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Sample records for pseudomonas fluorescens alginate

  1. 40 CFR 180.1114 - Pseudomonas fluorescens A506, Pseudomonas fluorescens 1629RS, and Pseudomonas syringae 742RS...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Pseudomonas fluorescens A506, Pseudomonas fluorescens 1629RS, and Pseudomonas syringae 742RS; exemptions from the requirement of a tolerance... Tolerances § 180.1114 Pseudomonas fluorescens A506, Pseudomonas fluorescens 1629RS, and Pseudomonas syringae...

  2. Growth of Pseudomonas fluorescens on Cassava Starch ...

    African Journals Online (AJOL)

    Michael Horsfall

    ABSTRACT: The potential of local strains of microorganism (Pseudomonas fluorescens) in polyhydroxbutyrate production ... The demand for the use of biopolymers ... This work therefore investigates the production of polyhydroxybutyrate from.

  3. Water flow induced transport of Pseudomonas fluorescens cells through soil columns as affected by inoculant treatment

    NARCIS (Netherlands)

    Hekman, W.E.; Heijnen, C.E.; Trevors, J.T.; Elsas, van J.D.

    1994-01-01

    Water flow induced transport of Pseudomonas fluorescens cells through soil columns was measured as affected by the inoculant treatment. Bacterial cells were introduced into the topsoil of columns, either encapsulated in alginate beads of different types or mixed with bentonite clay in concentrations

  4. Pseudomonas fluorescens' view of the periodic table.

    Science.gov (United States)

    Workentine, Matthew L; Harrison, Joe J; Stenroos, Pernilla U; Ceri, Howard; Turner, Raymond J

    2008-01-01

    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.

  5. Detection of Pseudomonas fluorescens from broth, water and ...

    African Journals Online (AJOL)

    Loop mediated isothermal amplification is rapid, highly sensitive and specifically developed method for detection of bacterial infections. AprX gene for alkaline metalloprotease of Pseudomonas fluorescens was used to design four primers and loop mediated isothermal amplification (LAMP) conditions were standardized for ...

  6. ADHESION OF PSEUDOMONAS-FLUORESCENS TO METALLIC SURFACES

    NARCIS (Netherlands)

    VIEIRA, MJ; OLIVEIRA, R; MELO, L; PINHEIRO, M; VANDERMEI, HC

    1992-01-01

    Deposition of Pseudomonas fluorescens on aluminium, brass and copper plates was studied in a flow system. The number of bacteria deposited on aluminium was greater than on the other two types of metals. The results are discussed in terms of the mechanisms (transport and/or adhesion) that may control

  7. Genome Sequence of the Biocontrol Strain Pseudomonas fluorescens F113

    Science.gov (United States)

    Redondo-Nieto, Miguel; Barret, Matthieu; Morrisey, John P.; Germaine, Kieran; Martínez-Granero, Francisco; Barahona, Emma; Navazo, Ana; Sánchez-Contreras, María; Moynihan, Jennifer A.; Giddens, Stephen R.; Coppoolse, Eric R.; Muriel, Candela; Stiekema, Willem J.; Rainey, Paul B.; Dowling, David; O'Gara, Fergal; Martín, Marta

    2012-01-01

    Pseudomonas fluorescens F113 is a plant growth-promoting rhizobacterium (PGPR) that has biocontrol activity against fungal plant pathogens and is a model for rhizosphere colonization. Here, we present its complete genome sequence, which shows that besides a core genome very similar to those of other strains sequenced within this species, F113 possesses a wide array of genes encoding specialized functions for thriving in the rhizosphere and interacting with eukaryotic organisms. PMID:22328765

  8. Lethality and Developmental Delay of Drosophila melanogaster Following Ingestion of Selected Pseudomonas fluorescens Strains

    Science.gov (United States)

    Pseudomonas fluorescens secretes antimicrobial compounds that promote plant health and provide protection from pathogens. We used a non-invasive feeding assay to study the toxicity of P. fluorescens strains Pf0-1, SBW25, and Pf-5 to Drosophila melanogaster. The three strains of P. fluorescens varie...

  9. Production of yellow-green fluorescent pigment by Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    Gildo Almeida da Silva

    2006-05-01

    Full Text Available A medium was prepared from brewery waste yeast with and without mineral salts to study growth and yellow-green fluorescent pigment production (YGFP by Pseudomonas fluorescens. The King's medium used for detection of siderophore production were expressively weaker inductors of YGFP formation when compared to FYE medium. Although FYE and CYE could be used for growth of P. fluorescens, only FYE was an attractive medium for detection of YGFP strain producers.Diversos microrganismos secretam substâncias com alta afinidade por ferro. Estas moléculas, sideróforos, transportam ferro para o interior das células. Como a produção destas moléculas depende da composição do meio, foi avaliada a influência do extrato de levedura (FYE, proveniente de resíduo de cervejaria, com e sem adição de sais minerais, sobre o crescimento de Pseudomonas fluorescens e sobre a formação de pigmento fluorescente verde-amarelado (YGFP. Observou-se que (i FYE com sacarose (G7 e o extrato de levedura comercial (CYE possuem um pico bem definido próximo a 260 nm; (ii FYE, mas não CYE, promove alta formação de YGFP. Os meios de King's, usados para detectar a formação de sideróforo, se comportaram como indutores expressivamente mais fracos de YGFP que o meio FYE. Embora FYE e CYE possam ser usados para o crescimento de P. fluorescens, apenas FYE pode ser usado como meio para a detecção de linhagens formadoras de YGFP.

  10. 40 CFR 180.1200 - Pseudomonas fluorescens strain PRA-25; temporary exemption from the requirement of a tolerance.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Pseudomonas fluorescens strain PRA-25... RESIDUES IN FOOD Exemptions From Tolerances § 180.1200 Pseudomonas fluorescens strain PRA-25; temporary... established for residues of the microbial pesticide, pseudomonas fluorescens strain PRA-25 when used on peas...

  11. Factors Affecting Zebra Mussel Kill by the Bacterium Pseudomonas fluorescens

    Energy Technology Data Exchange (ETDEWEB)

    Daniel P. Molloy

    2004-02-24

    The specific purpose of this research project was to identify factors that affect zebra mussel kill by the bacterium Pseudomonas fluorescens. Test results obtained during this three-year project identified the following key variables as affecting mussel kill: treatment concentration, treatment duration, mussel siphoning activity, dissolved oxygen concentration, water temperature, and naturally suspended particle load. Using this latter information, the project culminated in a series of pipe tests which achieved high mussel kill inside power plants under once-through conditions using service water in artificial pipes.

  12. Combined inoculation of Pseudomonas fluorescens and Trichoderma harzianum for enhancing plant growth of vanilla (Vanilla planifolia).

    Science.gov (United States)

    Sandheep, A R; Asok, A K; Jisha, M S

    2013-06-15

    This study was conducted to evaluate the plant growth promoting efficiency of combined inoculation of rhizobacteria on Vanilla plants. Based on the in vitro performance of indigenous Trichoderma spp. and Pseudomonas spp., four effective antagonists were selected and screened under greenhouse experiment for their growth enhancement potential. The maximum percentage of growth enhancement were observed in the combination of Trichoderma harzianum with Pseudomonas fluorescens treatment followed by Pseudomonas fluorescens, Trichoderma harzianum, Pseudomonas putida and Trichoderma virens, respectively in decreasing order. Combined inoculation of Trichoderma harzianum and Pseudomonas fluorescens registered the maximum length of vine (82.88 cm), highest number of leaves (26.67/plant), recorded the highest fresh weight of shoots (61.54 g plant(-1)), fresh weight of roots (4.46 g plant(-1)) and dry weight of shoot (4.56 g plant(-1)) where as the highest dry weight of roots (2.0806 g plant(-1)) were achieved with treatments of Pseudomonas fluorescens. Among the inoculated strains, combined inoculation of Trichoderma harzianum and Pseudomonas fluorescens recorded the maximum nitrogen uptake (61.28 mg plant(-1)) followed by the combined inoculation of Trichoderma harzianum (std) and Pseudomonas fluorescens (std) (55.03 mg plant(-1)) and the highest phosphorus uptake (38.80 mg plant(-1)) was recorded in dual inoculation of Trichoderma harzianum and Pseudomonas fluorescens.

  13. Foam separation of Pseudomonas fluorescens and Bacillus subtilis var. niger.

    Science.gov (United States)

    Grieves, R B; Wang, S L

    1967-01-01

    An experimental investigation established the effect of the presence of inorganic salts on the foam separation of Pseudomonas fluorescens and of Bacillus subtilis var. niger (B. globigii) from aqueous suspension by use of a cationic surfactant. For P. fluorescens, 5.0 mueq/ml of NaCl, KCl, Na(2)SO(4), K(2)SO(4), CaCl(2), CaSO(4), MgCl(2), or MgSO(4) produced increases in the cell concentration in the residual suspension (not carried into the foam) from 2.9 x 10(5) up to 1.6 x 10(6) to 2.8 x 10(7) cells per milliliter (initial suspensions contain from 3.3 x 10(7) to 4.8 x 10(7) cells per milliliter). The exceptional influence of magnesium was overcome by bringing the cells into contact first with the surfactant and then the salt. For B. subtilis, the presence of 5.0 mueq/ml of any of the eight salts increased the residual cell concentration by one order of magnitude from 1.2 x 10(4) to about 4.0 x 10(5) cells per milliliter. This occurred regardless of the sequence of contact as long as the surfactant contact period was sufficient. The presence of salts increased collapsed foam volumes with P. fluorescens and decreased collapsed foam volumes with B. subtilis.

  14. Effect of Pseudomonas fluorescens on Buried Steel Pipeline Corrosion.

    Science.gov (United States)

    Spark, Amy J; Law, David W; Ward, Liam P; Cole, Ivan S; Best, Adam S

    2017-08-01

    Buried steel infrastructure can be a source of iron ions for bacterial species, leading to microbiologically influenced corrosion (MIC). Localized corrosion of pipelines due to MIC is one of the key failure mechanisms of buried steel pipelines. In order to better understand the mechanisms of localized corrosion in soil, semisolid agar has been developed as an analogue for soil. Here, Pseudomonas fluorescens has been introduced to the system to understand how bacteria interact with steel. Through electrochemical testing including open circuit potentials, potentiodynamic scans, anodic potential holds, and electrochemical impedance spectroscopy it has been shown that P. fluorescens increases the rate of corrosion. Time for oxide and biofilms to develop was shown to not impact on the rate of corrosion but did alter the consistency of biofilm present and the viability of P. fluorescens following electrochemical testing. The proposed mechanism for increased corrosion rates of carbon steel involves the interactions of pyoverdine with the steel, preventing the formation of a cohesive passive layer, after initial cell attachment, followed by the formation of a metal concentration gradient on the steel surface.

  15. Genomic and Genetic Diversity within the Pseudomonas fluorescens Complex.

    Directory of Open Access Journals (Sweden)

    Daniel Garrido-Sanz

    Full Text Available The Pseudomonas fluorescens complex includes Pseudomonas strains that have been taxonomically assigned to more than fifty different species, many of which have been described as plant growth-promoting rhizobacteria (PGPR with potential applications in biocontrol and biofertilization. So far the phylogeny of this complex has been analyzed according to phenotypic traits, 16S rDNA, MLSA and inferred by whole-genome analysis. However, since most of the type strains have not been fully sequenced and new species are frequently described, correlation between taxonomy and phylogenomic analysis is missing. In recent years, the genomes of a large number of strains have been sequenced, showing important genomic heterogeneity and providing information suitable for genomic studies that are important to understand the genomic and genetic diversity shown by strains of this complex. Based on MLSA and several whole-genome sequence-based analyses of 93 sequenced strains, we have divided the P. fluorescens complex into eight phylogenomic groups that agree with previous works based on type strains. Digital DDH (dDDH identified 69 species and 75 subspecies within the 93 genomes. The eight groups corresponded to clustering with a threshold of 31.8% dDDH, in full agreement with our MLSA. The Average Nucleotide Identity (ANI approach showed inconsistencies regarding the assignment to species and to the eight groups. The small core genome of 1,334 CDSs and the large pan-genome of 30,848 CDSs, show the large diversity and genetic heterogeneity of the P. fluorescens complex. However, a low number of strains were enough to explain most of the CDSs diversity at core and strain-specific genomic fractions. Finally, the identification and analysis of group-specific genome and the screening for distinctive characters revealed a phylogenomic distribution of traits among the groups that provided insights into biocontrol and bioremediation applications as well as their role as

  16. ANOMALOUS BLUE COLOURING OF MOZZARELLA CHEESE INTENTIONALLY CONTAMINATED WITH PIGMENT PRODUCING STRAINS OF PSEUDOMONAS FLUORESCENS

    Directory of Open Access Journals (Sweden)

    P. Sechi

    2011-04-01

    Full Text Available In summer 2010 a large outbreak of anomalous blue coloration of mozzarella cheese was recorded in Italy and some northern European countries. Official laboratory analysis and health authorities linked the outbreak to the contamination of processing water with strains of Pseudomonas fluorescens, although several expert raised the question of how to unequivocally link the blue coloring to the presence of the micro-organism. In an attempt to set-up a method to determine whether a given Pseudomonas spp. strain is responsible of the defect, an in vitro system for the evaluation of blue colouring of mozzarella cheese intentionally contaminated with strains of Pseudomonas fluorescens. was developed The system is aimed to ascertain whether P. fluorescens strains, isolated from mozzarella cheese with anomalous blue coloration, are able to reproduce the blue coloration under controlled experimental condition. 96 trials of experimental inoculation of mozzarella cheese in different preservation liquids, were conducted using various suspension of Pseudomonas spp. (P. fluorescens ATCC 13525, P. fluorescens CFBP 3150, one P. fluorescens field strain isolated from blue-colored mozzarella cheese and P. aeruginosa ATCC 10145 as positive control at different concentrations and incubated at different temperatures. Growth curve of all Pseudomonas spp. strains tested demonstrated that after three days of incubation the concentration was generally higher than 106 CFU/g of mozzarella cheese incubated in Tryptic Soy Broth (TSB, and higher than 105 CFU/g of mozzarella cheese incubated in preservation liquid. All mozzarella cheeses inoculated with the field strain of Pseudomonas fluorescens showed the characteristic anomalous blue coloration, which is often associated with Pseudomonas fluorescens contamination of water used during mozzarella cheesemaking. With the proposed system, which enabled a considerable amount of samples to be analysed under controlled experimental

  17. High pressure inactivation of Pseudomonas in black truffle - comparison with Pseudomonas fluorescens in tryptone soya broth

    Science.gov (United States)

    Ballestra, Patricia; Verret, Catherine; Cruz, Christian; Largeteau, Alain; Demazeau, Gerard; El Moueffak, Abdelhamid

    2010-03-01

    Pseudomonas is one of the most common genera in black Perigord truffle. Its inactivation by high pressure (100-500 MPa/10 min) applied on truffles at sub-zero or low temperatures was studied and compared with those of Pseudomonas fluorescens in tryptone soya broth. Pressurization of truffles at 300 MPa/4 °C reduced the bacterial count of Pseudomonas by 5.3 log cycles. Higher pressures of 400 or 500 MPa, at 4 °C or 20 °C, allowed us to slightly increase the level of destruction to the value of ca. 6.5 log cycles but did not permit us to completely inactivate Pseudomonas. The results showed a residual charge of about 10 CFU/g. Pressure-shift freezing of truffles, which consists in applying a pressure of 200 MPa/-18 °C for 10 min and then quickly releasing this pressure to induce freezing, reduced the population of Pseudomonas by 3.3 log cycles. The level of inactivation was higher than those obtained with conventional freezing. Endogenous Pseudomonas in truffle was shown to be more resistant to high pressure treatments than P. fluorescens used for inoculation of broths.

  18. Boolean models of biosurfactants production in Pseudomonas fluorescens.

    Directory of Open Access Journals (Sweden)

    Adrien Richard

    Full Text Available Cyclolipopeptides (CLPs are biosurfactants produced by numerous Pseudomonas fluorescens strains. CLP production is known to be regulated at least by the GacA/GacS two-component pathway, but the full regulatory network is yet largely unknown. In the clinical strain MFN1032, CLP production is abolished by a mutation in the phospholipase C gene (plcC and not restored by plcC complementation. Their production is also subject to phenotypic variation. We used a modelling approach with Boolean networks, which takes into account all these observations concerning CLP production without any assumption on the topology of the considered network. Intensive computation yielded numerous models that satisfy these properties. All models minimizing the number of components point to a bistability in CLP production, which requires the presence of a yet unknown key self-inducible regulator. Furthermore, all suggest that a set of yet unexplained phenotypic variants might also be due to this epigenetic switch. The simplest of these Boolean networks was used to propose a biological regulatory network for CLP production. This modelling approach has allowed a possible regulation to be unravelled and an unusual behaviour of CLP production in P. fluorescens to be explained.

  19. Effects of cysteine on growth, protease production, and catalase activity of Pseudomonas fluorescens.

    OpenAIRE

    Himelbloom, B H; Hassan, H M

    1986-01-01

    Cysteine inhibits growth of and protease production by Pseudomonas fluorescens NC3. Catalase activity in P. fluorescens NC3 was increased by cysteine. The addition of exogenous hydrogen peroxide did not increase catalase activity, thus suggesting a role for the endogenous generation of hydrogen peroxide via the autoxidation of cysteine.

  20. Three Strains of Pseudomonas fluorescens Exhibit Differential Toxicity Against Drosophila melanogaster

    Science.gov (United States)

    Three strains of Pseudomonas fluorescens were tested for toxicity to Drosophila melanogaster in an insect feeding assay. Insect eggs were placed on the surface of a non-nutritive agar plate supplemented with a food source that was non-inoculated or inoculated with P. fluorescens Pf0-1, SBW25, or Pf-...

  1. Adaptive synonymous mutations in an experimentally evolved Pseudomonas fluorescens population

    DEFF Research Database (Denmark)

    Bailey, Susan; Hinz, Aaron; Kassen, Rees

    2014-01-01

    Conventional wisdom holds that synonymous mutations, nucleotide changes that do not alter the encoded amino acid, have no detectable effect on phenotype or fitness. However, a growing body of evidence from both comparative and experimental studies suggests otherwise. Synonymous mutations have been...... shown to impact gene expression, protein folding and fitness, however, direct evidence that they can be positively selected, and so contribute to adaptation, is lacking. Here we report the recovery of two beneficial synonymous single base pair changes that arose spontaneously and independently...... in an experimentally evolved population of Pseudomonas fluorescens. We show experimentally that these mutations increase fitness by an amount comparable to non-synonymous mutations and that the fitness increases stem from increased gene expression. These results provide unequivocal evidence that synonymous mutations...

  2. Bioreporter pseudomonas fluorescens HK44 immobilized in a silica matrix

    Directory of Open Access Journals (Sweden)

    Trogl J.

    2003-01-01

    Full Text Available The bioluminescent bioreporter Pseudomonas fluorescens HK44, the whole cell bacterial biosensor that responds to naphthalene and its metabolites via the production of visible light, was immobilized into a silica matrix by the sol-gel technique. The bioluminescence intensities were measured in the maximum of the bioluminescence band at X = 500 nm. The immobilized cells (>105 cells per g silica matrix produced light after induction by salicylate (cone. > 10 g/l, naphthalene and aminobenzoic acid. The bioluminescence intensities induced by 2,3-dihydroxynaphthalene 3-hydroxybenzoic acid and 4-hydroxybenzoic acid were comparable to a negative control. The cells in the silica layers on glass slides produced light in response to the presence of an inductor at least 8 months after immobilization, and >50 induction cycles. The results showed that these test slides could be used as assays for the multiple determination of water pollution.

  3. Promotion of plant growth by Pseudomonas fluorescens strain SS101 via novel volatile organic compounds

    NARCIS (Netherlands)

    Park, Yong-Soon; Dutta, Swarnalee; Ann, Mina; Raaijmakers, Jos M.; Park, Kyungseok

    2015-01-01

    Abstract Volatile organic compounds (VOCs) from plant growth-promoting rhizobacteria (PGPR) play key roles in modulating plant growth and induced systemic resistance (ISR) to pathogens. Despite their significance, the physiological functions of the specific VOCs produced by Pseudomonas fluorescens

  4. Pseudomonas putida and Pseudomonas fluorescens Species Group Recovery from Human Homes Varies Seasonally and by Environment.

    Directory of Open Access Journals (Sweden)

    Susanna K Remold

    Full Text Available By shedding light on variation in time as well as in space, long-term biogeographic studies can help us define organisms' distribution patterns and understand their underlying drivers. Here we examine distributions of Pseudomonas in and around 15 human homes, focusing on the P. putida and P. fluorescens species groups. We describe recovery from 10,941 samples collected during up to 8 visits per home, occurring on average 2.6 times per year. We collected a mean of 141 samples per visit, from sites in most rooms of the house, from the surrounding yards, and from human and pet occupants. We recovered Pseudomonas in 9.7% of samples, with the majority of isolates being from the P. putida and P. fluorescens species groups (approximately 62% and 23% of Pseudomonas samples recovered respectively. Although representatives of both groups were recovered from every season, every house, and every type of environment sampled, recovery was highly variable across houses and samplings. Whereas recovery of P. putida group was higher in summer and fall than in winter and spring, P. fluorescens group isolates were most often recovered in spring. P. putida group recovery from soils was substantially higher than its recovery from all other environment types, while higher P. fluorescens group recovery from soils than from other sites was much less pronounced. Both species groups were recovered from skin and upper respiratory tract samples from healthy humans and pets, although this occurred infrequently. This study indicates that even species that are able to survive under a broad range of conditions can be rare and variable in their distributions in space and in time. For such groups, determining patterns and causes of stochastic and seasonal variability may be more important for understanding the processes driving their biogeography than the identity of the types of environments in which they can be found.

  5. Expression analysis of the gacS mutant of Pseudomonas fluorescens SBW25

    NARCIS (Netherlands)

    Cheng, Xu; Bruijn, de Irene; Voort, van der M.; Raaijmakers, Jos

    2013-01-01

    Pseudomonas species are ubiquitous in plant-associated environments and produce an array of volatiles, enzymes and antimicrobials. The biosynthesis of many metabolites is regulated by the GacS/GacA two-component regulatory system. Transcriptome analysis of Pseudomonas fluorescens SBW25 revealed that

  6. Enhanced extracellular chitinase production in Pseudomonas fluorescens: biotechnological implications

    Directory of Open Access Journals (Sweden)

    Azhar Alhasawi

    2017-06-01

    Full Text Available Chitin is an important renewable biomass of immense commercial interest. The processing of this biopolymer into value-added products in an environmentally-friendly manner necessitates its conversion into N-acetyl glucosamine (NAG, a reaction mediated by the enzyme chitinase. Here we report on the ability of the soil microbe Pseudomonas fluorescens to secrete copious amounts of chitinase in the spent fluid when cultured in mineral medium with chitin as the sole source of carbon and nitrogen. Although chitinase was detected in various cellular fractions, the enzyme was predominantly localized in the extracellular component that was also rich in NAG and glucosamine. Maximal amounts of chitinase with a specific activity of 80 µmol NAG produced mg–1 protein min–1 was obtained at pH 8 after 6 days of growth in medium with 0.5 g of chitin. In-gel activity assays and Western blot studies revealed three isoenzymes. The enzyme had an optimal activity at pH 10 and a temperature range of 22–38 ℃. It was stable for up to 3 months. Although it showed optimal specificity toward chitin, the enzyme did readily degrade shrimp shells. When these shells (0.1 g were treated with the extracellular chitinase preparation, NAG [3 mmoles (0.003 g-mol] was generated in 6 h. The extracellular nature of the enzyme coupled with its physico-chemical properties make this chitinase an excellent candidate for biotechnological applications.

  7. IDENTIFIKASI SURFAKTIN PADA PSEUDOMONAS FLUORESCENS ST1 PENGENDALI EFEKTIF PENYAKIT PUSTUL KEDELAI

    Directory of Open Access Journals (Sweden)

    Suskandini Ratih Dirmawati .

    2011-11-01

    Full Text Available Identification of surfactin in Pseudomonas fluorescens ST1 which effectively suppres soybean bacterial pustule.   Identification of surfactin in Pseudomonas fluorescens ST1 filtrate was conducted in Plant Pest and Disease Laboratory, Bogor Agriculture University.  The 48 hours cultured suspension of  P. fluorescens ST1 with 108 CFU/ml density was centrifuged to obtain the supernatant.  The supernatant was analyzed for its surfactin content by High Performance Liquid Chromatography with Colum ODS-5 and eluen acetonitril and acetat acid.  The result showed  that  surfactin was producted by P. fluorescens ST1 and this bioactive substance could suppres the bacterial pustule on soybean.

  8. Expression, Purification, Crystallization and Preliminary X-ray Analysis of Pseudomonas fluorescens AlgK

    Energy Technology Data Exchange (ETDEWEB)

    Keiski,C.; Yip, P.; Robinson, H.; Burrows, L.; Howell, P.

    2007-01-01

    AlgK is an outer-membrane lipoprotein involved in the biosynthesis of alginate in Pseudomonads and Azotobacter vinelandii. A recombinant form of Pseudomonas fluorescens AlgK with a C-terminal polyhistidine affinity tag has been expressed and purified from the periplasm of Escherichia coli cells and diffraction-quality crystals of AlgK have been grown using the hanging-drop vapour-diffusion method. The crystals grow as flat plates with unit-cell parameters a = 79.09, b = 107.85, c = 119.15 {angstrom}, = 96.97{sup o}. The crystals exhibit the symmetry of space group P2{sub 1} and diffract to a minimum d-spacing of 2.5 {angstrom} at Station X29 of the National Synchrotron Light Source, Brookhaven National Laboratory. On the basis of the Matthews coefficient (V{sub M} = 2.53 {angstrom}{sup 3} Da{sup -1}), four protein molecules are estimated to be present in the asymmetric unit.

  9. Switching Catalysis from Hydrolysis to Perhydrolysis in Pseudomonas fluorescens Esterase

    Energy Technology Data Exchange (ETDEWEB)

    Yin, D.; Bernhardt, P; Morley, K; Jiang, Y; Cheeseman, J; Purpero, V; Schrag, J; Kazlauskas, R

    2010-01-01

    Many serine hydrolases catalyze perhydrolysis, the reversible formation of peracids from carboxylic acids and hydrogen peroxide. Recently, we showed that a single amino acid substitution in the alcohol binding pocket, L29P, in Pseudomonas fluorescens (SIK WI) aryl esterase (PFE) increased the specificity constant of PFE for peracetic acid formation >100-fold [Bernhardt et al. (2005) Angew. Chem., Int. Ed. 44, 2742]. In this paper, we extend this work to address the three following questions. First, what is the molecular basis of the increase in perhydrolysis activity? We previously proposed that the L29P substitution creates a hydrogen bond between the enzyme and hydrogen peroxide in the transition state. Here we report two X-ray structures of L29P PFE that support this proposal. Both structures show a main chain carbonyl oxygen closer to the active site serine as expected. One structure further shows acetate in the active site in an orientation consistent with reaction by an acyl-enzyme mechanism. We also detected an acyl-enzyme intermediate in the hydrolysis of {var_epsilon}-caprolactone by mass spectrometry. Second, can we further increase perhydrolysis activity? We discovered that the reverse reaction, hydrolysis of peracetic acid to acetic acid and hydrogen peroxide, occurs at nearly the diffusion limited rate. Since the reverse reaction cannot increase further, neither can the forward reaction. Consistent with this prediction, two variants with additional amino acid substitutions showed 2-fold higher k{sub cat}, but K{sub m} also increased so the specificity constant, k{sub cat}/K{sub m}, remained similar. Third, how does the L29P substitution change the esterase activity? Ester hydrolysis decreased for most esters (75-fold for ethyl acetate) but not for methyl esters. In contrast, L29P PFE catalyzed hydrolysis of {var_epsilon}-caprolactone five times more efficiently than wild-type PFE. Molecular modeling suggests that moving the carbonyl group closer to the

  10. Effects of gamma irradiation on the survival of Pseudomonas fluorescens inoculated on romaine lettuce

    Science.gov (United States)

    Irradiation of fresh fruits and vegetables has been utilized as a post-harvest intervention measure for inactivation of microbial pathogens. Pseudomonas fluorescens 2-79, Q8R1, Q287 are non-plant pathogenic, non-pectolytic strains of a potential biocontrol agent. We tested the sensitivity of these s...

  11. Sensitivity of Pseudomonas fluorescens to gamma irradiation following surface inoculations on romaine lettuce and baby spinach

    Science.gov (United States)

    Irradiation of fresh fruits and vegetables is a post-harvest intervention measure often used to inactivate pathogenic food-borne microbes. We evaluated the sensitivity of Pseudomonas fluorescens strains (2-79, Q8R1, Q287) to gamma irradiation following surface inoculations on romaine lettuce and spi...

  12. Evolutionary history of the phl gene cluster in the plant-associated bacterium Pseudomonas fluorescens

    NARCIS (Netherlands)

    Moynihan, J.A.; Morrissey, J.P.; Coppoolse, E.; Stiekema, W.J.; O'Gara, F.; Boyd, E.F.

    2009-01-01

    Pseudomonas fluorescens is of agricultural and economic importance as a biological control agent largely because of its plant-association and production of secondary metabolites, in particular 2, 4-diacetylphloroglucinol (2, 4-DAPG). This polyketide, which is encoded by the eight gene phl cluster,

  13. Improved Performance of Pseudomonas fluorescens lipase by covalent immobilization onto Amberzyme

    NARCIS (Netherlands)

    Aslan, Yakup; Handayani, Nurrahmi; Stavila, Erythrina; Loos, Katja

    2013-01-01

    Objective: In this study, the conditions of covalent immobilization of Pseudomonas fluorescens lipase onto an oxirane-activated support (Amberzyme) were optimized to obtain a high activity yield. Furthermore, the operational and storage stabilities of immobilized lipase were tested. Methods: Optimum

  14. Effect of GABA, a Bacterial Metabolite, on Pseudomonas fluorescens Surface Properties and Cytotoxicity

    Directory of Open Access Journals (Sweden)

    Marc G. J. Feuilloley

    2013-06-01

    Full Text Available Different bacterial species and, particularly Pseudomonas fluorescens, can produce gamma-aminobutyric acid (GABA and express GABA-binding proteins. In this study, we investigated the effect of GABA on the virulence and biofilm formation activity of different strains of P. fluorescens. Exposure of a psychotropic strain of P. fluorescens (MF37 to GABA (10−5 M increased its necrotic-like activity on eukaryotic (glial cells, but reduced its apoptotic effect. Conversely, muscimol and bicuculline, the selective agonist and antagonist of eukaryote GABAA receptors, respectively, were ineffective. P. fluorescens MF37 did not produce biosurfactants, and its caseinase, esterase, amylase, hemolytic activity or pyoverdine productions were unchanged. In contrast, the effect of GABA was associated to rearrangements of the lipopolysaccharide (LPS structure, particularly in the lipid A region. The surface hydrophobicity of MF37 was marginally modified, and GABA reduced its biofilm formation activity on PVC, but not on glass, although the initial adhesion was increased. Five other P. fluorescens strains were studied, and only one, MFP05, a strain isolated from human skin, showed structural differences of biofilm maturation after exposure to GABA. These results reveal that GABA can regulate the LPS structure and cytotoxicity of P. fluorescens, but that this property is specific to some strains.

  15. Phloroglucinol mediates crosstalk between the pyoluteorin and 2,4-diacetylphloroglucinol biosynthetic pathways in Pseudomonas fluorescens Pf-5

    Science.gov (United States)

    The antibiotics pyoluteorin and 2,4-diacetylphloroglucinol (DAPG) are involved in the biological control of certain soil-borne diseases by some strains of Pseudomonas fluorescens, including P. fluorescens Pf-5. These secondary metabolites also act as signaling molecules with each compound reported ...

  16. Iron-regulated metabolites of plant growth-promoting Pseudomonas fluorescens WCS374 : Their role in induced systemic resistance

    NARCIS (Netherlands)

    Djavaheri, M.

    2007-01-01

    The plant growth-promoting rhizobacterium Pseudomonas fluorescens WCS374r effectively suppresses fusarium wilt in radish by induced systemic resistance (ISR). In radish, WCS374r-mediated ISR depends partly on iron-regulated metabolites. Under iron-limiting conditions, P. fluorescens WCS374r produces

  17. Atmospheric-pressure air microplasma jets in aqueous media for the inactivation of Pseudomonas fluorescens cells

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Xianhui; Yang, Si-ze [Fujian Provincial Key Laboratory of Plasma and Magnetic Resonance, School of Physics and Mechanical and Electrical Engineering, Xiamen University, Xiamen, Fujian 361005 (China); Liu, Dongping [Fujian Provincial Key Laboratory of Plasma and Magnetic Resonance, School of Physics and Mechanical and Electrical Engineering, Xiamen University, Xiamen, Fujian 361005 (China); School of Physics and Materials Engineering, Dalian Nationalities University, Dalian 116600 (China); Song, Ying [School of Physics and Materials Engineering, Dalian Nationalities University, Dalian 116600 (China); School of Physics and Optoelectronic Technology, Dalian University of Technology, Dalian 116023 (China); Sun, Yue [School of Physics, Changchun University of Science and Technology, Changchun 130022 (China)

    2013-05-15

    The hollow fiber-based cold air microplasma jet array running at atmospheric pressure has been designed to inactivate Pseudomonas fluorescens (P. fluorescens) cells in vitro in aqueous media. The influences of electrode configurations, air flow rate, and applied voltage on the discharge characteristics of the single microplasma jet operating in aqueous media are presented, and the bactericidal efficiency of the hollow fibers-based and large-volume microplasma jet array is reported. Optical emission spectroscopy is utilized to identify excited species during the antibacterial testing of plasma in solutions. These well-aligned and rather stable air microplasma jets containing a variety of short-lived species, such as OH and O radicals and charged particles, are in direct contact with aqueous media and are very effective in killing P. fluorescens cells in aqueous media. This design shows its potential application for atmospheric pressure air plasma inactivation of bacteria cells in aqueous media.

  18. Getting the ecology into interactions between plants and the plant growth-promoting bacterium Pseudomonas fluorescens.

    Science.gov (United States)

    Hol, W H Gera; Bezemer, T Martijn; Biere, Arjen

    2013-01-01

    Plant growth-promoting rhizobacteria (PGPR) are increasingly appreciated for their contributions to primary productivity through promotion of growth and triggering of induced systemic resistance in plants. Here we focus on the beneficial effects of one particular species of PGPR (Pseudomonas fluorescens) on plants through induced plant defense. This model organism has provided much understanding of the underlying molecular mechanisms of PGPR-induced plant defense. However, this knowledge can only be appreciated at full value once we know to what extent these mechanisms also occur under more realistic, species-diverse conditions as are occurring in the plant rhizosphere. To provide the necessary ecological context, we review the literature to compare the effect of P. fluorescens on induced plant defense when it is present as a single species or in combination with other soil dwelling species. Specifically, we discuss combinations with other plant mutualists (bacterial or fungal), plant pathogens (bacterial or fungal), bacterivores (nematode or protozoa), and decomposers. Synergistic interactions between P. fluorescens and other plant mutualists are much more commonly reported than antagonistic interactions. Recent developments have enabled screenings of P. fluorescens genomes for defense traits and this could help with selection of strains with likely positive interactions on biocontrol. However, studies that examine the effects of multiple herbivores, pathogens, or herbivores and pathogens together on the effectiveness of PGPR to induce plant defenses are underrepresented and we are not aware of any study that has examined interactions between P. fluorescens and bacterivores or decomposers. As co-occurring soil organisms can enhance but also reduce the effectiveness of PGPR, a better understanding of the biotic factors modulating P. fluorescens-plant interactions will improve the effectiveness of introducing P. fluorescens to enhance plant production and defense.

  19. Biosynthesis and regulation of cyclic lipopeptides in Pseudomonas fluorescens

    NARCIS (Netherlands)

    Bruijn, de I.

    2009-01-01

    Cyclic lipopeptides (CLPs) are surfactant and antibiotic metabolites produced by a variety of bacterial
    genera. For the genus Pseudomonas, many structurally different CLPs have been identified. CLPs play an
    important role in surface motility of Pseudomonas strains, but also in virulence

  20. SANITATION PROCESS OPTIMALIZATION IN RELATION TO THE MICROBIAL BIOFILM OF PSEUDOMONAS FLUORESCENS

    Directory of Open Access Journals (Sweden)

    Vladimír Vietoris

    2012-02-01

    Full Text Available Biofilms have been of considerable interest in the context of food hygiene. Extracellular polymeric substances play an important role in the attachment and colonization of microorganisms to food-contact surfaces. If the microorganisms from food-contact surfaces are not completely removed, they may lead to biofilm formation and also increase the biotransfer potential. The experimental part was focused on the adhesion of bacterial cells under static conditions and testing the effectiveness of disinfectants on created biofilm. In laboratory conditions we prepared and formed the bacterial biofilms Pseudomonas fluorescens in the test surfaces of stainless steel. Over the 72 hours and the next 72 hours were observed numbers of adhesion bacterial cells of Pseudomonas fluorescens on solid surfaces of tested materials.

  1. Effect of Naphthalene and Salicylate Analogues on the Bioluminescence of Bioreporter Pseudomonas Fluorescens HK44.

    Czech Academy of Sciences Publication Activity Database

    Trögl, Josef; Kuncová, Gabriela; Kubicová, L.; Pařík, P.; Hálová, Jaroslava; Demnerová, K.; Ripp, S.; Sayler, G. S.

    2007-01-01

    Roč. 52, 1 (2007) , s. 3-14 ISSN 0015-5632 R&D Projects: GA ČR(CZ) GA104/05/2637; GA ČR(CZ) GA203/06/1244 Institutional research plan: CEZ:AV0Z40720504; CEZ:AV0Z40320502 Keywords : pseudomonas fluorescens HK44 * bioluminescence * bioluminescence Subject RIV: CE - Biochemistry Impact factor: 0.989, year: 2007

  2. Detection of Pseudomonas fluorescens from broth, water and ...

    African Journals Online (AJOL)

    sonal

    2015-04-08

    Apr 8, 2015 ... Author(s) agree that this article remains permanently open access under the terms of ... grown in nutrient broth overnight, pond water, mucus and kidney ... a rapid test for detection of Pseudomonas strains in milk is required.

  3. Control of Fusarium verticillioides, cause of ear rot of maize, by Pseudomonas fluorescens.

    Science.gov (United States)

    Nayaka, Siddaiah Chandra; Shankar, Arakere C Udaya; Reddy, Munagala S; Niranjana, Siddapura R; Prakash, Harishchandra S; Shetty, Hunthrike S; Mortensen, Carmen N

    2009-07-01

    Maize is one of the staple food crops grown in India. Fusarium verticillioides (Sacc.) Nirenberg is the most important fungal pathogen of maize, associated with diseases such as ear rot and kernel rot. Apart from the disease, it is capable of producing fumonisins, which have elicited considerable attention over the past decade owing to their association with animal disease syndromes. Hence, the present study was conducted to evaluate ecofriendly approaches by using a maize rhizosphere isolate of Pseudomonas fluorescens (Trev.) Mig. and its formulation to control ear rot disease and fumonisin accumulation, and also to study the capacity to promote growth and yield of maize. In vitro assays were conducted to test the efficacy of P. fluorescens as a seed treatment on seed germination, seedling vigour and also the incidence of F. verticillioides in different maize cultivars. The field trials included both seed treatment and foliar spray. For all the experiments, P. fluorescens was formulated using corn starch, wheat bran and talc powder. In each case there were three different treatments of P. fluorescens, a non-treated control and chemical control. Pure culture and the formulations, in comparison with the control, increased plant growth and vigour as measured by seed germination, seedling vigour, plant height, 1000 seed weight and yield. P. fluorescens pure culture used as seed treatment and as spray treatment enhanced the growth parameters and reduced the incidence of F. verticillioides and the level of fumonisins to a maximum extent compared with the other treatments. The study demonstrates the potential role of P. fluorescens and its formulations in ear rot disease management. The biocontrol potential of this isolate is more suited for fumonisin reduction in maize kernels intended for human and animal feed. (c) 2009 Society of Chemical Industry.

  4. Inhibition of Vibrio anguillarum by Pseudomonas fluorescens AH2, a possible probiotic treatment of fish

    DEFF Research Database (Denmark)

    Gram, Lone; Melchiorsen, Jette; Spanggaard, Bettina

    1999-01-01

    To study the possible use of probiotics in fish farming, we evaluated the in vitro and in vivo antagonism of antibacterial strain Pseudomonas fluorescens strain AH2 against the fish- pathogenic bacterium Vibrio anguillarum. As iron is important in virulence and bacterial interactions, the effect....... fluorescens AH2 inhibited the growth of V. anguillarum during coculture, independently of the iron concentration, when the initial count of the antagonist was 100 to 1,000 times greater that of the fish pathogen. These in vitro results were successfully repeated in vivo. A probiotic effect in vivo was tested...... by exposing rainbow trout (Oncorynchus mykiss Walbaum) to P. fluorescens AH2 at a density of 10(5) CFU/ml for 5 days before a challenge with V. anguillarum at 10(4) to 10(5) CFU/ml for 1 h. Some fish were also exposed to P. fluorescens AH2 at 10(7) CFU/ml during the 1-h infection. The combined probiotic...

  5. Interactions of trivalent and tetravalent heavy metal-siderophore complexes with pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Yoshida, T.; Ozaki, T.; Ohnuki, T.; Francis, A.J.

    2004-01-01

    We investigated the interactions of the Fe(III)-, Eu(III)-, and Hf(IV)-desferrioxamine B (DFO) complexes with the Gram-negative aerobic bacterium Pseudomonas fluorescens. Potentiometric titration of 1:1 Fe(III)-, Eu(III)-, and Hf(IV)-DFO complexes showed that Hf(IV) formed a strong complex with DFO whose stability was comparable to that of the Fe(III)-DFO complex, while Eu(III) formed a weaker one. DFO in a growth medium was not degraded by P. fluorescens. Contact of P. fluorescens cells with the Fe(III)-, Eu(III)-, and Hf(IV)-DFO complexes at pH 4-9 revealed that there was negligible adsorption of Hf(IV) and Fe(III), whereas Eu(III) was dissociated from DFO and was readily adsorbed by the cells. These results suggest that Fe(III) and Hf(IV) form stable complexes with DFO and are not adsorbed by P. fluorescens cells. Europium(III) forms a weaker complex with DFO than Fe(III) and Hf(IV) do and its DFO complex is readily dissociated in the presence of the cells. (orig.)

  6. Inactivation of Pseudomonas fluorescens in skim milk by combinations of pulsed electric fields and organic acids.

    Science.gov (United States)

    Fernández-Molina, Juan J; Altunakar, Bilge; Bermúdez-Aguirre, Daniela; Swanson, Barry G; Barbosa-Cánovas, Gustavo V

    2005-06-01

    Pseudomonas fluorescens suspended in skim milk was inactivated by application of pulsed electric fields (PEF) either alone or in combination with acetic or propionic acid. The initial concentration of microorganisms ranged from 10(5) to 10(6) CFU/ml. Addition of acetic acid and propionic acid to skim milk inactivated 0.24 and 0.48 log CFU/ml P. fluorescens, respectively. Sets of 10, 20, and 30 pulses were applied to the skim milk using exponentially decaying pulses with pulse lengths of 2 micros and pulse frequencies of 3 Hz. Treatment temperature was maintained between 16 and 20 degrees C. In the absence of organic acids, PEF treatment of skim milk at field intensities of 31 and 38 kV/cm reduced P. fluorescens populations by 1.0 to 1.8 and by 1.2 to 1.9 log CFU/ml, respectively. Additions of acetic and propionic acid to the skim milk in a pH range of 5.0 to 5.3 and PEF treatment at 31, 33, and 34 kV/cm, and 36, 37, and 38 kV/cm reduced the population of P. fluorescens by 1.4 and 1.8 log CFU/ml, respectively. No synergistic effect resulted from the combination of PEF with acetic or propionic acid.

  7. Production and properties of biosurfactants from a newly isolated Pseudomonas fluorescens HW-6 growing on hexandecane

    Energy Technology Data Exchange (ETDEWEB)

    Vasileva-Tonkova, E.; Galabova, D. [Bulgarian Academy of Sciences, Dept. of Microbial Biochemistry, Sofia (Bulgaria); Stoimenova, E.; Lalchev, Z. [Dept. of Biochemistry, Sofia Univ. ' ' St. Kliment Ohridski' ' , Sofia (Bulgaria)

    2006-07-15

    The newly isolated from industrial wastewater Pseudomonas fluorescens strain HW-6 produced glycolipid biosurfactants at high concentrations (1.4-2.0 g 1{sup -1}) when grown on hexadecane as a sole carbon source. Biosurfactants decreased the surface tension of the air/water interface by 35 mN m{sup -1} and possessed a low critical micelle concentration value of 20 mg 1{sup -1}, which indicated high surface activity. They efficiently emulsified aromatic hydrocarbons, kerosene, n-paraffins and mineral oils. Biosurfactant production contributed to a significant increase in cell hydrophobicity correlated with an increased growth of the strain on hexadecane. The results suggested that the newly isolated strain of Ps. fluorescens and produced glycolipid biosurfactants with effective surface and emulsifying properties are very promising and could find application for bioremediation of hydrocarbon-polluted sites. (orig.)

  8. Genetic Control of Plant Root Colonization by the Biocontrol agent, Pseudomonas fluorescens

    Energy Technology Data Exchange (ETDEWEB)

    Cole, Benjamin J.; Fletcher, Meghan; Waters, Jordan; Wetmore, Kelly; Blow, Matthew J.; Deutschbauer, Adam M.; Dangl, Jeffry L.; Visel, Axel

    2015-03-19

    Plant growth promoting rhizobacteria (PGPR) are a critical component of plant root ecosystems. PGPR promote plant growth by solubilizing inaccessible minerals, suppressing pathogenic microorganisms in the soil, and directly stimulating growth through hormone synthesis. Pseudomonas fluorescens is a well-established PGPR isolated from wheat roots that can also colonize the root system of the model plant, Arabidopsis thaliana. We have created barcoded transposon insertion mutant libraries suitable for genome-wide transposon-mediated mutagenesis followed by sequencing (TnSeq). These libraries consist of over 105 independent insertions, collectively providing loss-of-function mutants for nearly all genes in the P.fluorescens genome. Each insertion mutant can be unambiguously identified by a randomized 20 nucleotide sequence (barcode) engineered into the transposon sequence. We used these libraries in a gnotobiotic assay to examine the colonization ability of P.fluorescens on A.thaliana roots. Taking advantage of the ability to distinguish individual colonization events using barcode sequences, we assessed the timing and microbial concentration dependence of colonization of the rhizoplane niche. These data provide direct insight into the dynamics of plant root colonization in an in vivo system and define baseline parameters for the systematic identification of the bacterial genes and molecular pathways using TnSeq assays. Having determined parameters that facilitate potential colonization of roots by thousands of independent insertion mutants in a single assay, we are currently establishing a genome-wide functional map of genes required for root colonization in P.fluorescens. Importantly, the approach developed and optimized here for P.fluorescens>A.thaliana colonization will be applicable to a wide range of plant-microbe interactions, including biofuel feedstock plants and microbes known or hypothesized to impact on biofuel-relevant traits including biomass productivity

  9. Supervivencia de Pseudomonas fluorescens en suelos con diferente contenido de materia orgánica Pseudomonas fluorescens survival in soils with different contents of organic matter

    Directory of Open Access Journals (Sweden)

    E.B.R. Perotti

    2005-06-01

    Full Text Available Pseudomonas fluorescens es una bacteria PGPR (plant growth promoting rhizobacteria, heterótrofa, capaz de combatir fitopatógenos edáficos. Su supervivencia podría estar favorecida por el elevado contenido de materia orgánica del suelo (MOS. Para probarlo, se inocularon, en condiciones de laboratorio, tres cepas de P. fluorescens: UP61, C7R12, y P190 (nativa de Balcarce, Buenos Aires en suelos rizosféricos de tomate representativos de diferentes zonas de Argentina: suelo Argiudol (Balcarce, y Zavalla, Santa Fe y suelo Torrifluvens (Cipolletti, Río Negro (MOS %: 7,2; 4,3 y 2,6 respectivamente. Los resultados indicaron que la supervivencia de P. fluorescens en los suelos Argiudoles fue similar; aunque las pendientes de las curvas de supervivencia en el suelo de Zavalla fueron menores que las observadas en el suelo de Balcarce. La producción de CO2 fue superior en el suelo de Balcarce que en el suelo de Zavalla (4,3 y 2,8 mmol.g-1suelo, esta diferencia podría ser explicada por la existencia de una mayor presión competitiva por parte de la microflora nativa. La supervivencia en el suelo Torrifluvens resultó mínima, lo que sería atribuible a su elevada conductividad eléctrica más que al menor contenido de MOS. La cepa UP61 presentó en todos los casos la mejor supervivencia.Pseudomonas fluorescens are plant growth promoting rhizobacteria (PGPR. The survival of this inoculated heterotrophic bacterium may be affected by soil organic matter content (SOM. To confirm this hypothesis, three strains of P. fluorescens: UP61, C7R12 y P190 (native of Balcarce, Buenos Aires were inoculated, in laboratory conditions, into three argentine rhizospheric soils: two Argiudolls (Balcarce, and Zavalla, Santa Fe and a Torrifluvens (Cipolletti, Río Negro with different SOM: 7,2; 4,3; and 2,6%, respectibily. The results indicated that the all three isolates survival in general was not different. The slopes of the regression curves in Zavalla soil were very

  10. Exposure-related effects of Pseudomonas fluorescens (Pf-CL145A) on juvenile unionid mussels

    Science.gov (United States)

    Weber, Kerry L.; Luoma, James A.; Mayer, Denise A.; Aloisi, Douglas B.; Eckert, Nathan L.

    2015-01-01

    The exposure-related effects of a commercially prepared spray-dried powder (SDP) or freeze-dried powder (FDP) formulation of Pseudomonas fluorescens (strain CL145A) on the survival of seven species of newly metamorphosed (<72 hours old) freshwater unionid mussels was evaluated. Forty unionid mussels of each species were randomly distributed to test chambers and each species independently exposed for 24 hours to a static dose of either SDP (four species: Lampsilis cardium, Lampsilis siliquoidea, Lampsilis higginsii, andLigumia recta) or FDP (three species: Obovaria olivaria, Actinonaias ligamentina, andMegalonaias nervosa).

  11. The Influence of Pseudomonas fluorescens on Corrosion Products of Archaeological Tin-Bronze Analogues

    Science.gov (United States)

    Ghiara, G.; Grande, C.; Ferrando, S.; Piccardo, P.

    2018-01-01

    In this study, tin-bronze analogues of archaeological objects were investigated in the presence of an aerobic Pseudomonas fluorescens strain in a solution, containing chlorides, sulfates, carbonates and nitrates according to a previous archaeological characterization. Classical fixation protocols were employed in order to verify the attachment capacity of such bacteria. In addition, classical metallurgical analytical techniques were used to detect the effect of bacteria on the formation of uncommon corrosion products in such an environment. Results indicate quite a good attachment capacity of the bacteria to the metallic surface and the formation of the uncommon corrosion products sulfates and sulfides is probably connected to the bacterial metabolism.

  12. Sorption of Eu(III) on Pseudomonas fluorescens in the presence of citric acid

    International Nuclear Information System (INIS)

    Suzuki, Yoshinori; Tsushima, Satoru; Yamamoto, Ichiro; Nankawa, Takuya; Yoshida, Takahiro; Ozaki, Takuo; Ohnuki, Toshihiko; Francis, Arokiasamy J.; Enokida, Youichi

    2005-01-01

    We studied the sorption of Eu(III) on Pseudomonas fluorescens in the absence and presence of citric acid by a batch method. The cells were placed in a solution containing 2 μM of Eu(III) and 0, 100, or 1000 μM of citric acid at pH 3 9 for 5 hours. In the absence of citric acid, almost 100% of Eu(III) was sorbed on P. fluorescens at pHs below 7; above 7, sorption decreased with an increase in pH. The time course of Eu(III) sorption on P. fluorescens showed that a fraction of it was desorbed into the solution at alkaline pHs, suggesting that the bacterium may release some exudates. With citric acid present, we found that at higher concentrations there was lower sorption of Eu(III), reflecting the formation of Eu(III)-citrate complexes with the Eu(III)-cell-surface complexes. This decrease in Eu(III) sorption was significant in alkaline pHs. These findings suggest that citric acid which is ubiquitously found in the environment enhances migration of trivalent actinides in the alkaline environment. (author)

  13. Characterization of the SPI-1 and Rsp type three secretion systems in Pseudomonas fluorescens F113.

    Science.gov (United States)

    Barret, Matthieu; Egan, Frank; Moynihan, Jennifer; Morrissey, John P; Lesouhaitier, Olivier; O'Gara, Fergal

    2013-06-01

    Pseudomonas fluorescens F113 is a plant growth-promoting rhizobacterium (PGPR) isolated from the sugar beet rhizosphere. The recent annotation of the F113 genome sequence has revealed that this strain encodes a wide array of secretion systems, including two complete type three secretion systems (T3SSs) belonging to the Hrp1 and SPI-1 families. While Hrp1 T3SSs are frequently encoded in other P. fluorescens strains, the presence of a SPI-1 T3SS in a plant-beneficial bacterial strain was unexpected. In this work, the genetic organization and expression of these two T3SS loci have been analysed by a combination of transcriptional reporter fusions and transcriptome analyses. Overexpression of two transcriptional activators has shown a number of genes encoding putative T3 effectors. In addition, the influence of these two T3SSs during the interaction of P. fluorescens F113 with some bacterial predators was also assessed. Our data revealed that the transcriptional activator hilA is induced by amoeba and that the SPI-1 T3SS could potentially be involved in resistance to amoeboid grazing. © 2013 John Wiley & Sons Ltd and Society for Applied Microbiology.

  14. Metabolic functions of Pseudomonas fluorescens strains from Populus deltoides depend on rhizosphere or endosphere isolation compartment

    Directory of Open Access Journals (Sweden)

    Collin M Timm

    2015-10-01

    Full Text Available The bacterial microbiota of plants is diverse, with 1,000s of operational taxonomic units (OTUs associated with any individual plant. In this work we investigate the differences between 19 sequenced Pseudomonas fluorescens strains, isolated from Populus deltoides rhizosphere and endosphere and which represent a single OTU, using phenotypic analysis, comparative genomics, and metabolic models. While no traits were exclusive to either endosphere or rhizosphere P. fluorescens isolates, multiple pathways relevant for plant-bacterial interactions are enriched in endosphere isolate genomes. Further, growth phenotypes such as phosphate solubilization, protease activity, denitrification and root growth promotion are biased towards endosphere isolates. Endosphere isolates have significantly more metabolic pathways for plant signaling compounds and an increased metabolic range that includes utilization of energy rich nucleotides and sugars, consistent with endosphere colonization. Rhizosphere P. fluorescens have fewer pathways representative of plant-bacterial interactions but show metabolic bias towards chemical substrates often found in root exudates. This work reveals the diverse functions that may contribute to colonization of the endosphere by bacteria and are enriched among closely related isolates.

  15. Pseudomonas fluorescens-like bacteria from the stomach: a microbiological and molecular study.

    Science.gov (United States)

    Patel, Saurabh Kumar; Pratap, Chandra Bhan; Verma, Ajay Kumar; Jain, Ashok Kumar; Dixit, Vinod Kumar; Nath, Gopal

    2013-02-21

    To characterize oxidase- and urease-producing bacterial isolates, grown aerobically, that originated from antral biopsies of patients suffering from acid peptic diseases. A total of 258 antral biopsy specimens were subjected to isolation of bacteria followed by tests for oxidase and urease production, acid tolerance and aerobic growth. The selected isolates were further characterized by molecular techniques viz. amplifications for 16S rRNA using universal eubacterial and HSP60 gene specific primers. The amplicons were subjected to restriction analysis and partial sequencing. A phylogenetic tree was generated using unweighted pair group method with arithmetic mean (UPGMA) from evolutionary distance computed with bootstrap test of phylogeny. Assessment of acidity tolerance of bacteria isolated from antrum was performed using hydrochloric acid from 10(-7) mol/L to 10(-1) mol/L. Of the 258 antral biopsy specimens collected from patients, 179 (69.4%) were positive for urease production by rapid urease test and 31% (80/258) yielded typical Helicobacter pylori (H. pylori) after 5-7 d of incubation under a microaerophilic environment. A total of 240 (93%) antral biopsies yielded homogeneous semi-translucent and small colonies after overnight incubation. The partial 16S rRNA sequences revealed that the isolates had 99% similarity with Pseudomonas species. A phylogenetic tree on the basis of 16S rRNA sequences denoted that JQ927226 and JQ927227 were likely to be related to Pseudomonas fluorescens (P. fluorescens). On the basis of HSP60 sequences applied to the UPGMA phylogenetic tree, it was observed that isolated strains in an aerobic environment were likely to be P. fluorescens, and HSP60 sequences had more discriminatory potential rather than 16S rRNA sequences. Interestingly, this bacterium was acid tolerant for hours at low pH. Further, a total of 250 (96.9%) genomic DNA samples of 258 biopsy specimens and DNA from 240 bacterial isolates were positive for the 613 bp

  16. Characterization of the biocontrol activity of pseudomonas fluorescens strain X reveals novel genes regulated by glucose.

    Directory of Open Access Journals (Sweden)

    Gerasimos F Kremmydas

    Full Text Available Pseudomonas fluorescens strain X, a bacterial isolate from the rhizosphere of bean seedlings, has the ability to suppress damping-off caused by the oomycete Pythium ultimum. To determine the genes controlling the biocontrol activity of strain X, transposon mutagenesis, sequencing and complementation was performed. Results indicate that, biocontrol ability of this isolate is attributed to gcd gene encoding glucose dehydrogenase, genes encoding its co-enzyme pyrroloquinoline quinone (PQQ, and two genes (sup5 and sup6 which seem to be organized in a putative operon. This operon (named supX consists of five genes, one of which encodes a non-ribosomal peptide synthase. A unique binding site for a GntR-type transcriptional factor is localized upstream of the supX putative operon. Synteny comparison of the genes in supX revealed that they are common in the genus Pseudomonas, but with a low degree of similarity. supX shows high similarity only to the mangotoxin operon of Ps. syringae pv. syringae UMAF0158. Quantitative real-time PCR analysis indicated that transcription of supX is strongly reduced in the gcd and PQQ-minus mutants of Ps. fluorescens strain X. On the contrary, transcription of supX in the wild type is enhanced by glucose and transcription levels that appear to be higher during the stationary phase. Gcd, which uses PQQ as a cofactor, catalyses the oxidation of glucose to gluconic acid, which controls the activity of the GntR family of transcriptional factors. The genes in the supX putative operon have not been implicated before in the biocontrol of plant pathogens by pseudomonads. They are involved in the biosynthesis of an antimicrobial compound by Ps. fluorescens strain X and their transcription is controlled by glucose, possibly through the activity of a GntR-type transcriptional factor binding upstream of this putative operon.

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

    International Nuclear Information System (INIS)

    Moniot, Sebastien; Elias, Mikael; Kim, Donghyo; Scott, Ken; Chabriere, Eric

    2007-01-01

    Crystallization of DING protein from P. fluorescens is reported. A complete data set was collected to 1.43 Å resolution. 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 (P2 1 ), with typical unit-cell parameters a = 36.7, b = 123.7, c = 40.8 Å, α = 90, β = 116.7, γ = 90°. Preliminary crystallographic analysis reveals good diffraction quality for these crystals and a 1.43 Å resolution data set has been collected

  18. Survival of a Rifampicin-Resistant Pseudomonas fluorescens Strain in Nine Mollisols

    Directory of Open Access Journals (Sweden)

    Tami L. Stubbs

    2014-01-01

    Full Text Available Pseudomonas fluorescens strain D7 (P.f. D7 is a naturally occurring soil bacterium that shows promise as a biological herbicide to inhibit growth of annual grass weeds, including downy brome (Bromus tectorum L., in crop- and rangelands. Pseudomonas fluorescens strain D7rif (P.f. D7rif is a rifampicin-resistant strain of P.f. D7. One of the greatest obstacles to successful biological weed control is survival of the organism under field conditions. Nine soils in the taxonomic order of Mollisols, collected from downy brome-infested areas of the Western and Central United States, were inoculated with P.f. D7rif and incubated in the laboratory to determine the effects of soil type, soil properties, incubation temperature, and soil water potential on survival of P.f. D7rif over 63 days. Silt loam soils from Lind, Washington, and Moro, Oregon, sustained the highest P.f. D7rif populations, and recovery was the lowest from Pendleton, Oregon soil. Survival and recovery of P.f. D7rif varied with soil type and temperature but not with the two soil water potentials tested. After 63 days, P.f. D7rif was recovered at levels greater than log 5.5 colony forming units (CFU g−1 soil from five of the nine test soils, a level adequate to suppress downy brome under field or range conditions.

  19. Spatial distributions of Pseudomonas fluorescens colony variants in mixed-culture biofilms.

    Science.gov (United States)

    Workentine, Matthew L; Wang, Siyuan; Ceri, Howard; Turner, Raymond J

    2013-07-28

    The emergence of colony morphology variants in structured environments is being recognized as important to both niche specialization and stress tolerance. Pseudomonas fluorescens demonstrates diversity in both its natural environment, the rhizosphere, and in laboratory grown biofilms. Sub-populations of these variants within a biofilm have been suggested as important contributors to antimicrobial stress tolerance given their altered susceptibility to various agents. As such it is of interest to determine how these variants might be distributed in the biofilm environment. Here we present an analysis of the spatial distribution of Pseudomonas fluorescens colony morphology variants in mixed-culture biofilms with the wildtype phenotype. These findings reveal that two variant colony morphotypes demonstrate a significant growth advantage over the wildtype morphotype in the biofilm environment. The two variant morphotypes out-grew the wildtype across the entire biofilm and this occurred within 24 h and was maintained through to 96 h. This competitive advantage was not observed in homogeneous broth culture. The significant advantage that the variants demonstrate in biofilm colonization over the wildtype denotes the importance of this phenotype in structured environments.

  20. LETHALITY OF PSEUDOMONAS FLUORESCENS STRAIN CLO145A TO THE 2 ZEBRA MUSSEL SPECIES PRESENT IN NORTH AMERICA

    International Nuclear Information System (INIS)

    Molloy, Daniel P.

    2001-01-01

    These experiments indicated that bacterial strain CL0145A of Pseudomonas fluorescens is equally lethal to the 2 zebra mussel species present in North America, Dreissena polymorpha and Dreissena bugensis. Thus, this bacterial strain should be equally effective at killing zebra mussels in power plant pipes, irrespective of which species is present

  1. Characterization of toxin complex gene clusters and insect toxicity of bacteria representing four subgroups of Pseudomonas fluorescens

    Science.gov (United States)

    Ten strains representing four lineages of Pseudomonas (P. chlororaphis, P. corrugata, P. koreensis, and P. fluorescens subgroups) were evaluated for toxicity to the tobacco hornworm Manduca sexta and the fruit fly Drosophila melanogaster. The three strains within the P. chlororaphis subgroup exhibi...

  2. Assessment of DAPG-producing Pseudomonas fluorescens for management of Meloidogyne incognita and Fusarium oxysporum on watermelon

    Science.gov (United States)

    Pseudomonas fluorescens isolates Clinto 1R, Wayne 1R and Wood 1R, which produce the antibiotic 2,4-diacetylphloroglucinol (DAPG), can suppress soilborne diseases and promote plant growth. Consequently, these beneficial bacterial isolates were tested on watermelon plants for suppression of Meloidogy...

  3. Roles of Rhizoxin and 2,4-diacetylphloroglucinol in Suppression of Fusarium spp. by the Rhizobacterium Pseudomonas fluorescens Pf-5

    Science.gov (United States)

    Pseudomonas fluorescens strain Pf-5 is a rhizosphere bacterium that acts as a biocontrol agent of soilborne plant diseases and produces at least 10 different secondary metabolites, including several with antifungal properties. We derived site-directed mutants of Pf-5 with single and multiple mutatio...

  4. LETHALITY OF PSEUDOMONAS FLUORESCENS STRAIN CLO145A TO THE 2 ZEBRA MUSSEL SPECIES PRESENT IN NORTH AMERICA

    Energy Technology Data Exchange (ETDEWEB)

    Daniel P. Molloy

    2001-10-28

    These experiments indicated that bacterial strain CL0145A of Pseudomonas fluorescens is equally lethal to the 2 zebra mussel species present in North America, Dreissena polymorpha and Dreissena bugensis. Thus, this bacterial strain should be equally effective at killing zebra mussels in power plant pipes, irrespective of which species is present.

  5. Metabolic engineering of Pseudomonas fluorescens for the production of vanillin from ferulic acid.

    Science.gov (United States)

    Di Gioia, Diana; Luziatelli, Francesca; Negroni, Andrea; Ficca, Anna Grazia; Fava, Fabio; Ruzzi, Maurizio

    2011-12-20

    Vanillin is one of the most important flavors in the food industry and there is great interest in its production through biotechnological processes starting from natural substrates such as ferulic acid. Among bacteria, recombinant Escherichia coli strains are the most efficient vanillin producers, whereas Pseudomonas spp. strains, although possessing a broader metabolic versatility, rapidly metabolize various phenolic compounds including vanillin. In order to develop a robust Pseudomonas strain that can produce vanillin in high yields and at high productivity, the vanillin dehydrogenase (vdh)-encoding gene of Pseudomonas fluorescens BF13 strain was inactivated via targeted mutagenesis. The results demonstrated that engineered derivatives of strain BF13 accumulate vanillin if inactivation of vdh is associated with concurrent expression of structural genes for feruloyl-CoA synthetase (fcs) and hydratase/aldolase (ech) from a low-copy plasmid. The conversion of ferulic acid to vanillin was enhanced by optimization of growth conditions, growth phase and parameters of the bioconversion process. The developed strain produced up to 8.41 mM vanillin, which is the highest final titer of vanillin produced by a Pseudomonas strain to date and opens new perspectives in the use of bacterial biocatalysts for biotechnological production of vanillin from agro-industrial wastes which contain ferulic acid. Copyright © 2011 Elsevier B.V. All rights reserved.

  6. Physiological and biochemical changes in Matricaria chamomilla induced by Pseudomonas fluorescens and water deficit stress

    Directory of Open Access Journals (Sweden)

    Hamid MOHAMMADI

    2018-04-01

    Full Text Available Environmental stresses and rhizosphere microorganisms affect growth parameters and accumulation of active ingredients especially in plants with medicinal properties. The present study examined the effects of chamomile (Matricaria chamomilla L. seedling inoculation with Pseudomonas fluorescens PF-135 strain on its growth parameters, photosynthetic pigments, proline, malondialdehyde (MDA, and hydrogen peroxide (H2O2 content, and essential oil concentration at both regular watering and water deficit experiments. Based on the obtained results, water deficit stress reduced root dry mass, and flower fresh and dry mass as well. However, amount of H2O2 and MDA in root and shoot tissues were considerably lower in inoculated plants compared to non-inoculated ones under both normal watering and water deficit regimes. It indicates that lipid peroxidation and production of reactive oxygen species has been diminished in inoculated plants. Also, essential oil content in inoculated plants significantly increased compared with that of non-inoculated ones under water deficit stress condition. It can be concluded that P. fluorescens PF-135 strain has an outstanding potential to alleviate adverse effects of water deficit on plant growth, and hence can be used as an excellent PGPR in order to boost chamomile productivity especially under water deficit stress condition.

  7. Compatibility of Azospirillum brasilense and Pseudomonas fluorescens in growth promotion of groundnut ( Arachis hypogea L.).

