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Liquid Cultures (liquid + culture)
Terms modified by Liquid Cultures Selected AbstractsChemInform Abstract: Cadinane-Type Sesquiterpenoids, Strobilols E,K, from the Liquid Culture of Strobilurus ohshimae.CHEMINFORM, Issue 51 2008Fuminori Hiramatsu Abstract ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 200 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a "Full Text" option. The original article is trackable via the "References" option. [source] Accumulation of deoxynivalenol and its 15-acetylated form is significantly modulated by oxidative stress in liquid cultures of Fusarium graminearumFEMS MICROBIOLOGY LETTERS, Issue 1 2006Nadia Ponts Abstract Liquid cultures of Fusarium graminearum were supplemented with H2O2 or other oxidative compounds. The accumulation kinetics of the resulting trichothecenes were monitored. At non-lethal concentrations, the H2O2 treatments modulated toxin accumulation, dependent on the method of supplementation. When H2O2 was added at the same time as the inoculation, higher levels of toxins accumulated 30 days later. Conversely, adding H2O2 2 or 7 days after inoculation had little effect. When H2O2 was added daily over the course of the culture, the accumulation of trichothecenes was rapidly and strongly enhanced. The fungus may adapt to oxidative stress when the first exposure to H2O2 occurs at the beginning of the culture course. The highest toxin levels were measured when the H2O2 was added daily. The importance of the first hours of culture was confirmed: pre-treating conidia with H2O2 does not affect their germination kinetics but leads to a reduction in the yield of trichothecenes 40 days later. The H2O2 regulation of this trichothecene accumulation may be specific, as paraquat, another pro-oxidant compound, inhibits their production. Since H2O2 is a major component of the oxidative burst occurring in pathogen/host interactions, these data support the theory that trichothecenes may act as virulence factors. [source] Unravelling the microbial role in ooid formation , results of an in situ experiment in modern freshwater Lake Geneva in SwitzerlandGEOBIOLOGY, Issue 4 2008K. PLEE ABSTRACT The microbial role in the formation of the cortex of low-Mg calcite freshwater ooids in western part of Lake Geneva in Switzerland has been suggested previously, but not demonstrated conclusively. Early work mostly concentrated in hypersaline milieus, and hence little is known about their genesis in freshwater environments. We designed an in situ experiment to mimic the natural process of low-Mg calcite precipitation. A special device was placed in the ooid-rich bank of the lake. It contained frosted glass (SiO2) slides, while quartz (SiO2) is the most abundant mineral composition of ooid nuclei that acted as artificial substrates to favour microbial colonization. Microscopic inspection of the slides revealed a clear seasonal pattern of carbonate precipitates, which were always closely associated with biofilms that developed on the surface of the frosted slides containing extracellular polymeric substance, coccoid and filamentous cyanobacteria, diatoms and heterotrophic bacteria. Carbonate precipitation peaks during early spring and late summer, and low-Mg calcite crystals mostly occur in close association with filamentous and coccoid cyanobacteria (e.g. Tolypothrix, Oscillatoria and Synechococcus, Anacystis, respectively). Further scanning electron microscope inspection of the samples revealed low-Mg calcite with crystal forms varying from anhedral to euhedral rhombohedra, depending on the seasons. Liquid cultures corroborate the in situ observations and demonstrate that under the same physicochemical conditions the absence of biofilms prevents the precipitation of low-Mg calcite crystals. These results illustrate that biofilms play a substantial role in low-Mg calcite ooid cortex formation. It further demonstrates the involvement of microbes in the early stages of ooid development. Combined with ongoing microbial cultures under laboratory-controlled conditions, the outcome of our investigation favoured the hypothesis of external microbial precipitation of low-Mg calcite as the main mechanism involved in the early stage of ooid formation in freshwater Lake Geneva. [source] Antibiotic-Loaded PLGA Nanofibers for Wound Healing Applications,ADVANCED ENGINEERING MATERIALS, Issue 4 2010David A. Soscia Incorporating antibiotics into biocompatible nanoscale non-woven fibrous mats could provide utility for wound healing applications and for incorporation into wound dressing materials. In this study, the antibiotic chloramphenicol (Cm) was incorporated into electrospun poly(lactic-co-glycolic acid) (PLGA) nanofibers, which were then tested for inhibition of bacterial growth for multiple bacterial species (Escherichia coli, Staphylococcus aureus, Bacillus cereus, Salmonella typhimurium, and Pseudomonas aeruginosa). In addition, the cytotoxicity of Cm-PLGA nanofibers was examined for two types of mammalian cells including mouse embryonic stem cells and fibroblasts. Electrospun PLGA nanofibers containing Cm were able to reduce bacterial growth on solid agar plates for all species except for P. aeruginosa. In liquid culture, Cm-loaded nanofibers inhibited growth for E. coli, B. cereus and S. typhimurium by 93% or greater, while P. aeruginosa and S. aureus growth was inhibited by 42% and 56%, respectively. Cm-loaded nanofibers showed limited cytoxicity on fibroblasts and embryonic stem cells, with viability greater than 96% for all conditions tested. These results suggest that Cm can be successfully incorporated into electrospun nanofibers and that these fibers could be used for wound healing applications with minimal cytotoxicity to the surrounding tissue. [source] Mycelium cultivation, chemical composition and antitumour activity of a Tolypocladium sp. fungus isolated from wild Cordyceps sinensisJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2006P.H. Leung Abstract Aims:, To examine and illustrate the morphological characteristics and growth kinetics of Cs-HK1, a Tolypocladium fungus, isolated from wild Cordyceps sinensis in solid and liquid cultures, and the major chemical constituents and antitumour