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Pseudomonas Fluorescens (pseudomona + fluorescen)

Distribution by Scientific Domains

Life Sciences	50%
Chemistry	35%
Earth and Environmental Science	8%
2 Other Domains	7%

Selected Abstracts

Critical aspects of analysis of Micrococcus luteus, Neisseria cinerea, and Pseudomonas fluorescens by means of capillary electrophoresis

ELECTROPHORESIS, Issue 18-19 2004
Verena Hoerr
Abstract Within the frame of our study we investigated Microccocus luteus, Neisseria cinerea, and Pseudomonas fluorescens by means of capillary zone electrophoresis (CZE). They form chains and clusters on a different scale, which can be reflected in the electropherograms. A low buffer concentration of Tris-borate and Na2 EDTA containing a polymeric matrix of 0.0125% poly(ethylene) oxide (PEO) was used. Key factors were the standardization and optimization of CE conditions, buffer solution, and pretreatment of bacterial samples, which are not transferable to different bacterial strains, in general. The different compositions of the cell wall of on the one hand Gram-positive (M. luteus) and Gram-negative (N. cinerea) cocci and on the other hand Gram-negative, rod-shaped bacteria (P.fluorescens), are probably responsible for the different pretreatment conditions. [source]

Pseudomonas fluorescens orchestrates a fine metabolic-balancing act to counter aluminium toxicity

ENVIRONMENTAL MICROBIOLOGY, Issue 6 2010
Joseph Lemire
Summary Aluminium (Al), an environmental toxin, is known to disrupt cellular functions by perturbing iron (Fe) homeostasis. However, Fe is essential for such metabolic processes as the tricarboxylic acid (TCA) cycle and oxidative phosphorylation, the two pivotal networks that mediate ATP production during aerobiosis. To counter the Fe conundrum induced by Al toxicity, Pseudomonas fluorescens utilizes isocitrate lyase and isocitrate dehydrogenase-NADP dependent to metabolize citrate when confronted with an ineffective aconitase provoked by Al stress. By invoking fumarase C, a hydratase devoid of Fe, this microbe is able to generate essential metabolites. To compensate for the severely diminished enzymes like Complex I, Complex II and Complex IV, the upregulation of a H2O-generating NADH oxidase enables the metabolism of citrate, the sole carbon source via a modified TCA cycle. The overexpression of succinyl-CoA synthetase affords an effective route to ATP production by substrate-level phosphorylation in the absence of O2. This fine metabolic balance enables P. fluorescens to survive the dearth of bioavailable Fe triggered by an Al environment, a feature that may have potential applications in bioremediation technologies. [source]

Biofilm formation and cellulose expression among diverse environmental Pseudomonas isolates

ENVIRONMENTAL MICROBIOLOGY, Issue 11 2006
Susanne Ude
Summary The ability to form biofilms is seen as an increasingly important colonization strategy among both pathogenic and environmental bacteria. A survey of 185 plant-associated, phytopathogenic, soil and river Pseudomonas isolates resulted in 76% producing biofilms at the air,liquid (A,L) interface after selection in static microcosms. Considerable variation in biofilm phenotype was observed, including waxy aggregations, viscous and floccular masses, and physically cohesive biofilms with continuously varying strengths over 1500-fold. Calcofluor epifluorescent microscopy identified cellulose as the matrix component in biofilms produced by Pseudomonas asplenii, Pseudomonas corrugata, Pseudomonas fluorescens, Pseudomonas marginalis, Pseudomonas putida, Pseudomonas savastanoi and Pseudomonas syringae isolates. Cellulose expression and biofilm formation could be induced by the constitutively active WspR19 mutant of the cyclic-di-GMP-associated, GGDEF domain-containing response regulator involved in the P. fluorescens SBW25 wrinkly spreader phenotype and cellular aggregation in Pseudomonas aeruginosa PA01. WspR19 could also induce P. putida KT2440, which otherwise did not produce a biofilm or express cellulose, as well as Escherichia coli K12 and Salmonella typhimurium LT2, both of which express cellulose yet lack WspR homologues. Statistical analysis of biofilm parameters suggest that biofilm development is a more complex process than that simply described by the production of attachment and matrix components and bacterial growth. This complexity was also seen in multivariate analysis as a species-ecological habitat effect, underscoring the fact that in vitro biofilms are abstractions of those surface and volume colonization processes used by bacteria in their natural environments. [source]