    Science.gov (United States)

    Prasad, Andhare A; Babu, Subramanian

    2017-01-01

    We attempted to study the compatibility among plant beneficial bacteria in the culture level by growing them near in the nutrient agar plates. Among all the bacteria tested, Rhizobium was found to inhibit the growth of other bacteria. From the compatible group of PGPR, we have selected one biofertilizer (Azospirillum brasilense strain TNAU) and one biocontrol agent (Pseudomonas fluorescens strain PF1) for further studies in the pot culture. We have also developed a bioformulation which is talc powder based, for individual bacteria and mixed culture. This formulation was used as seed treatment, soil application, seedling root dip and foliar spray in groundnut crop in vitro germination conditions. A. brasilense was found to enhance the tap root growth and P. fluorescens, the lateral root growth. The other growth parameters like shoot growth, number of leaves were enhanced by the combination of both of the bacteria than their individual formulations. Among the method of application tested in our study, soil application was found to be the best in yielding better results of plant growth promotion.

  8. Compatibility of Azospirillum brasilense and Pseudomonas fluorescens in growth promotion of groundnut ( Arachis hypogea L.

    Directory of Open Access Journals (Sweden)

    ANDHARE A. PRASAD

    Full Text Available ABSTRACT We attempted to study the compatibility among plant beneficial bacteria in the culture level by growing them near in the nutrient agar plates. Among all the bacteria tested, Rhizobium was found to inhibit the growth of other bacteria. From the compatible group of PGPR, we have selected one biofertilizer (Azospirillum brasilense strain TNAU and one biocontrol agent (Pseudomonas fluorescens strain PF1 for further studies in the pot culture. We have also developed a bioformulation which is talc powder based, for individual bacteria and mixed culture. This formulation was used as seed treatment, soil application, seedling root dip and foliar spray in groundnut crop in vitro germination conditions. A. brasilense was found to enhance the tap root growth and P. fluorescens, the lateral root growth. The other growth parameters like shoot growth, number of leaves were enhanced by the combination of both of the bacteria than their individual formulations. Among the method of application tested in our study, soil application was found to be the best in yielding better results of plant growth promotion.

  9. Whole-genome sequence of Pseudomonas fluorescens EK007-RG4, a promising biocontrol agent against a broad range of bacteria, including the fire blight bacterium Erwinia amylovora

    DEFF Research Database (Denmark)

    Habibi, Roghayeh; Tarighi, Saeed; Behravan, Javad

    2017-01-01

    Here, we report the first draft whole-genome sequence of Pseudomonas fluorescens strain EK007-RG4, which was isolated from the phylloplane of a pear tree. P. fluorescens EK007-RG4 displays strong antagonism against Erwinia amylovora, the causal agent for fire blight disease, in addition to several...

  10. Exposure-related effects of formulated Pseudomonas fluorescens strain CL145A to glochidia from seven unionid mussel species

    Science.gov (United States)

    Luoma, James A.; Weber, Kerry L.; Severson, Todd J.; Schreier, Theresa M.; Mayer, Denise A.; Aloisi, Douglas B.; Eckert, Nathan L.

    2015-01-01

    The study was completed to evaluate the exposure-related effects of a biopesticide for dreissenid mussel (Dreissena polymorpha, zebra mussel and Dreissena rostriformis bugensis, quagga mussel) control on glochidia from unionid mussels endemic to the Great Lakes and Upper Mississippi River Basins. The commercially prepared biopesticide was either a spray-dried powder (SDP) or freeze-dried powder (FDP) formulation of Pseudomonas fluorescens, strain CL145A. Glochidia of the unionid mussel species Lampsilis cardium, Lampsilis siliquoidea,Lampsilis higginsii, Ligumia recta, Obovaria olivaria, and Actinonaias ligamentina were exposed to SDP-formulated P. fluorescens andLampsilis cardium and Megalonaias nervosa were exposed to FDP-formulated P. fluorescens.

  11. Pseudomonas fluorescens strain CL145A - a biopesticide for the control of zebra and quagga mussels (Bivalvia: Dreissenidae).

    Science.gov (United States)

    Molloy, Daniel P; Mayer, Denise A; Gaylo, Michael J; Morse, John T; Presti, Kathleen T; Sawyko, Paul M; Karatayev, Alexander Y; Burlakova, Lyubov E; Laruelle, Franck; Nishikawa, Kimi C; Griffin, Barbara H

    2013-05-01

    Zebra mussels (Dreissena polymorpha) and quagga mussels (Dreissena rostriformis bugensis) are the "poster children" of high-impact aquatic invasive species. In an effort to develop an effective and environmentally acceptable method to control their fouling of raw-water conduits, we have investigated the potential use of bacteria and their natural metabolic products as selective biological control agents. An outcome of this effort was the discovery of Pseudomonas fluorescens strain CL145A - an environmental isolate that kills these dreissenid mussels by intoxication (i.e., not infection). In the present paper, we use molecular methods to reconfirm that CL145A is a strain of the species P. fluorescens, and provide a phylogenetic analysis of the strain in relation to other Pseudomonas spp. We also provide evidence that the natural product lethal to dreissenids is associated with the cell wall of P. fluorescens CL145A, is a heat-labile secondary metabolite, and has degradable toxicity within 24 h when applied to water. CL145A appears to be an unusual strain of P. fluorescens since it was the only one among the ten strains tested to cause high mussel mortality. Pipe trials conducted under once-through conditions indicated: (1) P. fluorescens CL145A cells were efficacious against both zebra and quagga mussels, with high mortalities achieved against both species, and (2) as long as the total quantity of bacterial cells applied during the entire treatment period was the same, similar mussel mortality could be achieved in treatments lasting 1.5-12.0 h, with longer treatment durations achieving lower mortalities. The efficacy data presented herein, in combination with prior demonstration of its low risk of non-target impact, indicate that P. fluorescens CL145A cells have significant promise as an effective and environmentally safe control agent against these invasive mussels. Copyright © 2012 Elsevier Inc. All rights reserved.

  12. Evaluación de la biorremediación en suelos contaminados con hidrocarburos utilizando pseudomonas fluorescens.

    OpenAIRE

    Pérez Pozo, Marco Rafael

    2018-01-01

    Since the discovery of hydrocarbons, their use in industries has caused severe environmental impacts, which have been occurring frequently in various parts of the Ecuadorian Amazon. In the present investigation the biodegradation capacity of Pseudomonas fluorescens was assessed by bioaumentation in soils of the Ecuadorian Amazon contaminated with PAHs (Polycyclic Aromatic Hydrocarbons) and TPHs (Total Petroleum Hydrocarbons). The bioaumentation, is an efficient alternative for this type of...

  13. Efficacy of Pseudomonas fluorescens (Pf-CL145A) spray dried powder for controlling zebra mussels adhering to test substrates

    Science.gov (United States)

    Luoma, James A.; Severson, Todd J.; Weber, Kerry L.; Mayer, Denise A.

    2015-01-01

    A mobile bioassay trailer was used to assess the efficacy of Pseudomonas fluorescens (Pf-CL145A) spray dried powder (SDP) formulation for controlling zebra mussels (Dreissena polymorpha) from two midwestern lakes: Lake Carlos (Alexandria, Minnesota) and Shawano Lake (Shawano, Wisconsin). The effects of SDP exposure concentration and exposure duration on zebra mussel survival were evaluated along with the evaluation of a benthic injection application technique to reduce the amount of SDP required to induce zebra mortality.

  14. Differential impact of some Aspergillus species on Meloidogyne javanica biocontrol by Pseudomonas fluorescens strain CHA0.

    Science.gov (United States)

    Siddiqui, I A; Shaukat, S S; Khan, A

    2004-01-01

    The aim was to determine the influence of some Aspergillus species on the production of nematicidal agent(s) in vitro and biocontrol of Meloidogyne javanica in tomato by Pseudomonas fluorescens strains CHA0 and CHA0/pME3424. Six species of Aspergillus, isolated from the rhizosphere of certain crops, produced a variety of secondary metabolites in vitro. Culture filtrate (CF) obtained from Ps. fluorescens strain CHA0 and its2,4-diacetylphloroglucinol overproducing mutant CHA0/pME3424 grown in King's B liquid medium caused significant mortality of M. javanica juveniles in vitro. Bacterial growth medium amended with CF of A. niger enhanced nematicidal and beta-galactosidase activities of fluorescent pseudomonads while A. quadrilineatus repressed such activities. Methanol or ethyl acetate extracts of the CF of A. niger markedly optimized bacterial efficacy to cause nematode deaths while hexane extract of the fungus had no influence on the nematicidal activity of the bacterial strains. A. niger applied alone or in conjunction with the bacterial inoculants inhibited root-knot nematode galling in tomato. On the other hand, A. quadrilineatus used alone or together with CHA0 did not inhibit nematode galling but when used in combination with strain CHA0/pME3424 did reduce galling intensity. Aspergillus niger enhances the production of nematicidal compounds by Ps. fluorescensin vitro and improves biocontrol potential of the bacterial inoculants in tomato while A. quadrilineatus reduces bacterial performance to suppress root-knot nematodes. Rhizosphere harbours a variety of micro-organisms including bacteria, fungi and viruses. Aspergillus species are ubiquitous in most agricultural soils and generally produce a variety of secondary metabolites. Such metabolites synthesized by Aspergillus species may influence the production of nematicidal agents and subsequent biocontrol performance of the bacterial inoculants against plant-parasitic nematodes. This fact needs to be taken into

  15. INFLUÊNCIA DA ATIVIDADE ENZIMÁTICA DE PSEUDOMONAS FLUORESCENS 041 EM LABNEH

    Directory of Open Access Journals (Sweden)

    Andreza Angélica Ferreira

    2012-04-01

    Full Text Available A refrigeração do leite proporciona a seleção de bactérias psicrotróficas deteriorantes produtoras de enzimas termoresistentes que podem comprometer a qualidade do leite e derivados. O objetivo deste trabalho foi avaliar as implicações causadas pela atividade enzimática de Pseudomonas fluorescens 041 na produção de Labneh. O leite pasteurizado foi inoculado intencionalmente com, aproximadamente, 106 UFC.mL-1 de P. fluorescens 041 e Labneh foi produzido imediatamente após inoculação no leite (tempo 0 e após 48, 72 e 96 horas de inoculação e armazenamento a 4 ºC. A qualidade físico-química do leite, do Labneh e do soro resultante da fabricação foi determinada. O rendimento prático, o rendimento técnico ajustado e o aproveitamento final de sólidos no Labneh em relação ao volume de leite (coeficiente GL também foram avaliados. Constatou-se alterações nas características físico-químicas do soro e do Labneh fabricado com leite armazenado a partir de 48 horas. Também, observou-se um aumento significativo em litros de leite destinado à produção de um quilo do produto ao longo do tempo de estocagem do leite inoculado com P. fluorescens 041, sendo os rendimentos prático, técnico ajustado e o coeficiente GL afetados pelo fator tempo. Portanto, a redução do tempo de estocagem do leite sob refrigeração e a prevenção da contaminação da matéria-prima por meio da adoção de boas práticas na cadeia do leite são medidas a serem adotadas para assegurar a qualidade e o rendimento do produto final.

  16. Impact du séchage sur la viabilité de Pseudomonas fluorescens (synthèse bibliographique

    Directory of Open Access Journals (Sweden)

    Mputu Kanyinda, JN.

    2014-01-01

    Full Text Available Impact of drying on Pseudomonas fluorescens viability. A review. Drying Pseudomonas fluorescens makes for more economical storage, transportation and marketing. The aim of the drying process is to stop and to stabilize all biological activity for optimal storage, compatible with the conservation of the maximum desired viability of the microorganisms. However, the viability rate of the bacteria after drying depends on the operating conditions of the drying process. One of the most important criteria to consider during the drying of biologically active products is the quality of the final dried product. Freeze-drying is the drying method most commonly used for Pseudomonas fluorescens. After their production, the bacteria are harvested by centrifugation and are freeze-dried, but the changes in temperature induced by freeze-drying are not without consequence for the cells. The freeze-drying process induces cell damage: peroxidation of fatty acids and proteins and DNA oxidation. However, use of protective compounds during freeze-drying and during storage increases significantly the rate of cell viability.

  17. Semi-scale production of PHAs from waste frying oil by Pseudomonas fluorescens S48

    Directory of Open Access Journals (Sweden)

    Rawia F. Gamal

    2013-01-01

    Full Text Available The present study aimed at developing a strategy to improve the volumetric production of PHAs by Pseudomonas fluorescens S48 using waste frying oil (WFO as the sole carbon source. For this purpose, several cultivations were set up to steadily improve nutrients supply to attain high cell density and high biopolymer productivity. The production of PHAs was examined in a 14 L bioreactor as one-stage batch, two-stage batch, and high-cell-density fed-batch cultures. The highest value of polymer content in one-stage bioreactor was obtained after 60 h (33.7%. Whereas, the two-stage batch culture increased the polymer content to 50.1% after 54 h. High-cell-density (0.64 g/L at continuous feeding rate 0.55 mL/l/h of WFO recorded the highest polymer content after 54 h (55.34%. Semi-scale application (10 L working volume increased the polymer content in one-stage batch, two-stage batch and high cell density fed-batch cultures by about 12.3%, 5.8% and 11.3%, respectively, as compared with that obtained in 2 L fermentation culture. Six different methods for biopolymer extraction were done to investigate their efficiency for optimum polymer recovery. The maximum efficiency of solvent recovery of PHA was attained by chloroform-hypochlorite dispersion extraction. Gas chromatography (GC analysis of biopolymer produced by Pseudomonas fluorescens S48 indicated that it solely composed of 3-hydrobutyric acid (98.7%. A bioplastic film was prepared from the obtained PHB. The isolate studied shares the same identical sequence, which is nearly the complete 16S rRNA gene. The identity of this sequence to the closest pseudomonads strains is about 98-99%. It was probably closely related to support another meaningful parsiomony analysis and construction of a phylogenetic tree. The isolate is so close to Egyptian strain named EG 639838.

  18. Heat Production by the Denitrifying Bacterium Pseudomonas fluorescens and the Dissimilatory Ammonium-Producing Bacterium Pseudomonas putrefaciens during Anaerobic Growth with Nitrate as the Electron Acceptor

    OpenAIRE

    Samuelsson, M.-O.; Cadez, P.; Gustafsson, L.

    1988-01-01

    The heat production rate and the simultaneous nitrate consumption and production and consumption of nitrite and nitrous oxide were monitored during the anaerobic growth of two types of dissimilatory nitrate reducers. Pseudomonas fluorescens, a denitrifier, consumed nitrate and accumulated small amounts of nitrite or nitrous oxide. The heat production rate increased steadily during the course of nitrate consumption and decreased rapidly concomitant with the depletion of the electron acceptors....

  19. Detoxification of selenite and mercury by reduction and mutual protection in the assimilation of both elements by Pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Belzile, Nelson; Wu Gaojun; Chen, Yu-Wei; Appanna, Vasu D.

    2006-01-01

    A study on the assimilation and detoxification of selenium and mercury and on the interaction between these two elements was conducted on Pseudomonas fluorescens. P. fluorescens was able to convert separately both elements to their elemental forms, which are less toxic and biologically less available. To study the converting mechanism of selenite to elemental Se, cells were grown in the presence of various selenite concentrations and several parameters such as extracellular protein concentrations, pH, carbohydrate concentrations, isocitrate dehydrogenase (ICDH) and malic enzyme were monitored. Transmission electron microscopy (TEM) and various analytical methods were applied to confirm the interaction between selenium and cell. The former appeared as a red precipitate localized predominantly in the consumed culture medium. P. fluorescens also resisted to the toxic effect of mercury by converting Hg 2+ to the volatile and less toxic form Hg . Mercury reductase was likely responsible for the conversion of Hg 2+ to Hg . More importantly, the interaction between mercury and selenium was also studied. The presence of selenite significantly reduced the accumulation of mercury in P. fluorescens. It was also interesting to note that mercury appeared to behave as a protecting agent against selenium intoxication as the bioaccumulation of Se was also inhibited by this metal. The formation of Se-Hg complexes could explain this mutual protective effect. No precipitate of elemental Se could be detected when Hg was present in the cultures

  20. Antimicrobial activity of four essential oils against pigmenting Pseudomonas fluorescens and biofilmproducing Staphylococcus aureus of dairy origin

    Directory of Open Access Journals (Sweden)

    Francesca Pedonese

    2017-12-01

    Full Text Available Essential oils (EOs are mixtures of secondary metabolites of plant origin with many useful properties, among which the antimicrobial activity is also of interest for the food industry. EOs can exert their antimicrobial potential both directly, in food products and active packaging, and indirectly, as sanitizing and anti-biofilm agents of food facility surfaces. Aim of this research was to evaluate the antimicrobial activity of four EOs (bergamot, cinnamon, manuka and thyme against Pseudomonas fluorescens and Staphylococcus aureus isolated from milk and dairy products. The chemical composition of EOs was evaluated by Gas Chromatography-Mass Spectrometry analysis. Minimum Inhibitory Concentration values were determined by a microplate method against 9 Ps. fluorescens from marketed mozzarella with blue discoloration defect, and 3 biofilm-producing S. aureus from milk. Reference ATCC strains were included. Pigment production activity by Ps. fluorescens was assessed both in culture and in cheese. EOs of manuka (leptospermone 23% and thyme (carvacrol 30%, pcymene 20%, thymol 15% showed the highest antimicrobial activity against S. aureus, MIC values were 0.012%-0.024% and 0.024% v/v, respectively; meanwhile EOs from thyme and cinnamon (cinnamaldehyde 55% exhibited the best activity against Ps. fluorescens with MIC values of 0.098%-0.195% and 0.195%-0.391% v/v, respectively. The antimicrobial activity of these EOs is promising and they could be exploited in the dairy production chain.

  1. Understanding the molecular basis of plant growth promotional effect of Pseudomonas fluorescens on rice through protein profiling.

    Science.gov (United States)

    Kandasamy, Saveetha; Loganathan, Karthiba; Muthuraj, Raveendran; Duraisamy, Saravanakumar; Seetharaman, Suresh; Thiruvengadam, Raguchander; Ponnusamy, Balasubramanian; Ramasamy, Samiyappan

    2009-12-24

    Plant Growth Promoting Rhizobacteria (PGPR), Pseudomonas fluorescens strain KH-1 was found to exhibit plant growth promotional activity in rice under both in-vitro and in-vivo conditions. But the mechanism underlying such promotional activity of P. fluorescens is not yet understood clearly. In this study, efforts were made to elucidate the molecular responses of rice plants to P. fluorescens treatment through protein profiling. Two-dimensional polyacrylamide gel electrophoresis strategy was adopted to identify the PGPR responsive proteins and the differentially expressed proteins were analyzed by mass spectrometry. Priming of P. fluorescens, 23 different proteins found to be differentially expressed in rice leaf sheaths and MS analysis revealed the differential expression of some important proteins namely putative p23 co-chaperone, Thioredoxin h- rice, Ribulose-bisphosphate carboxylase large chain precursor, Nucleotide diPhosphate kinase, Proteosome sub unit protein and putative glutathione S-transferase protein. Functional analyses of the differential proteins were reported to be directly or indirectly involved in growth promotion in plants. Thus, this study confirms the primary role of PGPR strain KH-1 in rice plant growth promotion.

  2. Understanding the molecular basis of plant growth promotional effect of Pseudomonas fluorescens on rice through protein profiling

    Directory of Open Access Journals (Sweden)

    Thiruvengadam Raguchander

    2009-12-01

    Full Text Available Abstract Background Plant Growth Promoting Rhizobacteria (PGPR, Pseudomonas fluorescens strain KH-1 was found to exhibit plant growth promotional activity in rice under both in-vitro and in-vivo conditions. But the mechanism underlying such promotional activity of P. fluorescens is not yet understood clearly. In this study, efforts were made to elucidate the molecular responses of rice plants to P. fluorescens treatment through protein profiling. Two-dimensional polyacrylamide gel electrophoresis strategy was adopted to identify the PGPR responsive proteins and the differentially expressed proteins were analyzed by mass spectrometry. Results Priming of P. fluorescens, 23 different proteins found to be differentially expressed in rice leaf sheaths and MS analysis revealed the differential expression of some important proteins namely putative p23 co-chaperone, Thioredoxin h- rice, Ribulose-bisphosphate carboxylase large chain precursor, Nucleotide diPhosphate kinase, Proteosome sub unit protein and putative glutathione S-transferase protein. Conclusion Functional analyses of the differential proteins were reported to be directly or indirectly involved in growth promotion in plants. Thus, this study confirms the primary role of PGPR strain KH-1 in rice plant growth promotion.

  3. Immobilization of Pseudomonas fluorescens lipase onto magnetic nanoparticles for resolution of 2-octanol.

    Science.gov (United States)

    Xun, Er-na; Lv, Xiao-li; Kang, Wei; Wang, Jia-xin; Zhang, Hong; Wang, Lei; Wang, Zhi

    2012-10-01

    The lipase from Pseudomonas fluorescens (Lipase AK, AKL) was immobilized onto the magnetic Fe(3)O(4) nanoparticles via hydrophobic interaction. Enzyme loading and immobilization yield were determined as 21.4±0.5 mg/g and 49.2±1.8 %, respectively. The immobilized AKL was successfully used for resolution of 2-octanol with vinyl acetate used as acyl donor. Effects of organic solvent, water activity, substrate ratio, and temperature were investigated. Under the optimum conditions, the preferred isomer for AKL is the (R)-2-octanol and the highest enantioselectivity (E=71.5±2.2) was obtained with a higher enzyme activity (0.197±0.01 μmol/mg/min). The results also showed that the immobilized lipase could be easily separated from reaction media by the magnetic steel and remained 89 % of its initial activity as well as the nearly unchanged enantioselectivity after five consecutive cycles, indicating a high stability in practical operation.

  4. Pit formation on stainless steel surfaces pre-treated with biosurfactants produced by Pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Dagbert, Catherine; Meylheuc, Thierry; Bellon-Fontaine, Marie-Noelle

    2008-01-01

    Today, it is widely established that the surface tension of water can be reduced by some microorganisms capable of synthesizing surface-active compounds called biosurfactants (BS). BS characteristics depend on the microorganism that produces them and therefore, on the microorganism culture conditions. Some studies on chemical surfactants have shown that the adsorption of surface-active compounds plays a major role in corrosion; indeed they are used as a good corrosion inhibition tool. The purpose of this study was first, to estimate the importance and behavior of the stainless steels passive film on the adsorption of BS, produced by the Gram negative bacteria Pseudomonas fluorescens, and secondly, to study the impact of these treatments on the pitting corrosion. In this paper, the galvanostatic polarization technique, used as accelerated method for determining the characteristic pit potentials on stainless steels, is examined. Pit growth, shape and cover formation were also observed. The surface topography of the corroded specimens was investigated using field emission scanning electron microscopy (FESEM)

  5. Pit formation on stainless steel surfaces pre-treated with biosurfactants produced by Pseudomonas fluorescens

    Energy Technology Data Exchange (ETDEWEB)

    Dagbert, Catherine [ECP-LGPM, Grande Voie des Vignes, 92295 Chatenay-Malabry (France)], E-mail: catherine.dagbert@ecp.fr; Meylheuc, Thierry; Bellon-Fontaine, Marie-Noelle [INRA, UMR 763 Bioadhesion et Hygiene des Materiaux, F-91300 Massy (France); AGROPARISTECH, UMR 763 Bioadhesion et Hygiene des Materiaux, F-91300 Massy (France)

    2008-12-01

    Today, it is widely established that the surface tension of water can be reduced by some microorganisms capable of synthesizing surface-active compounds called biosurfactants (BS). BS characteristics depend on the microorganism that produces them and therefore, on the microorganism culture conditions. Some studies on chemical surfactants have shown that the adsorption of surface-active compounds plays a major role in corrosion; indeed they are used as a good corrosion inhibition tool. The purpose of this study was first, to estimate the importance and behavior of the stainless steels passive film on the adsorption of BS, produced by the Gram negative bacteria Pseudomonas fluorescens, and secondly, to study the impact of these treatments on the pitting corrosion. In this paper, the galvanostatic polarization technique, used as accelerated method for determining the characteristic pit potentials on stainless steels, is examined. Pit growth, shape and cover formation were also observed. The surface topography of the corroded specimens was investigated using field emission scanning electron microscopy (FESEM)

  6. Effects of high intensity pulsed electric field and thermal treatments on a lipase from Pseudomonas fluorescens.

    Science.gov (United States)

    Bendicho, S; Estela, C; Giner, J; Barbosa-Cánovas, G V; Martin, O

    2002-01-01

    Milk and dairy products may contain microorganisms capable of secreting lipases that cause sensory defects and technological problems in the dairy industry. In this study, the effects of thermal and high-intensity pulsed electric field (HIPEF) treatments on an extracellular lipase from Pseudomonas fluorescens, suspended in a simulated skim milk ultrafiltrate (SMUF) have been evaluated. Heat treatments applied were up to 30 min from 50 to 90 degrees C. HIPEF treatments were carried out using pilot plant facilities in a batch or continuous flow mode, where treatment chambers consisted of parallel and coaxial configuration, respectively. Samples were subjected to up to 80 pulses at electric field intensities ranging from 16.4 to 37.3 kV/cm. This resulted in a lipase that was quite resistant to heat and also to HIPEF. High (75 degrees C-15 s) and low pasteurization treatments (63 degrees C-30 min) led to inactivations of 5 and 20%, respectively. Using the batch-mode HIPEF equipment, a 62.1% maximum activity depletion was achieved after 80 pulses at 27.4 kV/cm. However, when HIPEF treatments were applied in the continuous flow mode, an inactivation rate of just 13% was achieved, after applying 80 pulses at 37.3 kV/cm and 3.5 Hz. The results of both heat and HIPEF treatments on enzyme inactivation were adjusted with good agreement to a first-order kinetic model (R2 > 62.3%).

  7. Modeling high-intensity pulsed electric field inactivation of a lipase from Pseudomonas fluorescens.

    Science.gov (United States)

    Soliva-Fortuny, R; Bendicho-Porta, S; Martín-Belloso, O

    2006-11-01

    The inactivation kinetics of a lipase from Pseudomonas fluorescens (EC 3.1.1.3.) were studied in a simulated skim milk ultrafiltrate treated with high-intensity pulsed electric fields. Samples were subjected to electric field intensities ranging from 16.4 to 27.4 kV/cm for up to 314.5 micros, thus achieving a maximum inactivation of 62.1%. The suitability of describing experimental data using mechanistic first-order kinetics and an empirical model based on the Weibull distribution function is discussed. In addition, different mathematical expressions relating the residual activity values to field strength and treatment time are supplied. A first-order fractional conversion model predicted residual activity with good accuracy (A(f) = 1.018). A mechanistic insight of the model kinetics was that experimental values were the consequence of different structural organizations of the enzyme, with uneven resistance to the pulsed electric field treatments. The Weibull model was also useful in predicting the energy density necessary to achieve lipase inactivation.

  8. Combinatorial efficacy of Trichoderma spp. and Pseudomonas fluorescens to enhance suppression of cell wall degrading enzymes produced by Fusarium wilt of Arachis hypogaea.L

    Directory of Open Access Journals (Sweden)

    P Rajeswari

    2017-12-01

    Full Text Available Fusarium oxysporum, the soil borne pathogen causes vascular wilt, on majority of crop plants. It has been demonstrated that two different species of Trichoderma and Pseudomonas fluorescens suppress disease by different mechanisms. Therefore, application of a mixture of these biocontrol agents, and thus of several suppressive mechanisms, may represent a viable control strategy. A necessity for biocontrol by combinations of biocontrol agents can be the compatibility of the co-inoculated micro-organisms. Hence, compatibility between Trichoderma spp. and Pseudomonas fluorescens that have the ability to suppress Fusarium oxysporum in vitro on the activity of pectinolytic enzymes of Fusarium oxysporum. The activity of pectinolytic enzymes, i.e. pectin methyl esterase, endo and exo polymethylgalacturonases and exo and endo pectin trans eliminases produced by Fusarium oxysporum (Control was higher. Maximum inhibition of pectin methylesterase, exo and endo polymethylgalacturonase and exo and endopectin trans eliminase was shown by culture filtrate of Trichoderma viride + Pseudomonas fluorescens (Tv+Pf (1+2%, followed by Trichoderma harzianum + Pseudomonas fluorescens, (Th +Pf (1.5+2% and Trichoderma viride + Trichoderma harzianum (Tv+Th (1+1.5%. However, pathogenecity suppression of Fusarium oxysporum, a causative of Arachis hypogaea. L by the compatible combination of Trichodema viride + Pseudomonas fluorescens (1+2% was significantly better as compared to the single bio-agent. This indicates that specific interactions between biocontrol agents influence suppression of pathogenicity factors directly by combinations of these compatible bio-agents.

  9. Effect of Eu(III) on the degradation of malic acid by Pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Nankawa, T.; Ozaki, T.; Ohnuki, T.; Suzuki, Y.; Francis, A.J.

    2005-01-01

    Full text of publication follows: The transuranic elements, such as Am(III) and Cm(III), are highly toxic because they emit high-energy α particles and have long half-lives. To estimate their long-term environmental behavior, we need to elucidate degradation of actinide-organic complexes by microorganisms. We studied the biodegradation of Eu(III)-malic acid complexes by Pseudomonas fluorescens. Malic acid is ubiquitous in the environment and is one of the microbial metabolites that is part of the tri-carboxylic acid (TCA) cycle. Europium(III) is a good analogue for Am(III) and Cm(III). To investigate the effect of Eu(III) on the degradation of malic acid by P. fluorescens, we compared the degradation behavior of Eu(III)-malic acid complexes to that of Fe(III) and Al(III)-malic acid complexes. In the medium containing 1 mM malic acid and 0-0.5 mM Fe(III), malic acid was degraded completely. In the medium containing 1 mM malic acid and 0.05-0.5 mM Al(III), malic acid was degraded until the concentration of malic acid became equal to that of Al(III), indicating that Al(III)-malic acid complex with 1: 1 molar ratio was recalcitrant to biodegradation. In the medium containing 1 mM malic acid and 0.05-0.5 mM Eu(III), degradation of malic acid was not observed. The effect of metals on degradation of malic acid was in the order of Fe(III) < Al(III) < Eu(III). The stability constants of 1:1 Fe(III)-, Al(III)-, and Eu(III)-malic acid complexes are 7.1, 4.6, and 4.9, respectively. These results indicate that degradability of malic acid does not depend on the stability constants of metal-malic acid complexes. We found that 10 mM malic acid was degraded in the presence of 0.05 and 0.1 mM Eu(III) but 1 mM malic acid was not degraded in the presence of 0.05 and 0.1 mM Eu(III). The degradation rate of malic acid increased with a decreasing ratio of Eu(III) to malic acid. (authors)

  10. Pseudomonas fluorescens HK44: Lessons Learned from a Model Whole-Cell Bioreporter with a Broad Application History

    Directory of Open Access Journals (Sweden)

    Gary S. Sayler

    2012-02-01

    Full Text Available Initially described in 1990, Pseudomonas fluorescens HK44 served as the first whole-cell bioreporter genetically endowed with a bioluminescent (luxCDABE phenotype directly linked to a catabolic (naphthalene degradative pathway. HK44 was the first genetically engineered microorganism to be released in the field to monitor bioremediation potential. Subsequent to that release, strain HK44 had been introduced into other solids (soils, sands, liquid (water, wastewater, and volatile environments. In these matrices, it has functioned as one of the best characterized chemically-responsive environmental bioreporters and as a model organism for understanding bacterial colonization and transport, cell immobilization strategies, and the kinetics of cellular bioluminescent emission. This review summarizes the characteristics of P. fluorescens HK44 and the extensive range of its applications with special focus on the monitoring of bioremediation processes and biosensing of environmental pollution.

  11. The effect of rhamnolipid biosurfactant produced by Pseudomonas fluorescens on model bacterial strains and isolates from industrial wastewater.

    Science.gov (United States)

    Vasileva-Tonkova, Evgenia; Sotirova, Anna; Galabova, Danka

    2011-02-01

    In this study, the effect of rhamnolipid biosurfactant produced by Pseudomonas fluorescens on bacterial strains, laboratory strains, and isolates from industrial wastewater was investigated. It was shown that biosurfactant, depending on the concentration, has a neutral or detrimental effect on the growth and protein release of model Gram (+) strain Bacillus subtilis 168. The growth and protein release of model Gram (-) strain Pseudomonas aeruginosa 1390 was not influenced by the presence of biosurfactant in the medium. Rhamnolipid biosurfactant at the used concentrations supported the growth of some slow growing on hexadecane bacterial isolates, members of the microbial community. Changes in cell surface hydrophobicity and permeability of some Gram (+) and Gram (-) isolates in the presence of rhamnolipid biosurfactant were followed in experiments in vitro. It was found that bacterial cells treated with biosurfactant became more or less hydrophobic than untreated cells depending on individual characteristics and abilities of the strains. For all treated strains, an increase in the amount of released protein was observed with increasing the amount of biosurfactant, probably due to increased cell permeability as a result of changes in the organization of cell surface structures. The results obtained could contribute to clarify the relationships between members of the microbial community as well as suggest the efficiency of surface properties of rhamnolipid biosurfactant from Pseudomonas fluorescens making it potentially applicable in bioremediation of hydrocarbon-polluted environments.

  12. No apparent costs for facultative antibiotic production by the soil bacterium Pseudomonas fluorescens Pf0-1.

    Science.gov (United States)

    Garbeva, Paolina; Tyc, Olaf; Remus-Emsermann, Mitja N P; van der Wal, Annemieke; Vos, Michiel; Silby, Mark; de Boer, Wietse

    2011-01-01

    Many soil-inhabiting bacteria are known to produce secondary metabolites that can suppress microorganisms competing for the same resources. The production of antimicrobial compounds is expected to incur fitness costs for the producing bacteria. Such costs form the basis for models on the co-existence of antibiotic-producing and non-antibiotic producing strains. However, so far studies quantifying the costs of antibiotic production by bacteria are scarce. The current study reports on possible costs, for antibiotic production by Pseudomonas fluorescens Pf0-1, a soil bacterium that is induced to produce a broad-spectrum antibiotic when it is confronted with non-related bacterial competitors or supernatants of their cultures. We measured the possible cost of antibiotic production for Pseudomonas fluorescens Pf0-1 by monitoring changes in growth rate with and without induction of antibiotic production by supernatant of a bacterial competitor, namely Pedobacter sp.. Experiments were performed in liquid as well as on semi-solid media under nutrient-limited conditions that are expected to most clearly reveal fitness costs. Our results did not reveal any significant costs for production of antibiotics by Pseudomonas fluorescens Pf0-1. Comparison of growth rates of the antibiotic-producing wild-type cells with those of non-antibiotic producing mutants did not reveal costs of antibiotic production either. Based on our findings we propose that the facultative production of antibiotics might not be selected to mitigate metabolic costs, but instead might be advantageous because it limits the risk of competitors evolving resistance, or even the risk of competitors feeding on the compounds produced.

  13. No apparent costs for facultative antibiotic production by the soil bacterium Pseudomonas fluorescens Pf0-1.

    Directory of Open Access Journals (Sweden)

    Paolina Garbeva

    Full Text Available BACKGROUND: Many soil-inhabiting bacteria are known to produce secondary metabolites that can suppress microorganisms competing for the same resources. The production of antimicrobial compounds is expected to incur fitness costs for the producing bacteria. Such costs form the basis for models on the co-existence of antibiotic-producing and non-antibiotic producing strains. However, so far studies quantifying the costs of antibiotic production by bacteria are scarce. The current study reports on possible costs, for antibiotic production by Pseudomonas fluorescens Pf0-1, a soil bacterium that is induced to produce a broad-spectrum antibiotic when it is confronted with non-related bacterial competitors or supernatants of their cultures. METHODOLOGY AND PRINCIPAL FINDINGS: We measured the possible cost of antibiotic production for Pseudomonas fluorescens Pf0-1 by monitoring changes in growth rate with and without induction of antibiotic production by supernatant of a bacterial competitor, namely Pedobacter sp.. Experiments were performed in liquid as well as on semi-solid media under nutrient-limited conditions that are expected to most clearly reveal fitness costs. Our results did not reveal any significant costs for production of antibiotics by Pseudomonas fluorescens Pf0-1. Comparison of growth rates of the antibiotic-producing wild-type cells with those of non-antibiotic producing mutants did not reveal costs of antibiotic production either. SIGNIFICANCE: Based on our findings we propose that the facultative production of antibiotics might not be selected to mitigate metabolic costs, but instead might be advantageous because it limits the risk of competitors evolving resistance, or even the risk of competitors feeding on the compounds produced.

  14. IMPACT OF SIPHONING ACTIVITY AND NATURALLY SUSPENDED PARTICLE LOAD ON MUSSEL KILL by PSEUDOMONAS FLUORESCENS

    International Nuclear Information System (INIS)

    Daniel Molloy

    2003-01-01

    Under this USDOE-NETL contract, the bacterium Pseudomonas fluorescens is being developed as a biocontrol agent for zebra mussels. The specific purpose of the contract is to identify biotic and abiotic factors that affect mussel kill. Ingestion of these bacteria by zebra mussels is required to achieve kill, and tests evaluating factors that relate to mussel feeding are contained in this report. Specifically the impact of the following two factors were investigated: (1) Mussel siphoning behavior--In nature, zebra mussels typically have their two shells spread apart and their inhalant siphon tube extended from between their shells for taking food particles into their mantle cavities (Fig. 1). Our tests indicated that there is a direct correlation between mussel siphoning activity and mussel mortality achieved by a bacterial treatment. Therefore, to encourage mussel feeding on bacteria, future pipe treatments within power plants should be carried out using procedures which minimize disturbance to mussel siphoning. 2. Naturally suspended particle loads--Since bacterial cells are lethal only if ingested by mussels, waters containing very high levels of naturally suspended particles might reduce the mortality that can be achieved by a bacterial treatment. If true, this inhibition might occur as a result of particle exclusion, i.e., there could be reduced ingestion of bacterial cells since they represent a reduced percentage of all particles ingested. Our tests indicated that a range of particle concentrations that might naturally exist in a turbid river did not inhibit mussel kill by the bacterial cells, but that an artificially high load of natural particles was capable of causing a reduction in kill. To be conservative, therefore, future pipe treatments should be timed to occur when intake waters have relatively low quantities of naturally suspended particulate matter

  15. Biosurfactant production by Pseudomonas fluorescens growing on molasses and its application in phenol degradation

    Science.gov (United States)

    Suryantia, Venty; Marliyana, Soerya Dewi; Wulandari, Astri

    2015-12-01

    A molasses based medium for the biosurfactant production by Pseudomonas fluorescens was developed, where the effect of pre-treated of molasses and medium composition were evaluated. Biosurfactant production was followed by measuring optical density (OD), surface tension and emulsifying index (E24) over 12 days of fermentation. The optimum condition for the biosurfactant production was obtained when a medium containing of 8 g/L nutrient broth, 5 g/L NaCl, 1 g/L NH4NO3 and 5% v/v pre-treated molasses with centrifugation was used as media with 3 days of fermentation. The biosurfactant was identified as a rhamnolipid type biosurfactant which had critical micelle concentration (CMC) value of 801 mg/L and was able to reduce the surface tension of the water from 80 mN/m to 51 mN/m. The biosurfactants had water in oil (w/o) emulsion type. Biosurfactant was able to emulsify various hydrocarbons, which were able to decrase the interfacial tension about 50-75% when benzyl chloride, anisaldehyde and palm oil were used as immiscible compounds. The biosurfactant exhibited the E24 value of about 50% and the stable emulsion was reached up to 30 days when lubricant was used as an immiscible compound. Up to 68% of phenol was degraded in the presence of biosurfactant within 15 days, whereas only 56% of phenol was degraded in the absence of biosurfactant. Overall, the results exhibited that molasses are recommended for the rhamnolipids production which possessed good surface-active properties and had potential application in the enhancement of phenol degradation.

  16. D-glucose-6-phosphate dehydrogenase (Entner-Doudoroff enzyme) from Pseudomonas fluorescens

    International Nuclear Information System (INIS)

    Lessmann, D.; Schimz, K.L.; Kurz, G.

    1975-01-01

    The existence of two different D-glucose-6-phosphate dehydrogenases in Pseudomonas fluorescens has been demonstrated. Based on their different specificity and their different metabolic regulation one enzyme is appointed to the Entner-Doudoroff pathway and the other to the hexose monophosphate pathway. A procedure is described for the isolation of that D-glucose-6-phosphate dehydrogenase which forms part of the Entner-Doudoroff pathway (Entner-Doudoroff enzyme). A 950-fold purification was achieved with an overall yield of 44%. The final preparation, having a specific activity of about 300μmol NADH formed per min per mg protein, was shown to be homogeneous. The molecular weight of the Entner-Doudoroff enzyme has been determined to be 220,000 by gel permeation chromatography, and that of the other enzyme (Zwischenferment) has been shown to be 265,000. The pI of the Entner-Doudoroff enzyme has been shown to be 5.24 and that of the Zwischenferment 4.27. The Entner-Doudoroff enzyme is stable in the range of pH 6 to 10.5 and shows its maximal acivity at pH 8.9. The Entner-Doudoroff enzyme showed specificity for NAD + as well as for NADP + and exhibited homotropic effects for D-glucose 6-phosphate. It is inhibited by ATP which acts as a negative allosteric effector. Other nucleoside triphosphates as well as ADP are also inhibitory. The enzyme catalyzes the transfer of the axial hydrogen at carbon-1 of β-D-glucopyranose 6-phosphate to the si face of carbon-4 of the nicotinamide ring and must be classified as B-side stereospecific dehydrogenase. (orig.) [de

  17. Biodegradation of phenol, salicylic acid, benzenesulfonic acid, and iomeprol by Pseudomonas fluorescens in the capillary fringe.

    Science.gov (United States)

    Hack, Norman; Reinwand, Christian; Abbt-Braun, Gudrun; Horn, Harald; Frimmel, Fritz H

    2015-12-01

    Mass transfer and biological transformation phenomena in the capillary fringe were studied using phenol, salicylic acid, benzenesulfonic acid, and the iodinated X-ray contrast agent iomeprol as model organic compounds and the microorganism strain Pseudomonas fluorescens. Three experimental approaches were used: Batch experiments (uniform water saturation and transport by diffusion), in static columns (with a gradient of water saturation and advective transport in the capillaries) and in a flow-through cell (with a gradient of water saturation and transport by horizontal and vertical flow: 2-dimension flow-through microcosm). The reactors employed for the experiments were filled with quartz sand of defined particle size distribution (dp=200...600 μm, porosity ε=0.42). Batch experiments showed that phenol and salicylic acid have a high, whereas benzenesulfonic acid and iomeprol have a quite low potential for biodegradation under aerobic conditions and in a matrix nearly close to water saturation. Batch experiments under anoxic conditions with nitrate as electron acceptor revealed that the biodegradation of the model compounds was lower than under aerobic conditions. Nevertheless, the experiments showed that the moisture content was also responsible for an optimized transport in the liquid phase of a porous medium. Biodegradation in the capillary fringe was found to be influenced by both the moisture content and availability of the dissolved substrate, as seen in static column experiments. The gas-liquid mass transfer of oxygen also played an important role for the biological activity. In static column experiments under aerobic conditions, the highest biodegradation was found in the capillary fringe (e.g. βt/β0 (phenol)=0 after t=6 d) relative to the zone below the water table and unsaturated zone. The highest biodegradation occurred in the flow-through cell experiment where the height of the capillary fringe was largest. Copyright © 2015 Elsevier B.V. All rights

  18. Lipopeptide biosurfactant viscosin enhances dispersal of Pseudomonas fluorescens SBW25 biofilms.

    Science.gov (United States)

    Bonnichsen, Lise; Bygvraa Svenningsen, Nanna; Rybtke, Morten; de Bruijn, Irene; Raaijmakers, Jos M; Tolker-Nielsen, Tim; Nybroe, Ole

    2015-12-01

    Pseudomonads produce several lipopeptide biosurfactants that have antimicrobial properties but that also facilitate surface motility and influence biofilm formation. Detailed studies addressing the significance of lipopeptides for biofilm formation and architecture are rare. Hence, the present study sets out to determine the specific role of the lipopeptide viscosin in Pseudomonas fluorescens SBW25 biofilm formation, architecture and dispersal, and to relate viscA gene expression to viscosin production and effect. Initially, we compared biofilm formation of SBW25 and the viscosin-deficient mutant strain SBW25ΔviscA in static microtitre assays. These experiments demonstrated that viscosin had little influence on the amount of biofilm formed by SBW25 during the early stages of biofilm development. Later, however, SBW25 formed significantly less biofilm than SBW25ΔviscA. The indication that viscosin is involved in biofilm dispersal was confirmed by chemical complementation of the mutant biofilm. Furthermore, a fluorescent bioreporter showed that viscA expression was induced in biofilms 4 h prior to dispersal. Subsequent detailed studies of biofilms formed in flow cells for up to 5 days revealed that SBW25 and SBW25ΔviscA developed comparable biofilms dominated by well-defined, mushroom-shaped structures. Carbon starvation was required to obtain biofilm dispersal in this system. Dispersal of SBW25 biofilms was significantly greater than of SBW25ΔviscA biofilms after 3 h and, importantly, carbon starvation strongly induced viscA expression, in particular for cells that were apparently leaving the biofilm. Thus, the present study points to a role for viscosin-facilitated motility in dispersal of SBW25 biofilms.

  19. IMPACT OF SIPHONING ACTIVITY AND NATURALLY SUSPENDED PARTICLE LOAD ON MUSSEL KILL by PSEUDOMONAS FLUORESCENS

    Energy Technology Data Exchange (ETDEWEB)

    Daniel Molloy

    2003-08-04

    Under this USDOE-NETL contract, the bacterium Pseudomonas fluorescens is being developed as a biocontrol agent for zebra mussels. The specific purpose of the contract is to identify biotic and abiotic factors that affect mussel kill. Ingestion of these bacteria by zebra mussels is required to achieve kill, and tests evaluating factors that relate to mussel feeding are contained in this report. Specifically the impact of the following two factors were investigated: (1) Mussel siphoning behavior--In nature, zebra mussels typically have their two shells spread apart and their inhalant siphon tube extended from between their shells for taking food particles into their mantle cavities (Fig. 1). Our tests indicated that there is a direct correlation between mussel siphoning activity and mussel mortality achieved by a bacterial treatment. Therefore, to encourage mussel feeding on bacteria, future pipe treatments within power plants should be carried out using procedures which minimize disturbance to mussel siphoning. 2. Naturally suspended particle loads--Since bacterial cells are lethal only if ingested by mussels, waters containing very high levels of naturally suspended particles might reduce the mortality that can be achieved by a bacterial treatment. If true, this inhibition might occur as a result of particle exclusion, i.e., there could be reduced ingestion of bacterial cells since they represent a reduced percentage of all particles ingested. Our tests indicated that a range of particle concentrations that might naturally exist in a turbid river did not inhibit mussel kill by the bacterial cells, but that an artificially high load of natural particles was capable of causing a reduction in kill. To be conservative, therefore, future pipe treatments should be timed to occur when intake waters have relatively low quantities of naturally suspended particulate matter.

  20. Extracellular Protease of Pseudomonas fluorescens CHA0, a Biocontrol Factor with Activity against the Root-Knot Nematode Meloidogyne incognita

    OpenAIRE

    Siddiqui, Imran Ali; Haas, Dieter; Heeb, Stephan

    2005-01-01

    In Pseudomonas fluorescens CHA0, mutation of the GacA-controlled aprA gene (encoding the major extracellular protease) or the gacA regulatory gene resulted in reduced biocontrol activity against the root-knot nematode Meloidogyne incognita during tomato and soybean infection. Culture supernatants of strain CHA0 inhibited egg hatching and induced mortality of M. incognita juveniles more strongly than did supernatants of aprA and gacA mutants, suggesting that AprA protease contributes to biocon...

  1. Extracellular Protease of Pseudomonas fluorescens CHA0, a Biocontrol Factor with Activity against the Root-Knot Nematode Meloidogyne incognita

    Science.gov (United States)

    Siddiqui, Imran Ali; Haas, Dieter; Heeb, Stephan

    2005-01-01

    In Pseudomonas fluorescens CHA0, mutation of the GacA-controlled aprA gene (encoding the major extracellular protease) or the gacA regulatory gene resulted in reduced biocontrol activity against the root-knot nematode Meloidogyne incognita during tomato and soybean infection. Culture supernatants of strain CHA0 inhibited egg hatching and induced mortality of M. incognita juveniles more strongly than did supernatants of aprA and gacA mutants, suggesting that AprA protease contributes to biocontrol. PMID:16151170

  2. A phosphate-starvation-inducible outermembrane protein of Pseudomonas fluorescens Ag1 as an immunological phosphate-starvation marker

    DEFF Research Database (Denmark)

    Leopold, Kristine; Jacobsen, Susanne; Nybroe, Ole

    1997-01-01

    A phosphate-starvation-inducible outer-membrane protein of Pseudomonas fluorescens Ag1, expressed at phosphate concentrations below0.08-0.13 mM, was purified and characterized. The purification method involved separation of outer-membrane proteins by SDS-PAGE andextraction of the protein from...... nitrocellulose or PVDF membranes after electrotransfer of proteins to the membranes. The N-terminal amino acidsequence of the purified protein, called Psi1, did not show homology to any known proteins, and in contrast to the phosphate-specific porin OprP ofP. aeruginosa its mobility in SDS-PAGE was not affected...

  3. Contribution of alginate and levan production to biofilm formation by Pseudomonas syringae

    DEFF Research Database (Denmark)

    Laue, H.; Schenk, A.; Li, H.

    2006-01-01

    formation, biofilms of Pseudomonas syringae strains with different EPS patterns were compared. The mucoid strain PG4180.muc, which produces levan and alginate, and its levan- and/or alginate-deficient derivatives all formed biofilms in the wells of microtitre plates and in flow chambers. Confocal laser...... by binding of the lectin from Naja mossambica to a fibrous structure in biofilms of all P. syringae derivatives. Production of the as yet uncharacterized additional EPS might be more important for biofilm formation than the syntheses of levan and alginate.......Exopolysaccharides (EPSs) play important roles in the attachment of bacterial cells to a surface and/or in building and maintaining the three-dimensional, complex structure of bacterial biofilms. To elucidate the spatial distribution and function of the EPSs levan and alginate during biofilm...

  4. A conformational landscape for alginate secretion across the outer membrane of Pseudomonas aeruginosa

    International Nuclear Information System (INIS)

    Tan, Jingquan; Rouse, Sarah L.; Li, Dianfan; Pye, Valerie E.; Vogeley, Lutz; Brinth, Alette R.; El Arnaout, Toufic; Whitney, John C.; Howell, P. Lynne; Sansom, Mark S. P.; Caffrey, Martin

    2014-01-01

    Crystal structures of the β-barrel porin AlgE reveal a mechanism whereby alginate is exported from P. aeruginosa for biofilm formation. The exopolysaccharide alginate is an important component of biofilms produced by Pseudomonas aeruginosa, a major pathogen that contributes to the demise of cystic fibrosis patients. Alginate exits the cell via the outer membrane porin AlgE. X-ray structures of several AlgE crystal forms are reported here. Whilst all share a common β-barrel constitution, they differ in the degree to which loops L2 and T8 are ordered. L2 and T8 have been identified as an extracellular gate (E-gate) and a periplasmic gate (P-gate), respectively, that reside on either side of an alginate-selectivity pore located midway through AlgE. Passage of alginate across the membrane is proposed to be regulated by the sequential opening and closing of the two gates. In one crystal form, the selectivity pore contains a bound citrate. Because citrate mimics the uronate monomers of alginate, its location is taken to highlight a route through AlgE taken by alginate as it crosses the pore. Docking and molecular-dynamics simulations support and extend the proposed transport mechanism. Specifically, the P-gate and E-gate are flexible and move between open and closed states. Citrate can leave the selectivity pore bidirectionally. Alginate docks stably in a linear conformation through the open pore. To translate across the pore, a force is required that presumably is provided by the alginate-synthesis machinery. Accessing the open pore is facilitated by complex formation between AlgE and the periplasmic protein AlgK. Alginate can thread through a continuous pore in the complex, suggesting that AlgK pre-orients newly synthesized exopolysaccharide for delivery to AlgE

  5. A conformational landscape for alginate secretion across the outer membrane of Pseudomonas aeruginosa

    Energy Technology Data Exchange (ETDEWEB)

    Tan, Jingquan [Trinity College, Dublin (Ireland); Rouse, Sarah L. [University of Oxford, South Parks Road, Oxford (United Kingdom); Li, Dianfan; Pye, Valerie E.; Vogeley, Lutz; Brinth, Alette R.; El Arnaout, Toufic [Trinity College, Dublin (Ireland); Whitney, John C.; Howell, P. Lynne [The Hospital for Sick Children, Toronto, Ontario (Canada); University of Toronto, Toronto, Ontario (Canada); Sansom, Mark S. P. [University of Oxford, South Parks Road, Oxford (United Kingdom); Caffrey, Martin, E-mail: martin.caffrey@tcd.ie [Trinity College, Dublin (Ireland)

    2014-08-01

    Crystal structures of the β-barrel porin AlgE reveal a mechanism whereby alginate is exported from P. aeruginosa for biofilm formation. The exopolysaccharide alginate is an important component of biofilms produced by Pseudomonas aeruginosa, a major pathogen that contributes to the demise of cystic fibrosis patients. Alginate exits the cell via the outer membrane porin AlgE. X-ray structures of several AlgE crystal forms are reported here. Whilst all share a common β-barrel constitution, they differ in the degree to which loops L2 and T8 are ordered. L2 and T8 have been identified as an extracellular gate (E-gate) and a periplasmic gate (P-gate), respectively, that reside on either side of an alginate-selectivity pore located midway through AlgE. Passage of alginate across the membrane is proposed to be regulated by the sequential opening and closing of the two gates. In one crystal form, the selectivity pore contains a bound citrate. Because citrate mimics the uronate monomers of alginate, its location is taken to highlight a route through AlgE taken by alginate as it crosses the pore. Docking and molecular-dynamics simulations support and extend the proposed transport mechanism. Specifically, the P-gate and E-gate are flexible and move between open and closed states. Citrate can leave the selectivity pore bidirectionally. Alginate docks stably in a linear conformation through the open pore. To translate across the pore, a force is required that presumably is provided by the alginate-synthesis machinery. Accessing the open pore is facilitated by complex formation between AlgE and the periplasmic protein AlgK. Alginate can thread through a continuous pore in the complex, suggesting that AlgK pre-orients newly synthesized exopolysaccharide for delivery to AlgE.

  6. Comparative study on the in vitro effects of Pseudomonas aeruginosa and seaweed alginates on human gut microbiota.

    Directory of Open Access Journals (Sweden)

    Shaofeng Bai

    Full Text Available Alginates pertain to organic polysaccharides that have been extensively used in food- and medicine-related industries. The present study obtained alginates from an alginate overproducing Pseudomonas aeruginosa PAO1 mutant by screening transposon mutagenesis libraries. The interaction between bacterial and seaweed alginates and gut microbiota were further studied by using an in vitro batch fermentation system. Thin-layer chromatography (TLC analysis indicated that both bacterial and seaweed alginates can be completely degraded by fecal bacteria isolated from study volunteers, indicating that a minor structural difference between bacterial and seaweed alginates (O-acetylation and lack of G-G blocks didn't affect the digestion of alginates by human microbiota. Although, the digestion of bacterial and seaweed alginates was attributed to different Bacteroides xylanisolvens strains, they harbored similar alginate lyase genes. Genus Bacteroides with alginate-degrading capability were enriched in growth medium containing bacterial or seaweed alginates after in vitro fermentation. Short-chain fatty acid (SCFA production in both bacterial and seaweed alginates was also comparable, but was significantly higher than the same medium using starch. In summary, the present study has isolated an alginate-overproducing P. aeruginosa mutant strain. Both seaweed and bacterial alginates were degraded by human gut microbiota, and their regulatory function on gut microbiota was similar.

  7. Comparative study on the in vitro effects of Pseudomonas aeruginosa and seaweed alginates on human gut microbiota.

    Science.gov (United States)

    Bai, Shaofeng; Chen, Huahai; Zhu, Liying; Liu, Wei; Yu, Hongwei D; Wang, Xin; Yin, Yeshi

    2017-01-01

    Alginates pertain to organic polysaccharides that have been extensively used in food- and medicine-related industries. The present study obtained alginates from an alginate overproducing Pseudomonas aeruginosa PAO1 mutant by screening transposon mutagenesis libraries. The interaction between bacterial and seaweed alginates and gut microbiota were further studied by using an in vitro batch fermentation system. Thin-layer chromatography (TLC) analysis indicated that both bacterial and seaweed alginates can be completely degraded by fecal bacteria isolated from study volunteers, indicating that a minor structural difference between bacterial and seaweed alginates (O-acetylation and lack of G-G blocks) didn't affect the digestion of alginates by human microbiota. Although, the digestion of bacterial and seaweed alginates was attributed to different Bacteroides xylanisolvens strains, they harbored similar alginate lyase genes. Genus Bacteroides with alginate-degrading capability were enriched in growth medium containing bacterial or seaweed alginates after in vitro fermentation. Short-chain fatty acid (SCFA) production in both bacterial and seaweed alginates was also comparable, but was significantly higher than the same medium using starch. In summary, the present study has isolated an alginate-overproducing P. aeruginosa mutant strain. Both seaweed and bacterial alginates were degraded by human gut microbiota, and their regulatory function on gut microbiota was similar.

  8. Pseudomonas fluorescens filamentous hemagglutinin, an iron-regulated protein, is an important virulence factor that modulates bacterial pathogenicity

    Directory of Open Access Journals (Sweden)

    Yuan-yuan Sun

    2016-08-01

    Full Text Available Pseudomonas fluorescens is a common bacterial pathogen to a wide range of aquaculture animals including various species of fish. In this study, we employed proteomic analysis and identified filamentous hemagglutinin (FHA as an iron-responsive protein secreted by TSS, a pathogenic P. fluorescens isolate. In vitro study showed that compared to the wild type, the fha mutant TSSfha (i exhibited a largely similar vegetative growth profile but significantly retarded in the ability of biofilm growth and producing extracellular matrix, (ii displayed no apparent flagella and motility, (iii was defective in the attachment to host cells and unable to form self-aggregation, (iv displayed markedly reduced capacity of hemagglutination and surviving in host serum. In vivo infection analysis revealed that TSSfha was significantly attenuated in the ability of dissemination in fish tissues and inducing host mortality, and that antibody blocking of the natural FHA produced by the wild type TSS impaired the infectivity of the pathogen. Furthermore, when introduced into turbot as a subunit vaccine, recombinant FHA elicited a significant protection against lethal TSS challenge. Taken together, these results indicate for the first time that P. fluorescens FHA is a key virulence factor essential to multiple biological processes associated with pathogenicity.

  9. Listeria monocytogenes impact on mature or old Pseudomonas fluorescens biofilms during growth at 4 and 20ºC

    Directory of Open Access Journals (Sweden)

    Puga eCH

    2016-02-01

    Full Text Available Changes in spatial organization, as observed by confocal laser scanning microscopy (CLSM, viable cell content, biovolume and substratum surface coverage of the biofilms formed on glass by Pseudomonas fluorescens resulting from co-culture with Listeria monocytogenes, were examined. Two strains of L. monocytogenes, two culture temperatures and two biofilm developmental stages were investigated. Both L. monocytogenes strains, a persistently sampled isolate (collected repeatedly along 3 years from a meat factory and Scott A, induced shrinkage in matrix volume, both at 20ºC and 4ºC, in mature or old biofilms, without loss of P. fluorescens cell count per surface unit. The nearly homogeneous pattern of surface coverage shown by mono-species P. fluorescens biofilms, turned into more irregular layouts in co-culture with L. monocytogenes. The upper layer of both mono and dual-species biofilms turned to predominantly consist of matrix, with plenty of viable cells underneath, in old biofilms cultured at 20ºC, but not in those grown at 4ºC. Between 15 and 56% of the substratum area was covered by biofilm, the extent depending on temperature, time and L. monocytogenes strain. Real biofilms in food-related surfaces may thus be very heterogeneous regarding their superficial components, i.e. those more accessible to disinfectants. It is therefore a hygienic challenge to choose an adequate agent to disrupt them.

  10. Enlightening mineral iron sensing in Pseudomonas fluorescens by surface active maghemite nanoparticles: Involvement of the OprF porin.

    Science.gov (United States)

    Magro, Massimiliano; Fasolato, Luca; Bonaiuto, Emanuela; Andreani, Nadia Andrea; Baratella, Davide; Corraducci, Vittorino; Miotto, Giovanni; Cardazzo, Barbara; Vianello, Fabio

    2016-10-01

    Mineral iron(III) recognition by bacteria is considered a matter of debate. The peculiar surface chemistry of novel naked magnetic nanoparticles, called SAMNs (surface active maghemite nanoparticles) characterized by solvent exposed Fe(3+) sites on their surface, was exploited for studying mineral iron sensing in Pseudomonas fluorescens. SAMNs were applied for mimicking Fe(3+) ions in solution, acting as magnetically drivable probes to evaluate putative Fe(3+) recognition sites on the microorganism surface. Culture broths and nano-bio-conjugates were characterized by UV-Vis spectroscopy and mass spectrometry. The whole heritage of a membrane porin (OprF) of P. fluorescens Ps_22 cells was recognized and firmly bound by SAMNs. The binding of nanoparticles to OprF porin was correlated to a drastic inhibition of a siderophore (pyoverdine) biosynthesis and to the stimulation of the production and rate of formation of a secondary siderophore. The analysis of metabolic pathways, based on P. fluorescens Ps_22 genomic information, evidenced that this putative secondary siderophore does not belong to a selection of the most common siderophores. In the scenario of an adhesion mechanism, it is plausible to consider OprF as the biological component deputed to the mineral iron sensing in P. fluorescens Ps_22, as well as one key of siderophore regulation. The present work sheds light on mineral iron sensing in microorganisms. Peculiar colloidal naked iron oxide nanoparticles offer a useful approach for probing the adhesion of bacterial surface on mineral iron for the identification of the specific recognition site for this iron uptake regulation in microorganisms. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Effect of charged particle flux on Bacillus mesentericus and Pseudomonas fluorescens cultures

    International Nuclear Information System (INIS)

    Ostapenkov, A.M.; Kaptereva, Yu.V.; Merinov, N.S.; Lavrova, V.L.

    1979-01-01

    Studied was the effect of a strong electric field and charged particle (aerions) flow on Bac. mesentericus and Ps. fluorescens cultures causing the spoiling of foodstuffs and food raw materials. It has been found, that under the effects of the electric field and positive or negative ions reduced is the Bac. mesentericus and Ps. fluorescens viability, cultivated on a solid nutrient medium; the effect of the electric field only does not affect the Bac. mesentericus viability

  12. Alginate overproduction affects Pseudomonas aeruginosa biofilm structure and function

    DEFF Research Database (Denmark)

    Hentzer, Morten; Teitzel, G.M.; Balzer, G.J.

    2001-01-01

    -resistant communities of microorganisms organized in biofilms. Although biofilm formation and the conversion to mucoidy are both important aspects of CF pathogenesis, the relationship between them is at the present unclear. In this study, we report that the overproduction of alginate affects biofilm development...... on an abiotic surface. Biofilms formed by an alginate- overproducing strain exhibit a highly structured architecture and are significantly more resistant to the antibiotic tobramycin than a biofilm formed by an isogenic nonmucoid strain. These results suggest that an important consequence of the conversion...

  13. Type III secretion system and virulence markers highlight similarities and differences between human- and plant-associated Pseudomonads related to Pseudomonas fluorescens and P-putida

    OpenAIRE

    Mazurier, Sylvie; Merieau, Annabelle; Bergeau, Dorian; Decoin, Victorien; Sperandio, Daniel; Crepin, Alexandre; Barbey, Corinne; Jeannot, Katy; Vicre-Gibouin, Maité; Plesiat, Patrick; Lemanceau, Philippe; Latour, Xavier

    2015-01-01

    Pseudomonas fluorescens is commonly considered a saprophytic rhizobacterium devoid of pathogenic potential. Nevertheless, the recurrent isolation of strains from clinical human cases could indicate the emergence of novel strains originating from the rhizosphere reservoir, which could be particularly resistant to the immune system and clinical treatment. The importance of type three secretion systems (T3SSs) in the related Pseudomonas aeruginosa nosocomial species and the occurrence of this se...

  14. Draft Genome Sequences of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8, Soil Bacteria That Cooperate To Degrade the Poly- -D-Glutamic Acid Anthrax Capsule

    KAUST Repository

    Stabler, R. A.

    2013-01-24

    A mixed culture of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8 degraded poly-γ-d-glutamic acid; when the 2 strains were cultured separately, no hydrolytic activity was apparent. Here we report the draft genome sequences of both soil isolates.

  15. Draft Genome Sequences of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8, Soil Bacteria That Cooperate To Degrade the Poly-γ-d-Glutamic Acid Anthrax Capsule.

    Science.gov (United States)

    Stabler, Richard A; Negus, David; Pain, Arnab; Taylor, Peter W

    2013-01-01

    A mixed culture of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8 degraded poly-γ-d-glutamic acid; when the 2 strains were cultured separately, no hydrolytic activity was apparent. Here we report the draft genome sequences of both soil isolates.

  16. Draft Genome Sequences of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8, Soil Bacteria That Cooperate To Degrade the Poly-?-d-Glutamic Acid Anthrax Capsule

    OpenAIRE

    Stabler, Richard A.; Negus, David; Pain, Arnab; Taylor, Peter W.

    2013-01-01

    A mixed culture of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8 degraded poly-?-d-glutamic acid; when the 2 strains were cultured separately, no hydrolytic activity was apparent. Here we report the draft genome sequences of both soil isolates.

  17. Isolation and characterization of two new lipopeptide biosurfactants produced by Pseudomonas fluorescens BD5 isolated from water from the Arctic Archipelago of Svalbard

    Czech Academy of Sciences Publication Activity Database

    Janek, T.; Lukaszewicz, M.; Řezanka, Tomáš; Krasowska, A.

    2010-01-01

    Roč. 101, č. 15 (2010), s. 6118-6123 ISSN 0960-8524 Institutional research plan: CEZ:AV0Z50200510 Keywords : Pseudomonas fluorescens BD5 * Lipopeptide * Pseudofactins Subject RIV: EE - Microbiology, Virology Impact factor: 4.365, year: 2010

  18. Draft Genome Sequences of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8, Soil Bacteria That Cooperate To Degrade the Poly- -D-Glutamic Acid Anthrax Capsule

    KAUST Repository

    Stabler, R. A.; Negus, D.; Pain, Arnab; Taylor, P. W.

    2013-01-01

    A mixed culture of Pseudomonas fluorescens BS2 and Pusillimonas noertemannii BS8 degraded poly-γ-d-glutamic acid; when the 2 strains were cultured separately, no hydrolytic activity was apparent. Here we report the draft genome sequences of both soil isolates.

  19. Trichoderma harzianum enhances the production of nematicidal compounds in vitro and improves biocontrol of Meloidogyne javanica by Pseudomonas fluorescens in tomato.

    Science.gov (United States)

    Siddiqui, I A; Shaukat, S S

    2004-01-01

    To determine the influence of soil-borne fungus Trichoderma harzianum on the biocontrol performance of Pseudomonas fluorescens strain CHA0 and its 2,4-diacetylphloroglucinol (DAPG) overproducing derivative CHA0/pME3424 against Meloidogyne javanica. Amendment of the culture filtrate (CF) or methanol extract of the CF of a T. harzianum strain Th6 to P. fluorescens growth medium enhanced the production of nematicidal compound(s) by bacterial inoculants in vitro. In addition, bacteria overwhelmingly expressed phl'-'lacZ reporter gene when the medium was amended with CF of T. harzianum. Pseudomonas fluorescens and T. harzianum applied together in unsterilized sandy loam soil caused greater reduction in nematode population densities in tomato roots. Trichoderma harzianum improves root-knot nematode biocontrol by the antagonistic rhizobacterium P. fluorescens both in vitro and under glasshouse conditions. The synergistic effect of T. harzianum on the production of nematicidal compound(s) critical in biocontrol may improve the efficacy of biocontrol bacteria against plant-parasitic nematodes. Considering the inconsistent performance of the biocontrol agents under field conditions, application of a mixture of compatible T. harzianum and P. fluorescens would more closely mimic the natural situation and might broaden the spectrum of biocontrol activity with enhanced efficacy and reliability of control.