effects of Cs-HK1 mycelium. Methods and Results:, The Cs-HK1 fungus was isolated from the fruiting body of a wild C. sinensis and identified as a Tolypocladium sp. fungus. It grew rapidly at 22,25°C on a liquid medium containing glucose, yeast extract, peptone and major inorganic salts, with a specific growth rate of 1·1 day,1, reaching a cell density of 23·0 g dw l,1 in 7,9 days. Exopolysaccharides accumulated in the liquid culture to about 0·3 g l,1 glucose equivalent. In comparison with natural C. sinensis, the fungal mycelium had similar contents of protein (11·7,,g) and carbohydrate (654·6,,g) but much higher contents of polysaccharide (244·2 mg vs 129·5 mg), adenosine (1116·8,,g vs 264·6 ,g) and cordycepin (65·7 ,g vs 20·8 ,g) (per gram dry weight). Cyclosporin A, an antibiotic commonly produced by Tolypocladium sp., was also detected from the mycelium extract. The hot water extract of mycelium showed low cytotoxic effect on B16 melanoma cells in culture (about 25% inhibition) but significant antitumour effect in animal tests, causing 50% inhibition of B16 cell-induced tumour growth in mice. Conclusions:, The Tolypocladium sp. fungus, Cs-HK1, can be easily cultivated by liquid fermentation. The mycelium biomass contained the major bioactive compounds of C. sinensis, and the mycelium extract had significant antitumour activity. Significance and Impact of the Study:, The Cs-HK1 fungus may be a new and promising medicinal fungus and an effective and economical substitute of the wild C. sinensis for health care. [source] OPTIMAL CONDITIONS FOR THE GROWTH AND POLYSACCHARIDE PRODUCTION BY HYPSIZIGUS MARMOREUS IN SUBMERGED CULTUREJOURNAL OF FOOD PROCESSING AND PRESERVATION, Issue 4 2009PING WANG ABSTRACTS In submerged cultivation, many nutrient variables and environmental conditions have great influence on the growth and polysaccharide production by Hypsizigus marmoreus. Plackett,Burman design was used to determine the important nutrient factors. A central composite experimental design and surface response methodology were employed to optimize the factor levels. Prediction models for dry cell weight (DCW), polysaccharide outside cells (EPS) and polysaccharide inside cells (IPS) under important nutrient conditions were developed by multiple regression analysis and verified. By solving the equations, the optimal nutrient conditions for highest EPS production (9.62 g/L) were obtained at 6.77 g cornstarch/L, 36.57 g glucose/L, 3.5 g MgSO4/L and 6.14 g bean cake powder/L, under which DCW and IPS were 16.2 g/L and 1.46 g/L, close to the highest value under their corresponding optimal conditions. Optimal environmental conditions were obtained at 10% inoculation dose, 45 mL medium in a 250 mL flask, pH 6.5, 25C and 200 rpm according to the results of single-factor experiment design. PRACTICAL APPLICATIONS Hypsizigus marmoreus polysaccharides have many functional properties, including antitumor, antifungal and antiproliferative activities, and free-radical scavenging. Liquid cultivation could produce a higher yield of polysaccharides and more flexible sequential processing methods of H. marmoreus, compared with traditional solid-state cultivation. However, the cell growth and production of polysaccharides would be influenced by many factors, including nutrient conditions and environmental conditions in the liquid cultivation of H. marmoreus. Keeping the conditions at optimal levels can maximize the yield of polysaccharides. The study not only found out the optimal nutrient conditions and environmental conditions for highest cell growth and yield of polysaccharides, but also developed prediction models for these parameters with important nutrient variables. Yield of polysaccharide inside of cells was also studied as well as polysaccharides outside of cells and cell growth. The results provide essential information for production of H. marmoreus polysaccharides by liquid culture. [source] Identification and Regulation of Genes from a Biocontrol Strain of Fusarium oxysporumJOURNAL OF PHYTOPATHOLOGY, Issue 9 2007D. R. Fravel Abstract Differential display with three time points revealed that thiram altered expression of numerous genes in the biocontrol fungus Fusarium oxysporum CS-20. Of the 101 bands purified from the differential display gel, 86 were successfully cloned, and 64 sequenced. Based on nucleic acid sequences, homology to known products was found using BLASTn for 26 sequences and homology to hypothetical proteins was found for six sequences, also from Gibberella zeae. One band (BM1 24-1) showed homology to an ABC transporter from three different fungi. Because of its association with detoxification functions, the ABC transporter was selected for further study. Mycelia of CS-20 were exposed to 25 ,g active ingredient (a.i.) thiram in liquid culture for various times from 0 to 8 h. Quantitative real-time PCR was used to evaluate gene expression. At 30 min after treatment with thiram, the ABC transporter was upregulated 20- to 25-fold relative to the control treatment. The ABC transporter was upregulated 15-fold at 1 h after treatment and 10-fold at 2 h. At 8 h after treatment, there was no difference between treated and non-treated for expression of the ABC transporter. Transcription of the gene encoding EST BM1 24-1 is induced in response to thiram treatment and may function in providing resistance in F. oxysporum isolate CS-20 to fungicides and other toxins. Tolerance to toxins may be critical to the successful inclusion of CS-20 in disease control strategies in cropping systems. [source] Chromatographic Characterization and Phytotoxic Activity of Fusarium oxysporum f. sp. albedinis and Saprophytic Strain ToxinsJOURNAL OF PHYTOPATHOLOGY, Issue 4 2005H. Amraoui Abstract The bayoud disease, vascular fusariosis of date palm tree (Phoenix dactylifera L.), is caused by the pathogenic fungus Fusarium oxysporum f. sp. albedinis. The characteristic symptoms of the bayoud disease were elicited on detached leaves of F. oxysporum f. sp. albedinis -susceptible cultivars of date palm trees, which were treated either with the FII (F. oxysporum f. sp. albedinis) fraction purified from the organic extracts of a F. oxysporum f. sp. albedinis liquid culture, or with a solution of fusaric