bluuebunny/crossref_metadata_2025 huggingface dataset:
Double-stranded RNA mycoviruses in species ofAspergillussectionsCircumdatiandFumigati
Authors: Varga János , Rinyu Edit , Kevei Éva , Tóth Beáta , Kozakiewicz Zofia
Published: April 2011

Isolates (178) belonging to Aspergillus sections Fumigati, Candidi, Clavati, and Circumdati were tested for the presence of double-stranded RNA (dsRNA) genomes. Altogether, 5.6% of the Aspergillus strains examined were infected with dsRNAs. dsRNA segments indicative of mycovirus infection were observed for the first time in Neosartorya hiratsukae, Neosartorya quadricincta, Petromyces alliaceus, and Aspergillus clavatus strains. Correlation was not observed between ochratoxin production and dsRNA content of the strains. This is the first report on the detection of naturally occurring dsRNAs in Aspergillus species that are able to reproduce sexually. The detection of dsRNA in sexual aspergilli gave us a chance to examine the transmission of these segments through ascospores. A Neosartorya hiratsukae strain transmitted the dsRNAs efficiently through sexual spores, while the stromata embedding the asci in Petromyces alliaceus did not transmit one of the dsRNA segments. The 0.6-kb dsRNA segment that was present in the single-stromatal cultures was found to be located in the mitochondrial fraction of this strain. This observation indicates that some mechanisms exist in aspergilli to exclude cytoplasmically located dsRNA molecules from stromatal structures.Key words: Aspergillus, double-stranded RNA, mycovirus, Petromyces, Neosartorya.
Identification of σ32-like factors and ftsX-rpoH gene arrangements in enteric bacteria
Authors: Ramírez-Santos Jesús , Gómez-Eichelmann M Carmen
Published: April 2011

Western blot analyses using anti-Escherichia coli K-12 σ32 antibodies and Southern blot analyses using rpoH and ftsX DNA probes were performed using different enteric bacteria. Results show that the bacterial strains analysed have σ32-like transcription factors and ftsX and rpoH homologs in a similar map position. Although the presence of σ32-like factors seems to be extended to all Proteobacteria, rpoH and ftsX homologs seem to be present as neighbors in the genome only in the enteric bacteria.Key words: enteric bacteria, heat shock, σ32, ftsX-rpoH.
Acidity and calcium interaction affecting cell envelope stability inRhizobium
Authors: Ballen Karen G , Graham Peter H , Jones Roger K , Bowers John H
Published: April 2011

Calcium improves the ability of many rhizobia to survive and persist in acid soils, but the mechanism responsible for this phenomenon has not been studied in detail. Here, we present data examining the combined effects of pH and calcium on the cell envelope of Rhizobium strains that differ in pH tolerance. The effect of pH and calcium on solute uptake was demonstrated by a change in the resistance to selected antimicrobial agents. When grown at pH 5.0, all strains exhibited fatty acid methyl ester profiles that were significantly different from those obtained using cells grown at pH 7.0. These differences included changes in the C16:C18 ratio and the percentage of 19:0 cyclopropane in the membrane. Both pH and calcium level had marked effects on Rhizobium etli UMR1632 lipopolysaccharide-banding patterns, but there was little evidence of a change in lipopolysaccharides with pH and calcium in Rhizobium tropici UMR1899. Both pH and calcium influenced expression of outer membrane proteins in all strains.Key words: Rhizobium, acidity, calcium, lipopolysaccharide, cell envelope, outer membrane protein.
Influence of chemical surfactants on the biodegradation of crude oil by a mixed bacterial culture
Authors: Van Hamme J D , Ward O P
Published: April 2011

The effects of surfactant physicochemical properties, such as the hydrophile-lipophile balance (HLB) and molecular structure, on the biodegradation of 2% w/v Bow River crude oil by a mixed-bacterial culture were examined. Viable counts increased 4.6-fold and total petroleum hydrocarbon (TPH) biodegradation increased 57% in the presence of Igepal CO-630, a nonylphenol ethoxylate (HLB 13, 0.625 g/L). Only the nonylphenol ethoxylate with an HLB value of 13 substantially enhanced biodegradation. The surfactants from other chemical classes with HLB values of 13 (0.625 g/L) had no effect or were inhibitory. TPH biodegradation enhancement by Igepal CO-630 occurred at concentrations above the critical micelle concentration. When the effect of surfactant on individual oil fractions was examined, the biodegradation enhancement for the saturate and aromatic fractions was the same. In all cases, biodegradation resulted in increased resin and asphaltene concentrations. Optimal surfactant concentrations for TPH biodegradation reduced resin and asphaltene formation. Chemical surfactants have the potential to improve crude oil biodegradation in complex microbial systems, and surfactant selection should consider factors such as molecular structure, HLB, and surfactant concentration.Key words: mixed culture, crude oil, surfactant, hydrophile-lipophile balance, biodegradation.
Pyruvate metabolism byAnaplasma marginalein cell-free culture
Authors: McHolland Linda E , Caldwell Daniel R
Published: April 2011

Partially purified Anaplasma marginale initial bodies were cultivated in a cell-free system in the presence of [3-14C]pyruvate for 24 or 48 h. Experiments showed that a significant portion of the pyruvate supplied to the cultures was incorporated into initial body components. Label incorporation was reduced by 72% in the presence of oxytetracycline. Fractionation and chromatography of the organisms revealed radioactive incorporation as alanine. This is the first report of de novo amino acid synthesis by A. marginale demonstrating that the rickettsia is capable of using pyruvate, an erythrocyte glycolytic product, in its metabolism.Key words: Anaplasma marginale, pyruvate metabolism, amino acid synthesis.
Phenol degradation by an enterobacterium: aKlebsiellastrain carries a TOL-like plasmid and a gene encoding a novel phenol hydroxylase
Authors: Heesche-Wagner Kerstin , Schwarz Thomas , Kaufmann Michael
Published: April 2011