Cadmium-regulated gene fusions in Pseudomonas fluorescens

ENVIRONMENTAL MICROBIOLOGY, Issue 4 2000
Silvia Rossbach
To study the mechanisms soil bacteria use to cope with elevated concentrations of heavy metals in the environment, a mutagenesis with the lacZ -based reporter gene transposon Tn5 -B20 was performed. Random gene fusions in the genome of the common soil bacterium Pseudomonas fluorescens strain ATCC 13525 were used to create a bank of 5000 P. fluorescens mutants. This mutant bank was screened for differential gene expression in the presence of the toxic metal cadmium. Fourteen mutants were identified that responded with increased or reduced gene expression to the presence of cadmium. The mutants were characterized with respect to their metal-dependent gene expression and their metal tolerance. Half the identified mutants reacted with differential gene expression specifically to the metal cadmium, whereas some of the other mutants also responded to elevated concentrations of copper and zinc ions. One of the mutants, strain C8, also showed increased gene expression in the presence of the solvent ethanol, but otherwise no overlap between cadmium-induced gene expression and general stress response was detected. Molecular analysis of the corresponding genetic loci was performed using arbitrary polymerase chain reaction (PCR), DNA sequencing and comparison of the deduced protein products with sequences deposited in genetic databases. Some of the genetic loci targeted by the transposon did not show any similarities to any known genes; thus, they may represent ,novel' loci. The hypothesis that genes that are differentially expressed in the presence of heavy metals play a role in metal tolerance was verified for one of the mutants. This mutant, strain C11, was hypersensitive to cadmium and zinc ions. In mutant C11, the transposon had inserted into a genetic region displaying similarity to genes encoding the sensor/regulator protein pairs of two-component systems that regulate gene expression in metal-resistant bacteria, including czcRS of Ralstonia eutropha, czrRS of Pseudomonas aeruginosa and copRS of Pseudomonas syringae. Although the P. fluorescens strain used in this study had not been isolated from a metal-rich environment, it nevertheless contained at least one genetic region enabling it to cope with elevated concentrations of heavy metals. [source]

Chemotactic response of plant-growth-promoting bacteria towards roots of vesicular-arbuscular mycorrhizal tomato plants

FEMS MICROBIOLOGY ECOLOGY, Issue 3 2003
Sushma Gupta Sood
Abstract The chemotactic responses of the plant-growth-promoting rhizobacteria Azotobacter chroococcum and Pseudomonas fluorescens to roots of vesicular-arbuscular mycorrhizal (Glomus fasciculatum) tomato plants were determined. A significantly (P=0.05) greater number of bacterial cells of wild strains were attracted towards vesicular-arbuscular mycorrhizal tomato roots compared to non-vesicular-arbuscular mycorrhizal tomato roots. Substances exuded by roots served as chemoattractants for these bacteria. P. fluorescens was strongly attracted towards citric and malic acids, which were predominant constituents in root exudates of tomato plants. A. chroococcum showed a stronger response towards sugars than amino acids, but the response was weakest towards organic acids. The effects of temperature, pH, and soil water matric potential on bacterial chemotaxis towards roots were also investigated. In general, significantly (P=0.05) greater chemotactic responses of bacteria were observed at higher water matric potentials (0, ,1, and ,5 kPa), slightly acidic to neutral pH (6, 6.5 and 7), and at 20,30°C (depending on the bacterium) than in other environmental conditions. It is suggested that chemotaxis of P. fluorescens and A. chroococcum towards roots and their exudates is one of the several steps in the interaction process between bacteria and vesicular-arbuscular mycorrhizal roots. [source]

Porins of Pseudomonas fluorescens MFO as fibronectin-binding proteins

FEMS MICROBIOLOGY LETTERS, Issue 1 2002
J. Rebière-Huët
Abstract Bacterial adherence is a complex phenomenon involving specific interactions between receptors, including matricial fibronectin, and bacterial ligands. We show here that fibronectin and outer membrane proteins of Pseudomonas fluorescens were able to inhibit adherence of P. fluorescens to fibronectin-coated wells. We identified at least six fibronectin-binding proteins with molecular masses of 70, 55, 44, 37, 32 and 28 kDa. The presence of native (32 kDa) and heat-modified forms (37 kDa) of OprF was revealed by immuno-analysis and the 44-kDa band was composed of three proteins, their N-terminal sequences showing homologies with Pseudomonas aeruginosa porins (OprD, OprE1 and OprE3). [source]

Elevated zinc induces siderophore biosynthesis genes and a zntA -like gene in Pseudomonas fluorescens

FEMS MICROBIOLOGY LETTERS, Issue 1 2000
Silvia Rossbach
Abstract Zinc-regulated genes were analyzed in Pseudomonas fluorescens employing mutagenesis with a reporter gene transposon. Six mutants responded with increased gene expression to elevated concentrations of zinc. Genetic and biochemical analysis revealed that in four of the six mutants the transposon had inserted into genes essential for the biosynthesis of the siderophore pyoverdine. The growth of one of the mutants was severely impaired in the presence of elevated concentrations of cadmium and zinc ions. In this mutant, the transposon had inserted in a gene with high similarity to P-type ATPases involved in zinc and cadmium ion transport. Four mutants reacted with reduced gene expression to elevated concentrations of zinc. One of these mutants was sensitive to zinc, cadmium and copper ions. The genetic region targeted in this mutant did not show similarity to any known gene. [source]

Mutualistic symbiosis between Bursaphelenchus xylophilus and bacteria of the genus Pseudomonas