  20. Effect of Pseudomonas fluorescens and pyoverdine on the phytoextraction of cesium by red clover in soil pots and hydroponics.

    Science.gov (United States)

    Hazotte, Alice; Péron, Olivier; Gaudin, Pierre; Abdelouas, Abdesselam; Lebeau, Thierry

    2018-05-12

    With the aim of improving the phytoextraction rate of cesium (Cs), the effect of Pseudomonas fluorescens ATCC 17400 and its siderophore pyoverdine (PVD) on the uptake of Cs by red clover was studied in soil pots. This work also provides a mechanistic understanding of the Cs-bacteria (or PVD)-illite-plant interactions by using a simplified experimental design, i.e., hydroponics with either Cs in solution or Cs-spiked illite in suspension. For soil spiked with 11.2 mmol kg -1 (1480 mg kg -1 ) of Cs, 0.43% of total Cs was taken up by red clover in 12 days (119 μmol g -1 (16 mg g -1 ) of Cs dry matter in roots and 40 μmol g -1 (5 mg g -1 ) in shoots). In hydroponics with Cs in solution (0.1 mmol L -1 or 13 mg L -1 ), 75% of Cs was taken up vs. only 0.86% with Cs-spiked illite suspension. P. fluorescens and PVD did not increase Cs concentrations in aboveground parts and roots of red clover and even decreased them. The damaging effect of PVD on red clover growth was demonstrated with the biomass yielding 66% of the control in soil pots (and 100% mortality after 12 days of exposition) and only 56% in hydroponics (78% with illite in suspension). Nonetheless, PVD and, to a lesser extent, P. fluorescens increased the translocation factor up to a factor of 2.8. This study clearly showed a direct damaging effect of PVD and to a lower extent the retention of Cs by biofilm covering both the roots and illite, both resulting in the lower phytoextraction efficiency.

  1. Photocatalytic disinfection of spoilage bacteria Pseudomonas fluorescens and Macrococcus caseolyticus by nano-TiO2

    Science.gov (United States)

    Photocatalytic disinfection of spoilage bacteria gram-negative (G-) P. fluorescens and gram-positive (G+) M. caseolyticus by nano-TiO2 under different experimental conditions and the disinfection mechanism were investigated. The experimental conditions included the initial bacterial populations, nan...

  2. The study of Bacteriocin of Pseudomonas fluorescens and Citrus limon effects against Propionibacterium acnes and Staphylococcus epidermidis in acne patients

    Science.gov (United States)

    Ahmed, Mais E.

    2018-05-01

    Research was carried out on the antibacterial effect of (Citrus limon) juice on Acnevulgaris. Samples were obtained from individuals with pimples by swabbing their faces. Natural substances that derive from plants are promising to treat disease cause Acnevulgaris, the study in vitro biological activity of the juice, as well as bacterocin cultivated and fruits was investigated on two strains of bacteria (Propionibacterium acnes, Staphylococcus epidermidis). The new antimicrobial (bacteriocin and Citrus juice) is an ongoing search. This study used juice at different concentrations at (20%, 30%, 40%, 60%, 80% and 100%). The bacteriocin produced from local P. fluorescens isolates from wound infection and majority of isolates were found to produce crude bacteriocin were (P1 and P2) in Pseudomonas agar at 37°C for 24 hrs. Crude bacteriocin and Citrus limon juice against some pathogenic skin bacteria was find to be effective juice Citrus limon aganist S. epidermidis at 100% Concentrations with a range of inhibition zone (18) mm. The isolates of P. fluorescens (P2) was positive as producer of bacteriocin with a wide inhibition growth against gram positive pathogenic bacteria with a range between (10-12) mm.

  3. Mycorrhization between Cistus ladanifer L. and Boletus edulis Bull is enhanced by the mycorrhiza helper bacteria Pseudomonas fluorescens Migula.

    Science.gov (United States)

    Mediavilla, Olaya; Olaizola, Jaime; Santos-del-Blanco, Luis; Oria-de-Rueda, Juan Andrés; Martín-Pinto, Pablo

    2016-02-01

    Boletus edulis Bull. is one of the most economically and gastronomically valuable fungi worldwide. Sporocarp production normally occurs when symbiotically associated with a number of tree species in stands over 40 years old, but it has also been reported in 3-year-old Cistus ladanifer L. shrubs. Efforts toward the domestication of B. edulis have thus focused on successfully generating C. ladanifer seedlings associated with B. edulis under controlled conditions. Microorganisms have an important role mediating mycorrhizal symbiosis, such as some bacteria species which enhance mycorrhiza formation (mycorrhiza helper bacteria). Thus, in this study, we explored the effect that mycorrhiza helper bacteria have on the efficiency and intensity of the ectomycorrhizal symbiosis between C. ladanifer and B. edulis. The aim of this work was to optimize an in vitro protocol for the mycorrhizal synthesis of B. edulis with C. ladanifer by testing the effects of fungal culture time and coinoculation with the helper bacteria Pseudomonas fluorescens Migula. The results confirmed successful mycorrhizal synthesis between C. ladanifer and B. edulis. Coinoculation of B. edulis with P. fluorescens doubled within-plant mycorrhization levels although it did not result in an increased number of seedlings colonized with B. edulis mycorrhizae. B. edulis mycelium culture time also increased mycorrhization levels but not the presence of mycorrhizae. These findings bring us closer to controlled B. edulis sporocarp production in plantations.

  4. Novel ambler class A carbapenem-hydrolyzing beta-lactamase from a Pseudomonas fluorescens isolate from the Seine River, Paris, France.

    Science.gov (United States)

    Girlich, Delphine; Poirel, Laurent; Nordmann, Patrice

    2010-01-01

    A Pseudomonas fluorescens isolate (PF-1) resistant to carbapenems was recovered during an environmental survey performed with water from the Seine River (Paris). It expressed a novel Ambler class A carbapenemase, BIC-1, sharing 68 and 59% amino acid identities with beta-lactamases SFC-1 from Serratia fonticola and the plasmid-encoded KPC-2, respectively. beta-Lactamase BIC-1 hydrolyzed penicillins, carbapenems, and cephalosporins except ceftazidime and monobactams. The bla(BIC-1) gene was chromosomally located and was also identified in two other P. fluorescens strains isolated from the Seine River 3 months later.

  5. Exposure-related effects of Pseudomonas fluorescens, strain CL145A, on coldwater, coolwater, and warmwater fish

    Science.gov (United States)

    Luoma, James A.; Weber, Kerry L.; Denise A. Mayer,

    2015-01-01

    The exposure-related effects of a commercially prepared spray-dried powder (SDP) formulation of Pseudomonas fluorescens, strain CL145A, were evaluated on coldwater, coolwater, and warmwater fish endemic to the Great Lakes and Upper Mississippi River Basins. Nine species of young-of-the-year fish were exposed to SDP for 24 hours by using continuous-flow, serial-dilution exposure systems at temperatures of 12 degrees Celsius (°C; 2 species; Oncorhynchus mykiss [rainbow trout] and Salvelinus fontinalis [brook trout]), 17 °C (3 species; Perca flavescens [yellow perch], Sander vitreus [walleye], and Acipenser fulvescens [lake sturgeon]), or 22 °C (4 species; Micropterus salmoides [largemouth bass], Micropterus dolomieu [smallmouth bass], Lepomis macrochirus [bluegill sunfish], and Ictalurus punctatus [channel catfish]).

  6. One-step purification and characterization of alginate lyase from a clinical Pseudomonas aeruginosa with destructive activity on bacterial biofilm

    Directory of Open Access Journals (Sweden)

    Parinaz Ghadam

    2017-05-01

    Full Text Available Objective(s: Pseudomonas aeruginosais a Gram-negative and aerobic rod bacterium that displays mucoid and non-mucoid phenotype. Mucoid strains secrete alginate, which is the main agent of biofilms in chronic P. aeruginosa infections, show high resistance to antibiotics; consequently, the biological disruption of mucoid P. aeruginosa biofilms is an attractive area of study for researchers. Alginate lyase gene (algl is a member of alginate producing operon which by glycosidase activity produces primer for other enzymes in this cluster. Also this activity can destroy the extracellular alginate; therefore this enzyme participates in alginate production and destruction pathway. Alginate lyase causes detachment of a biofilm by reducing its adhesion to the surfaces, and increases phagocytosis and antibiotic susceptibility. In this study, alginate lyase was purified in just one step and its properties were investigated. Materials and Methods: The purification was done by affinity chromatography, analysed by SDS-PAGE, and its effect on P. aeruginosa biofilms was surveyed by micro titer plate assay and SEM. The substrate specificity of the enzyme was determined by PCR. Results: Alginate lyase from isolate 48 was purified in one step. It is more thermally resistant than alginate lyase from Pseudomonas aeruginosa PAO1 and poly M, poly G and poly MG alginate were the substrate of this enzyme. Moreover, it has an eradication effect on biofilms from P. aeruginosa 48 and PAO1. Conclusion: In this study an alginate lyase with many characteristics suitable in medicine such as thermal stability, effective on poly M alginate, and bacterial biofilm destructive was introduced and purified.

  7. Type III secretion system and virulence markers highlight similarities and differences between human- and plant-associated pseudomonads related to Pseudomonas fluorescens and P. putida.

    Science.gov (United States)

    Mazurier, Sylvie; Merieau, Annabelle; Bergeau, Dorian; Decoin, Victorien; Sperandio, Daniel; Crépin, Alexandre; Barbey, Corinne; Jeannot, Katy; Vicré-Gibouin, Maïté; Plésiat, Patrick; Lemanceau, Philippe; Latour, Xavier

    2015-04-01

    Pseudomonas fluorescens is commonly considered a saprophytic rhizobacterium devoid of pathogenic potential. Nevertheless, the recurrent isolation of strains from clinical human cases could indicate the emergence of novel strains originating from the rhizosphere reservoir, which could be particularly resistant to the immune system and clinical treatment. The importance of type three secretion systems (T3SSs) in the related Pseudomonas aeruginosa nosocomial species and the occurrence of this secretion system in plant-associated P. fluorescens raise the question of whether clinical isolates may also harbor T3SSs. In this study, isolates associated with clinical infections and identified in hospitals as belonging to P. fluorescens were compared with fluorescent pseudomonads harboring T3SSs isolated from plants. Bacterial isolates were tested for (i) their genetic relationships based on their 16S rRNA phylogeny, (ii) the presence of T3SS genes by PCR, and (iii) their infectious potential on animals and plants under environmental or physiological temperature conditions. Two groups of bacteria were delineated among the clinical isolates. The first group encompassed thermotolerant (41°C) isolates from patients suffering from blood infections; these isolates were finally found to not belong to P. fluorescens but were closely related and harbored highly conserved T3SS genes belonging to the Ysc-T3SS family, like the T3SSs from P. aeruginosa. The second group encompassed isolates from patients suffering from cystic fibrosis; these isolates belonged to P. fluorescens and harbored T3SS genes belonging to the Hrp1-T3SS family found commonly in plant-associated P. fluorescens. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  8. Genetic responses induced in olive roots upon colonization by the biocontrol endophytic bacterium Pseudomonas fluorescens PICF7.

    Directory of Open Access Journals (Sweden)

    Elisabetta Schilirò

    Full Text Available Knowledge on the genetic basis underlying interactions between beneficial bacteria and woody plants is still very limited, and totally absent in the case of olive. We aimed to elucidate genetic responses taking place during the colonization of olive roots by the native endophyte Pseudomonas fluorescens PICF7, an effective biocontrol agent against Verticillium wilt of olive. Roots of olive plants grown under non-gnotobiotic conditions were collected at different time points after PICF7 inoculation. A Suppression Subtractive Hybridization cDNA library enriched in induced genes was generated. Quantitative real time PCR (qRT-PCR analysis validated the induction of selected olive genes. Computational analysis of 445 olive ESTs showed that plant defence and response to different stresses represented nearly 45% of genes induced in PICF7-colonized olive roots. Moreover, quantitative real-time PCR (qRT-PCR analysis confirmed induction of lipoxygenase, phenylpropanoid, terpenoids and plant hormones biosynthesis transcripts. Different classes of transcription factors (i.e., bHLH, WRKYs, GRAS1 were also induced. This work highlights for the first time the ability of an endophytic Pseudomonas spp. strain to mount a wide array of defence responses in an economically-relevant woody crop such as olive, helping to explain its biocontrol activity.

  9. Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR‐type transcriptional regulator NunF

    OpenAIRE

    Hennessy, Rosanna C.; Phippen, Christopher B. W.; Nielsen, Kristian F.; Olsson, Stefan; Stougaard, Peter

    2017-01-01

    Abstract Nunamycin and nunapeptin are two antimicrobial cyclic lipopeptides (CLPs) produced by Pseudomonas fluorescens In5 and synthesized by nonribosomal synthetases (NRPS) located on two gene clusters designated the nun–nup regulon. Organization of the regulon is similar to clusters found in other CLP‐producing pseudomonads except for the border regions where putative LuxR‐type regulators are located. This study focuses on understanding the regulatory role of the LuxR‐type‐encoding gene nun...

  10. PENGARUH APLIKASI PSEUDOMONAS FLUORESCENS P60 TERHADAP MUTU PATOLOGIS, MUTU FISIOLOGIS, DAN PERTUMBUHAN BIBIT PADI IR 64

    Directory of Open Access Journals (Sweden)

    Lisa Navitasari

    2014-08-01

    Full Text Available Effect of Pseudomonas fluorescens P60 on pathological and physiological quality and growth of rice IR 64  seedlings. The research objectives were (1 detection and identification of seed-borne pathogens of IR 64 rice, (2 testing Pseudomonas fluorescents P60 in inhibiting the in vitro growth of seed-borne pathogens colonies, (3 testing P. fluorescents P60 for pathological and physiological seed quality, and (4 testing P. fluorescents P60 on the growth of seedlings in the greenhouse. The results showed that some seed-borne pathogens can be found both on farmers’ IR 64 rice and factory’s; they were Aspergillus flavus, Alternaria padwickii, Pseudomonas glumae, and P. syringae. Application of P. flourescens P60 was able to inhibit the in vitrogrowth of colonies of all seed-borne pathogens, except P. syringae.  Related to pathological quality, the effect of P. flourescens P60 on percentage of seed-borne pathogens attack did not significantly different from that of benomil but smaller than distilled water. On the physiological quality of seeds, treatment of P. flourescens P60 has the same effect with benomil and distilled water, with  germination rate was more than 80%. In the greenhouse study,treatment of seed immersion time  in P. flourescens P60 suspension showed that the effect of immersion time as long as15 minutes and 25 minutes on  seedling height, root length, and seedling dry weightdid not significantly different. were. However, 25 minutes immersion time resulted in fresh seedling weight and root dry weight higher than that of 15 minutes immersion time.

  11. The role of the antimicrobial compound 2,4-diacetylphloroglucinol in the impact of biocontrol Pseudomonas fluorescens F113 on Azospirillum brasilense phytostimulators.

    Science.gov (United States)

    Couillerot, Olivier; Combes-Meynet, Emeline; Pothier, Joël F; Bellvert, Floriant; Challita, Elita; Poirier, Marie-Andrée; Rohr, René; Comte, Gilles; Moënne-Loccoz, Yvan; Prigent-Combaret, Claire

    2011-06-01

    Pseudomonads producing the antimicrobial metabolite 2,4-diacetylphloroglucinol (Phl) can control soil-borne phytopathogens, but their impact on other plant-beneficial bacteria remains poorly documented. Here, the effects of synthetic Phl and Phl(+) Pseudomonas fluorescens F113 on Azospirillum brasilense phytostimulators were investigated. Most A. brasilense strains were moderately sensitive to Phl. In vitro, Phl induced accumulation of carotenoids and poly-β-hydroxybutyrate-like granules, cytoplasmic membrane damage and growth inhibition in A. brasilense Cd. Experiments with P. fluorescens F113 and a Phl(-) mutant indicated that Phl production ability contributed to in vitro growth inhibition of A. brasilense Cd and Sp245. Under gnotobiotic conditions, each of the three strains, P. fluorescens F113 and A. brasilense Cd and Sp245, stimulated wheat growth. Co-inoculation of A. brasilense Sp245 and Pseudomonas resulted in the same level of phytostimulation as in single inoculations, whereas it abolished phytostimulation when A. brasilense Cd was used. Pseudomonas Phl production ability resulted in lower Azospirillum cell numbers per root system (based on colony counts) and restricted microscale root colonization of neighbouring Azospirillum cells (based on confocal microscopy), regardless of the A. brasilense strain used. Therefore, this work establishes that Phl(+) pseudomonads have the potential to interfere with A. brasilense phytostimulators on roots and with their plant growth promotion capacity.

  12. The Effect of Bacteria Colony Pseudomonas fluorescens (UB_Pf1 and Bacillus subtilis (UB_Bs1 on the Mortality of Pratylenchus coffeae (Tylenchida: Pratylenchidae

    Directory of Open Access Journals (Sweden)

    Presti Mardiyani Purwaningtyas

    2016-11-01

    Full Text Available Parasitic Root-Lession nematode of Pratylenchus coffeae can reduce the Indonesian coffee plants productivity. Several studies reported that Pseudomonas fluorescens and Bacillus subtilis endophytic bacteria were antagonistic bacteria to nematode. The objective of this research was to reveal the effectiveness of bacterial colonies density of P. fluorescens (UB_Pf1, B.subtilis (UB BS1, and a combination of both bacteria on nematode mortality using median lethal concentration (LC50 and median lethal time 50 (LT50. The densities of bacteria used in this study were 107, 109, 1011 and 1013 cfu/ml. 35 testing nematodes were used and the mortality was counted at 6, 12, 24, 36, and 48 hours after treatments. The results showed that LC50 values of P. fluorescens was (UB_Pf1 was 4,3x108 cfu/ml, LC50 B. subtilis (UB_Bs1 was 1,9x109cfu/ ml, and LC50 combination of both bacteria was, 8x107 cfu/ml. It implies that the application of the combination of both bacteria are more pathogenic than single bacterial treatment. The results also showed that the highest LT50 value was 13.21  hours combination of bacterial colonies with a density of 1013 cfu/ml and the lowest LT50 value was 52.00 hours on P. fluorescens (UB_Pf1 treatment with colonies density of 107 cfu/ml.How to CitePurwaningtyas, P. M., Rahardjo, B. T., & Tarno, H. (2016. The Effect of Bacteria Colony Pseudomonas fluorescens (UB_Pf1 and Bacillus subtilis (UB_Bs1 on the Mortality of Pratylenchus coffeae (Tylenchida: Pratylenchidae. Biosaintifika: Journal of Biology & Biology Education, 8(3, 286-293. 

  13. Effect of Two Biological Formulations Based on Bacillus subtilis and Pseudomonas fluorescens on Control of Didymella applanata, the Causal Agent of Red Raspberry Cane Spur Blight

    Directory of Open Access Journals (Sweden)

    Margarita Shternshis

    2016-01-01

    Full Text Available In vitro and in vivo studies were conducted to estimate the efficacy of the two microbial formulations based on Bacillus subtilis Cohn. and Pseudomonas fluorescens Mig. on the fungus Didymella applanata (Niessl. Sacc., the causal agent of red raspberry (Rubus idaeus L. spur blight. In vitro, both bacteria reduced the growth of D. applanata. In inoculation experiments with raspberry canes in two cultivars with different susceptibility to D. applanata, these antagonistic bacteria suppressed fungal development by reducing the lesions area and the number of D. applanata fruiting bodies. Field trials of two biological formulations under natural conditions showed a significant suppression of the disease. B. subtilis and P. fluorescens included in the formulations revealed antagonistic activity towards D. applanata that depended on the red raspberry cultivar and weather conditions. In all cases, B. subtilis showed better results than P. fluorescens in biocontrol of the raspberry spur blight. This study demonstrated for the first time the ability of the biocontrol agents B. subtilis and P. fluorescens to suppress red raspberry cane spur blight, a serious worldwide disease.

  14. Conjoint effect of oil-seed cakes and Pseudomonas fluorescens on the growth of chickpea in relation to the management of plant-parasitic nematodes

    Directory of Open Access Journals (Sweden)

    Rose Rizvi

    2012-12-01

    Full Text Available Soil application of organics has been explored as an alternative means of organic management of plant-parasitic nematodes. Efficiency of different oil-seed cakes of neem (Azadirachta indica, castor (Ricinus communis, groundnut (Arachis hypogaea, linseed (Linum usitatissimum, sunflower (Helianthus annuus and soybean (Glycine max were evaluated in field conditions with association of Pseudomonas fluorescens in relation to growth parameters of chickpea and population of plant-parasitic nematodes. Their efficacious nature was highly effective in reducing the population of these dominant soil nematodes. Significant improvement was observed in plant-growth parameters such as plant weight, percent pollen fertility, pod numbers, root-nodulation and chlorophyll content of chickpea, seemed to be due to reduction in disease incidence and might be due to growth promoting substances secreted by P. fluorescens. The multiplication rate of nematodes was less in the presence of P. fluorescens as compared to its absence. Most effective combination of P. fluorescens was observed with neem cake.

  15. Measurement of glucose utilization by Pseudomonas fluorescens that are free-living and that are attached to surfaces

    International Nuclear Information System (INIS)

    Fletcher, M.

    1986-01-01

    The assimilation and respiration of glucose by attached and free-living Pseudomonas fluorescens were compared. The attachment surfaces were polyvinylidene fluoride, polyethylene, and glass. Specific uptake of [ 1 C]glucose was determined after bacterial biomass was measured by (1) microscopic counts or (2) prelabelling of cells by providing [ 3 H]leucine as substrate, followed by dual-labelling scintillation counting. The glucose concentration was 1.4, 3.5, 5.5, 7.6, or 9.7 μM. Glucose assimilation by cells which became detached from the surfaces during incubation with glucose was also measured after the detached cells were collected by filtration. The composition of the substratum had no effect on the amount of glucose assimilated by attached cells. Glucose assimilation by attached cells exceeded that by free-living cells by a factor of between 2 and 5 or more, and respiration of glucose by surface-associated cells was greater than that by free-living bacteria. Glucose assimilation by detached cells was greater than that by attached bacteria. Measurements of biomass by microscopic counts gave more consistent results than those obtained with dual-labelling, but in general, results obtained by both methods were corroborative

  16. Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR-type transcriptional regulator NunF

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Phippen, Christopher; Nielsen, Kristian Fog

    2017-01-01

    -producing pseudomonads except for the border regions where putative LuxR-type regulators are located. This study focuses on understanding the regulatory role of the LuxR-type-encoding gene nunF in CLP production of P. fluorescens In5. Functional analysis of nunF coupled with liquid chromatography-high-resolution mass......Nunamycin and nunapeptin are two antimicrobial cyclic lipopeptides (CLPs) produced by Pseudomonas fluorescens In5 and synthesized by nonribosomal synthetases (NRPS) located on two gene clusters designated the nun-nup regulon. Organization of the regulon is similar to clusters found in other CLP...... spectrometry (LC-HRMS) showed that CLP biosynthesis is regulated by nunF. Quantitative real-time PCR analysis indicated that transcription of the NRPS genes catalyzing CLP production is strongly reduced when nunF is mutated indicating that nunF is part of the nun-nup regulon. Swarming and biofilm formation...

  17. Structure of a putative BenF-like porin from Pseudomonas fluorescens Pf-5 at 2.6 A resolution

    Energy Technology Data Exchange (ETDEWEB)

    Sampathkumar, P.; Swaminathan, S.; Lu, F.; Zhao, X.; Li, Z.; Gilmore, J.; Bain, K.; Rutter, M. E.; Gheyi, T.; Schwinn, D.; Bonanno, J. B.; Pieper, U.; Fajardo, J. E.; Fiser, A.; Almo, S. C.; Chance, M. R.; Baker, D.; Atwell, S.; Thompson, D. A.; Emtage, J. S.; Wasserman, S. R.; Sali, A.; Sauder, J. M.; Burley, S. K.

    2010-11-01

    Gram-negative bacteria typically overcome poor permeability of outer membranes through general porins like OmpF and OmpC, which form water-filled transmembrane pores permitting diffusion of hydrophilic molecules with no particular selectivity. Many bacteria lacking such general porins use substrate-specific porins to overcome growth-limiting conditions and facilitate selective transport of metabolites. Exclusive reliance on substrate-specific porins yields lower membrane permeability to small molecules (<600 Da) versus that seen for Escherichia coli. In Pseudomonads, transit of most small molecules across the cell membrane is thought to be mediated by substrate-specific channels of the OprD superfamily. This property explains, at least in part, the high incidence of Pseudomonas aeruginosa antibiotic resistance. High-throughput DNA sequencing of the P. aeruginosa chromosome revealed the presence of 19 genes encoding structurally related, substrate-specific porins (with 30-45% pairwise amino acid sequence identity) that mediate transmembrane passage of small, water-soluble compounds. The OprD superfamily encompasses the eponymous OprD subfamily, which includes 9 P. aeruginosa proteins that convey basic amino acids and carbapenem antibiotics, and the OpdK subfamily, which includes 11 P. aeruginosa proteins that convey aromatic acids and other small aromatic compounds. Genome sequencing of other gram-negative bacteria has revealed additional members of the OprD and OpdK subfamilies in various organisms, including other pseudomonads. Among the many bacteria in which OprD superfamily members have been identified are P. putida, P. fluorescens Pf-5, P. syringae, and Azotobacter vinelandii, all of which share closely related genes that encode the so-called BenF-like porins. In P. putida, benF is part of an operon involved in benzoate catabolism regulated by benR. Within this operon, benK, benE, and benF genes have been suggested to contribute toward either influx or efflux

  18. Reduction of Fusarium wilt in watermelon by Pseudomonas chlororaphis PCL1391 and P. fluorescens WCS365

    Directory of Open Access Journals (Sweden)

    G.T. Tziros

    2007-12-01

    Full Text Available Fusarium wilt of watermelon (Citrullus lanatus caused by Fusarium oxysporum f. sp. niveum is a devastatine soil-borne disease that causes extensive losses throughout the world. Two known Pseudomonas biocontrol strains were used separately and in combination to assess their antagonistic effectiveness against F. oxysporum f. sp. niveum in pot experiments. P. chlororaphis PCL1391 signifi cantly reduced disease severity. P. fl uorescens WCS365 was less effective in disease suppression, while a combination of the two bacteria had intermediate effects. The biological control of Fusarium wilt with P. chlororaphis offers a potentially useful tool in an integrated pest management program to control Fusarium wilt of watermelon.

  19. Membrane-anchored MucR mediates nitrate-dependent regulation of alginate production in Pseudomonas aeruginosa

    KAUST Repository

    Wang, Yajie

    2015-04-29

    Alginates exhibit unique material properties suitable for medical and industrial applications. However, if produced by Pseudomonas aeruginosa, it is an important virulence factor in infection of cystic fibrosis patients. The alginate biosynthesis machinery is activated by c-di-GMP imparted by the inner membrane protein, MucR. Here, it was shown that MucR impairs alginate production in response to nitrate in P. aeruginosa. Subsequent site-specific mutagenesis of MucR revealed that the second MHYT sensor motif (MHYT II, amino acids 121–124) of MucR sensor domain was involved in nitrate sensing. We also showed that both c-di-GMP synthesizing and degrading active sites of MucR were important for alginate production. Although nitrate and deletion of MucR impaired alginate promoter activity and global c-di-GMP levels, alginate yields were not directly correlated with alginate promoter activity or c-di-GMP levels, suggesting that nitrate and MucR modulate alginate production at a post-translational level through a localized pool of c-di-GMP. Nitrate increased pel promoter activity in the mucR mutant while in the same mutant the psl promoter activity was independent of nitrate. Nitrate and deletion of mucR did not impact on swarming motility but impaired attachment to solid surfaces. Nitrate and deletion of mucR promoted the formation of biofilms with increased thickness, cell density, and survival. Overall, this study provided insight into the functional role of MucR with respect to nitrate-mediated regulation of alginate biosynthesis. © 2015 Springer-Verlag Berlin Heidelberg

  20. The biocontrol endophytic bacterium Pseudomonas fluorescens PICF7 induces systemic defense responses in aerial tissues upon colonization of olive roots

    Directory of Open Access Journals (Sweden)

    Carmen eGómez-Lama Cabanás

    2014-09-01

    Full Text Available Pseudomonas fluorescens PICF7, a native olive root endophyte and effective biocontrol agent (BCA against Verticillium wilt of olive, is able to trigger a broad range of defense responses in root tissues of this woody plant. In order to elucidate whether strain PICF7 also induces systemic defense responses in above-ground organs, aerial tissues of olive plants grown under non-gnotobiotic conditions were collected at different time points after root bacterization with this endophytic BCA. A suppression subtractive hybridization (SSH cDNA library, enriched in up-regulated genes, was generated. This strategy enabled the identification of 376 ESTs (99 contigs and 277 singlets, many of them related to response to different stresses. Five ESTs, involved in defense responses, were selected to carry out time-course quantitative real-time PCR (qRT-PCR experiments aiming to: (i validate the induction of these genes, and (ii shed light on their expression pattern along time (from 1 to 15 days. Induction of olive genes potentially coding for lypoxigenase 2, catalase, 1-aminocyclopropane-1-carboxylate oxidase and phenylananine ammonia-lyase was thus confirmed at some time points. Computational analysis also revealed that different transcription factors were up-regulated in olive aerial tissues (i.e. jerf, bHLH, WRKYs, as previously reported for roots. Results confirmed that root colonization by this endophytic bacterium does not only trigger defense responses in this organ but also mount a wide array of systemic defense responses in distant tissues (stems, leaves. This sheds light on how olive plants respond to the ‘non-hostile’ colonization by a bacterial endophyte and how induced defense response can contribute to the biocontrol activity of strain PICF7.

  1. Nunamycin and Nunapeptin: Two novel cyclic peptides are key components of the antimicrobial activity of the Greenlandic isolate Pseudomonas fluorescens In5

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Phippen, Christopher; Nielsen, Kristian F.

    Pseudomonas spp. are a rich source of secondary metabolites including bioactive non-ribosomal peptides (NRPs) and polyketides. NRPs are synthesised in large assembly lines by multi-domain modular enzymes known as NRP-synthetases (NRPS). Nunamycin and nunapeptin are two cyclic NRPs synthesised...... by the Greenlandic isolate P. fluorescens In5. Nunamycin shows antifungal activity against the basidiomycete Rhizoctonia solani whereas the only partially structure elucidated nunapeptin appears most active against the ascomycete Fusarium graminearum and the oomycete Pythium aphanidermatum. Originally isolated from...

  2. Pseudomonas fluorescens ATCC 13525 Containing an Artificial Oxalate Operon and Vitreoscilla Hemoglobin Secretes Oxalic Acid and Solubilizes Rock Phosphate in Acidic Alfisols

    OpenAIRE

    Yadav, Kavita; Kumar, Chanchal; Archana, G.; Naresh Kumar, G.

    2014-01-01

    Oxalate secretion was achieved in Pseudomonas fluorescens ATCC 13525 by incorporation of genes encoding Aspergillus niger oxaloacetate acetyl hydrolase (oah), Fomitopsis plaustris oxalate transporter (FpOAR) and Vitreoscilla hemoglobin (vgb) in various combinations. Pf (pKCN2) transformant containing oah alone accumulated 19 mM oxalic acid intracellularly but secreted 1.2 mM. However, in the presence of an artificial oxalate operon containing oah and FpOAR genes in plasmid pKCN4, Pf (pKCN4) s...

  3. Safety of spray-dried powder formulated Pseudomonas fluorescens strain CL145A exposure to subadult/adult unionid mussels during simulated open-water treatments

    Science.gov (United States)

    Luoma, James A.; Weber, Kerry L.; Waller, Diane L.; Wise, Jeremy K.; Mayer, Denise A.; Aloisi, Douglas B.

    2015-01-01

    The exposure effects of a commercially prepared spray dried powder (SDP) formulation ofPseudomonas fluorescens (strain CL145A) on the survival of seven species of unionid mussels endemic to the Great Lakes and Mississippi River basins was evaluated in this study. The study exposures were completed within replicated 350-liter test tanks contained within a mobile bioassay laboratory sited on the shores of the Black River near La Crosse, Wisconsin. The test tanks were supplied with flowing, filtered river water which was interrupted during the exposure period.

  4. Efeito da pré-incubação de Pseudomonas fluorescens em diferentes temperaturas de refrigeração sobre a hidrólise de caseína no leite desnatado pasteurizado microfiltrado

    OpenAIRE

    Netto, Gabriel Gama

    2012-01-01

    Este trabalho objetivou avaliar o efeito da multiplicação de uma estirpe psicrotrófica proteolítica de Pseudomonas fluorescens nas temperaturas de 4, 7 e 10 ºC, estabelecidas pela Instrução Normativa 62 do MAPA, nas características físicoquímicas e proteólise do leite microfiltrado. Amostras leite desnatado e pasteurizado foram inoculadas com a cepa Pseudomonas fluorescens 041 em uma concentração de aproximadamente 104 UFC mL-1 e incubadas nas três temperaturas por 48 h e, posteriormente, ...

  5. Investigating the ability of Pseudomonas fluorescens UW4 to reduce cadmium stress in Lactuca sativa via an intervention in the ethylene biosynthetic pathway.

    Science.gov (United States)

    Albano, Lucas J; Macfie, Sheila M

    2016-12-01

    A typical plant response to any biotic or abiotic stress, including cadmium (Cd), involves increased ethylene synthesis, which causes senescence of the affected plant part. Stressed plants can experience reduced ethylene and improved growth if they are inoculated with bacteria that have the enzyme ACC deaminase, which metabolizes the ethylene precursor ACC (1-aminocyclopropane-1-carboxylate). We investigated whether one such bacterium, Pseudomonas fluorescens UW4, reduces the production of ethylene and improves the growth of lettuce (Lactuca sativa) sown in Cd-contaminated potting material (PRO-MIX® BX). Plants were inoculated with the wild-type P. fluorescens UW4 or a mutant strain that cannot produce ACC deaminase. Cadmium-treated plants contained up to 50 times more Cd than did control plants. In noninoculated plants, Cd induced a 5-fold increase in ethylene concentration. The wild-type bacterium prevented Cd-induced reductions in root biomass but there was no relationship between Cd treatment and ethylene production in inoculated plants. In contrast, when the concentration of ethylene was plotted against the extent of bacterial colonization of the roots, increased colonization with wild-type P. fluorescens UW4 was associated with 20% less ethylene production. Ours is the first study to show that the protective effect of this bacterium is proportional to the quantity of bacteria on the root surface.

  6. Adsorption of Th(IV) and Pu(IV) on the surface of Pseudomonas fluorescens and Bacillus subtilis in the presence of desferrioxamine siderophore

    International Nuclear Information System (INIS)

    Yoshida, Takahiro; Ozaki, Takuo; Ohnuki, Toshihiko; Francis, Arokiasamy J.

    2005-01-01

    Adsorption of Th(IV) and Pu(IV) on a Gram-negative bacterium Pseudomonas fluorescens and a Gram-positive bacterium Bacillus subtilis in the presence of siderophore desferrioxamine B (DFO) was studied. Thorium(IV) and Pu(IV) were dissociated from DFO during adsorption on the cells. Thorium(IV) adsorption on bacterial cells in the presence of DFO was larger than that of Pu(IV) because of the smaller stability of the Th(IV)-DFO complex than that of the Pu(IV)-DFO complex. On the other hand, adsorption of Pu(IV) was larger than that of Fe(III), wherein the stability of the Pu(IV)- and Fe(III)-DFO complex is comparable. P. fluorescens showed a higher affinity for Th(IV) and Pu(IV) than B. subtilis, though potentiometric titration of bacterial cells indicated that surfaces of P. fluorescens and B. subtilis cells showed similar proton binding properties. (author)

  7. PRODUCTION AND CHARACTERIZATION OF BIOSURFACTANT BY Pseudomonas fluorescens USING CASSAVA FLOUR WASTEWATER AS MEDIA

    Directory of Open Access Journals (Sweden)

    Venty Suryanti

    2013-12-01

    Full Text Available Biosurfactant with efficient emulsification properties could be produced by Pseudomonas flourescens using cassava flour wastewater (manipueira as media. The ability of P. flourescens to produce biosurfactant could suggest potential use in industrial and environmental applications. Media containing a mixture of natural manipueira and nutrient broth with 48 h fermentation was the optimum condition for the biosurfactant production. Based on UV-Vis and FT-IR spectra, the biosurfactant was indicated as rhamnolipids containing hydroxyl, ester, carboxylic and aliphatic carbon chain functional groups. Biosurfactant exhibited critical micelle concentration (CMC value of 715 mg/L and reduced the surface tension of the water from 80 mN/m to 59 mN/m. The biosurfactant was able to decrease the interfacial tension about 51-70% when benzyl chloride, palm oil and kerosene were used as water-immiscible compounds. The biosurfactant was able to form stable emulsion until 30 days when paraffin, soybean oil, lubricant oil and kerosene were used as water-immiscible compounds.

  8. Non-pathogenic Fusarium solani represses the biosynthesis of nematicidal compounds in vitro and reduces the biocontrol of Meloidogyne javanica by Pseudomonas fluorescens in tomato.

    Science.gov (United States)

    Siddiqui, I A; Shaukat, S S

    2003-01-01

    The aim of the present investigation was to determine the influence of various Fusarium solani strains on the production of nematicidal agent(s) in vitro and biocontrol of Meloidogyne javanica in tomato by Pseudomonas fluorescens strains CHA0 and CHA0/pME3424. Culture filtrates (CF) of P. fluorescens strain CHA0 and its diacetylphloroglucinol-overproducing derivative CHA0/pME3424 caused substantial mortality of M. javanica juveniles in vitro. Bacterial growth medium amended with the growth medium of F. solani repressed the nematicidal activity of the bacteria. Methanol extract of F. solani CF resulting from Czapek's Dox liquid (CDL) medium without zinc amendment repressed the nematicidal activity of the bacteria while the CF obtained from CDL medium amended with zinc did not. Conidial suspension of F. solani strain Fs5 (repressor strain for the biosynthesis of nematicidal compounds in P. fluorescens) reduced biocontrol potential of the bacterial inoculants against M. javanica in tomato while strain Fs3 (non-repressor) did not. Fusarium solani strains with increased nematicidal activity repress the biosynthesis of nematicidal compounds by P. fluorescens strains in vitro and greatly alter its biocontrol efficacy against root-knot nematode under natural conditions. Fusarium solani strains are distributed worldwide and found in almost all the agricultural fields which suggest that some mycotoxin-producing strains will also be found in almost any soil sample taken. Besides the suppressive effect of these metabolite-producing strains on the production of nematicidal compound(s) critical in biocontrol, F. solani strains may also affect the performance of mycotoxin-sensitive biocontrol bacteria effective against plant-parasitic nematodes.

  9. 40 CFR 180.1107 - Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens; exemption from the requirement... killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens is exempt from the...

  10. Role of gallic and p-coumaric acids in the AHL-dependent expression of flgA gene and in the process of biofilm formation in food-associated Pseudomonas fluorescens KM120.

    Science.gov (United States)

    Myszka, Kamila; Schmidt, Marcin T; Białas, Wojciech; Olkowicz, Mariola; Leja, Katarzyna; Czaczyk, Katarzyna

    2016-09-01

    In the process of Pseudomonas fluorescens biofilm formation, N-acyl-l-homoserine lactone (AHL)-mediated flagella synthesis plays a key role. Inhibition of AHL production may attenuate P. fluorescens biofilm on solid surfaces. This work validated the anti-biofilm properties of p-coumaric and gallic acids via the ability of phenolics to suppress AHL synthesis in P. fluorescens KM120. The dependence between synthesis of AHL molecules, expression of flagella gene (flgA) and the ability of biofilm formation by P. fluorescens KM120 on a stainless steel surface (type 304L) was also investigated. Research was carried out in a purpose-built flow cell device. Limitations on AHL synthesis in P. fluorescens KM120 were observed at concentrations of 120 and 240 µmol L(-1) of phenolic acids in medium. At such levels of gallic and p-coumaric acids the ability of P. fluorescens KM120 to synthesize 3-oxo-C6-homoserine lactone (HSL) was not observed. These concentrations caused decreased expression of flgA gene in P. fluorescens KM120. The changes in expression of AHL-dependent flgA gene significantly decreased the rate of microorganism colonization on the stainless steel surface. Phenolic acids are able to inhibit biofilm formation. The results obtained in the work may help to develop alternative techniques for anti-biofilm treatment in the food industry. © 2015 Society of Chemical Industry. © 2015 Society of Chemical Industry.

  11. Interaction between the Bacterium Pseudomonas fluorescens strain CHA0, its genetic derivatives and vermiculite: Effects on chemical, mineralogical and mechanical properties of vermiculite

    Science.gov (United States)

    Mueller, Barbara

    2016-04-01

    Using bacteria of the strain Pseudomonas fluorescens wild type CHA0 and its genetic derivative strains CHA77, CHA89, CHA400, CHA631 and CHA661 (which differ in one gene only) the changes in chemical, mineralogical and rheological properties of the clay mineral vermiculite affected by microbial activity were studied in order to test whether the individually different production of metabolites by the genetically engineered strains may alter the clay mineral vermiculite in distinct ways. With the novel strategy of working with living wild type bacteria, their genetic derivatives and clay, the following properties of the mineral altered by the various strains of Pseudomonas fluorescens were determined: grain size, X-Ray diffraction pattern, intercrystalline swelling with glycerol, layer charge, CEC, BET surface and uptake of trace elements. Laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) was used to determine the changes in major, minor and trace elements of the clay vermiculite affected by microbial activity. Among all analyzed trace elements, Fe, Mn and Cu are the most interesting. Fe and Mn are taken up from the clay mineral by all bacterial strains whereas Cu is only removed from vermiculite by strains CHA0, CHA77, CHA400 and CHA661. The latter mentioned strains all produce the antibiotics 2,4-diacetylphloroglucinol and monoacetylphloroglucinol which can complex Cu efficiently. Therefore the alteration of only one gene of the bacteria is causing significant effects on the clay mineral.

  12. A full-length Plasmodium falciparum recombinant circumsporozoite protein expressed by Pseudomonas fluorescens platform as a malaria vaccine candidate.

    Directory of Open Access Journals (Sweden)

    Amy R Noe

    Full Text Available The circumsporozoite protein (CSP of Plasmodium falciparum is a major surface protein, which forms a dense coat on the sporozoite's surface. Preclinical research on CSP and clinical evaluation of a CSP fragment-based RTS, S/AS01 vaccine have demonstrated a modest degree of protection against P. falciparum, mediated in part by humoral immunity and in part by cell-mediated immunity. Given the partial protective efficacy of the RTS, S/AS01 vaccine in a recent Phase 3 trial, further improvement of CSP-based vaccines is crucial. In this report, we describe the preclinical development of a full-length, recombinant CSP (rCSP-based vaccine candidate against P. falciparum malaria suitable for current Good Manufacturing Practice (cGMP production. Utilizing a novel high-throughput Pseudomonas fluorescens expression platform, we demonstrated greater efficacy of full-length rCSP as compared to N-terminally truncated versions, rapidly down-selected a promising lead vaccine candidate, and developed a high-yield purification process to express immunologically active, intact antigen for clinical trial material production. The rCSP, when formulated with various adjuvants, induced antigen-specific antibody responses as measured by enzyme-linked immunosorbent assay (ELISA and immunofluorescence assay (IFA, as well as CD4+ T-cell responses as determined by ELISpot. The adjuvanted rCSP vaccine conferred protection in mice when challenged with transgenic P. berghei sporozoites containing the P. falciparum repeat region of CSP. Furthermore, heterologous prime/boost regimens with adjuvanted rCSP and an adenovirus type 35-vectored CSP (Ad35CS showed modest improvements in eliciting CSP-specific T-cell responses and anti-malarial protection, depending on the order of vaccine delivery. Collectively, these data support the importance of further clinical development of adjuvanted rCSP, either as a stand-alone product or as one of the components in a heterologous prime

  13. Role of ptsP, orfT, and sss recombinase genes in root colonization by Pseudomonas fluorescens Q8r1-96.

    Science.gov (United States)

    Mavrodi, Olga V; Mavrodi, Dmitri V; Weller, David M; Thomashow, Linda S

    2006-11-01

    Pseudomonas fluorescens Q8r1-96 produces 2,4-diacetylphloroglucinol (2,4-DAPG), a polyketide antibiotic that suppresses a wide variety of soilborne fungal pathogens, including Gaeumannomyces graminis var. tritici, which causes take-all disease of wheat. Strain Q8r1-96 is representative of the D-genotype of 2,4-DAPG producers, which are exceptional because of their ability to aggressively colonize and maintain large populations on the roots of host plants, including wheat, pea, and sugar beet. In this study, three genes, an sss recombinase gene, ptsP, and orfT, which are important in the interaction of Pseudomonas spp. with various hosts, were investigated to determine their contributions to the unusual colonization properties of strain Q8r1-96. The sss recombinase and ptsP genes influence global processes, including phenotypic plasticity and organic nitrogen utilization, respectively. The orfT gene contributes to the pathogenicity of Pseudomonas aeruginosa in plants and animals and is conserved among saprophytic rhizosphere pseudomonads, but its function is unknown. Clones containing these genes were identified in a Q8r1-96 genomic library, sequenced, and used to construct gene replacement mutants of Q8r1-96. Mutants were characterized to determine their 2,4-DAPG production, motility, fluorescence, colony morphology, exoprotease and hydrogen cyanide (HCN) production, carbon and nitrogen utilization, and ability to colonize the rhizosphere of wheat grown in natural soil. The ptsP mutant was impaired in wheat root colonization, whereas mutants with mutations in the sss recombinase gene and orfT were not. However, all three mutants were less competitive than wild-type P. fluorescens Q8r1-96 in the wheat rhizosphere when they were introduced into the soil by paired inoculation with the parental strain.

  14. 40 CFR 180.1108 - Delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens; exemption from the requirement... into killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens is...

  15. Pseudomonas fluorescens ATCC 13525 Containing an Artificial Oxalate Operon and Vitreoscilla Hemoglobin Secretes Oxalic Acid and Solubilizes Rock Phosphate in Acidic Alfisols

    Science.gov (United States)

    Archana, G.; Naresh Kumar, G.

    2014-01-01

    Oxalate secretion was achieved in Pseudomonas fluorescens ATCC 13525 by incorporation of genes encoding Aspergillus niger oxaloacetate acetyl hydrolase (oah), Fomitopsis plaustris oxalate transporter (FpOAR) and Vitreoscilla hemoglobin (vgb) in various combinations. Pf (pKCN2) transformant containing oah alone accumulated 19 mM oxalic acid intracellularly but secreted 1.2 mM. However, in the presence of an artificial oxalate operon containing oah and FpOAR genes in plasmid pKCN4, Pf (pKCN4) secreted 13.6 mM oxalate in the medium while 3.6 mM remained inside. This transformant solubilized 509 μM of phosphorus from rock phosphate in alfisol which is 4.5 fold higher than the Pf (pKCN2) transformant. Genomic integrants of P. fluorescens (Pf int1 and Pf int2) containing artificial oxalate operon (plac-FpOAR-oah) and artificial oxalate gene cluster (plac-FpOAR-oah, vgb, egfp) secreted 4.8 mM and 5.4 mM oxalic acid, released 329 μM and 351 μM P, respectively, in alfisol. The integrants showed enhanced root colonization, improved growth and increased P content of Vigna radiata plants. This study demonstrates oxalic acid secretion in P. fluorescens by incorporation of an artificial operon constituted of genes for oxalate synthesis and transport, which imparts mineral phosphate solubilizing ability to the organism leading to enhanced growth and P content of V. radiata in alfisol soil. PMID:24705024

  16. Pseudomonas fluorescens ATCC 13525 containing an artificial oxalate operon and Vitreoscilla hemoglobin secretes oxalic acid and solubilizes rock phosphate in acidic alfisols.

    Directory of Open Access Journals (Sweden)

    Kavita Yadav

    Full Text Available Oxalate secretion was achieved in Pseudomonas fluorescens ATCC 13525 by incorporation of genes encoding Aspergillus niger oxaloacetate acetyl hydrolase (oah, Fomitopsis plaustris oxalate transporter (FpOAR and Vitreoscilla hemoglobin (vgb in various combinations. Pf (pKCN2 transformant containing oah alone accumulated 19 mM oxalic acid intracellularly but secreted 1.2 mM. However, in the presence of an artificial oxalate operon containing oah and FpOAR genes in plasmid pKCN4, Pf (pKCN4 secreted 13.6 mM oxalate in the medium while 3.6 mM remained inside. This transformant solubilized 509 μM of phosphorus from rock phosphate in alfisol which is 4.5 fold higher than the Pf (pKCN2 transformant. Genomic integrants of P. fluorescens (Pf int1 and Pf int2 containing artificial oxalate operon (plac-FpOAR-oah and artificial oxalate gene cluster (plac-FpOAR-oah, vgb, egfp secreted 4.8 mM and 5.4 mM oxalic acid, released 329 μM and 351 μM P, respectively, in alfisol. The integrants showed enhanced root colonization, improved growth and increased P content of Vigna radiata plants. This study demonstrates oxalic acid secretion in P. fluorescens by incorporation of an artificial operon constituted of genes for oxalate synthesis and transport, which imparts mineral phosphate solubilizing ability to the organism leading to enhanced growth and P content of V. radiata in alfisol soil.

  17. Lipid transfer proteins and protease inhibitors as key factors in the priming of barley responses to Fusarium head blight disease by a biocontrol strain of Pseudomonas fluorescens.

    Science.gov (United States)

    Petti, Carloalberto; Khan, Mojibur; Doohan, Fiona

    2010-11-01

    Strains of non-pathogenic pseudomonad bacteria, can elicit host defence responses against pathogenic microorganisms. Pseudomonas fluorescens strain MKB158 can protect cereals from pathogenesis by Fusarium fungi, including Fusarium head blight which is an economically important disease due to its association with both yield loss and mycotoxin contamination of grain. Using the 22 K barley Affymetrix chip, trancriptome studies were undertaken to determine the local effect of P. fluorescens strain MKB158 on the transcriptome of barley head tissue, and to discriminate transcripts primed by the bacterium to respond to challenge by Fusarium culmorum, a causal agent of the economically important Fusarium head blight disease of cereals. The bacterium significantly affected the accumulation of 1203 transcripts and primed 74 to positively, and 14 to negatively, respond to the pathogen (P = 0.05). This is the first study to give insights into bacterium priming in the Triticeae tribe of grasses and associated transcripts were classified into 13 functional classes, associated with diverse functions, including detoxification, cell wall biosynthesis and the amplification of host defence responses. In silico analysis of Arabidopsis homologs of bacterium-primed barley genes indicated that, as is the case in dicots, jasmonic acid plays a role in pseudomonad priming of host responses. Additionally, the transcriptome studies described herein also reveal new insights into bacterium-mediated priming of host defences against necrotrophs, including the positive effects on grain filling, lignin deposition, oxidative stress responses, and the inhibition of protease inhibitors and proteins that play a key role in programmed cell death.

  18. Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR-type transcriptional regulator NunF.

    Science.gov (United States)

    Hennessy, Rosanna C; Phippen, Christopher B W; Nielsen, Kristian F; Olsson, Stefan; Stougaard, Peter

    2017-12-01

    Nunamycin and nunapeptin are two antimicrobial cyclic lipopeptides (CLPs) produced by Pseudomonas fluorescens In5 and synthesized by nonribosomal synthetases (NRPS) located on two gene clusters designated the nun-nup regulon. Organization of the regulon is similar to clusters found in other CLP-producing pseudomonads except for the border regions where putative LuxR-type regulators are located. This study focuses on understanding the regulatory role of the LuxR-type-encoding gene nunF in CLP production of P. fluorescens In5. Functional analysis of nunF coupled with liquid chromatography-high-resolution mass spectrometry (LC-HRMS) showed that CLP biosynthesis is regulated by nunF. Quantitative real-time PCR analysis indicated that transcription of the NRPS genes catalyzing CLP production is strongly reduced when nunF is mutated indicating that nunF is part of the nun-nup regulon. Swarming and biofilm formation was reduced in a nunF knockout mutant suggesting that these CLPs may also play a role in these phenomena as observed in other pseudomonads. Fusion of the nunF promoter region to mCherry showed that nunF is strongly upregulated in response to carbon sources indicating the presence of a fungus suggesting that environmental elicitors may also influence nunF expression which upon activation regulates nunamycin and nunapeptin production required for the growth inhibition of phytopathogens. © 2017 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.

  19. Methylobacterium sp. resides in unculturable state in potato tissues in vitro and becomes culturable after induction by Pseudomonas fluorescens IMGB163.

    Science.gov (United States)

    Podolich, O; Laschevskyy, V; Ovcharenko, L; Kozyrovska, N; Pirttilä, A M

    2009-03-01

    To induce growth of endophytic bacteria residing in an unculturable state in tissues of in vitro-grown potato plantlets. To isolate and identify the induced bacteria and to localize the strains in tissues of in vitro-grown potato plantlets. The inoculation of in vitro-grown potato plants with Pseudomonas fluorescens IMBG163 led to induction of another bacterium, a pink-pigmented facultative methylotroph that was identified as Methylobacterium sp. using phylogenetic 16S rDNA approach. Two molecular methods were used for localizing methylobacteria in potato plantlets: PCR and in situ hybridization (ISH/FISH). A PCR product specific for the Methylobacterium genus was found in DNA isolated from the surface-sterilized plantlet leaves. Presence of Methylobacterium rRNA was detected by ISH/FISH in leaves and stems of inoculated as well as axenic potato plantlets although the bacterium cannot be isolated from the axenic plants. Methylobacterium sp. resides in unculturable state within tissues of in vitro-grown potato plants and becomes culturable after inoculation with P. fluorescens IMBG163. In order to develop endophytic biofertilizers and biocontrol agents, a detailed knowledge of the life-style of endophytes is essential. To our knowledge, this is the first report on increase of the culturability of endophytes in response to inoculation by nonpathogenic bacteria.

  20. Characterisation of the thermostable protease AprX in strains of Pseudomonas fluorescens and impact on the shelf-life of dairy products: preliminary results

    Directory of Open Access Journals (Sweden)

    Nadia Andrea Andreani

    2016-12-01

    Full Text Available Bacterial proteases are involved in food spoilage and shelf-life reduction. Among the bacterial proteases, a predominant role in spoilage of dairy products seems to be played by the thermostable metallo-protease AprX, which is produced by various strains of Pseudomonas fluorescens. Differences in AprX enzyme activity among different strains were highlighted, but the most proteolytic strains were not identified. In this study, the presence of the aprX gene was evaluated in 69 strains isolated from food matrices and 18 reference strains belonging to the P. fluorescens group, which had been previously typed by the multi locus sequence typing method. Subsequently, a subset of reference strains was inoculated in ultra-high temperature milk, and the expression of the aprX gene was evaluated at 22 and 6°C. On the same milk samples, the proteolytic activity was then evaluated through Azocasein and trinitrobenzenesulfonic acid solution assays. Finally, to assess the applicability of the former assay directly on dairy products the proteolityc activity was tested on industrial ricotta samples using the Azocasein assay. These results demonstrate the spread of aprX gene in most strains tested and the applicability of Azocasein assay to monitor the proteolytic activity in dairy products.

  1. Non-target trials with Pseudomonas fluorescens strain CL145A, a lethal control agent of dreissenid mussels (Bivalvia: Dreissenidae

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    Daniel P. Molloy

    2013-01-01

    Full Text Available In an effort to develop an efficacious and environmentally safe method for managing zebra mussels (Dreissena polymorpha and quaggamussels (Dreissena rostriformis bugensis, we initiated a research project investigating the potential use of bacteria and their naturalmetabolic products as biocontrol agents. This project resulted in the discovery of an environmental isolate lethal to dreissenid mussels,Pseudomonas fluorescens strain CL145A (Pf-CL145A. In previous published reports we have demonstrated that: 1 Pf-CL145A’s mode ofaction is intoxication (not infection; 2 natural product within ingested bacterial cells lyse digestive tract epithelial cells leading to dreisseniddeath; and 3 high dreissenid kill rates (>90% are achievable following treatment with Pf-CL145A cells, irrespective of whether thebacterial cells are dead or alive. Investigating the environmental safety of Pf-CL145A was also a key element in our research efforts, andherein, we report the results of non-target trials demonstrating Pf-CL145A’s high specificity to dreissenids. These acute toxicity trials weretypically single-dose, short-term (24-72 h exposures to Pf-CL145A cells under aerated conditions at concentrations highly lethal todreissenids (100 or 200 mg/L. These trials produced no evidence of mortality among the ciliate Colpidium colpoda, the cladoceran Daphniamagna, three fish species (Pimephales promelas, Salmo trutta, and Lepomis macrochirus, and seven bivalve species (Mytilus edulis,Pyganodon grandis, Pyganodon cataracta, Lasmigona compressa, Strophitus undulatus, Lampsilis radiata, and Elliptio complanata. Lowmortality (3-27% was recorded in the amphipod Hyalella azteca, but additional trials suggested that most, if not all, of the mortality couldbe attributed to some other unidentified factor (e.g., possibly particle load or a water quality issue rather than Pf-CL145A’s dreissenidkillingnatural product. In terms of potential environmental safety, the results of

  2. Bacterial attack on phenolic ethers. Purification and characterization of the components of the meta O-dealkylase of Pseudomonas fluorescens Tp.

    Science.gov (United States)

    Pietrowski, R A; Cartwright, N J

    1977-01-01

    The meta O-dealkylase of Pseudomonas fluorescens Tp has been resolved into two protein components, neither of which is a cytochrome. The substrate binding terminal oxidase has been purified and shown to be a non-haem iron protein of approximate molecular weight 118,000, consisting of two seemingly identical subunits, each of molecular weight 55,000. Binding of substrate by the terminal oxidase has been established by difference spectroscopy. The amino acid composition of the protein has also been determined. The NADH-dependent reductase of the system has been partly purified and appears to have a molecular weight of 80,000. The similarity between this and other bacterial O-dealkylases is discussed.

  3. The periplasmic transaminase PtaA of Pseudomonas fluorescens converts the glutamic acid residue at the pyoverdine fluorophore to α-ketoglutaric acid.

    Science.gov (United States)

    Ringel, Michael T; Dräger, Gerald; Brüser, Thomas

    2017-11-10

    The periplasmic conversion of ferribactin to pyoverdine is essential for siderophore biogenesis in fluorescent pseudomonads, such as pathogenic Pseudomonas aeruginosa or plant growth-promoting Pseudomonas fluorescens The non-ribosomal peptide ferribactin undergoes cyclizations and oxidations that result in the fluorophore, and a strictly conserved fluorophore-bound glutamic acid residue is converted to a range of variants, including succinamide, succinic acid, and α-ketoglutaric acid residues. We recently discovered that the pyridoxal phosphate-containing enzyme PvdN is responsible for the generation of the succinamide, which can be hydrolyzed to succinic acid. Based on this, a distinct unknown enzyme was postulated to be responsible for the conversion of the glutamic acid to α-ketoglutaric acid. Here we report the identification and characterization of this enzyme in P. fluorescens strain A506. In silico analyses indicated a periplasmic transaminase in fluorescent pseudomonads and other proteobacteria that we termed PtaA for " p eriplasmic t ransaminase A " An in-frame-deleted ptaA mutant selectively lacked the α-ketoglutaric acid form of pyoverdine, and recombinant PtaA complemented this phenotype. The ptaA / pvdN double mutant produced exclusively the glutamic acid form of pyoverdine. PtaA is homodimeric and contains a pyridoxal phosphate cofactor. Mutation of the active-site lysine abolished PtaA activity and affected folding as well as Tat-dependent transport of the enzyme. In pseudomonads, the occurrence of ptaA correlates with the occurrence of α-ketoglutaric acid forms of pyoverdines. As this enzyme is not restricted to pyoverdine-producing bacteria, its catalysis of periplasmic transaminations is most likely a general tool for specific biosynthetic pathways. © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. Controlled ice nucleation using freeze-dried Pseudomonas syringae encapsulated in alginate beads.

    Science.gov (United States)

    Weng, Lindong; Tessier, Shannon N; Swei, Anisa; Stott, Shannon L; Toner, Mehmet

    2017-04-01

    The control of ice nucleation is of fundamental significance in many process technologies related to food and pharmaceutical science and cryobiology. Mechanical perturbation, electromagnetic fields and ice-nucleating agents (INAs) have been known to induce ice nucleation in a controlled manner. But these ice-nucleating methods may suffer from cumbersome manual operations, safety concerns of external fields, and biocompatibility and recovery issues of INA particles, especially when used in living systems. Given the automatic ice-seeding nature of INAs, a promising solution to overcome some of the above limitations is to engineer a biocomposite that accommodates the INA particles but minimizes their interactions with biologics, as well as enabling the recovery of used particles. In this study, freeze-dried Pseudomonas syringae, a model ice-nucleating agent, was encapsulated into microliter-sized alginate beads. We evaluated the performance of the bacterial hydrogel beads to initiate ice nucleation in water and aqueous glycerol solution by investigating factors including the size and number of the beads and the local concentration of INA particles. In the aqueous sample of a fixed volume, the total mass of the INA particles (m) was found to be the governing parameter that is solely responsible for determining the ice nucleation performance of the bacterial hydrogel beads. The freezing temperature has a strong positive linear correlation with log 10 m. The findings in this study provide an effective, predictable approach to control ice nucleation, which can improve the outcome and standardization of many ice-assisted process technologies. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Alginate overproduction and biofilm formation by psychrotolerant Pseudomonas mandelii depend on temperature in Antarctic marine sediments

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    Felipe Vásquez-Ponce

    2017-07-01

    Conclusion: Because biofilm formation is an efficient bacterial strategy to overcome stressful conditions, alginate overproduction might represent the best solution for the successful adaptation of P. mandelii to the extreme temperatures of the Antarctic. Through additional research, it is possible that this novel P. mandelii strain could become an additional source for biotechnological alginate production.

  6. Endophytic Bacterium Pseudomonas fluorescens RG11 May Transform Tryptophan to Melatonin and Promote Endogenous Melatonin Levels in the Roots of Four Grape Cultivars.

    Science.gov (United States)

    Ma, Yaner; Jiao, Jian; Fan, Xiucai; Sun, Haisheng; Zhang, Ying; Jiang, Jianfu; Liu, Chonghuai

    2016-01-01

    Endophytes have been verified to synthesize melatonin in vitro and promote abiotic stress-induced production of endogenous melatonin in grape ( Vitis vinifera L.) roots. This study aimed to further characterize the biotransformation of tryptophan to melatonin in the endophytic bacterium Pseudomonas fluorescens RG11 and to investigate its capacity for enhancing endogenous melatonin levels in the roots of different grape cultivars. Using ultra performance liquid chromatography-tandem mass spectrometry combined with 15N double-labeled L -tryptophan as the precursor for melatonin, we detected isotope-labeled 5-hydroxytryptophan, serotonin, N -acetylserotonin, and melatonin, but tryptamine was not detected during the in vitro incubation of P. fluorescens RG11. Furthermore, the production capacity of these four compounds peaked during the exponential growth phase. RG11 colonization increased the endogenous levels of 5-hydroxytryptophan, N -acetylserotonin, and melatonin, but reduced those of tryptamine and serotonin, in the roots of the Red Globe grape cultivar under salt stress conditions. Quantitative real-time PCR revealed that RG11 reduced the transcription of grapevine tryptophan decarboxylase and serotonin N -acetyltransferase genes when compared to the un-inoculated control. These results correlated with decreased reactive oxygen species bursts and cell damage, which were alleviated by RG11 colonization under salt stress conditions. Additionally, RG11 promoted plant growth and enhanced the levels of endogenous melatonin in different grape cultivars. Intraspecific variation in the levels of melatonin precursors was found among four grape cultivars, and the associated root crude extracts appeared to significantly induce RG11 melatonin biosynthesis in vitro . Overall, this study provides useful information that enhances the existing knowledge of a potential melatonin synthesis pathway in rhizobacteria, and it reveals plant-rhizobacterium interactions that affect

  7. Arabidopsis thaliana as a tool to identify traits involved in Verticillium dahliae biocontrol by the olive root endophyte Pseudomonas fluorescens PICF7

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    M. Mercedes eMaldonado-González

    2015-04-01

    Full Text Available The effective management of Verticillium wilts, diseases affecting many crops and caused by some species of the soil-borne fungus Verticillium, is problematic. The use of microbial antagonists to control these pathologies fits modern sustainable agriculture criteria. Pseudomonas fluorescens PICF7 is an endophytic bacterium isolated from olive roots with demonstrated ability to control Verticillium wilt of olive caused by the highly-virulent, defoliating (D pathotype of Verticillium dahliae Kleb. However, the study of the PICF7-V.dahliae-olive tripartite interaction poses difficulties because of the inherent characteristics of woody, long-living plants. To overcome these problems we explored the use of the model plant Arabidopsis thaliana. Results obtained in this study showed that: (i olive D and non-defoliating (ND V. dahliae pathotypes produce differential disease severity in A. thaliana plants; (ii strain PICF7 is able to colonize and persist in the A. thaliana rhizosphere but is not endophytic in Arabidopsis; and (iii strain PICF7 controls Verticillium wilt (VW in Arabidopsis. Additionally, as previously observed in olive, neither swimming motility nor siderophore production by PICF7 are required for VW control in A. thaliana, whilst cysteine auxotrophy decreased the effectiveness of PICF7. Moreover, when applied to the roots PICF7 controlled Botrytis cinerea infection in the leaves of Arabidopsis, suggesting that this strain is able to induce systemic resistance. Arabidopsis thaliana is therefore a suitable alternative to olive bioassays to unravel biocontrol traits involved in biological control of V. dahliae by P. fluorescens PICF7.

  8. Application of Endophytic Pseudomonas fluorescens and a Bacterial Consortium to Brassica napus Can Increase Plant Height and Biomass under Greenhouse and Field Conditions

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    Richard D. Lally

    2017-12-01

    Full Text Available Plant associated bacteria with plant growth promotion (PGP properties have been proposed for use as environmentally friendly biofertilizers for sustainable agriculture; however, analysis of their efficacy in the field is often limited. In this study, greenhouse and field trials were carried out using individual endophytic Pseudomonas fluorescens strains, the well characterized rhizospheric P. fluorescens F113 and an endophytic microbial consortium of 10 different strains. These bacteria had been previously characterized with respect to their PGP properties in vitro and had been shown to harbor a range of traits associated with PGP including siderophore production, 1-aminocyclopropane-1-carboxylic acid (ACC deaminase activity, and inorganic phosphate solubilization. In greenhouse experiments individual strains tagged with gfp and Kmr were applied to Brassica napus as a seed coat and were shown to effectively colonize the rhizosphere and root of B. napus and in addition they demonstrated a significant increase in plant biomass compared with the non-inoculated control. In the field experiment, the bacteria (individual and consortium were spray inoculated to winter oilseed rape B. napus var. Compass which was grown under standard North Western European agronomic conditions. Analysis of the data provides evidence that the application of the live bacterial biofertilizers can enhance aspects of crop development in B. napus at field scale. The field data demonstrated statistically significant increases in crop height, stem/leaf, and pod biomass, particularly, in the case of the consortium inoculated treatment. However, although seed and oil yield were increased in the field in response to inoculation, these data were not statistically significant under the experimental conditions tested. Future field trials will investigate the effectiveness of the inoculants under different agronomic conditions.

  9. Sequencing and Analysis of the Pseudomonas fluorescens GcM5-1A Genome: A Pathogen Living in the Surface Coat of Bursaphelenchus xylophilus.