acid. Enniatins, which are secreted by several Fusarium species, were tested at different concentrations and were not capable of inducing symptoms on such detached leaves. The FII (F. oxysporum f. sp. albedinis) fraction was unable to induce necrosis of potato slices, which indicates that it does not contain significant amounts of enniatins. The high-performance liquid chromatography (HPLC) profiles of the FII (F. oxysporum f. sp. albedinis) fraction showed toxic peaks different from fusaric acid. A fraction, named FII (AZ4), was obtained from culture filtrates of a saprophytic Fusarium strain maintained in the same cultural conditions as for the F. oxysporum f. sp. albedinis. The HPLC profile of the FII (AZ4) fraction did not show the characteristic phytotoxic peaks present in the FII (F. oxysporum f. sp. albedinis) fraction. This finding well agrees with the fact that the FII (AZ4) fraction is not toxic to detached date palm leaves. Moreover, the HPLC profiles of FII fractions obtained from other special forms of F. oxysporum are different the FII (F. oxysporum f. sp. albedinis) profile. The phytotoxic compounds purified from the FII (F. oxysporum f. sp. albedinis) fraction are probably new molecules that may help in understanding the pathogenesis of bayoud disease. [source] Streptococcus pyogenes pili promote pharyngeal cell adhesion and biofilm formationMOLECULAR MICROBIOLOGY, Issue 4 2007Andrea G. O. Manetti Summary Group A Streptococcus (GAS, Streptococcus pyogenes) is a Gram-positive human pathogen responsible for several acute diseases and autoimmune sequelae that account for half a million deaths worldwide every year. GAS infections require the capacity of the pathogen to adhere to host tissues and assemble in cell aggregates. Furthermore, a role for biofilms in GAS pathogenesis has recently been proposed. Here we investigated the role of GAS pili in biofilm formation. We demonstrated that GAS pilus-negative mutants, in which the genes encoding either the pilus backbone structural protein or the sortase C1 have been deleted, showed an impaired capacity to attach to a pharyngeal cell line. The same mutants were much less efficient in forming cellular aggregates in liquid culture and microcolonies on human cells. Furthermore, mutant strains were incapable of producing the typical three-dimensional layer with bacterial microcolonies embedded in a carbohydrate polymeric matrix. Complemented mutants had an adhesion and aggregation phenotype similar to the wild-type strain. Finally, in vivo expression of pili was indirectly confirmed by demonstrating that most of the sera from human patients affected by GAS-mediated pharyngitis recognized recombinant pili proteins. These data support the role of pili in GAS adherence and colonization and suggest a general role of pili in all pathogenic streptococci. [source] Small genetic differences between ericoid mycorrhizal fungi affect nitrogen uptake by VacciniumNEW PHYTOLOGIST, Issue 3 2009Gwen-Aëlle Grelet Summary ,,Ericoid mycorrhizal fungi have been shown to differ in their pattern of nitrogen (N) use in pure culture. Here, we investigate whether this functional variation is maintained in symbiosis using three ascomycetes from a clade not previously shown to include ericoid mycorrhizal taxa. ,,Vaccinium macrocarpon and Vaccinium vitis-idaea were inoculated with three fungal strains known to form coils in Vaccinium roots, which differed in their patterns of N use in liquid culture. 15N was used to trace the uptake of -N, -N and glutamine-N into shoots. ,,15N transfer differed among the three fungal strains, including two that had identical internal transcribed spacer (ITS) sequences, and was quantitatively related to fungal growth in liquid culture at low carbon availability. ,,These results demonstrate that functional differences among closely related ericoid mycorrhizal fungi are maintained in symbiosis with their hosts, and suggest that N transfer to plant shoots in ericoid mycorrhizas is under fungal control. [source] The sugar porter gene family of Laccaria bicolor: function in ectomycorrhizal symbiosis and soil-growing hyphaeNEW PHYTOLOGIST, Issue 2 2008Mónica Fajardo López Summary ,,Formation of ectomycorrhizas, a symbiosis with fine roots of woody plants, is one way for soil fungi to overcome carbohydrate limitation in forest ecosystems. ,,Fifteen potential hexose transporter proteins, of which 10 group within three clusters, are encoded in the genome of the ectomycorrhizal model fungus Laccaria bicolor. For 14 of them, transcripts were detectable. ,,When grown in liquid culture, carbon starvation resulted in at least twofold higher transcript abundances for seven genes. Temporarily elevated transcript abundance after sugar addition was observed for three genes. Compared with the extraradical mycelium, ectomycorrhiza formation resulted in a strongly enhanced expression of six genes, of which four revealed their highest observed transcript abundances in symbiosis. A function as hexose importer was proven for three of them. Only three genes, of which just one was expressed at a considerable level, revealed a reduced transcript content in mycorrhizas. ,,From gene expression patterns and import kinetics, the L. bicolor hexose transporters could be divided into two groups: those responsible for uptake of carbohydrates by soil-growing hyphae, for improved carbon nutrition, and to reduce nutrient uptake competition by other soil microorganisms; and those responsible for efficient hexose uptake at the plant,fungus interface. [source] Diversity and salt tolerance of native Acacia rhizobia isolated from saline and non-saline soilsAUSTRAL ECOLOGY, Issue 8 2009PETER H. THRALL Abstract Re-establishing native vegetation in stressed soils is of considerable importance in many parts of the world, leading to significant interest in using plant,soil symbiont interactions to increase the cost-effectiveness of large-scale restoration. However, effective use of soil microbes in revegetation requires knowledge of how microbe communities vary along environmental stress gradients, as well as how such variation relates to symbiont effectiveness. In Australia, shrubby legumes dominate many ecosystems where dryland salinity is a major issue, and