FOREST PATHOLOGY, Issue 5 2005
B. G. Zhao
Summary Interactions between the pine wood nematode (PWN), Bursaphelenchus xylophilus, and bacteria of the genus Pseudomonas were examined by cultivating axenic PWN and bacterial strains using callus of Pinus thunbergii. Ten (Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas cepacia and Pseudomonas spp.) of the 29 bacterial strains tested, significantly increased the reproduction of PWN. The rest of the bacteria (19 strains of 10 species) inhibited the reproduction of PWN completely. The growth of 18 of the 29 bacterial strains tested, including the 10 strains promoting PWN reproduction, was significantly increased by the presence of PWN. It indicated a mutualistic symbiotic relationship between PWN and the 10 bacterial strains in the genus Pseudomonas. The bacterial mutualistic symbionts are organisms, which may have co-evolved with PWN rather than being accidentally associated. The finding provides further evidence for our hypothesis that pine wilt disease is complex, induced by both PWN and associated phytotoxin-producing bacteria. Résumé Les interactions entre le nématode des pins Bursaphelenchus xylophilus (PWN) et des bactéries du genre Pseudomonas ont étéétudiées en cultivant de manière axénique PWN et des souches bactériennes sur des cals de Pinusthunbergii. Dix souches bactériennes (Pseudomonas fluorescens, P. putida, P. cepacia et Pseudomonas spp.) sur les 29 testées ont significativement augmenté la reproduction de PNW. Le reste des bactéries (19 souches de 10 espèces) ont complètement inhibé la reproduction de PNW. La croissance de 18 souches bactériennes sur 29, incluant les 10 favorisant la reproduction de PNW, a été significativement augmentée en présence de PNW. Ceci indique une relation symbiotique mutualiste entre PNW et 10 souches bactériennes du genre Pseudomonas. Les symbiontes bactériens mutualistes pourraient être des organismes ayant coévolué avec PNW plutôt qu'associés de façon fortuite. Ces observations renforcent l'hypothèse selon laquelle le flétrissement des pins est une maladie complexe induite par PWN en association avec des bactéries productrices de phytotoxines. Zusammenfassung Die Interaktionen zwischen dem Kiefernsplintholznematoden (PWN, Bursaphelenchus xylophilus) und Bakterien der Gattung Pseudomonas wurden in axenischen Kulturen des Nematoden mit verschiedenen Bakterienstämmen und Kallus von Pinus thunbergii untersucht. Zehn Bakterienstämme (Pseudomonas fluoreszens, Pseudomonas putida, Pseudomonas cepacia und Pseudomonas spp.) von 29 getesteten Isolaten erhöhten die Reproduktion des PWN signifikant. Die übrigen Isolate (19 Stämme von 10 Arten) hemmten die Vermehrung des Nematoden vollständig. Das Wachstum von 18 der 29 getesteten Bakterienstämme (inkl. der 10 Stämme, welche die Vermehrung des Nematoden förderten), wurde durch die Präsenz des Nematoden signifikant erhöht. Dieser Befund deutet auf eine mutualistische Beziehung zwischen dem PWN und 10 Pseudomonas -Isolaten hin. Die mutualistischen bakteriellen Symbionten dürften sich wahrscheinlich in Coevolution mit dem Nematoden entwickelt haben. Dieser Befund unterstützt die Hypothese, dass die Kiefernwelke eine Komplexkrankheit darstellt, die sowohl durch B. xylophilus als auch die damit assoziierten phytotoxinbildenden Bakterien ausgelöst wird. [source]

Mobilization of metals from uranium mine waste: the role of pyoverdines produced by Pseudomonas fluorescens

GEOBIOLOGY, Issue 4 2010
F. EDBERG
Microorganisms produce chelating agents, such as siderophores and other ligands, which allow them to mobilize and scavenge essential elements from the environment when bioavailability is low. To better understand the effects of biologically mediated leaching of metals from mine waste, Pseudomonas fluorescens was cultivated in the presence of processed ore from the former uranium mine in Ranstad, southern Sweden. Light conditions, the concentration of the mineral source and oxygen availability were varied. The presence of ore in the culture flasks enhanced bacterial growth and raised the pH of the culture medium. Increasing the amount of ore or enhancing aeration of the medium further encouraged cell growth and pH rise. Bacteria mobilized Fe, Ni and Co from the ore. Fe-siderophore complexes were detected and estimated to be present at approximately 9 ,m. In the presence of bacteria and light, dissolved Fe and U concentrations were higher compared to dark conditions. Increasing the amount of ore resulted in higher dissolved Ni concentrations but lower dissolved Fe, most likely due to precipitate formation. Data from this study support siderophore production by bacteria that allowed mobilization of essential nutrients from the processed ore. However, the availability of potentially toxic metals like Ni and U may also be enhanced. Microbial-promoted mobilization could contribute to leaching of toxic metals in current and historic mining areas. This process should be considered during design and implementation of remediation projects where trace metals are of environmental concern. [source]

Degradation of naphthenic acids by sediment micro-organisms

JOURNAL OF APPLIED MICROBIOLOGY, Issue 5 2006
L.F. Del Rio
Abstract Aims:, Naphthenic acids (NAs) are naturally occurring, linear and cyclic carboxylic surfactants associated with the acidic fraction of petroleum. NAs account for most of the acute aquatic toxicity of oil sands process-affected water (OSPW). The toxicity of OSPW can be reduced by microbial degradation. The aim of this research was to determine the extent of NA degradation by sediment microbial communities exposed to varying amounts of OSPW. Methods and Results:, Eleven wetlands, both natural and process-affected, and one tailings settling pond in Northern Alberta were studied. The natural wetlands and process-affected sites fell into two distinct groups based on their water chemistry. The extent of degradation of a 14C-labelled monocyclic NA surrogate [14C-cyclohexane carboxylic acid (CCA)] was relatively uniform in all sediments (approximately 30%) after 14 days. In contrast, degradation of a bicyclic NA surrogate [14C-decahydronaphthoic acid (DHNA)]was significantly lower in non process-affected sediments. Enrichment cultures, obtained from an active tailings settling pond, using commercially available NAs as the sole carbon source, resulted in the isolation of a co-culture containing Pseudomonas putida and Pseudomonas fluorescens. Quantitative GC,MS analysis showed that the co-culture removed >95% of the commercial NAs, and partially degraded the process NAs from OSPW with a resulting NA profile similar to that from ,aged wetlands'. Conclusions:, Exposure to NAs induced and/or selected micro-organisms capable of more effectively degrading bicyclic NAs. Native Pseudomonas spp. extensively degraded fresh, commercial NA. The recalcitrant NAs resembled those found in process-affected wetlands. Significance and Impact of the Study:, These results suggest that it may be possible to manipulate the existing environmental conditions to select for a microbial community exhibiting higher rates of NA degradation. This will have significant impact on the design of artificial wetlands for water treatment. [source]