Although phenol catabolism is described for many different microorganisms, there is no example for such a pathway in an enterobacterial strain. Here we characterize a Klebsiella oxytoca strain that grows on phenol as the only source of carbon and energy. As the key enzyme of phenol degradation, phenol hydroxylase was purified to apparent homogeneity. Compared with other phenol hydroxylases, the Klebsiella enzyme differs with respect to several properties: (i) SDS-PAGE and gel-filtration analysis of the purified protein revealed that the enzyme is a monomer with a molecular mass of 156 kDa; (ii) steady-state kinetic measurements resulted in a Kmvalue of 0.22 mM for phenol; and (iii) the enzyme is both dependent on NADPH/FAD and sensitive to EDTA. Further degradation of catechol, the reaction product of phenol hydroxylase, may occur via the effective meta-fission pathway often located on TOL or TOL-like plasmids. Such a plasmid was prepared from the Klebsiella strain and further characterized. The given data demonstrate that the isolated strain exhibits all characteristics of an efficient phenol-degrading microorganism.Key words: phenol metabolism, Klebsiella oxytoca, phenol hydroxylase, TOL plasmids.
Conversion factor between acetylene reduction and nitrogen fixation in free-living cyanobacteria from high arctic habitats
Authors: Liengen Turid
Published: April 2011

The conversion factor between acetylene reduction and15N incorporation in free-living cyanobacteria was determined in different high arctic habitats in the area of Ny-Ålesund (78.5°N, 11.6°E), Spitsbergen, in the summer of 1994. The experiments were carried out under constant conditions, 19°C and 200 µE·m-2·s-1. The nitrogen-fixation activities, measured as15N-incorporation, were in the range 4.01-6.54 mg N2fixed·gdw-1·day-1(dw, dry weight) in sheets of Nostoc commune and 778-1206 mg N2fixed·m-2·day-1in the cyanobacterial crusts. The acetylene reduction activities were in the range 0.72-1.91 mg ethylene produced·gdw-1·day-1of N. commune and 12.8-63.7 mg ethylene produced·m-2·day-1in the cyanobacterial crusts. The conversion factor of N. commune ranged from 0.11 to 0.48 for ethylene produced to nitrogen fixed, whereas the cyanobacterial crusts covering the soil surface gave conversion factors in the range 0.022-0.073 for ethylene produced to nitrogen fixed. An Anabaena sp., isolated from one of the habitats investigated, gave conversion factors near the theoretical factor of 4, when determined at 14.0 and 17.3°C. It was concluded that the acetylene reduction activity of free-living cyanobacteria in high arctic habitats results in underestimates of the real nitrogen-fixation activity in these environments.Key words: nitrogen fixation, acetylene reduction, conversion factor, cyanobacteria, Nostoc commune, high arctic.
Microbial community structure and function in the spermosphere as affected by soil and seed type
Authors: Buyer Jeffrey S , Roberts Daniel P , Russek-Cohen Estelle
Published: April 2011

Colonization of the spermosphere and rhizosphere by plant-beneficial bacteria is limited by competition with indigenous soil microbes for resources such as reduced carbon compounds. A study of the soil microbial community around germinating seeds was undertaken as a necessary first step in understanding the competition between the introduced plant-beneficial bacteria and the indigenous microbial community. Two soil types, Galestown gravely loamy sand soil, with pH 5.8 and 0.6% humic material, and Hatborough loamy sand soil, with pH 4.5 and 3.2% humic material, were compared. Five seed types, corn, cucumber, radish, soybean, and sunflower were used. Microbial communities were characterized functionally, using a substrate utilization assay, and structurally, using fatty acid methyl ester analysis, over a 96-h period after the onset of germination. Soil type affected the microbial communities far more than seed type. The communities in Hatborough soil had greater functional and structural changes compared with the control than Galestown soil. The communities changed more functionally than structurally, with increased substrate utilization compared with the soil controls.Key words: spermosphere, microbial community, Biolog, substrate utilization, fatty acid.
Growth properties of afolAnull mutant ofEscherichia coliK12
Authors: Herrington Muriel B , Chirwa Neema T
Published: April 2011

In Escherichia coli, dihydrofolate reductase is required for both the de novo synthesis of tetrahydrofolate and the recycling of dihydrofolate produced during the synthesis of thymidylate. The coding region of the dihydrofolate reductase gene, folA, was replaced with a kanamycin resistance determinant. Unlike earlier deletions, this mutation did not disrupt flanking genes. When the mutation was transferred into a wild-type strain and a thymidine- (thy) requiring strain, the resulting strains were viable but slow growing on rich medium. Both synthesized less folate than their parents, as judged by the incorporation of radioactive para-aminobenzoic acid. The derivative of the wild-type strain did not grow on any defined minimal media tested. In contrast, the derivative of the thy-requiring strain grew slowly on minimal medium with thy but exhibited auxotrophies on some combinations of supplements. These results suggest that when folates are limited, they can be distributed appropriately to folate-dependent biosynthetic reactions only under some conditions. Key words: dihydrofolate reductase, Escherichia coli, biosynthesis, folates, one-carbon metabolism.
Survival of insect pathogenic and human clinical isolates ofPhotorhabdus luminescensin previously sterile soil
Authors: Bleakley Bruce H , Chen Xiang
Published: April 2011