    Directory of Open Access Journals (Sweden)

    Kai Feng

    Full Text Available It is known that several bacteria are adherent to the surface coat of pine wood nematode (Bursaphelenchus xylophilus, but their function and role in the pathogenesis of pine wilt disease remains debatable. The Pseudomonas fluorescens GcM5-1A is a bacterium isolated from the surface coat of pine wood nematodes. In previous studies, GcM5-1A was evident in connection with the pathogenicity of pine wilt disease. In this study, we report the de novo sequencing of the GcM5-1A genome. A 600-Mb collection of high-quality reads was obtained and assembled into sequence contigs spanning a 6.01-Mb length. Sequence annotation predicted 5,413 open reading frames, of which 2,988 were homologous to genes in the other four sequenced P. fluorescens isolates (SBW25, WH6, Pf0-1 and Pf-5 and 1,137 were unique to GcM5-1A. Phylogenetic studies and genome comparison revealed that GcM5-1A is more closely related to SBW25 and WH6 isolates than to Pf0-1 and Pf-5 isolates. Towards study of pathogenesis, we identified 79 candidate virulence factors in the genome of GcM5-1A, including the Alg, Fl, Waa gene families, and genes coding the major pathogenic protein fliC. In addition, genes for a complete T3SS system were identified in the genome of GcM5-1A. Such systems have proved to play a critical role in subverting and colonizing the host organisms of many gram-negative pathogenic bacteria. Although the functions of the candidate virulence factors need yet to be deciphered experimentally, the availability of this genome provides a basic platform to obtain informative clues to be addressed in future studies by the pine wilt disease research community.

  10. Colony morphology and transcriptome profiling of Pseudomonas putida KT2440 and its mutants deficient in alginate or all EPS synthesis under controlled matric potentials

    DEFF Research Database (Denmark)

    Gülez, Gamze; Altintas, Ali; Fazli, Mustafa

    2014-01-01

    Pseudomonas putida is a versatile bacterial species adapted to soil and its fluctuations. Like many other species living in soil, P. putida often faces water limitation. Alginate, an exopolysaccharide (EPS) produced by P. putida, is known to create hydrated environments and alleviate the effect o...

  11. Exploiting genotypic diversity of 2,4-diacetylphloroglucinol-producing Pseudomonas spp.: characterization of superior root-colonizing P. fluorescens strain Q8r1-96.

    Science.gov (United States)

    Raaijmakers, J M; Weller, D M

    2001-06-01

    The genotypic diversity that occurs in natural populations of antagonistic microorganisms provides an enormous resource for improving biological control of plant diseases. In this study, we determined the diversity of indigenous 2,4-diacetylphloroglucinol (DAPG)-producing Pseudomonas spp. occurring on roots of wheat grown in a soil naturally suppressive to take-all disease of wheat. Among 101 isolates, 16 different groups were identified by random amplified polymorphic DNA (RAPD) analysis. One RAPD group made up 50% of the total population of DAPG-producing Pseudomonas spp. Both short- and long-term studies indicated that this dominant genotype, exemplified by P. fluorescens Q8r1-96, is highly adapted to the wheat rhizosphere. Q8r1-96 requires a much lower dose (only 10 to 100 CFU seed(-1) or soil(-1)) to establish high rhizosphere population densities (10(7) CFU g of root(-1)) than Q2-87 and 1M1-96, two genotypically different, DAPG-producing P. fluorescens strains. Q8r1-96 maintained a rhizosphere population density of approximately 10(5) CFU g of root(-1) after eight successive growth cycles of wheat in three different, raw virgin soils, whereas populations of Q2-87 and 1M1-96 dropped relatively quickly after five cycles and were not detectable after seven cycles. In short-term studies, strains Q8r1-96, Q2-87, and 1M1-96 did not differ in their ability to suppress take-all. After eight successive growth cycles, however, Q8r1-96 still provided control of take-all to the same level as obtained in the take-all suppressive soil, whereas Q2-87 and 1M1-96 gave no control anymore. Biochemical analyses indicated that the superior rhizosphere competence of Q8r1-96 is not related to in situ DAPG production levels. We postulate that certain rhizobacterial genotypes have evolved a preference for colonization of specific crops. By exploiting diversity of antagonistic rhizobacteria that share a common trait, biological control can be improved significantly.

  12. Pseudomonas aeruginosa alginate is refractory to Th1 immune response and impedes host immune clearance in a mouse model of acute lung infection

    DEFF Research Database (Denmark)

    Song, Zhijun; Wu, Hong; Ciofu, Oana

    2003-01-01

    . The effect of alginate production on pathogenicity was investigated by using an acute lung infection mouse model that compared a non-mucoid P. aeruginosa strain, PAO1, to its constitutive alginate-overproducing derivative, Alg(+) PAOmucA22, and an alginate-defective strain, Alg(-) PAOalgD. Bacterial......Pseudomonas aeruginosa is an opportunistic respiratory pathogen that accounts for most of the morbidity and mortality in cystic fibrosis (CF) patients. In CF-affected lungs, the bacteria undergo conversion from a non-mucoid to a non-tractable mucoid phenotype, due to overproduction of alginate...... suspensions were instilled into the left bronchus and examined 24 and 48 h post-infection. The highest bacterial loads and the most severe lung pathology were observed with strain Alg(-) PAOalgD at 24 h post-infection, which may have been due to an increase in expression of bacterial elastase by the mutant...

  13. Impact of biocontrol Pseudomonas fluorescens CHA0 and a genetically modified derivative on the diversity of culturable fungi in the cucumber rhizosphere.

    Science.gov (United States)

    Girlanda, M; Perotto, S; Moenne-Loccoz, Y; Bergero, R; Lazzari, A; Defago, G; Bonfante, P; Luppi, A M

    2001-04-01

    Little is known about the effects of Pseudomonas biocontrol inoculants on nontarget rhizosphere fungi. This issue was addressed using the biocontrol agent Pseudomonas fluorescens CHA0-Rif, which produces the antimicrobial polyketides 2,4-diacetylphloroglucinol (Phl) and pyoluteorin (Plt) and protects cucumber from several fungal pathogens, including Pythium spp., as well as the genetically modified derivative CHA0-Rif(pME3424). Strain CHA0-Rif(pME3424) overproduces Phl and Plt and displays improved biocontrol efficacy compared with CHA0-Rif. Cucumber was grown repeatedly in the same soil, which was left uninoculated, was inoculated with CHA0-Rif or CHA0-Rif(pME3424), or was treated with the fungicide metalaxyl (Ridomil). Treatments were applied to soil at the start of each 32-day-long cucumber growth cycle, and their effects on the diversity of the rhizosphere populations of culturable fungi were assessed at the end of the first and fifth cycles. Over 11,000 colonies were studied and assigned to 105 fungal species (plus several sterile morphotypes). The most frequently isolated fungal species (mainly belonging to the genera Paecilomyces, Phialocephala, Fusarium, Gliocladium, Penicillium, Mortierella, Verticillium, Trichoderma, Staphylotrichum, Coniothyrium, Cylindrocarpon, Myrothecium, and Monocillium) were common in the four treatments, and no fungal species was totally suppressed or found exclusively following one particular treatment. However, in each of the two growth cycles studied, significant differences were found between treatments (e.g., between the control and the other treatments and/or between the two inoculation treatments) using discriminant analysis. Despite these differences in the composition and/or relative abundance of species in the fungal community, treatments had no effect on species diversity indices, and species abundance distributions fit the truncated lognormal function in most cases. In addition, the impact of treatments at the 32-day

  14. Polymorphism of a lipid extract from Pseudomonas fluorescens: Structure analysis of a hexagonal phase and of a novel cubic phase of extinction symbol Fd--

    International Nuclear Information System (INIS)

    Mariani, P.; Rivas, E.; Delacroix, H.; Luzzati, V.

    1990-01-01

    The phase diagram of the Pseudomonas fluorescens lipid extract is unusual, in the sense that it displays a cubic phase straddled by a hexagonal phase. The hexagonal phase was studied over an extended concentration range, and the reflections were phased on the assumption that the structure contains circular cylinders of known radius. The cubic phase, whose extinction symbol is Fd--, was analyzed by reference to space group No. 227 (Fd3m). The phases of the reflections were determined by using a novel pattern recognition approach, based upon the notion that the average fourth power of the electron density contrast 4 > is dependent on chemical composition but not on physical structure, provided that the function Δr(r) satisfies the constraints = 0 and 2 > = 1. The authors analyzed two cubic samples of different composition: for each of them they generated all the phase combinations compatible with the X-ray scattering data and they searched for those whose 4 > best agrees with the hexagonal phase. They concluded that the chemical composition of the phases being compared must be identical, that the X-ray scattering data should not be truncated artificially, and that the apodization must be mild so that the curvature takes a value intermediate between those corresponding to the raw data of the two phases. The structure may be visualized as a 3D generalization of the lipid monolayer. The structure, moreover, does not belong to the class of the infinite periodic surfaces without intersections

  15. Pseudomonas fluorescens JH 70-4 promotes pb stabilization and early seedling growth of sudan grass in contaminated mining site soil.

    Science.gov (United States)

    Shim, Jaehong; Babu, A Giridhar; Velmurugan, Palanivel; Shea, Patrick J; Oh, Byung-Taek

    2014-01-01

    A bacterial strain (JH 70-4) exhibiting plant growth promoting characteristics (indoleacetic acid production and 1-aminocyclopropane-1-carboxylate deaminase activity), as well as heavy metal(loid) (HM) tolerance and Pb precipitation, was isolated from HM-contaminated soil at an abandoned mine site. The bacterium was identified as Pseudomonas fluorescens based on 16S rDNA sequencing. The JH 70-4 strain induced precipitation of Pb as PbS nanoparticles, confirmed by X-ray diffraction. Solution pH, incubation time, and Pb concentration influenced removal and PbS formation. Inoculating contaminated soil with JH 70-4 decreased Pb availability; exchangeable Pb decreased while organic- and sulphide-bound Pb increased. The toxicity characteristic leaching procedure showed a 65% decrease in Pb in leachate 60 d after inoculating soil with JH 70-4. Shoot and root lengths of Sudan grass grown in the inoculated soil were greater than in the uninoculated soil. Findings suggest that microbial Pb fixation is a viable strategy for remediating soil and promoting plant growth for phytostabilization of contaminated sites.

  16. Synthesis of amino-silane modified superparamagnetic Fe{sub 3}O{sub 4} nanoparticles and its application in immobilization of lipase from Pseudomonas fluorescens Lp1

    Energy Technology Data Exchange (ETDEWEB)

    Kanimozhi, S., E-mail: skanimo@gmail.com [Department of Biotechnology, Sathyabama University, Jeppiaar Nagar, Rajivgandhi Salai, Chennai 600119, Tamil Nadu (India); Perinbam, K. [Department of Plant Biology and Biotechnology, Nandanam Arts College (Men), Chennai 600035, Tamil Nadu (India)

    2013-05-15

    Highlights: ► Magnetic nanoparticles were synthesized by chemical co-precipitation method. ► Surface was functionalized with amino-silane and used for lipase immobilization. ► Characterized through TEM, SEM, XRD, FT-IR and VSM analysis. ► The functionalization and immobilization did not affect the magnetite properties. ► The immobilized lipase showed greater functional property than free lipase. - Abstract: Superparamagnetic nanoparticles (Fe{sub 3}O{sub 4}–magnetite) were prepared by chemical co-precipitation method and their surface was functionalized with 3-aminopropyltriethoxysilane via silanization reaction to obtain amino functionalized magnetic nanoparticles. The purified lipase from Pseudomonas fluorescens Lp1 was immobilized onto functionalized magnetite using glutaraldehyde as the coupling agent. The characterization of the nanoparticles was done by scanning electron microscopy, transmission electron microscopy, powder X-ray diffraction, vibrating sample magnetometry and Fourier transformed infrared spectroscopy. The size of the magnetite was measured about 10–30 nm. The results of characterization study revealed the successful immobilization of lipase on to functionalized magnetite. The saturation magnetization of magnetic nanoparticles was found to be 28.34 emu/g whereas the immobilized magnetic nanoparticle was 17.074 emu/g. The immobilized lipase had greater activity at 50 °C and thermal stability upto 70 °C. It exhibited excellent reusability for 4 cycles and storage stability upto 15 days by retaining 75% of its initial activity.

  17. Efficacy of Pseudomonas fluorescens strain CL145A spray dried powder for controlling zebra mussels adhering to native unionid mussels within field enclosures

    Science.gov (United States)

    Luoma, James A.; Weber, Kerry L.; Severson, Todd J.; Mayer, Denise A.

    2015-01-01

    The efficacy of a commercially prepared spray dried powder (SDP) formulation of Pseudomonas fluorescens (strain CL145A) was evaluated for removing zebra mussels (Dreissena polymorpha) adhering to a population of unionid mussels in Lake Darling (Alexandria, Minnesota). Two groups of unionid mussels were used in the study. Unionid mussels were collected near the test area, weighed, photographed, individually tagged, and randomly allocated to one of nine test enclosures in equal proportions and then divided into two groups. The first group of unionid mussels (Group 1, n = 5 per test enclosure) were indiscriminately selected from each test enclosure and used to estimate the number of zebra mussels adhering to unionid mussels prior to exposure. The second group of unionid mussels (Group 2, n = 22 per test enclosure) were used to evaluate the efficacy of SDP for removal of adhering zebra mussels. Both Group 1 and Group 2 mussels were used to evaluate the effects of SDP exposure on unionid mussel survival.

  18. Immobilization of Pseudomonas fluorescens lipase on hydrophobic supports and application in biodiesel synthesis by transesterification of vegetable oils in solvent-free systems.

    Science.gov (United States)

    Lima, Lionete N; Oliveira, Gladson C; Rojas, Mayerlenis J; Castro, Heizir F; Da Rós, Patrícia C M; Mendes, Adriano A; Giordano, Raquel L C; Tardioli, Paulo W

    2015-04-01

    This work describes the preparation of biocatalysts for ethanolysis of soybean and babassu oils in solvent-free systems. Polystyrene, Amberlite (XAD-7HP), and octyl-silica were tested as supports for the immobilization of Pseudomonas fluorescens lipase (PFL). The use of octyl-silica resulted in a biocatalyst with high values of hydrolytic activity (650.0 ± 15.5 IU/g), immobilization yield (91.3 ± 0.3 %), and recovered activity (82.1 ± 1.5 %). PFL immobilized on octyl-silica was around 12-fold more stable than soluble PFL, at 45 °C and pH 8.0, in the presence of ethanol at 36 % (v/v). The biocatalyst provided high vegetable oil transesterification yields of around 97.5 % after 24 h of reaction using babassu oil and around 80 % after 48 h of reaction using soybean oil. The PFL-octyl-silica biocatalyst retained around 90 % of its initial activity after five cycles of transesterification of soybean oil. Octyl-silica is a promising support that can be used to immobilize PFL for subsequent application in biodiesel synthesis.

  19. Getting the ecology into the interactions between plants and the plant-growth promoting bacterium Pseudomonas fluorescens

    NARCIS (Netherlands)

    Hol, W.H.G.; Bezemer, T.M.; Biere, A.

    2013-01-01

    Plant growth-promoting rhizobacteria (PGPR) are increasingly appreciated for their contributions to primary productivity through promotion of growth and triggering of induced systemic resistance in plants. Here we focus on the beneficial effects of one particular species of PGPR (Pseudomonas

  20. Bacteriophage-derived enzyme that depolymerizes the alginic acid capsule associated with cystic fibrosis isolates of Pseudomonas aeruginosa.

    Science.gov (United States)

    Glonti, T; Chanishvili, N; Taylor, P W

    2010-02-01

    To identify enzymes associated with bacteriophages infecting cystic fibrosis (CF) strains of Pseudomonas aeruginosa that are able to degrade extracellular alginic acids elaborated by the host bacterium. Plaques produced by 21 Ps. aeruginosa-specific phages were screened for the presence of haloes, an indicator of capsule hydrolytic activity. Four phages produced haloed plaques, and one (PT-6) was investigated further. PT-6 was shown by electron microscopy to belong to Podoviridae family C1, to reduce the viscosity of four alginate preparations using a rolling ball viscometer and to release uronic acid-containing fragments from the polymers, as judged by spectrophotometry and thin layer chromatography. The alginase was partially purified by gel filtration chromatography and shown to be a 37 kDa polypeptide. Infection of CF strains of Ps. aeruginosa by phage PT-6 involves hydrolysis of the exopolysaccharide secreted by the host. The alginase produced by PT-6 has the potential to increase the well-being of CF suffers by improving the surface properties of sputum, accelerating phagocytic uptake of bacteria and perturbing bacterial growth in biofilms.

  1. Evidence for a role of biosurfactants produced by Pseudomonas fluorescens in the spoilage of fresh aerobically stored chicken meat.

    Science.gov (United States)

    Mellor, Glen E; Bentley, Jessica A; Dykes, Gary A

    2011-08-01

    Fresh chicken meat is a fat-rich environment and we therefore hypothesised that production of biosurfactants to increase bioavailability of fats may represent one way in which spoilage bacteria might enhance the availability of nutrients. Numbers of Pseudomonas were determined on a total of 20 fresh and 20 spoiled chicken thighs with skin. A total of 400 randomly isolated Pseudomonas colonies from fresh (200) and spoiled (200) chicken were screened for the presence of biosurfactant production. Biosurfactant producing strains represented 5% and 72% of the Pseudomonas spp. isolates from fresh (mean count 2.3 log(10) cfu g(-1)) and spoiled (mean count 7.4 log(10) cfu g(-1)) chicken skin, respectively. Partially-purified biosurfactants derived from a subgroup of four Pseudomonasfluorescens strains obtained through the screening process were subsequently used to investigate the role that the addition of these compounds plays in the spoilage of aerobically stored chicken. Emulsification potential of the four selected biosurfactants was measured against a range of hydrocarbons and oils. All four biosurfactants displayed a greater ability to emulsify rendered chicken fat than hydrocarbons (paraffin liquid, toluene and hexane) and oils (canola, olive, sunflower and vegetable). Storage trials (4 °C) of chicken meat treated with the four selected biosurfactants revealed a significantly greater (P increase in total aerobic count (1.3-1.7 log(10) cfu g(-1)) occurred following one day of incubation. These results indicate that biosurfactants produced by Pseudomonas spp. may play an important role in the spoilage of aerobically stored chicken meat by making nutrients more freely available and providing strains producing them with a competitive advantage. Copyright © 2011 Elsevier Ltd. All rights reserved.

  2. Adaptive Divergence in Experimental Populations of Pseudomonas fluorescens. IV. Genetic Constraints Guide Evolutionary Trajectories in a Parallel Adaptive Radiation

    OpenAIRE

    McDonald, Michael J.; Gehrig, Stefanie M.; Meintjes, Peter L.; Zhang, Xue-Xian; Rainey, Paul B.

    2009-01-01

    The capacity for phenotypic evolution is dependent upon complex webs of functional interactions that connect genotype and phenotype. Wrinkly spreader (WS) genotypes arise repeatedly during the course of a model Pseudomonas adaptive radiation. Previous work showed that the evolution of WS variation was explained in part by spontaneous mutations in wspF, a component of the Wsp-signaling module, but also drew attention to the existence of unknown mutational causes. Here, we identify two new muta...

  3. Membrane-anchored MucR mediates nitrate-dependent regulation of alginate production in Pseudomonas aeruginosa

    KAUST Repository

    Wang, Yajie; Hay, Iain D.; Rehman, Zahid Ur; Rehm, Bernd H A

    2015-01-01

    of MucR impaired alginate promoter activity and global c-di-GMP levels, alginate yields were not directly correlated with alginate promoter activity or c-di-GMP levels, suggesting that nitrate and MucR modulate alginate production at a post

  4. Enhanced biosorption of mercury(II) and cadmium(II) by cold-induced hydrophobic exobiopolymer secreted from the psychrotroph Pseudomonas fluorescens BM07

    Energy Technology Data Exchange (ETDEWEB)

    Zamil, Sheikh Shawkat; Choi, Mun Hwan; Song, Jung Hyun; Park, Hyunju; Xu, Ju; Yoon, Sung Chul [Gyeongsang National Univ., Jinju (Korea). Nano-Biomaterials Science Lab.; Chi, Ki-Whan [Ulsan Univ. (Korea). Dept. of Chemistry

    2008-09-15

    The cells of psychrotrophic Pseudomonas fluorescens BM07 were found to secrete large amounts of exobiopolymer (EBP) composed of mainly hydrophobic (water insoluble) polypeptide(s) (as contain {proportional_to}50 mol% hydrophobic amino acids, lacking cysteine residue) when grown on fructose containing limited M1 medium at the temperatures as low as 0-10 C but trace amount at high (30 C, optimum growth) temperature. Two types of nonliving BM07 cells (i.e., cells grown at 30 C and 10 C) as well as the freeze-dried EBP were compared for biosorption of mercury (Hg(II)) and cadmium (Cd(II)). The optimum adsorption pH was found 7 for Hg(II) but 6 for Cd(II), irrespective of the type of biomass. Equilibrium adsorption data well fitted the Langmuir adsorption model. The maximum adsorption (Q{sub max}) was 72.3, 97.4, and 286.2 mg Hg(II)/g dry biomass and 18.9, 27.0, and 61.5 mg Cd(II)/g dry biomass for cells grown at 30 C and 10 C and EBP, respectively, indicating major contribution of heavy metal adsorption by cold-induced EBP. Mercury(II) binding induced a significant shift of infrared (IR) amide I and II absorption of EBP whereas cadmium(II) binding showed only a very little shift. These IR shifts demonstrate that mercury(II) and cadmium(II) might have different binding sites in EBP, which was supported by X-ray diffraction and differential scanning calorimetric analysis and sorption results of chemically modified biomasses. This study implies that the psychrotrophs like BM07 strain may play an important role in the bioremediation of heavy metals in the temperate regions especially in the inactive cold season. (orig.)

  5. Prokaryotic Homologs of the Eukaryotic 3-Hydroxyanthranilate 3,4-Dioxygenase and 2-Amino-3-Carboxymuconate-6-Semialdehyde Decarboxylase in the 2-Nitrobenzoate Degradation Pathway of Pseudomonas fluorescens Strain KU-7†

    OpenAIRE

    Muraki, Takamichi; Taki, Masami; Hasegawa, Yoshie; Iwaki, Hiroaki; Lau, Peter C. K.

    2003-01-01

    The 2-nitrobenzoic acid degradation pathway of Pseudomonas fluorescens strain KU-7 proceeds via a novel 3-hydroxyanthranilate intermediate. In this study, we cloned and sequenced a 19-kb DNA locus of strain KU-7 that encompasses the 3-hydroxyanthranilate meta-cleavage pathway genes. The gene cluster, designated nbaEXHJIGFCDR, is organized tightly and in the same direction. The nbaC and nbaD gene products were found to be novel homologs of the eukaryotic 3-hydroxyanthranilate 3,4-dioxygenase a...

  6. Salicylic acid degradation from aqueous solutions using Pseudomonas fluorescens HK44: parameters studies and application tools Degradação de ácido salicílico presente em soluções sintéticas utilizando Pseudomonas fluorescens HK44

    Directory of Open Access Journals (Sweden)

    Tatyane R. Silva

    2007-03-01

    Full Text Available The optimal conditions for salicylic acid biodegradation by Pseudomonas fluorescens HK44 were determined in this study with the intention to create a microbial sensor. Kinetic experiments permitted a definition of 60 and 30min the time needed to achieve the maximum degradation of salicylic acid presented in a medium with and without yeast extract, respectively. The degradation in medium without yeast extract and the quantification by spectrophotometry 230 nm were selected to be used in further tests. The use of preactivated cells or on the exponential growth phase showed better salicylic acid degradation percentages when compared to nonactivated cells or on the stationary growth state. Finally, the best cellular concentration used on the salicylic acid degradation was 0,1 g.L-1. Strain HK44 shows to be capable of degrade salicylic acid presented in simple aqueous systems, making this strain a promising tool for the application on a luminescent microbial sensor.Com a intenção de criar um sensor microbiano, as condições ótimas para a biodegradação de ácido salicílico por Pseudomonas fluorescens HK44 foram determinadas neste estudo. Os experimentos cinéticos permitiram a definição dos tempos de 60 e 30 minutos como necessários para atingir a máxima degradação de ácido salicílico presente em meio com ou sem extrato de lêvedo, respectivamente. A degradação no meio sem extrato de lêvedo e a quantificação através de espectrofotometria 230 nm foram selecionadas para serem utilizadas em testes posteriores. O uso de células pré-ativadas ou na fase exponencial de crescimento apresentou melhores porcentagens de degradação de ácido salicílico quando comparadas a células não-ativadas ou no estado estacionário de crescimento. Além disso, a melhor concentração celular utilizada nessa degradação foi 0,1 g.L¹. A cepa HK44 parece ser capaz de degradar o ácido salicílico presente em sistemas aquosos simples, tornando este

  7. From the root to the stem: interaction between the biocontrol root endophyte Pseudomonas fluorescens PICF7 and the pathogen Pseudomonas savastanoi NCPPB 3335 in olive knots

    Science.gov (United States)

    Maldonado-González, M Mercedes; Prieto, Pilar; Ramos, Cayo; Mercado-Blanco, Jesús

    2013-01-01

    Olive knot disease, caused by Pseudomonas savastanoi pv. savastanoi, is one of the most important biotic constraints for olive cultivation. Pseudomonas fluorescens PICF7, a natural colonizer of olive roots and effective biological control agent (BCA) against Verticillium wilt of olive, was examined as potential BCA against olive knot disease. Bioassays using in vitro-propagated olive plants were carried out to assess whether strain PICF7 controlled knot development either when co-inoculated with the pathogen in stems or when the BCA (in roots) and the pathogen (in stems) were spatially separated. Results showed that PICF7 was able to establish and persist in stem tissues upon artificial inoculation. While PICF7 was not able to suppress disease development, its presence transiently decreased pathogen population size, produced less necrotic tumours, and sharply altered the localization of the pathogen in the hyperplasic tissue, which may pose epidemiological consequences. Confocal laser scanning microscopy combined with fluorescent tagging of bacteria revealed that when PICF7 was absent the pathogen tended to be localized at the knot surface. However, presence of the BCA seemed to confine P. savastanoi at inner regions of the tumours. This approach has also enabled to prove that the pathogen can moved systemically beyond the hypertrophied tissue. PMID:23425069

  8. INFLUENCIA DE LA BACTERIA Pseudomonas fluorescens EN LA LECHE, SOBRE LAS CARACTERÍSTICAS SENSORIALES DEL QUESO DOBLE CREMA

    Directory of Open Access Journals (Sweden)

    SJ González

    2007-01-01

    Full Text Available El objetivo de este trabajo fue observar el efecto de la presencia de microorganismos psicro-tróÀcos en la leche, sobre las características sensoriales del queso doble crema. Para esto, se inoculó la leche con un cultivo de Pseudomonas Áuorescens, a un nivel de 108 unidades formadoras de colonia por mililitro (ufc/ml. La leche se refrigeró a 4 ºC por seis días, se pasteurizó y posteriormente se preparó el queso. Se elaboraron lotes de queso cada 10 días, con el Àn de obtener siete tiempos experimentales. A cada uno se le determinó la vida útil sensorial. Los resultados de las pruebas sensoriales se analizaron con el programa S-PLUS de estadísti-ca de supervivencia aplicada a la vida útil sensorial de alimentos. Se determinó que el tiempo de vida útil sensorial del queso doble crema elaborado con leche en estas condiciones, y alma-cenado a 4 °C, está alrededor de 30 días, disminuyendo en un 50% con relación al patrón.

  9. Regulation of Soluble Phosphate on the Ability of Phytate Mineralization and β-Propeller Phytase Gene Expression of Pseudomonas fluorescens JZ-DZ1, a Phytate-Mineralizing Rhizobacterium.

    Science.gov (United States)

    Shen, Lan; Wu, Xiao-Qin; Zeng, Qing-Wei; Liu, Hong-Bin

    2016-12-01

    Phytate-mineralizing rhizobacteria (PMR) play an important role in providing phosphorus for the sustainable plant growth. It is important to investigate the ability of PMR to produce phytase under different phosphate levels for its application. The effects of different concentrations of soluble phosphate on the ability of phytate mineralization of Pseudomonas fluorescens JZ-DZ1, a phytate-mineralizing rhizobacterium, were investigated in both solid and liquid media. The results on solid media showed that halo zone width gradually reduced with concentrations of soluble phosphate increasing from 0.05 to 20 mM, indicating the reduction of the ability of phytate mineralization. The results were consistent with the quantitative detection of phytase activity from the overall trend. An 1866-bp β-propeller phytase (BPP) gene (phyPf) was cloned from the strain, and the deduced amino acid sequence of phyPf shared 98 % of identity with a known BPP from Pseudomonas sp. BS10-3 (AJF36073.1). The results of relative real-time quantitative PCR assay showed that the expression of phyPf was induced by a low concentration (0.1 mM) of soluble phosphate, suggesting that BPP secretion was regulated by gene phyPf. The BPP-harboring bacterium P. fluorescens JZ-DZ1 with low phosphate-inducible ability of phytate mineralization could be potentially applied to promote phosphorus uptake for plants in the future.

  10. Preparation and characterization of monodisperse microcapsules with alginate and bentonite via external gelation technique encapsulating Pseudomonas putida Rs-198.

    Science.gov (United States)

    Li, Xuan; Wu, Zhansheng; He, Yanhui; Ye, Bang-Ce; Wang, Jun

    2017-10-01

    This paper evaluated the external gelation technique for preparing microcapsules. The microcapsules were consisted of Pseudomonas putida Rs-198 (Rs-198) core and sodium alginate (NaAlg)-bentonite (Bent) shell. Different emulsification rotation speeds and core/shell ratios were used to prepare the microcapsules of each formulation. The near-spherical microcapsules were monodisperse with a mean diameter of 25-100 μm and wrinkled surfaces. Fourier transform infrared spectrophotometry (FTIR) and thermogravimetric analysis (TGA) revealed the physical mixture of the wall material and the superior thermal stability of the microcapsules. Percentage yield, water content, and encapsulation efficiency were evaluated and correlated with the changes in emulsification rotation speed and core/shell ratio. In vitro release experiments demonstrated that 60% of the bacteria were released from the NaAlg-Bent microcapsules within three days. Considerably better survival was observed for encapsulated cells compared to free cells, especially in pH 4.0 and 10.0. In summary, the desired properties of microcapsules can be obtained by external gelation technique and the microcapsules on the bacteria had a good protective effect.

  11. The interplay of StyR and IHF regulates substrate-dependent induction and carbon catabolite repression of styrene catabolism genes in Pseudomonas fluorescens ST

    Directory of Open Access Journals (Sweden)

    Leoni Livia

    2008-06-01

    Full Text Available Abstract Background In Pseudomonas fluorescens ST, the promoter of the styrene catabolic operon, PstyA, is induced by styrene and is subject to catabolite repression. PstyA regulation relies on the StyS/StyR two-component system and on the IHF global regulator. The phosphorylated response regulator StyR (StyR-P activates PstyA in inducing conditions when it binds to the high-affinity site STY2, located about -40 bp from the transcription start point. A cis-acting element upstream of STY2, named URE, contains a low-affinity StyR-P binding site (STY1, overlapping the IHF binding site. Deletion of the URE led to a decrease of promoter activity in inducing conditions and to a partial release of catabolite repression. This study was undertaken to assess the relative role played by IHF and StyR-P on the URE, and to clarify if PstyA catabolite repression could rely on the interplay of these regulators. Results StyR-P and IHF compete for binding to the URE region. PstyA full activity in inducing conditions is achieved when StyR-P and IHF bind to site STY2 and to the URE, respectively. Under catabolite repression conditions, StyR-P binds the STY1 site, replacing IHF at the URE region. StyR-P bound to both STY1 and STY2 sites oligomerizes, likely promoting the formation of a DNA loop that closes the promoter in a repressed conformation. We found that StyR and IHF protein levels did not change in catabolite repression conditions, implying that PstyA repression is achieved through an increase in the StyR-P/StyR ratio. Conclusion We propose a model according to which the activity of the PstyA promoter is determined by conformational changes. An open conformation is operative in inducing conditions when StyR-P is bound to STY2 site and IHF to the URE. Under catabolite repression conditions StyR-P cellular levels would increase, displacing IHF from the URE and closing the promoter in a repressed conformation. The balance between the open and the closed

  12. A complex multilevel attack on Pseudomonas aeruginosa algT/U expression and AlgT/U activity results in the loss of alginate production

    DEFF Research Database (Denmark)

    Sautter, Robert; Ramos, Damaris; Schneper, Lisa

    2012-01-01

    Infection by the opportunistic pathogen Pseudomonas aeruginosa is a leading cause of morbidity and mortality seen in cystic fibrosis (CF) patients. This is mainly due to the genotypic and phenotypic changes of the bacteria that cause conversion from a typical nonmucoid to a mucoid form in the CF...... lung. Mucoid conversion is indicative of overproduction of a capsule-like polysaccharide called alginate. The alginate-overproducing (Alg+) mucoid phenotype seen in the CF isolates is extremely unstable. Low oxygen tension growth of mucoid variants readily selects for nonmucoid variants. The switching...... complementation and sequencing of one Group B sap mutant, sap22, revealed that the nonmucoid phenotype was due to the presence of a mutation in PA3257. PA3257 encodes a putative periplasmic protease. Mutation of PA3257 resulted in decreased algT/U expression. Thus, inhibition of algT/U is a primary mechanism...

  13. Kinetics of biofilm formation and desiccation survival of Listeria monocytogenes in single and dual species biofilms with Pseudomonas fluorescens, Serratia proteamaculans or Shewanella baltica on food-grade stainless steel surfaces.

    Science.gov (United States)

    Daneshvar Alavi, Hessam Edin; Truelstrup Hansen, Lisbeth

    2013-01-01

    This study investigated the dynamics of static biofilm formation (100% RH, 15 °C, 48-72 h) and desiccation survival (43% RH, 15 °C, 21 days) of Listeria monocytogenes, in dual species biofilms with the common spoilage bacteria, Pseudomonas fluorescens, Serratia proteamaculans and Shewanella baltica, on the surface of food grade stainless steel. The Gram-negative bacteria reduced the maximum biofilm population of L. monocytogenes in dual species biofilms and increased its inactivation during desiccation. However, due to the higher desiccation resistance of Listeria relative to P. fluorescens and S. baltica, the pathogen survived in greater final numbers. In contrast, S. proteamaculans outcompeted the pathogen during the biofilm formation and exhibited similar desiccation survival, causing the N21 days of Serratia to be ca 3 Log10(CFU cm(-2)) greater than that of Listeria in the dual species biofilm. Microscopy revealed biofilm morphologies with variable amounts of exopolymeric substance and the presence of separate microcolonies. Under these simulated food plant conditions, the fate of L. monocytogenes during formation of mixed biofilms and desiccation depended on the implicit characteristics of the co-cultured bacterium.

  14. Quantification of Pseudomonas fluorescens strains F113, CHA0 and Pf153 in the rhizosphere of maize by strain-specific real-time PCR unaffected by the variability of DNA extraction efficiency.

    Science.gov (United States)

    Von Felten, Andreas; Défago, Geneviève; Maurhofer, Monika

    2010-05-01

    Pseudomonas fluorescens strains F113 and CHA0 are well-known plant growth-promoting rhizobacteria (PGPR) often used as model strains in biocontrol experiments. To monitor their persistence in large scale field experiments, culture-independent methods are needed. In this study, a strain-specific real-time PCR quantification tool was developed based on sequence-characterized amplified regions (SCAR) for P. fluorescens strains F113, CHA0 and Pf153. Differences in DNA extraction efficiencies from rhizosphere samples were circumvented using plasmid APA9 as internal standard to normalize C(T) values after real-time amplification. The detection limits of the real-time PCR assays for all three strains were approximately 10 cells for genomic DNA and 10(4)cells/g rhizosphere for maize samples grown in different natural soils. Population sizes of the three strains in the rhizosphere of maize measured by the new real-time PCR approaches were similar to those measured by most probable number (MPN)-PCR. A persistence study of the three strains indicated that the strains persisted differently over a period of 5weeks. In conclusion the newly developed real-time PCR approach is a fast and resource efficient method for monitoring individual biocontrol strains in natural soil, which makes it an apt quantification tool for future large-scale field experiments. Copyright 2010 Elsevier B.V. All rights reserved.

  15. Characterization of the CrbS/R Two-Component System in Pseudomonas fluorescens Reveals a New Set of Genes under Its Control and a DNA Motif Required for CrbR-Mediated Transcriptional Activation

    Directory of Open Access Journals (Sweden)

    Edgardo Sepulveda

    2017-11-01

    Full Text Available The CrbS/R system is a two-component signal transduction system that regulates acetate utilization in Vibrio cholerae, P. aeruginosa, and P. entomophila. CrbS is a hybrid histidine kinase that belongs to a recently identified family, in which the signaling domain is fused to an SLC5 solute symporter domain through aSTAC domain. Upon activation by CrbS, CrbR activates transcription of the acs gene, which encodes an acetyl-CoA synthase (ACS, and the actP gene, which encodes an acetate/solute symporter. In this work, we characterized the CrbS/R system in Pseudomonas fluorescens SBW25. Through the quantitative proteome analysis of different mutants, we were able to identify a new set of genes under its control, which play an important role during growth on acetate. These results led us to the identification of a conserved DNA motif in the putative promoter region of acetate-utilization genes in the Gammaproteobacteria that is essential for the CrbR-mediated transcriptional activation of genes under acetate-utilizing conditions. Finally, we took advantage of the existence of a second SLC5-containing two-component signal transduction system in P. fluorescens, CbrA/B, to demonstrate that the activation of the response regulator by the histidine kinase is not dependent on substrate transport through the SLC5 domain.

  16. Non-target effects of the microbial control agents Pseudomonas fluorescens DR54 and Clonostachys rosea IK726 in soils cropped with barley followed by sugar beet: a greenhouse assessment

    DEFF Research Database (Denmark)

    Johansen, A.; Knudsen, I. M. B.; Binnerup, S. J.

    2005-01-01

    Non-target effects of a bacterial (Pseudomonas fluorescens DR54) and a fungal (Clonostachys rosea IK726) microbial control agent (MCA), on the indigenous microbiota in bulk soil and rhizosphere of barley, and subsequent a sugar beet crop, were studied in a greenhouse experiment. MCAs were...... introduced by seed and soil inoculation. Bulk and rhizosphere soils were sampled regularly during the growth of barley and sugar beet. The soils were assayed for the fate of MCAs and various features of the indigenous soil microbiota. At the end of the experiment (193 d), DR54 and IK726 had declined...... by a factor of 106 and 20, respectively, and DR54 showed a short-lasting growth increase in the sugar beet rhizosphere. In general, the non-target effects were small and transient. IK726 seemed to have general stimulating effects on soil enzyme activity and the soil microbiota, and resulted in a significant...

  17. INOCULACIÓN AL SUELO CON Pseudomonas fluorescens, Azospirillum oryzae, Bacillus subtilis Y MICROORGANISMOS DE MONTAÑA (MM Y SU EFECTO SOBRE UN SISTEMA DE ROTACIÓN SOYA-TOMATE BAJO CONDICIONES DE INVERNADERO

    Directory of Open Access Journals (Sweden)

    Leida Castro Barquero

    2015-01-01

    Full Text Available Se evaluó un sistema de rotación soya- tomate, con incorporación de biomasa verde y aplicación de inóculos microbianos individuales y en mezcla sobre el crecimiento de las plantas y propiedades edáficas; para ello se evaluaron en invernadero por 24 meses los siguientes 9 trata - mientos: solo tomate (T; rotación tomate-soya (TS; rotación tomate-soya con inoculaciones individuales de Azospirillum oryzae (A; de Pseudomonas fluorescens (P; de Bacillus sub - tilis (B; de microorganismos de montaña (MM; y las inoculaciones en mezcla de B. subtilis y P. fluorescens (BP; de B. subtilis, P. fluorescens y A. oryzae ( BPA; d e B. subtilis , P. fluorescens , Azospirillum sp. y MM (BPAMM. Se evalua - ron las variables físicas: densidad aparente y de partículas; conductividad hidráulica; poros totales; estabilidad de agregados; resistencia a la penetración (RP; las variables químicas: pH; conductividad eléctrica; contenido de N y C; relación C/N; contenido de nutrientes en suelos y foliares; las variables biológicas: respiración de suelos, unidades formadoras de colonias de hongos, bacterias y actinomicetos y el peso fres - co y seco foliar. Las variables físicas no fueron afectadas por los tratamientos, con excepción de RP, que fue mayor en el tratamiento T. Las varia - bles biológicas y químicas fueron sensibles a los tratamientos, con valores significativamente más altos en presencia de MM. Destaca el incremento del P en solución de suelo en tratamientos a los que se adicionó MM: pasó de 6 a 20 mg.l -1 ; esta condición se reflejó además en la cantidad de P en el tejido foliar al final del ensayo. Se determi - nó que el pH, CE y la respiración del suelo fueron afectados por la interacción entre los tratamientos aplicados y el tiempo transcurrido; los mayores valores se obtuvieron al final del ensayo y en los tratamientos con MM.

  18. Inoculación al suelo con Pseudomonas fluorescens, Azospirillum oryzae, Bacillus subtilis y microorganismos de montaña (mm y su efecto sobre un sistema de rotación soya-tomate bajo condiciones de invernadero

    Directory of Open Access Journals (Sweden)

    Leida Castro Barquero

    2015-11-01

    Full Text Available Se evaluó un sistema de rotación soyatomate, con incorporación de biomasa verde y aplicación de inóculos microbianos individuales y en mezcla sobre el crecimiento de las plantas y propiedades edáficas; para ello se evaluaron en invernadero por 24 meses los siguientes 9 tratamientos: solo tomate (T; rotación tomate-soya (TS; rotación tomate-soya con inoculaciones individuales de Azospirillum oryzae (A; de Pseudomonas fluorescens (P; de Bacillus subtilis (B; de microorganismos de montaña (MM; y las inoculaciones en mezcla de B. subtilis y P. fluorescens (BP; de B. subtilis, P. fluorescens y A. oryzae (BPA; de B. subtilis, P. fluorescens, Azospirillum sp. y MM (BPAMM. Se evaluaron las variables físicas: densidad aparente y de partículas; conductividad hidráulica; poros totales; estabilidad de agregados; resistencia a la penetración (RP; las variables químicas: pH; conductividad eléctrica; contenido de N y C; relación C/N; contenido de nutrientes en suelos y foliares; las variables biológicas: respiración de suelos, unidades formadoras de colonias de hongos, bacterias y actinomicetos y el peso fresco y seco foliar. Las variables físicas no fueron afectadas por los tratamientos, con excepción de RP, que fue mayor en el tratamiento T. Las variables biológicas y químicas fueron sensibles a los tratamientos, con valores significativamente más altos en presencia de MM. Destaca el incremento del P en solución de suelo en tratamientos a los que se adicionó MM: pasó de 6 a 20 mg.l-1; esta condición se reflejó además en la cantidad de P en el tejido foliar al final del ensayo. Se determinó que el pH, CE y la respiración del suelo fueron afectados por la interacción entre los tratamientos aplicados y el tiempo transcurrido; los mayores valores se obtuvieron al final del ensayo y en los tratamientos con MM.

  19. Control of Fusarium verticillioides, cause of ear rot of maize, by Pseudomonas fluorescens

    DEFF Research Database (Denmark)

    Nayaka, Siddaiah Chandra; Shankar, Akarere C. Udaya; Reddy, Munagala S.

    2009-01-01

    Abstract BACKGROUND: Maize is one of the staple food crops grown in India. Fusarium verticillioides (Sacc.) Nirenberg is the most important fungal pathogen of maize, associated with diseases such as ear rot and kernel rot. Apart from the disease, it is capable of producing fumonisins, which have...... disease and fumonisin accumulation, and also to study the capacity to promote growth and yield of maize. In vitro assays were conducted to test the efficacy of P. fluorescens as a seed treatment on seed germination, seedling vigour and also the incidence of F. verticillioides in different maize cultivars....... verticillioides and the level of fumonisins to a maximum extent compared with the other treatments. CONCLUSION: The study demonstrates the potential role of P. fluorescens and its formulations in ear rot disease management. The biocontrol potential of this isolate is more suited for fumonisin reduction in maize...

  20. Phenylacetic acid-producing Rhizoctonia solani represses the biosynthesis of nematicidal compounds in vitro and influences biocontrol of Meloidogyne incognita in tomato by Pseudomonas fluorescens strain CHA0 and its GM derivatives.

    Science.gov (United States)

    Siddiqui, I A; Shaukat, S S

    2005-01-01

    The aim of the present investigation was to determine the influence of Rhizoctonia solani and its pathogenicity factor on the production of nematicidal agent(s) by Pseudomonas fluorescens strain CHA0 and its GM derivatives in vitro and nematode biocontrol potential by bacterial inoculants in tomato. One (Rs7) of the nine R. solani isolates from infected tomato roots inhibited seedling emergence and caused root rot in tomato. Thin layer chromatography revealed that culture filtrates of two isolates (Rs3 and Rs7) produced brown spots at Rf-values closely similar to synthetic phenylacetic acid (PAA), a phytotoxic factor. Filtrates from isolate Rs7, amended with the growth medium of P. fluorescens, markedly repressed nematicidal activity and PhlA'-'LacZ reporter gene expression of the bacteria in vitro. On the contrary, isolate Rs4 enhanced nematicidal potential of a 2,4-diacetylphloroglucinol overproducing mutant, CHA0/pME3424, of P. fluorescens strain CHA0 in vitro. Therefore, R. solani isolates Rs4 and Rs7 were tested more rigorously for their potential to influence biocontrol effectiveness of the bacterial agents. Methanol extract of the culture filtrates of PAA-producing isolate Rs7 resulting from medium amended with phenylalanine enhanced fungal repression of the production of nematicidal agents by bacteria, while amendments with zinc or molybdenum eliminated such fungal repression, thereby restoring bacterial potential to cause nematode mortality in vitro. A pot experiment was carried out, 3-week-old tomato seedlings were infested with R. solani isolates Rs4 or Rs7 and/or inoculated with Meloidogyne incognita, the root-knot nematode. The infested soil was treated with aqueous cell suspensions (10(8) CFU) of P. fluorescens strain CHA0 or its GM derivatives or left untreated (as a control). Observations taken 45 days after nematode inoculation revealed that, irrespective of the bacterial treatments, galling intensity per gram of fresh tomato roots was markedly

  1. Análisis de las polihidroxialcanoato sintasas (PhaC1 y PhaC2 en una cepa de Pseudomonas fluorescens IBUN S1602, aislada en suelos colombianos

    Directory of Open Access Journals (Sweden)

    Julieth Serrano Riaño

    2011-07-01

    Full Text Available Título en ingles: Analysis of polyhydroxyalkanoate synthases (PhaC1 and PhaC2 in a strain of Pseudomonas fluorescens   IBUN S1602 isolated from Colombian soil. Resumen La cepa Pseudomonas fluorescens IBUN S1602 conforma el grupo de aislamientos provenientes de suelos colombianos de caña de azúcar, que acumula polihidrioxialcanoato (PHA, fue seleccionada como promisoria para escalamiento comercial por tener afinidad por sustratos alternativos y económicos como el glicerol, aceites usados, suero de leche, entre otros. Dada la importancia de la enzima sintasa en la síntesis de los PHAs, en el presente trabajo se realizó el análisis molecular de los genes phaC1 y phaC2 que codifican las enzimas sintasas tipo II (PhaC1 y PhaC2. Para la obtención de los amplímeros requeridos en la secuenciación, se utilizó la técnica de PCR bajo condiciones estandarizadas para iniciadores diseñados reportados en las bases de datos. Se identificaron dos fragmentos de 1680 pb y 1683 pb correspondientes a  phaC1 y phaC2. El análisis comparativo de las secuencias proteicas resultantes de estos genes demuestra que la sintasa  IBUN S1602 contiene la región α/β hidrolasa y 8 residuos de aminoácidos conservados, que son características de las sintasas examinadas a nivel mundial. Se analizó la estructura enzimática a nivel primario y se predijo la secundaria. Se concluyó que  las sintasas de la cepa Pseudomonas fluorescens IBUN S1602 presentan alta homología con las sintasas tipo II  que se reportan para Pseudomonas. Los resultados obtenidos contribuyen al entendimiento básico de la biosíntesis de PHA,  la cual permitirá, en un futuro, el aumento de la calidad de PHA debida a la modulación del nivel de sintasa que se exprese en un organismo recombinante, con el fin de variar el peso molecular del biopolímero, propiedad esencial en el estudio de aplicaciones industriales.   Palabras clave: polihidroxialcanoatos, PHA sintasa, bioinform

  2. Evaluation of gamma irradiation effect and Pseudomonas ...

    African Journals Online (AJOL)

    Antagonistic effect of Pseudomonas fluorescens and influence of gamma irradiation on the development of Penicillium expansum, the causal agent of postharvest disease on apple fruit was studied. P. fluorescens was originally isolated from rhizosphere of the apple trees. Suspension of P. fluorescens and P. expansum ...

  3. Accelerated storage testing of freeze-dried Pseudomonas ...

    African Journals Online (AJOL)

    Accelerated storage testing of freeze-dried Pseudomonas fluorescens BTP1, ... of all P. fluorescens strains were not significantly different and thermal inactivation ... useful to the development of improved reference materials and samples held ...

  4. Lysophosphatidic acid inhibition of the accumulation of Pseudomonas aeruginosa PAO1 alginate, pyoverdin, elastase and LasA

    DEFF Research Database (Denmark)

    Laux, D.C.; Corson, J.M.; Givskov, Michael Christian

    2002-01-01

    . In the present study, a lysophospholipid, 1-paimitoyl-2-hydroxy-sn-glycero-3-phosphate [also called monopalmitoylphosphatidic acid (MPPA)], which accumulates in inflammatory exudates, was shown to inhibit the extracellular accumulation of P. aeruginosa PAO1 alginate, elastase, LasA protease and the siderophore...

  5. Identification of copper-induced genes in Pseudomonas fluorescens and use of a reporter strain to monitor bioavailable copper in soil

    DEFF Research Database (Denmark)

    Tom-Petersen, Andreas; Hosbond, Carsten; Nybroe, Ole

    2001-01-01

    -amended soil microcosms in a concentration-dependent manner. The chelator EDTA reduced the availability of Cu to P. fluorescens in soil. This showed that complex-bound Cu is not necessarily available to bacteria, We compared chemical analysis of soluble Cu and the reporter assay on soil solutions from Cu-containing......-Cu15, the gene interrupted by the transposon encoded a protein carrying a Cu-binding domain but with low homology to known proteins. DF57-Cu15 was the most suitable Cu reporter due to its high specific response and tolerance to Cu in pure culture. DF57-Cu15 responded to soil solutions from Cu...... soil microcosms supplemented with either manure or straw. Organic matter increased the amount of soluble Cu but not the amount of bioavailable Cu. Probably, Cu binds with high affinity to organic constituents in pig manure or barley straw. Hence, determination of soluble Cu by chemical analysis cannot...

  6. Biosynthesis of the antimicrobial cyclic lipopeptides nunamycin and nunapeptin by Pseudomonas fluorescens strain In5 is regulated by the LuxR-type transcriptional regulator NunF

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Phippen, Christopher; Nielsen, Kristian Fog

    2017-01-01

    -producing pseudomonads except for the border regions where putative LuxR-type regulators are located. This study focuses on understanding the regulatory role of the LuxR-type-encoding gene nunF in CLP production of P. fluorescens In5. Functional analysis of nunF coupled with liquid chromatography-high-resolution mass...... spectrometry (LC-HRMS) showed that CLP biosynthesis is regulated by nunF. Quantitative real-time PCR analysis indicated that transcription of the NRPS genes catalyzing CLP production is strongly reduced when nunF is mutated indicating that nunF is part of the nun-nup regulon. Swarming and biofilm formation...... that environmental elicitors may also influence nunF expression which upon activation regulates nunamycin and nunapeptin production required for the growth inhibition of phytopathogens....

  7. Exploiting genotypic diversity of 2,4-Diacetylphloroglucinol-producing Pseudomonas spp.: characterization of superior root-colonizing P. fluorescens strain Q8r1-96

    NARCIS (Netherlands)

    Raaijmakers, J.M.; Weller, D.M.

    2001-01-01

    The genotypic diversity that occurs in natural populations of antagonistic microorganisms provides an enormous resource for improving biological control of plant diseases. In this study, we determined the diversity of indigenous 2,4-diacetylphloroglucinol (DAPG)-producing Pseudomonas spp. occurring

  8. An N-terminal Retention Module Anchors the Giant Adhesin LapA of Pseudomonas fluorescens at the Cell Surface: A Novel Sub-family of Type I Secretion Systems.

    Science.gov (United States)

    Smith, T Jarrod; Font, Maria E; Kelly, Carolyn M; Sondermann, Holger; O'Toole, George A

    2018-02-05

    LapA of Pseudomonas fluorescens Pf0-1 belongs to a diverse family of cell surface associated bacterial adhesins that are secreted via the type-1 secretion system (T1SS). We previously reported that the periplasmic protease LapG cleaves the N-terminus of LapA at a canonical dialanine motif to release the adhesin from the cell surface under conditions unfavorable to biofilm formation, thus decreasing biofilm formation. Here, we characterize LapA as the first type 1 secreted substrate that does not follow the "one-step" rule of T1SS. Rather, a novel N-terminal element, called the retention module (RM), localizes LapA at the cell surface as a secretion intermediate. Our genetic, biochemical, and molecular modeling analysis support a model wherein LapA is tethered to the cell surface through its T1SS outer membrane TolC-like pore, LapE, until LapG cleaves LapA in the periplasm. We further demonstrate this unusual retention strategy is likely conserved among LapA-like proteins, and reveals a new subclass of T1SS ABC transporters involved in transporting this group of surface-associated, LapA-like adhesins. These studies demonstrate a novel cell surface retention strategy used throughout the Proteobacteria and highlight a previously unappreciated flexibility of function for T1SS. Importance. Bacteria have evolved multiple secretion strategies to interact with their environment. For many bacteria, the secretion of cell surface associated adhesins is key for initiating contact with a preferred substratum to facilitate biofilm formation. Our work demonstrates that P. fluorescens uses a previously unrecognized secretion strategy to retain the giant adhesin LapA at its cell surface. Further, we identify likely LapA-like adhesins in various pathogenic and commensal Proteobacteria and provide phylogenetic evidence that these adhesins are secreted by a new subclass of T1SS ABC transporters. Copyright © 2018 American Society for Microbiology.

  9. Complex regulation of AprA metalloprotease in Pseudomonas fluorescens M114: evidence for the involvement of iron, the ECF sigma factor, PbrA and pseudobactin M114 siderophore.

    Science.gov (United States)

    Maunsell, Bláithín; Adams, Claire; O'Gara, Fergal

    2006-01-01

    In the soil bacterium Pseudomonas fluorescens M114, extracellular proteolytic activity and fluorescent siderophore (pseudobactin M114) production were previously shown to be co-ordinately negatively regulated in response to environmental iron levels. An iron-starvation extracytoplasmic function sigma factor, PbrA, required for the transcription of siderophore biosynthetic genes, was also implicated in M114 protease regulation. The current study centred on the characterization and genetic regulation of the gene(s) responsible for protease production in M114. A serralysin-type metalloprotease gene, aprA, was identified and found to encode the major, if not only, extracellular protease produced by this strain. The expression of aprA and its protein product were found to be subject to complex regulation. Transcription analysis confirmed that PbrA was required for full aprA transcription under low iron conditions, while the ferric uptake regulator, Fur, was implicated in aprA repression under high iron conditions. Interestingly, the iron regulation of AprA was dependent on culture conditions, with PbrA-independent AprA-mediated proteolytic activity observed on skim milk agar supplemented with yeast extract, when supplied with iron or purified pseudobactin M114. These effects were not observed on skim milk agar without yeast extract. PbrA-independent aprA expression was also observed from a truncated transcriptional fusion when grown in sucrose asparagine tryptone broth supplied with iron or purified pseudobactin M114. Thus, experimental evidence suggested that iron mediated its effects via transcriptional activation by PbrA under low iron conditions, while an as-yet-unidentified sigma factor(s) may be required for the PbrA-independent aprA expression and AprA proteolytic activity induced by siderophore and iron.

  10. Kynurenine 3-monooxygenase from Pseudomonas fluorescens: substrate-like inhibitors both stimulate flavin reduction and stabilize the flavin-peroxo intermediate yet result in the production of hydrogen peroxide.

    Science.gov (United States)

    Crozier-Reabe, Karen R; Phillips, Robert S; Moran, Graham R

    2008-11-25

    Kynurenine 3-monooxygenase (KMO) is a flavin-dependent hydroxylase that catalyzes the conversion of l-kynurenine (l-Kyn) to 3-hydroxykynurenine (3OHKyn) in the pathway for tryptophan catabolism. KMO inhibition has been widely suggested as an early treatment for stroke and other neurological disorders that involve ischemia. We have investigated the reductive and the oxidative half-reactions of a stable form of KMO from Pseudomonas fluorescens (KMO). The binding of l-Kyn by the enzyme is relatively slow and involves at least two reversible steps. The rate constant for reduction of the flavin cofactor by NADPH increases by a factor of approximately 2.5 x 10(3) when l-Kyn is bound. The rate of reduction of the KMO.l-Kyn complex is 160 s(-1), and the K(d) for the NADPH complex is 200 microM with charge-transfer absorption bands for the KMO(RED).l-Kyn.NADP(+) complex accumulating after reduction. The reduction potential of KMO is -188 mV and is unresponsive to the addition of l-Kyn or other inhibitory ligands. KMO inhibitors whose structures are reminiscent of l-Kyn such as m-nitrobenzoylalanine and benzoylalanine also stimulate reduction of flavin by NADPH and, in the presence of dioxygen, result in the stoichiometric liberation of hydrogen peroxide, diminishing the perceived therapeutic potential of inhibitors of this type. In the presence of the native substrate, the oxidative half-reaction exhibits triphasic absorbance data. A spectrum consistent with that of a peroxyflavin species accumulates and then decays to yield the oxidized enzyme. This species then undergoes minor spectral changes that, based on flavin difference spectra defined in the presence of 3OHKyn, can be correlated with product release. The oxidative half-reaction observed in the presence of saturating benzoylalanine or m-nitrobenzoylalanine also shows the accumulation of a peroxyflavin species that then decays to yield hydrogen peroxide without hydroxylation.

  11. 3-Chloro-1,2-propanediol biodegradation by Ca-alginate immobilized Pseudomonas putida DSM 437 cells applying different processes: mass transfer effects.

    Science.gov (United States)

    Konti, Aikaterini; Mamma, Diomi; Hatzinikolaou, Dimitios G; Kekos, Dimitris

    2016-10-01

    3-Chloro-1,2-propanediol (3-CPD) biodegradation by Ca-alginate immobilized Pseudomonas putida cells was performed in batch system, continuous stirred tank reactor (CSTR), and packed-bed reactor (PBR). Batch system exhibited higher biodegradation rates and 3-CPD uptakes compared to CSTR and PBR. The two continuous systems (CSTR and PBR) when compared at 200 mg/L 3-CPD in the inlet exhibited the same removal of 3-CPD at steady state. External mass-transfer limitations are found negligible at all systems examined, since the observable modulus for external mass transfer Ω ≪ 1 and the Biot number Bi > 1. Intra-particle diffusion resistance had a significant effect on 3-CPD biodegradation in all systems studied, but to a different extent. Thiele modulus was in the range of 2.5 in batch system, but it was increased at 11 when increasing cell loading in the beads, thus lowering significantly the respective effectiveness factor. Comparing the systems at the same cell loading in the beads PBR was less affected by internal diffusional limitations compared to CSTR and batch system, and, as a result, exhibited the highest overall effectiveness factor.

  12. Effectiveness of cleaning and sanitizing procedures in controlling the adherence of Pseudomonas fluorescens, Salmonella Enteritidis, and Staphylococcus aureus to domestic kitchen surfaces Eficiência dos procedimentos de limpeza e de sanitização no controle da adesão de Pseudomonas fluorescens, Salmonella Enteritidis e Staphylococcus aureus em superfícies usadas em cozinhas domésticas

    Directory of Open Access Journals (Sweden)

    Iara Dias Silva

    2010-03-01

    Full Text Available The effectiveness of cleaning and sanitizing procedures in controlling Staphylococcus aureus, Salmonella Enteritidis, and Pseudomonasfluorescens adhered to granite and stainless steel was evaluated. There was no significant difference (p > 0.05 in the adherence of pure cultures of these microorganisms to stainless steel. The numbers of P. fluorescens and S. Enteritidis adhered to granite were greater (p 0.05 between the surfaces. However, a significant difference was observed (p A eficiência dos procedimentos de limpeza e sanitização no controle de Staphylococcus aureus, Salmonella Enteritidis e Pseudomonasfluorescens aderidas em granito e aço inoxidável foi avaliada. Não houve diferença significativa (p > 0,05 na adesão destes microrganismos quando em cultura pura, em aço inoxidável. O número de células aderidas de P. fluorescens e S. Enteritidis foi maior (p 0,05 entre as superfícies. Entretanto, observou-se uma diferença (p < 0,05 entre as soluções sanitizantes utilizadas. Hipoclorito de sódio e ácido peracético apresentaram maior ação bactericida (p < 0,05 que o composto de amônia quaternária. Observou-se que S. aureus apresentou menor resistência à ação desses sanitizantes. Os resultados mostram a importância da adequada realização dos procedimentos de limpeza e sanitização para evitar a adesão bacteriana e formação de biofilme.

  13. Alginate-modifying enzymes: Biological roles and biotechnological uses

    Directory of Open Access Journals (Sweden)

    Helga eErtesvåg

    2015-05-01

    Full Text Available Alginate denotes a group of industrially important 1-4-linked biopolymers composed of the C-5-epimers β-D-mannuronic acid (M and α-L-guluronic acid (G. The polysaccharide is manufactured from brown algae where it constitutes the main structural cell wall polymer. The physical properties of a given alginate molecule, e.g. gel-strength, water-binding capacity, viscosity and biocompatibility, are determined by polymer length, the relative amount and distribution of G residues and the acetyl content, all of which are controlled by alginate modifying enzymes. Alginate has also been isolated from some bacteria belonging to the genera Pseudomonas and Azotobacter, and bacterially synthesized alginate may be O-acetylated at O-2 and/or O-3. Initially, alginate is synthesized as polymannuronic acid, and some M residues are subsequently epimerized to G residues. In bacteria a mannuronan C-5-epimerase (AlgG and an alginate acetylase (AlgX are integral parts of the protein complex necessary for alginate polymerisation and export. All alginate-producing bacteria use periplasmic alginate lyases to remove alginate molecules aberrantly released to the periplasm. Alginate lyases are also produced by organisms that utilize alginate as carbon source. Most alginate-producing organisms encode more than one mannuronan C-5 epimerase, each introducing its specific pattern of G residues. Acetylation protects against further epimerization and from most alginate lyases. One enzyme with alginate deacetylase activity from Pseudomonas syringae has been reported. Functional and structural studies reveal that alginate lyases and epimerases have related enzyme mechanisms and catalytic sites. Alginate lyases are now utilized as tools for alginate characterization. Secreted epimerases have been shown to function well in vitro, and have been engineered further in order to obtain enzymes that can provide alginates with new and desired properties for use in medical and

  14. Alginate oligosaccharides

    DEFF Research Database (Denmark)

    Falkeborg, Mia; Cheong, Ling-Zhi; Gianfico, Carlo

    2014-01-01

    the presence of the conjugated alkene acid structure formed during enzymatic depolymerization. According to the resonance hybrid theory, the parent radicals of AOs are delocalized through allylic rearrangement, and as a consequence, the reactive intermediates are stabilized. AOs were weak ferrous ion chelators......Alginate oligosaccharides (AOs) prepared from alginate, by alginate lyase-mediated depolymerization, were structurally characterized by mass spectrometry, infrared spectrometry and thin layer chromatography. Studies of their antioxidant activities revealed that AOs were able to completely (100....... This work demonstrated that AOs obtained from a facile enzymatic treatment of abundant alginate is an excellent natural antioxidant, which may find applications in the food industry....

  15. Proteinase production in Pseudomonas fluorescens ON2 is affected by carbon sources and allows surface-attached but not planktonic cells to utilize protein for growth in lake water

    DEFF Research Database (Denmark)

    Nicolaisen, Mette Haubjerg; Worm, Jakob; Jørgensen, Niels O. G.

    2012-01-01

    -colonies were able to utilize this resource, while planktonic cells were not. Our experiments are the first to experimentally support models predicting that production of extra-cellular enzymes in dilute environments may be a waste of resources, whereas it represents a favorable feeding strategy in organic...... and there was no evidence for cell density-regulated or starvation-induced proteinase production. Proteinase was produced in the absence of an organic nitrogen source, and citrate had a negative while glucose had a positive effect on the production. Hence P. fluorescens ON2 seems to exploit protein sources by expressing...

  16. Interaction of bacteria-feeding soil flagellates and Pseudomonas spp

    DEFF Research Database (Denmark)

    Pedersen, Annette; Ekelund, Flemming; Johansen, Anders

    2010-01-01

    Pseudomonas strains may be used as alternatives to fungicides as some of them produce secondary metabolites, which can inhibit growth of plant pathogenic fungi. Increased knowledge of non-target effects of the antagonistic bacteria on other soil organisms as well as of the survival and predation...... resistance of the antagonistic bacteria is necessary for risk assessment and increased performance of antagonistic bacteria as biological control agents. In the present study, we aimed to investigate the difference between Pseudomonas spp. with respect to their predation resistance to and effects...... on the three different and common soil flagellates Bodo caudatus, Cercomonas longicauda, and Neocercomonas jutlandica. Two antagonistic Pseudomonas: Pseudomonas fluorescens CHA0 and P. fluorescens DR54 and two positive control strains: P. fluorescens DSM 50090T and Pseudomonas chlororaphis ATCC 43928 were...

  17. Control biológico del marchitamiento vascular causado por Fusarium oxysporum f. sp. phaseoli en fríjol Phaseolus vulgaris L., mediante la acción combinada de Entrophospora colombiana, Trichoderma sp. Y Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    Avendaño Camila

    2006-06-01

    Hidden="false" UnhideWhenUsed="false" QFormat="true" Name="Intense Emphasis" /> Entrophospora colombiana, Trichoderma sp., Pseudomonas fluorescens y una combinación de estos antagonistas fueron evaluados como biocontroladores de Fusarium oxysporumf. sp. Phaseoli en plantas de fríjol de la variedad ‘ICA Tundama’. El ensayo se estableció en una casa de malla del Programa nacional de manejo integrado de plagas (MIP de la Corporación Colombiana de Investigación Agropecuaria (Corpoica, en el Centro de Investigación Tibaitat

  18. Mining Genomes of Biological Control Strains of Pseudomonas spp.: Unexpected Gems and Tailings

    Science.gov (United States)

    The biocontrol bacterium Pseudomonas fluorescens Pf-5 suppresses numerous soilborne plant diseases and produces an array of structurally-characterized secondary metabolites that are toxic to plant pathogenic bacteria, fungi and Oomycetes. Biosynthetic gene clusters for these metabolites compose nea...

  19. Cost modeling of pseudomonoas fluorescens and pseudomonoas chlororphis biocontrol for competitive exclusion of salmonella enterica on tomatoes

    Science.gov (United States)

    Biocontrol measures may enhance postharvest interventions, however; published research on process-based models for biocontrol of foodborne pathogens on produce is limited. The aim of this research was to develop cost model estimates for competitive exclusion process using Pseudomonas fluorescens and...

  20. Optimization of alkaline protease production from Pseudomonas ...

    African Journals Online (AJOL)

    PRECIOUS

    2009-12-15

    Dec 15, 2009 ... protease production was 37°C at pH 9, with 2% inoculum in the medium for 24 h. .... Positive. Catalase test. Positive ... The enzyme activity gradually decreases from ... Effect of temperature on protease production by Pseudomonas fluorescens. 0 .... between RNA polymerase and upstream promotes DNA.