The role of host organism, transcriptional switches and reporter mechanisms in the performance of Hg-induced biosensors

JOURNAL OF APPLIED MICROBIOLOGY, Issue 6 2004
M. Harkins
Abstract Aims:, The purpose of this study was to comprehensively compare the response of nine biosensors capable of being induced by Hg. Induction by Hg was based upon the insertion of merR, merB, zntA and zntR promoter genes. LuxCDABE or lucFF reporter genes expressed luminescence, and host organisms were Escherichia coli, Vibrio anguillarum and Pseudomonas fluorescens. The role of transcriptional switches, reporter mechanism and host organism was to be investigated. Methods and Results:, All biosensors were subjected to the same assay conditions. Sensors had their own individual growth characteristics and response to the doses of Hg tested. Maximum bioluminescence response was induced by concentrations of Hg between 2·5 nm and 5 ,m. E. coli pRB28 was found to detect levels of Hg as low as 1·6 nm and yet was capable of operating in a concentration range of up to 12·5 ,m. Conclusions:, The response of the sensors demonstrated their suitability for analysis under environmentally relevant concentrations. The sensitivity of the sensors, the optimum range and the expediency of the assay could not be related to a single sensor trait. It may be concluded that biosensor performance is dependent on more than one of the single factors studied. Significance and Impact of the Study:, The results show that comparative testing of sensors is an important step in evaluating the relevance and performance of biosensors prior to routine environmental application. [source]

Performance characteristics of cholesterol oxidase for kinetic determination of total cholesterol

JOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 6 2005
Pornpen Srisawasdi
Abstract The enzymatic method for cholesterol determination can use either an endpoint or a kinetic method. Not much is known concerning the properties (Km and Vmax) of the commercial enzyme for the kinetic method. We measured the Km and Vmax of Brevibacterium, Streptomyces, Pseudomonas fluorescens, and Cellulomonas cholesterol oxidase. Brevibacterium gave the highest Km value (230.3×10,4,M), followed by Streptomyces (2.17×10,4,M), Cellulomonas (0.84×10,4,M), and Pseudomonas (0.61×10,4,M). The Km values and the linearity obtained from Streptomyces (2.6,mmol/L), Pseudomonas (2.1,mmol/L), or Cellulomonas (2.1,mmol/L) were too low. Dichlorophenol isomers, acting as inhibitors, increased the enzyme's Km. The addition of 3,4-dichlorophenol raised the Km of Streptomyces from 2.17×10,4 to 24.89×10,4,M. The linearity was increased from 2.6 to 13.0,mmol/L. The high Km of Brevibacterium resulted in an insensitive reaction and low cholesterol linearity (7.8,mmol/L). An increase in the sample-to-reagent ratio from 1:100 to 1:10 enhanced the reaction rate and the linearity from 7.8 to 20.7,mmol/L. We suggest that Brevibacterium and Streptomyces cholesterol oxidase (with the addition of 3,4 dichlorophenol) are good sources for serum cholesterol determination by the kinetic method. J. Clin. Lab. Anal. 19:247,252, 2005. © 2005 Wiley-Liss, Inc. [source]

MHC-linked susceptibility to a bacterial infection, but no MHC-linked cryptic female choice in whitefish

JOURNAL OF EVOLUTIONARY BIOLOGY, Issue 1 2004
C. Wedekind
Abstract Non-random gamete fusion is one of several potential cryptic female choice mechanisms that have been postulated and that may enhance the survival probability of the offspring. Previous studies have found that gamete fusion in mice is influenced by genes of the major histocompatibility complex (MHC) region. Here we test (i) whether there is MHC-dependent gamete fusion in whitefish (Coregonus sp.) and (ii) whether there is a link between the MHC and embryo susceptibility to an infection by the bacterium Pseudomonas fluorescens. We experimentally bred whitefish and reared sibships in several batches that either experienced or did not experience strong selection by P. fluorescens. We then determined the MHC class II B1 genotype of 1016 surviving larvae of several full sibships. We found no evidence for MHC-linked gamete fusion. However, in one of seven sibships we found a strong connection between the MHC class II genotype and embryo susceptibility to P. fluorescens. This connection was still significant after correcting for multiple testing. Hence, the MHC class II genotype can considerably influence embryo survival in whitefish, but gamete fusion seems to be random with respect to the MHC. [source]