Most characterized strains of the bacterium Photorhabdus luminescens are symbionts of entomopathogenic nematodes, whereas other strains have been isolated from human clinical specimens. The ability of P. luminescens strains to survive and grow in soil has received limited attention, with some studies indicating these bacteria have little or no ability to persist in soil. Survival and (or) growth of P. luminescens strains in previously sterilized soil, and examination of different soil amendments on their numbers in soil, have not been previously reported. Entomopathogenic P. luminescens (ATCC 29999) and a human clinical isolate (ATCC 43949) were introduced into a soil that had been sterilized by autoclaving, with or without different soil amendments, and bacterial numbers were estimated over time by viable plate count. In the previously sterilized soil receiving no exogenous amendments, numbers fell drastically over a week's time, followed by an increase in numbers by day 30. Treatments involving the addition of calcium carbonate and gelatin or casamino acids to soil usually resulted in higher bacterial numbers. For some sampling dates and soil treatments, there were statistically significant differences between the numbers of the two bacterial strains recovered from soil. The two strains of P. luminescens used in this study were able to survive and grow after being inoculated into previously sterilized soil.Key words: Photorhabdus luminescens, survival, soil.
High production of ligninolytic enzymes from white rot fungi in cereal bran liquid medium
Authors: Pickard M A , Vandertol H , Roman R , Vazquez-Duhalt R
Published: April 2011

White rot fungi from the University of Alberta Mold Herbarium, identified as able to degrade aromatics from a study of PCB metabolism, were examined for production of ligninolytic enzymes. Production of lignin peroxidase, manganese peroxidase, laccase, and veratryl alcohol oxidase were monitored during growth in different media. Good growth but low enzyme production occurred in a glucose - malt extract - yeast extract medium. Media containing 2% cereal bran in 60 mM phosphate buffer supported high levels of laccase production, up to 13 000 U/L in Coriolopsis gallica UAMH 8260 and manganese peroxidase activity up to 1100 U/L in Bjerkandera adusta UAMH 8258. Cereal bran media supported higher laccase production than 2,5-xylidine and higher manganese peroxidase production than a medium containing manganous ion plus veratryl alcohol.Key words: cereal bran, laccase, manganese peroxidase, white rot fungi.
Simultaneous removal of phenol, ortho- and para-cresol by mixed anaerobic consortia
Authors: Hajji K Tawfiki , Lépine F , Bisaillon J -G , Beaudet R
Published: April 2011

Two different anerobic consortia, one removing phenol and ortho- (o-) cresol and the other removing para- (p-) cresol, were cultivated in serum bottles using whey as cosubstrate substitute for proteose peptone. Phenol and p-cresol removal with the phenol-removing consortium were the same with 0.0125% (w/v) whey as with 0.05% proteose peptone. For the other consortium, 8 days were required to decrease the p-cresol concentration from 35 to 2 mg/L with 0.025% whey, while 35 days were required to achieve a similar removal with 0.5% proteose peptone. The two consortia were mixed and cultivated with 0.025% whey. Phenolic compound removal with the mixed consortia was as good as that achieved by each of the two initial consortia against their respective substrates. This removal activity was maintained after several transfers. In a continuous upflow fixed-film reactor, the mixed consortia removed over 98% of 150 mg/L of phenol and 35 mg/L of each o- and p-cresol in the influent at 29°C, with 0.025% whey as cosubstrate. The hydraulic retention time (HRT) was 0.25 day, corresponding to a phenolic compound volumic loading rate of 880 mg/(L of reactor × day). Once the continuous flow reactor achieved constant phenolic compound removal, no intermediates were found in the effluent, while in serum bottles, m-toluic acid, an o-cresol intermediate, accumulated. Measurements of the specific activity for the uptake of different substrates demonstrated the presence of all trophic groups involved in methanogenic fermentation. These activities were, in mg of substrate/(g of volatile suspended solids × day), as follows: 849 ± 25 for the acidogens; 554 ± 15 for the acetogens; 934 ± 37 for the aceticlastic methanogens; and 135 ± 15 for the hydrogenophilic methanogens. Electron micrographs of the mixed consortia showed seven different morphological bacterial types, including Methanotrix-like bacteria.Key words: anaerobic degradation, fixed-film reactor, proteose peptone, whey, phenolic compounds.
Regulation of cellulose-inducible structures of Clostridium cellulovorans
Authors: Blair Benjie G , Anderson Kevin L
Published: April 2011