improving plant establishment in saline soils is a priority of regional management agencies. In this study, strains of rhizobial bacteria were isolated from a range of Acacia spp. growing in saline and non-saline soils. Replicates of each strain were grown under several salinity levels in liquid culture and characterized for growth and salt tolerance. Genetic characterization of rhizobia showed considerable variation among strains, with salt tolerance and growth generally higher in rhizobial populations derived from more saline soils. These strains showed markedly different genetic profiles and generic affiliations to those from more temperate soils, suggesting community differentiation in relation to salt stress. The identification of novel genomic species from saline soils suggests that the diversity of rhizobia associated with Australian Acacia spp. is significantly greater than previously described. Overall, the ability of some symbiotically effective strains to tolerate high salinity is promising with regard to improving host plant re-establishment in these soils. [source] Simultaneous Analysis of Spatio-Temporal Gene Expression for Cephamycin Biosynthesis in StreptomycesclavuligerusBIOTECHNOLOGY PROGRESS, Issue 6 2001Yun Seung Kyung Linkage between structural and regulatory genes implies that a direct correlation should exist between the spatio-temporal distribution of their expression. Green fluorescent protein (GFP) and cyan fluorescent protein (CFP) were used as reporters to analyze simultaneously expression of lysine-,-aminotransferase (LAT) and its corresponding genetic regulator, CcaR. The isogenic strain containing lat::gfp and ccaR::cfp in the chromosome produced cephamycin C at levels similar to wild type Streptomyces clavuligerus. Confocal laser scanning microscopy revealed that expression of both LAT and CcaR in liquid culture was temporally dynamic and spatially heterogeneous in S.clavuligerus mycelia. During the early culture stage only a part of the mycelia began to express LAT and CcaR at low levels. As the culture aged, expression levels and the population of mycelia expressing LAT and CcaR increased and were followed late in the growth cycle by a reduction of the mycelia population expressing LAT and CcaR. The approach provides a precise simultaneous temporal-spatial expression profile and corroborates the regulatory linkage between ccaR and lat in S. clavuligerus. [source] Thrombopoietin, flt3-ligand and c-kit-ligand modulate HOX gene expression in expanding cord blood CD133+ cellsCELL PROLIFERATION, Issue 4 2004C. P. McGuckin Extrinsic modulators include growth factors and cell adhesion molecules, whereas intrinsic regulation is achieved with many transcription factor families, of which the HOX gene products are known to be important in haemopoiesis. Umbilical cord blood CD133+ HSPC proliferation potential was tested in liquid culture with ,TPOFLK' (thrombopoietin, flt-3 ligand and c-kit ligand, promoting HSPC survival and self-renewal), in comparison to ,K36EG' (c-kit-ligand, interleukins-3 and -6, erythropoietin and granulocyte colony-stimulating factor, inducing haemopoietic differentiation). TPOFLK induced a higher CD133+ HSPC proliferation (up to 60-fold more, at week 8) and maintained a higher frequency of the primitive colony-forming cells than K36EG. Quantitative polymerase chain reaction analysis revealed opposite expression patterns for specific HOX genes in expanding cord blood CD133+ HSPC. After 8 weeks in liquid culture, TPOFLK increased the expression of HOX B3, B4 and A9 (associated with uncommitted HSPC) and reduced the expression of HOX B8 and A10 (expressed in committed myeloid cells) when compared to K36EG. These results suggest that TPOFLK induces CD133+ HSPC proliferation, self-renewal and maintenance, up-regulation of HOX B3, B4 and A9 and down-regulation of HOX B8 and A10 gene expression. [source] Volatiles Released by a Streptomyces Species Isolated from the North SeaCHEMISTRY & BIODIVERSITY, Issue 7 2005Jeroen The North Sea Streptomyces strain GWS-BW-H5 was investigated by analyzing headspace extracts of agar-plate cultures (HE) or liquid cultures (LCE), obtained with a closed-loop stripping apparatus (CLSA), by GC/MS (Table,1). The volatile profile of the HE is dominated by the known volatiles (,)-geosmin (4) and 2-methyisoborneol (1). Small amounts of sesquiterpenes occur, which are present in a more-diverse structural variety and in higher quantities in the LCE. The different structures can be rationalized by few cationic intermediates along their biosynthetic pathway. The most-prominent difference between the two culture methods were the presence of the Me-branched , - and , -lactones 31,38, not previously reported from nature, in the LCE. Major components were 10-methyldodecan-5-olide (34), 10-methyldodec-2-en-4-olide (36), and 10-methyldodec-3-en-4-olide (38). The structures of all new lactones were verified by synthesis. Furthermore, more volatiles in higher amounts were produced by the liquid culture than by to the agar plate culture. Since 36 showed inhibitory growth effects against strain GWS-BW-H5, growth inhibition against twelve other strains isolated from the same habitat was tested. Antagonistic activity against four of the strains was observed, with a slightly higher threshold level than found for penicillin G, which was used in control experiments (Table,2). [source] Supernatants of Pseudomonas aeruginosa induce the Pseudomonas -specific antibiotic elafin in human keratinocytesEXPERIMENTAL DERMATOLOGY, Issue 4 2003Ulf Meyer-Hoffert Abstract: Elafin is a skin-derived serine-protease inhibitor. It is thought to be important to prevent human leukocyte elastase-mediated tissue damage and might play an important role in maintaining the integrity of the human epidermis. Recent studies have provided evidence for an antimicrobial activity of elafin against P. aeruginosa. As gram-negative infections typically occur in barrier-disrupted skin we were interested to determine whether supernatants of the gram-negative bacteria P. aeruginosa and Escherichia coli were capable of inducing elafin expression. Supernatants of various P. aeruginosa strains stimulated elafin mRNA-expression and protein release, whereas supernatants