, Chaetomium globosum, Cladosporium herbarum, Emericella nidulans, Fusarium oxysporum, Mucor hiemalis, Penicillium chrysogenum, P. oxalicum, Scopulariopsis brevicaulis and Ulocladium atrum. On plates of 50% sucrose or 10 and 20% NaCl-Czapek's agar, some interesting species were frequently encountered: Eurotium amstelodami, E. chevalieri, E. halophilicum, E. montevidensis, E. repens, E. rubrum and Scopulariopsis halophilica. The isolated fungi have been tested for osmophilicity and halophilicity, they showed different rates of growth on sucrose and sodium chloride-Czapek's medium of various osmotic potential.

Further studies on Egyptian soil fungi: succession of sugar and osmophilic fungi in soil amended with five organic substrates.

PubMed

Shaban, G M

1996-01-01

The sugar and osmophilic fungal composition of soils amended with five organic substrates (newspaper, orange peel, bromegrass leaves, wheat straw and wood sawdust) was estimated after 2, 4, 6, 8 and 10 weeks using the dilution plate method on glucose and 50% sucrose Czapek's agar media. Wheat straw was the best substrate for total counts of both sugar and osmophilic fungi followed by newspaper, bromegrass leaves, wood sawdust and orange peel. Wood sawdust supported the highest average counts of total sugar fungi, Fusarium, Mucor, Scopulariopsis, Trichoderma and Trimmatostroma spp.; Newspaper, of Aspergillus (8 spp.), Penicillium (4 spp.) and Chaetomium sp.; bromegrass leaves of Cladosporium sp., Humicola sp. and Sporotrichum sp.; orange peel, of Alternaria sp., Circinella sp. and Stachybotrys sp.; and wheat straw, of Botryotrichum sp. and Myrothecium sp. Bromegrass leaves and orange peel supported the highest average counts of total osmophilic fungi, Aspergillus (10 spp.), Cladosporium sp. Paecillomyces sp. and Rhizopus sp.; and of Stemphylium sp., Trichoderma sp., Humicola sp. and Circinella sp. respectively; wheat straw, of Epicoccum sp., Scopulariopsis sp. and Trichothecium sp.; newspaper, of Penicillium (4 spp.) and Alternaria sp.; and wood sawdust of Curvularia sp. and Fusarium (3 spp.). The best colonizers throughout the experimental periods were Aspergilus and Penicillium spp.

Microfungi in cultivated fields in Eskişehir provience (Turkey).

PubMed

Demirel, Rasime; Ilhan, Semra; Asan, Ahmet; Kinaci, Engin; Oner, Setenay

2005-01-01

The soil microfungi flora was investigated in four locations of Eskişehir (Turkey). 56 soil samples were seasonaly collected from 14 stations in the areas of Karacahöyük, Bahçecik, OGU I, and OGU II. A total of 110 species belonging to 32 genera were encountered including Absidia, Acremonium, Alternaria, Aspergillus, Beauveria, Botryoderma, Chaetomium, Chrysosporium, Cladosporium, Eupenicillium, Eurotium, Fusarium, Geotrichum, Gliocladium, Gonytrichum, Metarrhizium, Mucor, Myrothecium, Paecilomyces, Penicillium, Phoma, Plectosphaerella, Rhizoctania, Rhizopus, Scopulariopsis, Septonema, Stachybotrys, Trichocladium, Trichoderma, Ulocladium, Verticillium, and Wardomyces. Twenty five species were more frequent (all locations) while twenty seven species were rare (only one sample). Mainly, Acremonium kiliense, Aspergillus ochraceus, A. terricola var. americanus, A. versicolor, Cladosporium cladosporioides, Fusarium oxysporum, F. solani, Gliocladium roseum, Penicillium chrysogenum, P. corylophum, P. expansum, P. griseofulvum, P. implicatum, P. restrictum, and Stachybotrys chartarum were the most common and abundant microfungi in all locations. Five species Aspergillus subsessilis, A. terreus var. africanus, Eupenicillium egyptiacum, Paecilomyces ramosus, and Penicillium novae-zeelandiae are likely to be newly recorded for Turkey. The microfungi number in Eskişehir soils was between 25,000-234,000 CFU/g (mean value at 126,375 CFU/g). Copyright (c) 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Diversity of plant oil seed-associated fungi isolated from seven oil-bearing seeds and their potential for the production of lipolytic enzymes.

PubMed

Venkatesagowda, Balaji; Ponugupaty, Ebenezer; Barbosa, Aneli M; Dekker, Robert F H

2012-01-01

Commercial oil-yielding seeds (castor, coconut, neem, peanut, pongamia, rubber and sesame) were collected from different places in the state of Tamil Nadu (India) from which 1279 endophytic fungi were isolated. The oil-bearing seeds exhibited rich fungal diversity. High Shannon-Index H' was observed with pongamia seeds (2.847) while a low Index occurred for coconut kernel-associated mycoflora (1.018). Maximum Colonization Frequency (%) was observed for Lasiodiplodia theobromae (176). Dominance Index (expressed in terms of the Simpson's Index D) was high (0.581) for coconut kernel-associated fungi, and low for pongamia seed-borne fungi. Species Richness (Chao) of the fungal isolates was high (47.09) in the case of neem seeds, and low (16.6) for peanut seeds. All 1279 fungal isolates were screened for lipolytic activity employing a zymogram method using Tween-20 in agar. Forty isolates showed strong lipolytic activity, and were morphologically identified as belonging to 19 taxa (Alternaria, Aspergillus, Chalaropsis, Cladosporium, Colletotrichum, Curvularia, Drechslera, Fusarium, Lasiodiplodia, Mucor, Penicillium, Pestalotiopsis, Phoma, Phomopsis, Phyllosticta, Rhizopus, Sclerotinia, Stachybotrys and Trichoderma). These isolates also exhibited amylolytic, proteolytic and cellulolytic activities. Five fungal isolates (Aspergillus niger, Chalaropsis thielavioides, Colletotrichum gloeosporioides, Lasiodiplodia theobromae and Phoma glomerata) exhibited highest lipase activities, and the best producer was Lasiodiplodia theobromae (108 U/mL), which was characterized by genomic sequence analysis of the ITS region of 18S rDNA.

Volatile organic compounds from fungi isolated after hurricane katrina induce developmental defects and apoptosis in a Drosophila melanogaster model.

PubMed

Inamdar, Arati A; Bennett, Joan W

2015-05-01

In previous work, our laboratory developed a Drosophila model for studying the adverse effects of fungal volatile organic compounds (VOCs) emitted by growing cultures of molds. In this report, we have extended these studies and compared the toxic effects of fungal VOCs emitted from living cultures of four molds isolated after Hurricane Katrina from a flooded home in New Orleans. Strains of Aspergillus, Mucor, Penicillium, and Trichoderma were grown with wild-type larvae and the toxic effects of volatile products on the developmental stages of Drosophila larvae were evaluated. Furthermore, heterozygous mutants of Drosophila carrying the apoptotic genes, reaper and dronc, were used to assess the role of apoptosis in fungal VOCs mediated toxicity. Third-instar larvae of Drosophila carrying these apoptotic genes were exposed to fungal VOCs emitted from growing mold cultures for 10 days. The larval strains carrying apoptopic genes survived longer than the control wild type larvae; moreover, of those that survived, heterozygous reaper and dronc strains progressed to pupae and adult phases more rapidly, suggesting that fungal VOCs may induce apoptotic changes in flies. These data lend support to the use of Drosophila as an inexpensive and genetically versatile toxicological model to investigate the mechanistic basis for some of the human illnesses/symptoms associated with exposure to mold-contaminated indoor air, especially after hurricanes. © 2013 Wiley Periodicals, Inc.

Microbiological destruction of composite polymeric materials in soils

NASA Astrophysics Data System (ADS)

Legonkova, O. A.; Selitskaya, O. V.

2009-01-01

Representatives of the same species of microscopic fungi developed on composite materials with similar polymeric matrices independently from the type of soils, in which the incubation was performed. Trichoderma harzianum, Penicillium auranthiogriseum, and Clonostachys solani were isolated from the samples of polyurethane. Fusarium solani, Clonostachys rosea, and Trichoderma harzianum predominated on the surface of ultrathene samples. Ulocladium botrytis, Penicillium auranthiogriseum, and Fusarium solani predominated in the variants with polyamide. Trichoderma harzianum, Penicillium chrysogenum, Aspergillus ochraceus, and Acremonium strictum were isolated from Lentex-based composite materials. Mucor circinelloides, Trichoderma harzianum, and Penicillium auranthiogriseum were isolated from composite materials based on polyvinyl alcohol. Electron microscopy demonstrated changes in the structure of polymer surface (loosening and an increase in porosity) under the impact of fungi. The physicochemical properties of polymers, including their strength, also changed. The following substances were identified as primary products of the destruction of composite materials: stearic acid for polyurethane-based materials; imide of dithiocarbonic acid and 1-nonadecen in variants with ultrathene; and tetraaminopyrimidine and isocyanatodecan in variants with polyamide. N,N-dimethyldodecan amide, 2-methyloximundecanon and 2-nonacosane were identified for composites on the base of Lentex A4-1. Allyl methyl sulfide and imide of dithiocarbonic acid were found in variants with the samples of composites based on polyvinyl alcohol. The identified primary products of the destruction of composite materials belong to nontoxic compounds.

Stability of a biogas-producing bacterial, archaeal and fungal community degrading food residues.

PubMed

Bengelsdorf, Frank R; Gerischer, Ulrike; Langer, Susanne; Zak, Manuel; Kazda, Marian

2013-04-01

The resident microbiota was analyzed in a mesophilic, continuously operating biogas plant predominantly utilizing food residues, stale bread, and other waste cosubstrates together with pig manure and maize silage. The dominating bacterial, archaeal, and eukaryotic community members were characterized by two different 16S/18S rRNA gene culture-independent approaches. Prokaryotic 16S rRNA gene and eukaryotic 18S rRNA gene clone libraries were constructed and further analyzed by restriction fragment length polymorphism (RFLP), 16S/18S rRNA gene sequencing, and phylogenetic tree reconstruction. The most dominant bacteria belonged to the phyla Bacteriodetes, Chloroflexus, and Firmicutes. On the family level, the bacterial composition confirmed high differences among biogas plants studied so fare. In contrast, the methanogenic archaeal community was similar to that of other studied biogas plants. Furthermore, it was possible to identify fungi at the genus level, namely Saccharomyces and Mucor. Both genera, which are important for microbial degradation of complex compounds, were up to now not found in biogas plants. The results revealed their long-term presence as indicated by denaturating gradient gel electrophoresis (DGGE). The DGGE method confirmed that the main members of the microbial community were constantly present over more than one-year period. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Evaluation of Novel Broad-Range Real-Time PCR Assay for Rapid Detection of Human Pathogenic Fungi in Various Clinical Specimens▿

PubMed Central

Vollmer, Tanja; Störmer, Melanie; Kleesiek, Knut; Dreier, Jens

2008-01-01

In the present study, a novel broad-range real-time PCR was developed for the rapid detection of human pathogenic fungi. The assay targets a part of the 28S large-subunit ribosomal RNA (rDNA) gene. We investigated its application for the most important human pathogenic fungal genera, including Aspergillus, Candida, Cryptococcus, Mucor, Penicillium, Pichia, Microsporum, Trichophyton, and Scopulariopsis. Species were identified in PCR-positive reactions by direct DNA sequencing. A noncompetitive internal control was applied to prevent false-negative results due to PCR inhibition. The minimum detection limit for the PCR was determined to be one 28S rDNA copy per PCR, and the 95% detection limit was calculated to 15 copies per PCR. To assess the clinical applicability of the PCR method, intensive-care patients with artificial respiration and patients with infective endocarditis were investigated. For this purpose, 76 tracheal secretion samples and 70 heart valve tissues were analyzed in parallel by real-time PCR and cultivation. No discrepancies in results were observed between PCR analysis and cultivation methods. Furthermore, the application of the PCR method was investigated for other clinical specimens, including cervical swabs, nail and horny skin scrapings, and serum, blood, and urine samples. The combination of a broad-range real-time PCR and direct sequencing facilitates rapid screening for fungal infection in various clinical specimens. PMID:18385440

Evaluation of novel broad-range real-time PCR assay for rapid detection of human pathogenic fungi in various clinical specimens.

PubMed

Vollmer, Tanja; Störmer, Melanie; Kleesiek, Knut; Dreier, Jens

2008-06-01

In the present study, a novel broad-range real-time PCR was developed for the rapid detection of human pathogenic fungi. The assay targets a part of the 28S large-subunit ribosomal RNA (rDNA) gene. We investigated its application for the most important human pathogenic fungal genera, including Aspergillus, Candida, Cryptococcus, Mucor, Penicillium, Pichia, Microsporum, Trichophyton, and Scopulariopsis. Species were identified in PCR-positive reactions by direct DNA sequencing. A noncompetitive internal control was applied to prevent false-negative results due to PCR inhibition. The minimum detection limit for the PCR was determined to be one 28S rDNA copy per PCR, and the 95% detection limit was calculated to 15 copies per PCR. To assess the clinical applicability of the PCR method, intensive-care patients with artificial respiration and patients with infective endocarditis were investigated. For this purpose, 76 tracheal secretion samples and 70 heart valve tissues were analyzed in parallel by real-time PCR and cultivation. No discrepancies in results were observed between PCR analysis and cultivation methods. Furthermore, the application of the PCR method was investigated for other clinical specimens, including cervical swabs, nail and horny skin scrapings, and serum, blood, and urine samples. The combination of a broad-range real-time PCR and direct sequencing facilitates rapid screening for fungal infection in various clinical specimens.

The potentiality of cross-linked fungal chitosan to control water contamination through bioactive filtration.

PubMed

Tayel, Ahmed A; El-Tras, Wael F; Elguindy, Nihal M

2016-07-01

Water contamination, with heavy metals and microbial pathogens, is among the most dangerous challenges that confront human health worldwide. Chitosan is a bioactive biopolymer that could be produced from fungal mycelia to be utilized in various applied fields. An attempt to apply fungal chitosan for heavy metals chelation and microbial pathogens inhibition, in contaminated water, was performed in current study. Chitosan was produced from the mycelia of Aspergillus niger, Cunninghamella elegans, Mucor rouxii and from shrimp shells, using unified production conditions. The FT-IR spectra of produced chitosans were closely comparable. M. rouxii chitosan had the highest deacetylation degree (91.3%) and the lowest molecular weight (33.2kDa). All chitosan types had potent antibacterial activities against Escherichia coli and Staphylococcus aureus; the most forceful type was C. elegans chitosan. Chitosan beads were cross-linked with glutaraldehyde (GLA) and ethylene-glycol-diglycidyl ether (EGDE); linked beads became insoluble in water, acidic and alkaline solutions and could effectively adsorb heavy metals ions, e.g. copper, lead and zinc, in aqueous solution. The bioactive filter, loaded with EGDE- A. niger chitosan beads, was able to reduce heavy metals' concentration with >68%, and microbial load with >81%, after 6h of continuous water flow in the experimentally designed filter. Copyright © 2016 Elsevier B.V. All rights reserved.

Preliminary investigation into the prevalence of mucormycosis in the platypus (Ornithorhynchus anatinus) in three catchments in north-west Tasmania.

PubMed

Macgregor, J W; Holyoake, C S; Munks, S A; Robertson, I D; Warren, K S

2010-05-01

To investigate the distribution and prevalence of mucormycosis in platypus (Ornithorhynchus anatinus) from the Inglis, Emu and Black-Detention catchment areas in north-west Tasmania. A field study was performed and resulted in the examination of 44 wild platypuses; in addition, one dead platypus and two live platypuses were examined after they were independently submitted to a local veterinary clinic. No cases of mucormycosis were conclusively diagnosed. One platypus with signs consistent with those previously described in cases of mucormycosis was captured in the Emu River catchment. However, laboratory tests did not provide a definitive diagnosis for the lesion. Two platypuses from the Inglis catchment area had signs very similar to those previously described in cases of mucormycosis, but laboratory tests found Corynebacterium ulcerans to be the likely cause of the cutaneous ulcers on one of these platypuses and an unidentified fungal agent to be the cause of a cutaneous nodule in the other. These findings do not prove that mucormycosis is absent from the populations studied. However, they may indicate that the prevalence of disease is low. The possibility that Mucor amphibiorum is present in a subclinical form in platypuses, or infecting another reservoir, is not excluded. The findings also suggest that caution should be exercised when diagnosing mucormycosis based on clinical findings alone and raise the possibility that some cases may have been incorrectly diagnosed.

[Significance of MUC5B antibody in differential diagnosis between Aspergillus species and Mucorales of fungal sinusitis].

PubMed

Piao, Ying-shi; Liu, Hong-gang; Liu, Xian-jun

2008-04-01

To differentiate between Aspergillus species and Mucorales of fungal sinusitis by immunohistochemistry. Formalin-fixed paraffin-embedded tissue blocks of 66 cases of fungal sinusitis were retrieved from the archival files of Department of Pathology of Beijing Tongren Hospital during the period from 2001 to 2006. The samples included 29 cases of fungal balls, 12 cases of allergic fungal sinusitis, 24 cases of chronic invasive fungal sinusitis and 1 case of acute invasive fungal sinusitis. The types of fungi were 44 Aspergillus species (31 cases of A. fumigatus, 7 cases of A. flavus and 6 cases of A. terreus) and 22 Mucorales (14 cases of Mucor species and 8 cases of Rhizopus species). Immunohistochemistry was performed with MUC2, MUC5AC and MUC5B antibodies. The results were compared with histochemical study for periodic acid-Schiff (PAS) and Grocott methenamine silver (GMS) stains. Immunohistochemical study for MUC5B showed that the positive rate of Aspergillus species was 90.9%, in contrast to 4.5% in Mucorales (P < 0.001). The expression of MUC2 and MUC5AC was completely negative, whereas PAS and GMS stains were positive in all cases. MUC5B antibody appears to be a useful immunohistochemical marker for identifying fungal types in tissue sections, especially in distinguishing between Aspergillus species and Mucorales in fungal sinusitis.

Modeling the Effect of Modified Atmospheres on Conidial Germination of Fungi from Dairy Foods

PubMed Central

Nguyen Van Long, Nicolas; Vasseur, Valérie; Couvert, Olivier; Coroller, Louis; Burlot, Marion; Rigalma, Karim; Mounier, Jérôme

2017-01-01

Modified atmosphere packaging (MAP) is commonly applied to extend food shelf-life. Despite growth of a wide variety of fungal contaminants has been previously studied in relation to modified-atmospheres, few studies aimed at quantifying the effects of dioxygen (O2) and carbon dioxide (CO2) partial pressures on conidial germination in solid agar medium. In the present study, an original culture method was developed, allowing microscopic monitoring of conidial germination under modified-atmospheres in static conditions. An asymmetric model was utilized to describe germination kinetics of Paecilomyces niveus, Mucor lanceolatus, Penicillium brevicompactum, Penicillium expansum, and Penicillium roquefoti, using two main parameters, i.e., median germination time (τ) and maximum germination percentage (Pmax). These two parameters were subsequently modeled as a function of O2 partial pressure ranging from 0 to 21% and CO2 partial pressure ranging from 0.03 to 70% (8 tested levels for both O2 and CO2). Modified atmospheres with residual O2 or CO2 partial pressures below 1% and up to 70%, respectively, were not sufficient to totally inhibit conidial germination,. However, O2 levels < 1% or CO2 levels > 20% significantly increased τ and/or reduced Pmax, depending on the fungal species. Overall, the present method and results are of interest for predictive mycology applied to fungal spoilage of MAP food products. PMID:29163403

Disseminated fungal infection in two species of captive sharks.

PubMed

Marancik, David P; Berliner, Aimee L; Cavin, Julie M; Clauss, Tonya M; Dove, Alistair D M; Sutton, Deanna A; Wickes, Brian L; Camus, Alvin C

2011-12-01

In this report, two cases of systemic mycosis in captive sharks are characterized. These cases were progressive and ultimately culminated in terminal disease. Paecilomyces lilacinus, an uncommon pathogen in human and veterinary medicine, was associated with areas of necrosis in the liver, heart, and gill in a great hammerhead shark (Sphyrna mokarran). Fungal growth was observed from samples of kidney, spleen, spinal fluid, and coelomic cavity swabs. Dual fungal infection by Exophiala pisciphila and Mucor circinelloides was diagnosed in a juvenile zebra shark (Stegostoma fasciatum). Both fungi were present in the liver, with more severe tissue destruction associated with E. pisciphila. E. pisciphila also produced significant necrosis in the spleen and gill, while M. circinelloides was associated with only minimal tissue changes in the heart. Fungal cultures from liver, kidney, and spleen were positive for both E. pisciphila and M. circinelloides. Identification of P. lilacinus and M. circinelloides was based on colonial and hyphal morphology. E. pisciphila was identified by sequence analysis of the 28S rRNA D1/D2 region and the internal transcribed spacer (ITS) region between the 18S and 28S rRNA subunit. These cases, and a lack of information in the literature, highlight the need for further research and diagnostic sampling to further characterize the host-pathogen interaction between elasmobranchs and fungi.

Antimicrobial Applications of Transition Metal Complexes of Benzothiazole Based Terpolymer: Synthesis, Characterization, and Effect on Bacterial and Fungal Strains

PubMed Central

Riswan Ahamed, Mohamed A.; Azarudeen, Raja S.; Kani, N. Mujafar

2014-01-01

Terpolymer of 2-amino-6-nitro-benzothiazole-ethylenediamine-formaldehyde (BEF) has been synthesized and characterized by elemental analysis and various spectral techniques like FTIR, UV-Visible, and 1H and 13C-NMR. The terpolymer metal complexes were prepared with Cu2+, Ni2+, and Zn2+ metal ions using BEF terpolymer as a ligand. The complexes have been characterized by elemental analysis and IR, UV-Visible, ESR, 1H-NMR, and 13C-NMR spectral studies. Gel permeation chromatography was used to determine the molecular weight of the ligand. The surface features and crystalline behavior of the ligand and its complexes were analyzed by scanning electron microscope and X-ray diffraction methods. Thermogravimetric analysis was used to analyze the thermal stability of the ligand and its metal complexes. Kinetic parameters such as activation energy (E a) and order of reaction (n) and thermodynamic parameters, namely, ΔS, ΔF, S*, and Z, were calculated using Freeman-Carroll (FC), Sharp-Wentworth (SW), and Phadnis-Deshpande (PD) methods. Thermal degradation model of the terpolymer and its metal complexes was also proposed using PD method. Biological activities of the ligand and its complexes were tested against Shigella sonnei, Escherichia coli, Klebsiella species, Staphylococcus aureus, Bacillus subtilis, and Salmonella typhimurium bacteria and Aspergillus flavus, Aspergillus niger, Penicillium species, Candida albicans, Cryptococcus neoformans, Mucor species fungi. PMID:25298760

Atypical cause of forearm skin ulceration in a leukaemic child: mucormycosis. A case report.

PubMed

Zirak, C; Brutus, J P; De Mey, A

2005-01-01

Primary cutaneous mucormycosis is an uncommon, deep and aggressive fungal infection occurring mainly in immunosuppressed or diabetic patients. Rapid diagnosis and therapy are necessary to prevent a fatal outcome. An eight-year-old leukaemic child presented with a dark necrotic ulcer on the volar-ulnar aspect of the left forehand. The lesion had developed over seven days, beginning as a vasculo-haemorrhagic erythematous plaque. There was no known history of trauma to the area, but skin necrosis from external compression caused by an intravenous line could not be completely ruled out. The lesion rapidly progressed to a 5 x 8 cm painful necrotic ulcer with an erythematous border. Treatment with ichthyol dressing and intra-venous antibiotherapy failed to improve the condition. Wide debridement was performed and specimens were sent for microbiology and pathology examinations. Microscopic examination demonstrated broad, irregularly walled, non-septate fungal hyphae that were consistent with Mucor. Amphotericin B was administrated intravenously (1 mg/kg/day) and hydrogel and hydrophile adhesive polyurethane foam dressings were applied. After 12 days, the soft tissue defect was covered with a split-thickness skin graft, harvested from the lateral aspect of the thigh. Diagnosis of this infection is based on complete histopathological and microbiological studies. Awareness, and a high index of suspicion are required because of the potential fulminant and fatal course.

Microbiota Dynamics Associated with Environmental Conditions and Potential Roles of Cellulolytic Communities in Traditional Chinese Cereal Starter Solid-State Fermentation

PubMed Central

Li, Pan; Liang, Hebin; Lin, Wei-Tie; Feng, Feng

2015-01-01

Traditional Chinese solid-state fermented cereal starters contain highly complex microbial communities and enzymes. Very little is known, however, about the microbial dynamics related to environmental conditions, and cellulolytic communities have never been proposed to exist during cereal starter fermentation. In this study, we performed Illumina MiSeq sequencing combined with PCR-denaturing gradient gel electrophoresis to investigate microbiota, coupled with clone library construction to trace cellulolytic communities in both fermentation stages. A succession of microbial assemblages was observed during the fermentation of starters. Lactobacillales and Saccharomycetales dominated the initial stages, with a continuous decline in relative abundance. However, thermotolerant and drought-resistant Bacillales, Eurotiales, and Mucorales were considerably accelerated during the heating stages, and these organisms dominated until the end of fermentation. Enterobacteriales were consistently ubiquitous throughout the process. For the cellulolytic communities, only the genera Sanguibacter, Beutenbergia, Agrobacterium, and Erwinia dominated the initial fermentation stages. In contrast, stages at high incubation temperature induced the appearance and dominance of Bacillus, Aspergillus, and Mucor. The enzymatic dynamics of amylase and glucoamylase also showed a similar trend, with the activities clearly increased in the first 7 days and subsequently decreased until the end of fermentation. Furthermore, β-glucosidase activity continuously and significantly increased during the fermentation process. Evidently, cellulolytic potential can adapt to environmental conditions by changes in the community structure during the fermentation of starters. PMID:26002897

Lipase-catalyzed production of short-chain acids terpenyl esters of interest to the food industry.

PubMed

Laboret, F; Perraud, R

1999-12-01

The production of low molecular weight esters as flavor compounds by biotechnological processes has a potential interest for the food industry. The use of natural available substrates and enzymes is an essential part of the process design, because the products may obtain natural label. In this study, direct esterification of citronellol and geraniol with short-chain fatty acids catalyzed by free lipase from Mucor miehei was performed with high yields in n-hexane. The effects of the acid:alcohol ratio on the bioconversion rate of increasing chain length esters was investigated. To reach the optimum yield, substrates and enzyme concentration were determined. The inhibiting effects of acid are strongly attenuated by reducing the quantity of acid and increasing the amount of enzyme in media following the optimum values. Improvements have been made to increase the ester purity. The consumption of excess substrate by adding calculated amounts of acid gives a 10% yield enhancement, and leads to 100% pure terpenyl esters. The first steps to a scale-up application were attempted using a reactor that allowed us to produce ester quantities up to 100 cm3. Separation and purification of the products were treated with success, underlining the lipase stability and efficiency under the conditions of this study. The ability to recover the enzyme, and reusing it in bioconversions, plays a major role in reducing the cost of the overall process.

Treatment of Palm Oil Mill Effluent by a Microbial Consortium Developed from Compost Soils

PubMed Central

Nwuche, Charles O.; Ogbonna, James C.

2014-01-01

A method for the aerobic treatment of palm oil mill effluent (POME) was investigated in shake-flask experiments using a consortium developed from POME compost. POME was initially centrifuged at 4,000 g for 15 min and the supernatant was enriched with (NH4)2SO4 (0.5%) and yeast extract (0.25%) to boost its nitrogen content. At optimum pH (pH 4) and temperature (40°C) conditions, the chemical oxygen demand (COD) of the effluent decreased from 10,350 to 1,000 mg/L (90.3%) after 7 days. The total bacterial population determined by plate count enumeration was 2.4 × 106 CFU/mL, while the fungal count was 1.8 × 103 colonies/mL. Bacteria of the genera Pseudomonas, Flavobacterium, Micrococcus, and Bacillus were isolated, while the fungal genera included Aspergillus, Penicillium, Trichoderma, and Mucor. When the isolated species were each inoculated into separate batches of the raw effluent, both pH and COD were unchanged. However, at 75 and 50% POME dilutions, the COD dropped by 52 and 44%, respectively, while the pH increased from 4 to 7.53. POME treatment by aerobic method is sustainable and holds promising prospects for cushioning the environment from the problems associated with the use of anaerobic systems. PMID:27433536

Variations in the bioavailability of polycyclic aromatic hydrocarbons in industrial and agricultural soils after bioremediation.

PubMed

Guo, Meixia; Gong, Zongqiang; Allinson, Graeme; Tai, Peidong; Miao, Renhui; Li, Xiaojun; Jia, Chunyun; Zhuang, Jie

2016-02-01

The aim of this study was to demonstrate the variations in bioavailability remaining in industrial and agricultural soils contaminated by polycyclic aromatic hydrocarbons (PAHs) after bioremediation. After inoculation of Mycobacterium sp. and Mucor sp., PAH biodegradation was tested on a manufactured gas plant (MGP) soil and an agricultural soil. PAH bioavailability was assessed before and after biodegradation using solid-phase extraction (Tenax-TA extraction) and solid-phase micro-extraction (SPME) to represent bioaccessibility and chemical activity of PAHs, respectively. Only 3- and 4-ring PAHs were noticeably biodegradable in the MGP soil. PAH biodegradation in the agricultural soil was different from that in the MGP soil. The rapidly desorbing fractions (F(rap)) extracted by Tenax-TA and the freely dissolved concentrations of 3- and 4-ring PAHs determined by SPME from the MGP soil decreased after 30 days biodegradation; those values of the 5- and 6-ring PAHs changed to a lesser degree. For the agricultural soil, the F(rap) values of the 3- and 4-ring PAHs also decreased after the biodegradation experiment. The Tenax-TA extraction and the SPME have the potential to assess variations in the bioavailability of PAHs and the degree of biodegradation in contaminated MGP soils. In addition, Tenax-TA extraction is more sensitive than SPME when used in the agricultural soil. Copyright © 2015 Elsevier Ltd. All rights reserved.

Development and Evaluation of an Immunodiffusion Test for Diagnosis of Systemic Zygomycosis (Mucormycosis): Preliminary Report

PubMed Central

Jones, Kenneth W.; Kaufman, Leo

1978-01-01

An antigen analysis with filtrate and homogenate precipitinogens of single isolates of the zygomycetes Absidia corymbifera, Mucor pusillus, Rhizopus arrhizus, and Rhizopus oryzae demonstrated the presence of common antigens among the three genera as well as antigens which permit their differentiation. Selected homogenate antigens were valuable in developing a diagnostic immunodiffusion (ID) test for systemic zygomycosis. When sera from 43 patients with various proven mycoses other than zygomycosis were tested against each of the antigens, none formed precipitin bands identical to those formed by A. cormybifera, M. pusillus, and the Rhizopus spp. rabbit reference antisera. Sera from 23 normal persons and 25 diabetics did not react with any of the antigens. Homogenate antigens detected antibody in 8 of the 11 sera (73%) from suspected or proven cases of zygomycosis, whereas ID tests with filtrate antigens detected antibody in only 2 of the 11 sera (18%). Of the eight sera that reacted with the homogenate antigens, five only reacted with a specific Rhizopus sp. antigen, two only reacted with a specific M. pusillus antigen, and one only reacted with a specific A. corymbifera antigen. Study results show the ID test with homogenate antigens to be more specific and sensitive than the ID test with filtrate antigens and indicate that the former is a promising technique for diagnosing human zygomycosis. Images PMID:75212

Cnidarian-derived filamentous fungi from Brazil: isolation, characterisation and RBBR decolourisation screening.