  1. Comparative evaluation of organic formulations of Pseudomonas ...

    African Journals Online (AJOL)

    An experiment was conducted in the laboratory and farm of the Department of Biotechnology, Gauhati University, to explore the potentiality of various organic formulations of Pseudomonas fluorescens (Pf) and to manage bacterial wilt disease of brinjal (Solanum melongena L.) under local conditions. Different organic ...

  2. Heterogeneity of heat-resistant proteases from milk Pseudomonas species.

    Science.gov (United States)

    Marchand, Sophie; Vandriesche, Gonzalez; Coorevits, An; Coudijzer, Katleen; De Jonghe, Valerie; Dewettinck, Koen; De Vos, Paul; Devreese, Bart; Heyndrickx, Marc; De Block, Jan

    2009-07-31

    Pseudomonas fragi, Pseudomonas lundensis and members of the Pseudomonas fluorescens group may spoil Ultra High Temperature (UHT) treated milk and dairy products, due to the production of heat-stable proteases in the cold chain of raw milk. Since the aprX gene codes for a heat-resistant protease in P. fluorescens, the presence of this gene has also been investigated in other members of the genus. For this purpose an aprX-screening PCR test has been developed. Twenty-nine representatives of important milk Pseudomonas species and thirty-five reference strains were screened. In 42 out of 55 investigated Pseudomonas strains, the aprX gene was detected, which proves the potential of the aprX-PCR test as a screening tool for potentially proteolytic Pseudomonas strains in milk samples. An extensive study of the obtained aprX-sequences on the DNA and the amino acid level, however, revealed a large heterogeneity within the investigated milk isolates. Although this heterogeneity sets limitations to a general detection method for all proteolytic Pseudomonas strains in milk, it offers a great potential for the development of a multiplex PCR screening test targeting individual aprX-genes. Furthermore, our data illustrated the potential use of the aprX gene as a taxonomic marker, which may help in resolving the current taxonomic deadlock in the P. fluorescens group.

  3. Novel Pseudomonas fluorescens Septic Sacroiliitis in a Healthy Soldier

    Science.gov (United States)

    2013-08-01

    acute pathology or sacroiliac (SI) joint involvement, and a colonoscopy with random rectal mucosal biopsy was normal. Two months later, he experienced...an acute episode of excru- ciating, focal left sacroiliac pain after a fall from a seated height of two feet, necessitating this hospital admission...bone marrow edema of the left sacroiliac joint, and muscle edema of the left iliacus and gluteus medius. FIGURE 2. CT-guided arthrocentesis of the left

  4. Pseudomonas fluorescens strains selectively suppress annual bluegrass (Poa annua L.)

    Science.gov (United States)

    Annual bluegrass (Poa annua L.) is a cool-season annual grass that is a major weed species in turf, turfgrass-seed production, sod production, and golf courses of the western United States. There are few selective herbicides available for the management of annual bluegrass. While the life cycles o...

  5. Interactions between biosurfactant-producing Pseudomonas and Phytophthora species

    OpenAIRE

    Tran, H.

    2007-01-01

    Fluorescent Pseudomonas bacteria produce a wide variety of antimicrobial metabolites, including soap-like compounds referred to as biosurfactants. The results of this thesis showed that biosurfactant-producing Pseudomonas bacteria are effective in controlling Phytophthora foot rot disease of black pepper in Vietnam and promote root and shoot development of the ‘King of Spices’. Biosurfactant-producing P. fluorescens strain SS101 was also effective in controlling tomato late blight caused by P...

  6. Disruption of transporters affiliated with enantio-pyochelin biosynthesis gene cluster of Pseudomonas protegens Pf-5 has pleiotropic effects

    Science.gov (United States)

    Pseudomonas protegens Pf-5 (formerly Pseudomonas fluorescens) is a biocontrol bacterium that produces the siderophore enantio-pyochelin under conditions of iron starvation in a process that is often accompanied by the secretion of its biosynthesis intermediates, salicylic acid and dihydroaeruginoic ...

  7. Capsule production by Pseudomonas aeruginosa

    Energy Technology Data Exchange (ETDEWEB)

    Lynn, A.R.

    1984-01-01

    Mucoid strains of Pseudomonas aeruginosa, associated almost exclusively with chronic respiratory infections in patients with cystic fibrosis, possess a capsule composed of alginic acid similar to one produced by Azotobacter vinelandii. Recent reports have provided evidence that the biosynthetic pathway for alginate in P. aeruginosa may differ from the pathway proposed for A. vinelandii in that synthesis in P. aeruginosa may occur by way of the Entner-Doudoroff pathway. Incorporation of isotope from (6-/sup 14/C)glucose into alginate by both P. aueroginosa and A. vinelandii was 10-fold greater than that from either (1-/sup 14/C)/sup -/ or (2-/sup 14/C)glucose, indicating preferential utilization of the bottom half of the glucose molecule for alginate biosynthesis. These data strongly suggest that the Entner-Doudoroff pathway plays a major role in alginate synthesis in both P. aeruginosa and A. vinelandii. The enzymes of carbohydrate metabolism in mucoid strains of P. aeruginosa appear to be unchanged whether alignate is actively produced or not and activities do not differ significantly from nonmucoid strain PAO.

  8. Characterization of AlgMsp, an alginate lyase from Microbulbifer sp. 6532A.

    Directory of Open Access Journals (Sweden)

    Steven M Swift

    Full Text Available Alginate is a polysaccharide produced by certain seaweeds and bacteria that consists of mannuronic acid and guluronic acid residues. Seaweed alginate is used in food and industrial chemical processes, while the biosynthesis of bacterial alginate is associated with pathogenic Pseudomonas aeruginosa. Alginate lyases cleave this polysaccharide into short oligo-uronates and thus have the potential to be utilized for both industrial and medicinal applications. An alginate lyase gene, algMsp, from Microbulbifer sp. 6532A, was synthesized as an E.coli codon-optimized clone. The resulting 37 kDa recombinant protein, AlgMsp, was expressed, purified and characterized. The alginate lyase displayed highest activity at pH 8 and 0.2 M NaCl. Activity of the alginate lyase was greatest at 50°C; however the enzyme was not stable over time when incubated at 50°C. The alginate lyase was still highly active at 25°C and displayed little or no loss of activity after 24 hours at 25°C. The activity of AlgMsp was not dependent on the presence of divalent cations. Comparing activity of the lyase against polymannuronic acid and polyguluronic acid substrates showed a higher turnover rate for polymannuronic acid. However, AlgMSP exhibited greater catalytic efficiency with the polyguluronic acid substrate. Prolonged AlgMsp-mediated degradation of alginate produced dimer, trimer, tetramer, and pentamer oligo-uronates.

  9. Preparation methods of alginate nanoparticles

    NARCIS (Netherlands)

    Paques, J.P.; Linden, van der E.; Rijn, van C.J.M.; Sagis, L.M.C.

    2014-01-01

    This article reviews available methods for the formation of alginate nano-aggregates, nanocapsules and nanospheres. Primarily, alginate nanoparticles are being prepared by two methods. In the “complexation method”, complex formation on the interface of an oil droplet is used to form alginate

  10. Antibiotic and biosurfactant properties of cyclic lipopeptides produced by fluorescent Pseudomonas spp. from the sugar beet rhizosphere

    DEFF Research Database (Denmark)

    Nielsen, T H; Sørensen, D; Tobiasen, C

    2002-01-01

    Cyclic lipopeptides (CLPs) with antibiotic and biosurfactant properties are produced by a number of soil bacteria, including fluorescent Pseudomonas spp. To provide new and efficient strains for the biological control of root-pathogenic fungi in agricultural crops, we isolated approximately 600...... in the peptide moiety. Production of specific CLPs could be affiliated with Pseudomonas fluorescens strain groups belonging to biotype I, V, or VI. In vitro analysis using both purified CLPs and whole-cell P. fluorescens preparations demonstrated that all CLPs exhibited strong biosurfactant properties...

  11. Chemical sanitizers to control biofilms formed by two Pseudomonas species on stainless steel surface

    Directory of Open Access Journals (Sweden)

    Danila Soares Caixeta

    2012-03-01

    Full Text Available The biofilm formation of Pseudomonas aeruginosa and Pseudomonas fluorescens on AISI 304 stainless steel in the presence of reconstituted skim milk under different temperatures was conducted, and the potential of three chemical sanitizers in removing the mono-species biofilms formed was compared. Pseudomonas aeruginosa cultivated in skim milk at 28 °C presented better growth rate (10.4 log CFU.mL-1 when compared with 3.7 and 4.2 log CFU.mL-1 for P. aeruginosa and P. fluorescens cultivated at 7 °C, respectively. Pseudomonas aeruginosa formed biofilm when cultivated at 28 °C. However, only the adhesion of P. aeruginosa and P. fluorescens was observed when incubated at 7 °C. The sodium dichloroisocyanurate was the most efficient sanitizer in the reduction of the adhered P. aeruginosa cells at 7 and 28 °C and those on the biofilm, respectively. The hydrogen peroxide was more effective in the reduction of adhered cells of P. fluorescens at 7 °C.

  12. Catalytic Mechanism and Mode of Action of the Periplasmic Alginate Epimerase AlgG

    NARCIS (Netherlands)

    Wolfram, Francis; Kitova, Elena N.; Robinson, Howard; Walvoort, Marthe T. C.; Codee, Jeroen D. C.; Klassen, John S.; Howell, P. Lynne

    2014-01-01

    Background: The alginate epimerase AlgG converts mannuronate to its C5 epimer guluronate at the polymer level. Results: The structure of Pseudomonas syringae AlgG has been determined, and the protein has been functionally characterized. Conclusion: His(319) acts as the catalytic base, whereas

  13. Chronic Pseudomonas aeruginosa lung infection in normal and athymic rats

    DEFF Research Database (Denmark)

    Johansen, H K; Espersen, F; Pedersen, S S

    1993-01-01

    We have compared a chronic lung infection with Pseudomonas aeruginosa embedded in alginate beads in normal and athymic rats with an acute infection with free live P. aeruginosa bacteria. The following parameters were observed and described: mortality, macroscopic and microscopic pathologic changes...

  14. Matrix evaluation for Pseudomonas spp. immobilisation in phenol bioremediation

    Directory of Open Access Journals (Sweden)

    Leonel Chitiva Urbina

    2003-07-01

    Full Text Available Pseudomonas spp. were cultivated in a free cell suspension and also immobilised in three different matrices to observe the influence of a contaminant like phenol on degradation velocity and compare each one's results. Polyurethane polymers, alginate (Manohar et al, 2001 and a mixture of alginate and polyvinyl alcohol (Doria et al, 2002 were selected and tested as matrices; all of them proved viable as matrices for cell immobilisation. Pseudomonas were cultivated in an initial 10 cfu/ml concentration in each one of the matrices for comparison purposes and in a medium without matrix; all mediums were supplemented with a minimum salt medium and 200 ppm phenol. A removal time of 23 days was observed in the medium without matrix, 15 days in the polyurethane matrix and 7 days in the alginate matrices. Improved removal times were observed in all matrices when compared to the free cell suspension.

  15. Alginate Production from Alternative Carbon Sources and Use of Polymer Based Adsorbent in Heavy Metal Removal

    Directory of Open Access Journals (Sweden)

    Çiğdem Kıvılcımdan Moral

    2016-01-01

    Full Text Available Alginate is a biopolymer composed of mannuronic and guluronic acids. It is harvested from marine brown algae; however, alginate can also be synthesized by some bacterial species, namely, Azotobacter and Pseudomonas. Use of pure carbohydrate sources for bacterial alginate production increases its cost and limits the chance of the polymer in the industrial market. In order to reduce the cost of bacterial alginate production, molasses, maltose, and starch were utilized as alternative low cost carbon sources in this study. Results were promising in the case of molasses with the maximum 4.67 g/L of alginate production. Alginates were rich in mannuronic acid during early fermentation independent of the carbon sources while the highest guluronic acid content was obtained as 68% in the case of maltose. The polymer was then combined with clinoptilolite, which is a natural zeolite, to remove copper from a synthetic wastewater. Alginate-clinoptilolite beads were efficiently adsorbed copper up to 131.6 mg Cu2+/g adsorbent at pH 4.5 according to the Langmuir isotherm model.

  16. Diversity of Pseudomonas Genomes, Including Populus-Associated Isolates, as Revealed by Comparative Genome Analysis.

    Science.gov (United States)

    Jun, Se-Ran; Wassenaar, Trudy M; Nookaew, Intawat; Hauser, Loren; Wanchai, Visanu; Land, Miriam; Timm, Collin M; Lu, Tse-Yuan S; Schadt, Christopher W; Doktycz, Mitchel J; Pelletier, Dale A; Ussery, David W

    2016-01-01

    The Pseudomonas genus contains a metabolically versatile group of organisms that are known to occupy numerous ecological niches, including the rhizosphere and endosphere of many plants. Their diversity influences the phylogenetic diversity and heterogeneity of these communities. On the basis of average amino acid identity, comparative genome analysis of >1,000 Pseudomonas genomes, including 21 Pseudomonas strains isolated from the roots of native Populus deltoides (eastern cottonwood) trees resulted in consistent and robust genomic clusters with phylogenetic homogeneity. All Pseudomonas aeruginosa genomes clustered together, and these were clearly distinct from other Pseudomonas species groups on the basis of pangenome and core genome analyses. In contrast, the genomes of Pseudomonas fluorescens were organized into 20 distinct genomic clusters, representing enormous diversity and heterogeneity. Most of our 21 Populus-associated isolates formed three distinct subgroups within the major P. fluorescens group, supported by pathway profile analysis, while two isolates were more closely related to Pseudomonas chlororaphis and Pseudomonas putida. Genes specific to Populus-associated subgroups were identified. Genes specific to subgroup 1 include several sensory systems that act in two-component signal transduction, a TonB-dependent receptor, and a phosphorelay sensor. Genes specific to subgroup 2 contain hypothetical genes, and genes specific to subgroup 3 were annotated with hydrolase activity. This study justifies the need to sequence multiple isolates, especially from P. fluorescens, which displays the most genetic variation, in order to study functional capabilities from a pangenomic perspective. This information will prove useful when choosing Pseudomonas strains for use to promote growth and increase disease resistance in plants. Copyright © 2015 Jun et al.

  17. Kefiran-alginate gel microspheres for oral delivery of ciprofloxacin.

    Science.gov (United States)

    Blandón, Lina M; Islan, German A; Castro, Guillermo R; Noseda, Miguel D; Thomaz-Soccol, Vanete; Soccol, Carlos R

    2016-09-01

    Ciprofloxacin is a broad-spectrum antibiotic associated with gastric and intestinal side effects after extended oral administration. Alginate is a biopolymer commonly employed in gel synthesis by ionotropic gelation, but unstable in the presence of biological metal-chelating compounds and/or under dried conditions. Kefiran is a microbial biopolymer able to form gels with the advantage of displaying antimicrobial activity. In the present study, kefiran-alginate gel microspheres were developed to encapsulate ciprofloxacin for antimicrobial controlled release and enhanced bactericidal effect against common pathogens. Scanning electron microscopy (SEM) analysis of the hybrid gel microspheres showed a spherical structure with a smoother surface compared to alginate gel matrices. In vitro release of ciprofloxacin from kefiran-alginate microspheres was less than 3.0% and 5.0% at pH 1.2 (stomach), and 5.0% and 25.0% at pH 7.4 (intestine) in 3 and 21h, respectively. Fourier transform infrared spectroscopy (FTIR) of ciprofloxacin-kefiran showed the displacement of typical bands of ciprofloxacin and kefiran, suggesting a cooperative interaction by hydrogen bridges between both molecules. Additionally, the thermal analysis of ciprofloxacin-kefiran showed a protective effect of the biopolymer against ciprofloxacin degradation at high temperatures. Finally, antimicrobial assays of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhymurium, and Staphylococcus aureus demonstrated the synergic effect between ciprofloxacin and kefiran against the tested microorganisms. Copyright © 2016 Elsevier B.V. All rights reserved.

  18. Structural basis for alginate secretion across the bacterial outer membrane

    Energy Technology Data Exchange (ETDEWEB)

    Whitney, J.C.; Robinson, H.; Hay, I. D.; Li, C.; Eckford, P. D. W.; Amaya, M. F.; Wood, L. F.; Ohman, D. E.; Bear, C. E.; Rehm, B. H.; Howell, P. L.

    2011-08-09

    Pseudomonas aeruginosa is the predominant pathogen associated with chronic lung infection among cystic fibrosis patients. During colonization of the lung, P. aeruginosa converts to a mucoid phenotype characterized by the overproduction of the exopolysaccharide alginate. Secretion of newly synthesized alginate across the outer membrane is believed to occur through the outer membrane protein AlgE. Here we report the 2.3 {angstrom} crystal structure of AlgE, which reveals a monomeric 18-stranded {beta}-barrel characterized by a highly electropositive pore constriction formed by an arginine-rich conduit that likely acts as a selectivity filter for the negatively charged alginate polymer. Interestingly, the pore constriction is occluded on either side by extracellular loop L2 and an unusually long periplasmic loop, T8. In halide efflux assays, deletion of loop T8 ({Delta}T8-AlgE) resulted in a threefold increase in anion flux compared to the wild-type or {Delta}L2-AlgE supporting the idea that AlgE forms a transport pathway through the membrane and suggesting that transport is regulated by T8. This model is further supported by in vivo experiments showing that complementation of an algE deletion mutant with {Delta}T8-AlgE impairs alginate production. Taken together, these studies support a mechanism for exopolysaccharide export across the outer membrane that is distinct from the Wza-mediated translocation observed in canonical capsular polysaccharide export systems.

  19. Structural Basis for Alginate Secretion Across the Bacterial Outer Membrane

    Energy Technology Data Exchange (ETDEWEB)

    J Whitney; I Hay; C Li; P Eckford; H Robinson; M Amaya; L Wood; D Ohman; C Bear; et al.

    2011-12-31

    Pseudomonas aeruginosa is the predominant pathogen associated with chronic lung infection among cystic fibrosis patients. During colonization of the lung, P. aeruginosa converts to a mucoid phenotype characterized by the overproduction of the exopolysaccharide alginate. Secretion of newly synthesized alginate across the outer membrane is believed to occur through the outer membrane protein AlgE. Here we report the 2.3 {angstrom} crystal structure of AlgE, which reveals a monomeric 18-stranded {beta}-barrel characterized by a highly electropositive pore constriction formed by an arginine-rich conduit that likely acts as a selectivity filter for the negatively charged alginate polymer. Interestingly, the pore constriction is occluded on either side by extracellular loop L2 and an unusually long periplasmic loop, T8. In halide efflux assays, deletion of loop T8 ({Delta}T8-AlgE) resulted in a threefold increase in anion flux compared to the wild-type or {Delta}L2-AlgE supporting the idea that AlgE forms a transport pathway through the membrane and suggesting that transport is regulated by T8. This model is further supported by in vivo experiments showing that complementation of an algE deletion mutant with {Delta}T8-AlgE impairs alginate production. Taken together, these studies support a mechanism for exopolysaccharide export across the outer membrane that is distinct from the Wza-mediated translocation observed in canonical capsular polysaccharide export systems.

  20. Differentiation of isolated native of desulphurizators Pseudomonas by means of the study of the profile of fatty acids

    International Nuclear Information System (INIS)

    Silva Gomez, Edelberto; Morales P, Alicia Lucia; Callejas Castro Solange; Florez Sandoval Maria Cecilia; Rodriguez Wilson

    2000-01-01

    The content of cellular fatty acids was determined by HRGC of twelve Colombian isolated Pseudomonas aeruginosa 17,18, 19, 20, 21, 22 and 103 pseudomonas sp 23,24,25,26 and 27 with desulphurization capacity, pseudomonas aeruginosa ATCC 9027 and 10145, pseudomonas sp ATCC 39327 and pseudomonas fluorescens. Fifty-three different types of fatty acids were found, among saturated and unsaturated of lineal chain, and mainly hydroxy acids and ramified. Of these, 17 have not been described in the literature for this genus. A group of 6 acids was presented with more frequency (15:0; 16:0; 16:1; 17:1; 3-OH 16:0 and 2-OH 15:0) in more than 75% of the pseudomonas studied. The studied microorganisms are related because they share the presence of some characteristic fatty acids, that which allows assuring that they belong to same taxonomic unit. The analysis cluster developed by means of plotting in dendrograms of the qualitative and quantitative contents of the acids fatty totals showed the formation of two attaches, the I conformed by ATCC 39327 and the isolated 17 and 25, and the II by Ps. Fluorescens and the isolated one 27. The dendrogram of the hydroxy acids shows the formation of four attaches, the I attache, (isolated 17 and 20), the II A (isolated 22 and 103), the II B (isolated 27 and ps. fluorescens) and the attache III (isolated 18 and 19). In that of the ramified fatty acids the formation of a main attache is observed conformed by four sub-groups, the IA (isolated 17), the I B (isolated 18 and 24), the I C (isolated 19 and 25 and Ps. fluorescens) and I D (isolated 27). These results show that the isolated 27,25,24, 19, 18 and 17, in their order, have narrow relationship to Ps. fluorescens

  1. Rhizosphere competence of wild-type and genetically-engineered Pseudomonas brassicacearum is affected by the crop species

    Science.gov (United States)

    2,4-diacetylphloroglucinol (2,4-DAPG)-producing Pseudomonas brassicacearum Q8r1-96 is a highly effective biocontrol agent of take-all disease of wheat. Strain Z30-97, a recombinant derivative of Q8r1-96 containing the phzABCDEFG operon from P. synxantha (formerly P. fluorescens) 2-79 inserted into ...

  2. Effect of Genetically Modified Pseudomonas putida WCS358r on the Fungal Rhizosphere Microflora of Field-Grown Wheat

    NARCIS (Netherlands)

    Glandorf, D.C.M.; Verheggen, Patrick; Jansen, Timo; Jorritsma, J.-W.; Smit, Eric; Leeflang, Paula; Wernars, Karel; Thomashow, L.S.; Laureijs, Eric; Thomas-Oates, J.E.; Bakker, P.A.H.M.; Loon, L.C. van

    2001-01-01

    We released genetically modified Pseudomonas putida WCS358r into the rhizospheres of wheat plants. The two genetically modified derivatives, genetically modified microorganism (GMM) 2 and GMM 8, carried the phz biosynthetic gene locus of strain P. fluorescens 2-79 and constitutively produced the

  3. Pseudomonas - Fact Sheet

    OpenAIRE

    Public Health Agency

    2012-01-01

    Fact sheet on Pseudomonas, including:What is Pseudomonas?What infections does it cause?Who is susceptible to pseudomonas infection?How will I know if I have pseudomonas infection?How can Pseudomonas be prevented from spreading?How can I protect myself from Pseudomonas?How is Pseudomonas infection treated?

  4. Chemical sanitizers to control biofilms formed by two Pseudomonas species on stainless steel surface Sanificantes químicos no controle de biofilmes formados por duas espécies de Pseudomonas em superfície de aço inoxidável

    Directory of Open Access Journals (Sweden)

    Danila Soares Caixeta

    2012-03-01

    Full Text Available The biofilm formation of Pseudomonas aeruginosa and Pseudomonas fluorescens on AISI 304 stainless steel in the presence of reconstituted skim milk under different temperatures was conducted, and the potential of three chemical sanitizers in removing the mono-species biofilms formed was compared. Pseudomonas aeruginosa cultivated in skim milk at 28 °C presented better growth rate (10.4 log CFU.mL-1 when compared with 3.7 and 4.2 log CFU.mL-1 for P. aeruginosa and P. fluorescens cultivated at 7 °C, respectively. Pseudomonas aeruginosa formed biofilm when cultivated at 28 °C. However, only the adhesion of P. aeruginosa and P. fluorescens was observed when incubated at 7 °C. The sodium dichloroisocyanurate was the most efficient sanitizer in the reduction of the adhered P. aeruginosa cells at 7 and 28 °C and those on the biofilm, respectively. The hydrogen peroxide was more effective in the reduction of adhered cells of P. fluorescens at 7 °C.A capacidade de adesão e formação de biofilme por Pseudomonas aeruginosa e Pseudomonas fluorescens em aço inoxidável AISI 304, na presença de leite desnatado resconstituído sobre diferentes temperaturas foi conduzido e o potencial de três sanificantes químicos na remoção de biofilmes monoespécies foi comparado. Pseudomonas aeruginosa cultivada em leite desnatado a 28 °C apresentou melhor crescimento (10,4 log UFC.mL-1 quando comparado com 3,7 and 4,2 log UFC.mL-1 para P. aeruginosa e P. fluorescens cultivadas a 7 °C, respectivamente. Pseudomonas aeruginosa formou biofilme quando cultivada a 28 °C. Contudo foi observado somente adesão de P. aeruginosa e P. fluorescens quando incubada a 7 °C. O dicloroisocianurato de sódio foi o sanificante mais eficiente na redução de células aderidas e em biofilme de P. aeruginosa a 7 e 28 °C, respectivamente. O peróxido de hidrogênio foi o mais eficiente na redução de células aderidas de P. fluorescens a 7 °C.

  5. Switching catalysis from hydrolysis to perhydrolysis in P. fluorescens esterase

    Science.gov (United States)

    Yin, De Lu (Tyler); Bernhardt, Peter; Morley, Krista L.; Jiang, Yun; Cheeseman, Jeremy D.; Purpero, Vincent; Schrag, Joseph D.; Kazlauskas, Romas J.

    2010-01-01

    Many serine hydrolases catalyze perhydrolysis – the reversible formation of per-acids from carboxylic acids and hydrogen peroxide. Recently we showed that a single amino acid substitution in the alcohol binding pocket - L29P - in Pseudomonas fluorescens (SIK WI) aryl esterase (PFE) increased the specificity constant of PFE for peracetic acid formation >100-fold [Bernhardt et al. Angew. Chem. Intl. Ed. 2005, 44, 2742]. In this paper, we extend this work to address the three following questions. First, what is the molecular basis of the increase in perhydrolysis activity? We previously proposed that the L29P substitution creates a hydrogen bond between the enzyme and hydrogen peroxide in the transition state. Here we report two x-ray structures of L29P PFE that support this proposal. Both structures show a main chain carbonyl oxygen closer to the active-site serine as expected. One structure further shows acetate in the active site in an orientation consistent with reaction by an acyl-enzyme mechanism. We also detected an acyl-enzyme intermediate in the hydrolysis of ε-caprolactone by mass spectrometry. Second, can we further increase perhydrolysis activity? We discovered that the reverse reaction – hydrolysis of peracetic acid to acetic acid and hydrogen peroxide – occurs at nearly the diffusion limited rate. Since the reverse reaction cannot increase further, neither can the forward reaction. Consistent with this prediction, two variants with additional amino acid substitutions showed two fold higher kcat, but Km also increased so the specificity constant, kcat/Km, remained similar. Third, how does the L29P substitution change the esterase activity? Ester hydrolysis decreased for most esters (75-fold for ethyl acetate), but not for methyl esters. In contrast, L29P PFE catalyzed hydrolysis of ε-caprolactone five times more efficiently than wild-type PFE. Molecular modeling suggests that moving the carbonyl group closer to the active site blocks access for

  6. Response to gaseous NO2 air pollutant of P. fluorescens airborne strain MFAF76a and clinical strain MFN1032

    Directory of Open Access Journals (Sweden)

    Tatiana eKondakova

    2016-03-01

    Full Text Available Human exposure to nitrogen dioxide (NO2, an air pollutant of increasing interest in biology, results in several toxic effects to human health and also to the air microbiota. The aim of this study was to investigate the bacterial response to gaseous NO2. Two Pseudomonas fluorescens strains, namely the airborne strain MFAF76a and the clinical strain MFN1032 were exposed to 0.1, 5 or 45 ppm concentrations of NO2, and their effects on bacteria were evaluated in terms of motility, biofilm formation, antibiotic resistance, as well as expression of several chosen target genes. While 0.1 and 5 ppm of NO2 did not lead to any detectable modification in the studied phenotypes of the two bacteria, several alterations were observed when the bacteria were exposed to 45 ppm of gaseous NO2. We thus chose to focus on this high concentration. NO2-exposed P. fluorescens strains showed reduced swimming motility, and decreased swarming in case of the strain MFN1032. Biofilm formed by NO2-treated airborne strain MFAF76a showed increased maximum thickness compared to non-treated cells, while NO2 had no apparent effect on the clinical MFN1032 biofilm structure. It is well known that biofilm and motility are inversely regulated by intracellular c-di-GMP level. The c-di-GMP level was however not affected in response to NO2 treatment. Finally, NO2-exposed P. fluorescens strains were found to be more resistant to ciprofloxacin and chloramphenicol. Accordingly, the resistance nodulation cell division (RND MexEF-OprN efflux pump encoding genes were highly upregulated in the two P. fluorescens strains. Noticeably, similar phenotypes had been previously observed following a NO treatment. Interestingly, an hmp-homologue gene in P. fluorescens strains MFAF76a and MFN1032 encodes a NO dioxygenase that is involved in NO detoxification into nitrites. Its expression was upregulated in response to NO2, suggesting a possible common pathway between NO and NO2 detoxification. Taken

  7. Physiology and biochemistry of polysaccharide production by Azotobacter vinelandii and Pseudomonas aeruginosa

    Energy Technology Data Exchange (ETDEWEB)

    Annison, G.

    1985-01-01

    The extracellular production and the composition of the acetylated alginates of Azotobacter vinelandii and Pseudomonas aeruginosa were investigated. A reverse-phase HPLC method to determine D-mannuronate/L-guluronate (M/G) ratios in alginates was developed. Comparison between the HPLC procedure and a /sup 1/H-NMR method was made. Growth and alginate production by A. vinelandii in batch and continuous culture were studied. Polysaccharide production was favored in batch culture by reduced aeration rates by by intermediate aeration rates in continuous culture. The partitioning of Ca/sup 45/ in solution and associated with alginates during the growth cycle of A. vinelandii was studied. The change in the composition of polysaccharide produced during growth of A. vinelandii was related to the level of free Ca/sup + +/ which fell rapidly during initial stages of incubation. The degree of acetylation of the polyuronate was shown to be variable. Production of alginates with high O-acetate contents was observed in cultures maintained at high growth rates and high Ca/sup + +/ levels. The degree of acetylation of the polyuronate was not related to the level of epimerization in vivo although in vitro studies demonstrated that chemical acetylation of the alginate inhibited epimerization. Alginate production by clinical isolates of Pseudomonas aeruginosa was shown to be unstable and no polyguluronate was isolated from cultures. It was also noted that Ca/sup + +/ played a less important role in the composition of the polysaccharide.

  8. Antibiotic and biosurfactant properties of cyclic lipopeptides produced by fluorescent Pseudomonas spp. from the sugar beet rhizosphere

    DEFF Research Database (Denmark)

    Nielsen, T.H.; Sørensen, D.; Tobiasen, C.

    2002-01-01

    Cyclic lipopeptides (CLPs) with antibiotic and biosurfactant properties are produced by a number of soil bacteria, including fluorescent Pseudomonas spp. To provide new and efficient strains for the biological control of root-pathogenic fungi in agricultural crops, we isolated approximately 600...... fluorescent Pseudomonas spp. from two different agricultural soils by using three different growth media. CLP production was observed in a large proportion of the strains (approximately 60%) inhabiting the sandy soil, compared to a low proportion (approximately 6%) in the loamy soil. Chemical structure...... in the peptide moiety. Production of specific CLPs could be affiliated with Pseudomonas fluorescens strain groups belonging to biotype I, V, or VI. In vitro analysis using both purified CLPs and whole-cell P. fluorescens preparations demonstrated that all CLPs exhibited strong biosurfactant properties...

  9. Chromosomal insertion of the entire Escherichia coli lactose operon, into two strains of Pseudomonas, using a modified mini-Tn5 delivery system

    DEFF Research Database (Denmark)

    Hansen, L. H.; Sørensen, S. J.; Jensen, Lars Bogø

    1997-01-01

    A 12-kb PstI fragment including the entire E. coli lactose operon (lacIPOZYA) was inserted in one copy into the chromosome of Pseudomonas putida, Pseudomonas fluorescens and an E. coli strain with lac(-) phenotype. This was made possible by improvements of an already existing mini-Tn5 transposon...... flanked by NotI sites needed in the mini-Tn5 delivery system; (b) the generation of E. coli nonlysogenic strains expressing the pi protein thus being capable of maintaining and delivering R6K-based mini-Tn5 vectors to other E. coli strains; (c) the successful insertion of the E. coli lactose operon...... into the P. fluorescens chromosome giving P. fluorescens the ability to grow on lactose; (d) evidence from Southern blotting that contradicts the assumption that the mini-Tn5 delivery system always creates one-copy inserts. These improvements allow insertion of large DNA fragments encoding highly expressed...

  10. Pseudomonas aeruginosa biofilms in cystic fibrosis

    DEFF Research Database (Denmark)

    Høiby, Niels; Ciofu, Oana; Bjarnsholt, Thomas

    2010-01-01

    The persistence of chronic Pseudomonas aeruginosa lung infections in cystic fibrosis (CF) patients is due to biofilm-growing mucoid (alginate-producing) strains. A biofilm is a structured consortium of bacteria, embedded in a self-produced polymer matrix consisting of polysaccharide, protein...... and DNA. In CF lungs, the polysaccharide alginate is the major part of the P. aeruginosa biofilm matrix. Bacterial biofilms cause chronic infections because they show increased tolerance to antibiotics and resist phagocytosis, as well as other components of the innate and the adaptive immune system....... As a consequence, a pronounced antibody response develops, leading to immune complex-mediated chronic inflammation, dominated by polymorphonuclear leukocytes. The chronic inflammation is the major cause of the lung tissue damage in CF. Biofilm growth in CF lungs is associated with an increased frequency...

  11. Variation of the Pseudomonas community structure on oak leaf lettuce during storage detected by culture-dependent and -independent methods.

    Science.gov (United States)

    Nübling, Simone; Schmidt, Herbert; Weiss, Agnes

    2016-01-04

    The genus Pseudomonas plays an important role in the lettuce leaf microbiota and certain species can induce spoilage. The aim of this study was to investigate the occurrence and diversity of Pseudomonas spp. on oak leaf lettuce and to follow their community shift during a six day cold storage with culture-dependent and culture-independent methods. In total, 21 analysed partial Pseudomonas 16S rRNA gene sequences matched closely (> 98.3%) to the different reference strain sequences, which were distributed among 13 different phylogenetic groups or subgroups within the genus Pseudomonas. It could be shown that all detected Pseudomonas species belonged to the P. fluorescens lineage. In the culture-dependent analysis, 73% of the isolates at day 0 and 79% of the isolates at day 6 belonged to the P. fluorescens subgroup. The second most frequent group, with 12% of the isolates, was the P. koreensis subgroup. This subgroup was only detected at day 0. In the culture-independent analysis the P. fluorescens subgroup and P. extremaustralis could not be differentiated by RFLP. Both groups were most abundant and amounted to approximately 46% at day 0 and 79% at day 6. The phytopathogenic species P. salmonii, P. viridiflava and P. marginalis increased during storage. Both approaches identified the P. fluorescens group as the main phylogenetic group. The results of the present study suggest that pseudomonads found by plating methods indeed represent the most abundant part of the Pseudomonas community on oak leaf lettuce. Copyright © 2015 Elsevier B.V. All rights reserved.

  12. Disinfectant Efficacy of 0.525% Sodium Hypochlorite and Epimax on Alginate Impression Material.

    Science.gov (United States)

    Choudhury, Gopal Krishna; Chitumalla, Rajkiran; Manual, Litto; Rajalbandi, Santosh Kumar; Chauhan, Mahinder Singh; Talukdar, Pratim

    2018-01-01

    Species of Streptococcus, Escherichia coli, Staphylococcus, Actinomyces, Pseudomonas, Klebsiella, and Candida are commonly seen in the oral cavity. Impression materials are commonly contaminated with microorganisms. The present study was conducted to assess the disinfection efficacy of Epimax and 0.525% sodium hypochlorite on alginate impression over a period of 10 minutes. This study was conducted in the Department of Prosthodontics in the year 2015. An alginate impression material was prepared. For each bacteria species, 15 samples were used. Out of 15 samples, 3 were used by 0.525% sodium hypochlorite for disinfection for 5 minutes and 3 others for 10 minutes. Similarly, 3 samples were used by Epimax for 5 minutes and other 3 for 10 minutes. Three samples were used as controls. Each sample was polluted with Candida albicans, Pseudomonas aeruginosa, and Staphylococcus aureus strains. There was no statistical difference in P. aeruginosa and C. albicans after 5 minutes, whereas S. aureus showed significant difference (p alginate impression material against C. albicans, P. aeruginosa, and S. aureus strains. However, Epimax was found to be more effective against S. aureus as compared with 0.525% sodium hypochlorite. Efficacy of disinfection of sodium hypo-chlorite and Epimax on alginate impression.

  13. Bioremediation of Petroleum hydrocarbon by using Pseudomonas species isolated from Petroleum contaminated soil

    OpenAIRE

    Vijay Kumar; Simranjeet Singh; Anu Manhas; Joginder Singh; Sourav Singla; Parvinder Kaur; Shivika Data; Pritika Negi; Arjun Kalia

    2014-01-01

    A newly isolated strain Pseudomonas fluorescens (Accession number KF 279042.1) have potential in diesel degradation and can be recommended for bioremediation of sites that are contaminated with diesel. This bacterium was characterized on the basis of microbiological, biochemical and molecular analysis. Bacterial growth optimization was studied based on carbon source, nitrogen source, pH and temperature. The strain was selected based on its ability to show growth in medium containing diesel. I...

  14. 21 CFR 582.7187 - Calcium alginate.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 6 2010-04-01 2010-04-01 false Calcium alginate. 582.7187 Section 582.7187 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) ANIMAL... Calcium alginate. (a) Product. Calcium alginate. (b) Conditions of use. This substance is generally...

  15. Efficient functionalization of alginate biomaterials.

    Science.gov (United States)

    Dalheim, Marianne Ø; Vanacker, Julie; Najmi, Maryam A; Aachmann, Finn L; Strand, Berit L; Christensen, Bjørn E

    2016-02-01

    Peptide coupled alginates obtained by chemical functionalization of alginates are commonly used as scaffold materials for cells in regenerative medicine and tissue engineering. We here present an alternative to the commonly used carbodiimide chemistry, using partial periodate oxidation followed by reductive amination. High and precise degrees of substitution were obtained with high reproducibility, and without formation of by-products. A protocol was established using l-Tyrosine methyl ester as a model compound and the non-toxic pic-BH3 as the reducing agent. DOSY was used to indirectly verify covalent binding and the structure of the product was further elucidated using NMR spectroscopy. The coupling efficiency was to some extent dependent on alginate composition, being most efficient on mannuronan. Three different bioactive peptide sequences (GRGDYP, GRGDSP and KHIFSDDSSE) were coupled to 8% periodate oxidized alginate resulting in degrees of substitution between 3.9 and 6.9%. Cell adhesion studies of mouse myoblasts (C2C12) and human dental stem cells (RP89) to gels containing various amounts of GRGDSP coupled alginate demonstrated the bioactivity of the material where RP89 cells needed higher peptide concentrations to adhere. Copyright © 2015 Elsevier Ltd. All rights reserved.

  16. Efficacy of lactoferricin B in controlling ready-to-eat vegetable spoilage caused by Pseudomonas spp.

    Science.gov (United States)

    Federico, Baruzzi; Pinto, Loris; Quintieri, Laura; Carito, Antonia; Calabrese, Nicola; Caputo, Leonardo

    2015-12-23

    The microbial content of plant tissues has been reported to cause the spoilage of ca. 30% of chlorine-disinfected fresh vegetables during cold storage. The aim of this work was to evaluate the efficacy of antimicrobial peptides in controlling microbial vegetable spoilage under cold storage conditions. A total of 48 bacterial isolates were collected from ready-to-eat (RTE) vegetables and identified as belonging to Acinetobacter calcoaceticus, Aeromonas media, Pseudomonas cichorii, Pseudomonas fluorescens, Pseudomonas jessenii, Pseudomonas koreensis, Pseudomonas putida, Pseudomonas simiae and Pseudomonas viridiflava species. Reddish or brownish pigmentation was found when Pseudomonas strains were inoculated in wounds on leaves of Iceberg and Trocadero lettuce and escarole chicory throughout cold storage. Bovine lactoferrin (BLF) and its hydrolysates (LFHs) produced by pepsin, papain and rennin, were assayed in vitro against four Pseudomonas spp. strains selected for their heavy spoiling ability. As the pepsin-LFH showed the strongest antimicrobial effect, subsequent experiments were carried out using the peptide lactoferricin B (LfcinB), well known to be responsible for its antimicrobial activity. LfcinB significantly reduced (P ≤ 0.05) spoilage by a mean of 36% caused by three out of four inoculated spoiler pseudomonads on RTE lettuce leaves after six days of cold storage. The reduction in the extent of spoilage was unrelated to viable cell density in the inoculated wounds. This is the first paper providing direct evidence regarding the application of an antimicrobial peptide to control microbial spoilage affecting RTE leafy vegetables during cold storage.

  17. 3D Cell Culture in Alginate Hydrogels

    Directory of Open Access Journals (Sweden)

    Therese Andersen

    2015-03-01

    Full Text Available This review compiles information regarding the use of alginate, and in particular alginate hydrogels, in culturing cells in 3D. Knowledge of alginate chemical structure and functionality are shown to be important parameters in design of alginate-based matrices for cell culture. Gel elasticity as well as hydrogel stability can be impacted by the type of alginate used, its concentration, the choice of gelation technique (ionic or covalent, and divalent cation chosen as the gel inducing ion. The use of peptide-coupled alginate can control cell–matrix interactions. Gelation of alginate with concomitant immobilization of cells can take various forms. Droplets or beads have been utilized since the 1980s for immobilizing cells. Newer matrices such as macroporous scaffolds are now entering the 3D cell culture product market. Finally, delayed gelling, injectable, alginate systems show utility in the translation of in vitro cell culture to in vivo tissue engineering applications. Alginate has a history and a future in 3D cell culture. Historically, cells were encapsulated in alginate droplets cross-linked with calcium for the development of artificial organs. Now, several commercial products based on alginate are being used as 3D cell culture systems that also demonstrate the possibility of replacing or regenerating tissue.

  18. The Pseudomonas community in metal-contaminated sediments as revealed by quantitative PCR: a link with metal bioavailability.

    Science.gov (United States)

    Roosa, Stéphanie; Wauven, Corinne Vander; Billon, Gabriel; Matthijs, Sandra; Wattiez, Ruddy; Gillan, David C

    2014-10-01

    Pseudomonas bacteria are ubiquitous Gram-negative and aerobic microorganisms that are known to harbor metal resistance mechanisms such as efflux pumps and intracellular redox enzymes. Specific Pseudomonas bacteria have been quantified in some metal-contaminated environments, but the entire Pseudomonas population has been poorly investigated under these conditions, and the link with metal bioavailability was not previously examined. In the present study, quantitative PCR and cell cultivation were used to monitor and characterize the Pseudomonas population at 4 different sediment sites contaminated with various levels of metals. At the same time, total metals and metal bioavailability (as estimated using an HCl 1 m extraction) were measured. It was found that the total level of Pseudomonas, as determined by qPCR using two different genes (oprI and the 16S rRNA gene), was positively and significantly correlated with total and HCl-extractable Cu, Co, Ni, Pb and Zn, with high correlation coefficients (>0.8). Metal-contaminated sediments featured isolates of the Pseudomonas putida, Pseudomonas fluorescens, Pseudomonas lutea and Pseudomonas aeruginosa groups, with other bacterial genera such as Mycobacterium, Klebsiella and Methylobacterium. It is concluded that Pseudomonas bacteria do proliferate in metal-contaminated sediments, but are still part of a complex community. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  19. Antimicrobial resistance of Pseudomonas spp. isolated from wastewater and wastewater-impacted marine coastal zone.

    Science.gov (United States)

    Luczkiewicz, Aneta; Kotlarska, Ewa; Artichowicz, Wojciech; Tarasewicz, Katarzyna; Fudala-Ksiazek, Sylwia

    2015-12-01

    In this study, species distribution and antimicrobial susceptibility of cultivated Pseudomonas spp. were studied in influent (INF), effluent (EFF), and marine outfall (MOut) of wastewater treatment plant (WWTP). The susceptibility was tested against 8 antimicrobial classes, active against Pseudomonas spp.: aminoglycosides, carbapenems, broad-spectrum cephalosporins from the 3rd and 4th generation, extended-spectrum penicillins, as well as their combination with the β-lactamase inhibitors, monobactams, fluoroquinolones, and polymyxins. Among identified species, resistance to all antimicrobials but colistin was shown by Pseudomonas putida, the predominant species in all sampling points. In other species, resistance was observed mainly against ceftazidime, ticarcillin, ticarcillin-clavulanate, and aztreonam, although some isolates of Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas pseudoalcaligenes, and Pseudomonas protegens showed multidrug-resistance (MDR) phenotype. Among P. putida, resistance to β-lactams and to fluoroquinolones as well as multidrug resistance become more prevalent after wastewater treatment, but the resistance rate decreased in marine water samples. Obtained data, however, suggests that Pseudomonas spp. are equipped or are able to acquire a wide range of antibiotic resistance mechanisms, and thus should be monitored as possible source of resistance genes.

  20. Cytokinin production by Pseudomonas fluorescens G20-18 determines biocontrol activity against Pseudomonas syringae in Arabidopsis

    Czech Academy of Sciences Publication Activity Database

    Grosskinsky, D. K.; Tafner, R.; Moreno, M. V.; Stenglein, S. A.; Garcia de Salamone, I. E.; Nelson, L. M.; Novák, Ondřej; Strnad, Miroslav; van der Graaff, E.; Roitsch, Thomas

    2016-01-01

    Roč. 6, MAR 17 (2016), s. 23310 ISSN 2045-2322 R&D Projects: GA MŠk(CZ) LO1204; GA ČR GA15-22322S; GA MŠk(CZ) LO1415 Institutional support: RVO:61389030 ; RVO:67179843 Keywords : GROWTH-PROMOTING RHIZOBACTERIA * PLANT-GROWTH * SALICYLIC-ACID Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 4.259, year: 2016

  1. Thermostable Alginate degrading enzymes and their methods of use

    NARCIS (Netherlands)

    Hreggvidsson, Gudmundur Oli; Jonsson, Oskar W.J.; Bjornsdottir, Bryndis; Fridjonsson, Hedinn O; Altenbuchner, Josef; Watzlawick, Hildegard; Dobruchowska, Justyna; Kamerling, Johannis

    2015-01-01

    The present invention relates to the identification, production and use of thermostable alginate lyase enzymes that can be used to partially degrade alginate to yield oligosaccharides or to give complete degradation of alginate to yield (unsaturated) mono-uronates.

  2. Anti-pseudomona and Anti-bacilli Activity of Some Medicinal Plants of Iran

    Directory of Open Access Journals (Sweden)

    Gholam Hosein Shahidi Bonjar

    2003-10-01

    Full Text Available The use of plants in treatment of burns, dermatophytes, and infectious diseases is common in traditional medicine of Iran. Based on ethno pharmacological and taxonomic information, antibacterial activities of methanol extracts of some medicinal plants of Iran were determined by In Vitro bioassays using agar diffusion-method against standard strains of Pseudomonas aeruginosa, P. fluorescens, Bacillus subtilis, B. cereus and B. pumilis at 20 mg/ml. From 180 plant species of 72 families, 78 species (43.3% in 42 families (58.3% showed antibacterial activities against B. cereus (88.4%, B. subtilis (39.7%, B. pumilis (37.1%, P. fluorescens (37.1% and P. aeruginos (10.2%. The most active plant families were Apiaceae, Compositae and Labiatae with 9, 8 and 7 active plant species respectively. Minimum inhibitory concentrations (MIC of the active plants were determined using two fold serial dilutions. Most active plant against Bacilli was Myrtus communis L. with MIC of 1.87 mg/ml. For Pseudomonas species, Dianthus caryophyllus L. and Terminalia chebula (Gaertner Retz. were more active with the MIC of 0.46 mg/ml for P. fluorescens and of 1.87 mg/ml for P. aeruginosa respectively.

  3. Structural and Mechanistic Insights into the Pseudomonas fluorescens 2-Nitrobenzoate 2-Nitroreductase NbaA.

    Science.gov (United States)

    Kim, Yong-Hak; Song, Wooseok; Kim, Jin-Sik; Jiao, Li; Lee, Kangseok; Ha, Nam-Chul

    2015-08-01

    The bacterial 2-nitroreductase NbaA is the primary enzyme initiating the degradation of 2-nitrobenzoate (2-NBA), and its activity is controlled by posttranslational modifications. To date, the structure of NbaA remains to be elucidated. In this study, the crystal structure of a Cys194Ala NbaA mutant was determined to a 1.7-Å resolution. The substrate analog 2-NBA methyl ester was used to decipher the substrate binding site by inhibition of the wild-type NbaA protein. Tandem mass spectrometry showed that 2-NBA methyl ester produced a 2-NBA ester bond at the Tyr193 residue in the wild-type NbaA but not residues in the Tyr193Phe mutant. Moreover, covalent binding of the 2-NBA methyl ester to Tyr193 reduced the reactivity of the Cys194 residue on the peptide link. The Tyr193 hydroxyl group was shown to be essential for enzyme catalysis, as a Tyr193Phe mutant resulted in fast dissociation of flavin mononucleotide (FMN) from the protein with the reduced reactivity of Cys194. FMN binding to NbaA varied with solution NaCl concentration, which was related to the catalytic activity but not to cysteine reactivity. These observations suggest that the Cys194 reactivity is negatively affected by a posttranslational modification of the adjacent Tyr193 residue, which interacts with FMN and the substrate in the NbaA catalytic site. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  4. Impact of Medium on the Development and Physiology of Pseudomonas fluorescens Biofilms on Polyurethane Paint

    Science.gov (United States)

    2012-02-01

    can provide nutrients that affect biofilm formation and biodeterioration. Historically, studies of polyurethane biodeterioration have monitored the...microbial growth can accelerate the degradation of fuel tank coatings and cause corrosion of aluminum and stainless steel tanks and pipelines (Passman...conditions are determinants of both biofilm formation /dispersion and the degradative impact of bacteria on PU substrates. For example, the transition of

  5. Bistability in a Metabolic Network Underpins the De Novo Evolution of Colony Switching in Pseudomonas fluorescens

    DEFF Research Database (Denmark)

    Gallie, Jenna; Libby, Eric; Bertels, Frederic

    2015-01-01

    Phenotype switching is commonly observed in nature. This prevalence has allowed the elucidation of a number of underlying molecular mechanisms. However, little is known about how phenotypic switches arise and function in their early evolutionary stages. The first opportunity to provide empirical ...

  6. Lipopeptide biosurfactant viscosin enhances dispersal of Pseudomonas fluorescens SBW25 biofilms

    DEFF Research Database (Denmark)

    Bonnichsen, Lise; Svenningsen, Nanna Bygvraa; Rybtke, Morten Levin

    2015-01-01

    Pseudomonads produce several lipopeptide biosurfactants that have antimicrobial properties but that also facilitate surface motility and influence biofilm formation. Detailed studies addressing the significance of lipopeptides for biofilm formation and architecture are rare. Hence, the present...

  7. 76 FR 52871 - Pseudomonas fluorescens Strain CL145A; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2011-08-24

    ... factors such as ultraviolet light, nutrient depletion and bacterial grazing/predation by protists and...., environmental factors such as ultraviolet light, nutrient depletion and bacterial grazing/predation by protists...

  8. Interaction of the psychrotroph Pseudomonas fluorescens In5 with phytopathogens in cold soils

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Olsson, Stefan; Stougaard, Peter

    Aim: Potato cultivation in southwest Greenland, at Inneruulalik, omits the use of pesticides while relying on limited crop rotations and despite the presence of plant pathogens in the soil does not suffer from major disease outbreaks. Previously, we have shown that the soil at Inneruulalik...... growth phase and upon physical encounter with fungal hyphae. Furthermore, the regulator is strongly upregulated in response to carbon sources indicating the presence of a fungus suggesting that environmental elicitors may also influence regulator expression, which upon activation controls nunamycin...... temperatures could lead to increased problems with soil-borne pathogens and importantly the application of synthetic fungicides in cold areas may be problematic due to slow degradation. Thus, the application of environmentally friendly biological control agents may provide an alternative sustainable solution...

  9. Radiation degradation of alginate and some results of biological effect of degraded alginate on plants

    International Nuclear Information System (INIS)

    Hien, N.Q.; Hai, L.; Luan, L.Q.; Hanh, T.T.; Nagasawa, Naotsugu; Yoshii, Fumio; Makuuchi, Keizo; Kume, Tamikazu

    2000-01-01

    Radiation degradation yields (Gd) of alginate in aqueous solution with different concentration were determined by viscometry method. The relationship between Gd and the alginate concentration was found out as: Gd=33.5 x C -0.68 , with C% (w/v) and dry alginate referred to C=100%. An empirical equation for preparing degraded alginate with the desired low viscometry average molecular weight (Mv) by radiation was proposed. Alginate extracted directly horn seaweed'Sagassum, degraded by radiation was used for field experiments and results of the biological effect on plants (tea, carrot, chrysanthemum) were presented. (author)

  10. Radiation degradation of alginate and some results of biological effect of degraded alginate on plants

    Energy Technology Data Exchange (ETDEWEB)

    Hien, N.Q.; Hai, L.; Luan, L.Q.; Hanh, T.T. [Nuclear Research Institute, Dalat (Viet Nam); Nagasawa, Naotsugu; Yoshii, Fumio; Makuuchi, Keizo; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-03-01

    Radiation degradation yields (Gd) of alginate in aqueous solution with different concentration were determined by viscometry method. The relationship between Gd and the alginate concentration was found out as: Gd=33.5 x C{sup -0.68}, with C% (w/v) and dry alginate referred to C=100%. An empirical equation for preparing degraded alginate with the desired low viscometry average molecular weight (Mv) by radiation was proposed. Alginate extracted directly horn seaweed'Sagassum, degraded by radiation was used for field experiments and results of the biological effect on plants (tea, carrot, chrysanthemum) were presented. (author)

  11. Antimicrobial properties of Pseudomonas strains producing the antibiotic mupirocin.

    Science.gov (United States)

    Matthijs, Sandra; Vander Wauven, Corinne; Cornu, Bertrand; Ye, Lumeng; Cornelis, Pierre; Thomas, Christopher M; Ongena, Marc

    2014-10-01

    Mupirocin is a polyketide antibiotic with broad antibacterial activity. It was isolated and characterized about 40 years ago from Pseudomonas fluorescens NCIMB 10586. To study the phylogenetic distribution of mupirocin producing strains in the genus Pseudomonas a large collection of Pseudomonas strains of worldwide origin, consisting of 117 Pseudomonas type strains and 461 strains isolated from different biological origins, was screened by PCR for the mmpD gene of the mupirocin gene cluster. Five mmpD(+) strains from different geographic and biological origin were identified. They all produced mupirocin and were strongly antagonistic against Staphylococcus aureus. Phylogenetic analysis showed that mupirocin production is limited to a single species. Inactivation of mupirocin production leads to complete loss of in vitro antagonism against S. aureus, except on certain iron-reduced media where the siderophore pyoverdine is responsible for the in vitro antagonism of a mupirocin-negative mutant. In addition to mupirocin some of the strains produced lipopeptides of the massetolide group. These lipopeptides do not play a role in the observed in vitro antagonism of the mupirocin producing strains against S. aureus. Copyright © 2014 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  12. Diffusion Retardation by Binding of Tobramycin in an Alginate Biofilm Model

    DEFF Research Database (Denmark)

    Cao, Bao; Christophersen, Lars; Kolpen, Mette

    2016-01-01

    Microbial cells embedded in a self-produced extracellular biofilm matrix cause chronic infections, e. g. by Pseudomonas aeruginosa in the lungs of cystic fibrosis patients. The antibiotic killing of bacteria in biofilms is generally known to be reduced by 100–1000 times relative to planktonic...... bacteria. This makes such infections difficult to treat. We have therefore proposed that biofilms can be regarded as an independent compartment with distinct pharmacokinetics. To elucidate this pharmacokinetics we have measured the penetration of the tobramycin into seaweed alginate beads which serve...... to be uniformly distributed throughout the volume of the alginate bead. The power-law appears to be a consequence of binding to a multitude of different binding sites. In a diffusion model these results are shown to produce pronounced retardation of the penetration of tobramycin into the biofilm. This filtering...

  13. Pseudomonas biofilm matrix composition and niche biology

    Science.gov (United States)

    Mann, Ethan E.; Wozniak, Daniel J.

    2014-01-01

    Biofilms are a predominant form of growth for bacteria in the environment and in the clinic. Critical for biofilm development are adherence, proliferation, and dispersion phases. Each of these stages includes reinforcement by, or modulation of, the extracellular matrix. Pseudomonas aeruginosa has been a model organism for the study of biofilm formation. Additionally, other Pseudomonas species utilize biofilm formation during plant colonization and environmental persistence. Pseudomonads produce several biofilm matrix molecules, including polysaccharides, nucleic acids, and proteins. Accessory matrix components shown to aid biofilm formation and adaptability under varying conditions are also produced by pseudomonads. Adaptation facilitated by biofilm formation allows for selection of genetic variants with unique and distinguishable colony morphology. Examples include rugose small-colony variants and wrinkly spreaders (WS), which over produce Psl/Pel or cellulose, respectively, and mucoid bacteria that over produce alginate. The well-documented emergence of these variants suggests that pseudomonads take advantage of matrix-building subpopulations conferring specific benefits for the entire population. This review will focus on various polysaccharides as well as additional Pseudomonas biofilm matrix components. Discussions will center on structure–function relationships, regulation, and the role of individual matrix molecules in niche biology. PMID:22212072

  14. Formulation of Sodium Alginate Nanospheres Containing ...

    African Journals Online (AJOL)

    Purpose: The aim of this work was to formulate sodium alginate nanospheres of amphotericin B by controlled gellification method and to evaluate the role of the nanospheres as a “passive carrier” in targeted antifungal therapy. Methods: Sodium alginate nanospheres of amphotericin B were prepared by controlled ...

  15. Drying and Rehydration of Calcium Alginate Gels

    NARCIS (Netherlands)

    Vreeker, R.; Li, L.; Fang, Y.; Appelqvist, I.; Mendes, E.

    2008-01-01

    In this paper, we study the rehydration properties of air-dried calcium alginate gel beads. Rehydration is shown to depend on alginate source (i.e. mannuronic to guluronic acid ratio) and the salt concentration in the rehydration medium. Rehydration curves are described adequately by the empirical

  16. 21 CFR 184.1187 - Calcium alginate.

    Science.gov (United States)

    2010-04-01

    ... ingredient is used in food only within the following specific limitations: Category of food Maximum level of... other food categories 0.3 Do. (d) Prior sanctions for calcium alginate different from the uses... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Calcium alginate. 184.1187 Section 184.1187 Food...

  17. Spoilage potential of Pseudomonas species isolated from goat milk.

    Science.gov (United States)

    Scatamburlo, T M; Yamazi, A K; Cavicchioli, V Q; Pieri, F A; Nero, L A

    2015-02-01

    Pseudomonas spp. are usually associated with spoilage microflora of dairy products due to their proteolytic potential. This is of particular concern for protein-based products, such as goat milk cheeses and fermented milks. Therefore, the goal of the present study was to characterize the proteolytic activity of Pseudomonas spp. isolated from goat milk. Goat milk samples (n=61) were obtained directly from bulk tanks on dairy goat farms (n=12), and subjected to a modified International Organization for Standardization (ISO) protocol to determine the number and proteolytic activity of Pseudomonas spp. Isolates (n=82) were obtained, identified by PCR, and subjected to pulsed-field gel electrophoresis with XbaI macro-restriction. Then, the isolates were subjected to PCR to detect the alkaline protease gene (apr), and phenotypic tests were performed to check proteolytic activity at 7°C, 25°C, and 35°C. Mean Pseudomonas spp. counts ranged from 2.9 to 4.8 log cfu/mL, and proteolytic Pseudomonas spp. counts ranged from 1.9 to 4.6 log cfu/mL. All isolates were confirmed to be Pseudomonas spp., and 41 were identified as Pseudomonas fluorescens, which clustered into 5 groups sharing approximately 82% similarity. Thirty-six isolates (46.9%) were positive for the apr gene; and 57 (69.5%) isolates presented proteolytic activity at 7°C, 82 (100%) at 25°C, and 64 (78%) at 35°C. The isolates were distributed ubiquitously in the goat farms, and no relationship among isolates was observed when the goat farms, presence of apr, pulsotypes, and proteolytic activity were taken into account. We demonstrated proteolytic activity of Pseudomonas spp. present in goat milk by phenotypic and genotypic tests and indicated their spoilage potential at distinct temperatures. Based on these findings and the ubiquity of Pseudomonas spp. in goat farm environments, proper monitoring and control of Pseudomonas spp. during production are critical. Copyright © 2015 American Dairy Science Association

  18. Development of silver nanoparticles loaded chitosan-alginate constructs with biomedical potentialities.

    Science.gov (United States)

    Bilal, Muhammad; Rasheed, Tahir; Iqbal, Hafiz M N; Li, Chuanlong; Hu, Hongbo; Zhang, Xuehong

    2017-12-01

    Herein, a facile biosynthesis of silver nanoparticles (AgNPs) and AgNPs-loaded chitosan-alginate constructs with biomedical potentialities is reported. The UV-vis spectroscopic profile confirmed the synthesis of AgNPs using methanolic leaves extract of Euphorbia helioscopia. The newly developed AgNPs were characterized using various analytical and imaging techniques including UV-vis and FT-IR spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS), atomic force microscopy (AFM), and transmission electron microscopy (TEM). The optimally yielded AgNPs at 24h reaction period were loaded onto various chitosan-alginate constructs. A maximum of 95% loading efficiency (LE) was recorded with a chitosan: alginate ratio at 2:1, followed by 81% at 2:2 ratios. The anti-bacterial activities of AgNPs and AgNPs loaded chitosan-alginate constructs were tested against six bacterial strains i.e. Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, Morganella morganii and Haemophilus influenza. A significant reduction in the log values was recorded for all test constructs, in comparison to the initial bacterial count (control value, i.e., 1.5×10 8 CFU/mL). The cytotoxicity profile revealed complete biocompatibility against normal cell line i.e. L929. Almost all constructs showed considerable cytotoxicity up to certain extant against human epithelial cells (HeLa) cancer cells. In summary, the highest antibacterial activities along with anti-cancer behavior both suggest the biomedical potentialities of newly engineered AgNPs and AgNPs-loaded chitosan-alginate constructs. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Radiation degradation of alginate and chitosan

    Energy Technology Data Exchange (ETDEWEB)

    Nagasawa, Naotsugu; Mitomo, Hiroshi [Department of Biological and Chemical Engineering, Faculty of Engineering, Gunma University, Kiryu, Gunma (Japan); Yoshii, Fumio; Kume, Tamikazu [Japan Atomic Energy Research Inst., Takasaki, Gunma (Japan). Takasaki Radiation Chemistry Research Establishment

    2000-03-01

    Alginate and chitosan were irradiated in solid or aqueous solution condition with Co{sup 60} gamma rays in the dose range of 20 to 500 kGy. Degradation was observed both in solid and solution conditions. The degradation in solution was remarkably greater than that in solid. For example, the molecular weight of alginate in 4%(w/v) solution decreased from 2 x 10{sup 5} for 0 kGy to 6 x 10{sup 3} for 50 kGy irradiation while the equivalent degradation by solid irradiation required 500 kGy. The activated species from irradiated water must be responsible for the degradation in solution. The degradation was also accompanied with the color change of alginate: the color became deep brown for highly degraded alginate. UV spectra showed a distinct absorption peak at 265 nm for colored alginates, increasing with dose. The fact that discoloration of colored alginate was caused on exposure to ozone suggests a formation of double bond in pyranose-ring by scission of glycosidic bond. Degradation behavior of chitosan in irradiation was almost the same as that of alginate. (author)

  20. Modeling and Re-Engineering of Azotobacter vinelandii Alginate Lyase to Enhance Its Catalytic Efficiency for Accelerating Biofilm Degradation.

    Directory of Open Access Journals (Sweden)

    Chul Ho Jang

    Full Text Available Alginate is known to prevent elimination of Pseudomonas aeruginosa biofilms. Alginate lyase (AlgL might therefore facilitate treatment of Pseudomonas aeruginosa-infected cystic fibrosis patients. However, the catalytic activity of wild-type AlgL is not sufficiently high. Therefore, molecular modeling and site-directed mutagenesis of AlgL might assist in enzyme engineering for therapeutic development. AlgL, isolated from Azotobacter vinelandii, catalyzes depolymerization of alginate via a β-elimination reaction. AlgL was modeled based on the crystal structure template of Sphingomonas AlgL species A1-III. Based on this computational analysis, AlgL was subjected to site-directed mutagenesis to improve its catalytic activity. The kcat/Km of the K194E mutant showed a nearly 5-fold increase against the acetylated alginate substrate, as compared to the wild-type. Double and triple mutants (K194E/K245D, K245D/K319A, K194E/K245D/E312D, and K194E/K245D/K319A were also prepared. The most potent mutant was observed to be K194E/K245D/K319A, which has a 10-fold improved kcat value (against acetylated alginate compared to the wild-type enzyme. The antibiofilm effect of both AlgL forms was identified in combination with piperacillin/tazobactam (PT and the disruption effect was significantly higher in mutant AlgL combined with PT than wild-type AlgL. However, for both the wild-type and K194E/K245D/K319A mutant, the use of the AlgL enzyme alone did not show significant antibiofilm effect.

  1. Modeling and Re-Engineering of Azotobacter vinelandii Alginate Lyase to Enhance Its Catalytic Efficiency for Accelerating Biofilm Degradation.

    Science.gov (United States)

    Jang, Chul Ho; Piao, Yu Lan; Huang, Xiaoqin; Yoon, Eun Jeong; Park, So Hee; Lee, Kyoung; Zhan, Chang-Guo; Cho, Hoon

    2016-01-01

    Alginate is known to prevent elimination of Pseudomonas aeruginosa biofilms. Alginate lyase (AlgL) might therefore facilitate treatment of Pseudomonas aeruginosa-infected cystic fibrosis patients. However, the catalytic activity of wild-type AlgL is not sufficiently high. Therefore, molecular modeling and site-directed mutagenesis of AlgL might assist in enzyme engineering for therapeutic development. AlgL, isolated from Azotobacter vinelandii, catalyzes depolymerization of alginate via a β-elimination reaction. AlgL was modeled based on the crystal structure template of Sphingomonas AlgL species A1-III. Based on this computational analysis, AlgL was subjected to site-directed mutagenesis to improve its catalytic activity. The kcat/Km of the K194E mutant showed a nearly 5-fold increase against the acetylated alginate substrate, as compared to the wild-type. Double and triple mutants (K194E/K245D, K245D/K319A, K194E/K245D/E312D, and K194E/K245D/K319A) were also prepared. The most potent mutant was observed to be K194E/K245D/K319A, which has a 10-fold improved kcat value (against acetylated alginate) compared to the wild-type enzyme. The antibiofilm effect of both AlgL forms was identified in combination with piperacillin/tazobactam (PT) and the disruption effect was significantly higher in mutant AlgL combined with PT than wild-type AlgL. However, for both the wild-type and K194E/K245D/K319A mutant, the use of the AlgL enzyme alone did not show significant antibiofilm effect.

  2. Methods of detecting and controlling mucoid Pseudomonas biofilm production

    Science.gov (United States)

    Yu, Hongwei D. (Inventor); Qiu, Dongru (Inventor)

    2013-01-01

    Compositions and methods for detecting and controlling the conversion to mucoidy in Pseudomonas aeruginosa are disclosed. The present invention provides for detecting the switch from nonmucoid to mucoid state of P. aeruginosa by measuring mucE expression or MucE protein levels. The interaction between MucE and AlgW controls the switch to mucoidy in wild type P. aeruginosa. Also disclosed is an alginate biosynthesis heterologous expression system for use in screening candidate substances that inhibit conversion to mucoidy.