of E. coli did not induce elafin expression. In non-differentiated cells the relative increase of elafin mRNA was much higher (100-fold) than in differentiated cells (sixfold), although the latter exhibited higher constitutive mRNA-expression (150-fold). However, concentrations of secreted elafin were similar in differentiated and non-differentiated cells after stimulation. We could not confirm a bactericidal effect against P. aeruginosa as described previously but observed that its growth was inhibited as demonstrated for different strains in liquid cultures. Growth of E. coli was not affected by elafin. In conclusion, the data presented in this paper suggest that elafin represents an innate immune response factor induced by secreted products of P. aeruginosa. Besides its elastase inhibitory potency elafin is an antimicrobial agent against P. aeruginosa. [source] Lead and cadmium uptake in the marine fungi Corollospora lacera and Monodictys pelagicaFEMS MICROBIOLOGY ECOLOGY, Issue 3 2005Michael A.S. Taboski Abstract This study provides observations on the effects of lead and cadmium ions on the growth of two species of marine fungi, Corollospora lacera and Monodictys pelagica. On solid media lead appeared to have no effect on the radial rate of growth of fungi. Exposure to increasing cadmium concentrations on solid media resulted in significant reduction (P < 0.05) in the radial mycelial growth rates of both fungi, especially in M. pelagica. These results reveal significant difference in species sensitivity toward cadmium and, essentially, insensitivity toward lead exposure. In liquid cultures, the metal content of mycelia (metal mass found in mycelium, in mg), and the concentration of metal in dry mycelium (metal mass in 1 g of mycelium, in mg g,1) were both found to increase (P < 0.05) with the increase in the metal cation concentration, while mycelium dry mass decreased. As it was observed on solid media, cadmium cation affected more severely (P < 0.05) the growth of M. pelagica in liquid cultures. Ergosterol content of mycelia of C. lacera exposed to increasing cadmium cation concentration decreased, similarly to the trend observed for dry mycelial mass. It was found that ca. 93% of all lead sequestered by C. lacera is located extracellularly. M. pelagica was found to bioaccumulate over 60 mg of cadmium and over 6 mg of lead per 1 g of mycelium, while C. lacera bioaccumulated over 7 mg of cadmium and up to 250 mg of lead per 1 g of mycelium. Overall, the results indicate that both metal ions affect the growth of marine fungi with lead being accumulated extracellularly in the mycelia. Both metals accumulated by fungi may then enter the marine ecosystem food web, of which marine fungi are integral members. [source] Accumulation of deoxynivalenol and its 15-acetylated form is significantly modulated by oxidative stress in liquid cultures of Fusarium graminearumFEMS MICROBIOLOGY LETTERS, Issue 1 2006Nadia Ponts Abstract Liquid cultures of Fusarium graminearum were supplemented with H2O2 or other oxidative compounds. The accumulation kinetics of the resulting trichothecenes were monitored. At non-lethal concentrations, the H2O2 treatments modulated toxin accumulation, dependent on the method of supplementation. When H2O2 was added at the same time as the inoculation, higher levels of toxins accumulated 30 days later. Conversely, adding H2O2 2 or 7 days after inoculation had little effect. When H2O2 was added daily over the course of the culture, the accumulation of trichothecenes was rapidly and strongly enhanced. The fungus may adapt to oxidative stress when the first exposure to H2O2 occurs at the beginning of the culture course. The highest toxin levels were measured when the H2O2 was added daily. The importance of the first hours of culture was confirmed: pre-treating conidia with H2O2 does not affect their germination kinetics but leads to a reduction in the yield of trichothecenes 40 days later. The H2O2 regulation of this trichothecene accumulation may be specific, as paraquat, another pro-oxidant compound, inhibits their production. Since H2O2 is a major component of the oxidative burst occurring in pathogen/host interactions, these data support the theory that trichothecenes may act as virulence factors. [source] Is Helicobacter pylori a True Microaerophile?HELICOBACTER, Issue 4 2006Stephanie Bury-Moné Abstract Background:, There is no general consensus about the specific oxygen and carbon dioxide requirements of the human pathogen Helicobacter pylori. This bacterium is considered a microaerophile and consequently, it is grown under atmospheres at oxygen tensions 5,19% and carbon dioxide tensions 5,10%, both for clinical and basic and applied research purposes. The current study compared the growth of H. pylori in vitro, under various gas atmospheres, and determined some specific changes in the physiology of bacteria grown under different oxygen partial pressures. Methods:, Measurements of bacterial growth under various conditions were carried out employing classical solid and liquid culture techniques. Enzymatic activities were measured using spectrophotometric assays. Results:,H. pylori and all the other Helicobacter spp. tested had an absolute requirement for elevated carbon dioxide concentrations in the growth atmosphere. In contrast with other Helicobacter spp., H. pylori can tolerate elevated oxygen tensions when grown at high bacterial concentrations. Under 5% CO2, the bacterium showed similar growth in liquid cultures under oxygen tensions from microaerobic (< 5%) to fully aerobic (21%) at cell densities higher than 5 × 105 cfu/ml for media supplemented with horse serum and 5 × 107 cfu/ml for media supplemented with ,-cyclodextrin. Evidence that changes occurred in the physiology of H. pylori was obtained by comparing the activities of ferredoxin:NADH (nicotinamide adenine dinucleotide) oxidoreductases of bacteria grown under microaerobic and aerobic atmospheres. Conclusions:,H. pylori is a capnophile able to grow equally well in vitro under microaerobic or aerobic conditions at high bacterial concentrations, and behaved like oxygen-sensitive microaerophiles at low cell densities. Some characteristics of H. pylori cells grown in vitro under microaerobic