PubMed

Da Silva, M; Passarini, M R Z; Bonugli, R C; Sette, L D

2008-12-01

Marine-derived fungi represent a valuable source of structurally novel and biologically active metabolites of industrial interest. They also have drawn attention for their capacity to degrade several pollutants, including textile dyes, organochlorides and polycyclic aromatic hydrocarbons (PAHs), among others. The fungal tolerance to higher concentrations of salt might be considered an advantage for bioremediation processes in the marine environment. Therefore, filamentous fungi were isolated from cnidarians (scleractinian coral and zoanthids) collected from the north coast of São Paulo State, Brazil. A total of 144 filamentous fungi were morphologically and molecularly characterised. Among them there were several species of Penicillium and Aspergillus, in addition to Cladosporium spp., Eutypella sp., Fusarium spp., Khuskia sp., Mucor sp., Peacilomyces sp., Phoma sp. and Trichoderma spp. These fungi were tested regarding their decolourisation activity for Remazol Brilliant Blue R (RBBR), a textile dye used as an initial screening for PAH-degrading fungi. The most efficient fungi for RBBR decolourisation after 12 days were Penicillium citrinum CBMAI 853 (100%), Aspergillus sulphureus CBMAI 849 (95%), Cladosporium cladosporioides CBMAI 857 (93%) and Trichoderma sp. CBMAI 852 (89%). Besides its efficiency for dye decolourisation within liquid media, C. cladosporioides CBMAI 857 also decolourised dye on solid media, forming a decolourisation halo. Further research on the biotechnological potential, including studies on PAH metabolism, of these selected fungi are in progress.

Evaluation of aerial microbial pollutants in Al-Haram Al-Nabawi during pilgrimage of 2013.

PubMed

Alananbeh, Kholoud M; Boquellah, Nahla; Kaff, Nadia Al; Ahmadi, Majid Al

2017-01-01

Al-Madinah Al-Munawwarah is the second holiest site in Islam. The possibility of new emerging microbes is valid due to the increased number of pilgrims. The objectives of the current study were to estimate the numbers of fungi and bacteria inside and outside Al-Haram Al-Nabawi and to find whether new bacterial and fungal species have emerged compared to previous studies. Air samples were collected twice a day from 12 spots and four directions during the pilgrim year of 2013 for four consecutive weeks by using the sedimentation method. Thirty five genera and fifty eight species were identified. The most recovered bacterial genera were Staphylococcus, Micrococcus, Bacillus, and Dermacoccus with 32.47%, 18.18%, 12.85%, and 11.23%, respectively. Fifty nine isolates of fungi were molecularly identified. Aspergillus species had the highest percentage (78%). The other fungal genera identified ( Alternaria triticina , Emericella nidulans , Emericella striata , Mucor circinelloides , Penicillium chrysogenum , Penicillium minioluteum , Rhizopus arrhizus , Rhizopus oryzae , and Syncephalastrum racemosum ) had less than 5% frequency. In places such as Al-Haram Al-Nabawi, a large and crowded public (millions) exist especially during pilgrimages and Ramadan, thus, exposure to microorganisms is high. On the other hand, microorganism infectivity depends on many factors including their virulence, landing site, and person's immunity. For those reasons, many aspects should be considered to avoid aerosol contaminants.

Production of bioethanol by direct bioconversion of oil-palm industrial effluent in a stirred-tank bioreactor.

PubMed

Alam, Md Zahangir; Kabbashi, Nassereldeen A; Hussin, S Nahdatul I S

2009-06-01

The purpose of this study was to evaluate the feasibility of producing bioethanol from palm-oil mill effluent generated by the oil-palm industries through direct bioconversion process. The bioethanol production was carried out through the treatment of compatible mixed cultures such as Thrichoderma harzianum, Phanerochaete chrysosporium, Mucor hiemalis, and yeast, Saccharomyces cerevisiae. Simultaneous inoculation of T. harzianum and S. cerevisiae was found to be the mixed culture that yielded the highest ethanol production (4% v/v or 31.6 g/l). Statistical optimization was carried out to determine the operating conditions of the stirred-tank bioreactor for maximum bioethanol production by a two-level fractional factorial design with a single central point. The factors involved were oxygen saturation level (pO(2)%), temperature, and pH. A polynomial regression model was developed using the experimental data including the linear, quadratic, and interaction effects. Statistical analysis showed that the maximum ethanol production of 4.6% (v/v) or 36.3 g/l was achieved at a temperature of 32 degrees C, pH of 6, and pO(2) of 30%. The results of the model validation test under the developed optimum process conditions indicated that the maximum production was increased from 4.6% (v/v) to 6.5% (v/v) or 51.3 g/l with 89.1% chemical-oxygen-demand removal.

Plant community influences on soil microfungal assemblages in boreal mixed-wood forests.

PubMed

De Bellis, T; Kernaghan, G; Widden, P

2007-01-01

We studied the relationships between assemblages of soil microfungi and plant communities in the southern boreal mixed-wood forests of Quebec. Sampling took place in 18,100 m2 plots from an existing research site. Plots were separated into three categories based on dominant overstory tree species: (i) trembling aspen, (ii) white birch and (iii) a mixture of white spruce and balsam fir. Within each plot a 1 m2 subplot was established in which the understory herbaceous layer was surveyed and soil cores were collected. Microfungi were isolated from soil cores with the soil-washing technique and isolates were identified morphologically. To support our morphological identifications DNA sequences were obtained for the most abundant microfungi. The most frequently occurring microfungal species were Penicillium thomii, P. spinulosum, P. janthinellum, Penicillium sp., P. melinii, Trichoderma polysporum, T. viride, T. hamatum, Mortierella ramanniana, Geomyces pannorum, Cylindrocarpon didymum, Mortierella sp. and Mucor hiemalis. Multivariate analyses (redundancy analysis followed by variance partitioning) revealed that most of the variation in microfungal communities was explained by understory plant species composition as opposed to soil chemistry or overstory tree species. In this floristically diverse system saprophytic microfungal assemblages were not correlated with the overstory tree species but were significantly correlated with the main understory herbs, thereby reflecting differences at a smaller spatial scale.

Outbreak of Invasive Wound Mucormycosis in a Burn Unit Due to Multiple Strains of Mucor circinelloides f. circinelloides Resolved by Whole-Genome Sequencing

PubMed Central

2018-01-01

ABSTRACT Mucorales are ubiquitous environmental molds responsible for mucormycosis in diabetic, immunocompromised, and severely burned patients. Small outbreaks of invasive wound mucormycosis (IWM) have already been reported in burn units without extensive microbiological investigations. We faced an outbreak of IWM in our center and investigated the clinical isolates with whole-genome sequencing (WGS) analysis. We analyzed M. circinelloides isolates from patients in our burn unit (BU1, Hôpital Saint-Louis, Paris, France) together with nonoutbreak isolates from Burn Unit 2 (BU2, Paris area) and from France over a 2-year period (2013 to 2015). A total of 21 isolates, including 14 isolates from six BU1 patients, were analyzed by whole-genome sequencing (WGS). Phylogenetic classification based on de novo assembly and assembly free approaches showed that the clinical isolates clustered in four highly divergent clades. Clade 1 contained at least one of the strains from the six epidemiologically linked BU1 patients. The clinical isolates were specific to each patient. Two patients were infected with more than two strains from different clades, suggesting that an environmental reservoir of clonally unrelated isolates was the source of contamination. Only two patients from BU1 shared one strain, which could correspond to direct transmission or contamination with the same environmental source. In conclusion, WGS of several isolates per patients coupled with precise epidemiological data revealed a complex situation combining potential cross-transmission between patients and multiple contaminations with a heterogeneous pool of strains from a cryptic environmental reservoir. PMID:29691339

Molybdenum accumulation, tolerance and molybdenum-selenium-sulfur interactions in Astragalus selenium hyperaccumulator and nonaccumulator species.

PubMed

DeTar, Rachael Ann; Alford, Élan R; Pilon-Smits, Elizabeth A H

2015-07-01

Some species hyperaccumulate selenium (Se) upwards of 0.1% of dry weight. This study addressed whether Se hyperaccumulators also accumulate and tolerate more molybdenum (Mo). A field survey revealed on average 2-fold higher Mo levels in three hyperaccumulator Astragali compared to three nonaccumulator Astragali, which were not significantly different. Next, a controlled study was performed where hyperaccumulators Astragalus racemosus and Astragalus bisulcatus were compared with nonaccumulators Astragalus drummondii and Astragalus convallarius for Mo accumulation and tolerance, alone or in the presence of Se. When grown on agar media with 0, 12, 24 or 48 mg L(-1) molybdate and/or 0, 1.6 or 3.2 mg L(-1) selenate, all species decreased in biomass with increasing Mo supply. Selenium did not impact biomass at the supplied levels. All Astragali accumulated Mo upwards of 0.1% of dry weight. Selenium levels were up to 0.08% in Astragalus racemosus and 0.04% Se in the other species. Overall, there was no correlation between Se hyperaccumulation and Mo accumulation capacity. However, the hyperaccumulators and nonaccumulators differed in some respects. While none of the species had a higher tissue Mo to sulfur (S) ratio than the growth medium, nonaccumulators had a higher Mo/S ratio than hyperaccumulators. Also, while molybdate and selenate reduced S accumulation in nonaccumulators, it did not in hyperaccumulators. Furthermore, A. racemosus had a higher Se/S ratio than its medium, while the other species did not. Additionally, Mo and Se treatment affected S levels in nonaccumulators, but not in hyperaccumulators. In conclusion, there is no evidence of a link between Se and Mo accumulation and tolerance in Astragalus. Sulfate transporters in hyperaccumulating Astragali appear to have higher sulfate specificity over other oxyanions, compared to nonaccumulators, and A. racemosus may have a transporter with enhanced selenate specificity relative to sulfate or molybdate. Copyright

High temperature effect on microflora of radish root-inhabited zone and nutrient solutions for radish growth

NASA Astrophysics Data System (ADS)

Borodina, E. V.; Tirranen, L. S.

The effect of high temperatures (35 and 45 °C) on microflora of the root zone of radish plants grown in phytotron was evaluated by the response of microorganisms from 9 indicator groups. Phytotron air temperature elevated to 35 °C for 20 hours caused no significant changes in qualitative and quantitative composition of the root microflora in experimental plants. By the end of the experiment, the species diversity of microflora had changed. The amount of phytopathogenic microorganisms decreased which can be interpreted as more stable co-existence of microflora with plants. The numbers of microbes from other indicator groups was in dynamic equilibrium. The plants' condition did not deteriorate either. Exposure to the temperature of 45 °C for 7 hours have been found to change the numbers and species diversity in the radish root zone microflora. The microorganisms were observed to increase their total numbers at the expense of certain indicator groups. Bacteria increased spore forms at the stage of spores. Colon bacillus bacteria of increased their numbers by the end of experiment by an order. By the end of experiment the roots of experiment plants had microscopic fungi from Mucor, Aspergillus, Trichoderma, Cladosporium genera. The observed changes in the microbial complex seem to be associated with the changes of root emissions and general deterioration of the plants' condition. It is suggested that the response of the microorganisms can be indicative of the condition of plants under investigation.

Microbiological quality of air in free-range and box-stall stable horse keeping systems.

PubMed

Wolny-Koładka, Katarzyna

2018-04-07

The aim of this study was to assess the microbiological quality of air in three horse riding centers differing in the horse keeping systems. The air samples were collected in one facility with free-range horse keeping system and two with box stalls of different sizes. The samples were collected over a period of 3 years (2015-2017), four times per year (spring, summer, autumn, winter) to assess the effect of seasonal changes. The prevalence of aerobic mesophilic bacteria, mold fungi, actinomycetes, Staphylococcus spp., and Escherichia coli was determined by the air collision method on Petri dishes with appropriate microbiological media. At the same time, air temperature, relative humidity, and particulate matter concentration (PM 10 , PM 2.5 ) were measured. It was found that the horse keeping system affects the occurrence of the examined airborne microorganisms. Over the 3-year period of study, higher temperature and humidity, as well as particulate matter concentration-which notoriously exceeded limit values-were observed in the facilities with the box-stall system. The air sampled from the largest horse riding center, with the largest number of horses and the box-stall system of horse keeping, was also characterized by the heaviest microbiological contamination. Among others, bacteria from the following genera: Staphylococcus spp., Streptococcus spp., Bacillus spp., and E. coli and fungi from the genera Aspergillus, Fusarium, Mucor, Rhizopus, Penicillium, Trichothecium, Cladosporium, and Alternaria were identified in the analyzed samples.

ESolvent-free, enzyme-catalyzed biodiesel production from mango, neem, and shea oils via response surface methodology.

PubMed

Nde, Divine Bup; Astete, Carlos; Boldor, Dorin

2015-12-01

Mango, neem and shea kernels produce non-conventional oils whose potentials are not fully exploited. To give an added value to these oils, they were transesterified into biodiesel in a solvent-free system using immobilized enzyme lipozyme from Mucor miehei. The Doehlert experimental design was used to evaluate the methyl ester (ME) yields as influenced by enzyme concentration-EC, temperature-T, added water content-AWC, and reaction time-RT. Biodiesel yields were quantified by (1)H NMR spectroscopy and subsequently modeled by a second order polynomial equation with interactions. Lipozyme enzymes were more tolerant to high temperatures in neem and shea oils reaction media compared to that of mango oil. The optimum reaction conditions EC, T, AWC, and RT assuring near complete conversion were as follows: mango oil 7.25 %, 36.6 °C, 10.9 %, 36.4 h; neem oil EC = 7.19 %, T = 45.7 °C, AWC = 8.43 %, RT = 25.08 h; and shea oil EC = 4.43 %, T = 45.65 °C, AWC = 6.21 % and RT = 25.08 h. Validation experiments of these optimum conditions gave ME yields of 98.1 ± 1.0, 98.5 ± 1.6 and 99.3 ± 0.4 % for mango, neem and shea oils, respectively, which all met ASTM biodiesel standards.

Microbiological, Nutritional, and Sensory Quality of Bread Produced from Wheat and Potato Flour Blends

PubMed Central

Ijah, Udeme Joshua Josiah; Aduloju, Mercy Oluwayemisi; Aransiola, Sesan Abiodun

2014-01-01

Dehydrated uncooked potato (Irish and sweet) flour was blended by weight with commercial wheat flour at 0 to 10% levels of substitution to make bread. Comparative study of the microbial and nutritional qualities of the bread was undertaken. The total aerobic bacterial counts ranged from 3.0 × 105 cfu/g to 1.09 × 106 cfu/g while the fungal counts ranged from 8.0 × 101 cfu/g to 1.20 × 103 cfu/g of the sample. Coliforms were not detected in the bread. Bacteria isolated were species of Bacillus, Staphylococcus, and Micrococcus while fungi isolates were species of Aspergillus, Penicillium, Rhizopus, and Mucor. The mean sensory scores (color, aroma, taste, texture, and general acceptability) were evaluated. The color of the bread baked from WF/IPF2 (wheat/Irish potato flour, 95 : 5%) blend was preferred to WF (wheat flour, 100%) while WF/SPF1 (wheat/sweet potato flour, 100%) and WF/IPF1 (wheat/Irish potato flour, 90 : 10%) aroma were preferred to WF. However, the bread baked from WF, WF/IPF2 (wheat flour/Irish potato flour, 95 : 5%), and WF/SPF2 (wheat/sweet potato flour, 95 : 5%) was more acceptable than other blends. The use of hydrated potato flour in bread making is advantageous due to increased nutritional value, higher bread yield, and reduced rate of staling. PMID:26904642

Studies on mould growth and biomass production using waste banana peel.

PubMed

Essien, J P; Akpan, E J; Essien, E P

2005-09-01

Hyphomycetous (Aspergillus fumigatus) and Phycomycetous (Mucor hiemalis) moulds were cultivated in vitro at room temperature (28 + 20 degrees C) to examined their growth and biomass production on waste banana peel agar (BPA) and broth (BPB) using commercial malt extract agar (MEA) and broth (MEB) as control. The moulds grew comparatively well on banana peel substrates. No significant difference (p > 0.05) in radial growth rates was observed between moulds cultivated on PBA and MEA, although growth rates on MEA were slightly better. Slight variations in sizes of asexual spores and reproductive hyphae were also observed between moulds grown on MEA and BPA. Smaller conidia and sporangiospores, and shorter aerial hyphae (conidiophores and sporangiophores) were noticed in moulds grown on BPA than on MEA. The biomass weight of the test moulds obtained after one month of incubation with BPB were only about 1.8 mg and 1.4 mg less than values recorded for A. fumigatus and M. hiemalis respectively, grown on MEB. The impressive performance of the moulds on banana peel substrate may be attributed to the rich nutrient (particularly the crude protein 7.8% and crude fat 11.6% contents) composition of banana peels. The value of this agricultural waste can therefore be increased by its use not only in the manufacture of mycological medium but also in the production of valuable microfungal biomass which is rich in protein and fatty acids.

QPCR detection of Mucorales DNA in bronchoalveolar lavage fluid to diagnose pulmonary mucormycosis.

PubMed

Scherer, Emeline E; Iriart, Xavier; Bellanger, Anne Pauline; Dupont, Damien; Guitard, Juliette; Gabriel, Frederic; Cassaing, Sophie; Charpentier, Eléna; Guenounou, Sarah; Cornet, Murielle; Botterel, Françoise; Rocchi, Steffi; Berceanu, Ana; Millon, Laurence

2018-06-06

Early diagnosis and treatment are essential to improving the outcome of mucormycosis. The aim of this retrospective study was to assess the contribution of quantitative PCR detection of Mucorales DNA in bronchoalveolar lavage fluids for early diagnosis of pulmonary mucormycosis.Bronchoalveolar lavage fluids (n=450) from 374 patients with pneumonia and immunosuppressive conditions were analyzed using a combination of 3 quantitative PCR assays targeting the main genera involved in mucormycosis in France ( Rhizomucor, Mucor/Rhizopus, Lichtheimia ).Among these 374 patients, 24 had at least one bronchoalveolar lavage with a positive PCR; 23/24 patients had radiological criteria for invasive fungal infections according to consensual criteria : 10 patients with probable or proven mucormycosis, and 13 additional patients with other invasive fungal infections (4 probable aspergillosis, 1 proven fusariosis, and 8 possible invasive fungal infections). Only 2/24 patients with a positive PCR on bronchoalveolar lavage had a positive Mucorales culture.PCR was also positive on serum in 17/24 patients. In most cases, PCR was first detected positive on sera (15/17). However, a positive PCR on bronchoalveolar lavage was the earliest and/or the only biological test revealing mucormycosis in 4 patients with a final diagnosis of probable or proven mucormycosis, 3 patients with probable aspergillosis and one patient with a possible invasive fungal infection.Mucorales PCR performed on bronchoalveolar lavage could provide additional arguments for earlier administration of Mucorales-directed antifungal therapy, thus improving the outcome of lung mucormycosis. Copyright © 2018 American Society for Microbiology.

Multicenter Evaluation of MIC Distributions for Epidemiologic Cutoff Value Definition To Detect Amphotericin B, Posaconazole, and Itraconazole Resistance among the Most Clinically Relevant Species of Mucorales

PubMed Central

Chakrabarti, A.; Chowdhary, A.; Cordoba, S.; Dannaoui, E.; Dufresne, P.; Fothergill, A.; Ghannoum, M.; Gonzalez, G. M.; Guarro, J.; Kidd, S.; Lass-Flörl, C.; Meis, J. F.; Pelaez, T.; Tortorano, A. M.; Turnidge, J.

2015-01-01

Clinical breakpoints (CBPs) have not been established for the Mucorales and any antifungal agent. In lieu of CBPs, epidemiologic cutoff values (ECVs) are proposed for amphotericin B, posaconazole, and itraconazole and four Mucorales species. Wild-type (WT) MIC distributions (organisms in a species-drug combination with no detectable acquired resistance mechanisms) were defined with available pooled CLSI MICs from 14 laboratories (Argentina, Australia, Canada, Europe, India, Mexico, and the United States) as follows: 10 Apophysomyces variabilis, 32 Cunninghamella bertholletiae, 136 Lichtheimia corymbifera, 10 Mucor indicus, 123 M. circinelloides, 19 M. ramosissimus, 349 Rhizopus arrhizus, 146 R. microsporus, 33 Rhizomucor pusillus, and 36 Syncephalastrum racemosum isolates. CLSI broth microdilution MICs were aggregated for the analyses. ECVs comprising ≥95% and ≥97.5% of the modeled populations were as follows: amphotericin B ECVs for L. corymbifera were 1 and 2 μg/ml, those for M. circinelloides were 1 and 2 μg/ml, those for R. arrhizus were 2 and 4 μg/ml, and those for R. microsporus were 2 and 2 μg/ml, respectively; posaconazole ECVs for L. corymbifera were 1 and 2, those for M. circinelloides were 4 and 4, those for R. arrhizus were 1 and 2, and those for R. microsporus were 1 and 2, respectively; both itraconazole ECVs for R. arrhizus were 2 μg/ml. ECVs may aid in detecting emerging resistance or isolates with reduced susceptibility (non-WT MICs) to the agents evaluated. PMID:25583714

Response of Soil Fungi Community Structure to Salt Vegetation Succession in the Yellow River Delta.

PubMed

Wang, Yan-Yun; Guo, Du-Fa

2016-10-01

High-throughput sequencing technology was used to reveal the composition and distribution of fungal community structure in the Yellow River Delta under bare land and four kinds of halophyte vegetation (saline seepweed, Angiospermae, Imperata and Apocynum venetum [A. venetum]). The results showed that the soil quality continuously improved with the succession of salt vegetation types. The soil fungi richness of mild-salt communities (Imperata and A. venetum) was relatively higher, with Shannon index values of 5.21 and 5.84, respectively. The soil fungi richness of severe-salt-tolerant communities (saline seepweed, Angiospermae) was relatively lower, with Shannon index values of 4.64 and 4.66, respectively. The UniFrac metric values ranged from 0.48 to 0.67 when the vegetation was in different succession stages. A total of 60,174 valid sequences were obtained for the five vegetation types, and they were classified into Ascomycota, Basidiomycota, Chytridiomycota, Glomeromycota and Mucoromycotina. Ascomycota had the greatest advantage among plant communities of Imperata and A. venetum, as indicated by relative abundances of 2.69 and 69.97 %, respectively. Basidiomycota had the greatest advantage among mild-salt communities of saline seepweed and Angiospermae, with relative abundances of 9.43 and 6.64 %, respectively. Soil physical and chemical properties were correlated with the distribution of the fungi, and Mucor was significantly correlated with soil moisture (r = 0.985; P < 0.01). Soil quality, salt vegetation and soil fungi were influenced by each other.

Reveromycins A and B from Streptomyces sp. 3–10: Antifungal Activity against Plant Pathogenic Fungi In vitro and in a Strawberry Food Model System

PubMed Central

Lyu, Ang; Liu, Hao; Che, Hongjie; Yang, Long; Zhang, Jing; Wu, Mingde; Chen, Weidong; Li, Guoqing

2017-01-01

This study was conducted to determine the antifungal activity of the metabolites from Streptomyces sp. 3–10, and to purify and identify the metabolites. Meanwhile, the taxonomic status of strain 3–10 was re-evaluated. The cultural filtrates of strain 3–10 in potato dextrose broth were extracted with ethyl acetate. The resulting crude extract at 1 and 5 μg/ml inhibited growth of 22 species in 18 genera of plant pathogenic fungi and Oomycetes, accounting for 92% of the total 24 tested species, suggesting that it has a wide antifungal spectrum. Two compounds were purified from the crude extract and were identified as reveromycins A and B, which demonstrated high antifungal activity against Botrytis cinerea, Mucor hiemails, Rhizopus stolonifer, and Sclerotinia sclerotiorum under acidic pH conditions. Both the crude extract and reveromycin A from strain 3–10 at 10, 50, and 100 μg/ml showed high efficacy in suppression of strawberry fruit rot caused by the above-mentioned four pathogens. The efficacy was comparable to that of corresponding commercial fungicides (pyrimethanil, captan, dimetachlone) used in management of these pathogens. Morphological, physiological, and phylogenetic characterization showed that strain 3–10 is closely related to Streptomyces yanglinensis 1307T, representing a novel phylotype in that species. This study reported a new strain with reveromycins-producing capability. The finding is important for further exploitation of reveromycins for agricultural use. PMID:28421050

Microbiota Dynamics Associated with Environmental Conditions and Potential Roles of Cellulolytic Communities in Traditional Chinese Cereal Starter Solid-State Fermentation.

PubMed

Li, Pan; Liang, Hebin; Lin, Wei-Tie; Feng, Feng; Luo, Lixin

2015-08-01

Traditional Chinese solid-state fermented cereal starters contain highly complex microbial communities and enzymes. Very little is known, however, about the microbial dynamics related to environmental conditions, and cellulolytic communities have never been proposed to exist during cereal starter fermentation. In this study, we performed Illumina MiSeq sequencing combined with PCR-denaturing gradient gel electrophoresis to investigate microbiota, coupled with clone library construction to trace cellulolytic communities in both fermentation stages. A succession of microbial assemblages was observed during the fermentation of starters. Lactobacillales and Saccharomycetales dominated the initial stages, with a continuous decline in relative abundance. However, thermotolerant and drought-resistant Bacillales, Eurotiales, and Mucorales were considerably accelerated during the heating stages, and these organisms dominated until the end of fermentation. Enterobacteriales were consistently ubiquitous throughout the process. For the cellulolytic communities, only the genera Sanguibacter, Beutenbergia, Agrobacterium, and Erwinia dominated the initial fermentation stages. In contrast, stages at high incubation temperature induced the appearance and dominance of Bacillus, Aspergillus, and Mucor. The enzymatic dynamics of amylase and glucoamylase also showed a similar trend, with the activities clearly increased in the first 7 days and subsequently decreased until the end of fermentation. Furthermore, β-glucosidase activity continuously and significantly increased during the fermentation process. Evidently, cellulolytic potential can adapt to environmental conditions by changes in the community structure during the fermentation of starters. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Genome sequence and transcriptome analyses of the thermophilic zygomycete fungus Rhizomucor miehei.

PubMed

Zhou, Peng; Zhang, Guoqiang; Chen, Shangwu; Jiang, Zhengqiang; Tang, Yanbin; Henrissat, Bernard; Yan, Qiaojuan; Yang, Shaoqing; Chen, Chin-Fu; Zhang, Bing; Du, Zhenglin

2014-04-21

The zygomycete fungi like Rhizomucor miehei have been extensively exploited for the production of various enzymes. As a thermophilic fungus, R. miehei is capable of growing at temperatures that approach the upper limits for all eukaryotes. To date, over hundreds of fungal genomes are publicly available. However, Zygomycetes have been rarely investigated both genetically and genomically. Here, we report the genome of R. miehei CAU432 to explore the thermostable enzymatic repertoire of this fungus. The assembled genome size is 27.6-million-base (Mb) with 10,345 predicted protein-coding genes. Even being thermophilic, the G + C contents of fungal whole genome (43.8%) and coding genes (47.4%) are less than 50%. Phylogenetically, R. miehei is more closerly related to Phycomyces blakesleeanus than to Mucor circinelloides and Rhizopus oryzae. The genome of R. miehei harbors a large number of genes encoding secreted proteases, which is consistent with the characteristics of R. miehei being a rich producer of proteases. The transcriptome profile of R. miehei showed that the genes responsible for degrading starch, glucan, protein and lipid were highly expressed. The genome information of R. miehei will facilitate future studies to better understand the mechanisms of fungal thermophilic adaptation and the exploring of the potential of R. miehei in industrial-scale production of thermostable enzymes. Based on the existence of a large repertoire of amylolytic, proteolytic and lipolytic genes in the genome, R. miehei has potential in the production of a variety of such enzymes.

Patterns of organic acids exuded by pioneering fungi from a glacier forefield are affected by carbohydrate sources

NASA Astrophysics Data System (ADS)

Brunner, Ivano; Goren, Asena; Schlumpf, Alessandro

2014-01-01

Bare soils in the area of retreating glaciers are ideal environments to study the role of microorganisms in the early soil formation and in processes of mineral weathering. The aim of our study was to investigate whether the source of carbohydrate would influence the patterns of organic acids exuded by fungal species. Three pioneering fungus species, isolated from fine granitic sediments in front of the Damma glacier from the central Swiss Alps, have previously been found to have the capability to exude organic acids and dissolve granite powder. In batch experiments, various carbohydrates, including glucose, cellulose, pectin, pollen, and cell remnants of cyanobacteria, fungi, and algae, were applied as carbohydrate sources and the patterns of exuded organic acids recorded. The results showed that two fungi, the zygomycete fungus Mucor hiemalis and the ascomycete fungus Penicillium chrysogenum, released a significantly higher amount of organic acids in dependence on specific carbohydrate sources. Pollen and algae as carbohydrate sources triggered significantly the exudation of malate in M. hiemalis, and pollen and cellulose that of oxalate in P. chrysogenum. We conclude that the occurrence of complex carbohydrate sources in nutrient-deficient deglaciated soils may positively influence the exudation of organic acids of fungi. In particular, pollen and remnants of other microorganisms can trigger the exudation of organic acids of fungi in order to promote the weathering of minerals and to make nutrients available that would otherwise be trapped in that cryospheric environment.

Dihydroxo-bridged dimeric Cu(II) system containing sandwiched non-coordinating phenylacetate anion: Crystal structure, spectroscopic, anti-bacterial, anti-fungal and DNA-binding studies of [(phen)(H2O)Cu(OH)2Cu(H2O)(phen)]2L.6H2O: (HL = phenylacetic acid; phen = 1,10-phenanthroline)

NASA Astrophysics Data System (ADS)

Iqbal, Muhammad; Ali, Saqib; Tahir, Muhammad Nawaz; Shah, Naseer Ali

2017-09-01

This paper reports the synthesis, X-ray crystal structure, DNA-binding, antibacterial and antifungal studies of a rare dihydroxo-bridged dinuclear copper(II) complex including 1,10-phenanthroline (Phen) ligands and phenylacetate (L) anions, [Cu2(Phen)2(OH)2(H2O)2].2L.6H2O. Structural data revealed distorted square-pyramidal geometry for each copper(II) atom with the basal plane formed by the two nitrogen atoms of the phenantroline ligand and the oxygen atoms of two bridging hydroxyl groups. The apical positions are filled by the oxygen atom from a water molecule. This forms a centrosymmetric cationic dimer where the uncoordinated phenylacetate ligands serve to balance the electrical charge. The dimers interact by means of hydrogen bonds aided by the coordinated as well as uncoordinated water molecules and phenyl-acetate moieties in the crystal lattice. The binding ability of the complex with salmon sperm DNA was determined using cyclic voltammetry and absorption spectroscopy yielding binding constants 2.426 × 104 M-1 and 1.399 × 104 M-1, respectively. The complex was screened against two Gram-positive (Micrococcus luteus and Bacillus subtilis) and one Gram-negative (Escherichia coli) bacterial strains exhibiting significant activity against all the three strains. The complex exhibited significant, moderate and no activity against fungal strains Mucor piriformis, Helminthosporium solani and Aspergillus Niger, respectively. These preliminary tests indicate the competence of the complex towards the development of a potent biological drug.

Composition and Variability of Biofouling Organisms in Seawater Reverse Osmosis Desalination Plants ▿ †

PubMed Central

Zhang, Minglu; Jiang, Sunny; Tanuwidjaja, Dian; Voutchkov, Nikolay; Hoek, Eric M. V.; Cai, Baoli

2011-01-01

Seawater reverse osmosis (SWRO) membrane biofouling remains a common challenge in the desalination industry, but the marine bacterial community that causes membrane fouling is poorly understood. Microbial communities at different stages of treatment processes (intake, cartridge filtration, and SWRO) of a desalination pilot plant were examined by both culture-based and culture-independent approaches. Bacterial isolates were identified to match the genera Shewanella, Alteromonas, Vibrio, and Cellulophaga based on 16S rRNA gene sequencing analysis. The 16S rRNA gene clone library of the SWRO membrane biofilm showed that a filamentous bacterium, Leucothrix mucor, which belongs to the gammaproteobacteria, accounted for nearly 30% of the clone library, while the rest of the microorganisms (61.2% of the total clones) were related to the alphaproteobacteria. 16S rRNA gene terminal restriction fragment length polymorphism (T-RFLP) analysis indicated that bacteria colonizing the SWRO membrane represented a subportion of microbes in the source seawater; however, they were quite different from those colonizing the cartridge filter. The examination of five SWRO membranes from desalination plants located in different parts of the world showed that although the bacterial communities from the membranes were not identical to each other, some dominant bacteria were commonly observed. In contrast, bacterial communities in source seawater were significantly different based on location and season. Microbial profiles from 14 cartridge filters collected from different plants also revealed spatial trends. PMID:21551282

In vitro production and antifungal activity of peptide ABP-dHC-cecropin A.