  3. Different Ancestries of R Tailocins in Rhizospheric Pseudomonas Isolates

    Science.gov (United States)

    Ghequire, Maarten G.K.; Dillen, Yörg; Lambrichts, Ivo; Proost, Paul; Wattiez, Ruddy; De Mot, René

    2015-01-01

    Bacterial genomes accommodate a variety of mobile genetic elements, including bacteriophage-related clusters that encode phage tail-like protein complexes playing a role in interactions with eukaryotic or prokaryotic cells. Such tailocins are unable to replicate inside target cells due to the lack of a phage head with associated DNA. A subset of tailocins mediate antagonistic activities with bacteriocin-like specificity. Functional characterization of bactericidal tailocins of two Pseudomonas putida rhizosphere isolates revealed not only extensive similarity with the tail assembly module of the Pseudomonas aeruginosa R-type pyocins but also differences in genomic integration site, regulatory genes, and lytic release modules. Conversely, these three features are quite similar between strains of the P. putida and Pseudomonas fluorescens clades, although phylogenetic analysis of tail genes suggests them to have evolved separately. Unlike P. aeruginosa R pyocin elements, the tailocin gene clusters of other pseudomonads frequently carry cargo genes, including bacteriocins. Compared with P. aeruginosa, the tailocin tail fiber sequences that act as specificity determinants have diverged much more extensively among the other pseudomonad species, mostly isolates from soil and plant environments. Activity of the P. putida antibacterial particles requires a functional lipopolysaccharide layer on target cells, but contrary to R pyocins from P. aeruginosa, strain susceptibilities surpass species boundaries. PMID:26412856

  4. Biosurfactant production by Pseudomonas strains isolated from floral nectar.

    Science.gov (United States)

    Ben Belgacem, Z; Bijttebier, S; Verreth, C; Voorspoels, S; Van de Voorde, I; Aerts, G; Willems, K A; Jacquemyn, H; Ruyters, S; Lievens, B

    2015-06-01

    To screen and identify biosurfactant-producing Pseudomonas strains isolated from floral nectar; to characterize the produced biosurfactants; and to investigate the effect of different carbon sources on biosurfactant production. Four of eight nectar Pseudomonas isolates were found to produce biosurfactants. Phylogenetic analysis based on three housekeeping genes (16S rRNA gene, rpoB and gyrB) classified the isolates into two groups, including one group closely related to Pseudomonas fluorescens and another group closely related to Pseudomonas fragi and Pseudomonas jessenii. Although our nectar pseudomonads were able to grow on a variety of water-soluble and water-immiscible carbon sources, surface active agents were only produced when using vegetable oil as sole carbon source, including olive oil, sunflower oil or waste frying sunflower oil. Structural characterization based on thin layer chromatography (TLC) and ultra high performance liquid chromatography-accurate mass mass spectrometry (UHPLC-amMS) revealed that biosurfactant activity was most probably due to the production of fatty acids (C16:0; C18:0; C18:1 and C18:2), and mono- and diglycerides thereof. Four biosurfactant-producing nectar pseudomonads were identified. The active compounds were identified as fatty acids (C16:0; C18:0; C18:1 and C18:2), and mono- and diglycerides thereof, produced by hydrolysis of triglycerides of the feedstock. Studies on biosurfactant-producing micro-organisms have mainly focused on microbes isolated from soils and aquatic environments. Here, for the first time, nectar environments were screened as a novel source for biosurfactant producers. As nectars represent harsh environments with high osmotic pressure and varying pH levels, further screening of nectar habitats for biosurfactant-producing microbes may lead to the discovery of novel biosurfactants with broad tolerance towards different environmental conditions. © 2015 The Society for Applied Microbiology.

  5. Pseudomonas aeruginosa PAO1 exopolysaccharides are important for mixed species biofilm community development and stress tolerance

    Directory of Open Access Journals (Sweden)

    Saravanan ePeriasamy

    2015-08-01

    Full Text Available Pseudomonas aeruginosa PAO1 produces three polysaccharides, alginate, Psl and Pel that play distinct roles in attachment and biofilm formation for monospecies biofilms. Considerably less is known about their role in the development of mixed species biofilm communities. This study has investigated the roles of alginate, Psl and Pel during biofilm formation of P. aeruginosa in a defined and experimentally informative mixed species biofilm community, consisting of P. aeruginosa, Pseudomonas protegens and Klebsiella pneumoniae. Loss of the Psl polysaccharide had the biggest impact on the integration of P. aeruginosa in the mixed species biofilms, where the percent composition of the psl mutant was significantly lower (0.06% than its wild-type parent (2.44%. In contrast, loss of the Pel polysaccharide had no impact on mixed species biofilm development. Loss of alginate or its overproduction resulted in P. aeruginosa representing 8.4% and 18.11%, respectively, of the mixed species biofilm. Dual species biofilms of P. aeruginosa and K. pneumoniae were not affected by loss of alginate, Pel or Psl, while the mucoid P. aeruginosa strain achieved a greater biomass than its parent strain. When P. aeruginosa was grown with P. protegens, loss of the Pel or alginate polysaccharides resulted in biofilms that were not significantly different from biofilms formed by the wild-type PAO1. In contrast, overproduction of alginate resulted in biofilms that were comprised of 35-40% of P. aeruginosa, which was significantly higher than the wild-type (5-20%. Loss of the Psl polysaccharide significantly reduced the percentage composition of P. aeruginosa in dual species biofilms with P. protegens (<1%. Loss of the Psl polysaccharide significantly disrupted the communal stress resistance of the three species biofilms. Thus, the polysaccharide composition of an individual species significantly impacts mixed species biofilm development and the emergent properties of such

  6. fibrin–chitosan–sodium alginate composite sheet

    Indian Academy of Sciences (India)

    sodium alginate composite (F–C–SA) in sheet form. F–C–SA composite was prepared and characterized for its physicochemical properties like water absorption capacity, surface morphology, FTIR spectra and mechanical properties.

  7. Formulation of Sodium Alginate Nanospheres Containing ...

    African Journals Online (AJOL)

    Patrick Erah

    controlled gellification method and to evaluate the role of the nanospheres as a ... method, and the particle size analysis was carried out by scanning electron ... capacity of sodium alginate was evaluated in terms of drug to polymer ratio.

  8. fibrin–chitosan–sodium alginate composite sheet

    Indian Academy of Sciences (India)

    MS received 1 December 2011; revised 23 February 2012 ... Fibrin, a blood plasma protein, is a minor component ... chitosan–alginate PEC membrane was prepared as a wound .... ter paper followed by accurately weighing the sample. The.

  9. Study of the biological impact of Pseudomonas spp.fluoresents on hemolyphatic metabolites and histology of the digestive tract of larvae 15 migratory locust Locusta migratoria

    International Nuclear Information System (INIS)

    Oulebsir- Mohan, H.; Doumandji-Mitiche, B.

    2012-01-01

    This study allows to test the effect of entomopathogenic bacteria of Pseudomonas fluorescens bv III and Pseudomonas fluorescence bv V on the haemolymph of Locusta migratoria metabolites, namely proteins and carbohydrates as well as on the histology of the digestive system of fifth stage larvae of migratory locust Locusta migratoria. The results show an important decrease of haemolymph protein concentration compared to controls with an increase in carbohydrate concentration. Examination of histological sections of various parts of the digestive tract showed some changes in treated. (author)

  10. Vaccination promotes TH1-like inflammation and survival in chronic Pseudomonas aeruginosa pneumonia in rats

    DEFF Research Database (Denmark)

    Johansen, H K; Hougen, H P; Cryz, S J

    1995-01-01

    In a rat model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis (CF) we studied whether the inflammatory response could be altered by vaccination. Rats were immunized with either a depolymerized alginate toxin A conjugate (D-ALG toxin A), purified alginate, an O-polysacc......In a rat model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis (CF) we studied whether the inflammatory response could be altered by vaccination. Rats were immunized with either a depolymerized alginate toxin A conjugate (D-ALG toxin A), purified alginate, an O......-polysaccharide toxin A conjugate, or sterile saline. After challenge none of the rats immunized with D-ALG toxin A died, in contrast to the other two vaccine groups combined (p = 0.03). A significant reduction in the severity of the macroscopic lung inflammation was seen in rats immunized with D-ALG toxin A, compared...... predominantly PMNs (TH2-like) to a chronic-type inflammation dominated by mononuclear leukocytes (TH1-like). In accordance, the antibody titers induced by the D-ALG toxin A vaccine were not different from those of the control rats after challenge. This study identifies a possible new way of modifying...

  11. PLGA/alginate composite microspheres for hydrophilic protein delivery

    International Nuclear Information System (INIS)

    Zhai, Peng; Chen, X.B.; Schreyer, David J.

    2015-01-01

    Poly(lactic-co-glycolic acid) (PLGA) microspheres and PLGA/alginate composite microspheres were prepared by a novel double emulsion and solvent evaporation technique and loaded with bovine serum albumin (BSA) or rabbit anti-laminin antibody protein. The addition of alginate and the use of a surfactant during microsphere preparation increased the encapsulation efficiency and reduced the initial burst release of hydrophilic BSA. Confocal laser scanning microcopy (CLSM) of BSA-loaded PLGA/alginate composite microspheres showed that PLGA, alginate, and BSA were distributed throughout the depths of microspheres; no core/shell structure was observed. Scanning electron microscopy revealed that PLGA microspheres erode and degrade more quickly than PLGA/alginate composite microspheres. When loaded with anti-laminin antibody, the function of released antibody was well preserved in both PLGA and PLGA/alginate composite microspheres. The biocompatibility of PLGA and PLGA/alginate microspheres were examined using four types of cultured cell lines, representing different tissue types. Cell survival was variably affected by the inclusion of alginate in composite microspheres, possibly due to the sensitivity of different cell types to excess calcium that may be released from the calcium cross-linked alginate. - Highlights: • A double emulsion technique is used to prepare protein-loaded PLGA or PLGA/alginate microspheres. • PLGA, alginate and protein are distributed evenly within microsphere structure. • Addition of alginate improves loading efficiency and slows degradation and protein release. • PLGA/alginate microspheres have favorable biocompatibility

  12. PLGA/alginate composite microspheres for hydrophilic protein delivery

    Energy Technology Data Exchange (ETDEWEB)

    Zhai, Peng [Department of Anatomy and Cell Biology, University of Saskatchewan, S7N5E5 (Canada); Division of Biomedical Engineering, University of Saskatchewan, S7N5A9 (Canada); Chen, X.B. [Department of Mechanical Engineering, University of Saskatchewan, S7N5A9 (Canada); Division of Biomedical Engineering, University of Saskatchewan, S7N5A9 (Canada); Schreyer, David J., E-mail: david.schreyer@usask.ca [Department of Anatomy and Cell Biology, University of Saskatchewan, S7N5E5 (Canada); Division of Biomedical Engineering, University of Saskatchewan, S7N5A9 (Canada)

    2015-11-01

    Poly(lactic-co-glycolic acid) (PLGA) microspheres and PLGA/alginate composite microspheres were prepared by a novel double emulsion and solvent evaporation technique and loaded with bovine serum albumin (BSA) or rabbit anti-laminin antibody protein. The addition of alginate and the use of a surfactant during microsphere preparation increased the encapsulation efficiency and reduced the initial burst release of hydrophilic BSA. Confocal laser scanning microcopy (CLSM) of BSA-loaded PLGA/alginate composite microspheres showed that PLGA, alginate, and BSA were distributed throughout the depths of microspheres; no core/shell structure was observed. Scanning electron microscopy revealed that PLGA microspheres erode and degrade more quickly than PLGA/alginate composite microspheres. When loaded with anti-laminin antibody, the function of released antibody was well preserved in both PLGA and PLGA/alginate composite microspheres. The biocompatibility of PLGA and PLGA/alginate microspheres were examined using four types of cultured cell lines, representing different tissue types. Cell survival was variably affected by the inclusion of alginate in composite microspheres, possibly due to the sensitivity of different cell types to excess calcium that may be released from the calcium cross-linked alginate. - Highlights: • A double emulsion technique is used to prepare protein-loaded PLGA or PLGA/alginate microspheres. • PLGA, alginate and protein are distributed evenly within microsphere structure. • Addition of alginate improves loading efficiency and slows degradation and protein release. • PLGA/alginate microspheres have favorable biocompatibility.

  13. Occurrence of multi-antibiotic resistant Pseudomonas spp. in drinking water produced from karstic hydrosystems.

    Science.gov (United States)

    Flores Ribeiro, Angela; Bodilis, Josselin; Alonso, Lise; Buquet, Sylvaine; Feuilloley, Marc; Dupont, Jean-Paul; Pawlak, Barbara

    2014-08-15

    Aquatic environments could play a role in the spread of antibiotic resistance genes by enabling antibiotic-resistant bacteria transferred through wastewater inputs to connect with autochthonous bacteria. Consequently, drinking water could be a potential pathway to humans and animals for antibiotic resistance genes. The aim of this study was to investigate occurrences of Escherichia coli and Pseudomonas spp. in drinking water produced from a karst, a vulnerable aquifer with frequent increases in water turbidity after rainfall events and run-offs. Water samples were collected throughout the system from the karstic springs to the drinking water tap during three non-turbid periods and two turbid events. E. coli densities in the springs were 10- to 1000-fold higher during the turbid events than during the non-turbid periods, indicating that, with increased turbidity, surface water had entered the karstic system and contaminated the spring water. However, no E. coli were isolated in the drinking water. In contrast, Pseudomonas spp. were isolated from the drinking water only during turbid events, while the densities in the springs were from 10- to 100-fold higher than in the non-turbid periods. All the 580 Pseudomonas spp. isolates obtained from the sampling periods were resistant (to between 1 and 10 antibiotics), with similar resistance patterns. Among all the Pseudomonas isolated throughout the drinking water production system, between 32% and 86% carried the major resistance pattern: ticarcillin, ticarcillin-clavulanic acid, cefsulodin, and/or aztreonam, and/or sulfamethoxazol-trimethoprim, and/or fosfomycin. Finally, 8 Pseudomonas spp. isolates, related to the Pseudomonas putida and Pseudomonas fluorescens species, were isolated from the drinking water. Thus, Pseudomonas could be involved in the dissemination of antibiotic resistance via drinking water during critical periods. Copyright © 2014 Elsevier B.V. All rights reserved.

  14. Development of Alginate/Chitosan Microparticles for Dust Mite ...

    African Journals Online (AJOL)

    Erah

    surface of chitosan microparticles [4]. .... The reverse-phase high performance liquid .... The surface charge of alginate ... negative charge was as a result of the alginate on the microparticle surface. ... electrostatic interaction of the positively-.

  15. Characterization of initial events in bacterial surface colonization by two Pseudomonas species using image analysis.

    Science.gov (United States)

    Mueller, R F; Characklis, W G; Jones, W L; Sears, J T

    1992-05-01

    The processes leading to bacterial colonization on solid-water interfaces are adsorption, desorption, growth, and erosion. These processes have been measured individually in situ in a flowing system in real time using image analysis. Four different substrata (copper, silicon, 316 stainless-steel and glass) and 2 different bacterial species (Pseudomonas aeruginosa and Pseudomonas fluorescens) were used in the experiments. The flow was laminar (Re = 1.4) and the shear stress was kept constant during all experiments at 0.75 N m(-2). The surface roughness varied among the substrata from 0.002 microm (for silicon) to 0.015 microm (for copper). Surface free energies varied from 25.1 dynes cm(-1) for silicon to 31.2 dynes cm(-1) for copper. Cell curface hydrophobicity, reported as hydrocarbon partitioning values, ranged from 0.67 for Ps. fluorescens to 0.97 for Ps. aeruginosa.The adsorption rate coefficient varied by as much as a factor of 10 among the combinations of bacterial strain and substratum material, and was positively correlated with surface free energy, the surface roughness of the substratum, and the hydrophobicity of the cells. The probability of desorption decreased with increasing surface free energy and surface roughness of the substratum. Cell growth was inhibited on copper, but replication of cells overlying an initial cell layer was observed with increased exposure time to the cell-containing bulk water. A mathematical model describing cell accumulation on a substratum is presented.

  16. Comparative genomic and functional analyses: unearthing the diversity and specificity of nematicidal factors in Pseudomonas putida strain 1A00316

    Science.gov (United States)

    Guo, Jing; Jing, Xueping; Peng, Wen-Lei; Nie, Qiyu; Zhai, Yile; Shao, Zongze; Zheng, Longyu; Cai, Minmin; Li, Guangyu; Zuo, Huaiyu; Zhang, Zhitao; Wang, Rui-Ru; Huang, Dian; Cheng, Wanli; Yu, Ziniu; Chen, Ling-Ling; Zhang, Jibin

    2016-01-01

    We isolated Pseudomonas putida (P. putida) strain 1A00316 from Antarctica. This bacterium has a high efficiency against Meloidogyne incognita (M. incognita) in vitro and under greenhouse conditions. The complete genome of P. putida 1A00316 was sequenced using PacBio single molecule real-time (SMRT) technology. A comparative genomic analysis of 16 Pseudomonas strains revealed that although P. putida 1A00316 belonged to P. putida, it was phenotypically more similar to nematicidal Pseudomonas fluorescens (P. fluorescens) strains. We characterized the diversity and specificity of nematicidal factors in P. putida 1A00316 with comparative genomics and functional analysis, and found that P. putida 1A00316 has diverse nematicidal factors including protein alkaline metalloproteinase AprA and two secondary metabolites, hydrogen cyanide and cyclo-(l-isoleucyl-l-proline). We show for the first time that cyclo-(l-isoleucyl-l-proline) exhibit nematicidal activity in P. putida. Interestingly, our study had not detected common nematicidal factors such as 2,4-diacetylphloroglucinol (2,4-DAPG) and pyrrolnitrin in P. putida 1A00316. The results of the present study reveal the diversity and specificity of nematicidal factors in P. putida strain 1A00316. PMID:27384076

  17. Effect of Iron Availability on Induction of Systemic Resistance to Fusarium Wilt of Chickpea by Pseudomonas spp.

    Science.gov (United States)

    Saikia, Ratul; Srivastava, Alok K; Singh, Kiran; Arora, Dilip K; Lee, Min-Woong

    2005-03-01

    Selected isolates of Pseudomonas fluorescens (Pf4-92 and PfRsC5) and P. aeruginosa (PaRsG18 and PaRsG27) were examined for growth promotion and induced systemic resistance against Fusarium wilt of chickpea. Significant increase in plant height was observed in Pseudomonas treated plants. However, plant growth was inhibited when isolates of Pseudomonas were used in combination with Fusarium oxysporum f. sp. ciceri (FocRs1). It was also observed that the Pseudomonas spp. was colonized in root of chickpea and significantly suppressed the disease in greenhouse condition. Rock wool bioassay technique was used to study the effect of iron availability on the induction of systemic resistance to Fusarium wilt of chickpea mediated by the Pseudomonas spp. All the isolates of Pseudomonas spp. showed greater disease control in the induced systemic resistance (ISR) bioassay when iron availability in the nutrient solution was low. High performance liquid chromatography (HPLC) analysis indicated that all the bacterial isolates produced more salicylic acid (SA) at low iron (10µM EDDHA) than high iron availability (10µFe(3+) EDDHA). Except PaRsG27, all the three isolates produced more pseudobactin at low iron than high iron availability.

  18. Physicochemical properties of marine collagen-alginate biomaterial

    Science.gov (United States)

    Soon, K. S.; Hii, S. L.; Wong, C. L.; Leong, L. K.; Woo, K. K.

    2017-12-01

    Collagen base biomaterials are widely applied in the field of tissue engineering. However, these fibrous proteins in animal connective tissues are insufficient to fulfill the mechanical properties for such applications. Therefore, alginate as a natural polysaccharide was incorporated. In this study, Smooth wolf herring skins was collected from the local fish ball processing industry for collagen extraction using acid solubilisation method. On the other hand, alginate from brown seaweed (Sargassum polycystum) was extracted with calcium carbonate at 50 °C. The composite films of different collagen and alginate ratio were prepared by lyophilisation with pure collagen film as control. The effects of alginate on swelling behaviour, porosity, collagenase degradation and tensile strength of the composite films were investigated. Swelling behaviour increased with alginate content, 50 % alginate film achieved 1254.75 % swelling after 24 h. All composite films achieved more than 80 % porosity except the film with 80 % collagen (65.41 %). Porosity was highest in 100 % alginate (94.30 %). Highest tensile strength (1585.87 kPa) and young modulus (27.05 MPa) was found in 50 % alginate film. In addition, resistance to collagenase degradation was improved with alginate content, lowest degradation rate was determined in 80 % alginate film. Results indicated alginate is efficient in improving some mechanical properties of the composite film.

  19. Storage duration effect on deformation recovery of repacked alginates

    Directory of Open Access Journals (Sweden)

    Siti Sunarintyas

    2009-09-01

    Full Text Available Background: Manufacturers supply alginate impression materials as a powder that is packaged in bulk and in individual container. Some Indonesian dental suppliers often repackage the bulk alginate into individual plastic packages which are not tied tightly and stored in the display room without air conditioner. It is known that critical factors to the shelf life of alginate includer avoidance of moisture contamination which may lead to premature setting of the alginate and avoidance of high temperature which may cause depolymerization of the alginate. Purpose: The aim of this study was to determine storage duration effect of repacked alginates on deformation recovery. Methods: Two brands of alginates (Tulip®TU, and Aroma Fine DF III®AF were repacked into 120 plastic containers. The samples were stored in room condition (temperature 29° C ± 1° C, relative humidity 60% ± 10% for 1, 2, 3, 4 and 5 weeks. The alginates setting time and recovery from deformation were measured according to the ANSI/ADA specification number 18 (ISO 1563. result: The results revealed that there was decreased setting time during 5 weeks but there was slight decreased in deformation recovery after 3 weeks storage. The ANOVA showed there was no significant difference of alginates deformation recovery among the storage times (p > 0.05. Conclusion: Storage duration of repacked alginates in plastic containers during 5 weeks in room condition do not influence the alginate deformation recovery.

  20. Pseudomonas Lipopeptide Biosurfactants

    DEFF Research Database (Denmark)

    Bonnichsen, Lise

    Pseudomonas lipopetide biosurfactants are amphiphilic molecules with a broad range of natural functions. Due to their surface active properties, it has been suggested that Pseudomonas lipopetides potentially play a role in biodegradation of hydrophobic compounds and have essential functions...... lipopetide biosurfactants in pollutant biodegradation and natural roles in biofilm formation. The work presented is a combination of environmental microbiology and exploiting genetic manipulation of pure cultures to achieve insightinto the effects and mechanisms of lipopeptides on microbial processes...

  1. Self-disinfecting Alginate vs Conventional Alginate: Effect on Surface Hardness of Gypsum Cast-An in vitro Study.

    Science.gov (United States)

    Madhavan, Ranjith; George, Navia; Thummala, Niharika R; Ravi, S V; Nagpal, Ajay

    2017-11-01

    For the construction of any dental prosthesis, accurate impressions are necessary. Hence, we undertook the present study to evaluate and compare the surface hardness of gypsum casts poured from impressions made using conventional alginate and self-disinfecting alginate. A total of 30 impressions of stainless steel die were made, out of which 15 impressions were made with conventional alginate and 15 were made with self-disinfecting alginate and poured using Type III dental stone. Thirty stone specimens were subjected for hardness testing. Data were analyzed using independent samples t-test to compare the mean surface hardness. Difference in surface hardness was statistically insignificant (p > 0.05). Surface hardness of gypsum casts poured using impressions made from self-disinfecting alginate and conventional alginates were comparable. Self-disinfecting alginates may be employed in clinical practice as safe and effective materials to overcome the infection control issues without compromising on the properties of the material.

  2. Development of sodium alginate and konkoli gumgrafted ...

    African Journals Online (AJOL)

    This experiment is a continuation of our effort to develop a blend membrane of sodium alginate and “konkoli” gum-g-polyacrylamide (KG-g-PAAm) for bioremediation of wastewater. The effect of graft reaction conditions on the percentage graft yield in the graft copolymerization was investigated. It was observed that grafting ...

  3. 21 CFR 184.1610 - Potassium alginate.

    Science.gov (United States)

    2010-04-01

    ...: Category of food Maximum level of use in food (as served) (percent) Functional use Confections and...) of this chapter 0.25 Do. All other food categories 0.01 Do. (d) Prior sanctions for potassium... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Potassium alginate. 184.1610 Section 184.1610 Food...

  4. 21 CFR 184.1011 - Alginic acid.

    Science.gov (United States)

    2010-04-01

    ... used in food only within the following specific limitations: Category of food Maximum level of use in... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Alginic acid. 184.1011 Section 184.1011 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) FOOD FOR HUMAN...

  5. Ferric-Pyoverdine Recognition by Fpv Outer Membrane Proteins of Pseudomonas protegens Pf-5

    Science.gov (United States)

    Hartney, Sierra L.; Mazurier, Sylvie; Girard, Maëva K.; Mehnaz, Samina; Davis, Edward W.; Gross, Harald; Lemanceau, Philippe

    2013-01-01

    The soil bacterium Pseudomonas protegens Pf-5 (previously called P. fluorescens Pf-5) produces two siderophores, enantio-pyochelin and a compound in the large and diverse pyoverdine family. Using high-resolution mass spectroscopy, we determined the structure of the pyoverdine produced by Pf-5. In addition to producing its own siderophores, Pf-5 also utilizes ferric complexes of some pyoverdines produced by other strains of Pseudomonas spp. as sources of iron. Previously, phylogenetic analysis of the 45 TonB-dependent outer membrane proteins in Pf-5 indicated that six are in a well-supported clade with ferric-pyoverdine receptors (Fpvs) from other Pseudomonas spp. We used a combination of phylogenetics, bioinformatics, mutagenesis, pyoverdine structural determinations, and cross-feeding bioassays to assign specific ferric-pyoverdine substrates to each of the six Fpvs of Pf-5. We identified at least one ferric-pyoverdine that was taken up by each of the six Fpvs of Pf-5. Functional redundancy of the Pf-5 Fpvs was also apparent, with some ferric-pyoverdines taken up by all mutants with a single Fpv deletion but not by a mutant having deletions in two of the Fpv-encoding genes. Finally, we demonstrated that phylogenetically related Fpvs take up ferric complexes of structurally related pyoverdines, thereby establishing structure-function relationships that can be employed in the future to predict the pyoverdine substrates of Fpvs in other Pseudomonas spp. PMID:23222724

  6. Antimicrobial Activity of Various Plant Extracts on Pseudomonas Species Associated with Spoilage of Chilled Fish

    Directory of Open Access Journals (Sweden)

    Osan Bahurmiz

    2016-11-01

    Full Text Available The antimicrobial activity of various plant extracts on Pseudomonas bacteria isolated from spoiled chilled tilapia (Oreochromis sp. was evaluated in this study. In the first stage of this study, red tilapia was subjected to chilled storage (4°C for 3 weeks, and spoilage bacteria were isolated and identified from the spoiled fish. Pseudomonas was the dominant bacteria isolated from the spoiled fish and further identification revealed that P. putida, P. fluorescens and Pseudomonas spp. were the main species of this group. In the second stage, methanolic extracts of 15 selected plant species were screened for their antimicrobial activity, by agar disc diffusion method, against the Pseudomonas isolates. Results indicated that most of the extracts had different degrees of activity against the bacterial isolates. The strongest activity was exhibited by bottlebrush flower (Callistemon viminalis extract. This was followed by extracts from guava bark (Psidium guajava and henna leaf (Lawsonia inermis. Moderate antimicrobial activities were observed in extracts of clove (Syzygium aromaticum, leaf and peel of tamarind (Tamarindus indica, cinnamon bark (Cinnamomum zeylanicum, wild betel leaf (Piper sarmentosum and fresh thyme (Thymus spp.. Weak or no antimicrobial activity was observed from the remaining extracts. The potential antimicrobial activity shown by some plant extracts in this study could significantly contribute to the fish preservation.

  7. Biomolecular Mechanisms of Pseudomonas aeruginosa and Escherichia coli Biofilm Formation

    Science.gov (United States)

    Laverty, Garry; Gorman, Sean P.; Gilmore, Brendan F.

    2014-01-01

    Pseudomonas aeruginosa and Escherichia coli are the most prevalent Gram-negative biofilm forming medical device associated pathogens, particularly with respect to catheter associated urinary tract infections. In a similar manner to Gram-positive bacteria, Gram-negative biofilm formation is fundamentally determined by a series of steps outlined more fully in this review, namely adhesion, cellular aggregation, and the production of an extracellular polymeric matrix. More specifically this review will explore the biosynthesis and role of pili and flagella in Gram-negative adhesion and accumulation on surfaces in Pseudomonas aeruginosa and Escherichia coli. The process of biofilm maturation is compared and contrasted in both species, namely the production of the exopolysaccharides via the polysaccharide synthesis locus (Psl), pellicle Formation (Pel) and alginic acid synthesis in Pseudomonas aeruginosa, and UDP-4-amino-4-deoxy-l-arabinose and colonic acid synthesis in Escherichia coli. An emphasis is placed on the importance of the LuxR homologue sdiA; the luxS/autoinducer-II; an autoinducer-III/epinephrine/norepinephrine and indole mediated Quorum sensing systems in enabling Gram-negative bacteria to adapt to their environments. The majority of Gram-negative biofilms consist of polysaccharides of a simple sugar structure (either homo- or heteropolysaccharides) that provide an optimum environment for the survival and maturation of bacteria, allowing them to display increased resistance to antibiotics and predation. PMID:25438014

  8. Biomolecular Mechanisms of Pseudomonas aeruginosa and Escherichia coli Biofilm Formation

    Directory of Open Access Journals (Sweden)

    Garry Laverty

    2014-07-01

    Full Text Available Pseudomonas aeruginosa and Escherichia coli are the most prevalent Gram-negative biofilm forming medical device associated pathogens, particularly with respect to catheter associated urinary tract infections. In a similar manner to Gram-positive bacteria, Gram-negative biofilm formation is fundamentally determined by a series of steps outlined more fully in this review, namely adhesion, cellular aggregation, and the production of an extracellular polymeric matrix. More specifically this review will explore the biosynthesis and role of pili and flagella in Gram-negative adhesion and accumulation on surfaces in Pseudomonas aeruginosa and Escherichia coli. The process of biofilm maturation is compared and contrasted in both species, namely the production of the exopolysaccharides via the polysaccharide synthesis locus (Psl, pellicle Formation (Pel and alginic acid synthesis in Pseudomonas aeruginosa, and UDP-4-amino-4-deoxy-l-arabinose and colonic acid synthesis in Escherichia coli. An emphasis is placed on the importance of the LuxR homologue sdiA; the luxS/autoinducer-II; an autoinducer-III/epinephrine/norepinephrine and indole mediated Quorum sensing systems in enabling Gram-negative bacteria to adapt to their environments. The majority of Gram-negative biofilms consist of polysaccharides of a simple sugar structure (either homo- or heteropolysaccharides that provide an optimum environment for the survival and maturation of bacteria, allowing them to display increased resistance to antibiotics and predation.

  9. Investigation of the algT operon sequence in mucoid and non-mucoid Pseudomonas aeruginosa isolates from 115 Scandinavian patients with cystic fibrosis and in 88 in vitro non-mucoid revertants

    DEFF Research Database (Denmark)

    Ciofu, Oana; Lee, Baoleri; Johannesson, Marie

    2008-01-01

    Pseudomonas aeruginosa is the dominant pathogen causing chronic lung infections in patients with cystic fibrosis (CF). After an initial phase characterized by intermittent colonizations, a chronic infection is established upon conversion of P. aeruginosa from the non-mucoid to the mucoid, alginate...

  10. Comparative genomic analysis of four representative plant growth-promoting rhizobacteria in Pseudomonas

    Science.gov (United States)

    2013-01-01

    Background Some Pseudomonas strains function as predominant plant growth-promoting rhizobacteria (PGPR). Within this group, Pseudomonas chlororaphis and Pseudomonas fluorescens are non-pathogenic biocontrol agents, and some Pseudomonas aeruginosa and Pseudomonas stutzeri strains are PGPR. P. chlororaphis GP72 is a plant growth-promoting rhizobacterium with a fully sequenced genome. We conducted a genomic analysis comparing GP72 with three other pseudomonad PGPR: P. fluorescens Pf-5, P. aeruginosa M18, and the nitrogen-fixing strain P. stutzeri A1501. Our aim was to identify the similarities and differences among these strains using a comparative genomic approach to clarify the mechanisms of plant growth-promoting activity. Results The genome sizes of GP72, Pf-5, M18, and A1501 ranged from 4.6 to 7.1 M, and the number of protein-coding genes varied among the four species. Clusters of Orthologous Groups (COGs) analysis assigned functions to predicted proteins. The COGs distributions were similar among the four species. However, the percentage of genes encoding transposases and their inactivated derivatives (COG L) was 1.33% of the total genes with COGs classifications in A1501, 0.21% in GP72, 0.02% in Pf-5, and 0.11% in M18. A phylogenetic analysis indicated that GP72 and Pf-5 were the most closely related strains, consistent with the genome alignment results. Comparisons of predicted coding sequences (CDSs) between GP72 and Pf-5 revealed 3544 conserved genes. There were fewer conserved genes when GP72 CDSs were compared with those of A1501 and M18. Comparisons among the four Pseudomonas species revealed 603 conserved genes in GP72, illustrating common plant growth-promoting traits shared among these PGPR. Conserved genes were related to catabolism, transport of plant-derived compounds, stress resistance, and rhizosphere colonization. Some strain-specific CDSs were related to different kinds of biocontrol activities or plant growth promotion. The GP72 genome

  11. Produção de alginato por microrganismos Alginate production by microorganisms

    Directory of Open Access Journals (Sweden)

    José Miguel Müller

    2011-01-01

    Full Text Available O alginato é um copolímero linear constituído de unidades de ácidos α-L-gulurônicos e β-D-manurônicos e é extensamente utilizado devido as suas propriedades espessantes, estabilizantes e gelificantes. Estas características fazem com que este biopolímero encontre aplicações na indústria de alimentos, na indústria têxtil e de papel, em cosméticos e na área farmacêutica e médica. Atualmente para este conjunto de aplicações sua principal fonte são algas marrons, entretanto, o alginato pode ser obtido a partir de biossíntese, utilizando-se microrganismos do gênero Pseudomonas e Azotobacter. A produção bacteriana de alginato apresenta-se como uma alternativa interessante e sua produção por microrganismos, além de possibilitar a produção de biopolímeros de alta qualidade com características específicas e pré-determinadas, irá diminuir o impacto ambiental nas regiões em que as algas marinhas das quais é extraído são coletadas. Nos últimos anos, vários estudos relacionados à produção de alginato por microrganismos foram realizados com o objetivo de avaliar sua produção e rota metabólica de biossíntese, para caracterizar o material produzido e para determinar as potencialidades de aplicação deste novo material. O rápido desenvolvimento de aplicações do alginato na área médica e farmacêutica, bem como a descoberta de propriedades imunológicas únicas deste material tem aumentado o interesse no desenvolvimento de processos para produzi-lo. Neste artigo são abordados aspectos relacionados à produção e as características do alginato bacteriano e também reportadas às potencialidades e aplicações inovadoras nas quais este material vem sendo utilizado.Alginate is a linear copolymer consisting of units of α-L-guluronic and β-D-mannuronic acid which is widely used due to its thickening, stabilizing and gelling properties. These characteristics mean that it has many applications in the food

  12. Radiation protection by ascorbic acid in sodium alginate solutions

    Energy Technology Data Exchange (ETDEWEB)

    Aliste, A.J.; Mastro, N.L. Del [Center of Radiation Technology, IPEN/CNEN/SP, University City, 05508-000 Sao Paulo (Brazil)]. E-mail: ajaliste@ipen.br

    2004-07-01

    Alginates are gelling hydrocolloids extracted from brown seaweed used widely in the nourishing and pharmaceutical industries. As alginic acid gellification retard food entrance in the stomach alginate is an additive used in diets. The objective of this work was to study the protective action of the ascorbic acid in alginate solutions against the action of {sup 60} Co gamma radiation. One % (w/v) solutions of alginate had been used and concentrations of ascorbic acid varied from 0 to 2.5% (w/v). The solutions were irradiated with doses up to 10 kGy. Viscosity/dose relationship and the p H of the solutions at 25 Centigrade were determined. Ascorbic acid behaved as an antioxidant against radiation oxidative shock in this model system of an irradiated viscous solution. Besides its radiation protective role on alginate solutions ascorbic acid promoted a viscosity increase in the range of concentrations employed. (Author)

  13. Radiation protection by ascorbic acid in sodium alginate solutions

    International Nuclear Information System (INIS)

    Aliste, A.J.; Mastro, N.L. Del

    2004-01-01

    Alginates are gelling hydrocolloids extracted from brown seaweed used widely in the nourishing and pharmaceutical industries. As alginic acid gellification retard food entrance in the stomach alginate is an additive used in diets. The objective of this work was to study the protective action of the ascorbic acid in alginate solutions against the action of 60 Co gamma radiation. One % (w/v) solutions of alginate had been used and concentrations of ascorbic acid varied from 0 to 2.5% (w/v). The solutions were irradiated with doses up to 10 kGy. Viscosity/dose relationship and the p H of the solutions at 25 Centigrade were determined. Ascorbic acid behaved as an antioxidant against radiation oxidative shock in this model system of an irradiated viscous solution. Besides its radiation protective role on alginate solutions ascorbic acid promoted a viscosity increase in the range of concentrations employed. (Author)

  14. Azithromycin blocks quorum sensing and alginate polymer formation and increases the sensitivity to serum and stationary growth phase killing of P. aeruginosa and attenuates chronic P. aeruginosa lung infection in Cftr -/--mice

    DEFF Research Database (Denmark)

    Hoffmann, N.; Lee, Bao le ri; Hentzer, Morten

    2007-01-01

    The consequences of O-acetylated alginate-producing Pseudomonas aeruginosa biofilms in the lungs of chronically infected cystic fibrosis (CF) patients are tolerance to both antibiotic treatments and effects on the innate and the adaptive defense mechanisms. In clinical trials, azithromycin (AZM...... and the complement system. Moreover, we show that AZM may affect the polymerization of P. aeruginosa alginate by the incomplete precipitation of polymerized alginate and high levels of readily dialyzable uronic acids. In addition, we find that mucoid bacteria in the stationary growth phase became sensitive to AZM......, whereas cells in the exponential phase did not. Interestingly, AZM-treated P. aeruginosa lasI mutants appeared to be particularly resistant to serum, whereas bacteria with a functional QS system did not. We show in a CF mouse model of chronic P. aeruginosa lung infection that AZM treatment results...

  15. Variations in Calcium and Alginate Ions Concentration in Relation to the Properties of Calcium Alginate Nanoparticles

    Directory of Open Access Journals (Sweden)

    Hamed Daemi

    2013-05-01

    Full Text Available Alginate belongs to a group of natural polymers called polysaccharides. They have carboxylic functional groups beside hydroxyls which are common in all polysaccharides. These materials show interesting properties due to theirfunctional groups. One of these properties is the ability of this polymer as a suitable carrier of protecting and transferring drugs and biomolecules. The particle sizes of these polymers are very important for their applications, so different techniques were used for preparation of these materials. In this way polymeric nanoparticles of calcium alginate which are excellent carriers in drug delivery systems were prepared by addition of calcium chloride solution to dilute solution of sodium alginate. Investigation of the size and distribution of nanoparticles were analyzed by SEM method. The concentration effects of both alginate and calcium ions on the size and distribution of  nanoparticles were studied in this research. Results showed that the size of nanoparticles obviously decreased with decreasing polymeric alginate concentration because of lower active sites in polymer chain. On the other hand, thesize and distribution of nanoparticles are significantly improved with increase of calcium cation concentrations. The mean particle size 40-70 nm and spherical shape are the main characteristics of the prepared nanoparticles.

  16. Cytotoxicity, Bactericidal, and Antioxidant Activity of Sodium Alginate Hydrosols Treated with Direct Electric Current

    Directory of Open Access Journals (Sweden)

    Żaneta Król

    2017-03-01

    Full Text Available The aim of the study was to investigate the effect of using direct electric current (DC of 0, 200, and 400 mA for five minutes on the physiochemical properties, cytotoxicity, antibacterial, and antioxidant activity of sodium alginate hydrosols with different sodium chloride concentrations. The pH, oxidation-reduction potential (ORP, electrical conductivity (EC, and available chlorine concentration (ACC were measured. The effect of sodium alginate hydrosols treated with DC on Staphylococcus aureus, Listeria monocytogenes, Bacillus cereus, Micrococcus luteus, Escherichia coli, Salmonella enteritidis, Yersinia enterocolitica, Pseudomonas fluorescence, and RAW 264.7 and L929 cells was investigated. Subsequently, the antioxidant properties of hydrosols were evaluated by determining the scavenging ability of 1,1-diphenyl-2-picrylhydrazyl free radical (DPPH and ferric reducing antioxidant power (FRAP. The results have shown that after applying 400 mA in hydrosol samples with 0.1% and 0.2% NaCl all tested bacteria were inactivated. The ACC concentration of C400 samples with NaCl was equal to 13.95 and 19.71 mg/L, respectively. The cytotoxicity analysis revealed that optimized electric field conditions and the addition of sodium chloride allow for the avoidance of toxicity effects on normal cells without disturbing the antibacterial effects. Due to the presence of oxidizing substances, the DPPH of variants treated with DC was lower than the DPPH of control samples.

  17. Preservation of viability and antibacterial activity of Lactobacillus spp. in calcium alginate beads.

    Science.gov (United States)

    Brachkova, Mariya I; Duarte, Maria A; Pinto, João F

    2010-12-23

    The objective of the study was to produce calcium alginate beads able to deliver Lactobacillus spp. (Lactobacillus plantarum, Lactobacillus rhamnosus GG, Lactobacillus bulgaricus and Lactobacillus lactis) with preserved viability and antibacterial activity. Four types of beads, containing entrapped (E), surface and entrapped (ES), surface (S) and concentrated surface and entrapped lactobacilli (C(ES)) were prepared and physically characterized. The antibacterial activity of lactobacilli cultures before and after immobilization, freeze-drying and throughout storage was studied in relationship to the viable number of lactobacilli. Multi-resistant clinical isolates (methicillin-resistant Staphylococcus aureus, vancomycine-resistant Enterococcus faecalis, VIM-2-metalo-β-lactamase producing Pseudomonas aeruginosa and CTX-M-15-β-lactamase producing strains: Escherichia coli and Klebsiella pneumoniae) were used as indicator strains. Alginate beads in which lactobacilli proliferated to the beads surface (ES and C(ES)) differed significantly from the other types of beads in their physicochemical properties, showing smoother surface morphology, more spherical shape, bigger weight, lower calcium content, density and crushing force. Lactobacilli cultures, at high cell concentrations (10(8)cfu/ml) were active against both Gram-positive and negative multi-resistant bacteria. Beads containing both entrapped and surface lactobacilli (ES) resulted in viability and antibacterial activity most similar to non-processed lactobacilli cultures. The viability and antibacterial activity of the immobilized lactobacilli remained stable after 6 months storage. Copyright © 2010 Elsevier B.V. All rights reserved.

  18. Physical- chemical changes in irradiated sodium alginate algimar

    International Nuclear Information System (INIS)

    Rapado Paneque, Manuel; Alazanes, Sonia; Sainz Vidal, Dianelys; Wandrey, Christine

    2003-01-01

    The effect of gamma radiation on the physical-chemical properties of sodium alginate Algimar has been investigated. dilution viscometric, densitometry FTIR spectroscopy served to identify modifications. Decreasing intrinsic, viscosities clearly revealed chain cleavage for both solid alginate indicate that chain degradation occurs without significant change of the chemical structure, The obtained results have practical implication change of the chemical structure. The obtained results have practical implication in the field of radiation modification and sterilization of sodium alginate used for microcapsule formation

  19. THERMAL DEGRADATION AND FLAME RETARDANCY OF CALCIUM ALGINATE FIBERS

    Institute of Scientific and Technical Information of China (English)

    于建; 夏延致

    2009-01-01

    Calcium alginate fibers were prepared by wet spinning of sodium alginate into a coagulating bath containing calcium chloride.The thermal degradation and flame retardancy of calcium alginate fibers were investigated with thermal gravimetry(TG),X-ray diffraction(XRD),limiting oxygen index(LOI) and cone calorimeter(CONE).The results show that calcium alginate fibers are inherently flame retardant with a LOI value of 34,and the heat release rate(HRR),total heat release(THR),CO and CO_2 concentrations during ...

  20. Biosorption of americium by alginate beads

    International Nuclear Information System (INIS)

    Borba, Tania Regina de; Marumo, Julio Takehiro; Goes, Marcos Maciel de; Ferreira, Rafael Vicente de Padua; Sakata, Solange Kazumi

    2009-01-01

    The use of biotechnology to remove heavy metals from wastes plays great potential in treatment of radioactive wastes and therefore the aim of this study was to evaluate the biosorption of americium by alginate beads. Biosorption has been defined as the property of certain biomolecules to bind and remove selected ions or other molecules from aqueous solutions. The calcium alginate beads as biosorbent were prepared and analyzed for americium uptaking. The experiments were performed in different solution activity concentrations, pH and exposure time. The results suggest that biosorption process is more efficient at pH 4 and for 75, 150, 300 Bq/mL and 120 minutes were necessary to remove almost 100% of the americium-241 from the solution. (author)

  1. Microencapsulation of probiotics using sodium alginate

    Directory of Open Access Journals (Sweden)

    Mariana de Araújo Etchepare

    2015-07-01

    Full Text Available The consumption of probiotics is constantly growing due to the numerous benefits conferred on the health of consumers. In this context, Microencapsulation is a technology that favors the viability of probiotic cultures in food products, mainly by the properties of protection against adverse environmental conditions and controlled release. Currently there are different procedures for microencapsulation using polymers of various types of natural and synthetic origin. The use of sodium alginate polymers is one of the largest potential application in the encapsulation of probiotics because of their versatility, biocompatibility and toxicity exemption. The aim of this review is to present viable encapsulation techniques of probiotics with alginate, emphasizing the internal ionic gelation and external ionic gelation, with the possibility of applying, as well as promising for improving these techniques.

  2. 21 CFR 184.1133 - Ammonium alginate.

    Science.gov (United States)

    2010-04-01

    ...: Category of food Maximum level of use in food (as served) (percent) Functional use Confections, frostings... chapter 0.4 Do. Sweet sauces, § 170.3(n)(43) of this chapter 0.5 Do. All other food categories 0.1... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Ammonium alginate. 184.1133 Section 184.1133 Food...

  3. 21 CFR 184.1724 - Sodium alginate.

    Science.gov (United States)

    2010-04-01

    ....1(b)(2), the ingredient is used in food only within the following specific limitations: Category of...; texturizer, § 170.3(o)(32) of this chapter. All other food categories 1.0 Emulsifier, § 170.3(o)(8) of this... 21 Food and Drugs 3 2010-04-01 2009-04-01 true Sodium alginate. 184.1724 Section 184.1724 Food and...

  4. Alginate-Collagen Fibril Composite Hydrogel

    Directory of Open Access Journals (Sweden)

    Mahmoud Baniasadi

    2015-02-01

    Full Text Available We report on the synthesis and the mechanical characterization of an alginate-collagen fibril composite hydrogel. Native type I collagen fibrils were used to synthesize the fibrous composite hydrogel. We characterized the mechanical properties of the fabricated fibrous hydrogel using tensile testing; rheometry and atomic force microscope (AFM-based nanoindentation experiments. The results show that addition of type I collagen fibrils improves the rheological and indentation properties of the hydrogel.

  5. Inhibition of food-related bacteria by antibacterial substances produced by Pseudomonas sp. strains isolated from pasteurized milk

    Directory of Open Access Journals (Sweden)

    Ana Beatriz Ferreira Rangel

    2013-12-01

    Full Text Available In this work, the production of antimicrobial substances by strains of Pseudomonas sp. isolated from pasteurized milk and their potential action against food-related bacteria were investigated. Samples of pasteurized milk were purchased from arbitrarily chosen commercial establishments in the city of Rio de Janeiro, Brazil. Of the four samples analyzed, three presented several typical colonies of Pseudomonas. About 100 colonies were chosen and subjected to biochemical tests for confirmation of their identity. Eighteen strains of the Pseudomonas genus were identified and submitted to tests for the production of antimicrobial substances. Twelve strains (66.7% were identified as Pseudomonas fluorescens, four (22.2% as P. aeruginosa, one (5.5% as P. mendocina and one (5.5% as P. pseudoalcaligenes. Only two P. fluorescens strains were unable to produce any antimicrobial substance against any of the indicator strains tested. Most of the strains presented a broad spectrum of action, inhibiting reference and food-related strains such as Proteus vulgaris, Proteus mirabilis, Hafnia alvei, Yersinia enterocolitica, Escherichia coli and Salmonella typhi. Five antimicrobial substance-producing strains, which presented the broadest spectrum of action, were also tested against Staphylococcus aureus reference strains and 26 Staphylococcus sp. strains isolated from foods, some of which were resistant to antibiotics. The producer strains 8.1 and 8.3, both P. aeruginosa, were able to inhibit all the staphylococcal strains tested. The antimicrobial substances produced by strains 8.1 and 8.3 did not seem to be typical bacteriocins, since they were resistant to the three proteolytic enzymes tested. Experiments involving the characterization of these substances are being carried out in order to evaluate their biotechnological application.

  6. Glyphosate-Induced Specific and Widespread Perturbations in the Metabolome of Soil Pseudomonas Species

    Directory of Open Access Journals (Sweden)

    Ludmilla Aristilde

    2017-06-01

    Full Text Available Previous studies have reported adverse effects of glyphosate on crop-beneficial soil bacterial species, including several soil Pseudomonas species. Of particular interest is the elucidation of the metabolic consequences of glyphosate toxicity in these species. Here we investigated the growth and metabolic responses of soil Pseudomonas species grown on succinate, a common root exudate, and glyphosate at different concentrations. We conducted our experiments with one agricultural soil isolate, P. fluorescens RA12, and three model species, P. putida KT2440, P. putida S12, and P. protegens Pf-5. Our results demonstrated both species- and strain-dependent growth responses to glyphosate. Following exposure to a range of glyphosate concentrations (up to 5 mM, the growth rate of both P. protegens Pf-5 and P. fluorescens RA12 remained unchanged whereas the two P. putida strains exhibited from 0 to 100% growth inhibition. We employed a 13C-assisted metabolomics approach using liquid chromatography-mass spectrometry to monitor disruptions in metabolic homeostasis and fluxes. Profiling of the whole-cell metabolome captured deviations in metabolite levels involved in the tricarboxylic acid cycle, ribonucleotide biosynthesis, and protein biosynthesis. Altered metabolite levels specifically in the biosynthetic pathway of aromatic amino acids (AAs, the target of toxicity for glyphosate in plants, implied the same toxicity target in the soil bacterium. Kinetic flux experiments with 13C-labeled succinate revealed that biosynthetic fluxes of the aromatic AAs were not inhibited in P. fluorescens Pf-5 in the presence of low and high glyphosate doses but these fluxes were inhibited by up to 60% in P. putida KT2440, even at sub-lethal glyphosate exposure. Notably, the greatest inhibition was found for the aromatic AA tryptophan, an important precursor to secondary metabolites. When the growth medium was supplemented with aromatic AAs, P. putida S12 exposed to a lethal

  7. Effect of organic management of soils on suppressiveness to Gaeumannomyces graminis var. tritici and its antagonist, Pseudomonas fluorescens

    NARCIS (Netherlands)

    Hiddink, G.A.; Bruggen, van A.H.C.; Termorshuizen, A.J.; Raaijmakers, J.M.; Semenov, A.V.

    2005-01-01

    Organic management of soils is generally considered to reduce the incidence and severity of plant diseases caused by soil-borne pathogens. In this study, take-all severity on roots of barley and wheat, caused by Gaeumannomyces graminis var. tritici, was significantly lower in organically-managed

  8. No apparent costs for facultative antibiotic production by the soil bacterium Pseudomonas fluorescens Pf0-1

    NARCIS (Netherlands)

    Garbeva, P.V.; Tyc, O.; Remus-Emsermann, M.N.P.; Van der Wal, A.; Vos, M.; Silby, M.W.; De Boer, W.

    2011-01-01

    Background: Many soil-inhabiting bacteria are known to produce secondary metabolites that can suppress microorganisms competing for the same resources. The production of antimicrobial compounds is expected to incur fitness costs for the producing bacteria. Such costs form the basis for models on the

  9. Competition for nitrate and glucose between Pseudomonas fluorescens and Bacillus licheniformis under continuous or fluctuating anoxic conditions

    NARCIS (Netherlands)

    Nijburg, J.W.; Gerards, S.; Laanbroek, H.J.

    1998-01-01

    The dissimilatory nitrate-reducing bacterial community in the rhizosphere of aerenchymatous plant species such as Glyceria maxima, consists of oxidative. denitrifying and fermentative nitrate-ammonifying bacteria. To study the respective ecological niches of both types of nitrate-reducing bacteria,

  10. Competition for nitrate and glucose between Pseudomonas fluorescens and Bacillus licheniformis under continuous or fluctuating anoxic conditions

    NARCIS (Netherlands)

    Nijburg, J.W.; Gerards, S.; Laanbroek, H.J.

    1998-01-01

    The dissimilatory nitrate-reducing bacterial community in the rhizosphere of aerenchymatous plant species such as Glyceria maxima, consists of oxidative, denitrifying and fermentative nitrate-ammonifying bacteria. To study the respective ecological niches of both types of nitrate-reducing

  11. Polychlorinated biphenyl rhizoremediation by Pseudomonas fluorescens F113 derivatives, using a Sinorhizobium meliloti nod system to drive bph gene expression

    Czech Academy of Sciences Publication Activity Database

    Villacieros, M.; Whelan, C.; Macková, Martina; Molgaard, J.; Sánchez-Contreras, M.; Lloret, J.; Cárcer de, D.; Oruezábal, R. I.; Bolanos, L.; Macek, Tomáš; Karlson, U.; Dowling, D. N.; Martín, M.; Rivilla, R.

    2005-01-01

    Roč. 71, č. 5 (2005), 2687-2694 ISSN 0099-2240 Grant - others:5thFW(XE) QLK3-CT-2001-00101; European Commission(XE) BIO4-CT97-2227 Institutional research plan: CEZ:AV0Z4055905 Keywords : polychlorinated biphenyls * rhizoremediation Subject RIV: CC - Organic Chemistry Impact factor: 3.818, year: 2005

  12. Pseudomonas fluorescens HK44: Lessons Lerned from a Model Whole-Cell Bioreporter with a Broad Application History

    Czech Academy of Sciences Publication Activity Database

    Trögl, J.; Chauhan, A.; Ripp, S.; Layton, A.C.; Kuncová, Gabriela; Sayler, G.S.

    2012-01-01

    Roč. 12, č. 2 (2012), s. 1544-1571 ISSN 1424-8220 R&D Projects: GA MŠk ME 892; GA MŠk ME 893 Grant - others:USDA(US) 2009-39210-20230 Institutional research plan: CEZ:AV0Z40720504 Keywords : bioluminiscence * bioreporter * biosensors Subject RIV: EI - Biotechnology ; Bionics Impact factor: 1.953, year: 2012

  13. Controlled release studies of calcium alginate hydrogels

    International Nuclear Information System (INIS)

    Rendevski, S.; Andonovski, A.; Mahmudi, N.

    2012-01-01

    Controlled release of substances in many cases may be achieved from calcium alginate hydrogels. In this research, the time dependence of the mass of released model substance bovine serum albumin (BSA) from calcium alginate spherical hydrogels of three different types (G/M ratio) have been investigated. The hydrogels were prepared with the drop-wise method of sodium alginate aqueous solutions with concentration of 0.02 g/cm 3 with 0.01 g/cm 3 BSA and a gelling water bath of chitosan in 0.2 M CH 3 COOH/0.4 M CH 3 COONa with added 0.2 M CaCl 2 .The hydrogel structures were characterized by dynamic light scattering and scanning electron microscopy. The controlled release studies were conducted by UV-Vis spectrophotometry of the released medium with p H=7 at 37 °C. The results showed that the model of osmotic pumping is the dominant mechanism of the release. Also, large dependences of the release profile on the homogeneity of the hydrogels were found. (Author)

  14. Gentamicin in Pseudomonas aeruginosa

    African Journals Online (AJOL)

    infections by Ps. aeruginosa is contra-indicated. In our study only 2,3 % of the Ps. aeruginosa strains were resistant to gentamicin (MIC 25 Ilg/ml). In view of the synergy reported for combined gentamicin and carbeni- cillin therapy," a combination of these two drugs may be recommended in the treatment of all Pseudomonas.

  15. Uptake of Hydrocarbon by Pseudomonas fluorescens (P1) and Pseudomonas putida (K1) Strains in the Presence of Surfactants: A Cell Surface Modification

    OpenAIRE

    Kaczorek, Ewa; Olszanowski, Andrzej

    2010-01-01

    The objective of this research was the evaluation of the effects of exogenous added surfactants on hydrocarbon biodegradation and on cell surface properties. Crude oil hydrocarbons are often difficult to remove from the environment because of their insolubility in water. The addition of surfactants enhances the removal of hydrocarbons by raising the solubility of these compounds. These surfactants cause them to become more vulnerable to degradation, thereby facilitating transportation across ...

  16. Influence of putative exopolysaccharide genes on Pseudomonas putida KT2440 biofilm stability

    DEFF Research Database (Denmark)

    Nilsson, Martin; Chiang, Wen-Chi; Fazli, Mustafa

    2011-01-01

    We report a study of the role of putative exopolysaccharide gene clusters in the formation and stability of Pseudomonas putida KT2440 biofilm. Two novel putative exopolysaccharide gene clusters, pea and peb, were identified, and evidence is provided that they encode products that stabilize P....... putida KT2440 biofilm. The gene clusters alg and bcs, which code for proteins mediating alginate and cellulose biosynthesis, were found to play minor roles in P. putida KT2440 biofilm formation and stability under the conditions tested. A P. putida KT2440 derivative devoid of any identifiable...

  17. Pseudomonas cichorii as the causal agent of midrib rot, an emerging disease of greenhouse-grown butterhead lettuce in Flanders.

    Science.gov (United States)

    Cottyn, Bart; Heylen, Kim; Heyrman, Jeroen; Vanhouteghem, Katrien; Pauwelyn, Ellen; Bleyaert, Peter; Van Vaerenbergh, Johan; Höfte, Monica; De Vos, Paul; Maes, Martine

    2009-05-01

    Bacterial midrib rot of greenhouse-grown butterhead lettuce (Lactuca sativa L. var. capitata) is an emerging disease in Flanders (Belgium) and fluorescent pseudomonads are suspected to play an important role in the disease. Isolations from infected lettuces, collected from 14 commercial greenhouses in Flanders, yielded 149 isolates that were characterized polyphasically, which included morphological characteristics, pigmentation, pathogenicity tests by both injection and spraying of lettuce, LOPAT characteristics, FAME analysis, BOX-PCR fingerprinting, 16S rRNA and rpoB gene sequencing, as well as DNA-DNA hybridization. Ninety-eight isolates (66%) exhibited a fluorescent pigmentation and were associated with the genus Pseudomonas. Fifty-five of them induced an HR+ (hypersensitive reaction in tobacco leaves) response. The other 43 fluorescent isolates were most probably saprophytic bacteria and about half of them were able to cause rot on potato tuber slices. BOX-PCR genomic fingerprinting was used to assess the genetic diversity of the Pseudomonas midrib rot isolates. The delineated BOX-PCR patterns matched quite well with Pseudomonas morphotypes defined on the basis of colony appearance and variation in fluorescent pigmentation. 16S rRNA and rpoB gene sequence analyses allowed most of the fluorescent isolates to be allocated to Pseudomonas, and they belonged to either the Pseudomonas fluorescens group, Pseudomonas putida group, or the Pseudomonas cichorii/syringae group. In particular, the isolates allocated to this latter group constituted the vast majority of HR+ isolates and were identified as P. cichorii by DNA-DNA hybridization. They were demonstrated by spray-inoculation tests on greenhouse-grown lettuce to induce the midrib rot disease and could be re-isolated from lesions of inoculated plants. Four HR+ non-fluorescent isolates associated with one sample that showed an atypical midrib rot were identified as Dickeya sp.

  18. A Controlled Drug-Delivery Experiment Using Alginate Beads

    Science.gov (United States)

    Farrell, Stephanie; Vernengo, Jennifer

    2012-01-01

    This paper describes a simple, cost-effective experiment which introduces students to drug delivery and modeling using alginate beads. Students produce calcium alginate beads loaded with drug and measure the rate of release from the beads for systems having different stir rates, geometries, extents of cross-linking, and drug molecular weight.…

  19. The complete genome sequence of the plant growth-promoting bacterium Pseudomonas sp. UW4.

    Directory of Open Access Journals (Sweden)

    Jin Duan

    Full Text Available The plant growth-promoting bacterium (PGPB Pseudomonas sp. UW4, previously isolated from the rhizosphere of common reeds growing on the campus of the University of Waterloo, promotes plant growth in the presence of different environmental stresses, such as flooding, high concentrations of salt, cold, heavy metals, drought and phytopathogens. In this work, the genome sequence of UW4 was obtained by pyrosequencing and the gaps between the contigs were closed by directed PCR. The P. sp. UW4 genome contains a single circular chromosome that is 6,183,388 bp with a 60.05% G+C content. The bacterial genome contains 5,423 predicted protein-coding sequences that occupy 87.2% of the genome. Nineteen genomic islands (GIs were predicted and thirty one complete putative insertion sequences were identified. Genes potentially involved in plant growth promotion such as indole-3-acetic acid (IAA biosynthesis, trehalose production, siderophore production, acetoin synthesis, and phosphate solubilization were determined. Moreover, genes that contribute to the environmental fitness of UW4 were also observed including genes responsible for heavy metal resistance such as nickel, copper, cadmium, zinc, molybdate, cobalt, arsenate, and chromate. Whole-genome comparison with other completely sequenced Pseudomonas strains and phylogeny of four concatenated "housekeeping" genes (16S rRNA, gyrB, rpoB and rpoD of 128 Pseudomonas strains revealed that UW4 belongs to the fluorescens group, jessenii subgroup.

  20. The Complete Genome Sequence of the Plant Growth-Promoting Bacterium Pseudomonas sp. UW4

    Science.gov (United States)

    Duan, Jin; Jiang, Wei; Cheng, Zhenyu; Heikkila, John J.; Glick, Bernard R.

    2013-01-01

    The plant growth-promoting bacterium (PGPB) Pseudomonas sp. UW4, previously isolated from the rhizosphere of common reeds growing on the campus of the University of Waterloo, promotes plant growth in the presence of different environmental stresses, such as flooding, high concentrations of salt, cold, heavy metals, drought and phytopathogens. In this work, the genome sequence of UW4 was obtained by pyrosequencing and the gaps between the contigs were closed by directed PCR. The P. sp. UW4 genome contains a single circular chromosome that is 6,183,388 bp with a 60.05% G+C content. The bacterial genome contains 5,423 predicted protein-coding sequences that occupy 87.2% of the genome. Nineteen genomic islands (GIs) were predicted and thirty one complete putative insertion sequences were identified. Genes potentially involved in plant growth promotion such as indole-3-acetic acid (IAA) biosynthesis, trehalose production, siderophore production, acetoin synthesis, and phosphate solubilization were determined. Moreover, genes that contribute to the environmental fitness of UW4 were also observed including genes responsible for heavy metal resistance such as nickel, copper, cadmium, zinc, molybdate, cobalt, arsenate, and chromate. Whole-genome comparison with other completely sequenced Pseudomonas strains and phylogeny of four concatenated “housekeeping” genes (16S rRNA, gyrB, rpoB and rpoD) of 128 Pseudomonas strains revealed that UW4 belongs to the fluorescens group, jessenii subgroup. PMID:23516524

  1. Arsenic-contaminated soils. Genetically modified Pseudomonas spp. and their arsenic-phytoremediation potential

    Energy Technology Data Exchange (ETDEWEB)

    Sizova, O.I.; Kochetkov, V.V.; Validov, S.Z.; Boronin, A.M. [Inst. of Biochemistry and Physiology of Microorganisms, Russian Academy of Sciences, Moscow (Russian Federation); Kosterin, P.V.; Lyubun, Y.V. [Inst. of Biochemistry and Physiology of Plants and Microorganisms, Russian Academy of Sciences, Saratov (Russian Federation)

    2002-07-01

    Sorghum was inoculated with Pseudomonas bacteria, including strains harboring an As-resistance plasmid, pBS3031, to enhance As-extraction by the plants. Pseudomonas strains (P. fluorescens 38a, P. putida 53a, and P. aureofaciens BS1393) were chosen because they are antagonistic to a wide range of phytopathogenic fungi and bacteria, and they can stimulate plant growth. The resistance of natural rhizospheric pseudomonads to sodium arsenite was assessed. Genetically modified Pseudomonas strains resistant to As(III)/As(V) were obtained via conjugation or transformation. The effects of the strains on the growth of sorghum on sodium-arsenite-containing soils were assessed. The conclusions from this study are: (1) It is possible to increase the survivability of sorghum growing in sodium-arsenite-containing soil by using rhizosphere pseudomonads. (2) The presence of pBS3031 offers the strains a certain selective advantage in arsenite-contaminated soil. (3) The presence of pBS3031 impairs plant growth, due to the As-resistance mechanism determined by this plasmid: the transformation of the less toxic arsenate into the more toxic, plant-root-available arsenite by arsenate reductase and the active removal of arsenite from bacterial cells. (4) Such a mechanism makes it possible to develop a bacteria-assisted phytoremediation technology for the cleanup of As-contaminated soils and is the only possible way of removing the soil-sorbed arsenates from the environment. (orig.)

  2. Applications of Alginate-Based Bioinks in 3D Bioprinting

    Science.gov (United States)

    Axpe, Eneko; Oyen, Michelle L.

    2016-01-01

    Three-dimensional (3D) bioprinting is on the cusp of permitting the direct fabrication of artificial living tissue. Multicellular building blocks (bioinks) are dispensed layer by layer and scaled for the target construct. However, only a few materials are able to fulfill the considerable requirements for suitable bioink formulation, a critical component of efficient 3D bioprinting. Alginate, a naturally occurring polysaccharide, is clearly the most commonly employed material in current bioinks. Here, we discuss the benefits and disadvantages of the use of alginate in 3D bioprinting by summarizing the most recent studies that used alginate for printing vascular tissue, bone and cartilage. In addition, other breakthroughs in the use of alginate in bioprinting are discussed, including strategies to improve its structural and degradation characteristics. In this review, we organize the available literature in order to inspire and accelerate novel alginate-based bioink formulations with enhanced properties for future applications in basic research, drug screening and regenerative medicine. PMID:27898010

  3. Applications of Alginate-Based Bioinks in 3D Bioprinting.

    Science.gov (United States)

    Axpe, Eneko; Oyen, Michelle L

    2016-11-25

    Three-dimensional (3D) bioprinting is on the cusp of permitting the direct fabrication of artificial living tissue. Multicellular building blocks (bioinks) are dispensed layer by layer and scaled for the target construct. However, only a few materials are able to fulfill the considerable requirements for suitable bioink formulation, a critical component of efficient 3D bioprinting. Alginate, a naturally occurring polysaccharide, is clearly the most commonly employed material in current bioinks. Here, we discuss the benefits and disadvantages of the use of alginate in 3D bioprinting by summarizing the most recent studies that used alginate for printing vascular tissue, bone and cartilage. In addition, other breakthroughs in the use of alginate in bioprinting are discussed, including strategies to improve its structural and degradation characteristics. In this review, we organize the available literature in order to inspire and accelerate novel alginate-based bioink formulations with enhanced properties for future applications in basic research, drug screening and regenerative medicine.