conditions appeared to mimic better the physiology of organisms grown in their natural niche in the human stomach. [source] Relationship Between Gastric Ulcer and Helicobacter pylori VacA Detected in Gastric Juice Using Bead-ELISA MethodHELICOBACTER, Issue 5 2002Daisuke Shirasaka Abstract Background. VacA is an important pathogenetic factor produced by Helicobacter pylori. VacA has often been detected in supernatants of liquid cultures or lysates of whole bacterial cells. However, no studies have ever tried to assay VacA produced in the human stomach. We applied a very sensitive and simple method, bead-ELISA, to detect VacA in gastric juice. Materials and Methods. Forty-eight H. pylori -positive patients (16 nonulcer dyspepsia, 16 gastric ulcer, and 16 duodenal ulcer) and four H. pylori -negative nonulcer dyspepsia patients had endoscopy performed and gastric juice were aspirated. Polystyrene beads coated with the antibody to VacA, were used in this bead-ELISA method. The nucleotide sequences of vacA in the signal and middle regions were investigated. Results. Of the 48 samples that were positive for H. pylori, 21 [43.8%] were found to be VacA positive in gastric juice. The average and maximum concentrations of detected VacA in gastric juice were 143.2 ± 216.5 and 840 pg/ml, respectively. The average density of VacA from gastric ulcer patients (227.5 ± 276.7 pg/ml) was higher than that found in nonulcer dyspepsia (51.8 ± 39.8 pg/ml) and duodenal ulcer (49.2 ± 21.5 pg/ml) patients. There was no relationship between VacA in gastric juice and vacA genotype. Conclusions. VacA in gastric juice could be directly detected by bead-ELISA. In this study, the diversity of disease outcome was associated with not the quality but the quantity of VacA. Therefore, not only the quality but also the quantity of VacA is important etiological factors in the pathogenesis of mucosal damage. [source] Effect of different liquid cultures of live yeast strains on performance, ruminal fermentation and microbial protein synthesis in lambsJOURNAL OF ANIMAL PHYSIOLOGY AND NUTRITION, Issue 6 2008M. K. Tripathi Summary Three yeast strains, Kluyveromyces marximanus NRRL-3234 (KM), Saccharomyces cerevisiae NCDC-42 (SC) and Saccharomyces uvarum ATCC-9080 (SU), and a mixed culture (1:1:1 ratio) were evaluated for their value as probiotics in lamb feeding in two experiment. In experiment I and II, 20 and 30 pre-weaner lambs were fed for 63 and 60 days in two and three equal groups respectively. All lambs were offered ad libitum a creep mixture and Zizyphus nummularia leaves, and yeasts were dosed orally. In experiment I, one group received no yeast, the other of the mixed culture (1.5,2 × 1010 live cells/ml). In experiment II, yeast cultivation was modified yielding 1.5,2 × 1013 live cells/ml. Lambs of the three experimental groups received 1 ml/kg live weight of one of the individual yeasts. Feed intake did not differ among groups of both experiments with the exception of SC-supplemented lambs in experiment II which showed a trend to higher intakes per kg metabolic body weight and in percentage of body weight when compared with KM- and SU-supplemented lambs. Supplementation of the mixed yeast culture had no effect on intakes of digestible crude protein and metabolisable energy, nutrient digestibility, nitrogen balance and rumen fermentation characteristics (pH, ammonia, volatile fatty acid concentration, protozoa count) and urinary allantoin as an indicator of microbial protein synthesis. The same was true for comparisons in experiment II except ciliate protozoa counts, which showed a trend to be the highest with SU and the lowest with SC. The results of present study show that the response of lambs to supplemented live yeast cultures is inconsistent, as it lacked to have an effect in the present study, and that differences among strains were small, even when supplemented at a much higher live cell count. [source] Overexpression of hns in the plant growth-promoting bacterium Enterobacter cloacae UW5 increases root colonizationJOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2010M.M. English Abstract Aims:, Plant growth-promoting rhizobacteria (PGPR) introduced into soil often do not compete effectively with indigenous micro-organisms for plant colonization. The aim of this study was to identify novel genes that are important for root colonization by the PGPR Enterobacter cloacae UW5. Methods and Results:, A library of transposon mutants of Ent. cloacae UW5 was screened for mutants with altered ability to colonize canola roots using a thermal asymmetric interlaced (TAIL)-PCR-based approach. A PCR fragment from one mutant was reproducibly amplified at greater levels from genomic DNA extracted from mutant pools recovered from seedling roots 6 days after seed inoculation compared to that from the cognate inoculum cultures. Competition assays confirmed that the purified mutant designated Ent. cloacae J28 outcompetes the wild-type strain on roots but not in liquid cultures. In Ent. cloacae J28, the transposon is inserted upstream of the hns gene. Quantitative RT-PCR showed that transposon insertion increased expression of hns on roots. Conclusions:, These results indicate that increased expression of hns in Ent. cloacae enhances competitive colonization of roots. Significance and Impact of the Study:, A better understanding of the genes involved in plant colonization will contribute to the development of PGPR that can compete more effectively in agricultural soils. [source] Sequential secretion of collagenolytic, elastolytic, and keratinolytic proteases in peptide-limited cultures of two Bacillus cereus strains isolated from woolJOURNAL OF APPLIED MICROBIOLOGY, Issue 1 2009A.C. Ad, güzel Abstract Aims:, To characterize the secretion of proteolytic activities against keratin, collagen and elastin in liquid cultures of Bacillus cereus IZ-06b and IZ-06r isolated from wool. Methods and Results:, Growth of B. cereus IZ-06b and IZ-06r were characterized in batch culture. Both strains needed an organic nitrogen source, were able to grow on wool or peptone as sole carbon and nitrogen sources, and metabolized glucose, maltose and other simple sugars. Proteolytic activities were investigated in batch cultures grown in