PubMed

Zhang, Jiaxin; Movahedi, Ali; Xu, Junjie; Wang, Mengyang; Wu, Xiaolong; Xu, Chen; Yin, Tongming; Zhuge, Qiang

2015-04-10

The antimicrobial peptide ABP-dHC-cecropin A is a small cationic peptide with potent activity against a wide range of bacterial species. Evidence of antifungal activity has also been suggested; however, testing of this peptide has been limited due to the low expression of cecropin proteins in Escherichia coli. To improve expression of this peptide in E. coli, ABP-dHC-cecropin A was cloned into a pSUMO vector and transformed into E. coli, resulting in the production of a pSUMO-ABP-dHC-cecropin A fusion protein. The soluble form of this protein was then purified by Ni-IDA chromatography, yielding a total of 496-mg protein per liter of fermentation culture. The SUMO-ABP-dHC-cecropin A fusion protein was then cleaved using a SUMO protease and re-purified by Ni-IDA chromatography, yielding a total of 158-mg recombinant ABP-dHC-cecropin A per liter of fermentation culture at a purity of ≥94%, the highest yield reported to date. Antifungal activity assays performed using this purified recombinant peptide revealed strong antifungal activity against both Candida albicans and Neurospora crassa, as well as Rhizopus, Fusarium, Alternaria, and Mucor species. Combined with previous analyses demonstrating strong antibacterial activity against a number of important bacterial pathogens, these results confirm the use of ABP-dHC-cecropin A as a broad-spectrum antimicrobial peptide, with significant therapeutic potential. Copyright © 2015 Elsevier B.V. All rights reserved.

Laboratory evaluation of Indian medicinal plants as repellents against malaria, dengue, and filariasis vector mosquitoes.

PubMed

Govindarajan, Marimuthu; Sivakumar, Rajamohan

2015-02-01

Mosquito-borne diseases have an economic impact, including loss in commercial and labor outputs, particularly in countries with tropical and subtropical climates; however, no part of the world is free from vector-borne diseases. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikungunya fever, West Nile virus, and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. In view of the recently increased interest in developing plant origin insecticides as an alternative to chemical insecticides, in the present study, the repellent activity of crude hexane, ethyl acetate, benzene, chloroform, and methanol extracts of leaf of Erythrina indica and root of Asparagus racemosus were assayed for their repellency against three important vector mosquitoes, viz., Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. The crude extract was applied on a membrane used for membrane feeding of unfed mosquitoes in a 1-ft cage. About 50 unfed 3-4-day-old laboratory-reared pathogen-free strains of A. stephensi, A. aegypti, and C. quinquefasciatus were introduced in a 1-ft cage fitted with a membrane with blood for feeding with temperature maintained at 37 °C through circulating water bath maintained at 40-45 °C. Three concentrations (1.0, 2.0, and 5.0 mg/cm(2)) of the crude extracts were evaluated. Repellents in E. indica afforded longer protection time against A. stephensi, A. aegypti, and C. quinquefasciatus than those in A. racemosus at 5.0 mg/cm(2) concentration, and the mean complete protection time ranged from 120 to 210 min with the different extracts tested. In this observation, these two plant crude extracts gave protection against mosquito bites; also, the repellent activity is dependent on the strength of the plant extracts. These results suggest that the leaf extract of E. indica and root extract of A. racemosus have the potential to

Ketopantoyl-lactone reductase from Candida parapsilosis: purification and characterization as a conjugated polyketone reductase.

PubMed

Hata, H; Shimizu, S; Hattori, S; Yamada, H

1989-02-24

Ketopantoyl-lactone reductase (2-dehydropantoyl-lactone reductase, EC 1.1.1.168) was purified and crystallized from cells of Candida parapsilosis IFO 0708. The enzyme was found to be homogeneous on ultracentrifugation, high-performance gel-permeation liquid chromatography and SDS-polyacrylamide gel electrophoresis. The relative molecular mass of the native and SDS-treated enzyme is approximately 40,000. The isoelectric point of the enzyme is 6.3. The enzyme was found to catalyze specifically the reduction of a variety of natural and unnatural polyketones and quinones other than ketopantoyl lactone in the presence of NADPH. Isatin and 5-methylisatin are rapidly reduced by the enzyme, the Km and Vmax values for isatin being 14 microM and 306 mumol/min per mg protein, respectively. Ketopantoyl lactone is also a good substrate (Km = 333 microM and Vmax = 481 mumol/min per mg protein). Reverse reaction was not detected with pantoyl lactone and NADP+. The enzyme is inhibited by quercetin, several polyketones and SH-reagents. 3,4-Dihydroxy-3-cyclobutene-1,2-dione, cyclohexenediol-1,2,3,4-tetraone and parabanic acid are uncompetitive inhibitors for the enzyme, the Ki values being 1.4, 0.2 and 3140 microM, respectively, with isatin as substrate. Comparison of the enzyme with the conjugated polyketone reductase of Mucor ambiguus (S. Shimizu, H. Hattori, H. Hata and H. Yamada (1988) Eur. J. Biochem. 174, 37-44) and ketopantoyl-lactone reductase of Saccharomyces cerevisiae suggested that ketopantoyl-lactone reductase is a kind of conjugated polyketone reductase.

Microbiology of processed edible insect products - Results of a preliminary survey.

PubMed

Grabowski, Nils Th; Klein, Günter

2017-02-21

Little is known of the microbiology of processed insect products. The present survey analysed a total of n=38 samples of deep-fried and spiced (Acheta domesticus, Locusta migratoria, and Omphisa fuscidentalis), cooked in soy sauce ("tsukudani"; Oxya yezoensis, Vespula flaviceps, and Bombyx mori), dried (A. domesticus, L. migatoria, Alphitobius diaperinus, Tenebrio molitor, B. mori, Hermetia illucens, and Musca domestica), powdered (H. illucens, T. molitor) and other (incl. deep-frozen B. mori and honeybee pollen) insect products microbiologically (total bacterial count [TBC], Enterobacteriaceae, staphylococci, bacilli, and yeasts and moulds counts, salmonellae, Listeria monocytogenes, and Escherichia coli). Although each product type revealed a microbiological profile of its own, dried and powdered insects ("class I") displayed markedly higher counts than the deep-fried and cooked ones ("class II"). Thresholds between class I and II products were estimated at 4.0 (TBC), 1.0 (Enterobacteriaceae, yeasts and moulds), 2.5 (staphylococci), and 3.0lgcfu/g (bacilli). All samples were negative for salmonellae, L. monocytogenes, E. coli and Stapyhlococcus aureus, but dried and powdered insects, as well as pollen, contained B. cereus, coliforms, Serratia liquefaciens, Listeria ivanovii, Mucor spp., Aspergillus spp., Penicillium spp., and Cryptococcus neoformans. Comparing the results with the hygiene criteria for edible insects proposed by Belgium and the Netherlands, class I products failed to comply with many bacterial count limits despite the absence of classical food pathogens. Therefore, class I products should always be consumed after another heating step as indicated by the manufacturer, until drying techniques are able to ensure lower bacterial counts. Copyright © 2016 Elsevier B.V. All rights reserved.

PUFA-induced cell death is mediated by Yca1p-dependent and -independent pathways, and is reduced by vitamin C in yeast.

PubMed

Johansson, Magnus; Chen, Xin; Milanova, Stefina; Santos, Cristiano; Petranovic, Dina

2016-03-01

Polyunsaturated fatty acids (PUFA) such as linoleic acid (LA, n-6, C18:2) and γ-linolenic acid (GLA, n-6, C18:3) are essential and must be obtained from the diet. There has been a growing interest in establishing a bio-sustainable production of PUFA in several microorganisms, e.g. in yeast Saccharomyces cerevisiae. However, PUFAs can also be toxic to cells because of their susceptibility to peroxidation. Here we investigated the negative effects of LA and GLA production on S. cerevisiae by characterizing a strain expressing active Δ6 and Δ12 desaturases from the fungus Mucor rouxii. Previously, we showed that the PUFA-producing strain has low viability, down-regulated genes for oxidative stress response, and decreased proteasome activity. Here we show that the PUFA strain accumulates high levels of reactive oxygen species (ROS) and lipid peroxides, and accumulates damaged proteins. The PUFA strain also showed great increase in metacaspase Yca1p activity, suggesting cells could die by caspase-mediated cell death. When treated with antioxidant vitamin C, ROS, lipid peroxidation and protein carbonylation were greatly reduced, and the activity of the metacaspase was significantly decreased too, ultimately doubling the lifespan of the PUFA strain. When deleting YCA1, the caspase-like activity and the oxidative stress decreased and although the lifespan was slightly prolonged, the phenotype could not be fully reversed, pointing that Yca1p was not the main executor of cell death. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Fungal mycelium and decomposition of needle litter in three contrasting coniferous forests

NASA Astrophysics Data System (ADS)

Virzo De Santo, Amalia; Rutigliano, Flora Angela; Berg, Björn; Fioretto, Antonietta; Puppi, Gigliola; Alfani, Anna

2002-08-01

The fungal mycelium ingrowth and the rates of mass loss and respiration of needle litter of Pinus pinea, Pinus laricio, Pinus sylvestris, and Abies alba were investigated, in three coniferous forests, over a 3-year period by means of a composite set of incubations. In the early stages, the fungal flora of the decomposing needles was dominated by dematiaceous hyphomycetes and coelomycetes. Basidiomycetes reached a peak after 6 months on pine needles, but were absent from the N-rich needles of A. alba. Soil fungi ( Penicillium, Trichoderma, Absidia, Mucor sp. pl.) became most frequent in later stages. At the end of the study period, the total mycelium amount showed the lowest values in all pine needles incubated in the P. laricio forest and the highest ones in P. pinea needles incubated in the P. pinea forest. In all data sets, as in data for boreal forests examined for comparison, the concentration of litter fungal mycelium versus litter mass loss followed a common exponential model. However, in later stages, the amount of litter fungal mycelium was very close to that of the humus at the incubation site, thus supporting the hypothesis of a logistic growth pattern. Respiration rates of decomposing litters varied with season and decreased with litter age to values close to those of the humus at the incubation site. Respiration of water-saturated litter was negatively correlated with the total mycelium concentration, and this was consistent with the observation that in far-decomposed litter only a minor fraction of the total mycelium is alive.

Alternaria sp. MG1, a resveratrol-producing fungus: isolation, identification, and optimal cultivation conditions for resveratrol production.

PubMed

Shi, Junling; Zeng, Qin; Liu, Yanlin; Pan, Zhongli

2012-07-01

Due to its potential in preventing or slowing the occurrence of many diseases, resveratrol (3,5,4'-trihydroxystilbene) has attracted great research interest. The objective of this study was to identify microorganisms from selected plants that produce resveratrol and to optimize the conditions for resveratrol production. Endophytes from Merlot wine grapes (Vitis vinifera L. cv. Merlot), wild Vitis (Vitis quinquangularis Rehd.), and Japanese knotweed (Polygonum cuspidatum Siebold & Zucc.) were isolated, and their abilities to produce resveratrol were evaluated. A total of 65 isolates were obtained and 21 produced resveratrol (6-123 μg/L) in liquid culture. The resveratrol-producing isolates belonged to seven genera, Botryosphaeria, Penicillium, Cephalosporium, Aspergillus, Geotrichum, Mucor, and Alternaria. The resveratrol-producing capability decreased or was completely lost in most isolates after three rounds of subculture. It was found that only the strain Alternaria sp. MG1 (isolated from cob of Merlot using GA1 medium) had stable and high resveratrol-producing capability in all subcultures. During liquid cultivation of Alternaria sp. MG1 in potato dextrose medium, the synthesis of resveratrol began on the first day, increased to peak levels on day 7, and then decreased sharply thereafter. Cell growth increased during cultivation and reached a stable and high level of biomass after 5 days. The best fermentation conditions for resveratrol production in liquid cultures of Alternaria sp. MG1 were an inoculum size of 6 %, a medium volume of 125 mL in a 250-mL flask, a rotation speed of 101 rpm, and a temperature of 27 °C.

Fungal Allergy: Pattern of sensitization over the past 11 years.

PubMed

Fernández-Soto, R; Navarrete-Rodríguez, E M; Del-Rio-Navarro, B E; Sienra-Monge, J J Luis; Meneses-Sánchez, N A; Saucedo-Ramírez, O J

2018-05-05

Although the prevalence of sensitization to fungi is not precisely known, it can reach 50% in inner cities and has been identified as a risk factor in the development of asthma. Whereas the prevalence of allergic diseases is increasing, it is unclear whether the same occurs with sensitization to fungi. A retrospective study was performed at the "Hospital Infantil de México Federico Gómez". From skin tests taken between 2004 and 2015, information was gathered about Alternaria alternata, Aspergillus fumigatus, Candida albicans, Cladosporium herbarum, Mucor mucedo and Penicillium notatum. The participating patients were 2-18 years old, presented some type of allergic condition, and underwent immediate hypersensitivity tests to the fungi herein examined. Descriptive analysis and chi-squared distribution were used. Of the 8794 patients included in the study, 14% showed a negative result to the entire panel of environmental allergens. The remaining 7565 individuals displayed sensitization to at least one fungus, which most frequently was Aspergillus, with a rate of 16.8%. When the patients were divided into age groups, the same trend was observed. The highest percentage of sensitization (58%) toward at least one type of fungus was found in 2014, and the lowest percentage (49.8%) in 2008. The rate of sensitization to at least one type of fungus was presently over 50%, higher than that detected in other medical centers in Mexico. This rate was constant over the 11-year study, and Aspergillus exhibited the greatest frequency of sensitization among the patients. Copyright © 2018 SEICAP. Published by Elsevier España, S.L.U. All rights reserved.

Diversity of microflora in the gut and casts of tropical composting earthworms reared on different substrates.

PubMed

Parthasarathi, K; Ranganathan, L S; Anandi, V; Zeyer, Josef

2007-01-01

The diversity of fungi, bacteria, yeast, actinomycetes and protozoa were analysed in the gut and casts of Eudrilus eugeniae, Lampito mauritii, Eisenia fetida and Perionyx excavatus, both qualitatively and quantitatively as influenced by different feed substrates like clay loam soil, cowdung and pressmud. While actinomycetes (Streptomyces albus, S. somaliensis, Nocardia asteroides, N. caviae and Saccharomonosporia) were not digested by any of these species of worms, protozoa (Amoeba proteus, A. terricola, Paramecium trichium, Euglena viridis, E. orientalis, Vorticella picta and Trichomonas hominis) and yeast (Candida tropicalis, C. krusei C. albicans and Cryptococcus neoformans) were totally digested. Certain species of fungi (Saksenae vasiformis, Mucor plumbeus, Cladosporium carrionii, C. herbacium, Alternaria sp., Cunninghamella echinulata, Mycetia sterila, Syncephalostrum racemosum, Curvalaria lunata, C. geniculata and Geotrichum candidum) and bacteria (Pseudomonas aeruginosa, Bacterium antitratum, Mima polymorpha, Enterobacter aerogenes, E. cloacae, Proteus vulgaris, P. mirabilis, P. rettgeri, Escherichia coli, Staphylococus citreus, Bacillus subtilis, B. cereus, Enterococci and Micrococci) were completely digested. Certain other species were not digested fungi like Aspergillus fumigatus, A. flavus, A. ochraceous, Trichoderma koningii (except by Eeugeniae), Fusarium moniliforme (except by E. eugeniae) and Rhizopus sp., and bacteria like Klebsiella pneumoniae and Morganella morganii) and these were multiplied during the transit of the organic residues through the gut of worms. The microbial proliferation was more in the casts, due to the environment prevailing--rich in nutrient supply and large surface area available for growth and reproduction of the microbes that lead to enhanced microbial activity and humic acid contents in the casts.

Biodiversity of antifungal lactic acid bacteria isolated from raw milk samples from cow, ewe and goat over one-year period.

PubMed

Delavenne, E; Mounier, J; Déniel, F; Barbier, G; Le Blay, G

2012-04-16

Antifungal lactic acid bacteria (ALAB) biodiversity was evaluated in raw milk from ewe, cow and goat over one year period. Lactic acid bacteria were enumerated using 8 semi-selective media, and systematically screened for their antifungal activity against 4 spoilage fungi commonly encountered in dairy products. Depending on the selective medium, between 0.05% (Elliker agar) and 5.5% (LAMVAB agar) screened colonies showed an antifungal activity. The great majority of these active colonies originated from cow (49%) and goat (43%) milks, whereas only 8% were isolated from ewe milk. Penicillium expansum was the most frequently inhibited fungus with 48.5% of colonies active against P. expansum among the 1235 isolated, followed by Mucor plumbeus with 30.6% of active colonies, Kluyveromyces lactis with only 12.1% of active colonies and Pichia anomala with 8.7% of active colonies. In the tested conditions, 94% of the sequenced active colonies belonged to Lactobacillus. Among them, targeted fungal species differed according to the Lactobacillus group, whose presence largely depended on year period and milk origin. The Lb. casei and Lb. reuteri groups, predominantly recovered in summer/fall, were overrepresented in the population targeting M. plumbeus, whereas isolates from the Lb. plantarum group, predominantly recovered in spring, were overrepresented in the population targeting K. lactis, the ones belonging to the Lb. buchneri group, predominantly recovered in spring, were overrepresented in the population targeting P. anomala. Raw milk, especially cow and goat milks from the summer/fall period appeared to be a productive reservoir for antifungal lactobacilli. Copyright © 2012 Elsevier B.V. All rights reserved.

Effects of inhaled fine dust on lung tissue changes and antibody response induced by spores of opportunistic fungi in goats.

PubMed

Purdy, Charles W; Layton, Robert C; Straus, David C; Ayers, J R

2008-04-01

To investigate the effects of sterile fine dust aerosol inhalation on antibody responses and lung tissue changes induced by Mucor ramosissimus or Trichoderma viride spores following intratracheal inoculation in goats. 36 weanling Boer-Spanish goats. 6 goats were allocated to each of 2 M ramosissimus-inoculated groups, 2 T viride-inoculated groups, and 2 control (tent or pen) groups. One of each pair of sporetreated groups and the tent control group were exposed 7 times to sterilized fine feedyard dust (mean+/-SD particle diameter, <7.72+/-0.69 microm) for 4 hours in a specially constructed tent. Goats in the 4 fungal treatment groups were inoculated intratracheally 5 times with a fungal spore preparation (30 mL), whereas tent control goats were intratracheally inoculated with physiologic saline (0.9% NaCl) solution (30 mL). Pen control goats were not inoculated or exposed to dust. Goats received an IV challenge with equine RBCs to assess antibody responses to foreign antigens. Postmortem examinations were performed at study completion (day 68) to evaluate lung tissue lesions. 5 of 7 deaths occurred between days 18 and 45 and were attributed to fine dust exposures prior to fungal treatments. Fine dust inhalation induced similar lung lesions and precipitating antibodies among spore-treated goats. Following spore inoculations, dust-exposed goats had significantly more spores per gram of consolidated lung tissue than did their nonexposed counterparts. Fine dust inhalation appeared to decrease the ability of goats to successfully clear fungal spores from the lungs following intratracheal inoculation.

Analysis of the influence of living environment and age on vaginal fungal microbiome in giant pandas (Ailuropoda melanoleuca) by high throughput sequencing.

PubMed

Chen, Danyu; Li, Caiwu; Feng, Lan; Zhang, Zhizhong; Zhang, Heming; Cheng, Guangyang; Li, Desheng; Zhang, Guiquan; Wang, Hongning; Chen, Yanxi; Feng, Mingfu; Wang, Chengdong; Wu, Honglin; Deng, Linhua; Ming, He; Yang, Xin

2018-02-01

A recent study has described the normal vaginal bacterial community in giant pandas, but there is a lack of knowledge of the fungal community residing in the vagina of giant pandas. In order to comprehensively understand the vaginal fungal microbial diversity and abundance in giant pandas, high throughput sequencing was used to analyse the ITS1 region, based on thirteen samples taken from the pandas' vaginas, which were grouped by sampling points and age. The results showed that the most abundant phyla were Basidiomycota (73.37%), followed by Ascomycota (20.04%), Zygomycota (5.23%), Glomeromycota (0.014%) and Chytridiomycota (0.006%). At the genus level, Guehomyces (37.92%) was the most abundant, followed by Cladosporium (9.072%), Trichosporon (6.2%) and Mucor (4.97%). Furthermore, Candida only accounted for a low percentage of the vaginal fungal community. With the saturation of rarefaction curves and fungal diversity indices, the samples from Dujiangyan and Chungking Safari Park (DC group) showed a higher fungal species richness and diversity than other living environments. Shannon diversity indices showed significant difference between group WL (Wolong nature reserve) and DC (P < .05). Additionally, a higher diversity was found in ten to fifteen years old (Group 2) than other groups. Group 2 and Group 3 displayed significant differences in the diversities of their vaginal fungal communities (P < .05). These data that has been collected from this research will be helpful for further study to improve the reproductive status of giant pandas. Copyright © 2018 Elsevier Ltd. All rights reserved.

Occurrence and species distribution of pathogenic Mucorales in unselected soil samples from France.

PubMed

Mousavi, B; Costa, J M; Arné, P; Guillot, J; Chermette, R; Botterel, F; Dannaoui, E

2018-04-01

Mucormycosis is a life-threatening invasive fungal disease that affects a variety of patient groups. Although Mucorales are mostly opportunistic pathogens originating from soil or decaying vegetation, there are currently few data on prevalence of this group of fungi in the environment. The aim of the present study was to assess the prevalence and diversity of species of Mucorales from soil samples collected in France. Two grams of soil were homogenized in sterile saline and plated on Sabouraud dextrose agar and RPMI agar supplemented with itraconazole or voriconazole. Both media contained chloramphenicol and gentamicin. The plates were incubated at 35 ± 2 °C and checked daily for fungal growth for a maximum of 7 d. Mucorales were subcultured for purity. Each isolate was identified phenotypically and molecular identification was performed by ITS sequencing. A total of 170 soil samples were analyzed. Forty-one isolates of Mucorales were retrieved from 38 culture-positive samples. Among the recovered isolates, 27 Rhizopus arrhizus, 11 Mucor circinelloides, one Lichtheimia corymbifera, one Rhizopus microsporus and one Cunninghamella bertholletiae were found. Positive soil samples came from cultivated fields but also from other types of soil such as flower beds. Mucorales were retrieved from samples obtained in different geographical regions of France. Voriconazole-containing medium improved the recovery of Mucorales compared with other media. The present study showed that pathogenic Mucorales are frequently recovered from soil samples in France. Species diversity should be further analyzed on a larger number of soil samples from different geographic areas in France and in other countries.

Multicenter evaluation of MIC distributions for epidemiologic cutoff value definition to detect amphotericin B, posaconazole, and itraconazole resistance among the most clinically relevant species of Mucorales.

PubMed

Espinel-Ingroff, A; Chakrabarti, A; Chowdhary, A; Cordoba, S; Dannaoui, E; Dufresne, P; Fothergill, A; Ghannoum, M; Gonzalez, G M; Guarro, J; Kidd, S; Lass-Flörl, C; Meis, J F; Pelaez, T; Tortorano, A M; Turnidge, J

2015-03-01

Clinical breakpoints (CBPs) have not been established for the Mucorales and any antifungal agent. In lieu of CBPs, epidemiologic cutoff values (ECVs) are proposed for amphotericin B, posaconazole, and itraconazole and four Mucorales species. Wild-type (WT) MIC distributions (organisms in a species-drug combination with no detectable acquired resistance mechanisms) were defined with available pooled CLSI MICs from 14 laboratories (Argentina, Australia, Canada, Europe, India, Mexico, and the United States) as follows: 10 Apophysomyces variabilis, 32 Cunninghamella bertholletiae, 136 Lichtheimia corymbifera, 10 Mucor indicus, 123 M. circinelloides, 19 M. ramosissimus, 349 Rhizopus arrhizus, 146 R. microsporus, 33 Rhizomucor pusillus, and 36 Syncephalastrum racemosum isolates. CLSI broth microdilution MICs were aggregated for the analyses. ECVs comprising ≥95% and ≥97.5% of the modeled populations were as follows: amphotericin B ECVs for L. corymbifera were 1 and 2 μg/ml, those for M. circinelloides were 1 and 2 μg/ml, those for R. arrhizus were 2 and 4 μg/ml, and those for R. microsporus were 2 and 2 μg/ml, respectively; posaconazole ECVs for L. corymbifera were 1 and 2, those for M. circinelloides were 4 and 4, those for R. arrhizus were 1 and 2, and those for R. microsporus were 1 and 2, respectively; both itraconazole ECVs for R. arrhizus were 2 μg/ml. ECVs may aid in detecting emerging resistance or isolates with reduced susceptibility (non-WT MICs) to the agents evaluated. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Expansion of signal transduction pathways in fungi by extensive genome duplication

PubMed Central

Corrochano, Luis M.; Kuo, Alan; Marcet-Houben, Marina; Polaino, Silvia; Salamov, Asaf; Villalobos-Escobedo, José M.; Grimwood, Jane; Álvarez, M. Isabel; Avalos, Javier; Bauer, Diane; Benito, Ernesto P.; Benoit, Isabelle; Burger, Gertraud; Camino, Lola P.; Cánovas, David; Cerdá-Olmedo, Enrique; Cheng, Jan-Fang; Domínguez, Angel; Eliáš, Marek; Eslava, Arturo P.; Glaser, Fabian; Gutiérrez, Gabriel; Heitman, Joseph; Henrissat, Bernard; Iturriaga, Enrique A.; Lang, B. Franz; Lavín, José L.; Lee, Soo Chan; Li, Wenjun; Lindquist, Erika; López-García, Sergio; Luque, Eva M.; Marcos, Ana T.; Martin, Joel; McCluskey, Kevin; Medina, Humberto R.; Miralles-Durán, Alejandro; Miyazaki, Atsushi; Muñoz-Torres, Elisa; Oguiza, José A.; Ohm, Robin A.; Orejas, Margarita; Ortiz-Castellanos, Lucila; Pisabarro, Antonio G.; Rodríguez-Romero, Julio; Ruiz-Herrera, José; Ruiz-Vázquez, Rosa; Sanz, Catalina; Schackwitz, Wendy; Shahriari, Mahdi; Shelest, Ekaterina; Silva-Franco, Fátima; Soanes, Darren; Syed, Khajamohiddin; Tagua, Víctor G.; Talbot, Nicholas J.; Thon, Michael R.; Tice, Hope; de Vries, Ronald P.; Wiebenga, Ad; Yadav, Jagjit S.; Braun, Edward L.; Baker, Scott E.; Garre, Victoriano; Schmutz, Jeremy; Horwitz, Benjamin A.; Torres-Martínez, Santiago; Idnurm, Alexander; Herrera-Estrella, Alfredo; Gabaldón, Toni; Grigoriev, Igor V.

2016-01-01

Summary Plants and fungi use light and other signals to regulate development, growth, and metabolism. The fruiting bodies of the fungus Phycomyces blakesleeanus are single cells that react to environmental cues, including light, but the mechanisms are largely unknown [1]. The related fungus Mucor circinelloides is an opportunistic human pathogen that changes its mode of growth upon receipt of signals from the environment to facilitate pathogenesis [2]. Understanding how these organisms respond to environmental cues should provide insights into the mechanisms of sensory perception and signal transduction by a single eukaryotic cell, and their role in pathogenesis. We sequenced the genomes of P. blakesleeanus and M. circinelloides, and show that they have been shaped by an extensive genome duplication or, most likely, a whole genome duplication (WGD), which is rarely observed in fungi [3-6]. We show that the genome duplication has expanded gene families, including those involved in signal transduction, and that duplicated genes have specialized, as evidenced by differences in their regulation by light. The transcriptional response to light varies with the developmental stage and is still observed in a photoreceptor mutant of P. blakesleeanus. A phototropic mutant of P. blakesleeanus with a heterozygous mutation in the photoreceptor gene madA demonstrates that photosensor dosage is important for the magnitude of signal transduction. We conclude that the genome duplication provided the means to improve signal transduction for enhanced perception of environmental signals. Our results will help to understand the role of genome dynamics in the evolution of sensory perception in eukaryotes. PMID:27238284

The interaction of fungi with the environment orchestrated by RNAi.

PubMed

Villalobos-Escobedo, José Manuel; Herrera-Estrella, Alfredo; Carreras-Villaseñor, Nohemí

2016-01-01

The fungal kingdom has been key in the investigation of the biogenesis and function of small RNAs (sRNAs). The discovery of phenomena such as quelling in Neurospora crassa represents pioneering work in the identification of the main elements of the RNA interference (RNAi) machinery. Recent discoveries in the regulatory mechanisms in some yeast and filamentous fungi are helping us reach a deeper understanding of the transcriptional and post-transcriptional gene-silencing mechanisms involved in genome protection against viral infections, DNA damage and transposon activity. Although most of these mechanisms are reasonably well understood, their role in the physiology, response to the environment and interaction of fungi with other organisms had remained elusive. Nevertheless, studies in fungi such as Mucor circinelloides, Magnaporthe oryzae, Cryptococcus neoformans, Trichoderma atroviride, Botrytis cinerea and others have started to shed light on the relevance of the RNAi pathway. In these fungi gene regulation by RNAi is important for growth, reproduction, control of viral infections and transposon activity, as well as in the development of antibiotic resistance and interactions with their hosts. Moreover, the increasing number of reports of the discovery of microRNA-like RNAs in fungi under different conditions highlights the importance of fungi as models for understanding adaptation to the environment using regulation by sRNAs. The goal of this review is to provide the reader with an up-to-date overview of the importance of RNAi in the interaction of fungi with their environment. © 2016 by The Mycological Society of America.

Comprehensive analysis of fungal diversity and enzyme activity in nuruk, a Korean fermenting starter, for acquiring useful fungi.

PubMed

Carroll, Emily; Trinh, Tran Ngoc; Son, Hokyoung; Lee, Yin-Won; Seo, Jeong-Ah

2017-05-01

Nuruk is a fermenting starter that is involved in the production of alcoholic beverages, and has been used in South Korea for a very long time. To analyze the fungal diversity, we collected a total of 59 nuruk samples from several companies and persons in 2013 to 2014, and obtained 364 isolates. All of the single isolated fungi were identified, both morphologically and molecularly, based on the sequences of ribosomal RNA gene [18S, ITS1-5.8S-ITS2, and 26S (D1/D2 region)]. In 46 nuruk samples out of 59 (78%), Saccharomycopsis fibuligera, a dimorphic yeast, was most frequently isolated. Among the filamentous fungi, Aspergillus and Lichtheimia were found in more than 50% of the samples with lower colony forming unit (CFU/g of sample) than those of yeasts. The yeasts S. fibuligera and Wickerhamomyces anomalus were counted with maximum 1.3-1.8 × 10 8 CFU/g. Among Mucorales fungi, Lichtheimia and Mucor were isolated in much higher numbers than Rhizopus and Rhizomucor. Overall, the home-made nuruks tend to contain more diverse filamentous fungi than the commercial nuruks. To acquire industrially useful filamentous fungi and yeasts, we analyzed the enzyme activities of α-amylase, glucoamylase and acid protease associated with brewing properties for 131 strains. Aspergillus oryzae and S. fibuligera had high α- and glucoamylase activities and most isolates of Lichtheimia ramosa had high acid protease activity. For further applications, 27 fungal strains were chosen based on isolation frequencies from nuruk, and the ability to produce useful enzyme.

Substrate specificity and kinetic properties of enzymes belonging to the hormone-sensitive lipase family: comparison with non-lipolytic and lipolytic carboxylesterases.