Scanning electron microscopy was used to detect ultrastructural protuberances on the cellulolytic anaerobe Clostridium cellulovorans. Numerous ultrastructural protuberances were observed on cellulose-grown cells, but few were detected on glucose-, fructose-, cellobiose-, or carboxymethylcellulose (CMC)-grown cells. Formation of these protuberances was detected within 2 h of incubation in cellulose medium, but 4 h incubation was required before numerous structures were observed on the cells. When a soluble carbohydrate or CMC was mixed with cellulose-grown cells, the ultrastructural protuberances could no longer be detected. In fact, no protuberances were observed within 5 min following the addition of glucose, cellobiose, or methylglucose to cellulose-grown cells. The presence of these protuberances corresponded with the binding of the Bandeiraea simplicifolia BSI-B4 isolectin to the cell. Cellulose-grown cells had a greater level of observable lectin binding than cellobiose-grown cells, and lectin binding was not detected on glucose- or fructose-grown cells. In addition, lectin binding ability was lost by cellulose-grown cells following the addition of glucose, fructose, or methylglucose to the cellulose medium. A cellulose-affinity protein fraction expressing cellulase activity was also detected in cell extracts of cellobiose- or cellulose-grown cultures. However, this protein fraction was not detected in extracts of glucose-grown cultures, and was rapidly lost (within 5 min) following the addition of glucose to cellulose-grown cultures. The ability of C. cellulovorans to adhere to cellulose was also affected by the energy substrate, but not in the same manner as the protuberance formation or the cellulase-containing protein fraction. Rather, cellobiose-, cellulose-, and CMC-grown cultures adhered to cellulose, but this adherence was not affected by addition of glucose to the medium. This is the first report that soluble carbohydrates caused the rapid loss of some cellulose-inducible systems of C. cellulovorans.Key words: cellulolytic bacteria, bacterial ultrastructure, polycellulosome, scanning electron microscope, lectin binding, cellulosome.
Sequence analysis of theGluconobacter oxydansRecA protein and construction of arecA-deficient mutant
Authors: Liu Yu-Tien , Chen Chia-Geun , Chao Der-Chiang , Lee Fan , Liao Ching-Len , Sytwu Huey-Kang , Chou Chi-Fu , Ji Dar-Der
Published: April 2011

The deduced amino acid sequence of Gluconobacter oxydans RecA protein shows 75.2, 69.4, and 66.2% homology with those from Aquaspirillum magnetotacticum, Escherichia coli, andPseudomonas aeruginosa, respectively. The amino acid residues essential for function of the recombinase, protease, and ATPase in E. coli recA protein are conserved in G. oxydans. Of 24 amino acid residues believed to be the ATP binding domain of E. coli RecA, 17 are found to be identical in G. oxydans RecA. Interestingly, nucleotide sequence alignment between the SOS box of G. orphans recA gene and those from different microorganisms revealed that all the DNA sequences examined have dyad symmetry that can form a stem-loop structure. A G. oxydans recA-deficient mutant (LCC96) was created by allelic exchange using the cloned recA gene that had been insertionally inactivated by a kanamycin-resistance cassette. Such replacement of the wild-type recA with a kanamycin resistance gene in the chromosome was further verified by Southern hybridization. Phenotypically, the recA-deficient mutant is significantly more sensitive to UV irradiation than the wild-type strain, suggesting that the recA gene of G. oxydans ATCC9324 plays a role in repairing DNA damage caused by UV irradiation. Moreover, the mutant strain is much more plasmid transformable than its parent strain, illustrating that G. oxydans LCC96 could be used as a host to take up the recombinant plasmid for gene manipulation.Key words: Gluconobacter orphans, recA gene, DNA repair, recA mutant, SOS box.
Relationship between H2S-producing strains of wine yeast and different fermentation conditions
Authors: Tamayo C , Ubeda J , Briones A
Published: April 2011

Hydrogen sulphide formation is a problem in winemaking. One of the factors affecting formation of this unwanted metabolite is the yeast strain responsible for the process. In this experiment wines were made on a laboratory scale with different strains of H2S-producing Saccharomyces cerevisiae. The relationship between H2S production and various fermentation conditions was examined (SO2, methionine, (NH4)2SO4, (NH4)3PO4, steel, and steel-lees). The results show that in fermentations in the presence of stainless steel and lees, H2S formation is high but declines when (NH4)3PO4is added to the must.Key words: H2S formation, wine-yeast, steel-lees, wine-making, alcoholic fermentation.
Cross-induction of pyrene and phenanthrene in aMycobacteriumsp. isolated from polycyclic aromatic hydrocarbon contaminated river sediments
Authors: Molina Marirosa , Araujo Rochelle , Hodson Robert E
Published: April 2011

A polycyclic aromatic hydrocarbon (PAH)-degrading culture enriched from contaminated river sediments and a Mycobacterium sp. isolated from the enrichment were tested to investigate the possible synergistic and antagonistic interactions affecting the degradation of pyrene in the presence of low molecular weight PAHs. The Mycobacterium sp. was able to mineralize 63% of the added pyrene when it was present as a sole source of carbon and energy. When the enrichment culture and the isolated bacterium were exposed to phenanthrene, de novo protein synthesis was not required for the rapid mineralization of pyrene, which reached 52% in chloramphenicol-treated cultures and 44% in the absence of the protein inhibitor. In the presence of chloramphenicol, <1% of the added pyrene was mineralized by the mixed culture after exposure to anthracene and naphthalene. These compounds did not inhibit pyrene utilization when present at the same time as pyrene. Concurrent mineralization of pyrene and phenanthrene after exposure to either compound was observed. Cross-acclimation between ring classes of PAHs may be a potentially important interaction influencing the biodegradation of aromatic compounds in contaminated environments.Key words: pyrene, polycyclic aromatic hydrocarbons (PAHs), biodegradation, Mycobacterium sp.
Cloning, sequencing, and expression of cscA invertase from Escherichia coli B-62
Authors: Sahin-Tóth Miklós , Lengyel Zsolt , Tsunekawa Hiroshi
Published: April 2011

We have isolated a 2.5-kb DNA fragment from plasmid pST5R7 encoding a sucrose utilization system from Escherichia coli B-62 which confers a sucrose-fermenting phenotype to transformed E. coli K-12 strains. DNA-sequence determination revealed one full-length open reading frame 98% identical to cscA, the sucrose-hydrolase (invertase) gene of the csc regulon from E. coli EC3132. Functional characterization indicates that high-level expression and limited periplasmic release of invertase is responsible for the sucrose-fermenting capacity of transformed E. coli K-12 strains carrying cscA.Key words: sucrose utilization, sucrose hydrolase, invertase, recombinant protein production.
Subcellular localization of the Streptococcus mutans P1 protein C terminus
Authors: Homonylo-McGavin Mary K , Lee Song F , Bowden George H
Published: April 2011