peptide-restricted, carbon-sufficient medium. Secretion of proteases was induced by peptide limitation while different proteolytic activities appeared sequentially in the growth medium. When the most available components of the peptone were depleted, collagenolytic and elastolytic proteases were produced. These were later replaced by the production of keratinolytic protease. Conclusions:,B. cereus can adjust its proteolytic affinity profile in response to the supply of organic nitrogen and sequentially secrete proteases with activities targeted against increasingly inaccessible proteinous substrates as the nutritional availability in the environment deteriorates. Significance and Impact of the Study:, Peptide-limited, carbon-sufficient growth media containing no proteinous substrates are well suited for protease production in B. cereus while growth conditions can be adjusted to optimize the proteolytic affinity profiles. [source] Mycelium cultivation, chemical composition and antitumour activity of a Tolypocladium sp. fungus isolated from wild Cordyceps sinensisJOURNAL OF APPLIED MICROBIOLOGY, Issue 2 2006P.H. Leung Abstract Aims:, To examine and illustrate the morphological characteristics and growth kinetics of Cs-HK1, a Tolypocladium fungus, isolated from wild Cordyceps sinensis in solid and liquid cultures, and the major chemical constituents and antitumour effects of Cs-HK1 mycelium. Methods and Results:, The Cs-HK1 fungus was isolated from the fruiting body of a wild C. sinensis and identified as a Tolypocladium sp. fungus. It grew rapidly at 22,25°C on a liquid medium containing glucose, yeast extract, peptone and major inorganic salts, with a specific growth rate of 1·1 day,1, reaching a cell density of 23·0 g dw l,1 in 7,9 days. Exopolysaccharides accumulated in the liquid culture to about 0·3 g l,1 glucose equivalent. In comparison with natural C. sinensis, the fungal mycelium had similar contents of protein (11·7,,g) and carbohydrate (654·6,,g) but much higher contents of polysaccharide (244·2 mg vs 129·5 mg), adenosine (1116·8,,g vs 264·6 ,g) and cordycepin (65·7 ,g vs 20·8 ,g) (per gram dry weight). Cyclosporin A, an antibiotic commonly produced by Tolypocladium sp., was also detected from the mycelium extract. The hot water extract of mycelium showed low cytotoxic effect on B16 melanoma cells in culture (about 25% inhibition) but significant antitumour effect in animal tests, causing 50% inhibition of B16 cell-induced tumour growth in mice. Conclusions:, The Tolypocladium sp. fungus, Cs-HK1, can be easily cultivated by liquid fermentation. The mycelium biomass contained the major bioactive compounds of C. sinensis, and the mycelium extract had significant antitumour activity. Significance and Impact of the Study:, The Cs-HK1 fungus may be a new and promising medicinal fungus and an effective and economical substitute of the wild C. sinensis for health care. [source] 97 Sensitivity of cyanobacteria to a potential biological control agent, bacterium SG-3JOURNAL OF PHYCOLOGY, Issue 2003K. Wilkinson Cyanobacteria cause many problems in freshwater ecosystems. For example, the production of off-flavor compounds by cyanobacteria causes serious problems in catfish aquaculture. Control of cyanobacteria is generally limited to treatment with copper compounds, which are non-selective and sometimes ineffective at controlling certain species of cyanobacteria. Biological control could provide selective management by removing unwanted species while leaving desirable algae species. A bacterium (SG-3) (NRRL B-30043) lyses a number of planktonic species of cyanobacteria including bloom-forming species of Anabaena and Oscillatoria. We tested SG-3 for activity against 10 isolates, representing seven species, of mat-forming cyanobacteria within the genera Oscillatoria, Lyngbya, and Phormidium. Plugs (0.5 cm diameter) were cut from mats of the cyanobacterium, inoculated with liquid cultures of SG-3, and incubated as static cultures. The reduction in dry weights ranged from ,0.5% to 90% compared to the untreated controls and appeared to be species specific. For example, dry weight reductions of Oscillatoria deflexoides and O. amoena ranged from 80 to 90% whereas the reduction of O. limosa tended to be lower at 36 to 72%. Although results varied among and within species, they indicate that this bacterium could have potential for use as a biological control for mat-forming cyanobacteria. Light microscopic observations indicate the bacteria do not penetrate the cyanobacteria cells. Currently, we are studying the possible causes of the observed cell lysis. [source] Solvent extraction of bacteriocins from liquid culturesLETTERS IN APPLIED MICROBIOLOGY, Issue 3 2000L.L. Burianek A solvent extraction method was developed to concentrate lacidin from the culture of Lactobacillus acidophilus OSU133. The new method concentrates the bacteriocin at the interface between chloroform and the aqueous culture of the producing bacterium. Compared with other extraction procedures, the new method effectively recovers higher bacteriocin yield and results in relatively clean preparations. Recovery of lacidin by the chloroform extraction procedure, compared with ammonium sulphate precipitation and cell acidification methods, was >10-fold and about 100-fold greater, respectively. The new extraction procedure saves time and is easy to perform. This method is also effective in recovering subtilin, bacillicin, pediocin and nisin from cultures of Bacillus subtilis ATCC 6633, B. subtilis OSY1115/C, Pediococcus acidilactici PO2 and Lactococcus lactis ATCC 11454, respectively. [source] Comparison of endo-polygalacturonase activities of citrus and non-citrus races of Geotrichum candidum, and cloning and expression of the corresponding genesMOLECULAR PLANT PATHOLOGY, Issue 5 2001Masayuki Nakamura summary Geotrichum candidum citrus race, a fungus that causes a sour rot disease in citrus fruits, secretes an endo-polygalacturonase (PG) that may facilitate the disease. There also exists a non-citrus race that is non-pathogenic to citrus fruits. In this research, we found that the PG activity of the citrus race isolates was much higher than that of the non-citrus race isolates in culture medium and inoculated lemon