PubMed

Chahinian, Henri; Ali, Yassine Ben; Abousalham, Abdelkarim; Petry, Stefan; Mandrich, Luigi; Manco, Guiseppe; Canaan, Stephane; Sarda, Louis

2005-12-30

We have studied the kinetics of hydrolysis of triacylglycerols, vinyl esters and p-nitrophenyl butyrate by four carboxylesterases of the HSL family, namely recombinant human hormone-sensitive lipase (HSL), EST2 from Alicyclobacillus acidocaldarius, AFEST from Archeoglobus fulgidus, and protein RV1399C from Mycobacterium tuberculosis. The kinetic properties of enzymes of the HSL family have been compared to those of a series of lipolytic and non-lipolytic carboxylesterases including human pancreatic lipase, guinea pig pancreatic lipase related protein 2, lipases from Mucor miehei and Thermomyces lanuginosus, cutinase from Fusarium solani, LipA from Bacillus subtilis, porcine liver esterase and Esterase A from Aspergilus niger. Results indicate that human HSL, together with other lipolytic carboxylesterases, are active on short chain esters and hydrolyze water insoluble trioctanoin, vinyl laurate and olive oil, whereas the action of EST2, AFEST, protein RV1399C and non-lipolytic carboxylesterases is restricted to solutions of short chain substrates. Lipolytic and non-lipolytic carboxylesterases can be differentiated by their respective value of K(0.5) (apparent K(m)) for the hydrolysis of short chain esters. Among lipolytic enzymes, those possessing a lid domain display higher activity on tributyrin, trioctanoin and olive oil suggesting, then, that the lid structure contributes to enzyme binding to triacylglycerols. Progress reaction curves of the hydrolysis of p-nitrophenyl butyrate by lipolytic carboxylesterases with lid domain show a latency phase which is not observed with human HSL, non-lipolytic carboxylesterases, and lipolytic enzymes devoid of a lid structure as cutinase.

Bioconversion of 7-hydroxyflavanone: isolation, characterization and bioactivity evaluation of twenty-one phase I and phase II microbial metabolites.

PubMed

Mikell, Julie Rakel; Khan, Ikhlas Ahmad

2012-01-01

Microbial metabolism of 7-hydroxyflavanone (1) with fungal culture Cunninghamella blakesleeana (ATCC 8688a), yielded flavanone 7-sulfate (2), 7,4'-dihydroxyflavanone (3), 6,7-dihydroxyflavanone (4), 6-hydroxyflavanone 7-sulfate (5), and 7-hydroxyflavanone 6-sulfate (6). Mortierella zonata (ATCC 13309) also transformed 1 to metabolites 2 and 3 as well as 4'-hydroxyflavanone 7-sulfate (7), flavan-4-cis-ol 7-sulfate (8), 2',4'-dihydroxychalcone (9), 7,8-dihydroxyflavanone (10), 8-hydroxyflavanone 7-sulfate (11), and 8-methoxy-7-hydroxyflavanone (12). Beauveria bassiana (ATCC 7159) metabolized 1 to 2, 3, and 8, flavanone 7-O-β-D-O-4-methoxyglucopyranoside (13), and 8-hydroxyflavanone 7-O-β-D-O-4-methoxyglucopyranoside (14). Chaetomium cochlioides (ATCC 10195) also transformed 1 to 2, 3, 9, together with 7-hydroxy-4-cis-ol (15). Mucor ramannianus (ATCC 9628) metabolized 1 in addition to 7, to also 4,2',4'-trihydroxychalcone (16), 7,3',4'-trihydroxyflavanone (17), 4'-hydroxyflavanone 7-O-α-L-rhamnopyranoside (18), and 7,3',4'-trihydroxy-6-methoxyflavanone (19). The organism Aspergillus alliaceus (ATCC 10060) transformed 1 to metabolites 3, 16, 7,8,4'-trihydroxyflavanone (20), and 7-hydroxyflavanone 4'-sulfate (21). A metabolite of 1, flavanone 7-O-β-D-O-glucopyranoside (22) was produced by Rhizopus oryzae (ATCC 11145). Structures of the metabolic products were elucidated by means of spectroscopic data. None of the metabolites tested showed antibacterial, antifungal and antimalarial activities against selected organisms. Metabolites 4 and 16 showed weak antileishmanial activity.

Bacterial community analysis of an industrial wastewater treatment plant in Colombia with screening for lipid-degrading microorganisms.

PubMed

Silva-Bedoya, Lina Marcela; Sánchez-Pinzón, María Solange; Cadavid-Restrepo, Gloria Ester; Moreno-Herrera, Claudia Ximena

2016-11-01

The operation of wastewater treatment technologies depends on a combination of physical, chemical and biological factors. Microorganisms present in wastewater treatment plants play essential roles in the degradation and removal of organic waste and xenobiotic pollutants. Several microorganisms have been used in complementary treatments to process effluents rich in fats and oils. Microbial lipases have received significant industrial attention because of their stability, broad substrate specificity, high yields, and regular supply, as well as the fact that the microorganisms producing them grow rapidly on inexpensive media. In Colombia, bacterial community studies have focused on populations of cultivable nitrifying, heterotrophic and nitrogen-fixing bacteria present in constructed wetlands. In this study, culture-dependent methods, culture-independent methods (TTGE, RISA) and enzymatic methods were used to estimate bacterial diversity, to monitor temporal and spatial changes in bacterial communities, and to screen microorganisms that presented lipolytic activity. The dominant microorganisms in the Wastewater Treatment Plant (WWTP) examined in this study belonged to the phyla Firmicutes, Proteobacteria and Bacteroidetes. The enzymatic studies performed indicated that five bacterial isolates and three fungal isolates possessed the ability to degrade lipids; additionally, the Serratia, Kosakonia and Mucor genera presented lipase-mediated transesterification activity. The implications of these findings in regard to possible applications are discussed later in this paper. Our results indicate that there is a wide diversity of aerobic Gram-negative bacteria inhabiting the different sections of the WWTP, which could indicate its ecological condition, functioning and general efficiency. Copyright © 2016 Elsevier GmbH. All rights reserved.

Modelling the effect of water activity reduction by sodium chloride or glycerol on conidial germination and radial growth of filamentous fungi encountered in dairy foods.

PubMed

Nguyen Van Long, Nicolas; Rigalma, Karim; Coroller, Louis; Dadure, Robin; Debaets, Stella; Mounier, Jérôme; Vasseur, Valérie

2017-12-01

Water activity (a w ) is one of the most influential abiotic factors affecting fungal development in foods. The effects of a w reduction on conidial germination and radial growth are generally studied by supplementing culture medium with the non-ionic solute glycerol despite food a w can also depend on the concentration of ionic solutes such as sodium chloride (NaCl). The present study aimed at modelling and comparing the effects of a w , either modified using NaCl or glycerol, on radial growth and/or conidial germination parameters for five fungal species occurring in the dairy environment. The estimated cardinal values were then used for growth prediction and compared to growth kinetics observed on commercial fresh cheese. Overall, as compared to glycerol, NaCl significantly increased the fungistatic effect resulting from a w reduction by extending latency and/or reducing radial growth rates of Paecilomyces niveus, Penicillium brevicompactum, Penicillium expansum and Penicillium roqueforti but not of Mucor lanceolatus. Besides, NaCl significantly reduced a w range for conidial germination and delayed median germination time of P. expansum but not of P. roqueforti. Despite these observations, cardinal a w values obtained on glycerol-medium yielded similar predictions of radial growth and germination time in commercial fresh cheese as those obtained with NaCl. Thus, it indicates that, for the studied species and a w range used for model validation, the use of NaCl instead of glycerol as a a w depressor had only limited impact for fungal behavior prediction. Copyright © 2017 Elsevier Ltd. All rights reserved.

Mucorales from the semiarid of Pernambuco, Brazil

PubMed Central

de Azevedo Santiago, André Luiz Cabral Monteiro; dos Santos, Paulo Jorge Parreira; Maia, Leonor Costa

2013-01-01

Nineteen taxa of Mucorales, belonging to Absidia, Apophysomyces, Cunninghamella, Fennellomyces, Lichtheimia, Mucor, Mycotypha, Rhizopus and Syncephalastrum were isolated from 36 composite soil samples in three semiarid areas in the State of Pernambuco (Triunfo, Cabrobó and Belém de São Francisco), Northeast Brazil, which are characterized by Caatinga vegetation. Triunfo is preserved, whereas Cabroró and Belém de São Francisco are experiencing low and severe desertification processes, respectively. Mucorales were isolated in Petri dishes in triplicate from 5 mg samples of soil placed on the surface of wheat germ agar plus chloramphenicol and Cercobin [Dimethyl 4,49-(103 phenylene) bis (3-thioallophanate)] medium. The plates were left on a bench at room temperature (28 ± 2 °C) for 72 h of alternating dark and light periods. Absidia cylindrospora presented the highest amount of CFU/g of soil, followed by L. hyalospora, C. phaeospora and C. echinulata var. echinulata. The latter, and R. microsporus var. microsporus, presented the highest frequencies of occurrence. Soils from Triunfo showed higher diversity of Mucorales than the samples from the other areas, although without differing statistically in relation to species richness. The communities of Mucorales from the degraded areas were more similar, while that from the preserved area was quite different. Most of the identified specimens have been commonly isolated from soil in other Brazilian regions, which indicates that they are not endemic of the semiarid. Eleven taxa are registered for the first time in this ecosystem, while F. heterothallicus is reported for the first time in Brazil. PMID:24159320

Mucorales from the semiarid of Pernambuco, Brazil.

PubMed

de Azevedo Santiago, André Luiz Cabral Monteiro; Dos Santos, Paulo Jorge Parreira; Maia, Leonor Costa

2013-01-01

Nineteen taxa of Mucorales, belonging to Absidia, Apophysomyces, Cunninghamella, Fennellomyces, Lichtheimia, Mucor, Mycotypha, Rhizopus and Syncephalastrum were isolated from 36 composite soil samples in three semiarid areas in the State of Pernambuco (Triunfo, Cabrobó and Belém de São Francisco), Northeast Brazil, which are characterized by Caatinga vegetation. Triunfo is preserved, whereas Cabroró and Belém de São Francisco are experiencing low and severe desertification processes, respectively. Mucorales were isolated in Petri dishes in triplicate from 5 mg samples of soil placed on the surface of wheat germ agar plus chloramphenicol and Cercobin [Dimethyl 4,49-(103 phenylene) bis (3-thioallophanate)] medium. The plates were left on a bench at room temperature (28 ± 2 °C) for 72 h of alternating dark and light periods. Absidia cylindrospora presented the highest amount of CFU/g of soil, followed by L. hyalospora, C. phaeospora and C. echinulata var. echinulata. The latter, and R. microsporus var. microsporus, presented the highest frequencies of occurrence. Soils from Triunfo showed higher diversity of Mucorales than the samples from the other areas, although without differing statistically in relation to species richness. The communities of Mucorales from the degraded areas were more similar, while that from the preserved area was quite different. Most of the identified specimens have been commonly isolated from soil in other Brazilian regions, which indicates that they are not endemic of the semiarid. Eleven taxa are registered for the first time in this ecosystem, while F. heterothallicus is reported for the first time in Brazil.

Co-Production of Fungal Biomass Derived Constituents and Ethanol from Citrus Wastes Free Sugars without Auxiliary Nutrients in Airlift Bioreactor.

PubMed

Satari, Behzad; Karimi, Keikhosro; Taherzadeh, Mohammad J; Zamani, Akram

2016-02-26

The potential of two zygomycetes fungi, Mucor indicus and Rhizopus oryzae, in assimilating citrus waste free sugars (CWFS) and producing fungal chitosan, oil, and protein as well as ethanol was investigated. Extraction of free sugars from citrus waste can reduce its environmental impact by decreasing the possibility of wild microorganisms growth and formation of bad odors, a typical problem facing the citrus industries. A total sugar concentration of 25.1 g/L was obtained by water extraction of citrus waste at room temperature, used for fungal cultivation in shake flasks and airlift bioreactor with no additional nutrients. In shake flasks cultivations, the fungi were only able to assimilate glucose, while fructose remained almost intact. In contrast, the cultivation of M. indicus and R. oryzae in the four-liter airlift bioreactor resulted in the consumption of almost all sugars and production of 250 and 280 g fungal biomass per kg of consumed sugar, respectively. These biomasses correspondingly contained 40% and 51% protein and 9.8% and 4.4% oil. Furthermore, the fungal cell walls, obtained after removing the alkali soluble fraction of the fungi, contained 0.61 and 0.69 g chitin and chitosan per g of cell wall for M. indicus and R. oryzae, respectively. Moreover, the maximum ethanol yield of 36% and 18% was obtained from M. indicus and R. oryzae, respectively. Furthermore, that M. indicus grew as clump mycelia in the airlift bioreactor, while R. oryzae formed spherical suspended pellets, is a promising feature towards industrialization of the process.

Co-Production of Fungal Biomass Derived Constituents and Ethanol from Citrus Wastes Free Sugars without Auxiliary Nutrients in Airlift Bioreactor

PubMed Central

Satari, Behzad; Karimi, Keikhosro; Taherzadeh, Mohammad J.; Zamani, Akram

2016-01-01

The potential of two zygomycetes fungi, Mucor indicus and Rhizopus oryzae, in assimilating citrus waste free sugars (CWFS) and producing fungal chitosan, oil, and protein as well as ethanol was investigated. Extraction of free sugars from citrus waste can reduce its environmental impact by decreasing the possibility of wild microorganisms growth and formation of bad odors, a typical problem facing the citrus industries. A total sugar concentration of 25.1 g/L was obtained by water extraction of citrus waste at room temperature, used for fungal cultivation in shake flasks and airlift bioreactor with no additional nutrients. In shake flasks cultivations, the fungi were only able to assimilate glucose, while fructose remained almost intact. In contrast, the cultivation of M. indicus and R. oryzae in the four-liter airlift bioreactor resulted in the consumption of almost all sugars and production of 250 and 280 g fungal biomass per kg of consumed sugar, respectively. These biomasses correspondingly contained 40% and 51% protein and 9.8% and 4.4% oil. Furthermore, the fungal cell walls, obtained after removing the alkali soluble fraction of the fungi, contained 0.61 and 0.69 g chitin and chitosan per g of cell wall for M. indicus and R. oryzae, respectively. Moreover, the maximum ethanol yield of 36% and 18% was obtained from M. indicus and R. oryzae, respectively. Furthermore, that M. indicus grew as clump mycelia in the airlift bioreactor, while R. oryzae formed spherical suspended pellets, is a promising feature towards industrialization of the process. PMID:26927089

Mites and fungi in heavily infested stores in the Czech Republic.

PubMed

Hubert, J; Stejskal, V; Munzbergová, Z; Kubátová, A; Vánová, M; Zd'árková, E

2004-12-01

Toxigenic and allergen-producing fungi represent a serious hazard to human food and animal feed safety. Ninety-four fungal species were isolated from mite-infested samples of seeds taken from Czech seed stores. Fungi were isolated from the surface of four kinds of seeds (wheat, poppy, lettuce, and mustard) and from the gut and external surface of five species of mites (i.e., Acarus siro L., 1758, Caloglyphus rhizoglyphoides (Zachvatkin, 1973), Lepidoglyphus destructor (Schrank, 1781), Tyrophagus putrescentnae (Schrank, 1781) and Cheyletus malaccensis Oudemans 1903) separately. Multivariate analysis of fungi complex composition showed that the frequency of fungal was species significantly influenced by the kind of seed. Fungal frequencies differed between mites gut and exoskeleton surface and between the surfaces of mites and seeds. Three groups of fungal species were recognized: 1) mite surface-associated fungi: Penicillium brevicompactum, Alternaria alternata, and Aspergillus versicolor; 2) mite surface- and seed-associated fungi: Aspergillus niger, Penicillium crustosum, Penicillium aurantiogriseum, Penicillium chrysogenum, and Aspergillus flavus; and 3) seed-associated fungi: Cladosporium herbarum, Mucor dimorphosporus f. dimorphosporus, Botrytis cinerea, Penicillium griseofulvum, and Eurotium repens. Mite-carried species of microfungi are known to produce serious mycotoxins (e.g., aflatoxin B1, cyclopiazonic acid, sterigmatocystin, ochratoxin A, and nephrotoxic glycopeptides) as well as allergen producers (e.g., A. alternata and P. brevicompactum). Storage mites may play an important role in the spread of some medically hazardous micromycetes. In addition, these mite-fungi associations may heighten the risk of occurrence of mycotoxins in food and feed stuffs and cause mixed contamination by fungal and mite allergens.

Purification and characterisation of an antifungal protein, MCha-Pr, from the intercellular fluid of bitter gourd (Momordica charantia) leaves.

PubMed

Zhang, Beibei; Xie, Chengjian; Wei, Yunming; Li, Jing; Yang, Xingyong

2015-03-01

An antifungal protein, designated MCha-Pr, was isolated from the intercellular fluid of bitter gourd (Momordica charantia) leaves during a screen for potent antimicrobial proteins from plants. The isolation procedure involved a combination of extraction, ammonium sulphate precipitation, gel filtration on Bio-Gel P-6, ion exchange chromatography on CM-Sephadex, an additional gel filtration on HiLoad 16/60 Superdex 30, and finally, HPLC on a SOURCE 5RPC column. Matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry indicated that the protein had a molecular mass of 25733.46Da. Automated Edman degradation was used to determine the N-terminal sequence of MCha-Pr, and the amino acid sequence was identified as V-E-Y-T-I-T-G-N-A-G-N-T-P-G-G. The MCha-Pr protein has some similarity to the pathogenesis-related proteins from Atropa belladonna (deadly nightshade), Solanum tuberosum (potato), Ricinus communis (castor bean), and Nicotiana tabacum (tobacco). Analysis of the circular dichroism spectra indicated that MCha-Pr predominantly contains α-helix and β-sheet structures. MCha-Pr had inhibitory effects towards a variety of fungal species and the 50% inhibition of fungal growth (IC50) for Alternaria brassicae, Cercospora personata, Fusarium oxysporum, Mucor sp., and Rhizoctonia solani are 33 μM, 42 μM, 37 μM, 40 μM, and 48 μM, respectively. In addition, this antifungal protein can inhibit the germination of A. brassicae spores at 12.5 μM. These results suggest that MCha-Pr in bitter gourd leaves plays a protective role against phytopathogens and has a wide antimicrobial spectrum. Copyright © 2014 Elsevier Inc. All rights reserved.

Effect of two intermediate electron donors, NADPH and FADH(2), on Spirulina Delta (6)-desaturase co-expressed with two different immediate electron donors, cytochrome b (5) and ferredoxin, in Escherichia coli.

PubMed

Kurdrid, Pavinee; Subudhi, Sanjukta; Cheevadhanarak, Supapon; Tanticharoen, Morakot; Hongsthong, Apiradee

2007-12-01

When the gene desD encoding Spirulina Delta(6)-desaturase was heterologously expressed in E. coli, the enzyme was expressed without the ability to function. However, when this enzyme was co-expressed with an immediate electron donor, i.e. the cytochrome b (5) domain from Mucor rouxii, the results showed the production of GLA (gamma-linolenic acid), the product of the reaction catalyzed by Delta(6)-desaturase. The results revealed that in E. coli cells, where cytochrome b (5) is absent and ferredoxin, a natural electron donor of Delta(6)-desaturase, is present at a very low level, the cytochrome b (5) domain can complement for the function of ferredoxin in the host cells. In the present study, the Spirulina-ferredoxin gene was cloned and co-expressed with the Delta(6)-desaturase in E. coli. In comparison to the co-expression of cytochrome b ( 5 ) with the Delta(6)-desaturase, the co-expression with ferredoxin did not cause any differences in the GLA level. Moreover, the cultures containing the Delta(6)-desaturase co-expressed with cytochrome b (5) and ferredoxin were exogenously supplied with the intermediate electron donors, NADPH (nicotinamide adenine dinucleotide phosphate, reduced form) and FADH(2) (flavin adenine dinucleotide, reduced form), respectively. The GLA level in these host cells increased drastically, by approximately 50%, compared to the cells without the intermediate electron donors. The data indicated that besides the level of immediate electron donors, the level of intermediate electron donors is also critical for GLA production. Therefore, if the pools of the immediate and intermediate electron donors in the cells are manipulated, the GLA production in the heterologous host will be affected.

Studies on drug metabolism by fungi colonizing decomposing human cadavers. Part II: biotransformation of five model drugs by fungi isolated from post-mortem material.

PubMed

Martínez-Ramírez, Jorge A; Walther, Grit; Peters, Frank T

2015-04-01

The present study investigated the in vitro metabolic capacity of 28 fungal strains isolated from post-mortem material towards five model drugs: amitriptyline, metoprolol, mirtazapine, promethazine, and zolpidem. Each fungal strain was incubated at 25 °C for up to 120 h with each of the five models drugs. Cunninghamella elegans was used as positive control. Aliquots of the incubation mixture were centrifuged and 50 μL of the supernatants were diluted and directly analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) with product ion scanning. The remaining mixture was analyzed by full scan gas chromatography-mass spectrometry (GC-MS) after liquid-liquid extraction and acetylation. The metabolic activity was evaluated through the total number of detected metabolites (NDM) produced in each model and fungal strains and the percentage of parent drug remaining (%RPD) after up to five days of incubation. All the tested fungal strains were capable of forming mammalian phase I metabolites. Fungi from the normal fungal flora of the human body such as Candida sp., Geotrichum candidum, and Trichosporon asahii) formed up to seven metabolites at %RPD values greater than 52% but no new fungal metabolites (NFM). In contrast, some airborne fungal strains like Bjerkandera adusta, Chaetomium sp, Coriolopsis sp., Fusarium solani and Mucor plumbeus showed NDM values exceeding those of the positive control, complete metabolism of the parent drug in some models and formation of NFM. NFM (numbers in brackets) were detected in four of the five model drugs: amitriptyline (18), metoprolol (4), mirtazapine (8), and zolpidem (2). The latter NFM are potential candidates for marker substances indicating post-mortem fungal metabolism. Copyright © 2014 John Wiley & Sons, Ltd.

Fungal Diversity of Human Gut Microbiota Among Eutrophic, Overweight, and Obese Individuals Based on Aerobic Culture-Dependent Approach.

PubMed

Borges, Francis M; de Paula, Thaís O; Sarmiento, Marjorie R A; de Oliveira, Maycon G; Pereira, Maria L M; Toledo, Isabela V; Nascimento, Thiago C; Ferreira-Machado, Alessandra B; Silva, Vânia L; Diniz, Cláudio G

2018-06-01

Fungi have a complex role in the intestinal tract, influencing health and disease, with dysbiosis contributing to obesity. Our objectives were to investigate fungal diversity in human gut microbiota among eutrophic, overweight, and obese. Epidemiological and nutritional information were collected from adult individuals, as well as stool samples processed for selective fungi isolation and identification by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (yeasts) or microculture (filamentous fungi). Further 18S rDNA sequencing was performed to confirm identification. The mean count of fungi was 241 CFU/g of feces. Differences in the population level of the filamentous fungi were observed within eutrophic and obese groups. Overall, 34 genera were identified. The predominant phylum was Ascomycota with 20 different genera, followed by Basidiomycota and Zygomycota. As for Ascomycota, the most prevalent species were Paecilomyces sp., Penicillium sp., Candida sp., Aspergillus sp., Fonsecaea sp., and Geotrichum sp. (76.39, 65.28, 59.72, 58.33, 12.50, and 9.72%, respectively). As for Basidiomycota, Trichosporon sp. and Rhodotorula sp. were the most prevalent (30.56 and 15.28%, respectively), and for Zygomycota, Rhizopus sp. and Mucor sp. were the most numerous (15.28 and 9.72%, respectively). As expected there is a mycobiota shift towards obesity, with slightly higher diversity associated to eutrophic individuals. This mycobiota shift seems also to be related to the nutritional behavior of the individuals, as observed that the macronutrients intake may be positively related to the different fungi occurrences. Other studies are needed to better understand relationships between mycobiota and obesity, which could be used in future obesity treatments.

The Calcineurin Pathway Inhibitor Tacrolimus Enhances the In Vitro Activity of Azoles against Mucorales via Apoptosis

PubMed Central

Shirazi, F.

2013-01-01

The calcineurin pathway regulates antifungal drug resistance and the virulence of several major human-pathogenic fungi, including the recalcitrant Mucorales. We hypothesized that the fungistatic triazoles posaconazole (PCZ) and itraconazole (ICZ) become fungicidal in the setting of the calcineurin inhibitor tacrolimus (TCR) and that such an effect is mediated through apoptosis. Fungicidal activity and apoptosis were studied using standard microbiological techniques and hyphal metabolic and vital dye reduction assays at 37°C in RPMI 1640. Apoptosis was characterized by detecting intracellular Ca2+, phosphatidylserine (PS) externalization, DNA fragmentation, plasma membrane integrity, chromatin condensation, reactive oxygen species (ROS) generation, caspase-like activity, ATP, and cytochrome c release. MICs for PCZ and ICZ alone were significantly higher (8 to 128 μg/ml) than those of PCZ or ICZ plus TCR (0.25 to 4 μg/ml) for Rhizopus oryzae, Cunninghamella bertholletiae, and Mucor circinelloides. Both PCZ and ICZ in combination with TCR became fungicidal, and their activity was mediated through increased apoptotic cell death of R. oryzae (10 to 50%), C. bertholletiae (5 to 50%), and M. circinelloides (5 to 55%) germlings, with morphological apoptotic changes characterized by externalization of PS, nuclear condensation, and DNA fragmentation. Moreover, activation of the caspase-like activity was correlated with cell death induced by TCR plus PCZ or ICZ. These changes correlated with elevated intracellular Ca2+ and ROS levels and disturbance of mitochondrial potential. We found that PCZ or ICZ in combination with TCR renders Mucorales sensitive to triazoles via apoptotic death. These observations could serve as a new paradigm for the development of new therapeutic strategies. PMID:23851337

The calcineurin pathway inhibitor tacrolimus enhances the in vitro activity of azoles against Mucorales via apoptosis.

PubMed

Shirazi, F; Kontoyiannis, D P

2013-09-01

The calcineurin pathway regulates antifungal drug resistance and the virulence of several major human-pathogenic fungi, including the recalcitrant Mucorales. We hypothesized that the fungistatic triazoles posaconazole (PCZ) and itraconazole (ICZ) become fungicidal in the setting of the calcineurin inhibitor tacrolimus (TCR) and that such an effect is mediated through apoptosis. Fungicidal activity and apoptosis were studied using standard microbiological techniques and hyphal metabolic and vital dye reduction assays at 37°C in RPMI 1640. Apoptosis was characterized by detecting intracellular Ca(2+), phosphatidylserine (PS) externalization, DNA fragmentation, plasma membrane integrity, chromatin condensation, reactive oxygen species (ROS) generation, caspase-like activity, ATP, and cytochrome c release. MICs for PCZ and ICZ alone were significantly higher (8 to 128 μg/ml) than those of PCZ or ICZ plus TCR (0.25 to 4 μg/ml) for Rhizopus oryzae, Cunninghamella bertholletiae, and Mucor circinelloides. Both PCZ and ICZ in combination with TCR became fungicidal, and their activity was mediated through increased apoptotic cell death of R. oryzae (10 to 50%), C. bertholletiae (5 to 50%), and M. circinelloides (5 to 55%) germlings, with morphological apoptotic changes characterized by externalization of PS, nuclear condensation, and DNA fragmentation. Moreover, activation of the caspase-like activity was correlated with cell death induced by TCR plus PCZ or ICZ. These changes correlated with elevated intracellular Ca(2+) and ROS levels and disturbance of mitochondrial potential. We found that PCZ or ICZ in combination with TCR renders Mucorales sensitive to triazoles via apoptotic death. These observations could serve as a new paradigm for the development of new therapeutic strategies.

Signum, a new fungicide with interesting properties in resistance management of fungal diseases in strawberries.

PubMed

Hauke, K; Creemers, P; Brugmans, W; Van Laer, S

2004-01-01

Signum, a new fungicide developed by BASF, was applied during 6 successive years against fungal diseases in strawberries. The product is formulated as a water dispersible granule, containing 6.7 % pyraclostrobin and 26.7 % boscalid. Pyraclostrobin is similar in chemistry to other strobilurin fungicides like kresoxim-methyl and trifloxystrobin, registered for fruit disease control. Boscalid belongs to the class of carboxyanilides. Both components in the premix formulation combine two different biochemical modes of action in the fungal cell respiration. Therefore, this co-formulation gives a broad-spectrum activity and also a reduced resistance risk for different target pathogens. Botrytis cinerea is the most important disease on strawberry-fruits and thus the control of fruit rot is mainly focused on this fungus. In average over 6 years, Signum has not only given a very good control against Botrytis fruit rot, but it has also shown a high performance in the control of Colletotrichum. Besides, Signum provides good control of powdery mildew (Podosphaera aphanis) and limits the shift to other fruit rots like leather rot (Phytophthora cactorum and leak (Rhizopus, Mucor). The availability of several categories of fungicide families with a different mode of action gives opportunities in alternating different fungicides and is the best guarantee for a sustainable control of fruit rot in all kinds of strawberry production methods. Signum should be integrated in an overall disease management program. Trials, in which the applications of Signum were timed on disease forecasting, based on environmental factors favorable for Botrytis development, were very promising. This tool can also help in establishing the IPM-concept in the production of strawberries.

Production of ligninolytic enzymes and some diffusible antifungal compounds by white-rot fungi using potato solid wastes as the sole nutrient source.

PubMed

Schalchli, H; Hormazábal, E; Rubilar, O; Briceño, G; Mutis, A; Zocolo, G J; Diez, M C

2017-10-01

The aim of this study was to evaluate the synthesis of ligninolytic enzymes and some diffusible antifungal compounds by white-rot fungi (WRF) using peels or discarded potato as the sole nutrient source. The strain Trametes hirsuta Ru-513 highlighted for its laccase activity (595 ± 33 U l -1 ), which is able to decolourize 87% of an anthraquinone dye using potato peels as the sole nutritional support. A native polyacrylamide gel of laccase proteins showed the presence of two isoenzymes, corresponding to proteins of 56 and 67 kDa, which were detected by SDS-PAGE. The antifungal activity of ethyl acetate extracts was evaluated by the agar diffusion method, where Anthracophyllum discolor Sp4 and Inonotus sp. Sp2 showed the highest inhibition zones of Mucor miehei. The fungal extracts also inhibited Fusarium oxysporum and Botrytis cinerea growth, with inhibition zones of up to 18 mm. The extract with the highest antifungal activity, from A. discolor Sp4 grown in discarded potato medium, was analysed using a gas chromatograph coupled to a mass spectrometer. Among the identified compounds, chlorinated aromatic compounds and veratryl alcohol were the most abundant compounds. The results revealed the relevance of potato waste valorization for the sustainable production of ligninolytic enzymes and antifungal compounds. This study reports the synthesis of ligninolytic enzymes and diffusible antifungal compounds by WRF using potato wastes as the sole nutrient source and suggests a relationship between the enzymatic activity and the synthesis of antifungal compounds. These compounds and the synthesis of halogen compounds by WRF using agro-industrial wastes have been poorly studied before. © 2017 The Society for Applied Microbiology.

Fungal Diversity and Community Composition of Culturable Fungi in Stanhopea trigrina Cast Gibberellin Producers

PubMed Central

Salazar-Cerezo, Sonia; Martinez-Montiel, Nancy; Cruz-Lopez, Maria del Carmen; Martinez-Contreras, Rebeca D.

2018-01-01

Stanhopea tigrina is a Mexican endemic orchid reported as a threatened species. The naturally occurring microorganisms present in S. tigrina are unknown. In this work, we analyzed the diversity of endophytic and epiphytic culturable fungi in S. tigrina according to morphological and molecular identification. Using this combined approach, in this study we retrieved a total of 634 fungal isolates that presented filamentous growth, which were grouped in 134 morphotypes that were associated to 63 genera, showing that S. tigrina harbors a rich diversity of both endophytic and epiphytic fungi. Among these, the majority of the isolates corresponded to Ascomycetes, with Trichoderma and Penicillium as the most frequent genera followed by Fusarium and Aspergillus. Non-ascomycetes isolated were associated only to the genus Mucor (Mucoromycota) and Schizophyllum (Basidiomycota). Identified genera showed a differential distribution considering their epiphytic or endophytic origin, the tissue from which they were isolated, and the ability of the orchid to grow on different substrates. To our knowledge, this work constitutes the first study of the mycobiome of S. tigrina. Interestingly, 21 fungal isolates showed the ability to produce gibberellins. Almost half of the isolates were related to the gibberellin-producer genus Penicillium based on morphological and molecular identification. However, the rest of the isolates were related to the following genera, which have not been reported as gibberellin producers so far: Bionectria, Macrophoma, Nectria, Neopestalotiopsis, Talaromyces, Trichoderma, and Diplodia. Taken together, we found that S. tigrina possess a significant fungal diversity that could be a rich source of fungal metabolites with the potential to develop biotechnological approaches oriented to revert the threatened state of this orchid in the near future. PMID:29670591

Fungal Diversity and Community Composition of Culturable Fungi in Stanhopea trigrina Cast Gibberellin Producers.