  4. Evolutionary Plasticity of AmrZ Regulation in Pseudomonas

    Science.gov (United States)

    Dougherty, Kevin; Diaz, Beatriz; Murillo, Rachel

    2018-01-01

    ABSTRACT amrZ encodes a master regulator protein conserved across pseudomonads, which can be either a positive or negative regulator of swimming motility depending on the species examined. To better understand plasticity in the regulatory function of AmrZ, we characterized the mode of regulation for this protein for two different motility-related phenotypes in Pseudomonas stutzeri. As in Pseudomonas syringae, AmrZ functions as a positive regulator of swimming motility within P. stutzeri, which suggests that the functions of this protein with regard to swimming motility have switched at least twice across pseudomonads. Shifts in mode of regulation cannot be explained by changes in AmrZ sequence alone. We further show that AmrZ acts as a positive regulator of colony spreading within this strain and that this regulation is at least partially independent of swimming motility. Closer investigation of mechanistic shifts in dual-function regulators like AmrZ could provide unique insights into how transcriptional pathways are rewired between closely related species. IMPORTANCE Microbes often display finely tuned patterns of gene regulation across different environments, with major regulatory changes controlled by a small group of “master” regulators within each cell. AmrZ is a master regulator of gene expression across pseudomonads and can be either a positive or negative regulator for a variety of pathways depending on the strain and genomic context. Here, we demonstrate that the phenotypic outcomes of regulation of swimming motility by AmrZ have switched at least twice independently in pseudomonads, so that AmrZ promotes increased swimming motility in P. stutzeri and P. syringae but represses this phenotype in Pseudomonas fluorescens and Pseudomonas aeruginosa. Since examples of switches in regulatory mode are relatively rare, further investigation into the mechanisms underlying shifts in regulator function for AmrZ could provide unique insights into the

  5. [Nah-plasmids of IncP-9 group from natural strains of Pseudomonas].

    Science.gov (United States)

    Levchuk, A A; Bulyga, I M; Izmalkova, T Iu; Sevast'ianovich, Ia R; Kosheleva, I A; Thomas, C M; Titok, M A

    2006-01-01

    Use of polymerase chain reaction helped to establish that the most frequent among naphthalene utilizing bacteria, isolated on the territory of Belarus, are Nah-plasmids of IncP-9 incompatibility group and those with indefinite systematic belonging. With the help of classical test of incompatibility, restriction and sequence analyses three new subgroups within the IncP-9 group were discovered (zeta, eta and IncP-9-like replicons). Conducting of restriction analysis for amplification products of nahG and nahAc genes allowed us to reveal, in addition to known sequences of stated determinants, two new types of nahG gene. Restriction analysis performed on amplification products of 16S RNA genes (ARDRA method) showed that native hosts of Nah-plasmids of IncP-9 group are not only fluorescent bacteria from genus Pseudomonas (P. fluorescens, P. putida, P. aeruginosa, P. species), but also non-fluorescent bacteria with indefinite specific belonging.

  6. Dimensional changes of alginate dental impression materials.

    Science.gov (United States)

    Nallamuthu, N; Braden, M; Patel, M P

    2006-12-01

    The weight loss and corresponding dimensional changes of two dental alginate impression materials have been studied. The weight loss kinetics indicate this to be a diffusion controlled process, but with a boundary condition at the surface of the concentration decreasing exponentially with time. This is in marked contrast to most desorption processes, where the surface concentration becomes instantaneously zero. The appropriate theory has been developed for an exponential boundary condition, and its predictions compared with experimental data; the agreement was satisfactory. The diffusion coefficients for two thicknesses of the same material were not identical as predicted by theory; the possible reasons for this are discussed.

  7. A novel enterocin T1 with anti-Pseudomonas activity produced by Enterococcus faecium T1 from Chinese Tibet cheese.

    Science.gov (United States)

    Liu, Hui; Zhang, Lanwei; Yi, Huaxi; Han, Xue; Gao, Wei; Chi, Chunliang; Song, Wei; Li, Haiying; Liu, Chunguang

    2016-02-01

    An enterocin-producing Enterococcus faecium T1 was isolated from Chinese Tibet cheese. The enterocin was purified by SP-Sepharose and reversed phase HPLC. It was identified as unique from other reported bacteriocins based on molecular weight (4629 Da) and amino acid compositions; therefore it was subsequently named enterocin T1. Enterocin T1 was stable at 80-100 °C and over a wide pH range, pH 3.0-10.0. Protease sensitivity was observed to trypsin, pepsin, papain, proteinase K, and pronase E. Importantly, enterocin T1 was observed to inhibit the growth of numerous Gram-negative and Gram-positive bacteria including Pseudomonas putida, Pseudomonas aeruginosa, Pseudomonas fluorescens, Escherichia coli, Salmonella typhimurium, Shigella flexneri, Shigella sonnei, Staphylococcus aureus, Listeria monocytogenes. Take together, these results suggest that enterocin T1 is a novel bacteriocin with the potential to be used as a bio-preservative to control Pseudomonas spp. in food.

  8. Design and evaluate alginate nanoparticles as a protein delivery system

    Directory of Open Access Journals (Sweden)

    Saraei, F.

    2013-12-01

    Full Text Available In recent years, encapsulation of drugs and antigens in hydrogels, specifically in calcium alginate particles, is an interesting and practical technique that was developed widespread. It is well known that alginate solution, under proper conditions, can form suitable nanoparticles as a promising carrier system, for vaccine delivery. The aim of this study was to synthesis alginate nanoparticles as protein carrier and to evaluate the influence of various factors on nanoparticles properties. Alginate nanoparticles were prepared by ionic gelation method. Briefly, various concentrations of CaCl2 were added to different concentrations of sodium alginate dropwisly by homogenizing magnetically at 1300 rpm. The effects of homogenization time and (- rate were investigated on nanoparticle feature. Nanoparticles were characterized for their morphology and size distribution. Evaluation of loading capacity and loading efficiency of nanoparticles were performed by using various concentration of BSA. The concentration of 0.3%w/v sodium alginate and 0.1%w/v CaCl2 solution, homogenization time 45 min and homogenization rate 1300 rpm were observed as suitable condition - to prepare optimized nanoparticles. It can be concluded that the properties of nanoparticles are strongly dependent on the physicochemical conditions. The optimum concentrations of alginate and CaCl2and appropriate condition led to forming desirable nanoparticles that can be used as carrier for drug and vaccine delivery.

  9. Alginate based scaffolds for bone tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Valente, J.F.A.; Valente, T.A.M. [CICS-UBI - Centro de Investigacao em Ciencias da Saude, Faculdade de Ciencias da Saude, Universidade da Beira Interior, Covilha (Portugal); Alves, P.; Ferreira, P. [CIEPQPF, Departamento de Engenharia Quimica, Universidade de Coimbra, Polo II, Pinhal de Marrocos, 3030-290 Coimbra (Portugal); Silva, A. [Centro de Ciencia e Tecnologia Aeroespaciais, Universidade da Beira Interior, Covilha (Portugal); Correia, I.J., E-mail: icorreia@ubi.pt [CICS-UBI - Centro de Investigacao em Ciencias da Saude, Faculdade de Ciencias da Saude, Universidade da Beira Interior, Covilha (Portugal)

    2012-12-01

    The design and production of scaffolds for bone tissue regeneration is yet unable to completely reproduce the native bone properties. In the present study new alginate microparticle and microfiber aggregated scaffolds were produced to be applied in this area of regenerative medicine. The scaffolds' mechanical properties were characterized by thermo mechanical assays. Their morphological characteristics were evaluated by isothermal nitrogen adsorption and scanning electron microscopy. The density of both types of scaffolds was determined by helium pycnometry and mercury intrusion porosimetry. Furthermore, scaffolds' cytotoxic profiles were evaluated in vitro by seeding human osteoblast cells in their presence. The results obtained showed that scaffolds have good mechanical and morphological properties compatible with their application as bone substitutes. Moreover, scaffold's biocompatibility was confirmed by the observation of cell adhesion and proliferation after 5 days of being seeded in their presence and by non-radioactive assays. - Highlights: Black-Right-Pointing-Pointer Design and production of scaffolds for bone tissue regeneration. Black-Right-Pointing-Pointer Microparticle and microfiber alginate scaffolds were produced through a particle aggregation technique; Black-Right-Pointing-Pointer Scaffolds' mechanically and biologically properties were characterized through in vitro studies;.

  10. Alginate production affects Pseudomonas aeruginosa biofilm development and architecture, but is not essential for biofilm formation

    DEFF Research Database (Denmark)

    Stapper, A.P.; Narasimhan, G.; Oman, D.E.

    2004-01-01

    of their biofilm formation using confocal laser scanning microscopy. Biofilm Image Processing (BIP) and Community Statistics (COMSTAT) software programs were used to provide quantitative measurements of the two-dimensional biofilm images. All three strains formed distinguishable biofilm architectures, indicating...

  11. Antimicrobial activity of gallic acid against food-related Pseudomonas strains and its use as biocontrol tool to improve the shelf life of fresh black truffles.

    Science.gov (United States)

    Sorrentino, Elena; Succi, Mariantonietta; Tipaldi, Luca; Pannella, Gianfranco; Maiuro, Lucia; Sturchio, Marina; Coppola, Raffaele; Tremonte, Patrizio

    2018-02-02

    Refrigeration alone or in combination with other technologies represents the main tool used in the last decades to preserve the freshness of black truffles. This is principally due to the delicateness and vulnerability of this edible hypogeous fungus, so that other invasive preservation practices cannot be adopted. However, the proliferation of some microbial species during the cold storage still represents an unsolved problem. Pseudomonads are among the main spoiler bacteria responsible for the deterioration of refrigerated black truffles. Their growth ability at low temperatures requires the use of additional hurdles to prolong the shelf-life of truffles without altering their major features. The use of natural compounds may represent an alternative system for the biocontrol of this kind of product. Specifically, gallic acid (GA) is a phenolic acid naturally present in different foods, whose effectiveness was in vitro demonstrated against Pseudomonas spp. In our study, we reported the antimicrobial activity expressed by GA not only in vitro, using as target bacteria Pseudomonas putida DSMZ 291 T , P. fluorescens DSMZ 50090 T , P. fragi DSMZ 3456 T and Pseudomonas spp. P30-4, previously isolated from black truffles, but also in situ on fresh black truffles stored at 4°C for 28days. Our results showed Minimum Inhibitory Concentrations (MIC) of 2.5mg/mL GA for all tested strains, except for P. fluorescens DSMZ 50090 T , having a MIC corresponding to 5mg/mL GA. The Minimum Bactericidal Concentration (MBC) was 10mg/mL for all strains. The analysis of kinetic parameters showed that the survival declined passing from 2.5 to 10mg/mL GA concentrations, with P. fluorescens confirmed to be the most resistant strain. Moreover, images obtained from Scanning Electron Microscopy revealed that Pseudomonas cells were strongly injured by the treatment with GA at 2.5mg/mL concentration, displaying visible pores on the cellular surfaces, absence of flagella and lysis with loss of

  12. Pseudomonas aeruginosa biofilm aggravates skin inflammatory response in BALB/c mice in a novel chronic wound model

    DEFF Research Database (Denmark)

    Trøstrup, Hannah; Thomsen, Kim; Christophersen, Lars J

    2013-01-01

    model in C3H/HeN and BALB/c mice. The chronic wound was established by an injection of seaweed alginate-embedded P. aeruginosa PAO1 beneath a third-degree thermal lesion providing full thickness skin necrosis, as in human chronic wounds. Cultures revealed growth of PA, and both alginate with or without......Chronic wounds are presumed to persist in the inflammatory state, preventing healing. Emerging evidence indicates a clinical impact of bacterial biofilms in soft tissues, including Pseudomonas aeruginosa (PA) biofilms. To further investigate this, we developed a chronic PA biofilm wound infection...... bacteria organized in clusters, resembling biofilms, and inflammation located adjacent to the PA. The chronic wound infection showed a higher number of PAO1 in the BALB/c mice at day 4 after infection as compared to C3H/HeN mice (p

  13. The influence of storage duration on the setting time of type 1 alginate impression material

    Science.gov (United States)

    Rahmadina, A.; Triaminingsih, S.; Irawan, B.

    2017-08-01

    Alginate is one of the most commonly used dental impression materials; however, its setting time is subject to change depending on storage conditions and duration. This creates problems because consumer carelessness can affect alginate shelf life and quality. In the present study, the setting times of two groups of type I alginate with different expiry dates was tested. The first group consisted of 11 alginate specimens that had not yet passed the expiry date, and the second group consisted of alginates that had passed the expiry date. The alginate powder was mixed with distilled water, poured into a metal ring, and tested with a polished rod of poly-methyl methacrylate. Statistical analysis showed a significant difference (p<0.05) between the setting times of the alginate that had not passed the expiry date (157 ± 3 seconds) and alginate that had passed the expiry date (144 ± 2 seconds). These findings indicate that storage duration can affect alginate setting time.

  14. Adsorption studies of phosphate ions on alginate- calcium ...

    African Journals Online (AJOL)

    user

    osmosis, electro dialysis, phytoremediation and phyto- extraction, ion-exchange .... occurred between functional groups of alginate and host material, that is, calcium .... metal ions using wheat based biosorbent; a review of the recent literature.

  15. Ca alginate as scaffold for iron oxide nanoparticles synthesis

    Directory of Open Access Journals (Sweden)

    P. V. Finotelli

    2008-12-01

    Full Text Available Recently, nanotechnology has developed to a stage that makes it possible to process magnetic nanoparticles for the site-specific delivery of drugs. To this end, it has been proposed as biomaterial for drug delivery system in which the drug release rates would be activated by a magnetic external stimuli. Alginate has been used extensively in the food, pharmaceutical and biomedical industries for their gel forming properties in the presence of multivalent cations. In this study, we produced iron oxide nanoparticles by coprecipitation of Fe(III and Fe(II. The nanoparticles were entrapped in Ca alginate beads before and after alginate gelation. XRD analysis showed that particles should be associated to magnetite or maghemite with crystal size of 9.5 and 4.3 nm, respectively. Studies using Mössbauer spectroscopy corroborate the superparamagnetic behavior. The combination of magnetic properties and the biocompatibility of alginate suggest that this biomaterial may be used as biomimetic system.

  16. The Alginate Demonstration: Polymers, Food Science, and Ion Exchange

    Science.gov (United States)

    Waldman, Amy Sue; Schechinger, Linda; Govindarajoo, Geeta; Nowick, James S.; Pignolet, Louis H.

    1998-11-01

    We have recently devised a polymer demonstration involving the crosslinking and decrosslinking of alginate, a polysaccharide isolated from seaweed. The polymer is composed of D-mannuronic acid and L-guluronic acid subunits and is a component of cell walls. It is commonly used as a thickener in foods such as ice cream and fruit-filled snacks. For the demonstration, a 2% solution of sodium alginate is poured into a 1% solution of calcium chloride. Nontoxic calcium alginate "worms" form due to crosslinking of the polymer. Alternatively, the commercially available antacid Gaviscon can be used as a source of sodium alginate. The crosslinks can then be broken by shaking the worms in brine. The demonstration is a fine addition to any chemical educator's repertoire of polymer experiments.

  17. Alginate-Chitosan Particulate System for Sustained Release of ...

    African Journals Online (AJOL)

    Erah

    1School of Pharmacy and Health Sciences, International Medical University, Kuala Lumpur, Malaysia, 2Division of ... Both calcium alginate beads and the beads treated with chitosan failed to release ..... also found to fit the classical power law.

  18. A Phytoanticipin Derivative, Sodium Houttuyfonate, Induces in Vitro Synergistic Effects with Levofloxacin against Biofilm Formation by Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Jing Shao

    2012-09-01

    Full Text Available Antibiotic resistance has become the main deadly factor in infections, as bacteria can protect themselves by hiding in a self-constructed biofilm. Consequently, more attention is being paid to the search for “non-antibiotic drugs” to solve this problem. Phytoanticipins, the natural antibiotics from plants, could be a suitable alternative, but few works on this aspect have been reported. In this study, a preliminary study on the synergy between sodium houttuyfonate (SH and levofloxacin (LFX against the biofilm formation of Pseudomonas aeruginosa was performed. The minimal inhibitory concentrations (MIC of LFX and SH, anti-biofilm formation and synergistic effect on Pseudomonas aeruginosa, and quantification of alginate were determined by the microdilution method, crystal violet (CV assay, checkerboard method, and hydroxybiphenyl colorimetry. The biofilm morphology of Pseudomonas aeruginosa was observed by fluorescence microscope and scanning electric microscope (SEM. The results showed that: (i LFX and SH had an obvious synergistic effect against Pseudomonas aeruginosa with MIC values of 0.25 μg/mL and 128 μg/mL, respectively; (ii ½ × MIC SH combined with 2 × MIC LFX could suppress the biofilm formation of Pseudomonas aeruginosa effectively, with up to 73% inhibition; (iii the concentration of alginate decreased dramatically by a maximum of 92% after treatment with the combination of antibiotics; and (iv more dead cells by fluorescence microscope and more removal of extracellular polymeric structure (EPS by SEM were observed after the combined treatment of LFX and SH. Our experiments demonstrate the promising future of this potent antimicrobial agent against biofilm-associated infections.

  19. Preparation of alginate coated chitosan microparticles for vaccine delivery

    Directory of Open Access Journals (Sweden)

    Wei YuQuan

    2008-11-01

    Full Text Available Abstract Background Absorption of antigens onto chitosan microparticles via electrostatic interaction is a common and relatively mild process suitable for mucosal vaccine. In order to increase the stability of antigens and prevent an immediate desorption of antigens from chitosan carriers in gastrointestinal tract, coating onto BSA loaded chitosan microparticles with sodium alginate was performed by layer-by-layer technology to meet the requirement of mucosal vaccine. Results The prepared alginate coated BSA loaded chitosan microparticles had loading efficiency (LE of 60% and loading capacity (LC of 6% with mean diameter of about 1 μm. When the weight ratio of alginate/chitosan microparticles was greater than 2, the stable system could be obtained. The rapid charge inversion of BSA loaded chitosan microparticles (from +27 mv to -27.8 mv was observed during the coating procedure which indicated the presence of alginate layer on the chitosan microparticles surfaces. According to the results obtained by scanning electron microscopy (SEM, the core-shell structure of BSA loaded chitosan microparticles was observed. Meanwhile, in vitro release study indicated that the initial burst release of BSA from alginate coated chitosan microparticles was lower than that observed from uncoated chitosan microparticles (40% in 8 h vs. about 84% in 0.5 h. SDS-polyacrylamide gel electrophoresis (SDS-PAGE assay showed that alginate coating onto chitosan microparticles could effectively protect the BSA from degradation or hydrolysis in acidic condition for at least 2 h. The structural integrity of alginate modified chitosan microparticles incubated in PBS for 24 h was investigated by FTIR. Conclusion The prepared alginate coated chitosan microparticles, with mean diameter of about 1 μm, was suitable for oral mucosal vaccine. Moreover, alginate coating onto the surface of chitosan microparticles could modulate the release behavior of BSA from alginate coated chitosan

  20. Alginate prevention of internal irradiation with 90Sr

    International Nuclear Information System (INIS)

    Korzun, V.N.; Voronova, Yu.G.; Parats, A.N.; Podkorytova, A.V.; Rogal'skaya, L.A.; Saglo, V.I.; Skorikova, A.I.

    1992-01-01

    Recipes of foodstaffs (meat and vegetable preserves, bread, pastry, dairy products, etc.) containing sodium or calcium alginates in doses 0.5-3.0 g have been developed. Experiments with white rats have demonstrated that addition of such products to daily radions of these animals reduced 2-4-fold the accumulation of radioactive Sr taken daily with food for 30 days. Alginates and Crambe added to food preserve their ability to reduce the accumulation of radioactive Sr

  1. Study on biosorption of uranium by alginate immobilized saccharomyces cerevisiae

    International Nuclear Information System (INIS)

    Wang Baoe; Xu Weichang; Xie Shuibo; Guo Yangbin

    2005-01-01

    Saccharomyces cerevisiae has great capability of biosorption of uranium. The maxium uptake is 172.4 mg/g according to this study. To adapt to the application of the biomass in the field, the biosorption of uranium by cross-linked and alginate calcium immobilized Saccharomyces cerevisiae is studied. Results indicate the maxium uptake is 185.2 mg/g by formaldehyde cross-linked biomass, and it is 769.2 mg/g by alginate calcium immobilized biomass. (authors)

  2. Alginate: A Versatile Biomaterial to Encapsulate Isolated Ovarian Follicles.

    Science.gov (United States)

    Vanacker, Julie; Amorim, Christiani A

    2017-07-01

    In vitro culture of ovarian follicles isolated or enclosed in ovarian tissue fragments and grafting of isolated ovarian follicles represent a potential alternative to restore fertility in cancer patients who cannot undergo cryopreservation of embryos or oocytes or transplantation of frozen-thawed ovarian tissue. In this regard, respecting the three-dimensional (3D) architecture of isolated follicles is crucial to maintaining their proper follicular physiology. To this end, alginate hydrogel has been widely investigated using follicles from numerous animal species, yielding promising results. The goal of this review is therefore to provide an overview of alginate applications utilizing the biomaterial as a scaffold for 3D encapsulation of isolated ovarian follicles. Different methods of isolated follicle encapsulation in alginate are discussed in this review, as its use of 3D alginate culture systems as a tool for in vitro follicle analysis. Possible improvements of this matrix, namely modification with arginine-glycine-aspartic acid peptide or combination with fibrin, are also summarized. Encouraging results have been obtained in different animal models, and particularly with isolated follicles encapsulated in alginate matrices and grafted to mice. This summary is designed to guide the reader towards development of next-generation alginate scaffolds, with enhanced properties for follicle encapsulation.

  3. Comparative genomics of plant-associated Pseudomonas spp.: insights into diversity and inheritance of traits involved in multitrophic interactions.

    Directory of Open Access Journals (Sweden)

    Joyce E Loper

    2012-07-01

    Full Text Available We provide here a comparative genome analysis of ten strains within the Pseudomonas fluorescens group including seven new genomic sequences. These strains exhibit a diverse spectrum of traits involved in biological control and other multitrophic interactions with plants, microbes, and insects. Multilocus sequence analysis placed the strains in three sub-clades, which was reinforced by high levels of synteny, size of core genomes, and relatedness of orthologous genes between strains within a sub-clade. The heterogeneity of the P. fluorescens group was reflected in the large size of its pan-genome, which makes up approximately 54% of the pan-genome of the genus as a whole, and a core genome representing only 45-52% of the genome of any individual strain. We discovered genes for traits that were not known previously in the strains, including genes for the biosynthesis of the siderophores achromobactin and pseudomonine and the antibiotic 2-hexyl-5-propyl-alkylresorcinol; novel bacteriocins; type II, III, and VI secretion systems; and insect toxins. Certain gene clusters, such as those for two type III secretion systems, are present only in specific sub-clades, suggesting vertical inheritance. Almost all of the genes associated with multitrophic interactions map to genomic regions present in only a subset of the strains or unique to a specific strain. To explore the evolutionary origin of these genes, we mapped their distributions relative to the locations of mobile genetic elements and repetitive extragenic palindromic (REP elements in each genome. The mobile genetic elements and many strain-specific genes fall into regions devoid of REP elements (i.e., REP deserts and regions displaying atypical tri-nucleotide composition, possibly indicating relatively recent acquisition of these loci. Collectively, the results of this study highlight the enormous heterogeneity of the P. fluorescens group and the importance of the variable genome in tailoring

  4. Nacre-mimic Reinforced Ag@reduced Graphene Oxide-Sodium Alginate Composite Film for Wound Healing.

    Science.gov (United States)

    Yan, Xu; Li, Fei; Hu, Kang-Di; Xue, Jingzhe; Pan, Xiao-Feng; He, Tao; Dong, Liang; Wang, Xiang-Ying; Wu, Ya-Dong; Song, Yong-Hong; Xu, Wei-Ping; Lu, Yang

    2017-10-23

    With the emerging of drug-resistant bacterial and fungal pathogens, there raise the interest of utilizing versatile antimicrobial biomaterials to treat the acute wound. Herein, we report the spraying mediated assembly of a bio-inspired Ag@reduced graphene-sodium alginate (AGSA) composite film for effective wound healing. The obtained film displayed lamellar microstructures similar to the typical "brick-and-mortar" structure in nacre. In this nacre-mimic structure, there are abundant interfacial interactions between nanosheets and polymeric matrix, leading to remarkable reinforcement. As a result, the tensile strength, toughness and Young's modulus have been improved 2.8, 2.3 and 2.7 times compared with pure sodium alginate film, respectively. In the wound healing study, the AGSA film showed effective antimicrobial activities towards Pseudomonas aeruginosa, Escherichia coli and Candida albicans, demonstrating the ability of protecting wound from pathogenic microbial infections. Furthermore, in vivo experiments on rats suggested the effect of AGSA film in promoting the recovery of wound sites. According to MTT assays, heamolysis evaluation and in vivo toxicity assessment, the composite film could be applied as a bio-compatible material in vitro and in vivo. Results from this work indicated such AGSA film has promising performance for wound healing and suggested great potential for nacre-mimic biomaterials in tissue engineering applications.

  5. Pseudomonas aeruginosa biofilm infections

    DEFF Research Database (Denmark)

    Tolker-Nielsen, Tim

    2014-01-01

    Bacteria in natural, industrial and clinical settings predominantly live in biofilms, i.e., sessile structured microbial communities encased in self-produced extracellular matrix material. One of the most important characteristics of microbial biofilms is that the resident bacteria display...... a remarkable increased tolerance toward antimicrobial attack. Biofilms formed by opportunistic pathogenic bacteria are involved in devastating persistent medical device-associated infections, and chronic infections in individuals who are immune-compromised or otherwise impaired in the host defense. Because...... the use of conventional antimicrobial compounds in many cases cannot eradicate biofilms, there is an urgent need to develop alternative measures to combat biofilm infections. The present review is focussed on the important opportunistic pathogen and biofilm model organism Pseudomonas aeruginosa. Initially...

  6. Magnesium Oxide Nanoparticles Reinforced Electrospun Alginate-Based Nanofibrous Scaffolds with Improved Physical Properties

    Directory of Open Access Journals (Sweden)

    R. T. De Silva

    2017-01-01

    Full Text Available Mechanically robust alginate-based nanofibrous scaffolds were successfully fabricated by electrospinning method to mimic the natural extracellular matrix structure which benefits development and regeneration of tissues. Alginate-based nanofibres were electrospun from an alginate/poly(vinyl alcohol (PVA polyelectrolyte complex. SEM images revealed the spinnability of the complex composite nanofibrous scaffolds, showing randomly oriented, ultrafine, and virtually defects-free alginate-based/MgO nanofibrous scaffolds. Here, it is shown that an alginate/PVA complex scaffold, blended with near-spherical MgO nanoparticles (⌀ 45 nm at a predetermined concentration (10% (w/w, is electrospinnable to produce a complex composite nanofibrous scaffold with enhanced mechanical stability. For the comparison purpose, chemically cross-linked electrospun alginate-based scaffolds were also fabricated. Tensile test to rupture revealed the significant differences in the tensile strength and elastic modulus among the alginate scaffolds, alginate/MgO scaffolds, and cross-linked alginate scaffolds (P<0.05. In contrast to cross-linked alginate scaffolds, alginate/MgO scaffolds yielded the highest tensile strength and elastic modulus while preserving the interfibre porosity of the scaffolds. According to the thermogravimetric analysis, MgO reinforced alginate nanofibrous scaffolds exhibited improved thermal stability. These novel alginate-based/MgO scaffolds are economical and versatile and may be further optimised for use as extracellular matrix substitutes for repair and regeneration of tissues.

  7. In vitro adhesion of human dermal fibroblasts on iron cross-linked alginate films

    International Nuclear Information System (INIS)

    Machida-Sano, Ikuko; Namiki, Hideo; Matsuda, Yasushi

    2009-01-01

    We evaluated the potential of alginate film incorporating ferric ions as a gelling agent (Fe-alginate) in comparison with that incorporating calcium ions (Ca-alginate) as a scaffold for culturing normal human dermal fibroblasts (NHDF). NHDF adhered to Fe-alginate and proliferated well, but no growth of the cells was observed on Ca-alginate. Since vitronectin and fibronectin play pivotal roles in cellular adhesion, their participation in NHDF behavior on alginate surfaces was investigated. We found that vitronectin was a critical element for initial attachment and spreading of NHDF on Fe-alginate. The surface properties of both alginate films were characterized in terms of protein adsorption ability and surface wettability, and it was revealed that Fe-alginate film adsorbed a significantly higher amount of proteins, including vitronectin and fibronectin, and had a higher surface hydrophobicity than Ca-alginate film. Moreover, under serum-free conditions, only a small number of NHDF were able to attach to the surface of Fe-alginate. Fe-alginate appeared to provide an appropriate surface for cellular attachment by adsorption of serum proteins such as vitronectin. These results suggest that Fe-alginate can serve as a scaffold for human fibroblasts and may be useful for tissue engineering research and other biomedical applications.

  8. Selective inhibition of Erwinia amylovora by the herbicidally active germination-arrest factor (GAF) produced by Pseudomonas bacteria.

    Science.gov (United States)

    Halgren, A; Azevedo, M; Mills, D; Armstrong, D; Thimmaiah, M; McPhail, K; Banowetz, G

    2011-10-01

     The germination-arrest factor (GAF) produced by Pseudomonas fluorescens WH6, and identified as 4-formylaminooxyvinylglycine, specifically inhibits the germination of a wide range of grassy weeds. This study was undertaken to determine whether GAF has antimicrobial activity in addition to its inhibitory effects on grass seed germination. Culture filtrate from Ps. fluorescens WH6 had little or no effect on 17 species of bacteria grown in Petri dish lawns, but the in vitro growth of Erwinia amylovora, the causal agent of the disease of orchard crops known as fire blight, was strongly inhibited by the filtrate. The anti-Erwinia activity of WH6 culture filtrate was shown to be due to its GAF content, and a commercially available oxyvinylglycine, 4-aminoethoxyvinylglycine (AVG), exhibited anti-Erwinia activity similar to that of GAF. The effects of GAF on Erwinia were reversed by particular amino acids. The biological properties of GAF include a rather specific antimicrobial activity against Erw. amylovora. This may be a general property of oxyvinylglycines as AVG exhibited similar activity. The ability of particular amino acids to reverse GAF inhibition is consistent with a potential effect of this compound on the activity of aminotransferases. The results presented here demonstrate a novel antimicrobial activity of oxyvinylglycines and suggest that GAF and/or GAF-producing bacteria may have potential for the control of fire blight. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology. No claim to US Government works.

  9. ANTAGONISTIC POTENTIAL OF FLUORESCENT Pseudomonas ...

    African Journals Online (AJOL)

    Prof. Adipala Ekwamu

    GROWTH OF TOMATO CHALLENGED WITH PHTOPATHOGENS ... This study focused on the antagonistic potential of fluorescent Pseudomonas in vitro, and its inoculation effect on growth .... the 5 days old culture in starch agar with Lugol's.

  10. Efecto de la aplicación de pseudomonas fluorescentes en las propiedades físico-químicas y biológicas de suelo : balance nutricional y estimulación de hormonas

    OpenAIRE

    Martínez Moreno, Joaquín Ignacio

    2015-01-01

    El objetivo de esta propuesta es el estudio del efecto de la aplicación de un fitofortificante (bacterias, Pseudomonas fluorescens) en la evolución tanto de las propiedades físico-químicas y microbiológicas/bioquímicas del suelo como en la estimulación de hormonas del crecimiento vegetal, así como estudiar su efecto en el balance nutricional, la calidad y la producción de los cultivos de maíz y melón. Escuela Técnica superior de Ingeniería Agronómica Universidad Politécnica ...

  11. Characterization of Extracellular Polymeric Substances Produced by Pseudomonas fragi Under Air and Modified Atmosphere Packaging.

    Science.gov (United States)

    Wang, Guang-Yu; Ma, Fang; Wang, Hu-Hu; Xu, Xing-Lian; Zhou, Guang-Hong

    2017-09-01

    Extracellular polymeric substances (EPS) play an important role in bacterial biochemical properties. The characteristics of EPS from 2 strains of Pseudomonas fragi cultured in meat aerobically (control) and in modified atmosphere packaging (MAP) were studied. The amount and components of EPS, the surface properties, and the effect on biofilm formation of several spoilage organisms were evaluated. The results showed that MAP inhibited the growth of the P. fragi strains. Compared with the control, more loose and less bound EPS (containing protein and carbohydrate) were produced by P. fragi in MAP samples. MAP also caused increased cell autoaggregation and surface hydrophobicity. After the removal of the EPS, the surface property changes were strain-dependent, suggesting that membrane compositions were also changed. In addition, the EPS displayed significant antibiofilm activity on Pseudomonas fluorescens and Serratia liquefaciens. In conclusion, P. fragi strains not only modified the amount, components, and surface properties of EPS but also changed the cell membrane compositions to adapt to MAP stress. Moreover, EPS may play an important role in microbial community competitions. © 2017 Institute of Food Technologists®.

  12. Cytotoxicity of alginate for orthodontic use

    Directory of Open Access Journals (Sweden)

    Matheus Melo Pithon

    2012-12-01

    Full Text Available OBJECTIVE: To evaluate the cytotoxicity of three different alginate impression materials for orthodontic use. METHODS: Three different brands of alginate were divided into three groups, namely, Group JCO (Jeltrate Chromatic Ortho, OP (Orthoprint and CO (Cavex Orthotrace. Three control groups were also included: Group C+ (positive control, consisting of detergent Tween 80; Group C- (negative control, consisting of PBS, and Group CC (cell control, consisting of cells not exposed to any material. After manipulating the materials according to the respective manufacturer instructions, samples were made with the use of silicon rings. Then the samples were immersed in Eagle's minimum essential medium (MEM for 2 minutes. The supernatants were then removed and brought into direct contact with L929 fibroblasts. After exposure to the medium, the cells were incubated for 24 hours. Then 100 µl of 0.01% neutral red dye were added. The cells were incubated again for 3 hours so that the dye could be absorbed. After this 3-hour period, the cells were fixed to perform the viable cell count, using a spectrophotometer (BioTek, Winooski, Vermont, USA at a wavelength of 492 nm. RESULTS: Statistical differences were found when Groups CC and C- were compared with the other experimental groups. Group JCO had the highest cytotoxicity, followed by Groups OP and CO. CONCLUSION: Based on the results obtained in this work, it was concluded that all alginate impression materials are potentially cytotoxic.OBJETIVO: avaliar a citotoxicidade de três diferentes alginatos de uso ortodôntico. MÉTODOS: foram avaliados três diferentes alginatos divididos em três grupos, denominados grupo JCO (Jeltrate Chromatic Ortho, OP (Orthoprint e CO (Carrex Orthotrace. Três grupos controle também participaram: controle + (C+, constituído pelo detergente celular Tween 80; controle - (C- PBS; e controle de célula (CC onde as células não foram expostas a nenhum material. Após manipula

  13. Properties of alginate fiber spun-dyed with fluorescent pigment dispersion.

    Science.gov (United States)

    Wang, Ping; Tawiah, Benjamin; Tian, Anli; Wang, Chunxia; Zhang, Liping; Fu, Shaohai

    2015-03-15

    Spun-dyed alginate fiber was prepared by the spun-dyeing method with the mixture of fluorescent pigment dispersion and sodium alginate fiber spinning solution, and its properties were characterized by SEM, TGA, DSC, and XRD. The results indicate that fluorescent pigment dispersion prepared with esterified poly (styrene-alt maleic acid) had excellent compatibility with sodium alginate fiber spinning solution, and small amount of fluorescent pigment could reduce the viscosity of spun-dyed spinning solutions. SEM photo of spun-dyed alginate fiber indicated that fewer pigment particles deposited on its surface. TGA, DSC, and XRD results suggested that thermal properties and crystal phase of spun-dyed alginate fibers had slight changes compared to the original alginate fibers. The fluorescence intensity of spun-dyed alginate fiber reached its maximum when the content of fluorescent pigment was 4%. The spun-dyed alginate fiber showed excellent rubbing and washing fastness. Copyright © 2014 Elsevier Ltd. All rights reserved.

  14. The Utilization of Additional Cassava Starch (Manihot Utilisima) for Alginate Dental Impression Material

    OpenAIRE

    Ali Noerdin; Bambang Irawan; Mirna Febriani

    2003-01-01

    In Indonesia alginate which is a common impression material used in dentistry is still imported. Since the economic crisis in 1998 the alginate price becoming four times more expensive. This situation resulted in efforts to modify the commercial alginate as had been conducted by a dentist in South Sumatera province in Indonesia. He who had added cassava starch into the commercial alginate used to make partial denture impression. The aim of this research is to investigate the effect of additio...

  15. Injectable hydrogels derived from phosphorylated alginic acid calcium complexes

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Han-Sem; Song, Minsoo, E-mail: minsoosong00@gmail.com; Lee, Eun-Jung; Shin, Ueon Sang, E-mail: usshin12@dankook.ac.kr

    2015-06-01

    Phosphorylation of sodium alginate salt (NaAlg) was carried out using H{sub 3}PO{sub 4}/P{sub 2}O{sub 5}/Et{sub 3}PO{sub 4} followed by acid–base reaction with Ca(OAc){sub 2} to give phosphorylated alginic acid calcium complexes (CaPAlg), as a water dispersible alginic acid derivative. The modified alginate derivatives including phosphorylated alginic acid (PAlg) and CaPAlg were characterized by nuclear magnetic resonance spectroscopy for {sup 1}H, and {sup 31}P nuclei, high resolution inductively coupled plasma optical emission spectroscopy, Fourier transform infrared spectroscopy, and thermogravimetric analysis. CaPAlg hydrogels were prepared simply by mixing CaPAlg solution (2 w/v%) with NaAlg solution (2 w/v%) in various ratios (2:8, 4:6, 6:4, 8:2) of volume. No additional calcium salts such as CaSO{sub 4} or CaCl{sub 2} were added externally. The gelation was completed within about 3–40 min indicating a high potential of hydrogel delivery by injection in vivo. Their mechanical properties were tested to be ≤ 6.7 kPa for compressive strength at break and about 8.4 kPa/mm for elastic modulus. SEM analysis of the CaPAlg hydrogels showed highly porous morphology with interconnected pores of width in the range of 100–800 μm. Cell culture results showed that the injectable hydrogels exhibited comparable properties to the pure alginate hydrogel in terms of cytotoxicity and 3D encapsulation of cells for a short time period. The developed injectable hydrogels showed suitable physicochemical and mechanical properties for injection in vivo, and could therefore be beneficial for the field of soft tissue engineering. - Highlights: • Preparation of water-soluble alginic acid complexes with calcium phosphate • Self-assembly of the phosphorylated alginic acid calcium complexes with sodium alginate • Preparation of injectable hydrogels with diverse gelation times within about 3–40 min.

  16. Novel experimental Pseudomonas aeruginosa lung infection model mimicking long-term host-pathogen interactions in cystic fibrosis

    DEFF Research Database (Denmark)

    Moser, Claus; van Gennip, Maria; Bjarnsholt, Thomas

    2009-01-01

    Moser C, van Gennip M, Bjarnsholt T, Jensen PO, Lee B, Hougen HP, Calum H, Ciofu O, Givskov M, Molin S, Hoiby N. Novel experimental Pseudomonas aeruginosa lung infection model mimicking long-term host-pathogen interactions in cystic fibrosis. APMIS 2009; 117: 95-107. The dominant cause of premature...... death in patients suffering from cystic fibrosis (CF) is chronic lung infection with Pseudomonas aeruginosa. The chronic lung infection often lasts for decades with just one clone. However, as a result of inflammation, antibiotic treatment and different niches in the lungs, the clone undergoes...... and 2003) of the chronic lung infection of one CF patient using the seaweed alginate embedment model. The results showed that the non-mucoid clones reduced their virulence over time, resulting in faster clearing of the bacteria from the lungs, improved pathology and reduced pulmonary production...

  17. A VERSATILE ALGINATE DROPLET GENERATOR APPLICABLE FOR MICROENCAPSULATION OF PANCREATIC-ISLETS

    NARCIS (Netherlands)

    WOLTERS, GHJ; FRITSCHY, WM; GERRITS, D; VANSCHILFAGAARDE, R

    1992-01-01

    Alginate beads for immunoisolation of pancreatic islets by microencapsulation should be small, smooth, and spherical in order to ensure that around the islets a strong alginate-polylysine-alginate capsule will be formed with optimal biocompatibility and diffusion of nutrients and hormones. However,

  18. Improved biocompatibility but limited graft survival after purification of alginate for microencapsulation of pancreatic islets

    NARCIS (Netherlands)

    DeVos, P; DeHaan, BJ; Wolters, GHJ; Strubbe, JH; VanSchilfgaarde, R; van Schilfgaarde, P.

    Graft failure of alginate-polylysine microencapsulated islets is often interpreted as the consequence of a non-specific foreign body reaction against the microcapsules, initiated by impurities present in crude alginate. The aim of the present study was to investigate if purification of the alginate

  19. Three-dimensional alginate spheroid culture system of murine osteosarcoma.

    Science.gov (United States)

    Akeda, Koji; Nishimura, Akinobu; Satonaka, Haruhiko; Shintani, Ken; Kusuzaki, Katsuyuki; Matsumine, Akihiko; Kasai, Yuichi; Masuda, Koichi; Uchida, Atsumasa

    2009-11-01

    Osteosarcoma (OS) is the most common primary malignant tumor of the bone and often forms pulmonary metastases, which are the most important prognostic factor. For further elucidation of the mechanism underlying the progression and metastasis of human OS, a culture system mimicking the microenvironment of the tumor in vivo is needed. We report a novel three-dimensional (3D) alginate spheroid culture system of murine osteosarcoma. Two different metastatic clones, the parental Dunn and its derivative line LM8, which has a higher metastatic potential to the lungs, were encapsulated in alginate beads to develop the 3D culture system. The beads containing murine OS cells were also transplanted into mice to determine their metastatic potential in vivo. In this culture system, murine OS cells encapsulated in alginate beads were able to grow in a 3D structure with cells detaching from the alginate environment. The number of detaching cells was higher in the LM8 cell line than the Dunn cell line. In the in vivo alginate bead transplantation model, the rate of pulmonary metastasis was higher with LM8 cells compared with that of Dunn cells. The cell characteristics and kinetics in this culture system closely reflect the original malignant potential of the cells in vivo.

  20. Applications of Alginate-Based Bioinks in 3D Bioprinting

    Directory of Open Access Journals (Sweden)

    Eneko Axpe

    2016-11-01

    Full Text Available Three-dimensional (3D bioprinting is on the cusp of permitting the direct fabrication of artificial living tissue. Multicellular building blocks (bioinks are dispensed layer by layer and scaled for the target construct. However, only a few materials are able to fulfill the considerable requirements for suitable bioink formulation, a critical component of efficient 3D bioprinting. Alginate, a naturally occurring polysaccharide, is clearly the most commonly employed material in current bioinks. Here, we discuss the benefits and disadvantages of the use of alginate in 3D bioprinting by summarizing the most recent studies that used alginate for printing vascular tissue, bone and cartilage. In addition, other breakthroughs in the use of alginate in bioprinting are discussed, including strategies to improve its structural and degradation characteristics. In this review, we organize the available literature in order to inspire and accelerate novel alginate-based bioink formulations with enhanced properties for future applications in basic research, drug screening and regenerative medicine.

  1. Immobilization of myoglobin in sodium alginate composite membranes

    Directory of Open Access Journals (Sweden)

    Katia Cecília de Souza Figueiredo

    2015-06-01

    Full Text Available AbstractThe immobilization of myoglobin in sodium alginate films was investigated with the aim of evaluating the protein stability in an ionic polymeric matrix. Myoglobin was chosen due to the resemblance to each hemoglobin tetramer. Sodium alginate, being a natural polysaccharide, was selected as the polymeric matrix because of its chemical structure and film-forming ability. To improve the mechanical resistance of sodium alginate films, the polymer was deposited over the surface of a cellulose acetate support by means of ultrafiltration. The ionic crosslink of sodium alginate was investigated by calcium ions. Composite membrane characterization comprised water swelling tests, water flux, SEM images and UV-visible spectroscopy. The electrostatic interaction between the protein and the polysaccharide did not damage the UV-visible pattern of native myoglobin. A good affinity between sodium alginate and cellulose acetate was observed. The top layer of the dense composite membrane successfully immobilized Myoglobin, retaining the native UV-visible pattern for two months.

  2. Use of antacids, alginates and proton pump inhibitors

    DEFF Research Database (Denmark)

    Lødrup, Anders; Reimer, Christine; Bytzer, Peter

    2014-01-01

    : A cross-sectional survey was conducted in an internet panel representative of the Danish adult population in 2012. Data queried included antacid/alginate and PPI use, reason for therapy, co-medication, and presence of upper gastrointestinal symptoms. Long-term PPI use was defined as using PPI ≥1/3...... of the last year (∼120 days). Risk of long-term PPI use was estimated by logistic regression. RESULTS: A total of 18,223 people received the questionnaire, of which 52% (9390) responded. Antacid/alginate use was reported by 23%; 16% reported use of only antacid/alginate. PPI use was reported by 13.6%; 6....../e-mail, using co-medication, and having started on PPI for several reasons. Combination of antacid/alginate and PPI was reported by approximately 50% of those on therapy with weekly or daily symptoms. CONCLUSION: 23% of Danish adults were using antacids or alginates and 14% were using PPI, of which one...

  3. Bacteriocinogenicity and production of pyocins from Pseudomonas ...

    African Journals Online (AJOL)

    ), P. fluorescens (13) and P. stutzeri (3) were isolated from Nigerian patients with burns, wound and skin infections, UTI, diarrhoea and eye infections. 81.5, 55.6, 76.9 and 33.3% of these species produced pyocins at a range of 410 – 670 g/ml.

  4. Nanostructured magnetic alginate composites for biomedical applications

    Energy Technology Data Exchange (ETDEWEB)

    Bedê, Pedro Marins; Silva, Marcelo Henrique Prado da; Figueiredo, André Ben-Hur da Silva, E-mail: marceloprado@ime.eb.br [Instituto Militar de Engenharia (IME), Rio de Janeiro, RJ (Brazil); Finotelli, Priscilla Vanessa [Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, RJ (Brazil). Faculdade de Farmácia

    2017-07-01

    This is a study of the preparation and characterization of polymeric-magnetic nanoparticles. The nanoparticles used were magnetite (Fe{sub 3}O{sub 4} ) and the chosen polymers were alginate and chitosan. Two types of samples were prepared: uncoated magnetic nanoparticles and magnetic nanoparticles encapsulated in polymeric matrix. The samples were analyzed by XRD, light scattering techniques, TEM, and magnetic SQUID. The XRD patterns identified magnetite (Fe{sub 3}O{sub 4} ) as the only crystalline phase. TEM analyses showed particle sizes between 10 and 20nm for magnetite, and 15 and 30nm for the encapsulated magnetite. The values of magnetization ranged from 75 to 100emu/g for magnetite nanoparticles, and 8 to 12emu/g for coated with chitosan, at different temperatures of 20K and 300K. The saturation of both samples was in the range of 49 to 50KOe. Variations of results between the two kinds of samples were attributed to the encapsulation of magnetic nanoparticles by the polymers. (author)

  5. Two small RNAs, CrcY and CrcZ, act in concert to sequester the Crc global regulator in Pseudomonas putida, modulating catabolite repression.

    Science.gov (United States)

    Moreno, Renata; Fonseca, Pilar; Rojo, Fernando

    2012-01-01

    The Crc protein is a translational repressor that recognizes a specific target at some mRNAs, controlling catabolite repression and co-ordinating carbon metabolism in pseudomonads. In Pseudomonas aeruginosa, the levels of free Crc protein are controlled by CrcZ, a sRNA that sequesters Crc, acting as an antagonist. We show that, in Pseudomonas putida, the levels of free Crc are controlled by CrcZ and by a novel 368 nt sRNA named CrcY. CrcZ and CrcY, which contain six potential targets for Crc, were able to bind Crc specifically in vitro. The levels of CrcZ and CrcY were low under conditions generating a strong catabolite repression, and increased strongly when catabolite repression was absent. Deletion of either crcZ or crcY had no effect on catabolite repression, but the simultaneous absence of both sRNAs led to constitutive catabolite repression that compromised growth on some carbon sources. Overproduction of CrcZ or CrcY significantly reduced repression. We propose that CrcZ and CrcY act in concert, sequestering and modulating the levels of free Crc according to metabolic conditions. The CbrA/CbrB two-component system activated crcZ transcription, but had little effect on crcY. CrcY was detected in P. putida, Pseudomonas fluorescens and Pseudomonas syringae, but not in P. aeruginosa. © 2011 Blackwell Publishing Ltd.

  6. The Drug Release Profile from Calcium-induced Alginate Gel Beads Coated with an Alginate Hydrolysate

    Directory of Open Access Journals (Sweden)

    Susumu Kawashima

    2007-11-01

    Full Text Available Calcium-induced alginate gel bead (Alg-Ca coated with an alginate hydrolysate(Alg, e.g. the guluronic acid block (GB was prepared and the model drug, hydrocortisonerelease profiles were investigated under simulated gastrointestinal conditions. Theirmolecular weights were one sixth or one tenth that of Alg and the diffraction patterns of thehydrolysates resembled that of Alg. The drug release rate from Alg-Ca coated with GBapparently lowered than that of Alg-Ca (coating-free in the gastric juice (pH1.2. And thecoating did not resist the disintegration of Alg-Ca in the intestinal juice (pH 6.8 and thegel erosion accelerated the drug release. On the other hand, for the coated Alg-Cacontaining chitosan, the drug release showed zero-order kinetics without rapid erosion ofAlg-Ca. The drug release rate from Alg-Ca was able to be controlled by the coating andmodifying the composition of the gel matrix.

  7. Phosphorylcholine Phosphatase: A Peculiar Enzyme of Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Carlos Eduardo Domenech

    2011-01-01

    Full Text Available Pseudomonas aeruginosa synthesizes phosphorylcholine phosphatase (PchP when grown on choline, betaine, dimethylglycine or carnitine. In the presence of Mg2+ or Zn2+, PchP catalyzes the hydrolysis of p-nitrophenylphosphate (p-NPP or phosphorylcholine (Pcho. The regulation of pchP gene expression is under the control of GbdR and NtrC; dimethylglycine is likely the metabolite directly involved in the induction of PchP. Therefore, the regulation of choline metabolism and consequently PchP synthesis may reflect an adaptive response of P. aeruginosa to environmental conditions. Bioinformatic and biochemistry studies shown that PchP contains two sites for alkylammonium compounds (AACs: one in the catalytic site near the metal ion-phosphoester pocket, and another in an inhibitory site responsible for the binding of the alkylammonium moiety. Both sites could be close to each other and interact through the residues 42E, 43E and 82YYY84. Zn2+ is better activator than Mg2+ at pH 5.0 and it is more effective at alleviating the inhibition produced by the entry of Pcho or different AACs in the inhibitory site. We postulate that Zn2+ induces at pH 5.0 a conformational change in the active center that is communicated to the inhibitory site, producing a compact or closed structure. However, at pH 7.4, this effect is not observed because to the hydrolysis of the [Zn2+L2−1L20(H2O2] complex, which causes a change from octahedral to tetrahedral in the metal coordination geometry. This enzyme is also present in P. fluorescens, P. putida, P. syringae, and other organisms. We have recently crystallized PchP and solved its structure.

  8. Application of neutral electrolyzed water to disinfection of alginate impression.

    Science.gov (United States)

    Nagamatsu, Yuki; Chen, Ker-Kong; Nagamatsu, Hiroshi; Kozono, Yoshio; Shimizu, Hiroshi

    2016-01-01

    Neutral electrolyzed water was developed with new concepts of long-term good durability and minimum corrosiveness to metal in addition to its excellent bactericidal activities similar to acid type of electrolyzed waters. The present study examined the bactericidal effects of the neutral electrolyzed water on disinfection of the alginate impression of a dental arch model contaminated by bacteria. Only 1-min immersion in neutral electrolyzed water could sufficiently disinfect the alginate impression including the metallic tray under ultrasonic with no significant differences from acid electrolyzed waters. No bactericidal effects were found in any electrolyzed water when used as mixing water. Considering the advantages and disadvantages of each electrolyzed water in a comprehensive way, it was suggested that neutral electrolyzed water may be the most appropriate for the disinfection of alginate impression.

  9. Cloning of genes required for hypersensitivity and pathogenicity in Pseudomonas syringae pv. aptata.

    Science.gov (United States)

    Minardi, P

    1995-01-01

    A genomic library of Pseudomonas syringae pv. aptata strain NCPPB 2664, which causes bacterial blight of sugar beet, lettuce and other plants, was constructed in the cosmid vector pCPP31. The 13.4 kb EcoRI fragment of the cosmid pHIR11, containing the hrp (hypersensitive response and pathogenicity) gene cluster of the closely related bacterium Pseudomonas syringae pv. syringae strain 61, was used as a probe to identify a homologous hrp gene cluster in P. syringae pv. aptata. Thirty of 2500 cosmid clones, screened by colony hybridization, gave a strong hybridization signal with the probe, but none of these conferred to the non-pathogenic bacterium, Pseudomonas fluorescens, the ability to elicit the hypersensitive response (HR) in tobacco. Southern blot analysis of EcoRI-digested genomic DNA of P. syringae pv. aptata showed hybridizing bands of 12 kb and 4.4 kb. Only a 12 kb fragment hybridized in digests of the cosmids. Cosmid clone pCPP1069 was mutagenized with Tn10-minitet and marker-exchanged into the genome of P. syringae pv. aptata. Three resulting prototrophic mutant strains failed to elicit the HR in tobacco and to cause disease in lettuce. The DNA flanking the Tn10-minitet insertions from mutated derivatives of pCPP1069 hybridized with the 10.6 kb Bg/II fragment of pHIR11. These results indicate that P. syringae pv. aptata harbours hrp genes that are similar to, but arranged differently from, homologous hrp genes of P. syringae pv. syringae.

  10. Bioactive apatite incorporated alginate microspheres with sustained drug-delivery for bone regeneration application

    Energy Technology Data Exchange (ETDEWEB)

    Li, Haibin; Jiang, Fei; Ye, Song; Wu, Yingying; Zhu, Kaiping; Wang, Deping, E-mail: wdpshk@tongji.edu.cn

    2016-05-01

    The strontium-substituted hydroxyapatite microspheres (SrHA) incorporated alginate composite microspheres (SrHA/Alginate) were prepared via adding SrHA/alginate suspension dropwise into calcium chloride solution, in which the gel beads were formed by means of crosslinking reaction. The structure, morphology and in vitro bioactivity of the composite microspheres were studied by using XRD, SEM and EDS methods. The biological behaviors were characterized and analyzed through inductively coupled plasma optical emission spectroscopy (ICP-OES), CCK-8, confocal laser microscope and ALP activity evaluations. The experimental results indicated that the synthetic SrHA/Alginate showed similar morphology to the well-known alginate microspheres (Alginate) and both of them possessed a great in vitro bioactivity. Compared with the control Alginate, the SrHA/Alginate enhanced MC3T3-E1 cell proliferation and ALP activity by releasing osteoinductive and osteogenic Sr ions. Furthermore, vancomycin was used as a model drug to investigate the drug release behaviors of the SrHA/Alginate, Alginate and SrHA. The results suggested that the SrHA/Alginate had a highest drug-loading efficiency and best controlled drug release properties. Additionally, the SrHA/Alginate was demonstrated to be pH-sensitive as well. The increase of the pH value in phosphate buffer solution (PBS) accelerated the vancomycin release. Accordingly, the multifunctional SrHA/Alginate can be applied in the field of bioactive drug carriers and bone filling materials. - Highlights: • The pH-sensitive composite alginate beads incorporating Sr-doped HA microspheres (SrHA) have been prepared. • The incorporation of the SrHA enhanced the drug loading and release properties of the alginate microspheres. • The composite microspheres showed excellent osteogenic effect by releasing osteogenic Sr ions.

  11. Production, deformation and mechanical investigation of magnetic alginate capsules

    Science.gov (United States)

    Zwar, Elena; Kemna, Andre; Richter, Lena; Degen, Patrick; Rehage, Heinz

    2018-02-01

    In this article we investigated the deformation of alginate capsules in magnetic fields. The sensitivity to magnetic forces was realised by encapsulating an oil in water emulsion, where the oil droplets contained dispersed magnetic nanoparticles. We solved calcium ions in the aqueous emulsion phase, which act as crosslinking compounds for forming thin layers of alginate membranes. This encapsulating technique allows the production of flexible capsules with an emulsion as the capsule core. It is important to mention that the magnetic nanoparticles were stable and dispersed throughout the complete process, which is an important difference to most magnetic alginate-based materials. In a series of experiments, we used spinning drop techniques, capsule squeezing experiments and interfacial shear rheology in order to determine the surface Young moduli, the surface Poisson ratios and the surface shear moduli of the magnetically sensitive alginate capsules. In additional experiments, we analysed the capsule deformation in magnetic fields. In spinning drop and capsule squeezing experiments, water droplets were pressed out of the capsules at elevated values of the mechanical load. This phenomenon might be used for the mechanically triggered release of water-soluble ingredients. After drying the emulsion-filled capsules, we produced capsules, which only contained a homogeneous oil phase with stable suspended magnetic nanoparticles (organic ferrofluid). In the dried state, the thin alginate membranes of these particles were rather rigid. These dehydrated capsules could be stored at ambient conditions for several months without changing their properties. After exposure to water, the alginate membranes rehydrated and became flexible and deformable again. During this swelling process, water diffused back in the capsule. This long-term stability and rehydration offers a great spectrum of different applications as sensors, soft actuators, artificial muscles or drug delivery systems.

  12. The Use of Alginate in Lemon Extract Effervescent Powder Production

    Directory of Open Access Journals (Sweden)

    Murdinah

    2015-11-01

    Full Text Available Study on the use of alginate in lemon (Citrus medica var lemon extract effervescent powder production has conducted. The aims of the research are to determine the optimum concentration of alginate used in lemon extract effervescent powder to produced best product and acceptance consumen.The lemon extract effervescent powder formula consisted of lemon extract powder, sucrose, aspartame, salt and effervescent mix (citric acid-tartrat acid-sodium bicarbonat. The alginate used in this study was extracted from Sargassum filipendula sea weed. The concentration of alginate used in lemon effervescent powder production was varied from 1; 2; 3 and 4%. The parameters observed to see the quality of the product were moisture content, ash content, pH, viscosity and organoleptic value (flavor, taste, viscosity, effec effervescent, effect sparkle and acceptance. Analysis of dietary fiber, sugar content, vitamin C content, total titratable acids, TPC and E.Coli to the best product. The result showed that the higher the concentration of alginate used in lemon effervescent powder production, the higher viscousness and the lower the organoleptic value. The optimum concentration of alginate used in the lemon extract effervescent powder processing was 1%. The characteristic this product 7.60% moisture content, 0.86% insoluble dietary fiber , 7.92% soluble dietary fiber, 3.74% sugar content, 55,26 mg/100 g vitamin C, 134.15 mL 0.1 NaOH/100 mL total titratable acids, 20 cPs viscosity, <2.5x102 coloni/mL TPC and E.Coli negative.

  13. Alginate Biosynthesis in Azotobacter vinelandii: Overview of Molecular Mechanisms in Connection with the Oxygen Availability

    Directory of Open Access Journals (Sweden)

    Ivette Pacheco-Leyva

    2016-01-01

    Full Text Available The Gram-negative bacterium Azotobacter vinelandii can synthetize the biopolymer alginate that has material properties appropriate for plenty of applications in industry as well as in medicine. In order to settle the foundation for improving alginate production without compromising its quality, a better understanding of the polymer biosynthesis and the mechanism of regulation during fermentation processes is necessary. This knowledge is crucial for the development of novel production strategies. Here, we highlight the key aspects of alginate biosynthesis that can lead to producing an alginate with specific material properties with particular focus on the role of oxygen availability linked with the molecular mechanisms involved in the alginate production.

  14. Development of a Novel Alginate-Based Pleural Sealant

    Science.gov (United States)

    2016-07-01

    thoracotomy is performed on a anesthetized ventilated live rat (a), an incision is made to induce pleural air leak (b), and sealant material [ liquid ...AWARD NUMBER: W81XWH-15-1-0107 TITLE: Development of a Novel Alginate-Based Pleural Sealant PRINCIPAL INVESTIGATOR: Daniel J. Weiss MD PhD...SUBTITLE Development of a Novel Alginate-Based Pleural Sealant 5a. CONTRACT NUMBER 5b. GRANT NUMBER W81XWH-15-1-0107 5c. PROGRAM ELEMENT NUMBER 6. AUTHOR

  15. Effect of alginate in patients with GERD hiatal hernia matters.

    Science.gov (United States)

    Vardar, R; Keskin, M; Valitova, E; Bayrakci, B; Yildirim, E; Bor, S

    2017-10-01

    Alginate-based formulations are frequently used as add-on proton pump inhibitor (PPI) therapy to help control of heartburn and regurgitation. There are limited data regarding the mechanisms and effects of alginate-based formulations. We aimed to evaluate the effects of the sodium alginate intake and its likely temporal relations on intraesophageal reflux events by MII-pH in patients with and without hiatal hernia (HH). Fifty GERD patients (18 with HH, 32 without HH) with heartburn or regurgitation once a week or more common were included. After combined multichannel intraluminal impedance and pH-metry (MII-pH) had been performed, all patients were asked to eat the same standard meal (double cheeseburger, 1 banana, 100 g regular yoghurt, and 200 mL water with total energy value of 744 kcal: 37.6% of carbohydrates, 21.2% of proteins, and 41.2% of lipids) during two consecutive days. On separate random two consecutive days, all patients took 10 mL of sodium alginate (GA; Gaviscon Advance; Reckitt Benckiser Healthcare, Hull, UK) or 10 mL of water, 30 minutes after the refluxogenic meal. After eating refluxogenic meal, patients were examined ½ hour for basal conditions, 1 hour in upright, and 1 hour in supine positions. Alginate significantly decreased acid reflux after intake at the first hour in comparison to water in patients with HH (6.1 vs. 13.7, P = 0.004) and without HH (3.5 vs. 5.5, P = 0.001). Weakly acid reflux were increased at the first hour in patients with HH (3.4 vs. 1.3, P = 0.019) and without HH (1.7 vs. 5, P = 0.02) compared to water. There was no distinctive effect of alginate on the height of proximal migration of reflux events in patients with HH and without HH. Alginate decreases acid reflux events within a limited time period, especially at the first hour both in patients with and without HH. Alginate has no effect on the height of reflux events along the esophagus both in patients with and without HH. © The Authors 2017. Published by Oxford

  16. "Hot Tub Rash" and "Swimmer's Ear" (Pseudomonas)

    Science.gov (United States)

    Facts About “Hot Tub Rash” and “Swimmer’s Ear” (Pseudomonas) What is Pseudomonas and how can it affect me? Pseudomonas (sue-doh- ... a major cause of infections commonly known as “hot tub rash” and “swimmer’s ear.” This germ is ...

  17. Controlled release of metronidazole from composite poly-ε-caprolactone/alginate (PCL/alginate) rings for dental implants.

    Science.gov (United States)

    Lan, Shih-Feng; Kehinde, Timilehin; Zhang, Xiangming; Khajotia, Sharukh; Schmidtke, David W; Starly, Binil

    2013-06-01

    Dental implants provide support for dental crowns and bridges by serving as abutments for the replacement of missing teeth. To prevent bacterial accumulation and growth at the site of implantation, solutions such as systemic antibiotics and localized delivery of bactericidal agents are often employed. The objective of this study was to demonstrate a novel method of controlled localized delivery of antibacterial agents to an implant site using a biodegradable custom fabricated ring. The study involved incorporating a model antibacterial agent (metronidazole) into custom designed poly-ε-caprolactone/alginate (PCL/alginate) composite rings to produce the intended controlled release profile. The rings can be designed to fit around the body of any root form dental implants of various diameters, shapes and sizes. In vitro release studies indicate that pure (100%) alginate rings exhibited an expected burst release of metronidazole in the first few hours, whereas Alginate/PCL composite rings produced a medium burst release followed by a sustained release for a period greater than 4 weeks. By varying the PCL/alginate weight ratios, we have shown that we can control the amount of antibacterial agents released to provide the minimal inhibitory concentration (MIC) needed for adequate protection. The fabricated composite rings have achieved a 50% antibacterial agent release profile over the first 48 h and the remaining amount slowly released over the remainder of the study period. The PCL/alginate agent release characteristic fits the Ritger-Peppas model indicating a diffusion-based mechanism during the 30-day study period. The developed system demonstrates a controllable drug release profile and the potential for the ring to inhibit bacterial biofilm growth for the prevention of diseases such as peri-implantitis resulting from bacterial infection at the implant site. Copyright © 2013 Academy of Dental Materials. Published by Elsevier Ltd. All rights reserved.

  18. Evaluation of digital dental models obtained from dental cone-beam computed tomography scan of alginate impressions

    OpenAIRE

    Jiang, Tingting; Lee, Sang-Mi; Hou, Yanan; Chang, Xin; Hwang, Hyeon-Shik

    2016-01-01

    Objective To investigate the dimensional accuracy of digital dental models obtained from the dental cone-beam computed tomography (CBCT) scan of alginate impressions according to the time elapse when the impressions are stored under ambient conditions. Methods Alginate impressions were obtained from 20 adults using 3 different alginate materials, 2 traditional alginate materials (Alginoplast and Cavex Impressional) and 1 extended-pour alginate material (Cavex ColorChange). The impressions wer...

  19. Molecular cloning, purification, and characterization of a novel polyMG-specific alginate lyase responsible for alginate MG block degradation in Stenotrophomas maltophilia KJ-2

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Su In; Kim, Hee Sook [Kyungsung Univ., Busan (Korea, Republic of). Dept. of Food Science and Biotechnology; Choi, Sung Hee; Lee, Eun Yeol [Kyung Hee Univ., Gyeonggi-do (Korea, Republic of). Dept. of Chemical Engineering

    2012-09-15

    A gene for a polyMG-specific alginate lyase possessing a novel structure was identified and cloned from Stenotrophomas maltophilia KJ-2 by using PCR with homologous nucleotide sequences-based primers. The recombinant alginate lyase consisting of 475 amino acids was purified on Ni-Sepharose column and exhibited the highest activity at pH 8 and 40 C. Interestingly, the recombinant alginate lyase was expected to have a similar catalytic active site of chondroitin B lyase but did not show chondroitin lyase activity. In the test of substrate specificity, the recombinant alginate lyase preferentially degraded the glycosidic bond of polyMG-block than polyM-block and polyG-block. The chemical structures of the degraded alginate oligosaccharides were elucidated to have mannuronate (M) at the reducing end on the basis of NMR analysis, supporting that KJ-2 polyMG-specific alginate lyase preferably degraded the glycosidic bond in M-G linkage than that in G-M linkage. The KJ-2 polyMG-specific alginate lyase can be used in combination with other alginate lyases for a synergistic saccharification of alginate. (orig.)

  20. Initial characterization of a bolA homologue from Pseudomonas fluorescens indicates different roles for BolA-like proteins in P. fluorescens and Escherichia coli

    DEFF Research Database (Denmark)

    Koch, Birgit; Nybroe, Ole

    2006-01-01

    A expression. The mutant grew slower than the wild-type strain in minimal medium with L-serine as the sole nitrogen source, while growth rates were similar on a mixture of L-serine and L-cysteine. Reverse transcriptase polymerase chain reaction analysis indicated that the bolA homologue is the second gene...