MICROBIOLOGICAL AND PATHOGENIC CONTAMINANTS OF SEAFOOD IN GREECE

JOURNAL OF FOOD QUALITY, Issue 1 2007
C. PAPADOPOULOU
ABSTRACT A total of 360 samples, including 105 marine fish, 25 prawns, 50 squid, 50 octopus, 30 mussels and 100 freshwater fish were examined microbiologically for the presence of microorganisms potentially pathogenic. All samples were examined following standard microbiological procedures. The isolated microorganisms were: Aeromonas hydrophilia (38,93%), Klebsiella ozonae (1,40%), Escherichia coli (14,87%), Yersinia enterocolitica (0,40%), Hafnia alvei (0,36.6%), Enterobacter agglomerans (0,42%), Citrobacter freundii (0,46%), Proteus vulgaris (15,80%), Proteus mirabilis (7,82%), Morganella morganii (0,30%), Pseudomonas fluorescens (0,34%), Pseudomonas putida (0,6%), Plesiomonas shigelloides (0,4%), Listeria innocua (1,3.3%), Vibrio parahaemolyticus (0,2%), Clostridium perfringens (0,1%), Staphylococcus aureus (0,80%) and Candida quillermondi (0,1%), Candida albicans (0,1%), Penicillium oxalicum (0,1%) and Penicillium italicum (0,12%). [source]

ANTIBACTERIAL ACTIVITY AND BINDING ABILITY OF BOVINE LACTOFERRIN AGAINST PSEUDOMONAS SPP.

JOURNAL OF FOOD SAFETY, Issue 1 2008
WOAN-SUB KIM
ABSTRACT The antibacterial activity of bovine lactoferrin was tested against Pseudomonas fluorescens and Pseudomonas syringae. The activity was studied by monitoring the growth of a Pseudomonas spp. in the presence or absence of bovine apo-lactoferrin, bovine holo-lactoferrin or native-lactoferrin in liquid media at different concentrations. Lactoferrin-binding proteins in the membrane fractions of Pseudomonas spp. were detected using far-Western blot analysis. The addition of bovine lactoferrin to the medium inhibited the growth of all tested strains. Furthermore, the growth of P. fluorescens and P. syringae was strongly inhibited by bovine apo-lactoferrin. The estimated molecular weights of lactoferrin-binding proteins in P. fluorescens were 70, 49, 47 and 25 kDa, and 70, 48 and 28 kDa in P. syringae. PRACTICAL APPLICATIONS Pseudomonas fluorescens is an important psychrotrophic bacterium responsible for undesirable flavors in milk and dairy products. Thus, flavor and texture defects, such as bitterness and running paste, were also reported. In addition, Pseudomonas syringae causes various diseases on many different susceptible plant species, generally producing chlorotic and necrotic lesions on leaves and fruits. The resultant bacterial spoilage causes considerable economic losses for the food and dairy industries. At present, antiseptics and agricultural chemicals are used for defense of foods and vegetables from these bacteria, but such substances are known to deleteriously affect the human body. The results of this study demonstrate that bovine lactoferrin significantly inhibits the growth of P. fluorescens and P. syringae. The results indicate that the incorporation of bovine lactoferrin is expected to protect dairy products, food and fruits from pathogenic bacteria. [source]

Use of Bean Sprout Enterobacteriaceae Isolates as Biological Control Agents of Pseudomonas fluorescens

JOURNAL OF FOOD SCIENCE, Issue 1 2004
K. ENOMOTOArticle first published online: 28 JUN 200
ABSTRACT: Bean sprouts were cultivated under in vitro conditions as a model system to study the mechanism of bacteria-mediated spoilage in bean sprouts. Pseudomonas fluorescens, or Erwinia spp., were inoculated onto sprouts at several stages during cultivation. Five strains of Enterobacteriaceae isolated from the native microflora of sprouts prevented Pseudomonas -mediated spoilage by co-inoculating these cultures on seeds that were soaking in water. The population of P. fluorescens in co-inoculated liquid medium culture with a strain (B1) decreased slightly. The results indicated that the Enterobacteriaceae isolates tested played an important role in preventing Pseudomonas -mediated spoilage by growing competitively with P. fluorescens. [source]

Microbial and Sensory Assessment of Milk with an Electronic Nose

JOURNAL OF FOOD SCIENCE, Issue 2 2002
F. Korel
ABSTRACT: An electronic nose (e-nose) was used to assess milk odor inoculated with Pseudomonas fluorescens or Bacillus coagulans, and odors were correlated with microbial loads and sensory scores. Sterile whole, reduced-fat, and fat-free milk were inoculated, stored at 1.7, 7.2, and 12.8 °C, and evaluated at d 0, 3, 5, 7, and 10 by e-nose and sensory panel. Aerobic plate counts were performed. E-nose readings, microbial counts, and sensory data were analyzed using discriminant function analysis. The e-nose discriminated differences in odor due to microbial load and sensory data. This may lead to a rapid method for determining sensory evaluation and microbial loads of milk. [source]

Development of Biogenic Amines in Yellowfin Tuna (Thunnus albacares): Effect of Storage and Correlation with Decarboxylase-Positive Bacterial Flora