PubMed

Salazar-Cerezo, Sonia; Martinez-Montiel, Nancy; Cruz-Lopez, Maria Del Carmen; Martinez-Contreras, Rebeca D

2018-01-01

Stanhopea tigrina is a Mexican endemic orchid reported as a threatened species. The naturally occurring microorganisms present in S. tigrina are unknown. In this work, we analyzed the diversity of endophytic and epiphytic culturable fungi in S. tigrina according to morphological and molecular identification. Using this combined approach, in this study we retrieved a total of 634 fungal isolates that presented filamentous growth, which were grouped in 134 morphotypes that were associated to 63 genera, showing that S. tigrina harbors a rich diversity of both endophytic and epiphytic fungi. Among these, the majority of the isolates corresponded to Ascomycetes, with Trichoderma and Penicillium as the most frequent genera followed by Fusarium and Aspergillus . Non-ascomycetes isolated were associated only to the genus Mucor (Mucoromycota) and Schizophyllum (Basidiomycota). Identified genera showed a differential distribution considering their epiphytic or endophytic origin, the tissue from which they were isolated, and the ability of the orchid to grow on different substrates. To our knowledge, this work constitutes the first study of the mycobiome of S. tigrina . Interestingly, 21 fungal isolates showed the ability to produce gibberellins. Almost half of the isolates were related to the gibberellin-producer genus Penicillium based on morphological and molecular identification. However, the rest of the isolates were related to the following genera, which have not been reported as gibberellin producers so far: Bionectria, Macrophoma, Nectria, Neopestalotiopsis, Talaromyces, Trichoderma , and Diplodia . Taken together, we found that S. tigrina possess a significant fungal diversity that could be a rich source of fungal metabolites with the potential to develop biotechnological approaches oriented to revert the threatened state of this orchid in the near future.

Hydroxylation of the herbicide isoproturon by fungi isolated from agricultural soil.

PubMed

Rønhede, Stig; Jensen, Bo; Rosendahl, Søren; Kragelund, Birthe B; Juhler, René K; Aamand, Jens

2005-12-01

Several asco-, basidio-, and zygomycetes isolated from an agricultural field were shown to be able to hydroxylate the phenylurea herbicide isoproturon [N-(4-isopropylphenyl)-N',N'-dimethylurea] to N-(4-(2-hydroxy-1-methylethyl)phenyl)-N',N'-dimethylurea and N-(4-(1-hydroxy-1-methylethyl)phenyl)-N',N'-dimethylurea. Bacterial metabolism of isoproturon has previously been shown to proceed by an initial demethylation to N-(4-isopropylphenyl)-N'-methylurea. In soils, however, hydroxylated metabolites have also been detected. In this study we identified fungi as organisms that potentially play a major role in the formation of these hydroxylated metabolites in soils treated with isoproturon. Isolates of Mortierella sp. strain Gr4, Phoma cf. eupyrena Gr61, and Alternaria sp. strain Gr174 hydroxylated isoproturon at the first position of the isopropyl side chain, yielding N-(4-(2-hydroxy-1-methylethyl)phenyl)-N',N'-dimethylurea, while Mucor sp. strain Gr22 hydroxylated the molecule at the second position, yielding N-(4-(1-hydroxy-1-methylethyl)phenyl)-N',N'-dimethylurea. Hydroxylation was the dominant mode of isoproturon transformation in these fungi, although some cultures also produced traces of the N-demethylated metabolite N-(4-isopropylphenyl)-N'-methylurea. A basidiomycete isolate produced a mixture of the two hydroxylated and N-demethylated metabolites at low concentrations. Clonostachys sp. strain Gr141 and putative Tetracladium sp. strain Gr57 did not hydroxylate isoproturon but N demethylated the compound to a minor extent. Mortierella sp. strain Gr4 also produced N-(4-(2-hydroxy-1-methylethyl)phenyl)-N'-methylurea, which is the product resulting from combined N demethylation and hydroxylation.

Hydroxylation of the Herbicide Isoproturon by Fungi Isolated from Agricultural Soil

PubMed Central

Rønhede, Stig; Jensen, Bo; Rosendahl, Søren; Kragelund, Birthe B.; Juhler, René K.; Aamand, Jens

2005-01-01

Several asco-, basidio-, and zygomycetes isolated from an agricultural field were shown to be able to hydroxylate the phenylurea herbicide isoproturon [N-(4-isopropylphenyl)-N′,N′-dimethylurea] to N-(4-(2-hydroxy-1-methylethyl)phenyl)-N′,N′-dimethylurea and N-(4-(1-hydroxy-1-methylethyl)phenyl)-N′,N′-dimethylurea. Bacterial metabolism of isoproturon has previously been shown to proceed by an initial demethylation to N-(4-isopropylphenyl)-N′-methylurea. In soils, however, hydroxylated metabolites have also been detected. In this study we identified fungi as organisms that potentially play a major role in the formation of these hydroxylated metabolites in soils treated with isoproturon. Isolates of Mortierella sp. strain Gr4, Phoma cf. eupyrena Gr61, and Alternaria sp. strain Gr174 hydroxylated isoproturon at the first position of the isopropyl side chain, yielding N-(4-(2-hydroxy-1-methylethyl)phenyl)-N′,N′-dimethylurea, while Mucor sp. strain Gr22 hydroxylated the molecule at the second position, yielding N-(4-(1-hydroxy-1-methylethyl)phenyl)-N′,N′-dimethylurea. Hydroxylation was the dominant mode of isoproturon transformation in these fungi, although some cultures also produced traces of the N-demethylated metabolite N-(4-isopropylphenyl)-N′-methylurea. A basidiomycete isolate produced a mixture of the two hydroxylated and N-demethylated metabolites at low concentrations. Clonostachys sp. strain Gr141 and putative Tetracladium sp. strain Gr57 did not hydroxylate isoproturon but N demethylated the compound to a minor extent. Mortierella sp. strain Gr4 also produced N-(4-(2-hydroxy-1-methylethyl)phenyl)-N′-methylurea, which is the product resulting from combined N demethylation and hydroxylation. PMID:16332769

Partial purification and characterization of exoinulinase from Kluyveromyces marxianus YS-1 for preparation of high-fructose syrup.

PubMed

Singh, Ram Sarup; Dhaliwal, Rajesh; Puri, Munish

2007-05-01

An extracellular exoinulinase (2,1-beta-D fructan fructanohydrolase, EC 3.2.1.7), which catalyzes the hydrolysis of inulin into fructose and glucose, was purified 23.5-fold by ethanol precipitation, followed by Sephadex G-100 gel permeation from a cell-free extract of Kluyveromyces marxianus YS-1. The partially purified enzyme exhibited considerable activity between pH 5 to 6, with an optimum pH of 5.5, while it remained stable (100%) for 3 h at the optimum temperature of 50 degrees C. Mn2+ and Ca2+ produced a 2.4-fold and 1.2-fold enhancement in enzyme activity, whereas Hg2+ and Ag2+ completely inhibited the inulinase. A preparation of the partially purified enzyme effectively hydrolyzed inulin, sucrose, and raffinose, yet no activity was found with starch, lactose, and maltose. The enzyme preparation was then successfully used to hydrolyze pure inulin and raw inulin from Asparagus racemosus for the preparation of a high-fructose syrup. In a batch system, the exoinulinase hydrolyzed 84.8% of the pure inulin and 86.7% of the raw Asparagus racemosus inulin, where fructose represented 43.6 mg/ml and 41.3 mg/ml, respectively.

Testing soil-like substrate for growing plants in bioregenerative life support systems

NASA Astrophysics Data System (ADS)

Gros, J. B.; Lasseur, Ch.; Tikhomirov, A. A.; Manukovsky, N. S.; Kovalev, V. S.; Ushakova, S. A.; Zolotukhin, I. G.; Tirranen, L. S.; Karnachuk, R. A.; Dorofeev, V. Yu.

We studied soil-like substrate (SLS) as a potential candidate for plant cultivation in bioregenerative life support systems (BLSS). The SLS was obtained by successive conversion of wheat straw by oyster mushrooms and worms. Mature SLS contained 9.5% humic acids and 4.9% fulvic acids. First, it was shown that wheat, bean and cucumber yields as well as radish yields when cultivated on mature SLS were comparable to yields obtained on a neutral substrate (expanded clay aggregate) under hydroponics. Second, the possibility of increasing wheat and radish yields on the SLS was assessed at three levels of light intensity: 690, 920 and 1150 μmol m -2 s -1 of photosynthetically active radiation (PAR). The highest wheat yield was obtained at 920 μmol m -2 s -1, while radish yield increased steadily with increasing light intensity. Third, long-term SLS fertility was tested in a BLSS model with mineral and organic matter recycling. Eight cycles of wheat and 13 cycles of radish cultivation were carried out on the SLS in the experimental system. Correlation coefficients between SLS nitrogen content and total wheat biomass and grain yield were 0.92 and 0.97, respectively, and correlation coefficients between nitrogen content and total radish biomass and edible root yield were 0.88 and 0.87, respectively. Changes in hormone content (auxins, gibberellins, cytokinins and abscisic acid) in the SLS during matter recycling did not reduce plant productivity. Quantitative and species compositions of the SLS and irrigation water microflora were also investigated. Microbial community analysis of the SLS showed bacteria from Bacillus, Pseudomonas, Proteus, Nocardia, Mycobacterium, Arthrobacter and Enterobacter genera, and fungi from Trichoderma, Penicillium, Fusarium, Aspergillus, Mucor, Botrytis, and Cladosporium genera.

Effect of Organic Dust Exposure on Pulmonary Functions in Workers of Vegetable Market with Special Reference to its Microbial Content.

PubMed

Goel, Arun; Omar, Balram Ji; Kathrotia, Rajesh; Patil, Prashant M; Mittal, Sunita

2018-01-01

Wholesale vegetable market is a rich source of generation of organic dust as loads of fruits and vegetables are loaded and unloaded here daily. Thus, regular workers are exposed to this organic dust for a considerable period of time depending on their work schedule. This study was planned to determine the microbial status of organic dust and to explore its association with pulmonary functions in the workers of wholesale vegetable market in Rishikesh. It was a cross-sectional exploratory/observational study. Thirty-five apparently healthy adult males were selected from vegetable market having no history of any chronic illness. Smokers and alcoholic were excluded from the study. The same number of age- and sex-matched controls with the same exclusion criteria were recruited from workers not working in the vegetable market and also not exposed to any other kinds of organic dust. Microbial culture of air in the vegetable market was done. It was compared with the microbial status of air in the working place of controls. Pulmonary functions of all the workers were performed with the help of digital spirometer (Helios 401). Bacterial and fungal concentration was found to be significantly higher in the air of vegetable market as compared to air in the workplace of controls (such as coagulase-negative staphylococci >25 colony-forming unit (CFU) at incubation temperature vs. 10-12 CFU at incubation temperature, significant growth of Mucor , Aspergillus niger , and Candida nonalbicans in vegetable market as compared to workplace of controls). Pulmonary function parameters (percentage forced expiratory volume in 1 st s (FEV 1 ), percentage predicted forced expiratory flow in mid-half of expiration, and FEV 1 ) of workers exposed to organic dust in vegetable market were also significantly lower ( P < 0.05). Exposure of organic dust is associated with compromised pulmonary functions and there is a need of formulation of safety guidelines.

FTIR Spectroscopy for Evaluation and Monitoring of Lipid Extraction Efficiency for Oleaginous Fungi.

PubMed

Forfang, Kristin; Zimmermann, Boris; Kosa, Gergely; Kohler, Achim; Shapaval, Volha

2017-01-01

To assess whether Fourier Transform Infrared (FTIR) spectroscopy could be used to evaluate and monitor lipid extraction processes, the extraction methods of Folch, Bligh and Lewis were used. Biomass of the oleaginous fungi Mucor circinelloides and Mortierella alpina were employed as lipid-rich material for the lipid extraction. The presence of lipids was determined by recording infrared spectra of all components in the lipid extraction procedure, such as the biomass before and after extraction, the water and extract phases. Infrared spectra revealed the incomplete extraction after all three extraction methods applied to M.circinelloides and it was shown that mechanical disruption using bead beating and HCl treatment were necessary to complete the extraction in this species. FTIR spectroscopy was used to identify components, such as polyphosphates, that may have negatively affected the extraction process and resulted in differences in extraction efficiency between M.circinelloides and M.alpina. Residual lipids could not be detected in the infrared spectra of M.alpina biomass after extraction using the Folch and Lewis methods, indicating their complete lipid extraction in this species. Bligh extraction underestimated the fatty acid content of both M.circinelloides and M.alpina biomass and an increase in the initial solvent-to-sample ratio (from 3:1 to 20:1) was needed to achieve complete extraction and a lipid-free IR spectrum. In accordance with previous studies, the gravimetric lipid yield was shown to overestimate the potential of the SCO producers and FAME quantification in GC-FID was found to be the best-suited method for lipid quantification. We conclude that FTIR spectroscopy can serve as a tool for evaluating the lipid extraction efficiency, in addition to identifying components that may affect lipid extraction processes.

An unusual case of anaphylaxis. Mold in pancake mix.

PubMed

Bennett, A T; Collins, K A

2001-09-01

Anaphylactic reactions involve contact with an antigen that evokes an immune reaction that is harmful. This type of reaction is a rapidly developing immunologic reaction termed a type I hypersensitivity reaction. The antigen complexes with an IgE antibody that is bound to mast cells and basophils in a previously sensitized individual. Upon re-exposure, vasoactive and spasmogenic substances are released that act on vessels and smooth muscle. The reaction can be local or systemic and may be fatal. The authors report the death of a 19-year-old white male who had a history of "multiple allergies," including pets, molds, and penicillin. One morning, he and his friends made pancakes with a packaged mix that had been opened and in the cabinet for approximately 2 years. The friends stopped eating the pancakes because they said that they tasted like "rubbing alcohol." The decedent continued to eat the pancakes and suddenly became short of breath. He was taken to a nearby clinic, where he became unresponsive and died. At autopsy, laryngeal edema and hyperinflated lungs with mucous plugging were identified. Microscopically, edema and numerous degranulating mast cells were identified in the larynx. The smaller airways contained mucus, and findings of chronic asthma were noted. Serum tryptase was elevated at 14.0 ng/ml. The pancake mix was analyzed and found to contain a total mold count of 700/g of mix as follows: Penicillium, Fusarium, Mucor, and Aspergillus. Witness statements indicate that the decedent ate two pancakes; thus he consumed an approximate mold count of 21,000. The decedent had a history of allergies to molds and penicillin, and thus was allergic to the molds in the pancake mix. The authors present this unusual case of anaphylaxis and a review of the literature.

Mycoalgae biofilm: development of a novel platform technology using algae and fungal cultures.

PubMed

Rajendran, Aravindan; Hu, Bo

2016-01-01

Microalgae is considered a promising source for biofuel and bioenergy production, bio-remediation and production of high-value bioactive compounds, but harvesting microalgae is a major bottleneck in the algae based processes. The objective of this research is to mimic the growth of natural lichen and develop a novel biofilm platform technology using filamentous fungi and microalgae to form a lichen type of biofilm "mycoalgae" in a supporting polymer matrix. The possibility of co-existence of Chlorella vulgaris with various fungal cultures was tested to identify the best strain combination for high algae harvest efficiency. The effect of different matrices for cell attachment and biofilm formation, cell surface characterization of mycoalgae biofilm, kinetics of the process with respect to the algae-fungi cell distribution and total biomass production was studied. Mycoalgae biofilm with algae attachment efficiency of 99.0 % and above was achieved in a polymer-cotton composite matrix with glucose concentration of 2 g/L in the growth medium and agitation intensity of 150 rpm at 27 °C. The total biomass in the co-culture with the selected strain combination (Mucor sp. and Chlorella sp.) was higher than the axenic cultures of fungi and algae at the conditions tested. The results show that algae can be grown with complete attachment to a bio-augmenting fungal surface and can be harvested readily as a biofilm for product extraction from biomass. Even though, interaction between heterotrophic fungi and phototrophic algae was investigated in solid media after prolonged contact in a report, this research is the first of its kind in developing an artificial lichen type biofilm called "mycoalgae" biofilm completely attached on a matrix in liquid cultures. The mycoalgae biofilm based processes, propounds the scope for exploring new avenues in the bio-production industry and bioremediation.

Impact of production systems on swine confinement buildings bioaerosols.

PubMed

Létourneau, Valérie; Nehmé, Benjamin; Mériaux, Anne; Massé, Daniel; Duchaine, Caroline

2010-02-01

Hog production has been substantially intensified in Eastern Canada. Hogs are now fattened in swine confinement buildings with controlled ventilation systems and high animal densities. Newly designed buildings are equipped with conventional manure handling and management systems, shallow or deep litter systems, or source separation systems to manage the large volumes of waste. However, the impacts of those alternative production systems on bioaerosol concentrations within the barns have never been evaluated. Bioaerosols were characterized in 18 modern swine confinement buildings, and the differences in bioaerosol composition in the three different production systems were evaluated. Total dust, endotoxins, culturable actinomycetes, fungi, and bacteria were collected with various apparatuses. The total DNA of the air samples was extracted, and quantitative polymerase chain reaction (PCR) was used to assess the total number of bacterial genomes, as a total (culturable and nonculturable) bacterial assessment. The measured total dust and endotoxin concentrations were not statistically different in the three studied production systems. In buildings with sawdust beds, actinomycetes and molds were found in higher concentrations than in the conventional barns. Aspergillus, Cladosporium, Penicillium, and Scopulariopsis species were identified in all the studied swine confinement buildings. A. flavus, A. terreus, and A. versicolor were abundantly present in the facilities with sawdust beds. Thermotolerant A. fumigatus and Mucor were usually found in all the buildings. The culturable bacteria concentrations were higher in the barns with litters than in the conventional buildings, while real-time PCR revealed nonstatistically different concentrations of total bacteria in all the studied swine confinement buildings. In terms of workers' respiratory health, barns equipped with a solid/liquid separation system may offer better air quality than conventional buildings or barns with

FTIR Spectroscopy for Evaluation and Monitoring of Lipid Extraction Efficiency for Oleaginous Fungi

PubMed Central

Zimmermann, Boris; Kosa, Gergely; Kohler, Achim; Shapaval, Volha

2017-01-01

To assess whether Fourier Transform Infrared (FTIR) spectroscopy could be used to evaluate and monitor lipid extraction processes, the extraction methods of Folch, Bligh and Lewis were used. Biomass of the oleaginous fungi Mucor circinelloides and Mortierella alpina were employed as lipid-rich material for the lipid extraction. The presence of lipids was determined by recording infrared spectra of all components in the lipid extraction procedure, such as the biomass before and after extraction, the water and extract phases. Infrared spectra revealed the incomplete extraction after all three extraction methods applied to M.circinelloides and it was shown that mechanical disruption using bead beating and HCl treatment were necessary to complete the extraction in this species. FTIR spectroscopy was used to identify components, such as polyphosphates, that may have negatively affected the extraction process and resulted in differences in extraction efficiency between M.circinelloides and M.alpina. Residual lipids could not be detected in the infrared spectra of M.alpina biomass after extraction using the Folch and Lewis methods, indicating their complete lipid extraction in this species. Bligh extraction underestimated the fatty acid content of both M.circinelloides and M.alpina biomass and an increase in the initial solvent-to-sample ratio (from 3:1 to 20:1) was needed to achieve complete extraction and a lipid-free IR spectrum. In accordance with previous studies, the gravimetric lipid yield was shown to overestimate the potential of the SCO producers and FAME quantification in GC-FID was found to be the best-suited method for lipid quantification. We conclude that FTIR spectroscopy can serve as a tool for evaluating the lipid extraction efficiency, in addition to identifying components that may affect lipid extraction processes. PMID:28118388

Increased virulence of Cunninghamella bertholletiae in experimental pulmonary mucormycosis: correlation with circulating molecular biomarkers, sporangiospore germination and hyphal metabolism.

PubMed

Petraitis, Vidmantas; Petraitiene, Ruta; Antachopoulos, Charalampos; Hughes, Johanna E; Cotton, Margaret P; Kasai, Miki; Harrington, Susan; Gamaletsou, Maria N; Bacher, John D; Kontoyiannis, Dimitrios P; Roilides, Emmanuel; Walsh, Thomas J

2013-01-01

Members of the order Mucorales are emerging invasive molds that cause infections in immunocompromised patients. However, little is known about the relation between different species of Mucorales and their virulence in invasive pulmonary mucormycosis. Based upon our earlier epidemiological studies, we hypothesized that Cunninghamella bertholletiae would demonstrate increased virulence. Therefore, we studied the relative virulence of C. bertholletiae (CB), Rhizopus oryzae (RO), R. microsporus (RM), and Mucor circinelloides (MC) in experimental invasive pulmonary mucormycosis in persistently neutropenic rabbits in relation to the fungi in vitro sporangiospore germination rate and hyphal metabolic activity. Rabbits infected with CB demonstrated (1) higher lung weights in comparison to RM (P ≤ 0.05), RO and MC (P ≤ 0.001), (2) pulmonary infarcts in comparison to RO and MC (P ≤ 0.001), (3) tissue fungal burden (CFU/g) vs. MC (P ≤ 0.001), and (4) the lowest survival of 0% (0/18), in comparison to 16% (3/18, P ≤ 0.01) of RM, 81% (21/26) of RO, and 83% (15/18) of MC-infected rabbits (P ≤ 0.001). Serum PCR concentration-time-curve showed the greatest amplitude for CB. Virulence correlated directly with sporangiospore germination rate at 4 h among species, i.e., CB (67-85%) > RM (14-56%) > RO (4-30%) > MC (0%), and hyphal metabolic activity, i.e., CB (1.22-1.51) > MC (0.54-0.64) = RM (0.38-0.41) = RO (0.37-0.59). C. bertholletiae was significantly more virulent in experimental invasive pulmonary mucormycosis than R. microsporus, R. oryzae, and M. circinelloides. In vivo virulence correlated with species-dependent differences of in vitro germination rate and hyphal metabolic activity.

Possible effects of glyphosate on Mucorales abundance in the rumen of dairy cows in Germany.

PubMed

Schrödl, Wieland; Krüger, Susanne; Konstantinova-Müller, Theodora; Shehata, Awad A; Rulff, Ramon; Krüger, Monika

2014-12-01

Glyphosate (N-phosphonomethyl glycine) is registered as a herbicide for many food and non-food crops, as well as non-crop areas where total vegetation control is desired. Glyphosate influences the soil mycobiota; however, the possible effect of glyphosate residues in animal feed (soybean, corn, etc.) on animal mycobiota is almost unknown. Accordingly, the present study was initiated to investigate the mycological characteristics of dairy cows in relationship to glyphosate concentrations in urine. A total of 258 dairy cows on 14 dairy farms in Germany were examined. Glyphosate was detected in urine using ELISA. The fungal profile was analyzed in rumen fluid samples using conventional microbiological culture techniques and differentiated by MALDI-TOF mass spectrometry. LPS-binding protein (LBP) and antibodies (IgG1, IgG2, IgA, and IgM) against fungi were determined in blood using ELISA. Different populations of Lichtheimia corymbifera, Lichtheimia ramosa, Mucor, and Rhizopus were detected. L. corymbifera and L. ramosa were significantly more abundant in animals containing high glyphosate (>40 ng/ml) concentrations in urine. There were no significant changes in IgG1 and IgG2 antibodies toward isolated fungi that were related to glyphosate concentration in urine; however, IgA antibodies against L. corymbifera and L. ramosa were significantly lower in the higher glyphosate groups. Moreover, a negative correlation between IgM antibodies against L. corymbifera, L. ramosa, and Rhizopus relative to glyphosate concentration in urine was observed. LBP also was significantly decreased in animals with higher concentrations of glyphosate in their urine. In conclusion, glyphosate appears to modulate the fungal community. The reduction of IgM antibodies and LBP indicates an influence on the innate immune system of animals.

Early diagnosis and monitoring of mucormycosis by detection of circulating DNA in serum: retrospective analysis of 44 cases collected through the French Surveillance Network of Invasive Fungal Infections (RESSIF).

PubMed

Millon, L; Herbrecht, R; Grenouillet, F; Morio, F; Alanio, A; Letscher-Bru, V; Cassaing, S; Chouaki, T; Kauffmann-Lacroix, C; Poirier, P; Toubas, D; Augereau, O; Rocchi, S; Garcia-Hermoso, D; Bretagne, S

2016-09-01

The main objective of this study was to assess the diagnostic performance of a set of three Mucorales quantitative PCR assays in a retrospective multicentre study. Mucormycosis cases were recorded thanks to the French prospective surveillance programme (RESSIF network). The day of sampling of the first histological or mycological positive specimen was defined as day 0 (D0). Detection of circulating DNA was performed on frozen serum samples collected from D-30 to D30, using quantitative PCR assays targeting Rhizomucor, Lichtheimia, Mucor/Rhizopus. Forty-four patients diagnosed with probable (n = 19) or proven (n = 25) mucormycosis were included. Thirty-six of the 44 patients (81%) had at least one PCR-positive serum. The first PCR-positive sample was observed 9 days (range 0-28 days) before diagnosis was made using mycological criteria and at least 2 days (range 0-24 days) before imaging. The identifications provided with the quantitative PCR assays were all concordant with culture and/or PCR-based identification of the causal species. Survival rate at D84 was significantly higher for patients with an initially positive PCR that became negative after treatment initiation than for patients whose PCR remained positive (48% and 4%, respectively; p <10 -6 ). The median time for complete negativity of PCR was 7 days (range 3-19 days) after initiation of l-AmB treatment. Despite some limitations due to the retrospective design of the study, we showed that Mucorales quantitative PCR could not only confirm the mucormycosis diagnosis when other mycological arguments were present but could also anticipate this diagnosis. Quantification of DNA loads may also be a useful adjunct to treatment monitoring. Copyright © 2015 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Chestnut green waste composting for sustainable forest management: Microbiota dynamics and impact on plant disease control.

PubMed

Ventorino, Valeria; Parillo, Rita; Testa, Antonino; Viscardi, Sharon; Espresso, Francesco; Pepe, Olimpia

2016-01-15

Making compost from chestnut lignocellulosic waste is a possible sustainable management strategy for forests that employs a high-quality renewable organic resource. Characterization of the microbiota involved in composting is essential to better understand the entire process as well as the properties of the final product. Therefore, this study investigated the microbial communities involved in the composting of chestnut residues obtained from tree cleaning and pruning. The culture-independent approach taken highlighted the fact that the microbiota varied only slightly during the process, with the exception of those of the starting substrate and mature compost. The statistical analysis indicated that most of the bacterial and fungal species in the chestnut compost persisted during composting. The dominant microbial population detected during the process belonged to genera known to degrade recalcitrant lignocellulosic materials. Specifically, we identified fungal genera, such as Penicillium, Fusarium, Cladosporium, Aspergillus and Mucor, and prokaryotic species affiliated with Bacilli, Actinobacteria, Flavobacteria and γ-Proteobacteria. The suppressive properties of compost supplements for the biocontrol of Sclerotinia minor and Rhizoctonia solani were also investigated. Compared to pure substrate, the addition of compost to the peat-based growth substrates resulted in a significant reduction of disease in tomato plants of up to 70 % or 51 % in the presence of Sclerotinia minor or Rhizoctonia solani, respectively. The obtained results were related to the presence of putative bio-control agents and plant growth-promoting rhizobacteria belonging to the genera Azotobacter, Pseudomonas, Stenotrophomonas, Bacillus, Flavobacterium, Streptomyces and Actinomyces in the chestnut compost. The composting of chestnut waste may represent a sustainable agricultural practice for disposing of lignocellulosic waste by transforming it into green waste compost that can be used to

Acute invasive fungal rhinosinusitis: our experience with 18 cases.

PubMed

Bakhshaee, Mehdi; Bojdi, Amin; Allahyari, Abolghasem; Majidi, Mohammad Reza; Tavakol, Sherwin; Najafzadeh, Mohammad Javad; Asghari, Masoud

2016-12-01

Acute invasive fungal rhinosinusitis (AIFRS) is a rapidly progressive life threatening infection that is seen most commonly among immunocompromised patients. We present a case series of 18 patients clinically and histopathologically diagnosed with AIFRS with a mean follow-up of 9.11 ± 2.51 months (range 6-17). Demographic data, apparent symptoms and signs, underlying disorders, and outcomes are discussed. The mean age was 39.56 ± 20.66 years (range 2-75). The most common underlying diseases were diabetes mellitus (50 %) and leukemia (44.44 %). Mucosal biopsy confirmed fungal invasion of the nasal mucosa in all cases. The main fungi were Rhizopus oryzae (55.56 %), Absidia mucor (16.67 %), and Aspergillus fumigatus (27.78 %). Headache and facial pain (77.8 %), facial paresthesia (55.6 %), and ophthalmoplegia (33.3 %) were the most common symptoms and signs. Computed tomography and endoscopic findings showed various stages of sinonasal (100 %), pterygopalatine fossa (55.56 %), orbital (44.45 %), and cerebral (5.56 %) involvement. All patients underwent serial surgical debridement (3.78 ± 1.80 times; range 2-8) simultaneously with systemic antifungal therapy and proper management of the underlying disease. The most extreme case with brain involvement survived and recovered with no evidence of recurrent disease following treatment. All patients were considered cured after two endoscopic negative histopathologic evaluations. Three patients (16.67 %) died, one from uncontrolled leukemia and two due to renal failure. AIFRS is a potentially fatal condition, however, early diagnosis and management of the underlying disease accompanied with systemic antifungal and aggressive serial surgical intervention appears to be effective in reducing mortality in most patients.

Mould routine identification in the clinical laboratory by matrix-assisted laser desorption ionization time-of-flight mass spectrometry.

PubMed

Cassagne, Carole; Ranque, Stéphane; Normand, Anne-Cécile; Fourquet, Patrick; Thiebault, Sandrine; Planard, Chantal; Hendrickx, Marijke; Piarroux, Renaud

2011-01-01

MALDI-TOF MS recently emerged as a valuable identification tool for bacteria and yeasts and revolutionized the daily clinical laboratory routine. But it has not been established for routine mould identification. This study aimed to validate a standardized procedure for MALDI-TOF MS-based mould identification in clinical laboratory. First, pre-extraction and extraction procedures were optimized. With this standardized procedure, a 143 mould strains reference spectra library was built. Then, the mould isolates cultured from sequential clinical samples were prospectively subjected to this MALDI-TOF MS based-identification assay. MALDI-TOF MS-based identification was considered correct if it was concordant with the phenotypic identification; otherwise, the gold standard was DNA sequence comparison-based identification. The optimized procedure comprised a culture on sabouraud-gentamicin-chloramphenicol agar followed by a chemical extraction of the fungal colonies with formic acid and acetonitril. The identification was done using a reference database built with references from at least four culture replicates. For five months, 197 clinical isolates were analyzed; 20 were excluded because they were not identified at the species level. MALDI-TOF MS-based approach correctly identified 87% (154/177) of the isolates analyzed in a routine clinical laboratory activity. It failed in 12% (21/177), whose species were not represented in the reference library. MALDI-TOF MS-based identification was correct in 154 out of the remaining 156 isolates. One Beauveria bassiana was not identified and one Rhizopus oryzae was misidentified as Mucor circinelloides. This work's seminal finding is that a standardized procedure can also be used for MALDI-TOF MS-based identification of a wide array of clinically relevant mould species. It thus makes it possible to identify moulds in the routine clinical laboratory setting and opens new avenues for the development of an integrated MALDI-TOF MS

A novel library-independent approach based on high-throughput cultivation in Bioscreen and fingerprinting by FTIR spectroscopy for microbial source tracking in food industry.