To determine the subcellular location of the Streptococcus mutans P1 protein C-terminal anchor, cell envelope fractionation experiments were conducted in combination with Western immunoblotting, using monoclonal antibody MAb 6-8C specific for an epitope that maps near the C terminus of P1 protein and also a polyclonal antibody preparation directed against the P1 C-terminal 144 amino acids (P1COOH). P1 protein was detected in cell walls but not the membrane purified from S. mutans cells by the monoclonal antibody. In contrast, P1 protein was not detected in the same cell wall preparation using the anti-P1COOH polyclonal antibody. However, proteins released from the cell walls by treatment with mutanolysin contained antigen that was recognized by the anti-P1COOH antibody, suggesting that the epitopes recognized by the antibody were masked by peptidoglycan in the cell wall preparations. When cell walls were treated with boiling trichloroacetic acid to solubilize cell-wall-associated carbohydrate, P1 antigen could not be detected in either the solubilized carbohydrate, or in the remaining peptidoglycan, regardless of whether polyclonal or monoclonal antibody was used. However, when the peptidoglycan was treated with mutanolysin, P1 antigen could be detected in the mutanolysin solubilized fraction by MAb 6-8C. Collectively, these data suggest that the C-terminal 144 amino acids of the P1 protein are embedded within the cell wall, and associated exclusively with the peptidoglycan. Furthermore, the ability of the anti-P1COOH antibody to recognize P1 antigen only after mutanolysin treatment of cell walls suggests these C-terminal 144 amino acids are tightly intercalated within the peptidoglycan strands.Key words: antigen P1, cell wall proteins, fusion proteins, peptidoglycan, protein localization.
Physiological response ofPectinatus frisingensis,a beer spoilage bacterium,to mild heat treatments
Authors: Tholozan Jean-Luc , Jacquemont Jean
Published: April 2011

Genus Pectinatus is strictly anaerobic bacteria described as a new beer spoilage flora. The physiological response of Pectinatus frisingensis to increasing heat treatments has been studied. Cell death occurred at temperatures higher than 50°C and increased with time. During heat treatment at 50°C, a potassium efflux of more than 50% of the internal potassium was measured at pH 6.2 in starving bacteria, whereas a small transient potassium efflux was measured with a similar 50°C treatment in energized cell suspensions. At beer pH values (pH 4.0), potassium content of P. frisingensis cells was not changed by a moderate heat treatment. Internal pH values of cells were only slightly (0.1 pH unit) decreased upon heat treatments. In contrast, membrane potential value was lowered by a heat treatment at pH 6.2 in deenergized cells, while only a transient decrease of delta was measured with glucose in the medium. A moderate heat treatment at 50°C had no effect on the membrane potential value at pH 4.0, even after 1 h of treatment. In addition, compared with a high level of adenylate energy charge (AEC) measured in energized cell suspensions, an AEC of 0.7 was routinely measured in starving cell suspensions. Moderate heat treatments at pH 4.0 lowered the AEC of cells to 0.6. The physiological response of P. frisingensis to mild heat treatments demonstrated a significant ability of the cell to maintain internal homeostasis at pH conditions encountered in beer.Key words: Pectinatus, thermal death, beer spoilage, homeostasis.
Sphingomonassp. strain Lep1: an aerobic degrader of 4-methylquinoline
Authors: Pfaller Stacy L , Sutton Susan D , Kinkle Brian K
Published: April 2011

Strain Lep1, isolated from a bacterial consortium capable of aerobic degradation of 4-methylquinoline (4-MQ), was chosen for further characterization as it was the only member of the consortium able to grow on 4-MQ in pure culture. Lep1 was identified as a Sphingomonas sp. based on phylogenetic analysis of 16S rDNA. Furthermore, the presence of sphingolipids and 2-hydroxy fatty acids in the membrane, and a 63% G + C ratio supports the placement of Lep1 in this genus. Additional genetic, physiological, and ecological characterization of bacteria such as Lep1 will allow for the potential exploitation of degradative strains for purposes of bioremediation of contaminated soils.Key words: Sphingomonas, degradation, 4-methylquinoline.
The porin OmpC ofSalmonella typhimuriummediates adherence to macrophages
Authors: Negm Robert S , Pistole Thomas G
Published: April 2011

Macrophages recognize, adhere to, and phagocytose Salmonella typhimurium. The major outer membrane protein OmpC is a candidate ligand for macrophage recognition. To confirm this we used transposon mutagenesis to develop an ompC-deficient mutant in a known virulent strain of S. typhimurium; mutant and wild type were compared in macrophage adherence and association assays. Radiolabeled wild type S. typhimurium bound to macrophages at five-fold higher levels than did the ompC mutant. In association assays, macrophages in monolayers bound and internalized three-fold more wild type than mutant, while macrophages in suspension bound and internalized 40-fold more wild type than mutant. The ompC gene of our test strain of S. typhimurium contains several discrete differences compared with the ompC genes of Salmonella typhi and Escherichia coli. The deduced OmpC amino acid sequence of S. typhimurium shares 77 and 98% identity with OmpC amino acid sequences of E. coli and S. typhi, respectively. Evidence from this study supports a role for the OmpC protein in initial recognition by macrophages and distinguishes regions of this protein that potentially participate in host-cell recognition of bacteria by phagocytic cells.Key words: Salmonella, porin, macrophage, outer membrane protein, DNA sequencing.
Glycerol production by yeasts under osmotic and sulfite stress
Authors: Petrovska Blagica , Winkelhausen Eleonora , Kuzmanova Slobodanka
Published: April 2011