JOURNAL OF FOOD SCIENCE, Issue 1 2002
W.-X. Du
ABSTRACT: The effects of storage at 0,4,10, and 22°C for 0,1,3,5, and 9 d on the quality of yellowfin tuna fillets as determined by microbiological assessment, development of some biogenic amines, and sensory analysis were studied. Tuna fillets stored at 22 °C for 3 d, 10 °C for 5 d, and 4 °C for 9 d were rated unacceptable for consumption. Those stored at 22 °C for 3 d had total aerobic bacterial count of > 8 log10 CFU/g, a histamine-producing bacterial population of 7 log10 CFU/g, and 832 ppm of histamine, 35.8 ppm of putrescine, and 147 ppm of cadaverine. A comparison of the capillary electrophoresis, AOAC fluorometric method, and gas chromatography showed a very good correlation (r2 > 0.99) among these 3 methods for histamine quantitation in tuna samples. Morganella morganii, Enterobacter agglomerans, Enterobacter intermedium, Pseudomonas fluorescens, Proteins vulgaris, and Serratia liquefaciens were the decarboxylase-positive bacterial species isolated by using the Niven's medium and identified during storage, which were responsible for histamine production in test tuna fillets. [source]

Inactivation Kinetics of Foodborne Spoilage and Pathogenic Bacteria by Ozone

JOURNAL OF FOOD SCIENCE, Issue 3 2000
J.-G. Kim
ABSTRACT: Ozone was tested against Pseudomonas fluorescens, Escherichia coli O157:H7, Leuconostoc mesenteroides, and Listeria monocytogenes. When kinetic data from a batch reactor were fitted to a dose-response model, a 2-phased linear relationship was observed. A continuous ozone reactor was developed to ensure a uniform exposure of bacterial cells to ozone and a constant concentration of ozone during the treatment. Survivors plots in the continuous system were linear initially, followed by a concave downward pattern. Exposure of bacteria to ozone at 2.5 ppm for 40 s caused 5 to 6 log decrease in count. Resistance of tested bacteria to ozone followed this descending order: E. coli O157:H7, P. fluorescens, L. mesenteroides, and L. monocytogenes. [source]

Antibacterial activities and total phenolic contents of grape pomace extracts

JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 14 2004
Gülcan Özkan
Abstract The aim of this study was to determine the total phenolic contents and antibacterial effects of grape pomace extracts (cultivars Emir and Kalecik karasi) against 14 bacteria, and the effects of the extracts on the growth and survival of two of the bacteria during storage. The total phenolic contents of grape pomace of Emir and Kalecik karasi cultivars extracted with acetone/water/acetic acid (90:9.5:0.5) were 68.77 and 96.25 mg GAE g,1, respectively. The agar well diffusion method was used to test the antibacterial activity of the extracts at 1, 2.5, 5, 10 and 20% (w/v) concentrations in methanol on spoilage and pathogenic bacteria including Aeromonas hydrophila, Bacillus cereus, Enterobacter aerogenes, Enterococcus faecalis, Escherichia coli, Escherichia coli O157:H7. Mycobacterium smegmatis, Proteus vulgaris, Pseudomonas aeruginosa, Pseudomonas fluorescens, Salmonella enteritidis, Salmonella typhimurium, Staphylococcus aureus and Yersinia enterocolitica. All the bacteria tested were inhibited by extract concentrations of 2.5, 5, 10 and 20%, except for Y enterocolitica which was not inhibited by the 2.5% concentration. However, pomace extracts at 1% concentration had no antibacterial activity against some of the bacteria. According to the agar well diffusion method, E coli O157:H7 was the most sensitive of the bacteria. Generally, using the serial dilution method, while the extracts at 0.5% concentration had bacteriostatic activities on E coli O157:H7 and S aureus, the extracts appeared to have bactericidal effects at 1 and 2.5% concentrations. In accordance with this method, S aureus was more sensitive than E coli O157:H7 to the extracts. Copyright © 2004 Society of Chemical Industry [source]

Synthesis of Polycaprolactone Using Free/Supported Enzymatic and Non-Enzymatic Catalysts

MACROMOLECULAR RAPID COMMUNICATIONS, Issue 24 2004
María Laura Foresti
Abstract Summary: Polymerization of caprolactone using lipases from Candida antarctica B, Rhizomucor meihei, Candida rugosa, and Pseudomonas fluorescens is highly effective, with 97% conversion into polycaprolactone. Poly(propylene)-supported Candida rugosa lipase achieves higher conversion values (85,92%) than free lipase (75%). Acidic and basic non-conventional catalysis with butanol yields 50,85% conversion. Simple UV/visible techniques gave the same results for measuring conversion than other studies. Applications are opened for the non-conventional catalysts. Mechanism of the polymerization of caprolactone polymerization using a basic catalyst. [source]

Role of 2,4-Diacetylphloroglucinol-Producing Fluorescent Pseudomonas spp. in the Defense of Plant Roots