PubMed

Shapaval, V; Møretrø, T; Wold Åsli, A; Suso, H P; Schmitt, J; Lillehaug, D; Kohler, A

2017-05-01

Microbiological source tracking (MST) for food industry is a rapid growing area of research and technology development. In this paper, a new library-independent approach for MST is presented. It is based on a high-throughput liquid microcultivation and FTIR spectroscopy. In this approach, FTIR spectra obtained from micro-organisms isolated along the production line and a product are compared to each other. We tested and evaluated the new source tracking approach by simulating a source tracking situation. In this simulation study, a selection of 20 spoilage mould strains from a total of six genera (Alternaria, Aspergillus, Mucor, Paecilomyces, Peyronellaea and Phoma) was used. The simulation of the source tracking situation showed that 80-100% of the sources could be correctly identified with respect to genus/species level. When performing source tracking simulations, the FTIR identification diverged for Phoma glomerata strain in the reference collection. When reidentifying the strain by sequencing, it turned out that the strain was a Peyronellaea arachidicola. The obtained results demonstrated that the proposed approach is a versatile tool for identifying sources of microbial contamination. Thus, it has a high potential for routine control in the food industry due to low costs and analysis time. The source tracking of fungal contamination in the food industry is an important aspect of food safety. Currently, all available methods are time consuming and require the use of a reference library that may limit the accuracy of the identification. In this study, we report for the first time, a library-independent FTIR spectroscopic approach for MST of fungal contamination along the food production line. It combines high-throughput microcultivation and FTIR spectroscopy and is specific on the genus and species level. Therefore, such an approach possesses great importance for food safety control in food industry. © 2016 The Society for Applied Microbiology.

Antimicrobial, antiplasmodial, haemolytic and antioxidant activities of crude extracts from three selected Togolese medicinal plants.

PubMed

Karou, Simplice D; Tchacondo, Tchadjobo; Ouattara, Lassina; Anani, Kokou; Savadogo, Aly; Agbonon, Amegnona; Attaia, Mossaclok Ben; de Souza, Comlan; Sakly, Mohsen; Simpore, Jacques

2011-10-01

To investigate the antioxidant, antimicrobial, antiplasmodial, acute toxicity and haemolytic activities of methanolic extracts of three plants. Phytochemical analysis to determine the phenolic contents was also carried out. The 2,2-diphenyl-1-picryl-hydrazyl (DPPH) free radical scavenging, NCCLS broth microdilution and Plasmodium Lactate Dehydrogenase (pLDH) assays were used to determine antioxidant, antimicrobial and antiplasmodial activities, respectively. Haemolysis assay was conducted on A(+) human red blood cells and acute toxicity on male Swiss albino mice. Phenolics were quantitatively determined using spectrophotometric methods. The DPPH assay yielded interesting antioxidant activities of methanolic extract of Parinari curatellifolia (P. curatellifolia) and Entada africana (E. africana) (IC(50) were 0.20±0.01 μg/mL and 0.47±0.01 μg/mL, respectively). This activity was highly correlated with phenolic contents of extracts. The antimicrobial tests displayed minimal inhibitory concentrations (MICs) values ranging from 0.90 to 1.80 mg/mL for Serratia marcescens (S. marcescens) the most susceptible bacterial strain. MIC value was 1.20 mg/mL for susceptible fungal strains including Mucor rouxi (M. rouxi), Fusarium oxyporum (F. oxyporum) and Rhizopus nigricans (R. nigricans). pLDH assay showed moderate antiplasmodial activity of Balanites aegyptiaca (B. aegyptiaca) (IC(50) = 24.56±3.45 μg/mL), however this extract was highly haemolytic and toxic in mice (LD(50) = 625±128 mg/kg). Our results support in part the use of the selected plants in the treatment of microbial infections. In addition the plant showed interesting antioxidant activity that could be useful in the management of oxidative stress. Copyright © 2011 Hainan Medical College. Published by Elsevier B.V. All rights reserved.

Effect of naturally mouldy wheat or fungi administration on metallothioneins level in brain tissues of rats.

PubMed

Vasatkova, Anna; Krizova, Sarka; Krystofova, Olga; Adam, Vojtech; Zeman, Ladislav; Beklova, Miroslava; Kizek, Rene

2009-01-01

The aim of this study is to determine level of metallothioneins (MTs) in brain tissues of rats administered by feed mixtures with different content of mouldy wheat or fungi. Selected male laboratory rats of Wistar albino at age of 28 days were used in our experiments. The rats were administered by feed mixtures with different content of vitamins, naturally mouldy wheat or fungi for 28 days. At the very end of the experiment, the animals were put to death and brains were sampled. MT level was determined by differential pulse voltammetry Brdicka reaction. We found that MTs' level in brain tissues from rats administered by standard feed mixtures was significantly higher compared to the level of MTs in rats supplemented by vitamins. Further we studied the effect of supplementation of naturally mouldy wheat on MTs level in rats. In mouldy wheat we detected the presence of following fungi species: Mucor spp., Absidia spp., Penicillium spp., Aspergillus spp. and Fusarium spp. Moreover we also identified and quantified following mycotoxins - deoxynivalenol, zearalenone, T2-toxin and aflatoxins. Level of MTs determined in rats treated with 33 or 66% of mouldy wheat was significantly lower compared to control ones. On the other hand rats treated with 100% of mouldy wheat had less MTs but not significantly. Supplementation of vitamins to rats fed by mouldy wheat had adverse effect on MTs level compared to rats with no other supplementation by vitamins. Moreover vitamins supplementation has no effect on MTs level in brain tissues of rats treated or non-treated with Ganoderma lucidum L. Both mycotoxins and vitamins have considerable effect on level of MTs in brain tissues. It can be assumed that the administered substances markedly influence redox metabolism, which could negatively influence numerous biochemical pathways including those closely related with MTs.

A combined effort to avoid strongyle infection in horses in an oceanic climate region: rotational grazing and parasiticidal fungi.

PubMed

Hernández, José Ángel; Sánchez-Andrade, Rita; Cazapal-Monteiro, Cristiana Filipa; Arroyo, Fabián Leonardo; Sanchís, Jaime Manuel; Paz-Silva, Adolfo; Arias, María Sol

2018-04-12

An approach to preventing strongyle infection in horses was tested, comprising rotational pasturing and the administration of spores of two parasiticidal fungi, Mucor circinelloides and Duddingtonia flagrans. Twenty-two adult Spanish Sport Horses were dewormed with ivermectin (1 mg pour-on/kg body weight) and then randomly divided into three groups. G-1 was maintained with continuous grazing, and G-2 and G-3 were kept on a four-paddock rotation system. Commercial pelleted feed (2.5 kg/horse) was supplied to G-1 and G-2 twice a week; horses in G-3 received pellets containing 2 × 10 6 spores/kg of each fungus. Fecal samples were analyzed by the flotation method to estimate the reduction in the fecal egg counts (FECR), the percentage of horses shedding eggs (PHR), and the egg reappearance period (ERP). Third-stage larvae were identified in fecal pats as Cyathostomum (sensu lato) types A, C and D, Gyalocephalus capitatus, Triodontophorus serratus, Poteriosthomum spp., Strongylus vulgaris and S. edentatus. Two weeks after treatment, the FECR values were 100% in G-1, 96% in G-2 and 99% in G-3; the PHR values were 100% in G-1, 75% in G-2 and 88% in G-3. A strongyle ERP of 6 weeks was observed in G-1, ERP of 10 weeks was observed in G-2, and ERP of 16 weeks was observed in G-3. The counts of eggs per gram of feces (EPG) were > 300 EPG in G-1 and G-2 but remained below 250 EPG in G-3 throughout the observation period of 12 months. These results suggest that horse strongyle infection could be decreased by combining rotational pasturing with feeding pellets containing the spores of parasiticidal fungi.

Seasonal study of the fungal biota of the fur of dogs.

PubMed

Cabañes, F J; Abarca, M L; Bragulat, M R; Castellá, G

1996-01-01

During a one year period, 944 dogs from the Municipal kennel of Barcelona were examined to detect animals with suspected dermatophytosis. Only a few animals (1.8%) presented skin lesions but none of them had dermatophytosis. A representative number of dogs without visible skin lesions (n = 172), selected at random, were used to carry out a seasonal study of the mycobiota of their fur. Fifteen isolates belonging to the genera Microsporum and Trichophyton were isolated from 14 of the 172 (8.1%) dogs without lesions. The identity of these fungi was Microsporum gypseum (6/15), Trichophyton terrestre (4/15), M. canis (2/15), M. cookei (2/15) and Trichophyton ajelloi (1/15) (one strain each of M. gypseum and T. ajelloi were isolated from one dog). Species of Penicillium (% prevalence = 89.5%), Alternaria (86.6%), Cladosporium (84.9%), Aspergillus (77.3%), Scopulariopsis (65.7%) and Chrysosporium (64.5%) were the most prevalent. No significant differences in the fungal biota were observed with respect to age, gender, hair length or between mixed and pure breed dogs. A large number of isolates, including species belonging to the genera Beauveria, Chrysosporium, Malbranchea and Scopulariopsis, that macroscopically and/or microscopically resemble dermatophytes and may be mistaken for them, produced a red color change in Dermatophyte Test Medium. No significant seasonal difference was detected among the isolates belonging to the most frequently encountered genera, with the exception of Scopulariopsis (higher in summer and autumn) and Chrysosporium (higher in summer). Species from other genera, with lower occurrence also presented significant differences in their seasonal distribution. Arthrinium, Aureobasidium, Chaetomium and Phoma spp. presented maximum prevalence peaks in spring, Fusarium, Paecilomyces, Phoma and Rhizopus spp. in summer and Geotrichum and Mucor spp. in autumn. The Microsporum and Trichophyton species were more frequently isolated in summer.

Microflora inside closed modules with plant growth facility

NASA Astrophysics Data System (ADS)

Zyablova, Natalya V.; Berkovich, Yuliy A.; Shanturin, Nikolai; Deshevaya, Elena; Smolyanina, Svetlana O.

Currently, plant growth facility (PGF) is included in the LSS in many scenarios of Martian expedition. A number of investigators assume growing of crops can accelerate microflora re-production in closed ecological system. To estimate experimentally the change of density of microbiological community in the isolated module, Chinese cabbage Brassica hinensis L., cv. Vesnyanka, has been grown in the closed climatic chambers in volume 0.07 m3, 3 m3 and 250 m3 under continuous illumination in the range of values of temperature and relative humidity of air 23 -270 and 30 -60%, respectively. There were no differences in growth and develop-ment of plants grown during 30 days on the test-beds in the laboratory room (control) and in the closed chamber by 0.07 m3 volume (test). The microbiological analysis of root zone has revealed the presence of exclusively saprophytic species -the typical representatives of the soil microbiota. Then the plants were growing during 45 days in the prototype of the conveyor space PGF "Phytocycle LED" placed inside the chamber of 3 m3 volume. Every 3 days 50 -60 cm3 of liquid imitator of air condensate (IAC) from inhabited module had been injected to the chamber to simulate air pollution. The content of colony-forming units of the micromycetes in the air of the chamber, on the inner surfaces of the climate chamber, internal and external surfaces of the PGF and the leaves did not exceed the permissible values. When the PGF has been installed during 14 days inside the inhabited module with volume of 250 m3, the representatives of saprophytic and conditioned-pathogenic species of micromycetes (Trichethe-cium rozeum, Trichoderma sp., Fuzarrium sp., Mucor sp., Penicillium sp.) have been found out exclusively on the open surfaces of artificial soil and water-saturated porous passage. The obtained data shows that PGF inside closed modules can assure microbiological safety when all wet surfaces are isolated from the gas environment.

Fungal dissemination by housefly (Musca domestica L.) and contamination of food commodities in rural areas of South Africa.

PubMed

Phoku, J Z; Barnard, T G; Potgieter, N; Dutton, M F

2016-01-18

Several insects that act as vectors, including houseflies (Musca domestica L.), are often considered to be an important source of fungal contamination in human foods. Houseflies are also involved in the transmission of bacterial pathogens that may pose a serious hazard to human health. Thus, the rural population of South Africa, as typified by that in the Gauteng Province investigated in this study, is at high risk from fungal exposure disseminated by houseflies and it is therefore important to assess the role of flies in contaminating various food commodities. Eighty four samples of houseflies (captured from households and pit toilets) were studied for their potential to carry fungal spores into food commodities. The fungi occurring in samples of raw maize (15) and porridge (19) were also assessed. Fungal isolates were identified based on morphological characteristics by conventional identification methods. Fifteen genera of fungi were isolated and identified, of which Aspergillus, Fusarium, Penicillium, Cladosporium, Moniliella and Mucor were the most prevalent in all three sample types analysed. The incidence rates of fungal contamination per total fungal count isolated in houseflies, maize and porridge were recorded with mean fungal load of 2×10(8) CFU/ml, 1×10(7)CFU/g and 2×10(7)CFU/g respectively. Additionally, A. flavus, A. parasiticus, F. verticillioides, F. proliferatum, P. verrucosum, P. aurantiogriseum and M. suaveolens were the most frequent fungal isolates in houseflies with incidence rate of 34%, 11%, 27%, 21%, 22%, 17% and 32% respectively. F. verticillioides, A. flavus, A. niger and P. oslonii were the most prevalent species contaminating porridge and maize with incidence rate of 23%, 32%, 16% and 28% in maize samples, while incidence rates of 59%, 15% and 29% were recorded in porridge samples with the exception of F. verticillioides. The prevalence of these genera of fungi may pose serious health risks. Copyright © 2015 Elsevier B.V. All rights

Bioremediation of 60Co from simulated spent decontamination solutions.

PubMed

Rashmi, K; Sowjanya, T Naga; Mohan, P Maruthi; Balaji, V; Venkateswaran, G

2004-07-26

Bioremediation of 60Co from simulated spent decontamination solutions by utilizing different biomass of (Neurospora crassa, Trichoderma viridae, Mucor recemosus, Rhizopus chinensis, Penicillium citrinum, Aspergillus niger and, Aspergillus flavus) fungi is reported. Various fungal species were screened to evaluate their potential for removing cobalt from very low concentrations (0.03-0.16 microM) in presence of a high background of iron (9.33 mM) and nickel (0.93 mM) complexed with EDTA (10.3 mM). The different fungal isolates employed in this study showed a pickup of cobalt in the range 8-500 ng/g of dry biomass. The [Fe]/[Co] and [Ni]/[Co] ratios in the solutions before and after exposure to the fungi were also determined. At micromolar level the cobalt pickup by many fungi especially the mutants of N. crassa is seen to be proportional to the initial cobalt concentration taken in the solution. However, R. chinensis exhibits a low but iron concentration dependent cobalt pickup. Prior saturating the fungi with excess of iron during their growth showed the presence of selective cobalt pickup sites. The existence of cobalt specific sorption sites is shown by a model experiment with R. chinensis wherein at a constant cobalt concentration (0.034 microM) and varying iron concentrations so as to yield [Fe/Co]initial ratios in solution of 10, 100, 1000 and 287000 have all yielded a definite Co pickup capacity in the range 8-47 ng/g. The presence of Cr(III)EDTA (3 mM) in solution along with complexed Fe and Ni has not influenced the cobalt removal. The significant feature of this study is that even when cobalt is present in trace level (sub-micromolar) in a matrix of high concentration (millimolar levels) of iron, nickel and chromium, a situation typically encountered in spent decontamination solutions arising from stainless steel based primary systems of nuclear reactors, a number of fungi studied in this work showed a good sensitivity for cobalt pickup. Copyright 2004

Health Threats from Contamination of Spices Commercialized in Romania: Risks of Fungal and Bacterial Infections.

PubMed

Man, Adrian; Mare, Anca; Toma, Felicia; Curticăpean, Augustin; Santacroce, Luigi

2016-01-01

The study of fungal contamination in food and mycotoxicoses is a priority today, both internationally and nationally. The purpose of this study is to have a general view over the quality of the most common spices that are sold in Romanian markets, by assessing the degree of fungal, bacterial and mycotoxin contamination in pepper and chili powders. We tested four types of spices: white pepper, black pepper, sweet and hot chili powders from 12 different distributing companies, summing a total of 35 sample types. The fungal and bacterial load was assessed by Standard Plate Count, while the mycotoxin content by High-performance liquid chromatography. Environmental conditions (humidity, pH) and the selling price for each product were also followed. Fungi were observed in 72.7% of black pepper samples, 33.3% in white pepper, 30% in sweet chili and 25% in hot chili products. The most common isolated fungus was Aspergillus spp., while Rhizopus, Mucor, Fusarium, Penicillium, Absidia species were found, in smaller percentage. Four producers (44.4%) presented fungal contamination of over 10^3 CFU/g and two producers (22.2%) presented no fungal contamination in their products. Bacterial contamination was found in 85.7% of the tested products, consisting mostly in Bacillus spp. Aflatoxin B1 was present in all the tested products, mostly in black pepper (mean value 126.3 ng/g); Ochratoxin A was present in sweet chili (mean value 328 ng/g) and Zearalenone in hot chili (mean value 604 ng/g) and sweet chili (mean value 382 ng/g). All spices presented either fungal contamination, mycotoxin contamination, or both. The high humidity and the high pH of spices represent favorable conditions for fungal growth. The selling price was partly related to the physic-chemical conditions and microbiological quality of the spices. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.org.

Cork Taint of Wines: Role of the Filamentous Fungi Isolated from Cork in the Formation of 2,4,6-Trichloroanisole by O Methylation of 2,4,6-Trichlorophenol

PubMed Central

Álvarez-Rodríguez, María Luisa; López-Ocaña, Laura; López-Coronado, José Miguel; Rodríguez, Enrique; Martínez, María Jesús; Larriba, Germán; Coque, Juan-José R.

2002-01-01

Cork taint is a musty or moldy off-odor in wine mainly caused by 2,4,6-trichloroanisole (2,4,6-TCA). We examined the role of 14 fungal strains isolated from cork samples in the production of 2,4,6-TCA by O methylation of 2,4,6-trichlorophenol (2,4,6-TCP). The fungal strains isolated belong to the genera Penicillium (four isolates); Trichoderma (two isolates); and Acremonium, Chrysonilia, Cladosporium, Fusarium, Mortierella, Mucor, Paecilomyces, and Verticillium (one isolate each). Eleven of these strains could produce 2,4,6-TCA when they were grown directly on cork in the presence of 2,4,6-TCP. The highest levels of bioconversion were carried out by the Trichoderma and Fusarium strains. One strain of Trichoderma longibrachiatum could also efficiently produce 2,4,6-TCA in liquid medium. However, no detectable levels of 2,4,6-TCA production by this strain could be detected on cork when putative precursors other than 2,4,6-TCP, including several anisoles, dichlorophenols, trichlorophenols, or other highly chlorinated compounds, were tested. Time course expression studies with liquid cultures showed that the formation of 2,4,6-TCA was not affected by a high concentration of glucose (2% or 111 mM) or by ammonium salts at concentrations up to 60 mM. In T. longibrachiatum the O methylation of 2,4,6-TCP was catalyzed by a mycelium-associated S-adenosyl-l-methionine (SAM)-dependent methyltransferase that was strongly induced by 2,4,6-TCP. The reaction was inhibited by S-adenosyl-l-homocysteine, an inhibitor of SAM-dependent methylation, suggesting that SAM is the natural methyl donor. These findings increase our understanding of the mechanism underlying the origin of 2,4,6-TCA on cork, which is poorly understood despite its great economic importance for the wine industry, and they could also help us improve our knowledge about the biodegradation and detoxification processes associated with chlorinated phenols. PMID:12450804

Airway toxicity of house dust and its fungal composition.

PubMed

Piecková, Elena; Wilkins, Ken

2004-01-01

House dust is an important source of different toxic metabolites as well as allergens, including those of fungal origin, in the indoor environment. A bio-assay employing 1-day-old chick tracheas was used to characterize airway effects of 2-butanone and dimethylsulphoxide (Me2SO) extracts of 23 dust samples collected from water damaged (13) and control (10) Danish schools. Direct microscopical analysis of samples, followed by cultivation on dichloran 18 % glycerol agar at 25 degrees C for 10 days to establish their mycoflora, was performed. The in vitro ciliostatic potential of the chloroform-extractable endo- and exometabolites of 41 representative fungal isolates was determined. Nine dust extracts in 2-butanone (2 from damp rooms) or 10 (6) in Me(2)SO showed some ciliostatic activity in the 3-days' experiment. Fungal composition of dust from buildings with leakage was almost identical with that from undamaged houses, as well as the fungal colony counts from the damp schools and the control samples. Aspergillus spp. were prevalent in the samples - 31 or 40 % of all fungi, followed by Penicillium spp. and Cladosporium cladosporioides. Alternaria spp., Chaetomium spp., Mucor spp., Mycelia sterilia, Paecilomyces variotii, Rhizopus sp., Ulocladium sp. and yeasts were each isolated in less than 8 % of the fungal content. No Aspergillus flavus isolate (8 in total) was aflatoxigenic,em>in vitro. Alternaria spp., Aspergillus spp., Botrytis cinerea, Penicillium spp., C. cladosporioides, Chaetomium spp. and Ulocladium sp.; in total, 88 % of all fungi tested, produced ciliostatically active metabolites. These toxigenic strains were also present in 4 dust samples from controls and 5 dust samples from water damaged buildings. Extracts of these dust samples were also toxic in bioassay. There were bio-detectable concentrations (10-20 microg of extracts/ml of the organ culture medium) of toxic compounds in house dust. Contribution of fungal metabolites to its toxic effect should

In vitro antioxidant potential of selected aphrodisiac medicinal plants.

PubMed

Riaz, M; Shahid, M; Jamil, A; Saqib, M

2017-01-01

The present study aimed to evaluate the antioxidant activity of six selected aphrodisiac medicinal plants. Useful parts of the selected medicinal plants were collected and extracted in methanolic solvent. The antioxidant activity of selected plant extract was determined through different antioxidant assays, namely DPPH radical scavenging assay and ferric reducing antioxidant assay. Moreover, antioxidant compounds, like total phenolics and total flavonoids contents, were also determined. Results showed that Mucuna pruriens seed extract displayed high contents of phenolic compounds with total phenolic content of 683.15±4.28 mg GAE/g dry plant material while the least phenolic content was observed in Asparagus racemosus (195.5±3.02 mg GAE/g dry plant material). Highest total flavonoids content was found in Anacyclus pyrethrum roots (156.58±4.01 μg CE/g) and the least content was found in Asparagus racemosus roots. Among the studied plant extracts, the highest radical scavenging activity was shown by Mucuna pruriens seed extract (82.05±0.55%) and the least percent scavenging activity was observed in Tribulus terrestris extract (36.40±2.01%). Vitamin C was used as positive control for antioxidant assays showing 93.54±0.9% radical scavenging activity. The plant extract also exhibited a strong reducing potential against free radicals. Therefore, the present study concluded that all the studied medicinal plants possess varying concentrations of secondary active metabolites responsible for the antioxidant properties of the tested plant extracts.

Selenium hyperaccumulation by Astragalus (Fabaceae) does not inhibit root nodule symbiosis.

PubMed

Alford, Elan R; Pilon-Smits, Elizabeth A H; Fakra, Sirine C; Paschke, Mark W

2012-12-01

A survey of the root-nodule symbiosis in Astragalus and its interaction with selenium (Se) has not been conducted before. Such studies can provide insight into how edaphic conditions modify symbiotic interactions and influence partner coevolution. In this paper plant-organ Se concentration ([Se]) was investigated to assess potential Se exposure to endophytes. • Selenium distribution and molecular speciation of root nodules from Se-hyperaccumulators Astragalus bisulcatus, A. praelongus, and A. racemosus was determined by Se K-edge x-ray absorption spectroscopy. A series of greenhouse experiments were conducted to characterize the response of root-nodule symbiosis in Se-hyperaccumulators and nonhyperaccumulators. • Nodules in three Se-hyperaccumulators (Astragalus crotalariae, A. praelongus, and A. preussii) are reported for the first time. Leaves, flowers, and fruits from Se-hyperaccumulators were routinely above the hyperaccumulator threshold (1,000 µg Se g(-1) DW), but root samples rarely contained that amount, and nodules never exceeded 110 µg Se g(-1) DW. Nodules from A. bisulcatus, A. praelongus, and A. racemosus had Se throughout, with a majority stored in C-Se-C form. Finally, an evaluation of nodulation in Se-hyperaccumulators and nonhyperaccumulators indicated that there was no nodulation inhibition because of plant Se tolerance. Rather, we found that in Se-hyperaccumulators higher levels of Se treatment (up to 100 µM Se) corresponded with higher nodule counts, indicating a potential role for dinitrogen fixation in Se-hyperaccumulation. The effect was not found in nonhyperaccumulators. • As the evolution of Se hyperaccumulation in Astragalus developed, root-nodule symbiosis may have played an integral role.

Mineral-microorganism interactions in Acid Mine Drainage environments: preliminary results

NASA Astrophysics Data System (ADS)

Carbone, Cristina; Zotti, Mirca; Pozzolini, Marina; Giovine, Marco; Di Piazza, Simone; Mariotti, Mauro; Lucchetti, Gabriella

2014-05-01

Minerals play a key role in controlling the mobility and distribution of metals and metalloids of environmental concern in supergenic environments. These are involved in a variety of processes, spanning the alteration of primary minerals to the formation of secondary authigenic phases and can represent a source or a trap for Potentially Ecotoxic Elements (PTEs). Soil, sediments, and waters heavily polluted with PTEs through AMD processes are a reservoir of a unusual bacteria and fungi well adapted to these toxic environments. Classical studies of biotic weathering have mainly focused on water-mineral interaction and on the ability of microorganism to influence the soil solution chemical composition. In this work, we analyzed two different representative ochreous and greenish-blue AMD colloidal precipitates in order to i) characterize the biota population present in these colloidal minerals and ii) verify the bioaccumulation of PTEs into the fungi and the potential impact of bacteria in the geochemistry of the system. The samples are composed by nanocrystalline goethite which contains high amounts of Fe, Cu, Zn, Pb, and Ni and woodwardite that is characterized by Cu, Zn, Ni, Y, and Ce. These precipitates were examined in order to evaluate the presence of fungal strains and to extract bacteria DNA. The preliminary results of fungi characterization show an interesting and selected mycobiota able to survive under unfavourable environmental conditions. A significant number of fungal strains was isolated in pure culture. Most of them belong to the genus Mucor and Penicillium. It is worth noting the presence of Trametes versicolor, a macrofungal lignicolous species already known for heavy metal biosorption capability from aqueous solution (Gülay et al 2003). The same colloidal precipitates have been processed to extract bacteria DNA, using a specific procedure developed for DNA extraction from sediments. The results gave a good yield of nucleic acids and the positive PCR

Fungi in housefly (Musca domestica L.) as a disease risk indicator-A case study in South Africa.

PubMed

Phoku, J Z; Barnard, T G; Potgieter, N; Dutton, M F

2014-12-01

Houseflies are the commonest insects which have increasingly overcrowded human dwellings, particularly in rural areas and constitute a health hazard. In the environment they move back and forth by feeding and breeding on food commodities and filth. This may lead to the spread of diseases and also mycotoxin-producing fungi. Thus frequent exposure to the activity of such houseflies will have an impact on the welfare of humans. The study investigated the natural occurrence of fungal contamination in housefly samples captured from different households and pit toilets from a rural community in South Africa. Fungal contamination data were based on the prevalence, contamination level and morphological characteristics of the different identified species. A total of 497 fungal isolates of 15 genera including Aspergillus (37%), Fusarium (17%), Penicillium (21%), Alternaria (1.4%), Chrysosporium (2%), Cladosporium (0.2%), Curvularia (0.4%), Epicoccum (1%), Eupenicillium (1%), Moniliella (9%), Mucor (2%), Nigrospora (1%), Rhizopus (2%), Scopulariopsis (2%) and Yeasts (3%) were identified from the external surfaces of both female and male houseflies. The range of fungal contamination per total fungal count isolated from female and male houseflies were recorded with mean fungal load of 4.1×10(6), 8.4×10(6), 4.4×10(6), 3.3×10(5), 9.8×10(6), 2.2×10(4), 5.6×10(4), 2.9×10(6), 5.2×10(6), 4.7×10(6), 4.5×10(7), 4.6×10(6), 2.3×10(6), 4.9×10(7) and 6.4×10(6)CFU/ml, respectively. However, the range from The most dominant fungal isolates of the female housefly samples were Aspergillus flavus, Fusarium verticillioides, Penicillium verrucosum and Moniliella suaveolens, while A. flavus, Aspergillus parasiticus, F. verticillioides, Fusarium proliferatum and Penicillium aurantiogriseum were most prevalent in male samples. The study proves that housefly is a vector for fungal spores. Therefore, it is crucial to implement housefly-control measures to curb the spread of diseases

Feeding horses with industrially manufactured pellets with fungal spores to promote nematode integrated control.

PubMed

Hernández, José Ángel; Arroyo, Fabián Leonardo; Suárez, José; Cazapal-Monteiro, Cristiana Filipa; Romasanta, Ángel; López-Arellano, María Eugenia; Pedreira, José; de Carvalho, Luis Manuel Madeira; Sánchez-Andrade, Rita; Arias, María Sol; de Gives, Pedro Mendoza; Paz-Silva, Adolfo

2016-10-15

The usefulness of pellets industrially manufactured with spores of parasiticide fungi as a contribution to integrated nematode control was assessed in grazing horses throughout sixteen months. Two groups of 7 Pura Raza Galega autochthonous horses (G-T and G-P) were dewormed pour-on (1mg Ivermectin/kg bw) at the beginning of the trial, and other group (G-C) remained untreated. The G-P was provided daily with commercial pellets to which was added a mixture of fungal spores during the industrial manufacturing (2×10 6 spores of Mucor circinelloides and same dose of Duddingtonia flagrans/kg), and G-T and G-C received pellets without spores. The efficacy of the parasiticidal strategy was assessed by estimating the reduction in the faecal egg counts (FECR) and in the number of horses shedding eggs in the faeces (PHR), and also the egg reappearance periods (ERP). Blood analyses were performed to identify the changes in the red and white cell patterns. To ascertain if horses developed an IgG humoral response against the fungi, antigenic products collected from M. circinelloides and D. flagrans were exposed to the horse sera by using an ELISA. The faecal elimination of eggs of Parascaris equorum and strongyles ceased 2 weeks after treatment in G-T and G-P, thus the values of FECR and PHR were 100%. No P. equorum-eggs were detected later, and the strongyle egg reappearance period was 28 weeks in G-P, and 8 weeks in G-T. Strongyle egg-output values remained lower than 300 eggs per gram of faeces in the G-P, whereas numbers between 330 and 772 in G-C and G-T were recorded. Normal values for the erythrocytes, haemoglobin and haematocrit in horses consuming pellets with spores were recorded, and lower than normal in the other groups. Sensitization of horses to the fungal species was disproven. It is concluded that feeding horses with pellets industrially manufactured with fungal spores represents a very useful tool to implement an integrated control of helminths affecting horses

Direct analysis and identification of pathogenic Lichtheimia species by matrix-assisted laser desorption ionization-time of flight analyzer-mediated mass spectrometry.

PubMed

Schrödl, Wieland; Heydel, Tilo; Schwartze, Volker U; Hoffmann, Kerstin; Grosse-Herrenthey, Anke; Walther, Grit; Alastruey-Izquierdo, Ana; Rodriguez-Tudela, Juan Luis; Olias, Philipp; Jacobsen, Ilse D; de Hoog, G Sybren; Voigt, Kerstin

2012-02-01

Zygomycetes of the order Mucorales can cause life-threatening infections in humans. These mucormycoses are emerging and associated with a rapid tissue destruction and high mortality. The resistance of Mucorales to antimycotic substances varies between and within clinically important genera such as Mucor, Rhizopus, and Lichtheimia. Thus, an accurate diagnosis before onset of antimycotic therapy is recommended. Matrix-assisted laser desorption ionization (MALDI)-time of flight (TOF) mass spectrometry (MS) is a potentially powerful tool to rapidly identify infectious agents on the species level. We investigated the potential of MALDI-TOF MS to differentiate Lichtheimia species, one of the most important agents of mucormycoses. Using the Bruker Daltonics FlexAnalysis (version 3.0) software package, a spectral database library with m/z ratios of 2,000 to 20,000 Da was created for 19 type and reference strains of clinically relevant Zygomycetes of the order Mucorales (12 species in 7 genera). The database was tested for accuracy by use of 34 clinical and environmental isolates of Lichtheimia comprising a total of five species. Our data demonstrate that MALDI-TOF MS can be used to clearly discriminate Lichtheimia species from other pathogenic species of the Mucorales. Furthermore, the method is suitable to discriminate species within the genus. The reliability and robustness of the MALDI-TOF-based identification are evidenced by high score values (above 2.3) for the designation to a certain species and by moderate score values (below 2.0) for the discrimination between clinically relevant (Lichtheimia corymbifera, L. ramosa, and L. ornata) and irrelevant (L. hyalospora and L. sphaerocystis) species. In total, all 34 strains were unequivocally identified by MALDI-TOF MS with score values of >1.8 down to the generic level, 32 out of 34 of the Lichtheimia isolates (except CNM-CM 5399 and FSU 10566) were identified accurately with score values of >2 (probable species

Biomolecular characterization, identification, enzyme activities of molds and physiological changes in sweet potatoes (Ipomea batatas) stored under controlled atmospheric conditions

PubMed Central

Oladoye, C. O.; Connerton, I. F.; Kayode, R. M. O.; Omojasola, P. F.; Kayode, I. B.