peel, and that there was a significant correlation between the PG activity and pathogenicity. We isolated the two corresponding PG genes, S31pg1 and S63pg1, from citrus race S31 and non-citrus race S63, respectively. S31PG1 and S63PG1 consisted of 368 and 369 amino acids, respectively. The two PG genes showed 68% identity at the amino acid level. In expression studies, S31pg1 transcript was detected in mycelia grown in liquid cultures of citrus race S31 containing either glucose, pectin or lemon peel broth. The transcript was also detected in lemon peel inoculated with the isolate. On the other hand, no transcript of S63pg1 was detected in mycelia grown on any liquid cultures of non-citrus race S63 and lemon peel inoculated with the isolate. These results indicate that PG may play an important role in the development of the sour rot symptom and be involved in the difference of pathogenicity between the two races. [source] Mechanisms involved in control of Blumeria graminis f.sp. hordei in barley treated with mycelial extracts from cultured fungiPLANT PATHOLOGY, Issue 5 2002H. Haugaard Treatment with mycelial extracts, prepared from liquid cultures of Bipolaris oryzae, Pythium ultimum and Rhizopus stolonifer, protected barley (Hordeum vulgare) against powdery mildew disease caused by the fungus Blumeria graminis f.sp. hordei. The mechanisms of this protection were studied using histopathological methods and molecular analysis. Germination and appressorial formation of B. graminis were generally reduced after treatment with mycelial extracts. Although this reduction (between 12 and 62% depending on treatment and experiment) was inconsistent and only occasionally significantly different from the water-treated control, it indicated a direct antifungal effect of the extracts. In situations where the fungus succeeded in forming an appressorium, penetration efficiency and haustorium formation from these appressoria was not affected , no enhanced penetration resistance associated with papilla formation was detected. However, a post-penetration effect was observed, as B. graminis colonies on mycelial extract-treated leaves produced 50% fewer hyphae than on controls. Northern blot analyses showed earlier accumulation of defence-related gene transcripts following treatment with B. oryzae and P. ultimum mycelial extracts, and to a lesser extent R. stolonifer mycelial extract, compared with water-treated leaves. It is suggested that the protection mechanism of the mycelial extracts involves direct antifungal effects and possible induced resistance for the B. oryzae and P. ultimum mycelial extracts. [source] In situ localisation and quantification of surfactins in a Bacillus subtilis swarming community by imaging mass spectrometryPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 18 2008Delphine Debois Abstract Surfactins are a family of heptacyclopeptides in which the C-terminal carbonyl is linked with the ,-hydroxy group of a fatty acid acylating the N-terminal function of a glutamic acid residue. The fatty acyl chain is 12,16 carbon atoms long. These compounds, which are secreted by the Gram-positive bacterium Bacillus subtilis in stationary phase in liquid cultures, play an important role in swarming communities on the surface of agar media in the formation of dendritic patterns. TOF secondary ion MS (TOF-SIMS) imaging was used to map surfactins within 16,17,h swarming patterns, with a 2,,m spatial resolution. Surfactins were mainly located in the central mother colony (the site of initial inoculation), in a ,ring' surrounding the pattern and along the edges of the dendrites. In the mother colony and the interior of the dendrites, surfactins with shorter chain lengths are present, whereas in the ring surrounding the swarm community and between dendrites, surfactins with longer fatty acyl chain lengths were found. A quantitative analysis by MALDI-TOF MS showed a concentration gradient of surfactin from the mother colony to the periphery. The concentration of surfactin was ,400,pmol/mL in the mother colony and ,10,pmol/mL at the base of the dendrites, decreasing to 2,pmol/mL at their tips. [source] Time-series integrated "omic" analyses to elucidate short-term stress-induced responses in plant liquid cultures,BIOTECHNOLOGY & BIOENGINEERING, Issue 1 2009Bhaskar Dutta Abstract The research that aims at furthering our understanding of plant primary metabolism has intensified during the last decade. The presented study validated a systems biology methodological framework for the analysis of stress-induced molecular interaction networks in the context of plant primary metabolism, as these are expressed during the first hours of the stress treatment. The framework involves the application of time-series integrated full-genome transcriptomic and polar metabolomic analyses on plant liquid cultures. The latter were selected as the model system for this type of analysis, because they provide a well-controlled growth environment, ensuring that the observed plant response is due only to the applied perturbation. An enhanced gas chromatography,mass spectrometry (GC,MS) metabolomic data correction strategy and a new algorithm for the significance analysis of time-series "omic" data are used to extract information about the plant's transcriptional and metabolic response to the applied stress from the acquired datasets; in this article, it is the first time that these are applied for the analysis of a large biological dataset from a complex eukaryotic system. The case-study involved Arabidopsis thaliana liquid cultures subjected for 30 h to elevated (1%) CO2 stress. The advantages and validity of the methodological framework are discussed in the context of the known A. thaliana or plant, in general, physiology under the particular stress. Of note, the ability of the methodology to capture dynamic aspects of the observed molecular response allowed for 9 and 24 h of treatment to be indicated as corresponding to shifts in both the transcriptional and metabolic activity; analysis of the pathways through which these activity changes are manifested provides insight to regulatory processes. Biotechnol. Bioeng. 2009;102: 264,279. © 2008 Wiley Periodicals, Inc. [source] | |||||||||||||||||||||||||||||||||||||