The yeasts Saccharomyces cerevisiae,Candida boidinii,Pichia angusta, and Pichia anomala were tested for glycerol production both under osmotic stress and by addition of a sulfite-steering agent. The osmotic pressure was increased by employing glucose concentrations from 50 to 200 g/L and by supplementing with NaCl (40 g/L). Of all the yeasts, S. cerevisiae exhibited the highest level of osmotolerance. The increased osmotic pressure affected glycerol formation the most in C. boidinii. In both Pichia species, glycerol formation was not sufficiently induced when exposed to sugar and salt stress. The addition of 40 g/L Na2SO3 to the medium containing 100 g/L glucose shifted the metabolism of all yeasts towards glycerol formation. Saccharomyces cerevisiae achieved 68.6%, while C. boidinii reached 25.5% of the theoretical glycerol yield, respectively. The highest glycerol yield, 82.3% of the theoretical, was produced by S. cerevisiae under microaerophilic conditions.Key words: glycerol, osmotolerant yeast, sulfite-tolerant yeast, Saccharomyces cerevisiae,Candida boidinii.
The effect of water, ascorbic acid, and cranberry derived supplementation on human urine and uropathogen adhesion to silicone rubber
Authors: Habash Marc B , Van der Mei Henny C , Busscher Henk J , Reid Gregor
Published: April 2011

In this study, urine was collected from groups of volunteers following the consumption of water, ascorbic acid, or cranberry supplements. Only ascorbic acid intake consistently produced acidic urine. Photospectroscopy data indicated that increased water consumption produced urine with lower protein content. Surface tension measurements of the collected urine showed that both water and cranberry supplementation consistently produced urine with surface tensions higher than the control or urine collected following ascorbic acid intake. These urine samples were also employed to study uropathogen adhesion to silicone rubber in a parallel plate flow chamber. Urine obtained after ascorbic acid or cranberry supplementation reduced the initial deposition rates and numbers of adherent Escherichia coli and Enterococcus faecalis, but not Pseudomonas aeruginosa, Staphylococcus epidermidis, or Candida albicans. Conversely, urine obtained from subjects with increased water intake vastly increased the initial deposition rates and numbers of adherent E. coli and E. faecalis (P < 0.05).Key words: adhesion, human urine, natural products, silicone rubber, uropathogens.
Development of a polymerase chain reaction diagnostic test for the detection of the biotrophic pathogen Plasmopara halstedii in sunflower
Authors: Roeckel-Drevet Patricia , Tourvieille Jeanne , Drevet Joël R , Says-Lesage Véronique , Nicolas Paul , de Labrouhe Denis Tourvieille
Published: April 2011

The obligate parasitic fungus-like organism Plasmopara halstedii (Farl.) Berl. et De Toni, is the causal agent of downy mildew disease in sunflower (Helianthus annuus). New races of this economically important parasite are regularly detected throughout the world. In addition, fungicide-resistant isolates have been reported in Europe and North America. These observations of parasite evolution, as well as the risk of propagation of the disease by infected seeds, means that it is necessary to guarantee the absence of Plasmopara halstedii in seed shipments. We report here the development of a rapid assay that can be used to detect infection by Plasmopara halstedii in plant tissues. Based on the nucleotide sequence information obtained from one cloned random amplified polymorphic DNA fragment, specific oligonucleotides were designed and used as primers for in vitro DNA amplification by polymerase chain reaction. An amplification product was detected on agarose gel stained with ethidium bromide when DNA from various Plasmopara halstedii races was tested, whereas no amplified DNA was detected when DNA from other origins was tested, including DNA from the host plant. The sensitivity of the technique was evaluated. The assay successfully reveals the presence of Plasmopara halstedii in infected sunflower plants prior to sporulation.Key words : diagnosis, polymerase chain reaction, SCAR, downy mildew, Helianthus annuus.
Diversity of the internal transcribed spacer of rDNA in morels
Authors: Wipf Daniel , Fribourg Anne , Munch Jean Charles , Botton Bernard , Buscot François
Published: April 2011

The internal transcribed spacer (ITS) of rDNA was analysed by PCR/RFLP in 66 strains belonging to 11 putative species of the Morchellaceae and 3 species of the Discinaceae. For certain taxa, in particular Morchella esculenta, Morchella conica, and Morchella elata, isolates from distant geographical origins were available. Nevertheless, no intraspecific variations of ITS length and restriction was noted. All genera were separated and within Morchella; four putative species could be clearly distinguished in the group of the yellow morels (sectio adnatae). In contrast, no distinction could be obtained between the compared taxa in the group of the black morels (sectio distantes). Sequence analyses of the ITS were performed in one strain of seven putative Morchella. The sequence comparison confirmed the distinctions within the yellow morels and allowed us to separate one putative taxon from the others in black morels. Only the 5,8 gene could be aligned in all sequenced samples. The ITS1 and ITS2 sectors could only be aligned within the black and yellow morels, respectively. The genetic and phylogenetic distances between black and yellow morels, respectively revealed by maximum parsimony and neighbour joining analyses, both confirm the great distance between these species groups and address the question of a separation in the genus Morchella.Key words: Morchellaceae, Discinaceae, ITS.
Epitope mapping of monoclonal antibodies specific for the directly cross-linkedmeso-diaminopimelic acid peptidoglycan found in the anaerobic beer spoilage bacteriumPectinatus cerevisiiphilus
Authors: Ziola Barry , Gares Sheryl L , Lorrain Brandene , Gee Lori , Ingledew W M , Lee Sun Y
Published: April 2011