2016-01-01

Microbial attacks during storage are one of the primary causes of product deterioration, and can limit the process of prolonging the shelf-life of harvested food. In this study, sweet potatoes were stored at temperatures of 13, 21, and 29 °C for 4 weeks. Samples were collected during storage and plated on potato dextrose agar, from which axenic mold cultures were obtained and identified using 26S rRNA gene sequences. Physiological changes of potato tubers were assessed with respect to pathogenicity, enzyme activity, and atmospheric storage conditions. Six fungal species were identified, namely Penicillium chrysogenum (P. rubens), P. brevicompactum, Mucor circinelloides, Cladosporium cladosporiodes, P. expansum, and P. crustosum. The following fungal isolates, namely P. expansum, P. brevicompactum, and Rhizopus oryzae, were recovered from the re-infected samples and selected according to their levels of enzyme activity. This study revealed high levels of activity for cellulase and pectinase, which were most notable during the initial three days of testing, and were followed by a steady decrease (P<0.05). Polygalacturonase activity was prominent with values ranging from 12.64 to 56.79 U/mg (P. expansum) and 18.36 to 79.01 U/mg (P. brevicompactum). Spoilage was obvious in the control group, which had a 100% decay at the end of the experimental period compared with samples treated with iprodione and sodium hypochlorite, in which the decay rates were 5% and 55%, respectively. The data for the iprodione- and sodium hypochlorite-treated samples at the end of the 3-month storage period showed that they were significantly different (P=0.041), with the sodium hypochlorite-treated samples producing twice the rate of infection compared to the iprodione-treated samples. The comparative rate of the progression of decay in the treated samples can be expressed as iprodione

Direct Analysis and Identification of Pathogenic Lichtheimia Species by Matrix-Assisted Laser Desorption Ionization–Time of Flight Analyzer-Mediated Mass Spectrometry

PubMed Central

Schrödl, Wieland; Heydel, Tilo; Schwartze, Volker U.; Hoffmann, Kerstin; Große-Herrenthey, Anke; Walther, Grit; Alastruey-Izquierdo, Ana; Rodriguez-Tudela, Juan Luis; Olias, Philipp; Jacobsen, Ilse D.; de Hoog, G. Sybren

2012-01-01

Zygomycetes of the order Mucorales can cause life-threatening infections in humans. These mucormycoses are emerging and associated with a rapid tissue destruction and high mortality. The resistance of Mucorales to antimycotic substances varies between and within clinically important genera such as Mucor, Rhizopus, and Lichtheimia. Thus, an accurate diagnosis before onset of antimycotic therapy is recommended. Matrix-assisted laser desorption ionization (MALDI)–time of flight (TOF) mass spectrometry (MS) is a potentially powerful tool to rapidly identify infectious agents on the species level. We investigated the potential of MALDI-TOF MS to differentiate Lichtheimia species, one of the most important agents of mucormycoses. Using the Bruker Daltonics FlexAnalysis (version 3.0) software package, a spectral database library with m/z ratios of 2,000 to 20,000 Da was created for 19 type and reference strains of clinically relevant Zygomycetes of the order Mucorales (12 species in 7 genera). The database was tested for accuracy by use of 34 clinical and environmental isolates of Lichtheimia comprising a total of five species. Our data demonstrate that MALDI-TOF MS can be used to clearly discriminate Lichtheimia species from other pathogenic species of the Mucorales. Furthermore, the method is suitable to discriminate species within the genus. The reliability and robustness of the MALDI-TOF-based identification are evidenced by high score values (above 2.3) for the designation to a certain species and by moderate score values (below 2.0) for the discrimination between clinically relevant (Lichtheimia corymbifera, L. ramosa, and L. ornata) and irrelevant (L. hyalospora and L. sphaerocystis) species. In total, all 34 strains were unequivocally identified by MALDI-TOF MS with score values of >1.8 down to the generic level, 32 out of 34 of the Lichtheimia isolates (except CNM-CM 5399 and FSU 10566) were identified accurately with score values of >2 (probable species

Effects of Soil Pre-Treatment with Basamid® Granules, Brassica juncea, Raphanus sativus, and Tagetes patula on Bacterial and Fungal Communities at Two Apple Replant Disease Sites

PubMed Central

Yim, Bunlong; Nitt, Heike; Wrede, Andreas; Jacquiod, Samuel; Sørensen, Søren J.; Winkelmann, Traud; Smalla, Kornelia

2017-01-01

Nurseries producing apple and rose rootstock plants, apple orchards as well as rose production often experience replanting problems after several cultivations at the same site when a chemical soil disinfectant is not applied. The etiology of apple and rose replanting problems is most likely caused by soil-borne pathogen complex, defined as “replant disease (RD)”. Symptoms typical of RD are reduced shoot and root growth, a smaller leaf area, a significant decrease in plant biomass, yield and fruit quality and a shorter life span. In our previous study, we showed that RD symptoms were reduced when apple rootstock M106 were grown in RD soils treated either with the soil fumigant Basamid or after biofumigation by incorporating Brassica juncea or Raphanus sativus or by growing Tagetes under field conditions compared to untreated control soil. The present study aimed at identifying potential bacterial and fungal taxa that were affected by different soil treatments and linking bacterial and fungal responders to plant performance. Miseq® Illumina® sequencing of 16S rRNA gene fragments (bacteria) and ITS regions (fungi) amplified from total community DNA extracted from soil samples taken 4 weeks after treatments were performed. Soil properties and culture history of the two RD sites greatly influenced soil microbiomes. Several bacterial genera were identified that significantly increased in treated soils such as Arthrobacter (R. sativus, both sites), Curtobacterium (Basamid, both sites), Terrimonas (Basamid and R. sativus, site A) and Ferruginibacter (B. juncea, site K and R. sativus, site A) that were also significantly and positively correlated with growth of apple M106 plants. Only few fungal genera, such as Podospora, Monographella and Mucor, were significantly promoted in soils treated with B. juncea and R. sativus (both sites). The least pronounced changes were recorded for bacterial as well as fungal communities in the RD soils planted with Tagetes. The

Microbiological contamination with moulds in work environment in libraries and archive storage facilities.

PubMed

Zielinska-Jankiewicz, Katarzyna; Kozajda, Anna; Piotrowska, Malgorzata; Szadkowska-Stanczyk, Irena

2008-01-01

Microbiological contamination with fungi, including moulds, can pose a significant health hazard to those working in archives or museums. The species involved include Aspergillus, Penicillium, Geotrichum, Alternaria, Cladosporium, Mucor, Rhizopus, Trichoderma, Fusarium which are associated mostly with allergic response of different types. The aim of the study was to analyse, both in quantitative and qualitative terms, workplace air samples collected in a library and archive storage facilities. Occupational exposure and the related health hazard from microbiological contamination with moulds were assessed in three archive storage buildings and one library. Air samples (total 60) were collected via impact method before work and at hourly intervals during work performance. Surface samples from the artifacts were collected by pressing a counting (RODAC) plate filled with malt extract agar against the surface of the artifacts. The air sample and surface sample analyses yielded 36 different mould species, classified into 19 genera, of which Cladosporium and Penicillium were the most prevalent. Twelve species were regarded as potentially pathogenic for humans: 8 had allergic and 11 toxic properties, the latter including Aspergillus fumigatus. Quantitative analysis revealed air microbiological contamination with moulds at the level ranging from 1.8 x 10(2)-2.3 x 10(3) cfu/m(3). In surface samples from library and archive artifacts, 11 fungal species were distinguished; the number of species per artifact varying from 1-6 and colony count ranging from 4 x 10(1) to 8-10(1) cfu/100 cm(2). Higher contamination levels were found only for Cladosporium cladosporioides (1.48 x 10(3) cfu/100 cm(2)) and Paecillomyces varioti (1.2 x 10(2) cfu/100 cm(2)). At the workposts examined, although no clearly visible signs of mould contamination could be found, the study revealed abundant micromycetes, with the predominant species of Cladosporium and Penicillium. The detected species included

[Survey on fungi contamination and natural occurrence of mycotoxins in 94 corn feed ingredients collected from China].

PubMed

Han, X M; Zhang, H Y; Zhang, J; Xu, W J; Liu, D; Jiang, T; Xu, J; Li, F Q

2016-10-06

Objective: To investigate fungi contamination and the natural occurrence of mycotoxins in corn feed ingredients collected from China. Methods: A total of 94 corn feed ingredient samples were collected from 8 Chinese provinces(i.e., Anhui, Hebei, Heilongjiang, Jilin, Jiangsu, Liaoning, Inner Mongolia, and Shandong)in February 2014. A tandem ultra-performance liquid chromatographymass spectrometry method was used for simultaneous detection of twelve kinds of mycotoxins, including aflatoxin(AF), type A and type B tricothecenes, and zearalenone(ZEN). Contaminated fungi were also identified and counted. Results: AF was detected in 36.2%(34/94)of samples; the concentration of AFB 1 was the highest in the four AFs with the range: 0.3~181.3 μg/kg; and then followed by AFB 2 (range: 1.0-74.3 μg/kg). There were 7 samples(7.5%)with AFB 1 concentrations higher than the tolerance limit of 50 μg/kg. The concentration of type A tricothecenes in all samples was lower(0.1-10.5 μg/kg). DON had the most serious contamination than other kind of type B tricothecenes(range: 0.7-606.6 μg/kg; median: 66.3 μg/kg). The DON concentration in all samples was below the tolerance limit of 1 000 μg/kg. ZEN was detected in 76.6%(72/ 94)of samples(median: 36.9 μg/kg), with 3 samples having ZEN concentrations higher than the tolerance limit of 500 μg/kg. The survey on fungi contamination showed that all samples were contaminated by fungi(range: 5.0-1.4×10 5 CFU/g). There were 18 and 3 samples with quantities of fungi higher than the tolerance and forbidden limits, respectively. The Aspergillus , Penicillium , Fusarium , Trichoderma and Mucor genuses were the predominant fungi in corn feed ingredients, with detection rates of 71.3%(67), 60.6%(57), 71.3%(67), 27.7%(26), and 24.5%(23), respectively. The detection rate of Fusarium moniliforme , 73.4%(69/94)was higher than that of Aspergillus flavus , 41.5%(39/94). Conclusion: In this survey, the corn feed ingredients were not seriously

Comparative "in vitro" evaluation of the antiresorptive activity residing in four Ayurvedic medicinal plants. Hemidesmus indicus emerges for its potential in the treatment of bone loss diseases.

PubMed

Di Pompo, Gemma; Poli, Ferruccio; Mandrone, Manuela; Lorenzi, Beatrice; Roncuzzi, Laura; Baldini, Nicola; Granchi, Donatella

2014-06-11

Four Indian plants, traditionally used in Ayurvedic medicine: Asparagus racemosus Willd., Emblica officinalis Gaertn., Hemidesmus indicus R. Br., and Rubia cordifolia L. were selected on the basis of their ethnobotanical use and of scientific evidence that suggests a potential efficacy in the treatment of bone-loss diseases. The antiresorptive properties of the four plants have been investigated. The aim was to provide adequate evidence for the exploitation of natural compounds as alternative therapeutics for the treatment of diseases caused by increased osteoclast activity. Decoctions were prepared from dried plant material according to the traditional procedure and standardization by HPLC was performed using marker compounds for each species. Total polyphenols, flavonoids and radical scavenging activity of the decoctions were also determined. The bioactivity of the plant decoctions was evaluated in subsequent phases. (1) A cytotoxicity screening was performed on the mouse monocytic RAW 264.7 cell line to define the concentrations that could be utilized in the following step. (2) The antiresorptive properties of plant decoctions were compared with that of a "gold standard" drug (alendronate) by measuring osteoclastogenesis inhibition and osteoclast apoptosis. (3) The toxic effect on bone forming cells was excluded by evaluating the impact on the proliferation of osteogenic precursors (mesenchymal stem cells, MSC). All the decoctions inhibited osteoclastogenesis similarly to alendronate at the highest doses, but Hemidesmus indicus and Rubia cordifolia were also effective at lower concentrations. Apoptosis increased significantly when cells were exposed to the highest concentration of Emblica officinalis, Hemidesmus indicus, and Rubia cordifolia. All concentrations of Emblica officinalis tested inhibited the proliferation of osteogenic precursors, while only the highest doses of Asparagus racemosus and Rubia cordifolia were toxic. On the contrary, Hemidesmus indicus

Medico-botanical study of Yercaud hills in the eastern Ghats of Tamil Nadu, India.

PubMed

Parthipan, M; Aravindhan, V; Rajendran, A

2011-04-01

The study reports medicinal plant survey was conceded in Yercaud hills ranges of Eastern Ghats, Tamil Nadu, India. The study primarily based on field surveys conducted throughout the hills, where dwellers provided information on plant species used as medicine, plant parts used to prepare the remedies and ailments to which the remedies were prescribed. The study resulted about 48- plant species belonging to 45- genera and 29- families of medicinal plants related to folk medicine used by the local people. Among them the most common plants viz., Asparagus racemosus Willd., Cissus quadrangularis L., Gymnema sylvestre R. Br., Hemidesmus indicus (L.) R. Br., Justisia adhatoda L., Ocimum sanctum L., Phyllanthes amarus Schum. & Thonn., Piper nigrum L., Solanum nigrum L., Tinospora cordifolia (Thunb.) Miers, Tridax procumbens L. and Zingiber officinale Roscoe which are used in their daily life to cure various ailments.

Conservation of Mannan Synthesis in Fungi of the Zygomycota and Ascomycota Reveals a Broad Diagnostic Target.

PubMed

Burnham-Marusich, Amanda R; Hubbard, Breeana; Kvam, Alexander J; Gates-Hollingsworth, Marcellene; Green, Heather R; Soukup, Eric; Limper, Andrew H; Kozel, Thomas R

2018-01-01

Ascomycetes and zygomycetes account for the majority of (i) fungi responsible for cutaneous, subcutaneous, and invasive human fungal infections, (ii) plant fungal pathogens, (iii) fungi that threaten global biodiversity, (iv) fungal agents of agricultural spoilage, and (v) fungi in water-damaged buildings. Rapid recognition of fungal infection (or contamination) enables early treatment (or remediation). A bioinformatics search found homologues of Saccharomyces cerevisiae Mnn9p present in members of the Zygomycota and Ascomycota phyla and absent in members of the Chytridiomycota and Basidiomycota. Mnn9p is a component of the yeast mannan polymerization complex and is necessary for α-1,6 mannan production. A monoclonal antibody (2DA6) was produced that was reactive with purified mannans of Mucor , Rhizopus , Aspergillus , Fusarium , and Candida species. Experimentation using a 2DA6 antigen capture enzyme-linked immunosorbent assay (ELISA) and extracts of fungi from the four phyla found agreement between the presence or absence of Mnn9p homologues and production or lack of production of mannan reactive with 2DA6. Studies of cell extracts from yeast mannan mutants identified α-1,6 mannan as the epitope recognized by 2DA6. To translate this finding into a point-of-use diagnostic, a 2DA6 lateral flow immunoassay was constructed that detected mannan in (i) extracts of dermatophytes and fungi that produce trauma-related infection and (ii) tissue from plants infected with Grosmannia clavigera or Sclerotium cepivorum These studies (i) revealed that the conservation of α-1,6-linked mannan in fungi of the Zygomycota and Ascomycota can be exploited as a broad diagnostic target and (ii) have provided a means to detect that target in an immunoassay platform that is well suited for clinic or field use. IMPORTANCE A key question asked when faced with an infection, an infestation, or environmental damage is whether it is a fungus. Identification of fungi as the cause of the

Conservation of Mannan Synthesis in Fungi of the Zygomycota and Ascomycota Reveals a Broad Diagnostic Target

PubMed Central

Hubbard, Breeana; Kvam, Alexander J.; Gates-Hollingsworth, Marcellene; Green, Heather R.; Soukup, Eric; Limper, Andrew H.; Kozel, Thomas R.

2018-01-01

ABSTRACT Ascomycetes and zygomycetes account for the majority of (i) fungi responsible for cutaneous, subcutaneous, and invasive human fungal infections, (ii) plant fungal pathogens, (iii) fungi that threaten global biodiversity, (iv) fungal agents of agricultural spoilage, and (v) fungi in water-damaged buildings. Rapid recognition of fungal infection (or contamination) enables early treatment (or remediation). A bioinformatics search found homologues of Saccharomyces cerevisiae Mnn9p present in members of the Zygomycota and Ascomycota phyla and absent in members of the Chytridiomycota and Basidiomycota. Mnn9p is a component of the yeast mannan polymerization complex and is necessary for α-1,6 mannan production. A monoclonal antibody (2DA6) was produced that was reactive with purified mannans of Mucor, Rhizopus, Aspergillus, Fusarium, and Candida species. Experimentation using a 2DA6 antigen capture enzyme-linked immunosorbent assay (ELISA) and extracts of fungi from the four phyla found agreement between the presence or absence of Mnn9p homologues and production or lack of production of mannan reactive with 2DA6. Studies of cell extracts from yeast mannan mutants identified α-1,6 mannan as the epitope recognized by 2DA6. To translate this finding into a point-of-use diagnostic, a 2DA6 lateral flow immunoassay was constructed that detected mannan in (i) extracts of dermatophytes and fungi that produce trauma-related infection and (ii) tissue from plants infected with Grosmannia clavigera or Sclerotium cepivorum. These studies (i) revealed that the conservation of α-1,6-linked mannan in fungi of the Zygomycota and Ascomycota can be exploited as a broad diagnostic target and (ii) have provided a means to detect that target in an immunoassay platform that is well suited for clinic or field use. IMPORTANCE A key question asked when faced with an infection, an infestation, or environmental damage is whether it is a fungus. Identification of fungi as the cause of the

In vitro antimicrobial activity of ten medicinal plants against clinical isolates of oral cancer cases

PubMed Central

2011-01-01

Background Suppression of immune system in treated cancer patients may lead to secondary infections that obviate the need of antibiotics. In the present study, an attempt was made to understand the occurrence of secondary infections in immuno-suppressed patients along with herbal control of these infections with the following objectives to: (a) isolate the microbial species from the treated oral cancer patients along with the estimation of absolute neutrophile counts of patients (b) assess the in vitro antimicrobial activity medicinal plants against the above clinical isolates. Methods Blood and oral swab cultures were taken from 40 oral cancer patients undergoing treatment in the radiotherapy unit of Regional Cancer Institute, Pt. B.D.S. Health University, Rohtak, Haryana. Clinical isolates were identified by following general microbiological, staining and biochemical methods. The absolute neutrophile counts were done by following the standard methods. The medicinal plants selected for antimicrobial activity analysis were Asphodelus tenuifolius Cav., Asparagus racemosus Willd., Balanites aegyptiaca L., Cestrum diurnum L., Cordia dichotoma G. Forst, Eclipta alba L., Murraya koenigii (L.) Spreng. , Pedalium murex L., Ricinus communis L. and Trigonella foenum graecum L. The antimicrobial efficacy of medicinal plants was evaluated by modified Kirby-Bauer disc diffusion method. MIC and MFC were investigated by serial two fold microbroth dilution method. Results Prevalent bacterial pathogens isolated were Staphylococcus aureus (23.2%), Escherichia coli (15.62%), Staphylococcus epidermidis (12.5%), Pseudomonas aeruginosa (9.37%), Klebsiella pneumonia (7.81%), Proteus mirabilis (3.6%), Proteus vulgaris (4.2%) and the fungal pathogens were Candida albicans (14.6%), Aspergillus fumigatus (9.37%). Out of 40 cases, 35 (87.5%) were observed as neutropenic. Eight medicinal plants (A. tenuifolius, A. racemosus, B. aegyptiaca, E. alba, M. koenigii, P. murex R. communis and T

In vitro antimicrobial activity of ten medicinal plants against clinical isolates of oral cancer cases.

PubMed

Panghal, Manju; Kaushal, Vivek; Yadav, Jaya P

2011-05-20

Suppression of immune system in treated cancer patients may lead to secondary infections that obviate the need of antibiotics. In the present study, an attempt was made to understand the occurrence of secondary infections in immuno-suppressed patients along with herbal control of these infections with the following objectives to: (a) isolate the microbial species from the treated oral cancer patients along with the estimation of absolute neutrophile counts of patients (b) assess the in vitro antimicrobial activity medicinal plants against the above clinical isolates. Blood and oral swab cultures were taken from 40 oral cancer patients undergoing treatment in the radiotherapy unit of Regional Cancer Institute, Pt. B.D.S. Health University,Rohtak, Haryana. Clinical isolates were identified by following general microbiological, staining and biochemical methods. The absolute neutrophile counts were done by following the standard methods. The medicinal plants selected for antimicrobial activity analysis were Asphodelus tenuifolius Cav., Asparagus racemosus Willd., Balanites aegyptiaca L., Cestrum diurnum L., Cordia dichotoma G. Forst, Eclipta alba L., Murraya koenigii (L.) Spreng. , Pedalium murex L., Ricinus communis L. and Trigonella foenum graecum L. The antimicrobial efficacy of medicinal plants was evaluated by modified Kirby-Bauer disc diffusion method. MIC and MFC were investigated by serial two fold microbroth dilution method. Prevalent bacterial pathogens isolated were Staphylococcus aureus (23.2%), Escherichia coli (15.62%), Staphylococcus epidermidis (12.5%), Pseudomonas aeruginosa (9.37%), Klebsiella pneumonia (7.81%), Proteus mirabilis (3.6%), Proteus vulgaris (4.2%) and the fungal pathogens were Candida albicans (14.6%), Aspergillus fumigatus (9.37%). Out of 40 cases, 35 (87.5%) were observed as neutropenic. Eight medicinal plants (A. tenuifolius, A. racemosus, B. aegyptiaca, E. alba, M. koenigii, P. murex R. communis and T. foenum graecum) showed

Investigation of neuropsychopharmacological effects of a polyherbal formulation on the learning and memory process in rats.

PubMed

Shah, Js; Goyal, Rk

2011-04-01

To investigate the neuropsychopharmacological effect of a polyherbal formulation (PHF) on the learning and memory processes in rats. PHF contains Withania somnifera (Ashwagandha), Nardostachys jatamansi (Jatamansi), Rauwolfia serpentina (Sarpagandha), Evolvulus alsinoides (Shankhpushpi), Asparagus racemosus (Shatavari), Emblica officinalis (Amalki), Mucuna pruriens (Kauch bij extract), Hyoscyamus niger (Khurasani Ajmo), Mineral resin (Shilajit), Pearl (Mukta Shukhti Pishti), and coral calcium (Praval pishti). Its effect (500 mg / kg, p.o.) on the learning and memory processes was tested. The activity of PHF on memory acquisition and retention was studied using passive avoidance learning and elevated plus maze model (EPM) in rats. The animals treated with PHF showed a significant decrease in transfer latency as compared to the control group in EPM. PHF also produced significant improvement in passive avoidance acquisition and memory retrieval, as compared to the controls and reduced the latency to reach the shock free zone (SFZ) after 24 hours. The PHF produces significant improvement in passive avoidance acquisition and memory retrieval in rats, which needs further investigation.

Zygomycetes in Human Disease

PubMed Central

Ribes, Julie A.; Vanover-Sams, Carolyn L.; Baker, Doris J.

2000-01-01

The Zygomycetes represent relatively uncommon isolates in the clinical laboratory, reflecting either environmental contaminants or, less commonly, a clinical disease called zygomycosis. There are two orders of Zygomycetes containing organisms that cause human disease, the Mucorales and the Entomophthorales. The majority of human illness is caused by the Mucorales. While disease is most commonly linked to Rhizopus spp., other organisms are also associated with human infection, including Mucor, Rhizomucor, Absidia, Apophysomyces, Saksenaea, Cunninghamella, Cokeromyces, and Syncephalastrum spp. Although Mortierella spp. do cause disease in animals, there is no longer sufficient evidence to suggest that they are true human pathogens. The spores from these molds are transmitted by inhalation, via a variety of percutaneous routes, or by ingestion of spores. Human zygomycosis caused by the Mucorales generally occurs in immunocompromised hosts as opportunistic infections. Host risk factors include diabetes mellitus, neutropenia, sustained immunosuppressive therapy, chronic prednisone use, iron chelation therapy, broad-spectrum antibiotic use, severe malnutrition, and primary breakdown in the integrity of the cutaneous barrier such as trauma, surgical wounds, needle sticks, or burns. Zygomycosis occurs only rarely in immunocompetent hosts. The disease manifestations reflect the mode of transmission, with rhinocerebral and pulmonary diseases being the most common manifestations. Cutaneous, gastrointestinal, and allergic diseases are also seen. The Mucorales are associated with angioinvasive disease, often leading to thrombosis, infarction of involved tissues, and tissue destruction mediated by a number of fungal proteases, lipases, and mycotoxins. If the diagnosis is not made early, dissemination often occurs. Therapy, if it is to be effective, must be started early and requires combinations of antifungal drugs, surgical intervention, and reversal of the underlying risk

Investigation of Neuropsychopharmacological Effects of a Polyherbal Formulation on the Learning and Memory Process in Rats

PubMed Central

Shah, JS; Goyal, RK

2011-01-01

Objective: To investigate the neuropsychopharmacological effect of a polyherbal formulation (PHF) on the learning and memory processes in rats. Materials and Methods: PHF contains Withania somnifera (Ashwagandha), Nardostachys jatamansi (Jatamansi), Rauwolfia serpentina (Sarpagandha), Evolvulus alsinoides (Shankhpushpi), Asparagus racemosus (Shatavari), Emblica officinalis (Amalki), Mucuna pruriens (Kauch bij extract), Hyoscyamus niger (Khurasani Ajmo), Mineral resin (Shilajit), Pearl (Mukta Shukhti Pishti), and coral calcium (Praval pishti). Its effect (500 mg / kg, p.o.) on the learning and memory processes was tested. The activity of PHF on memory acquisition and retention was studied using passive avoidance learning and elevated plus maze model (EPM) in rats. Results: The animals treated with PHF showed a significant decrease in transfer latency as compared to the control group in EPM. PHF also produced significant improvement in passive avoidance acquisition and memory retrieval, as compared to the controls and reduced the latency to reach the shock free zone (SFZ) after 24 hours. Conclusion: The PHF produces significant improvement in passive avoidance acquisition and memory retrieval in rats, which needs further investigation. PMID:21731356

[Identification of filamentous fungi isolated from clinical samples by two different methods and their susceptibility results].

PubMed

Direkel, Sahin; Otağ, Feza; Aslan, Gönül; Ulger, Mahmut; Emekdaş, Gürol

2012-01-01

Molds are widely distributed in nature. Aspergillus spp. represent the most frequently observed causative agents, however less frequent pathogens Fusarium, Scedosporium and Zygomycetes have also been considered the most important causes of morbidity and mortality in profoundly immunosuppressed hosts. The aims of this study were to identify filamentous fungi isolated from clinical specimens by conventional and molecular methods, and to detect their antifungal susceptibilities. A total of 6742 clinical specimens obtained from hospitalized patients at critical units of Mersin University Medical Faculty Hospital and sent to our laboratory between April 2008-January 2010 were included in the study. The isolates were identified by classical mycological methods and polymerase chain reaction-based DNA sequencing. Susceptibilities to fluconazole and voriconazole were tested by disk diffusion method and to fluconazole, voriconazole, amfoterisin B, caspofungin and posaconazole by E-test. Filamentous fungi were isolated from 71 (1.05%) samples (13 sputum, 4 wound, 4 peritoneal fluid, 3 extrenal ear discharge, 3 abscess and one of each cerebrospinal fluid, blood, tissue biopsy, nasal swab and conjunctival swab) which belonged to 32 patients (13 female, 19 male; age range 7 months-77 years, mean age: 46.6 years). Of the patients 62.3% presented one or more risk factors such as chronic renal failure (n= 8), chronic obstructive lung disease (n= 6), malignancy (n= 6), diabetes mellitus (n= 5) and peripheral vascular disease (n= 5). Of the isolates six were identified as Aspergillus niger, six as Aspergillus flavus, five as Aspergillus fumigatus, four as Aspergillus terreus, five as Fusarium spp., two as Bipolaris spp., and one of each as Acremonium spp., Aurebasidium spp., Mucor spp., and Scedosporium spp. By conventional methods. Three isolates exhibited different identities by DNA sequencing. All Aspergillus isolates were correctly identified at species level by both methods

Neuroinfections caused by fungi.

PubMed

Góralska, Katarzyna; Blaszkowska, Joanna; Dzikowiec, Magdalena

2018-05-21

Fungal infections of the central nervous system (FIs-CNS) have become significantly more common over the past 2 decades. Invasion of the CNS largely depends on the immune status of the host and the virulence of the fungal strain. Infections with fungi cause a significant morbidity in immunocompromised hosts, and the involvement of the CNS may lead to fatal consequences. One hundred and thirty-five articles on fungal neuroinfection in PubMed, Google Scholar, and Cochrane databases were selected for review using the following search words: "fungi and CNS mycoses", CNS fungal infections", "fungal brain infections", " fungal cerebritis", fungal meningitis", "diagnostics of fungal infections", and "treatment of CNS fungal infections". All were published in English with the majority in the period 2000-2018. This review focuses on the current knowledge of the epidemiology, clinical presentations, diagnosis, and treatment of selected FIs-CNS. The FIs-CNS can have various clinical presentations, mainly meningitis, encephalitis, hydrocephalus, cerebral abscesses, and stroke syndromes. The etiologic factors of neuroinfections are yeasts (Cryptococcus neoformans, Candida spp., Trichosporon spp.), moniliaceous moulds (Aspergillus spp., Fusarium spp.), Mucoromycetes (Mucor spp., Rhizopus spp.), dimorphic fungi (Blastomyces dermatitidis, Coccidioides spp., Histoplasma capsulatum), and dematiaceous fungi (Cladophialophora bantiana, Exophiala dermatitidis). Their common route of transmission is inhalation or inoculation from trauma or surgery, with subsequent hematogenous or contiguous spread. As the manifestations of FIs-CNS are often non-specific, their diagnosis is very difficult. A fast identification of the etiological factor of neuroinfection and the application of appropriate therapy are crucial in preventing an often fatal outcome. The choice of effective drug depends on its extent of CNS penetration and spectrum of activity. Pharmaceutical formulations of amphotericin B

Enhanced mucosal immune responses against tetanus toxoid using novel delivery system comprised of chitosan-functionalized gold nanoparticles and botanical adjuvant: characterization, immunogenicity, and stability assessment.

PubMed

Barhate, Ganesh; Gautam, Manish; Gairola, Sunil; Jadhav, Suresh; Pokharkar, Varsha

2014-11-01

Approaches based on combined use of delivery systems and adjuvants are being favored to maximize efficient mucosal delivery of antigens. Here, we describe a novel delivery system comprised of chitosan-functionalized gold nanoparticles (CsAuNPs) and saponin-containing botanical adjuvant; Asparagus racemosus extract (ARE) for oral delivery of tetanus toxoid (TT). A significant increase in TT-specific IgG (34.53-fold) and IgA (43.75-fold) was observed when TT-CsAuNPs were formulated with ARE (TT-ARE-CsAuNPs). The local IgA immune responses for TT also showed a significant increase (106.5-fold in intestine washes and 99.74-fold in feces) with ARE-based formulations as compared with plain TT group. No effect of ARE was observed on size, charge, and loading properties of CsAuNPs. Additionally, no effect of ARE and CsAuNPs was observed on antigenicity and secondary structure of TT as determined by fluorescence, circular dichroism, and Fourier transform infrared spectroscopy. The stability studies demonstrated excellent stability profile of formulation at recommended storage conditions. The study establishes the possible role of immunomodulatory adjuvants in particulate delivery systems for mucosal delivery of vaccines. © 2014 Wiley Periodicals, Inc. and the American Pharmacists Association.