PLANT BIOLOGY, Issue 1 2007
D. M. Weller
Abstract: Plants have evolved strategies of stimulating and supporting specific groups of antagonistic microorganisms in the rhizosphere as a defense against diseases caused by soilborne plant pathogens owing to a lack of genetic resistance to some of the most common and widespread soilborne pathogens. Some of the best examples of natural microbial defense of plant roots occur in disease suppressive soils. Soil suppressiveness against many different diseases has been described. Take-all is an important root disease of wheat, and soils become suppressive to take-all when wheat or barley is grown continuously in a field following a disease outbreak; this phenomenon is known as take-all decline (TAD). In Washington State, USA and The Netherlands, TAD results from the enrichment during monoculture of populations of 2,4-diacetylphloroglucinol (2,4-DAPG)-producing Pseudomonas fluorescens to a density of 105 CFU/g of root, the threshold required to suppress the take-all pathogen, Gaeumannomyces graminis var. tritici. 2,4-DAPG-producing P. fluorescens also are enriched by monoculture of other crops such as pea and flax, and evidence is accumulating that 2,4-DAPG producers contribute to the defense of plant roots in many different agroecosystems. At this time, 22 distinct genotypes of 2,4-DAPG producers (designated A - T, PfY and PfZ) have been defined by whole-cell repetitive sequence-based (rep)-PCR analysis, restriction fragment length polymorphism (RFLP) analysis of phlD, and phylogenetic analysis of phlD, but the number of genotypes is expected to increase. The genotype of an isolate is predictive of its rhizosphere competence on wheat and pea. Multiple genotypes often occur in a single soil and the crop species grown modulates the outcome of the competition among these genotypes in the rhizosphere. 2,4-DAPG producers are highly effective biocontrol agents against a variety of plant diseases and ideally suited for serving as vectors for expressing other biocontrol traits in the rhizosphere. [source]

In vitro antibacterial effect of berberine hydrochloride and enrofloxacin to fish pathogenic bacteria

AQUACULTURE RESEARCH, Issue 7 2010
Defeng Zhang
Abstract Berberine the main antibacterial substance in the rhizome of Coptis chinensis Franch, is used as an antimicrobial agent for a long time. This study was carried out to investigate the potential of berberine alone or in combination with enrofloxacin against six common fish pathogens for use in fish disease management in aquaculture. The minimum inhibitory concentrations (MICs) of berberine hydrochloride against Aeromonas hydrophila, Pseudomonas fluorescens, Vibrio vulnificus, Edwardsiella ictaluri, Escherichia coli and Streptococcus agalactiae were determined and to be >500, >500, >500, 300, 400 and 100 ,g mL,1 respectively. The minimal bactericidal concentrations (MBCs) of berberine hydrochloride to E. coli, E. ictaluri and S. agalactiae were 300,500 ,g mL,1. In combination with enrofloxacin, the MICs and MBCs of berberine hydrochloride significantly decreased against E. coli and E. ictaluri but not against Streptococcus dysgalactiae. These results demonstrated that the berberine hydrochloride enhanced the bactericidal effect of enrofloxacin and vice versa. The synergistic bactericidal effect of berberine hydrochloride and enrofloxacin suggest its potential use in fish disease management in aquaculture. This is the first study on the inhibitory effect of berberine hydrochloride against fish pathogenic bacteria. [source]

Effect of periplasmic nitrate reductase on diauxic lag of Paracoccus pantotrophus

BIOTECHNOLOGY PROGRESS, Issue 4 2009
Kiranmai Durvasula
Abstract Paracoccus pantotrophus expresses two nitrate reductases,membrane bound nitrate reductase (Nar) and periplasmic nitrate reductase (Nap). In growth experiments with two denitrifying species (Paracoccus pantotrophus and Alcaligenes eutrophus) that have both Nap and Nar and two species (Pseudomonas denitrificans and Pseudomonas fluorescens) with Nar only, it was found that diauxic lag is shorter for bacteria that express Nap. In P. pantotrophus, napEDABC encodes the periplasmic nitrate reductase. To analyze the effect of Nap on diauxic lag, the nap operon was deleted from P. pantotrophus. The growth experiments with nap, mutant resulted in increased diauxic lag when switched from aerobic to anoxic respiration, suggesting Nap is responsible for shorter lags and helps in adaptation to anoxic metabolism after transition from aerobic conditions. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009 [source]

Use of Glycol Ethers for Selective Release of Periplasmic Proteins from Gram-Negative Bacteria

BIOTECHNOLOGY PROGRESS, Issue 5 2007
Jeffrey R. Allen
Genetic modification of Gram-negative bacteria to express a desired protein within the cellapos;s periplasmic space, located between the inner cytoplasmic membrane and the outer cell wall, can offer an attractive strategy for commercial production of therapeutic proteins and industrial enzymes. In certain applications, the product expression rate is high, and the ability to isolate the product from the cell mass is greatly enhanced because of the productapos;s compartmentalized location within the cell. Protein release methods that increase the permeability of the outer cell wall for primary recovery, but avoid rupturing the inner cell membrane, reduce contamination of the recovered product with other host cell components and simplify final purification. This article reports representative data for a new release method employing glycol ether solvents. The example involves the use of 2-butoxyethanol (commonly called ethylene glycol n -butyl ether or EB) for selective release of a proprietary biopharmaceutical protein produced in the periplasmic space of Pseudomonas fluorescens. In this example, glycol ether treatment yielded ,65% primary recovery with ,80% purity on a protein-only basis. Compared with other methods including heat treatment, osmotic shock, and the use of surfactants, the glycol ether treatment yielded significantly reduced concentrations of other host cell proteins, lipopolysaccharide endotoxin, and DNA in the recovered protein solution. The use of glycol ethers also allowed exploitation of temperature-change-induced phase splitting behavior to concentrate the desired product. Heating the aqueous EB extract solution to 55 °C formed two liquid phases: a glycol ether-rich phase and an aqueous product phase containing the great majority of the product protein. This phase-splitting step yielded an approximate 10-fold reduction in the volume of the initial product solution and a corresponding increase in the productapos;s concentration. [source]