Nineteen monoclonal antibodies (Mabs) were isolated based on reactivity with disrupted Pectinatus cerevisiiphilus cells. All of the Mabs reacted with cells from which the outer membrane had been stripped by incubation with sodium dodecyl sulphate, suggesting the peptidoglycan (PG) layer was involved in binding. Mab reactivity with purified PG confirmed this. Epitope mapping revealed the Mabs in total recognize four binding sites on the PG. Mabs specific for each of the four sites also bound strongly to disrupted Pectinatus frisingensis, Selenomonas lacticifix, Zymophilus paucivorans, and Zymophilus raffinosivorans cells, but weakly to disrupted Megasphaera cerevisiae cells. No antibody reactivity was seen with disrupted cells of 11 other species of Gram-negative bacteria. These results confirm that a common PG structure is used by several species of anaerobic Gram-negative beer spoilage bacteria. These results also indicate that PG-specific Mabs can be used to rapidly detect a range of anaerobic Gram-negative beer spoilage bacteria, provided the bacterial outer membrane is first removed to allow antibody binding.Key words: beer spoilage, epitope mapping, monoclonal antibodies, Pectinatus, peptidoglycan.
Production of thaxtomin A by Streptomyces scabies strains in plant extract containing media
Authors: Beauséjour Julie , Goyer Claudia , Vachon Joanne , Beaulieu Carole
Published: April 2011

Thaxtomin A production by Streptomyces scabies is induced by plant extracts in culture media. We compared the production of thaxtomin A by three Streptomyces scabies strains (EF-35, CG1, and ATCC 49173) in different culture media in a first attempt to identify the plant molecules required for the biosynthesis of thaxtomin A. Thaxtomin A production varied greatly among media and among S. scabies strains. Strain CG1 presented a higher production than the other strains in all complex media. Oat bran broth is the best thaxtomin A production medium, and suberin is the only plant polymer allowing the production of thaxtomin A in minimal medium. The three S. scabies strains had extracellular esterase activities when grown in the presence of potato suberin or oat bran. We suggest that extracellular esterases allow the release from lipid polymers of plant molecules that may act as inducers for thaxtomin A biosynthesis genes or as precursors in the biosynthetic pathway of thaxtomin A.Key words: common scab, phytotoxin, suberin, esterase.
Impact of an urban effluent on the bacterial community structure in the Arga River (Spain), with special reference to culturable Gram-negative rods
Authors: Goñi-Urriza Marisol , Capdepuy Michèle , Raymond Nathalie , Quentin Claudine , Caumette Pierre
Published: April 2011

The Arga River is an interesting system in which to study the impact of urban effluent pollution because it receives a single effluent in the form of wastewater discharge from the city of Pamplona. To analyze the extent of this discharge, total bacteria, culturable heterotrophic bacteria, and Gram-negative heterotrophic bacteria were enumerated and 409 isolates of the latter were identified. One sampling station was located upstream from the inflow, while five were located up to 30 km downstream. Bacterial counts increased drastically downstream from the wastewater inflow. Total bacterial numbers decreased along the 30 km downstream, the last station attaining similar values to those recorded upstream from the discharge. However, culturable heterotrophic and Gram-negative heterotrophic bacteria levels generally remained significantly higher within the 30 km zone investigated. Among the 409 isolates identified, Aeromonas spp. were the most frequent both upstream and downstream from the discharge. In contrast, although strains belonging to different genera of Enterobacteriaceae were found in all stations, their occurrence was significantly higher just downstream from the polluted discharge. Acinetobacter spp., which were never found upstream, were detected in all stations below the discharge. Our results clearly show that the bacterial community structure changes definitively downstream from the discharge and that Aeromonas were common throughout the sampling zone. Thus they cannot be considered good indicators of pollution in this setting compared to some genera of Enterobacteriaceae or some species ofAcinetobacter, the distribution of which correlated better with the distance from the wastewater discharge.Key words: Aeromonas, Enterobacteriaceae, sewage, freshwater.
Degradation of 2,4-dinitrophenol and selected nitroaromatic compounds bySphingomonassp. UG30
Authors: Zablotowicz R M , Leung K T , Alber T , Cassidy M B , Trevors J T , Lee H , Veldhuis L , Hall J C
Published: April 2011

Sphingomonas strain UG30 mineralizes both p-nitrophenol (PNP) and pentachlorophenol (PCP). Our current studies showed that UG30 oxidatively metabolized certain other p-substituted nitrophenols, i.e., p-nitrocatechol, 2,4-dinitrophenol (2,4-DNP), and 4,6-dinitrocresol with liberation of nitrite. 2,6-DNP, o- or m-nitrophenol, picric acid, or the herbicide dinoseb were not metabolized. Studies using14C-labelled 2,4-DNP indicated that in glucose-glutamate broth cultures of UG30, greater than 90% of 103 µM 2,4-DNP was transformed to other compounds, while 8-19% of the 2,4-DNP was mineralized within 5 days. A significant portion (20-50%) of the 2,4-DNP was metabolized to highly polar metabolite(s) with one major unidentified metabolite accumulating from 5 to 25% of the initial radioactivity. The amounts of 2,4-DNP mineralized and converted to polar metabolites was affected by glutamate concentration in the medium. Nitrophenolic compounds metabolized by UG30 were also suitable substrates for the UG30 PCP-4-monooxygenase (pcpB gene expressed in Escherichia coli) which is likely central to degradation of these compounds. The wide substrate range of UG30 could render this strain useful in bioremediation of some chemically contaminated soils.Key words: bioremediation, dinitrophenol, metabolism, nitroaromatic, pentachlorophenol, Sphingomonas.
Effect of ultrafiltration of yeast extracts on their ability to promote lactic acid bacteria growth
Authors: Gaudreau H , Champagne C P , Conway J , Degré R
Published: April 2011