Food and feeding behaviour of Asiatic elephant (Elephas maximus Linn.) in Kuldiha Wild Life Sanctuary, Odisha, India.

PubMed

Mohapatra, Kalpana K; Patra, A K; Paramanik, D S

2013-01-01

The feeding behaviour of Asiatic elephant (Elephas maximus) with food reference was studied in Kuldiha Wildlife Sanctuary in Odisha during 2007 to 2009. Though the study area houses a good number of plant species only 71 species were identified as elephant fodder plants. The food trail of elephant was observed as twig breaking, bark peeling, branch breaking, stem twisting uprooting and flower plucking in different regions of study area during different seasons. Alteration of predominantly browsing strategy with that of grazing around the year was related to seasonal variation of food plants. Consumption of tree species (56%) was highest as compared to shrubs (20%), herbs (14%) and climbers (10%). A high degree of variation in dicot- monocot ratio (61:10)) was marked during identification of elephant fodder plant by direct observation. Microscopic analysis of dung showing a high degree of variation in average dicot- monocot ratio suggested that the food plant selection of elephant was highly opportunistic and seasonal. The elephants extensively fed on the plant species like Careya arborea, Kydia calycina, Helicteres isora, Mallotus philippinensis, Aegle marmelos, Zizyphus mauritiona, Bauhinia racemosa, Bauhinia vahlii, Mimosa pudica, Asparagus racemosus, Smilax zeylanica and Diosporea species. They were fond of Madhuca indica (Mahula) flowers in winter and fruits of Mangifera indica (Mango) in summer. They were never found feeding on Tectona grandis and Eucalyptus maculate inside the study area.

An appraisal of eighteen commonly consumed edible plants as functional food based on their antioxidant and starch hydrolase inhibitory activities.

PubMed

Lee, Yian Hoon; Choo, Candy; Watawana, Mindani I; Jayawardena, Nilakshi; Waisundara, Viduranga Y

2015-11-01

Eighteen edible plants were assessed for their antioxidant potential based on oxygen radical absorbance capacity (ORAC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, total phenolics, vitamin C content and various lipophilic antioxidants. The inhibitory activities of the plant extracts against the enzymatic activities of α-amylase and α-glucosidase were also evaluated. The antioxidant and starch hydrolase activities of the plants varied widely across a single batch of analysis. The ORAC and DPPH radical scavenging EC50 values varied between 298 and 1984 Trolox equivalents g(-1) fresh weight and between 91 and 533 mg kg(-1) fresh weight, respectively. The total phenolics and vitamin C contents varied between 32 and 125 mg gallic acid equivalents g(-1) fresh weight and between 96 and 285 µg g(-1) fresh weight, respectively. All the plants contained neoxanthin, violaxanthin, and α- and β-carotene in varying amounts. Coccinia grandis, Asparagus racemosus, Costus speciosus, Amaranthus viridis and Annona muricata displayed the highest inhibitory activities against starch hydrolases. They were the most efficient against the breakdown of seven starches exposed to the two enzymes as well. Overall, the edible plants were observed to display a high antioxidant potential with starch hydrolase inhibitory properties, which were beneficial in their being recognized as functional food. © 2014 Society of Chemical Industry.

The use of galactogogues in the breastfeeding mother.

PubMed

Forinash, Alicia B; Yancey, Abigail M; Barnes, Kylie N; Myles, Thomas D

2012-10-01

To review data regarding the efficacy of galactogogues available in the US to increase breast milk production in postpartum mothers. Literature was sought using PubMed (1966-June 2012) and EMBASE (1973-June 2012). Search terms included breastfeeding, breast milk, lactation, galactogogue, metoclopramide, oxytocin, fenugreek, milk thistle, silymarin, growth hormone, thyroid releasing hormone, medroxyprogesterone, domperidone, goat's rue, beer, Asparagus racemosus, shatavari, Medicago sativa, alfalfa, Onicus benedictus, blessed thistle, Galega officinalis, brewer's yeast, and herbals. All studies including humans and published in English with data assessing the efficacy of galactogogues for increasing breast milk production were evaluated. Breast milk is considered the optimal food source for newborns through 1 year of age. Many factors influence overall maternal production, including maternal pain, illness, balance of time when returning to work, anxiety, or emotional stress. Although a variety of herbal and pharmaceutical options have anecdotal evidence of their ability to improve breast milk production, peer-reviewed studies proving their efficacy are lacking. Metoclopramide, oxytocin, fenugreek, and milk thistle have shown mixed results in improving milk production; however, the trials were small and had a variety of limitations. Nonpharmacologic recommendations should be exhausted before adding therapy. Although anecdotal evidence encourages the use of metoclopramide, fenugreek, asparagus, and milk thistle for their galactogogue properties, efficacy and safety data in the literature are lacking. Oxytocin and domperidone are potentially available for compounding purposes, but safety data are limited. More studies are needed to evaluate the effects of available galactogogues on breast milk production.

Unified phenology model with Bayesian calibration for several European species in Belgium

NASA Astrophysics Data System (ADS)

Fu, Y. S. H.; Demarée, G.; Hamdi, R.; Deckmyn, A.; Deckmyn, G.; Janssens, I. A.

2009-04-01

Plant phenology is a good bio-indicator for climate change, and this has brought a significant increase of interest. Many kinds of phenology models have been developed to analyze and predict the phenological response to climate change, and those models have been summarized into one kind of unified model, which could be applied to different species and environments. In our study, we selected seven European woody plant species (Betula verrucosa, Quercus robur pedunculata, Fagus sylvatica, Fraxinus excelsior, Symphoricarpus racemosus, Aesculus hippocastanum, Robinia pseudoacacia) occurring in five sites distributed across Belgium. For those sites and tree species, phenological observations such as bud burst were available for the period 1956 - 2002. We also obtained regional downscaled climatic data for each of these sites, and combined both data sets to test the unified model. We used a Bayesian approach to generate distributions of model parameters from the observation data. In this poster presentation, we compare parameter distributions between different species and between different sites for individual species. The results of the unified model show a good agreement with the observations, except for Fagus sylvatica. The failure to reproduce the bud burst data for Fagus sylvatica suggest that the other factors not included in the unified model affect the phenology of this species. The parameter series show differences among species as we expected. However, they also differed strongly for the same species among sites.Further work should elucidate the mechanism that explains why model parameters differ among species and sites.

Ethnobotanical Study of Plants Used in the Management of HIV/AIDS-Related Diseases in Livingstone, Southern Province, Zambia.

PubMed

Chinsembu, Kazhila C

2016-01-01

Faced with critical shortages of staff, long queues, and stigma at public health facilities in Livingstone, Zambia, persons who suffer from HIV/AIDS-related diseases use medicinal plants to manage skin infections, diarrhoea, sexually transmitted infections, tuberculosis, cough, malaria, and oral infections. In all, 94 medicinal plant species were used to manage HIV/AIDS-related diseases. Most remedies are prepared from plants of various families such as Combretaceae, Euphorbiaceae, Fabaceae, and Lamiaceae. More than two-thirds of the plants (mostly leaves and roots) are utilized to treat two or more diseases related to HIV infection. Eighteen plants, namely, Achyranthes aspera L., Lannea discolor (Sond.) Engl., Hyphaene petersiana Klotzsch ex Mart., Asparagus racemosus Willd., Capparis tomentosa Lam., Cleome hirta Oliv., Garcinia livingstonei T. Anderson, Euclea divinorum Hiern, Bridelia cathartica G. Bertol., Acacia nilotica Delile, Piliostigma thonningii (Schumach.) Milne-Redh., Dichrostachys cinerea (L.) Wight and Arn., Abrus precatorius L., Hoslundia opposita Vahl., Clerodendrum capitatum (Willd.) Schumach., Ficus sycomorus L., Ximenia americana L., and Ziziphus mucronata Willd., were used to treat four or more disease conditions. About 31% of the plants in this study were administered as monotherapies. Multiuse medicinal plants may contain broad-spectrum antimicrobial agents. However, since widely used plants easily succumb to the threats of overharvesting, they need special protocols and guidelines for their genetic conservation. There is still need to confirm the antimicrobial efficacies, pharmacological parameters, cytotoxicity, and active chemical ingredients of the discovered plants.

The effect of different land uses on arbuscular mycorrhizal fungi in the northwestern Black Sea Region.

PubMed

Palta, Şahin; Lermi, Ayşe Genç; Beki, Rıdvan

2016-06-01

The object of the present research was to establish correlations between the status of root colonization of arbuscular mycorrhizal fungi (AMF) and different types of land use. In order to achieve this aim, rhizosphere soil samples from grassland crops were taken during June and July of 2013 in order to use for determining several soil characteristics. The 27 different taxa and 60 soil samples were collected from the rhizosphere level in the study areas. The existence of AMF was confirmed in 100 % of these plants with different rations of colonization (approximately 12-89 %). Bromus racemosus L. (pasture) was the most dense taxon with the percentage of AMF colonization of 88.9 %, and Trifolium pratense L. (forest) was the least dense taxon with the percentage of AMF colonization of 12.2 % (average 52.0 %). As a result of the statistical analysis, a positive relationship was found between the botanical composition of legumes and AMF colonization (r = 0.35; p = 0.006). However, a negative relationship was determined between botanical composition of other plant families and AMF colonization (r = -0.39; p = 0.002). In addition, a positive relationship was defined between soil pH (H2O) and the root colonization of AMF (r = 0.35; p = 0.005). The pasture had the highest mean value of AMF root colonization. However, the pasture and gap in the forest were in the same group, according to the results of the S-N-K test.

Antibacterial activity of selected medicinal plants against multiple antibiotic resistant uropathogens: a study from Kolli Hills, Tamil Nadu, India.

PubMed

Narayanan, A S; Raja, S S S; Ponmurugan, K; Kandekar, S C; Natarajaseenivasan, K; Maripandi, A; Mandeel, Q A

2011-09-01

The increasing incidence of antibiotic resistance among bacterial pathogens necessitates medicinal plants as an alternate therapy in restricting the resistant infectious organisms. In this primitive study, the antibiotic resistance of organisms isolated from urinary tract infected patients was evaluated using the National Committee for Clinical Laboratory Standards (NCCLS) method and Multiple Antibiotic Resistance (MAR) index values, and the MAR values was also calculated for plant extracts. The 10 common medicinal plants collected from Kolli hills, Namakkal, south India were extracted using the chloroform, methanol, acetone, ethanol and saponification procedure. The efficacy of the extracts on the uropathogens was tested by agar disc diffusion method in order to analyse the inhibitory activity of plant extract on the organisms. Azadiracta indica A. Juss., Tinospora cordifolia (Wild.) and Euphorbia hirta Linn. exhibited high inhibitory activity against most of the 11 tested organisms followed by Cassia javanica Linn. and Phyllanthus niruri Linn. The maximum zone size of 46.3 mm was exhibited by methanol extract of P. niruri Linn. against Pseudomonas aeruginosa. Asparagus racemosus Willd. and Eupatorium triplinerve Vahl had the least activity against resistant pathogens. Saponified lipids of most of the plants exhibited maximum antibacterial activity. Among the tested organisms, P. aeruginosa and Staphylococcus epidermidis were the most susceptible and Serratia marcescens, Enterobacter cloaceae, Citrobacter koseri, and Citrobacter freundii were the least inhibited by most of the extracts of medicinal plants. It is concluded that revised antibiotic policies and more importantly the development of herbal medicine as an alternative may be incorporated in urological practice.

VOC emission into the atmosphere by trees and leaf litter in Polish forests

NASA Astrophysics Data System (ADS)

Isidorov, V.; Smolewska, M.; Tyszkiewicz, Z.

2009-04-01

of vital activity of fungi which decompose leaf litter. Verification of the hypothesis was carried out in the frames of a long-term experiment on decomposition of Scots pine and common spruce litter. The experiment was conducted in natural conditions by placing litter bags on undisturbed topsoil in a forest area where pine and spruce trees prevailed. Part of the needles placed were taken out of the litter bags on the 30th, 282nd, 490th, 630th and 920th day of the experiment for subsequent isolation of fungi which colonize the litter (overall there were identified 98 species which belong to 48 genera of fungi). The experiment revealed that species composition of the fungi colonizing pine and spruce litter varies considerably. At the early stages of decomposition, dematiceous hypnomycetes, which belongs to Cladosporum, Alternaria genera as well as Hormonem genus, capable of utilizing pectin, were dominant species. At the later stages they were substituted with phycomycetes (Mucor sp.), ascomycetes (Penicillium sp.) and basidiomycetes (Trichderma sp.), the most significant group of "secondary saprophytes" able to carry out biodegradation of polysaccharides and lignin. In volatile emissions of all the 15 species of fungi there were identified 80 VOCs of different classes: terpene hydrocarbons and their oxygenated derivatives, C6-C14 aliphatic and C6-C10 aromatic hydrocarbons, C1-C8 alcohols, C2-C9 carbonyls, esters, furans, and halocarbons. VOC composition was specific for each fungi species and depended on the litter of a particular tree species from which it had been isolated. For instance, the emission rate of terpenes, alcohols and carbonyl compounds by Trichoderma polysporum isolated from pine and spruce litter was 3-5 times different. Differences in composition of VOCs emitted into the gas phase by "primary" and "secondary" fungi species which colonize pine and spruce litter are also discussed. This work was supported by Grant MNiSW N305 067 32/2411.

Green leafy porridges: how good are they in controlling glycaemic response?

PubMed

Anuruddhika Subhashinie Senadheera, Senadheera Pathirannehelage; Ekanayake, Sagarika

2013-03-01

Green leafy porridges made with leaf water extracts, rice and coconut milk are common Sri Lankan dietary remedies for diabetes. Though water and ethanolic extracts of most leaves elicit hypoglycaemic effects, data are not available on the efficacy when leaf extracts are incorporated into porridges. Thus, an effort was made to evaluate the proximate compositions and glycaemic index (GI) of some commonly consumed green leafy porridges. The GI of rice porridge and coconut milk porridge were measured to evaluate the effect of other ingredients other than the leaf extracts. Rice was the main contributor to carbohydrate (56-68% on dry weight) and water was the main component in porridges (89-93%). Fat and total dietary fibre contents ranged between 2.5-27% and 5-10%, respectively. The GI of all porridges was low (GI ≤ 55), except Cassia auriculata which had a high GI of 77 ± 12. The GIs of coconut milk, Aerva lanata, Hemidesmus indicus, Scoparia dulcis, Asparagus racemosus, Cephalandra indica, Cardiospermum halicacabum, Murraya koenigii and Aegle marmelos were 31 ± 5, 32 ± 5, 40 ± 8, 39 ± 8, 37 ± 4, 49 ± 8, 46 ± 8, 44 ± 8 and 50 ± 8, respectively. All porridges had a low or medium glycaemic loads ( ≤ 19). However, peak blood glucose reductions of ≥ 25% were observed in all leafy and coconut milk porridges, except in C. auriculata and Atlantia zeylanica, when compared with the glucose control. Therefore, green leafy porridges, except Cassia, can be recommended as breakfast meals for diabetics due to their low GI, peak blood glucose reduction and presence of other nutrients in green leaves.

Neurocysticercosis.

PubMed

Pittella, J E

1997-01-01

Cysticercosis is an infection caused by Taenia solium larvae (cysticerci). When the cysticercus is lodged in the central nervous system (CNS), the disease is known as neurocysticercosis (NCC). NCC is the most frequent and most widely disseminated human neuroparasitosis. It is endemic in many parts of the world, particularly Latin America, Africa, and Asia, and still relatively frequent in Portugal, Spain and Eastern European countries It is also endemic in developed countries with high rates of immigration from endemic areas. Man may act as an intermediate host after ingestion of mature, viable T. solium eggs via the fecal-oral route. The development of lesions in the brain and leptomeninges, and the consequent of onset of symptoms associated with NCC are mainly due to the host immune-inflammatory response. As long as the cysticercus remains viable, there is relative host immune tolerance. It is only when the parasite dies that massive antigen exposure occurs, with intensification of the immune response/inflammatory reaction and the appearance or worsening of symptoms. NCC can be asymptomatic or cause widely varied clinical manifestations, such as seizures, increased intracranial pressure, ischemic cerebrovascular disease, dementia, and signs of compression of the spinal roots/cord. The combination of two or more symptoms is common. Such clinical polymorphism is determined by 1) the number of lesions (single or multiple cysticerci); 2) the location of CNS lesions (subarachnoid, intracerebral, intraventricular, intramedullary); 3) the type of cysticercus (Cysticercus cellulosae, Cysticercus racemosus); 4) the stage of development and involution of the parasite (vesicular or viable, necrotic, fibrocalcified nodule); and 5) the intensity of the host immune-inflammatory response (no inflammatory reaction, leptomeningitis, encephalitis, granular ependymitis, arteritis).

Preliminary Screening of a Classical Ayurvedic Formulation for Anticonvulsant Activity.

PubMed

Dhar, Arnab; Maurya, Santosh Kumar; Mishra, Ashish; Singh, Gireesh Kumar; Singh, Manoj Kumar; Seth, Ankit

2016-01-01

Epilepsy is a serious and complex central nervous system disorder associated with recurrent episodes of convulsive seizures due to the imbalance between excitatory (glutamatergic) and inhibitory (GABAergic) neurotransmitters level in the brain. The available treatments are neither competent to control the seizures nor prevent progress of disease. Since ages, Herbal medicines have remained important sources of medicines in many parts of world which is evidenced through their uses in traditional systems of medicine i.e. Ayurveda, Siddha, Unani, Homeopathy and Chinese etc. A polyherbal formulation (containing Terminalia chebula Retz., Asparagus racemosus Willd., Embelia ribes Burm. F, Acorus calamus L., Tinospora cordifolia (Willd.) Miers, Convolvulus pluricaulis Choisy, Saussurea lappa C.B.Clarke, Achyranthes aspera L.) is mentioned in Ayurvedic classics Bhaiṣajya Ratnāvali . The aim of the study was to evaluate the anticonvulsant activity of the formulation in Maximum electroshock and Pentylenetetrazole induced convulsions in rats. In the present study, a polyherbal formulation was developed as directed by classical text and evaluated for the anticonvulsant activity using Maximal Electroshock Shock (MES) and Pentylenetetrazole (PTZ) induced convulsions in rats. Statistical comparison was done by one way ANOVA followed by the Tukey's multiple comparison test. The obtained results showed that the PHF had a protective role on epilepsy. Treatment with PHF significantly improves antioxidant enzymes activities of superoxide dismutase (SOD) and glutathione (GSH) levels significantly as compared to controls. PHF also significantly decreased malonaldialdehyde (MDA) levels in the brain. Moreover, it also attenuated the PTZ-induced increase in the activity of GABA-T in the rat brain. These findings suggest that PHF might have possible efficacy in the treatment of epilepsy.

New ethnomedicinal claims from Gujjar and Bakerwals tribes of Rajouri and Poonch districts of Jammu and Kashmir, India.

PubMed

Shah, Anwer; Bharati, Kumar Avinash; Ahmad, Javed; Sharma, M P

2015-05-26

Medicinal plants are frequently employed by Gujjar and Bakerwal tribes in Rajouri and Poonch districts of Jammu and Kashmir, India for treatment of various ailments in humans and livestock. Hence, extensive field work was conducted to document the ethnomedicinal plants used by these tribes. Ninety one key informants were interviewed using both in situ and ex situ type of survey methods. The data were analyzed using user value (UV), informant consensus factor (Fic), fidelity level (FL) and relative frequency of citation (Rfc). A total of 104 species of medicinal plants used in the treatment of 40 different non-communicable ailments with 138 remedies are reported. Cephalanthera longifolia (L.) Fritsch was recorded for the first time with ethnomedicinal uses and the rest of the species were previously reported with different medicinal uses by other tribal people. Out of 138 remedies, 129 were employed for human ailments and the remaining seven were used to treat livestock. Most of the species were harvested for leaves (24 species). Herbs (66 species) were the major life form used for medicinal purpose and the most common method of remedy preparation was decoction/tea (27.8%). The highest use value plant was Verbascum thapsus L. for the treatment of stomachache and snake bite. Plants such as Allium humile Kunth, Angelica glauca Edgew, Arnebia benthamii (Wall. ex G. Don) I.M.Johnst, Asparagus racemosus Willd., Balanophora involucrata Hook. f. & Thomson, C. longifolia (L.) Fritsch, Cuscuta epithymum (L.) L., Geranium wallichianum D. Don ex Sweet, Gloriosa superb L., Habenaria intermedia D. Don, Phyllanthus emblica L., Ramaria Formosa (Pers.) Quel. and V. thapsus L. showing high Rfc and FL values may be studied for associated pharmacological activities. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Physicochemical properties of anti Vibrio harveyi egg yolk antibody (IgY) and its immunological influence in Indian white shrimp Fenneropenaeus indicus.

PubMed

Kumaran, Thankamani; Thirumalaikumar, Eswaramoorthy; Lelin, Chinnadurai; Palanikumar, Pandi; Michaelbabu, Mariavincent; Citarasu, Thavasimuthu

2018-03-01

Edible antibodies specific to host pathogens is an attractive approach to establish protective immunity, especially against gastrointestinal pathogens both in humans and animals. The edible antibody of anti-Vibrio harveyi IgY (anti-V. h IgY) was produced by antigen mixed with immunoadjuvant Asparagus racemosus and Glycine max. Hens were immunized and eggs were collected five weeks after the immunization. Anti-V. harveyi IgY stability in different digestive enzymes such as trypsin and chymotrypsin were evaluated to determine its ability to withstand in the gastrointestinal tract of F. indicus. Specific binding activity and concentration (average 9.5% of total IgY content) of the anti-V. h IgY were determined by the ELISA using V. harveyi antigen. Further the anti-V. h IgY diets including V.h wo, V.h A, V.h G and control diets were fed to F. indicus for 60 days. After 30 and 60 of feeding, group of shrimps were challenged with virulent V. harveyi. After the respective days of feeding, haematological and immunological changes were studied. The parameters including total haemocyte count (THC), coagulase activity, oxyhaemocyanin level, prophenoloxidase, intracellular superoxide anion production, lysozyme, phagocytosis and bacterial agglutinin had significantly (P ≤ .001) increased in the experimental groups in comparission with the control diet fed shrimps. The anti-V. h IgY coated diets helped to reduce the Vibrio load and boosted the immune system in F. indicus's against V. harveyi challenge. The research work shows the potential applications of egg yolk antibodies as anti-bacterial prophylactic uses for infectious diseases and suggests an edible antibody concept as an alternative to conventional antibiotics. Copyright © 2018 Elsevier Ltd. All rights reserved.

Anti-diabetic properties of rice-based herbal porridges in diabetic Wistar rats.

PubMed

Senadheera, Senadheera Pathirannehelage Anuruddhika Subhashinie; Ekanayake, Sagarika; Wanigatunge, Chandanie

2014-10-01

The present study aims to investigate anti-hyperglycaemic, anti-hyperlipidaemic and toxic effects of long-term consumption of selected green leafy porridges in a streptozotocin-induced diabetic Wistar rat model. Porridges made with Asparagus racemosus Willd. (AR), Hemidesmus indicus (L) R. Br. W. T. Aiton (HI), Scoparia dulcis L. (SD) and coconut milk porridge (CM) were incorporated into diets of diabetic Wistar rats. Diabetic control (DM) and normal control groups (NC) were provided with standard rat diet. Fasting blood glucose (FBG), HbA1c , C reactive protein (CRP), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), liver enzymes and creatinine were measured. Feed and water intake among diabetic groups were significantly high when compared with those of NC (p < 0.05). All rats in SD (mean = 39 ± 19 g) and NC (mean = 114 ± 7 g) groups gained weight, whereas most rats in other diabetic groups lost weight. Among the diabetic groups, SD group had the lowest mean FBG, FBG increment percentage (45%) and HbA1c (5.8 ± 2.1). FBG increment percentage and HbA1c of SD group were not significantly different to those of NC (38%; 4.7 ± 0.7) (p > 0.05). Among the diabetic groups, lowest TC (119 ± 20.6 mg/dL) and highest HDL-C (33 ± 6.3 mg/dL) were also detected in SD group. Alanine transaminase and creatinine were not significantly different (p > 0.05) among diabetic groups but significant when compared with those of NC. When compared with those of NC, aspartate transaminase levels were significantly (p < 0.05) high in SD, CM and DM groups. Body weight : liver weight and body weight : pancreas weight ratios and CRP were not significantly different among all groups. The study proved that SD porridge reduced weight loss, elicited hypoglycaemic and hypolipidaemic properties, and caused no toxicity in diabetes-induced Wistar rats. Copyright © 2014 John Wiley & Sons, Ltd.

Ethnomedicinal plants used for treating epilepsy by indigenous communities of sub-Himalayan region of Uttarakhand, India.

PubMed

Sharma, Jyotsana; Gairola, Sumeet; Gaur, R D; Painuli, R M; Siddiqi, T O

2013-10-28

Although many plants are claimed to possess anticonvulsant/antiepileptic (AC/AE) properties, but there is very little information available about plants used by various ethnic communities in different parts of India to treat epilepsy, one of the most common disorders of central nervous system (CNS); this communication provides significant ethnomedicinal information on the plants used by indigenous communities: Bhoxa, Tharu and nomadic Gujjars of sub-Himalayan region, Uttarakhand, India to treat epilepsy, so that it could be used as a baseline data for studying chemical constituents and biological activities of these promising plants. To record herbal preparations used by the presently studied communities for treating epilepsy and discuss AC/AE properties of the recorded plants. Ninety one traditional healers (29 Bhoxa, 35 Tharu and 27 nomadic Gujjars) in sub-Himalayan region of Uttarakhand, India were interviewed to collect information on herbal preparations used by them for treating epilepsy. For each recorded species the use value (UV) and fidelity level (FL) was calculated. A total of 24 plants belonging to 24 genera and 22 families were used by the presently studied communities in 26 formulations to treat epilepsy. According to FL and UV values, most preferred species for the treatment of epilepsy by Bhoxa community are Ricinus communis L. and Datura stramonium L.; by nomadic Gujjar community are Martynia annua L., Bacopa monnieri (L.) Wettst. and Ricinus communis L.; and by Tharu community are Allium sativum L., Asparagus racemosus Willd. and Achyranthes aspera L. Eight plants viz., Allium sativum L., Boerhavia diffusa L., Cassia fistula L., Clerodendrum viscosum Vent., Datura stramonium L., Inula cappa DC., Oroxylum indicum (L.) Kurz and Pavetta indica L. recorded in the present survey have been reported for the first time in treatment of epilepsy by these indigenous communities in India. Five out of these eight newly reported plants viz., Cassia fistula L

Adaptogenic activity of Siotone, a polyherbal formulation of Ayurvedic rasayanas.

PubMed

Bhattacharya, S K; Bhattacharya, A; Chakrabarti, A

2000-02-01

Siotone (ST) is a herbal formulation comprising of Withania somnifera, Ocimum sanctum, Asparagus racemosus, Tribulus terristris and shilajit, all of which are classified in Ayurveda as rasayanas which are reputed to promote physical and mental health, improve defence mechanisms of the body and enhance longevity. These attributes are similar to the modern concept of adaptogenic agents, which are, known to afford protection of the human physiological system against diverse stressors. The present study was undertaken to investigate the adaptogenic activity of ST against chronic unpredictable, but mild, footshock stress induced perturbations in behaviour (depression), glucose metabolism, suppressed male sexual behaviour, immunosuppression and cognitive dysfunction in CF strain albino rats. Gastric ulceration, adrenal gland and spleen weights, ascorbic acid and corticosterone concentrations of adrenal cortex, and plasma corticosterone levels, were used as the stress indices. Panax ginseng (PG) was used as the standard adaptogenic agent for comparison. Additionally, rat brain levels of tribulin, an endogenous endocoid postulated to be involved in stress, were also assessed in terms of endogenous monoamine oxidase (MAO) A and MAOB inhibitory activity. Chronic unpredictable footshock induced marked gastric ulceration, significant increase in adrenal gland weight and plasma corticosterone levels, with concomitant decreases in spleen weight, and concentrations of adrenal gland ascorbic acid and corticosterone. These effects were attenuated by ST (50 and 100 mg/kg, p.o.) and PG (100 mg/kg, p.o.), administered once daily over a period of 14 days, the period of stress induction. Chronic stress also induced glucose intolerance, suppressed male sexual behaviour, induced behavioural depression (Porsolt's swim despair test and learned helplessness test) and cognitive dysfunction (attenuated retention of learning in active and passive avoidance tests), and immunosuppression

Ethnopharmacological survey of medicinal plants used by indigenous and tribal people in Rangamati, Bangladesh.

PubMed

Kadir, Mohammad Fahim; Bin Sayeed, Muhammad Shahdaat; Mia, M M K

2012-12-18

There is very limited information regarding plants used by traditional healers in Rangamati, Bangladesh, for treating general ailments. Current study provides significant ethnopharmacological information, both qualitative and quantitative on medical plants in Rangamati. This study aimed to collect, analyze and evaluate the rich ethnopharmacologic knowledge on medicinal plants in Rangamati and attempted to identify the important species used in traditional medicine. Further analysis was done by comparison of the traditional medicinal use with the available scientific literature data. The field survey was carried out in a period of about one year in Rangamati, Bangladesh. A total of 152 people were interviewed, including Traditional Health Practitioners (THPs) and indigenous people through open-ended and semistructured questionnaire. The collected data were analyzed qualitatively and quantitatively. This ethnomedicinal knowledge was compared against the literature for reports of related uses and studies of phytochemical compounds responsible for respective ailments. A total of 144 species of plants, mostly trees, belonging to 52 families were identified for the treatment of more than 90 types of ailments. These ailments were categorized into 25 categories. Leaves were the most frequently used plant parts and decoction is the mode of preparation of major portions of the plant species. The most common mode of administration was oral ingestion and topical application. Informant consensus factor (Fic) values of the present study reflected the high agreement in the use of plants in the treatment of gastro-intestinal complaints and respiratory problems among the informants. Gastro-intestinal complaint had highest use-reports and 3 species of plants, namely Aegle marmelos (L.) Corr., Ananas comosus (L.) Merr., and Terminalia chebula (Gaertn.) Retz., had the highest fidelity level (FL) of 100%. Asparagus racemosus Willd. and Azadirachta indica A. Juss. showed the highest

Inventorization of some ayurvedic plants and their ethnomedicinal use in Kakrajhore forest area of West Bengal.

PubMed

Biswas, Soumyajit; Shaw, Rupa; Bala, Sanjay; Mazumdar, Asis

2017-02-02

& Young shoots (24%), Leaves (18%), Fruits (13%), and Seeds (10%). The most frequent happening ailments are asthma and bronchitis, constipation and dyspepsia, diarrhea, dysentery, ulcer, rheumatism, arthritis, fistula and piles troubles. Most of the Plants (88%) had more than a single therapeutic use. The most important plant species on the basis of use value were Smilex ovalifolia, Emblica officinalis, Curculigo orchioides, Croton roxburghii, Asparagus racemosus, Ziziphus nummularia, Ichnocarpus frutescens, Cisssus adnata, Buettneria herbacea, Litsea glutinosa, Vernonia anthelmintica and Chlorophytum borivilianum. There was a strong agreement among the informants related to the usage of the plants (ICF 0.65-0.93). As per the standard literature in Ayurveda, maximum numbers of documented species (33) are found to be used for 'digestive system disorder' which is analogous to ethnomedicinal use (30), followed by 'skin related problem' i.e. 13 and 20 respectively. Results show that people living around Kakrajhore forest area hold valuable knowledge of the uses of plant resources and some of the ayurvedic plants represent an important component in local livelihood. Though the Kakrajhore Forest area is rich in medicinal plant resources but the species are susceptible to be endangered by intense exploitation by human communities. More in-depth investigations are required for plants with high UV values on their possible phytochemical and pharmacological activity. Apart from this, sustainable management approach and proper conservation strategy for the area is recommended by involvement of local communities and forest department. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.