Crystallization, diffraction data collection and preliminary crystallographic analysis of DING protein from Pseudomonas fluorescens

ACTA CRYSTALLOGRAPHICA SECTION F (ELECTRONIC), Issue 7 2007
Sebastien Moniot
PfluDING is a phosphate-binding protein expressed in Pseudomonas fluorescens. This protein is clearly distinct from the bacterial ABC transporter soluble phosphate-binding protein PstS and is more homologous to eukaryotic DING proteins. Interestingly, bacterial DING proteins have only been detected in certain Pseudomonas species. Although DING proteins seem to be ubiquitous in eukaryotes, they are systematically absent from eukaryotic genomic databases and thus are still quite mysterious and poorly characterized. PfluDING displays mitogenic activity towards human cells and binds various ligands such as inorganic phosphate, pyrophosphate, nucleotide triphosphates and cotinine. Here, the crystallization of PfluDING is reported in a monoclinic space group (P21), with typical unit-cell parameters a = 36.7, b = 123.7, c = 40.8,Å, , = 90, , = 116.7, , = 90°. Preliminary crystallographic analysis reveals good diffraction quality for these crystals and a 1.43,Å resolution data set has been collected. [source]

Natural Diversity to Guide Focused Directed Evolution

CHEMBIOCHEM, Issue 13 2010
Helge Jochens Dr.
Abstract Simultaneous multiple site-saturation mutagenesis was performed at four active-site positions of an esterase from Pseudomonas fluorescens to improve its ability to convert 3-phenylbutyric acid esters (3-PBA) in an enantioselective manner. Based on an appropriate codon choice derived from a structural alignment of 1751 sequences of ,/,-hydrolase fold enzymes, only those amino acids were considered for library creation that appeared frequently in structurally equivalent positions. Thus, the number of mutants to be screened could be substantially reduced while the number of functionally intact variants was increased. Whereas the wild-type esterase showed only marginal activity and poor enantioselectivity (Etrue=3.2) towards 3-PBA-ethyl ester, a significant number of hits with improved rates (up to 240-fold) and enantioselectivities (up to Etrue=80) were identified in these "smart" libraries. [source]

Factors Mediating Activity, Selectivity, and Substrate Specificity for the Thiamin Diphosphate-Dependent Enzymes Benzaldehyde Lyase and Benzoylformate Decarboxylase

CHEMBIOCHEM, Issue 12 2006
Michael Knoll
Abstract Benzaldehyde lyase from Pseudomonas fluorescens and benzoylformate decarboxylase from Pseudomonas putida are homologous thiamin diphosphate-dependent enzymes that catalyze carboligase and carbolyase reactions. Both enzymes catalyze the formation of chiral 2-hydroxy ketones from aldehydes. However, the reverse reaction has only been observed with benzaldehyde lyase. Whereas benzaldehyde lyase is strictly R specific, the stereoselectivity of benzoylformate decarboxylase from P. putida is dependent on the structure and orientation of the substrate aldehydes. In this study, the binding sites of both enzymes were investigated by using molecular modelling studies to explain the experimentally observed differences in the activity, stereo- and enantioselectivity and substrate specificity of both enzymes. We designed a detailed illustration that describes the shape of the binding site of both enzymes and sufficiently explains the experimental effects observed with the wild-type enzymes and different variants. These findings demonstrate that steric reasons are predominantly responsible for the differences observed in the (R)-benzoin cleavage and in the formation of chiral 2-hydroxy ketones. [source]

Pseudomonas fluorescens' view of the periodic table

ENVIRONMENTAL MICROBIOLOGY, Issue 1 2008
Matthew L. Workentine
Summary Growth in a biofilm modulates microbial metal susceptibility, sometimes increasing the ability of microorganisms to withstand toxic metal species by several orders of magnitude. In this study, a high-throughput metal toxicity screen was initiated with the aim of correlating biological toxicity data in planktonic and biofilm cells to the physiochemical properties of metal ions. To this end, Pseudomonas fluorescens ATCC 13525 was grown in the Calgary Biofilm Device (CBD) and biofilms and planktonic cells of this microorganism were exposed to gradient arrays of different metal ions. These arrays included 44 different metals with representative compounds that spanned every group of the periodic table (except for the halogens and noble gases). The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and minimum biofilm eradication concentration (MBEC) values were obtained after exposing the biofilms to metal ions for 4 h. Using these values, metal ion toxicity was correlated to the following ion-specific physicochemical parameters: standard reduction-oxidation potential, electronegativity, the solubility product of the corresponding metal,sulfide complex, the Pearson softness index, electron density and the covalent index. When the ions were grouped according to outer shell electron structure, we found that heavy metal ions gave the strongest correlations to these parameters and were more toxic on average than the other classes of the ions. Correlations were different for biofilms than for planktonic cells, indicating that chemical mechanisms of metal ion toxicity differ between the two modes of growth. We suggest that biofilms can specifically counter the toxic effects of certain physicochemical parameters, which may contribute to the increased ability of biofilms to withstand metal toxicity. [source]