Five yeast extracts (YE) were fractionated by ultrafiltration (UF) with 1, 3, and 10 kDa molecular weight cutoff membranes, concentrated by freeze-drying, and the resulting powders of yeast extract filtrates (YEF) were evaluated for their growth-promoting properties on nine cultures of lactic acid bacteria (LAB). There was an increase in α-amino nitrogen content of the YEF powders as the pore size of the UF membranes used to filter the YE solutions decreased. The source of YE had a much greater effect than UF on the growth of LAB. This was also the case for the YEF contents in total and α-amino nitrogen. Growth curves of the LAB showed that maximum growth rate (μmax) data were on average 30% higher with bakers' YE than with brewers' YE, while maximum optical density (ODmax) values were on average 16% higher with bakers' YE. This could be related to the higher nitrogen content of the bakers' YE used in this study. Modification by UF of the YE had no significant influence on the growth of 4 of the 9 LAB strains. The three strains of Lactobacillus casei were negatively influenced by UF, as they did not grow as well in the media containing the YEF obtained after filtering with 1 and 3 kDa membranes. On a total solids basis, the 2.5× retentates from the 10 kDa membrane gave, on average, 4% lower μmax and 5% lower ODmax values as compared to cultures where the corresponding YEF was used as medium supplement. This could also be partially related to the different nitrogen contents of the filtrates and retentates. Key words: Lactococcus, Pediococcus, Lactobacillus, amino acids.
Fungi that cause sapstain in Canadian softwoods
Authors: Uzunovic A , Yang D -Q , Gagné P , Breuil C , Bernier L , Byrne A , Gignac M , Kim S H
Published: April 2011

The Canadian forest products industry suffers considerable losses due to discoloration caused by sapstain fungi. Although studied for a number of years, the identity, biology, and ecology of these fungi are still only partly understood. To determine which fungi caused stain problems, a detailed survey was conducted at seven selected sawmills across Canada. In summer 1997, fresh logs and lumber were set aside in the mills 1 month prior to sampling for fungi. We excluded bark-beetle-attacked wood and our logs remained free from their attack during storage. Five commercially important softwood species, Abies balsamea (L.) Mill., Picea mariana (Mill.) B.S.P., Picea glauca (Moench) Voss, Pinus contorta Dougl., and Pinus banksiana Lamb., were included in the studies. A total of 1863 isolates were isolated from stained and adjacent areas on test wood, and were identified based on their morphological and physiological characteristics and mating compatibility to 13 different species representing five genera. The most commonly encountered genus, Ophiostoma (97%), was represented by nine species. A more diverse range of fungi was found in logs than in lumber; some species were more frequently isolated from one type of substrate and rarely (or not at all) from the other. No fungal species occurred exclusively in a particular region or wood substrate.Key words: bluestain, lumber, logs, Ophiostoma, Ceratocystis.
Characterization of aNeocallimastixpatriciarumxylanase gene and its product
Authors: Liu Jin-Hao , Selinger Brent L , Tsai Cheng-Fang , Cheng Kuo-Jaon
Published: April 2011

A xylanase gene (xynC) isolated from the anaerobic ruminal fungus Neocallimastix patriciarum was characterized. The gene consists of an N-terminal catalytic domain that exhibited homology to family 11 of glycosyl hydrolases, a C-terminal cellulose binding domain (CBD) and a putative dockerin domain in between. Each domain was linked by a short linker domain rich in proline and alanine. Deletion analysis demonstrated that the CBD was essential for optimal xylanase activity of the enzyme, while the putative dockerin domain may not be required for enzyme function.Key words: xylanase, cellulose binding domain, Neocallimastix patriciarum.
Sulfur cycling in mixed cultures of sulfide-oxidizing and sulfate- or sulfur-reducing oil field bacteria
Authors: Telang Anita J , Jenneman Gary E , Voordouw Gerrit
Published: April 2011

Thiomicrospira strain CVO and Arcobacter strain FWKO B are two nitrate-reducing, sulfide-oxidizing bacteria (NR-SOB) isolated from the Coleville field in western Canada. Analysis by reverse sample genome probing (RSGP) indicates that both can be enriched from Coleville produced water samples by addition of nitrate. Neither could be enriched from waters produced from oil fields with a high resident temperature (40-60°C). In co-cultures of the two microorganisms, CVO dominated at lower and FWKO B at higher sulfide concentrations. Sulfide concentrations of up to 15 mM (480 ppm) could be controlled (i.e., reduced to lower values) by the co-culture. Mixed cultures of a sulfate-reducing bacterium (Desulfovibrio strain Lac6) and CVO or FWKO B produced considerably less sulfide than cultures of Lac6 alone, indicating that these NR-SOB can efficiently oxidize sulfide generated by sulfate reduction provided nitrate is present. The same observations were made for mixed cultures of a sulfur-oxidizing bacterium (Desulfuromonas acetoxidans) and CVO. The results indicate that successful nitrate-mediated conversion of sulfide to oxidized forms (sulfur, sulfate) in oil field production waters by NR-SOB requires the absence of high concentrations of electron donors (e.g., lactate, acetate) that can reverse the nitrate-mediated oxidation reaction.