  1. In vitro evaluation of alginate/halloysite nanotube composite scaffolds for tissue engineering

    International Nuclear Information System (INIS)

    Liu, Mingxian; Dai, Libing; Shi, Huizhe; Xiong, Sheng; Zhou, Changren

    2015-01-01

    In this study, a series of alginate/halloysite nanotube (HNTs) composite scaffolds were prepared by solution-mixing and freeze-drying method. HNTs are incorporated into alginate to improve both the mechanical and cell-attachment properties of the scaffolds. The interfacial interactions between alginate and HNTs were confirmed by the atomic force microscope (AFM), transmission electron microscope (TEM) and FTIR spectroscopy. The mechanical, morphological, and physico-chemical properties of the composite scaffolds were investigated. The composite scaffolds exhibit significant enhancement in compressive strength and compressive modulus compared with pure alginate scaffold both in dry and wet states. A well-interconnected porous structure with size in the range of 100–200 μm and over 96% porosity is found in the composite scaffolds. X-ray diffraction (XRD) result shows that HNTs are uniformly dispersed and partly oriented in the composite scaffolds. The incorporation of HNTs leads to increase in the scaffold density and decrease in the water swelling ratio of alginate. HNTs improve the stability of alginate scaffolds against enzymatic degradation in PBS solution. Thermogravimetrica analysis (TGA) shows that HNTs can improve the thermal stability of the alginate. The mouse fibroblast cells display better attachment to the alginate/HNT composite than those to the pure alginate, suggesting the good cytocompatibility of the composite scaffolds. Alginate/HNT composite scaffolds exhibit great potential for applications in tissue engineering. - Highlights: • We fabricated HNTs reinforced alginate composite scaffolds for biomedical applications. • The hydrogen bond interactions between HNTs and alginate are confirmed. • HNTs can significantly enhance the mechanical properties of alginate scaffold. • The scaffolds exhibit a highly porous structure with interconnected pores. • HNTs can improve the cell attachment and proliferation on alginate

  2. In vitro evaluation of alginate/halloysite nanotube composite scaffolds for tissue engineering

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Mingxian [Department of Materials Science and Engineering, Jinan University, Guangzhou 510632 (China); Dai, Libing [Guangzhou Institute of Traumatic Surgery, Guangzhou Red Cross Hospital Medical College, Jinan University, Guangzhou 510220 (China); Shi, Huizhe; Xiong, Sheng [Institute of Biomedicine, National Engineering Research Center of Genetic Medicine, Jinan University, Guangzhou 510632 (China); Zhou, Changren, E-mail: tcrz9@jnu.edu.cn [Department of Materials Science and Engineering, Jinan University, Guangzhou 510632 (China)

    2015-04-01

    In this study, a series of alginate/halloysite nanotube (HNTs) composite scaffolds were prepared by solution-mixing and freeze-drying method. HNTs are incorporated into alginate to improve both the mechanical and cell-attachment properties of the scaffolds. The interfacial interactions between alginate and HNTs were confirmed by the atomic force microscope (AFM), transmission electron microscope (TEM) and FTIR spectroscopy. The mechanical, morphological, and physico-chemical properties of the composite scaffolds were investigated. The composite scaffolds exhibit significant enhancement in compressive strength and compressive modulus compared with pure alginate scaffold both in dry and wet states. A well-interconnected porous structure with size in the range of 100–200 μm and over 96% porosity is found in the composite scaffolds. X-ray diffraction (XRD) result shows that HNTs are uniformly dispersed and partly oriented in the composite scaffolds. The incorporation of HNTs leads to increase in the scaffold density and decrease in the water swelling ratio of alginate. HNTs improve the stability of alginate scaffolds against enzymatic degradation in PBS solution. Thermogravimetrica analysis (TGA) shows that HNTs can improve the thermal stability of the alginate. The mouse fibroblast cells display better attachment to the alginate/HNT composite than those to the pure alginate, suggesting the good cytocompatibility of the composite scaffolds. Alginate/HNT composite scaffolds exhibit great potential for applications in tissue engineering. - Highlights: • We fabricated HNTs reinforced alginate composite scaffolds for biomedical applications. • The hydrogen bond interactions between HNTs and alginate are confirmed. • HNTs can significantly enhance the mechanical properties of alginate scaffold. • The scaffolds exhibit a highly porous structure with interconnected pores. • HNTs can improve the cell attachment and proliferation on alginate.

  3. the potential of alginic acid and polygal for soil stabilization

    African Journals Online (AJOL)

    user

    1985-09-01

    Sep 1, 1985 ... pertaining to the structure and physico-chemical properties of alginic acid, polygal and other polysaccharide types are available elsewhere [21]. 3. LABORATORY INVESTIGATIONS. Kaolinite was chosen to represent the lower bound of clay reactivity while montmorillonite represented the upper bounds.

  4. In vitro propagation of Acacia hybrid through alginate-encapsulated ...

    African Journals Online (AJOL)

    Seed collected from Acacia hybrid trees yields highly variable and poorly performing offspring and are not commonly used in regeneration. The present study described the incapsulation of Acacia hybrid shoots and axillary buds in the calcium alginate gel. The aim of the study was to evaluate the germination of the buds in ...

  5. Magnetic alginate microparticles for purification of .alpha.-amylases

    Czech Academy of Sciences Publication Activity Database

    Šafaříková, Miroslava; Roy, I.; Gupta, M. N.; Šafařík, Ivo

    2003-01-01

    Roč. 105, - (2003), s. 255-260 ISSN 0168-1656 R&D Projects: GA MŠk OC 523.80; GA AV ČR IBS6087204 Institutional research plan: CEZ:AV0Z6087904 Keywords : alginate * ferrofluid * amalyses Subject RIV: CE - Biochemistry Impact factor: 2.543, year: 2003

  6. Comparison Of The Dimensional Stability Of Alginate Impressions ...

    African Journals Online (AJOL)

    Methodology: Alginate impressions of a master model of truncated metal cones were made and disinfected with 1% sodium hypochlorite constituted from 3.5% household bleach using the spray and immersion technique for 10;20 and 30 minutes. Impressions were cast in dental stone and the linear dimensional differences ...

  7. Removal of Uranium from Aqueous Solution by Alginate Beads

    Directory of Open Access Journals (Sweden)

    Jing Yu

    2017-04-01

    Full Text Available The adsorption of uranium (VI by calcium alginate beads was examined by batch experiments. The effects of environmental conditions on U (VI adsorption were studied, including contact time, pH, initial concentration of U (VI, and temperature. The alginate beads were characterized by using scanning electron microscopy, transmission electron microscopy, X-ray photoelectron spectroscopy, and Fourier transform infrared spectroscopy. Fourier transform infrared spectra indicated that hydroxyl and alkoxy groups are present at the surface of the beads. The experimental results showed that the adsorption of U (VI by alginate beads was strongly dependent on pH, the adsorption increased at pH 3∼7, then decreased at pH 7∼9. The adsorption reached equilibrium within 2 minutes. The adsorption kinetics of U (VI onto alginate beads can be described by a pseudo first-order kinetic model. The adsorption isotherm can be described by the Redlich-Peterson model, and the maximum adsorption capacity was 237.15 mg/g. The sorption process is spontaneous and has an exothermic reaction.

  8. Bioinspired preparation of alginate nanoparticles using microbubble bursting.

    Science.gov (United States)

    Elsayed, Mohamed; Huang, Jie; Edirisinghe, Mohan

    2015-01-01

    Nanoparticles are considered to be one of the most advanced tools for drug delivery applications. In this research, alginate (a model hydrophilic polymer) nanoparticles 80 to 200 nm in diameter were obtained using microbubble bursting. The natural process of bubble bursting occurs through a number of stages, which consequently produce nano- and microsized droplets via two main production mechanisms, bubble shell disintegration and a jetting process. In this study, nano-sized droplets/particles were obtained by promoting the disintegrating mechanism and suppressing (limiting) the formation of larger microparticles resulting from the jetting mechanism. A T-junction microfluidic device was used to prepare alginate microbubbles with different sizes in a well-controlled manner. The size of the bubbles was varied by controlling two processing parameters, the solution flow rate and the bubbling pressure. Crucially, the bubble size was found to be the determining factor for inducing (or limiting) the bubble shell disintegration mechanism and the size needed to promote this process was influenced by the properties of the solution used for preparing the bubbles, particularly the viscosity. The size of alginate nanoparticles produced via the disintegration mechanism was found to be directly proportional to the viscosity of the alginate solution. Copyright © 2014 Elsevier B.V. All rights reserved.

  9. Continuous removal and recovery of lead by alginate beads, free ...

    African Journals Online (AJOL)

    This study examines the possibility of using Chlorella vulgaris cells in repeated lead adsorption/desorption cycles. Alginate beads and immobilized with algal cells were more effective and suitable than free cells. Consistently high lead removal (>90%) and recovery (about 100%) were achieved. Lead adsorption was mainly ...

  10. Development of Alginate/Chitosan Microparticles for Dust Mite ...

    African Journals Online (AJOL)

    Erah

    prepare a microparticulate system (coated with sodium alginate ... drop-wise addition of 10 ml TPP solution containing dust ... setup used the applied voltage for 18 kV. (Protek® DC ... Morphology and particle size distribution. Morphological ...

  11. Alginate Sulfate-Nanocellulose Bioinks for Cartilage Bioprinting Applications.

    Science.gov (United States)

    Müller, Michael; Öztürk, Ece; Arlov, Øystein; Gatenholm, Paul; Zenobi-Wong, Marcy

    2017-01-01

    One of the challenges of bioprinting is to identify bioinks which support cell growth, tissue maturation, and ultimately the formation of functional grafts for use in regenerative medicine. The influence of this new biofabrication technology on biology of living cells, however, is still being evaluated. Recently we have identified a mitogenic hydrogel system based on alginate sulfate which potently supports chondrocyte phenotype, but is not printable due to its rheological properties (no yield point). To convert alginate sulfate to a printable bioink, it was combined with nanocellulose, which has been shown to possess very good printability. The alginate sulfate/nanocellulose ink showed good printing properties and the non-printed bioink material promoted cell spreading, proliferation, and collagen II synthesis by the encapsulated cells. When the bioink was printed, the biological performance of the cells was highly dependent on the nozzle geometry. Cell spreading properties were maintained with the lowest extrusion pressure and shear stress. However, extruding the alginate sulfate/nanocellulose bioink and chondrocytes significantly compromised cell proliferation, particularly when using small diameter nozzles and valves.

  12. The Pseudomonas Quinolone Signal (PQS)

    DEFF Research Database (Denmark)

    Sams, Thomas; Baker, Ysobel; Hodgkinson, James

    2015-01-01

    Pseudomonas aeruginosa is an opportunistichuman pathogen that routinely appears near the top ofpublic health threat lists worldwide. P. aeruginosa causes in-fections by secreting a wealth of exceptionally active exo-products, leading to tissue damage. The synthesis of manyof these virulence factors...

  13. Secretory IgA as a diagnostic tool for Pseudomonas aeruginosa respiratory colonization

    DEFF Research Database (Denmark)

    Aanaes, Kasper; Johansen, Helle Krogh; Poulsen, Steen Seier

    2012-01-01

    BACKGROUND: Pseudomonas aeruginosa sinusitis may be the focus for intermittent lung colonization in patients with cystic fibrosis (CF). The sinusitis may induce elevated IgA levels in nasal secretion and saliva against P. aeruginosa. METHODS: 120 CF patients chronically infected, intermittently...... colonized or without P. aeruginosa in the lungs participated in this cross-sectional study. IgA and IgG against P. aeruginosa sonicate and alginate were measured in nasal secretions, saliva, and in serum by ELISA. RESULTS: The intermittently colonized patients had significantly higher IgA levels in nasal...... secretions and saliva than those without P. aeruginosa in the lungs, indicating that P. aeruginosa sinusitis may precede intermittent colonization and chronic infection of the lungs. CONCLUSIONS: Specific IgA against P. aeruginosa in nasal secretions and saliva can contribute to differentiation between...

  14. Colistin-Tobramycin Combinations Are Superior to Monotherapy Concerning the Killing of Biofilm Pseudomonas aeruginosa

    DEFF Research Database (Denmark)

    Herrmann, G.; Yang, Liang; Wu, H.

    2010-01-01

    Background. Antibiotic combination therapy might be more efficient than single antibiotics to combat Pseudomonas aeruginosa biofilms in the airways of patients with cystic fibrosis. We tested the ability of colistin sulphatetobramycin combinations and single antibiotics to kill P. aeruginosa...... biofilms. Methods. P. aeruginosa biofilms were generated in vitro and in rat lungs. In a pilot study, 5 patients with cystic fibrosis inhaled colistin and then tobramycin for 4 weeks. The changes in P. aeruginosa counts and lung function were assessed before and after therapy. Results. Antibiotic...... combination therapy significantly reduced the number of P. aeruginosa cells in P. aeruginosa biofilm models in vitro. When rats were challenged with 1 x 10(7) cfu of P. aeruginosa, which was embedded in alginate beads, mortality rates, lung pathologic findings, and bacterial colony-forming unit counts were...

  15. Expression, purification, crystallization and preliminary X-ray analysis of Pseudomonas aeruginosa AlgX

    Energy Technology Data Exchange (ETDEWEB)

    Weadge, J.T.; Robinson, H.; Yip, P. P.; Arnett, K.; Tipton, P. A.; Howell, P. L.

    2010-05-01

    AlgX is a periplasmic protein required for the production of the exopolysaccharide alginate in Pseudomonas sp. and Azotobacter vinelandii. AlgX has been overexpressed and purified and diffraction-quality crystals have been grown using iterative seeding and the hanging-drop vapor-diffusion method. The crystals grew as flat plates with unit-cell parameters a = 46.4, b = 120.6, c = 86.9 {angstrom}, {beta} = 95.7{sup o}. The crystals exhibited the symmetry of space group P2{sub 1} and diffracted to a minimum d-spacing of 2.1 {angstrom}. On the basis of the Matthews coefficient (V{sub M} = 2.25 {angstrom}{sup 3} Da{sup -1}), two molecules were estimated to be present in the asymmetric unit.

  16. Binding and leakage of barium in alginate microbeads.

    Science.gov (United States)

    Mørch, Yrr A; Qi, Meirigeng; Gundersen, Per Ole M; Formo, Kjetil; Lacik, Igor; Skjåk-Braek, Gudmund; Oberholzer, Jose; Strand, Berit L

    2012-11-01

    Microbeads of alginate crosslinked with Ca(2+) and/or Ba(2+) are popular matrices in cell-based therapy. The aim of this study was to quantify the binding of barium in alginate microbeads and its leakage under in vitro and accumulation under in vivo conditions. Low concentrations of barium (1 mM) in combination with calcium (50 mM) and high concentrations of barium (20 mM) in gelling solutions were used for preparation of microbeads made of high-G and high-M alginates. High-G microbeads accumulated barium from gelling solution and contained higher concentrations of divalent ions for both low- and high-Ba exposure compared with high-G microbeads exposed to calcium solely and to high-M microbeads for all gelling conditions. Although most of the unbound divalent ions were removed during the wash and culture steps, leakage of barium was still detected during storage. Barium accumulation in blood and femur bone of mice implanted with high-G beads was found to be dose-dependent. Estimated barium leakage relevant to transplantation to diabetic patients with islets in alginate microbeads showed that the leakage was 2.5 times lower than the tolerable intake value given by WHO for high-G microbeads made using low barium concentration. The similar estimate gave 1.5 times higher than is the tolerable intake value for the high-G microbeads made using high barium concentration. To reduce the risk of barium accumulation that may be of safety concern, the microbeads made of high-G alginate gelled with a combination of calcium and low concentration of barium ions is recommended for islet transplantation. Copyright © 2012 Wiley Periodicals, Inc.

  17. Survey of Bacterial and Fungal Contaminations in Iranian Alginate, Foreign Alginate and Speedex Used for Impression in Dentistry

    Directory of Open Access Journals (Sweden)

    Abbas Falah Tafti

    2012-02-01

    Full Text Available Background and Aims: Since impression materials usually contact with saliva, blood, and oral soft tissues, their microbial contamination are harmful in immunocompromised patients. The aim of the present study was to determine the bacterial and fungal contamination in common impression materials. Materials and Methods: In current lab trial study, 5 different samples from each 4 impression materials were homogenized in 1 ml Tween 80 and then 100µl of each sample were cultured onto blood agar, EMB, or sabouraud dextrose agar. Bacterial and fungal cultures were incubated at 37º C and 30º C, respectively. The isolated bacterial and fungal colonies were enumerated and identified using specific diagnostic media and tests. Data were analyzed using Kruskal-Wallis test. Results: Totally 75% of samples had one or several bacterial contaminations. Iranian alginate and Speedex (putty were the most contaminated samples. On the other hand, Speedex (light body and foreign alginate showed lower contamination. Species of Micrococcus, Staphylococcus, Bacilluses, Corynebacteria, gram negative Citrobacter, Actinomycetes and Neisseria were isolated from the analyzed impression materials. Aspergillus, Penicillium, Alternaria, Cladosporium and Sepdonium were the fungi isolated from impression materials. Statistical significant difference was shown between bacterial contamination of Iranian and foreign alginates (P=0.001. There was no statistical significant differences between the bacterial and fungal isolated colonies (CFU/gr of 4 tested impression materials (P=0.21. Conclusion: Several opportunistic bacteria and fungi were isolated from impression materials especially from Iranian alginate and Speedex putty which indicated their contamination.

  18. Experimental chronic Pseudomonas aeruginosa lung infection in rats. Non-specific stimulation with LPS reduces lethality as efficiently as specific immunization

    DEFF Research Database (Denmark)

    Lange, K H; Hougen, H P; Høiby, N

    1995-01-01

    In a rat model of chronic Pseudomonas aeruginosa lung infection mimicking cystic fibrosis, we investigated the possibility of preventing chronic lung inflammation or decreasing the progression of the infection. We compared the lethality, pathology, bacterial clearance, and immunogenicity after...... with either E. coli LPS or P. aeruginosa sonicate. Four and two weeks prior to challenge other rats were vaccinated with either E. coli LPS or P. aeruginosa sonicate. Controls did not receive any stimulation or vaccination. The lethality after challenge was lower in rats stimulated with E. coli LPS (p = 0...... but not to prevent the chronic P. aeruginosa lung infection and inflammation caused by alginate-embedded bacteria....

  19. Plant-associated fluorescent Pseudomonas from red lateritic soil: Beneficial characteristics and their impact on lettuce growth.

    Science.gov (United States)

    Maroniche, Guillermo A; Rubio, Esteban J; Consiglio, Adrián; Perticari, Alejandro

    2016-11-25

    Fluorescent Pseudomonas are ubiquitous soil bacteria that usually establish mutualistic associations with plants, promoting their growth and health by several mechanisms. This makes them interesting candidates for the development of crop bio-inoculants. In this work, we isolated phosphate-solubilizing fluorescent Pseudomonas from the rhizosphere and inner tissues of different plant species growing in red soil from Misiones, Argentina. Seven isolates displaying strong phosphate solubilization were selected for further studies. Molecular identification by rpoD genotyping indicated that they belong to different species within the P. fluorescens and P. putida phylogenetic groups. Screening for in vitro traits such as phosphate solubilization, growth regulators synthesis or degradation, motility and antagonism against phytopathogens or other bacteria, revealed a unique profile of characteristics for each strain. Their plant growth-promoting potential was assayed using lettuce as a model for inoculation under controlled and greenhouse conditions. Five of the strains increased the growth of lettuce plants. Overall, the strongest lettuce growth promoter under both conditions was strain ZME4, isolated from inner tissues of maize. No clear association between lettuce growth promotion and in vitro beneficial traits was detected. In conclusion, several phosphate solubilizing pseudomonads from red soil were isolated that display a rich array of plant growth promotion traits, thus showing a potential for the development of new inoculants.

  20. Alginate: Current Use and Future Perspectives in Pharmaceutical and Biomedical Applications

    Directory of Open Access Journals (Sweden)

    Marta Szekalska

    2016-01-01

    Full Text Available Over the last decades, alginates, natural multifunctional polymers, have increasingly drawn attention as attractive compounds in the biomedical and pharmaceutical fields due to their unique physicochemical properties and versatile biological activities. The focus of the paper is to describe biological and pharmacological activity of alginates and to discuss the present use and future possibilities of alginates as a tool in drug formulation. The recent technological advancements with using alginates, issues related to alginates suitability as matrix for three-dimensional tissue cultures, adjuvants of antibiotics, and antiviral agents in cell transplantation in diabetes or neurodegenerative diseases treatment, and an update on the antimicrobial and antiviral therapy of the alginate based drugs are also highlighted.

  1. Evaluation of fibroblasts adhesion and proliferation on alginate-gelatin crosslinked hydrogel.

    Directory of Open Access Journals (Sweden)

    Bapi Sarker

    Full Text Available Due to the relatively poor cell-material interaction of alginate hydrogel, alginate-gelatin crosslinked (ADA-GEL hydrogel was synthesized through covalent crosslinking of alginate di-aldehyde (ADA with gelatin that supported cell attachment, spreading and proliferation. This study highlights the evaluation of the physico-chemical properties of synthesized ADA-GEL hydrogels of different compositions compared to alginate in the form of films. Moreover, in vitro cell-material interaction on ADA-GEL hydrogels of different compositions compared to alginate was investigated by using normal human dermal fibroblasts. Viability, attachment, spreading and proliferation of fibroblasts were significantly increased on ADA-GEL hydrogels compared to alginate. Moreover, in vitro cytocompatibility of ADA-GEL hydrogels was found to be increased with increasing gelatin content. These findings indicate that ADA-GEL hydrogel is a promising material for the biomedical applications in tissue-engineering and regeneration.

  2. Synthesis of alginate oligomers by gamma irradiation and to investigate its antioxidant and prebiotic activity

    International Nuclear Information System (INIS)

    Bhoir, S.A.; Chawla, S.P.

    2016-01-01

    Alginate oligomers formed by alginate lyase have been reported to possess antioxidant activity as well as prebiotic activity. Hence, utility of gamma radiation to depolymerise alginate in its aqueous solution was investigated and its antioxidant and prebiotic activities were screened. 1% aqueous solution of sodium alginate was subjected to gamma irradiation and it's reducing power and ability to scavenge DPPH". and O_2"."."-, chelate iron and prevent heat induced β-carotene bleaching was determined. Prebiotic activity was determined by using alginate oligomers to promote prebiotic activity of Lactobacillus plantarum against E coli. Gamma radiation induced depolymerisation of alginate resulted in formation of oligomers with antioxidant and prebiotic activity. These polymers are potential candidates for utilization as natural preservatives and functional foods

  3. Variable Copy Number, Intra-Genomic Heterogeneities and Lateral Transfers of the 16S rRNA Gene in Pseudomonas

    Science.gov (United States)

    Bodilis, Josselin; Nsigue-Meilo, Sandrine; Besaury, Ludovic; Quillet, Laurent

    2012-01-01

    Even though the 16S rRNA gene is the most commonly used taxonomic marker in microbial ecology, its poor resolution is still not fully understood at the intra-genus level. In this work, the number of rRNA gene operons, intra-genomic heterogeneities and lateral transfers were investigated at a fine-scale resolution, throughout the Pseudomonas genus. In addition to nineteen sequenced Pseudomonas strains, we determined the 16S rRNA copy number in four other Pseudomonas strains by Southern hybridization and Pulsed-Field Gel Electrophoresis, and studied the intra-genomic heterogeneities by Denaturing Gradient Gel Electrophoresis and sequencing. Although the variable copy number (from four to seven) seems to be correlated with the evolutionary distance, some close strains in the P. fluorescens lineage showed a different number of 16S rRNA genes, whereas all the strains in the P. aeruginosa lineage displayed the same number of genes (four copies). Further study of the intra-genomic heterogeneities revealed that most of the Pseudomonas strains (15 out of 19 strains) had at least two different 16S rRNA alleles. A great difference (5 or 19 nucleotides, essentially grouped near the V1 hypervariable region) was observed only in two sequenced strains. In one of our strains studied (MFY30 strain), we found a difference of 12 nucleotides (grouped in the V3 hypervariable region) between copies of the 16S rRNA gene. Finally, occurrence of partial lateral transfers of the 16S rRNA gene was further investigated in 1803 full-length sequences of Pseudomonas available in the databases. Remarkably, we found that the two most variable regions (the V1 and V3 hypervariable regions) had probably been laterally transferred from another evolutionary distant Pseudomonas strain for at least 48.3 and 41.6% of the 16S rRNA sequences, respectively. In conclusion, we strongly recommend removing these regions of the 16S rRNA gene during the intra-genus diversity studies. PMID:22545126

  4. Bacterial Feeders, the Nematode Caenorhabditis elegans and the Flagellate Cercomonas longicauda, have different Effects on Outcome of Competition among the Pseudomonas Biocontrol Strains CHA0 and DSS73

    DEFF Research Database (Denmark)

    Pedersen, Annette; Nybroe, Ole; Winding, Anne

    2009-01-01

    How bacterial feeding fauna affects colonization and survival of bacteria in soil is not well understood, which constrains the applicability of bacterial inoculants in agriculture. This study aimed to unravel how food quality of bacteria and bacterial feeders with different feeding habits (the......50090 or one of two biocontrol strains P. fluorescens CHA0 or Pseudomonas sp. DSS73) or combinations of two bacterial strains. DSM50090 is a suitable food bacterium, DSS73 is of intermediate food quality, and CHA0 is inedible to the bacterial feeders. Bacterial and protozoan cell numbers were measured...... predation pressure. Hence, the results suggested that the outcome of competition among bacteria depended on their ability to cope with the prevailing bacterial predator....

  5. Effect of sodium alginate coating incorporated with nisin, Cinnamomum zeylanicum, and rosemary essential oils on microbial quality of chicken meat and fate of Listeria monocytogenes during refrigeration.

    Science.gov (United States)

    Raeisi, Mojtaba; Tabaraei, Alijan; Hashemi, Mohammad; Behnampour, Nasser

    2016-12-05

    The present study was conducted to preserve the microbial quality of chicken meat fillets during storage time by using sodium alginate active coating solutions incorporated with different natural antimicrobials including nisin, Cinnamomum zeylanicum (cinnamon), and rosemary essential oils (EOs) which were added individually and in combination. The samples were stored in refrigeration condition for 15days and were analyzed for total viable count, Enterobacteriaceae count, lactic acid bacteria count, Pseudomonas spp. count, psychrotrophic count, and yeast and mold count, as well as fate of inoculated Listeria monocytogenes at 3-day intervals. Results indicated that values of tested microbial indicators in all samples increased during storage. Antimicrobial agents, when used in combination, had stronger effect in preserving the microbial quality of chicken meat samples rather than their individual use and the strongest effect was observed in samples coated with alginate solution containing both cinnamon and rosemary EOs (CEO+REO). However, all treatments significantly inhibited microbial growth when compared to the control (Ppreservatives is recommended in meat products especially in chicken meats. Copyright © 2016. Published by Elsevier B.V.

  6. Surface characteristics determining the cell compatibility of ionically cross-linked alginate gels

    International Nuclear Information System (INIS)

    Machida-Sano, Ikuko; Hirakawa, Makoto; Matsumoto, Hiroki; Kamada, Mitsuki; Ogawa, Sakito; Satoh, Nao; Namiki, Hideo

    2014-01-01

    In this study we investigated differences in the characteristics determining the suitability of five types of ion (Fe 3+ , Al 3+ , Ca 2+ , Ba 2+ and Sr 2+ )-cross-linked alginate films as culture substrates for cells. Human dermal fibroblasts were cultured on each alginate film to examine the cell affinity of the alginates. Since cell behavior on the surface of a material is dependent on the proteins adsorbed to it, we investigated the protein adsorption ability and surface features (wettability, morphology and charge) related to the protein adsorption abilities of alginate films. We observed that ferric, aluminum and barium ion-cross-linked alginate films supported better cell growth and adsorbed higher amounts of serum proteins than other types. Surface wettability analysis demonstrated that ferric and aluminum ion-cross-linked alginates had moderate hydrophilic surfaces, while other types showed highly hydrophilic surfaces. The roughness was exhibited only on barium ion-cross-linked alginate surface. Surface charge measurements revealed that alginate films had negatively charged surfaces, and showed little difference among the five types of gel. These results indicate that the critical factors of ionically cross-linked alginate films determining the protein adsorption ability required for their cell compatibility may be surface wettability and morphology. (paper)

  7. Determination of bound and unbound water in dental alginate irreversible hydrocolloid by nuclear magnetic resonance spectroscopy.

    Science.gov (United States)

    Fellows, C M; Thomas, G A

    2009-04-01

    Alginate materials are considered unsuitable for precise fixed prosthetic rehabilitation due to their tendency to undergo spontaneous syneresis. Commercial alginate impression materials were investigated using Nuclear Magnetic Resonance (NMR) Spectroscopy to probe the relation between changes in the microscopic water environment and dimensional change to obtain a better understanding of spontaneous syneresis. NMR was used to measure the spin-lattice relaxation times (T(1)) of (1)H nuclei in water in alginate matrices to characterize changes in gel structure over time. These results were related to the dimensional stabilities of the alginate impression materials, their chemical compositions, and the Moisture Sorption Isotherms (MSI) obtained by incubation at fixed relative humidities. The rate of change of T(1) with time was found to be a better predictor of dimensional stability than MSI. The greatest dimensional stability for the alginate powders investigated was associated with a high filler:alginate ratio and a high Ca:Na ratio. Nuclear magnetic resonance spectroscopy may used to measure changes in alginate impression materials under conditions where no dimensional change can be observed directly. Changes occurred rapidly even at 100% humidity, suggesting the dimensional stability of alginate impression materials is partially independent of the rate of dehydration. The results may open a way to formulate alginate impression materials more suitable for precise fabrication of dental prostheses.

  8. Effects of disinfecting alginate impressions on the scratch hardness of stone models.

    Science.gov (United States)

    Hiraguchi, Hisako; Nakagawa, Hisami; Wakashima, Mitsuru; Miyanaga, Kohichi; Saigo, Masataka; Nishiyama, Minoru

    2006-03-01

    This study investigated the effects of disinfecting alginate impressions on the scratch depth of resultant stone models. Eleven brands of alginate impression material and two disinfectants, 1% sodium hypochlorite and 2% glutaraldehyde, were used. Impressions were immersed in disinfectant solutions or stored in sealed bags after spraying with disinfectants, and then poured with a type V dental stone. The scratch depth of the stone model obtained from disinfected impression was measured. The storage of alginate impressions after spraying with disinfectants did not increase the scratch depth of resultant stone models. However, the effect of immersion in disinfectants on scratch depth varied with the brand of the alginate impression material.

  9. In vivo evaluation of EPO-secreting cells immobilized in different alginate-PLL microcapsules.

    Science.gov (United States)

    Ponce, S; Orive, G; Hernández, R M; Gascón, A R; Canals, J M; Muñoz, M T; Pedraz, J L

    2006-11-01

    Alginates are the most employed biomaterials for cell encapsulation due to their abundance, easy gelling properties and apparent biocompatibility. However, as natural polymers different impurities including endotoxins, proteins and polyphenols can be found in their composition. Several purification protocols as well as different batteries of assays to prove the biocompatibility of the alginates in vitro have been recently developed. However, little is known about how the use of alginates with different purity grade may affect the host immune response after their implantation in vivo. The present paper investigates the long-term functionality and biocompatibility of murine erythropoietin (EPO) secreting C2C12 cells entrapped in microcapsules elaborated with alginates with different properties (purity, composition and viscosity). Results showed that independently of the alginate type employed, the animals presented elevated hematocrit levels until day 130, remaining at values between 70-87%. However, histological analysis of the explanted devices showed higher overgrowth around non-biomedical grade alginate microcapsules which could be directly related with higher impurity content of this type of alginate. Although EPO delivery may be limited by the formation of a fibrotic layer around non-biomedical grade alginate microcapsules, the high EPO secretion of the encapsulated cells together with the pharmacodynamic behaviour and the angiogenic and immune-modulatory properties of EPO result in no direct correlation between the biocompatibility of the alginate and the therapeutic response obtained.

  10. Clinical and laboratory studies of the antacid and raft-forming properties of Rennie alginate suspension.

    Science.gov (United States)

    Tytgat, G N; Simoneau, G

    2006-03-15

    Acid pockets at the gastro-oesophageal junction escape buffering from meals in the stomach. Combining high-dose antacid with alginate may therefore be of benefit in gastro-oesophageal reflux disease. To characterize the antacid and raft-forming properties of Rennie alginate suspension (containing high-dose antacid and alginate; Bayer Consumer Care, Bladel, the Netherlands). The in vitro acid-neutralizing capacity of Rennie algniate was compared with Gaviscon (Reckitt Benckiser, Slough, UK) by pH-recorded HCl titration. Alginate raft weight formed in vitro at different pH was used to evaluate the pH dependency of raft formation with each product. A double-blind, placebo-controlled, randomized crossover study also compared the antacid activity of Rennie alginate vs. placebo in vivo using continuous intragastric pH monitoring in 12 healthy fasting volunteers. Compared with Gaviscon, Rennie alginate had a higher acid-neutralizing capacity, greater maximum pH and longer duration of antacid activity in vitro. However, the two products produced comparable alginate rafts at each pH evaluated. In vivo, Rennie alginate provided rapid, effective and long-lasting acid neutralization, with an onset of action of <5 min, and duration of action of almost 90 min. The dual mode of action of Rennie alginate offers an effective treatment option for mild symptomatic gastro-oesophageal reflux disease particularly considering recent findings regarding 'acid pockets'.

  11. Drug release, preclinical and clinical pharmacokinetics relationships of alginate pellets prepared by melt technology.

    Science.gov (United States)

    Bose, Anirbandeep; Harjoh, Nurulaini; Pal, Tapan Kumar; Dan, Shubhasis; Wong, Tin Wui

    2016-01-01

    Alginate pellets prepared by the aqueous agglomeration technique experience fast drug dissolution due to the porous pre-formed calcium alginate microstructure. This study investigated in vitro drug release, preclinical and clinical pharmacokinetics relationships of intestinal-specific calcium acetate-alginate pellets against calcium-free and calcium carbonate-alginate pellets. Alginate pellets were prepared by solvent-free melt pelletization instead of aqueous agglomeration technique using chlorpheniramine maleate as model drug. A fast in situ calcium acetate dissolution in pellets resulted in rapid pellet breakup, soluble Ca(2+) crosslinking of alginate fragments and drug dissolution retardation at pH 1.2, which were not found in other pellet types. The preclinical drug absorption rate was lower with calcium acetate loaded than calcium-free alginate pellets. In human subjects, however, the extent and the rate of drug absorption were higher from calcium acetate-loaded pellets than calcium-free alginate pellets. The fine, dispersible and weakly gastric mucoadhesive calcium alginate pellets underwent fast human gastrointestinal transit. They released the drug at a greater rate than calcium-free pellets in the intestine, thereby promoting drug bioavailability. Calcium acetate was required as a disintegrant more than as a crosslinking agent clinically to promote pellet fragmentation, fast gastrointestinal transit and drug release in intestinal medium, and intestinal-specific drug bioavailability.

  12. Characteristics of Immobilized Urease on Grafted Alginate Bead Systems

    Directory of Open Access Journals (Sweden)

    Enas N. Danial

    2015-04-01

    Full Text Available This study evaluated the biological importance of immobilized urease enzyme over the free urease. The support material used for urease immobilization was alginate. Generally, the immobilization of urease in alginate gel showed a marked increase in Km and Vmax. However, the immobilized urease showed higher thermal stability than that of free enzyme. The rate of thermal inactivation of the immobilized enzyme decreased due to entrapment in gel matrix. Also, the activity of the immobilized urease was more stable in retention than that of the free enzyme during the storage in solution, although the activity of the immobilized enzyme was lower in comparison with the free enzyme. A stable immobilized system and long storage life are convenient for applications that would not be feasible with a soluble enzyme system. These results highlighted the technical and biochemical benefits of immobilized urease over the free enzyme.

  13. 3D-Printable Bioactivated Nanocellulose-Alginate Hydrogels.

    Science.gov (United States)

    Leppiniemi, Jenni; Lahtinen, Panu; Paajanen, Antti; Mahlberg, Riitta; Metsä-Kortelainen, Sini; Pinomaa, Tatu; Pajari, Heikki; Vikholm-Lundin, Inger; Pursula, Pekka; Hytönen, Vesa P

    2017-07-05

    We describe herein a nanocellulose-alginate hydrogel suitable for 3D printing. The composition of the hydrogel was optimized based on material characterization methods and 3D printing experiments, and its behavior during the printing process was studied using computational fluid dynamics simulations. The hydrogel was biofunctionalized by the covalent coupling of an enhanced avidin protein to the cellulose nanofibrils. Ionic cross-linking of the hydrogel using calcium ions improved the performance of the material. The resulting hydrogel is suitable for 3D printing, its mechanical properties indicate good tissue compatibility, and the hydrogel absorbs water in moist conditions, suggesting potential in applications such as wound dressings. The biofunctionalization potential was shown by attaching a biotinylated fluorescent protein and a biotinylated fluorescent small molecule via avidin and monitoring the material using confocal microscopy. The 3D-printable bioactivated nanocellulose-alginate hydrogel offers a platform for the development of biomedical devices, wearable sensors, and drug-releasing materials.

  14. Amplified fragment length polymorphism fingerprinting of Pseudomonas strains from a poultry processing plant.

    Science.gov (United States)

    Geornaras, I; Kunene, N F; von Holy, A; Hastings, J W

    1999-09-01

    Molecular typing has been used previously to identify and trace dissemination of pathogenic and spoilage bacteria associated with food processing. Amplified fragment length polymorphism (AFLP) is a novel DNA fingerprinting technique which is considered highly reproducible and has high discriminatory power. This technique was used to fingerprint 88 Pseudomonas fluorescens and Pseudomonas putida strains that were previously isolated from plate counts of carcasses at six processing stages and various equipment surfaces and environmental sources of a poultry abattoir. Clustering of the AFLP patterns revealed a high level of diversity among the strains. Six clusters (clusters I through VI) were delineated at an arbitrary Dice coefficient level of 0.65; clusters III (31 strains) and IV (28 strains) were the largest clusters. More than one-half (52.3%) of the strains obtained from carcass samples, which may have represented the resident carcass population, grouped together in cluster III. By contrast, 43.2% of the strains from most of the equipment surfaces and environmental sources grouped together in cluster IV. In most cases, the clusters in which carcass strains from processing stages grouped corresponded to the clusters in which strains from the associated equipment surfaces and/or environmental sources were found. This provided evidence that there was cross-contamination between carcasses and the abattoir environment at the DNA level. The AFLP data also showed that strains were being disseminated from the beginning to the end of the poultry processing operation, since many strains associated with carcasses at the packaging stage were members of the same clusters as strains obtained from carcasses after the defeathering stage.

  15. Hydrocarbonoclastic bacteria of the genus Pseudomonas in Samanea saman (Jacq. Merr. rhizosphere

    Directory of Open Access Journals (Sweden)

    Juliana Coromoto Mayz

    2017-01-01

    Full Text Available The research goal includes isolation, characterization and identification of Pseudomonas species existing in the rhizosphere of a legume present (colonizing or survivor in a savanna soil polluted by an oil spill, in order to explain the support of the growth of this leguminous plant through the reduction of the toxicity of spilled oil (hydrocarbonoclastic effects. The site is located at Amana del Tamarindo village entrance, Monagas State, Venezuela (9 ° 38' 52 "N, 63 ° 7' 20" E, 46 masl. An area of 50 m2 was sampled.  In concordance to the descriptions, keys, and comparison with the UOJ Herbarium exsiccatae, the legume collected was identified as Samanea saman (Jacq. Merr., which belongs to the Fabaceae family. The results of the biochemical characterization and the production of pyocyanine and fluorescein pigments allowed identifying 10 isolates as P. fluorescens, 5 as P. putida and 5 as P. aeruginosa. Samanea saman is recommended for re-vegetation of the contaminated area.

  16. Effect of Application of Pseudomonas fluorescent Strains on Yield and Yield Components of Rapeseed Cultivars

    Directory of Open Access Journals (Sweden)

    R Najafi

    2015-09-01

    Full Text Available Plant growth promoting rhizobacteria has been identified as an alternative to chemical fertilizer to enhance plant growth and yield directly and indirectly. Use of rhizosphere free living bacteria is one of the methods for crop production and leads to improvement of resources absorption. In order to study of yield, yield components and radiation use efficiency, under application of PGPR condition, an experiment was carried out in 2008 growing season at Agriculture and natural resources research station of Mashhad. The cultivars selected from three rapeseed species belong to Brassica napus, Brassica rapa and Brassica juncea (landrace, BP.18، Goldrush، Parkland، Hyola330، Hyola401. Experimental factorial design was randomized in complete block with three replications. Treatments included six varieties of Rapeseed and inoculations were four levels as non–inoculation, inoculation with P. fluorescens169, P. putida108 and use then together. Results showed that strains of fluorescent pseudomonas bacteria had greatest effects on yield and yield components cultivars. A significant difference in the number of pods per plant and 1000 seed weight observed. The cultivars were different in all treats except 1000 seed weight. Overall results indicated that application of growth stimulating bacteria in combination with different cultivars, had a positive effect growth, yield characteristics of plant varieties of rapeseed plants.

  17. Ancient acquisition of "alginate utilization loci" by human gut microbiota.

    Science.gov (United States)

    Mathieu, Sophie; Touvrey-Loiodice, Mélanie; Poulet, Laurent; Drouillard, Sophie; Vincentelli, Renaud; Henrissat, Bernard; Skjåk-Bræk, Gudmund; Helbert, William

    2018-05-23

    In bacteria from the phylum Bacteroidetes, the genes coding for enzymes involved in polysaccharide degradation are often colocalized and coregulated in so-called "polysaccharide utilization loci" (PULs). PULs dedicated to the degradation of marine polysaccharides (e.g. laminaran, ulvan, alginate and porphyran) have been characterized in marine bacteria. Interestingly, the gut microbiome of Japanese individuals acquired, by lateral transfer from marine bacteria, the genes involved in the breakdown of porphyran, the cell wall polysaccharide of the red seaweed used in maki. Sequence similarity analyses predict that the human gut microbiome also encodes enzymes for the degradation of alginate, the main cell wall polysaccharide of brown algae. We undertook the functional characterization of diverse polysaccharide lyases from family PL17, frequently found in marine bacteria as well as those of human gut bacteria. We demonstrate here that this family is polyspecific. Our phylogenetic analysis of family PL17 reveals that all alginate lyases, which have all the same specificity and mode of action, cluster together in a very distinct subfamily. The alginate lyases found in human gut bacteria group together in a single clade which is rooted deeply in the PL17 tree. These enzymes were found in PULs containing PL6 enzymes, which also clustered together in the phylogenetic tree of PL6. Together, biochemical and bioinformatics analyses suggest that acquisition of this system appears ancient and, because only traces of two successful transfers were detected upon inspection of PL6 and PL17 families, the pace of acquisition of marine polysaccharide degradation system is probably very slow.

  18. Determining the complex modulus of alginate irreversible hydrocolloid dental material.

    Science.gov (United States)

    King, Shalinie; See, Howard; Thomas, Graham; Swain, Michael

    2008-11-01

    The aim of the study is to investigate the visco-elastic response of an alginate irreversible hydrocolloid dental impression material during setting. A novel squeeze film Micro-Fourier Rheometer (MFR, GBC Scientific Equipment, Australia) was used to determine the complex modulus of an alginate irreversible hydrocolloid dental impression material (Algident, ISO 1563 Class A Type 1, Dentalfarm Australia Pty. Ltd.) during setting after mixing. Data was collected every 30s for 10 min in one study and every 10 min for a total of 60 min in another study. A high level of repeatability was observed. The results indicate that the MFR is capable of recording the complex shear modulus of alginate irreversible hydrocolloid for 60 min from the start of mixing and to simultaneously report the changing visco-elastic parameters at all frequencies between 1 Hz and 100 Hz. The storage modulus shows a dramatic increase to 370% of its starting value after 6 min and then reduces to 55% after 60 min. The loss modulus increases to a maximum of 175% of its starting value after 10 min and then reduces to 94% after 60 min. The MFR enables the changes in the complex modulus through the complete setting process to be followed. It is anticipated this approach may provide a better method to compare the visco-elastic properties of impression materials and assist with identification of optimum types for different clinical requirements. The high stiffness of the instrument and the use of band-limited pseudo-random noise as the input signal are the main advantages of this technique over conventional rheometers for determining the changes in alginate visco-elasticity.

  19. Characterization of alginate-brushite in-situ hydrogel composites

    Energy Technology Data Exchange (ETDEWEB)

    Dabiri, Seyed Mohammad Hossein [Department of Informatics, Bioengineering, Robotics, and System Engineering, University of Genoa, Genoa (Italy); Lagazzo, Alberto; Barberis, Fabrizio [Department of Civil, Chemical and Environmental Engineering, University of Genoa, Genoa (Italy); Farokhi, Mehdi [National Cell Bank of Iran, Pasteur Institute of Iran, Tehran (Iran, Islamic Republic of); Finochio, Elisabetta [Department of Civil, Chemical and Environmental Engineering, University of Genoa, Genoa (Italy); Pastorino, Laura [Department of Informatics, Bioengineering, Robotics, and System Engineering, University of Genoa, Genoa (Italy)

    2016-10-01

    In the present study alginate-brushite composite hydrogels were in-situ synthetized and characterized with respect to preparation parameters. Specifically, the influence of initial pH value and initial concentration of phosphate precursor on the in-situ fabrication of the composite hydrogel were taken into account. The composite hydrogels were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), thermogravimetric (TGA, DTG) and differential thermal analysis (DTA). Finally, the cell viability tests were carried out (MTT) over the incubation time period of 3, 7, and 14 days. The results revealed that the formation and the crystalline stability of brushite were highly dependent on the initial pH value. It was shown that as the pH reached to the value of 6, characteristics peaks of brushite appeared in the FTIR spectra. Besides, the XRD and thermal analysis results were in a good accordance with those of FTIR. In addition, the SEM images demonstrated that the plate like brushite was formed inside the alginate matrix. Also, a considerable impact of pH variation on the biocompatibility of samples was noticed so that the majority of samples especially those prepared in the acidic conditions were toxic. - Highlights: • Alginate-brushite hydrogel composites were obtained through an in-situ process • The brushite crystals started forming at pH value of 6 • The increase in the initial concentration of phosphate precursor resulted in more crystalline structure • Samples prepared at pH value of 8 had the most stable crystalline structure • Brushite crystals promoted the biocompatibility of alginate.

  20. Engineering Microvascularized 3D Tissue Using Alginate-Chitosan Microcapsules

    OpenAIRE

    Zhang, Wujie; Choi, Jung K.; He, Xiaoming

    2017-01-01

    Construction of vascularized tissues is one of the major challenges of tissue engineering. The goal of this study was to engineer 3D microvascular tissues by incorporating the HUVEC-CS cells with a collagen/alginate-chitosan (AC) microcapsule scaffold. In the presence of AC microcapsules, a 3D vascular-like network was clearly observable. The results indicated the importance of AC microcapsules in engineering microvascular tissues -- providing support and guiding alignment of HUVEC-CS cells. ...

  1. Growth and morphology of thermophilic dairy starters in alginate beads.

    Science.gov (United States)

    Lamboley, Laurence; St-Gelais, Daniel; Champagne, Claude P; Lamoureux, Maryse

    2003-06-01

    The aim of this research was to produce concentrated biomasses of thermophilic lactic starters using immobilized cell technology (ICT). Fermentations were carried out in milk using pH control with cells microentrapped in alginate beads. In the ICT fermentations, beads represented 17% of the weight. Some assays were carried out with free cells without pH control, in order to compare the ICT populations with those of classical starters. With Streptococcus thermophilus, overall populations in the fermentor were similar, but maximum bead population for (8.2 x 10(9) cfu/g beads) was 13 times higher than that obtained in a traditional starter (4.9 x 10(8) cfu/ml). For both Lactobacillus helveticus strains studied, immobilized-cell populations were about 3 x 10(9) cfu/g beads. Production of immobilized Lb. bulgaricus 210R strain was not possible, since no increases in viable counts occurred in beads. Therefore, production of concentrated cell suspension in alginate beads was more effective for S. thermophilus. Photomicrographs of cells in alginate beads demonstrated that, while the morphology of S. thermophilus remained unchanged during the ICT fermentation, immobilized cells of Lb. helveticus appeared wider. In addition, cells of Lb. bulgaricus were curved and elongated. These morphological changes would also impair the growth of immobilized lactobacilli.

  2. ENTRAPMENT OF FLUORESCENT E. COLI CELLS IN ALGINATE GEL

    Directory of Open Access Journals (Sweden)

    T. VINTILA

    2009-05-01

    Full Text Available By this experiment we will demonstrate the possibility to obtain genetically modified microbial strains that can be used as markers in different studies. The trait transferred in this study is the fluorescence in UV light expressed by a gene isolated from jellyfish. This gene was insered into a plasmid carrying ampiciline resistance and in the operon for arabinose fermentation. The plasmid was called pGLO. E coli HB101 K-12, ampicillin resistant colonies has been obtained. The colonies on the LB/amp/ara plate fluoresce green under UV light and the transformed colonies can grow on ampicillin. Transformation efficiency = 362 transformed colonies/ μg DNA. The cells where immobilized by entrapment in alginate gel to study the phenomenon involved in cells immobilization. After immobilization in alginate gel, 5x104 cells of E. coli pGLO / capsule and 1,4 x 105 cells of E. coli HB101/capsule has been found. Fluorescent microscopy revealed the presence of pGLO carrying cells into the capsules. After cultivation of alginate capsules containing E. coli in LB broth, and fluorescent microscopy of the capsule sections, several observations of the phenomenon involved in continuous fermentation using biocatalysts in has been made. These cells grow and migrate to the cortical part of the matrix where they are immobilized.

  3. IN VITRO EVALUATION OF FLUORIDE RELEASE OF JELTRATE® DENTAL ALGINATE

    Directory of Open Access Journals (Sweden)

    Matheus Melo Pithon

    2009-04-01

    Full Text Available Objective: To evaluate of fluoride release from Jeltrate alginate®. Materials and Methods: Four Trademarks of alginate were divided in four groups: conventional Jeltrate®, Plus Jeltrate®, Chromatic Jeltrate® and Chromatic Ortho Jeltrate®. The alginates were handled following the guidelines of the manufacturers. After this was followed by the construction of evidence bodies using silicone molds of the dimensions of 4 mm in diameter and 4mm in height. After take prey, the evidence bodies were removed from the molds and placed in container with 10 ml of ultra purified water, for 2 min. The fluoride release was measured by selective ion electrode connected to an analyzer of ions. Results: The Plus Jeltrate® showed a higher releasing fluoride 247.85 µg/cm2 followed by Chromatic Ortho Jeltrate® (217.83 µg/cm2, Chromatic Jeltrate ® (138.21 µg/cm2 and Jeltrate® (79.61 µg/cm2. Conclusion: Plus Jeltrate® had the best performance in releasing fluoride, followed by Chromatic Ortho Jeltrate®, Chromatic Jeltrate® and conventional Jeltrate®..

  4. Controlled fabrication of multi-core alginate microcapsules.

    Science.gov (United States)

    Eqbal, Md Danish; Gundabala, Venkat

    2017-12-01

    In this work, we present a robust microfluidic platform for controlled and complete on-chip generation of alginate microcapsules with single and double liquid cores. A combined Coflow and T-junction configuration implemented in a hybrid glass-PDMS (Polydimethylsiloxane) device is used for the generation of microcapsules with oil as liquid core. Frequency matching of oil-alginate double emulsion generation with that of aqueous Calcium chloride droplet generation allows for controlled merging of the two, resulting in reliable production of microcapsules. Confocal imaging of microcapsule cross-section reveals presence of intact liquid core. In the case of double core microcapsules, the two cores are well separated by alginate layer ensuring their long term stability. The current approach is expected to have advantages over existing techniques for liquid core microcapsule generation in terms of continuity of the process, control over core stability, and non-damage to cells when used for cell encapsulation applications. Copyright © 2017 Elsevier Inc. All rights reserved.

  5. Controlled antiseptic release by alginate polymer films and beads.

    Science.gov (United States)

    Liakos, Ioannis; Rizzello, Loris; Bayer, Ilker S; Pompa, Pier Paolo; Cingolani, Roberto; Athanassiou, Athanassia

    2013-01-30

    Biodegradable polymeric materials based on blending aqueous dispersions of natural polymer sodium alginate (NaAlg) and povidone iodine (PVPI) complex, which allow controlled antiseptic release, are presented. The developed materials are either free standing NaAlg films or Ca(2+)-cross-linked alginate beads, which properly combined with PVPI demonstrate antibacterial and antifungal activity, suitable for therapeutic applications, such as wound dressing. Glycerol was used as the plasticizing agent. Film morphology was studied by optical and atomic force microscopy. It was found that PVPI complex forms well dispersed circular micro-domains within the NaAlg matrix. The beads were fabricated by drop-wise immersion of NaAlg/PVPI/glycerol solutions into aqueous calcium chloride solutions to form calcium alginate beads encapsulating PVPI solution (CaAlg/PVPI). Controlled release of PVPI was possible when the composite films and beads were brought into direct contact with water or with moist media. Bactericidal and fungicidal properties of the materials were tested against Escherichia coli bacteria and Candida albicans fungi. The results indicated very efficient antibacterial and antifungal activity within 48 h. Controlled release of PVPI into open wounds is highly desired in clinical applications to avoid toxic doses of iodine absorption by the wound. A wide variety of applications are envisioned such as external and internal wound dressings with controlled antiseptic release, hygienic and protective packaging films for medical devices, and polymer beads as water disinfectants. Copyright © 2012 Elsevier Ltd. All rights reserved.

  6. Alginate Immobilization of Metabolic Enzymes (AIME) for High ...

    Science.gov (United States)

    Alginate Immobilization of Metabolic Enzymes (AIME) for High-Throughput Screening Assays DE DeGroot, RS Thomas, and SO SimmonsNational Center for Computational Toxicology, US EPA, Research Triangle Park, NC USAThe EPA’s ToxCast program utilizes a wide variety of high-throughput screening (HTS) assays to assess chemical perturbations of molecular and cellular endpoints. A key criticism of using HTS assays for toxicity assessment is the lack of xenobiotic metabolism (XM) which precludes both metabolic detoxification as well as bioactivation of chemicals tested in vitro thereby mischaracterizing the potential risk posed by these chemicals. To address this deficiency, we have developed an extracellular platform to retrofit existing HTS assays with XM activity. This platform utilizes the S9 fraction of liver homogenate encapsulated in an alginate gel network which reduces the cytotoxicity caused by direct addition of S9 to cells in culture. Alginate microspheres containing encapsulated human liver S9 were cross-linked to solid supports extending from a 96-well plate lid and were assayed using a pro-luciferin substrate specific for CYP3A4 (IPA). We demonstrate that S9 was successfully encapsulated and remained enzymatically active post-encapsulation with 5-10X the CYP3A4 activity as compared to 1 µg solubilized human liver S9. Ketoconazole, a known inhibitor of human CYP3A4, inhibited CYP3A4 activity in a concentration-dependent manner (IC50: 0.27 µM) and inhibiti

  7. From alginate impressions to digital virtual models: accuracy and reproducibility.

    Science.gov (United States)

    Dalstra, Michel; Melsen, Birte

    2009-03-01

    To compare the accuracy and reproducibility of measurements performed on digital virtual models with those taken on plaster casts from models poured immediately after the impression was taken, the 'gold standard', and from plaster models poured following a 3-5 day shipping procedure of the alginate impression. Direct comparison of two measuring techniques. The study was conducted at the Department of Orthodontics, School of Dentistry, University of Aarhus, Denmark in 2006/2007. Twelve randomly selected orthodontic graduate students with informed consent. Three sets of alginate impressions were taken from the participants within 1 hour. Plaster models were poured immediately from two of the sets, while the third set was kept in transit in the mail for 3-5 days. Upon return a plaster model was poured as well. Finally digital models were made from the plaster models. A number of measurements were performed on the plaster casts with a digital calliper and on the corresponding digital models using the virtual measuring tool of the accompanying software. Afterwards these measurements were compared statistically. No statistical differences were found between the three sets of plaster models. The intra- and inter-observer variability are smaller for the measurements performed on the digital models. Sending alginate impressions by mail does not affect the quality and accuracy of plaster casts poured from them afterwards. Virtual measurements performed on digital models display less variability than the corresponding measurements performed with a calliper on the actual models.

  8. Preparation of aminated chitosan/alginate scaffold containing halloysite nanotubes with improved cell attachment.

    Science.gov (United States)

    Amir Afshar, Hamideh; Ghaee, Azadeh

    2016-10-20

    The chemical nature of biomaterials play important role in cell attachment, proliferation and migration in tissue engineering. Chitosan and alginate are biodegradable and biocompatible polymers used as scaffolds for various medical and clinical applications. Amine groups of chitosan scaffolds play an important role in cell attachment and water adsorption but also associate with alginate carboxyl groups via electrostatic interactions and hydrogen bonding, consequently the activity of amine groups in the scaffold decreases. In this study, chitosan/alginate/halloysite nanotube (HNTs) composite scaffolds were prepared using a freeze-drying method. Amine treatment on the scaffold occurred through chemical methods, which in turn caused the hydroxyl groups to be replaced with carboxyl groups in chitosan and alginate, after which a reaction between ethylenediamine, 1-ethyl-3,(3-dimethylaminopropyl) carbodiimide (EDC) and scaffold triggered the amine groups to connect to the carboxyl groups of chitosan and alginate. The chemical structure, morphology and mechanical properties of the composite scaffolds were investigated by FTIR, CHNS, SEM/EDS and compression tests. The electrostatic attraction and hydrogen bonding between chitosan, alginate and halloysite was confirmed by FTIR spectroscopy. Chitosan/alginate/halloysite scaffolds exhibit significant enhancement in compressive strength compared with chitosan/alginate scaffolds. CHNS and EDS perfectly illustrate that amine groups were effectively introduced in the aminated scaffold. The growth and cell attachment of L929 cells as well as the cytotoxicity of the scaffolds were investigated by SEM and Alamar Blue (AB). The results indicated that the aminated chitosan/alginate/halloysite scaffold has better cell growth and cell adherence in comparison to that of chitosan/alginate/halloysite samples. Aminated chitosan/alginate/halloysite composite scaffolds exhibit great potential for applications in tissue engineering, ideally in

  9. Electrophoretic deposition of ZnO/alginate and ZnO-bioactive glass/alginate composite coatings for antimicrobial applications

    Energy Technology Data Exchange (ETDEWEB)

    Cordero-Arias, L.; Cabanas-Polo, S.; Goudouri, O.M. [Institute of Biomaterials, Department of Materials Science and Engineering, University of Erlangen-Nuremberg, Cauerstrasse 6, D-91058 Erlangen (Germany); Misra, S.K. [Materials Science and Engineering, Indian Institute of Technology Gandhinagar, Ahmedabad 382424 (India); Gilabert, J. [Institute of Ceramics Materials (ITC), University Jaume I, Avenida Vicent SosBaynat, 12006 Castellon (Spain); Valsami-Jones, E. [School of Geography, Earth and Environmental Sciences, University of Birmingham, Edgbaston, Birmingham B15 2TT (United Kingdom); Sanchez, E. [Institute of Ceramics Materials (ITC), University Jaume I, Avenida Vicent SosBaynat, 12006 Castellon (Spain); Virtanen, S. [Institute for Surface Science and Corrosion (LKO, WW4), Department of Materials Science and Engineering, University of Erlangen-Nuremberg, Martensstrasse 7, D-91058 Erlangen (Germany); Boccaccini, A.R., E-mail: aldo.boccaccini@ww.uni-erlangen.de [Institute of Biomaterials, Department of Materials Science and Engineering, University of Erlangen-Nuremberg, Cauerstrasse 6, D-91058 Erlangen (Germany)

    2015-10-01

    Two organic/inorganic composite coatings based on alginate, as organic matrix, and zinc oxide nanoparticles (n-ZnO) with and without bioactive glass (BG), as inorganic components, intended for biomedical applications, were developed by electrophoretic deposition (EPD). Different n-ZnO (1–10 g/L) and BG (1–1.5 g/L) contents were studied for a fixed alginate concentration (2 g/L). The presence of n-ZnO was confirmed to impart antibacterial properties to the coatings against gram-negative bacteria Escherichia coli, while the BG induced the formation of hydroxyapatite on coating surfaces thereby imparting bioactivity, making the coating suitable for bone replacement applications. Coating composition was analyzed by thermogravimetric analysis (TG), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and energy dispersive X-ray spectroscopy (EDS) analyses. Scanning electron microscopy (SEM) was employed to study both the surface and the cross section morphology of the coatings. Polarization curves of the coated substrates made in cell culture media at 37 °C confirmed the corrosion protection function of the novel organic/inorganic composite coatings. - Highlights: • Organic–inorganic nanocomposite coatings fabricated by electrophoretic deposition • nZnO and bioactive glass containing alginate coatings exhibit antibacterial effect. • Bioactive character and anticorrosion function of coatings demonstrated.

  10. A combined microcosm and mesocosm approach to examine factors affecting survival and mortality of Pseudomonas fluorescens Ag1 in seawater

    DEFF Research Database (Denmark)

    Ahl, Thomas; Christoffersen, K.; Riemann, B.

    1995-01-01

    .5‰). In microcosms with 0.2 µm-filtered seawater, addition of a carbon + nitrogen + phosphorus nutrient mixture was needed to induce proliferation of Ag1. In nutrient-amended microcosms the Ag1 population maintained viability, as determined by the direct viable counts method, at a level close to 100%. In natural...

  11. Elucidation of the Vibrio anguillarum genetic response to the potential fish probiont Pseudomonas fluorescens AH2, using RNA-arbitrarily primed PCR

    DEFF Research Database (Denmark)

    Holmstrøm, Kim; Gram, Lone

    2003-01-01

    or another V. anguillarum siderophore. On the pJM1 plasmid of V anguillarum serotype O1, a pseudogene designated open reading frame E (ORF E) that contains a frameshift mutation was previously identified. The gene homologous to vibE identified in this study, interestingly, also has significant homology...... to ORF E on the amino acid level and does not possess the frameshift mutation. Thus, the chromosomally encoded vibE homologue could fulfil the role of the inactive plasmid-encoded ORF E pseudogene. Addition of Fe3+ to the system eliminated the growth arrest, and the genes homologous to rpoS and vibE were...

  12. Structure Revision of N-Mercapto-4-formylcarbostyril Produced by Pseudomonas fluorescens G308 to 2-(2-Hydroxyphenyl)thiazole-4-carbaldehyde [aeruginaldehyde

    DEFF Research Database (Denmark)

    Guillemyn, Karel; Ballet, Steven; Ye, Lumeng

    2014-01-01

    , displays spectroscopic data identical with those of the alleged carbostyril derivative. In addition, the published H-1 and C-13 NMR data are in agreement with those calculated for aeruginaldehyde. We propose that aeruginaldehyde and aeruginol originate from the non-ribosomal peptide synthetase enzymes...

  13. Effects of spray-dried Pseudomonas fluorescens, strain CL145A (Zequanox®) on reproduction and early development of the fathead minnow (Pimephales promelas).

    Science.gov (United States)

    Waller, Diane L.; Luoma, James A.

    2016-01-01

    The biopesticide, Zequanox®, is registered for dreissenid mussel control in open water systems. Previous toxicity trials with nontarget organisms, including young-of-the year of several fish species and invertebrates, demonstrated selectivity of Zequanox for dreissenids. However, data are lacking on its safety to reproductive and early life stages of fish. The present study evaluated the effects of Zequanox on spawning and early life stages of the fathead minnow, Pimephales promelas, at the maximum approved concentration (100 mg Zequanox active ingredient /L) and exposure duration (8 h) for open water application. The results showed no significant effect of Zequanox on survival, condition, or cumulative egg deposition (21 d) in adult fathead minnow. Eggs (<24-h old) exposed to Zequanox developed to the eyed-stage at a similar rate to that of unexposed eggs. Additionally, Zequanox did not have a significant effect on survival and growth (90 d) of newly hatched fry (<24-h old). The results indicate that Zequanox treatment will not affect survival, spawning, and early life development of fathead minnows when applied at the recommended treatment regime.

  14. A New Twist to the Kirby-Bauer Antibiotic Susceptibility Test Activity?Increasing Antibiotic Sensitivity of Pseudomonas fluorescens through Thermal Stress

    OpenAIRE

    Gerbig, Donald G.; Engohang-Ndong, Jean; Aubihl, Heather

    2013-01-01

    Antibiotic sensitivity and the effect of temperature on microbial growth are two standard laboratory activities found in most microbial laboratory manuals. We have found a novel way to combine the two activities to demonstrate how temperature can influence antibiotic sensitivity using a standard incubator in instructional laboratory settings. This activity reinforces the important concepts of microbial growth and temperature along with Kirby-Bauer antibiotic susceptibility testing. We found t...

  15. Repeated batch and continuous degradation of chlorpyrifos by Pseudomonas putida.

    Science.gov (United States)

    Pradeep, Vijayalakshmi; Subbaiah, Usha Malavalli

    2015-01-01

    The present study was undertaken with the objective of studying repeated batch and continuous degradation of chlorpyrifos (O,O-diethyl O-3,5,6-trichloropyridin-2-yl phosphorothioate) using Ca-alginate immobilized cells of Pseudomonas putida isolated from an agricultural soil, and to study the genes and enzymes involved in degradation. The study was carried out to reduce the toxicity of chlorpyrifos by degrading it to less toxic metabolites. Long-term stability of pesticide degradation was studied during repeated batch degradation of chlorpyrifos, which was carried out over a period of 50 days. Immobilized cells were able to show 65% degradation of chlorpyrifos at the end of the 50th cycle with a cell leakage of 112 × 10(3) cfu mL(-1). During continuous treatment, 100% degradation was observed at 100 mL h(-1) flow rate with 2% chlorpyrifos, and with 10% concentration of chlorpyrifos 98% and 80% degradation was recorded at 20 mL h(-1) and 100 mL h(-1) flow rate respectively. The products of degradation detected by liquid chromatography-mass spectrometry analysis were 3,5,6-trichloro-2-pyridinol and chlorpyrifos oxon. Plasmid curing experiments with ethidium bromide indicated that genes responsible for the degradation of chlorpyrifos are present on the chromosome and not on the plasmid. The results of Polymerase chain reaction indicate that a ~890-bp product expected for mpd gene was present in Ps. putida. Enzymatic degradation studies indicated that the enzymes involved in the degradation of chlorpyrifos are membrane-bound. The study indicates that immobilized cells of Ps. putida have the potential to be used in bioremediation of water contaminated with chlorpyrifos.

  16. Pseudomonas-follikulitis efter badning i spabad

    DEFF Research Database (Denmark)

    Uldall Pallesen, Kristine Appel; Andersen, Klaus Ejner; Mørtz, Charlotte Gotthard

    2012-01-01

    . We describe a 23-year-old healthy woman who developed a pustular rash and general malaise after using a spa bath contaminated with Pseudomonas aeruginosa. Bacterial culture from a pustule confirmed Pseudomonas folliculitis and the patient was treated with ciprofloxacin with rapid good effect....

  17. Pseudomonas Septic Arthritis | Thanni | Nigerian Journal of ...

    African Journals Online (AJOL)

    BACKGROUND: Septic arthritis due to pseudomonas species is unusual and when it occurs, there is often an underlying cause like immune depression, intravenous drug abuse or a penetrating injury. PATIENT AND METHOD: We report a case of pseudomonas septic arthritis complicating cannulation of a leg vein following ...

  18. Mussel-inspired alginate gel promoting the osteogenic differentiation of mesenchymal stem cells and anti-infection

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Shiwen [Department of Mechanical Engineering, Faculty of Engineering and Department of Biochemistry & Genetics, Faculty of Medicine and Manitoba Institute of Child Health, The University of Manitoba, Winnipeg, Manitoba (Canada); Children Hospital Research Institute of Manitoba, Winnipeg (Canada); Sichuan University, Chengdu (China); Xu, Kaige; Darabi, Mohammad Ali [Children Hospital Research Institute of Manitoba, Winnipeg (Canada); Yuan, Quan [Sichuan University, Chengdu (China); Xing, Malcolm [Department of Mechanical Engineering, Faculty of Engineering and Department of Biochemistry & Genetics, Faculty of Medicine and Manitoba Institute of Child Health, The University of Manitoba, Winnipeg, Manitoba (Canada)

    2016-12-01

    Alginate hydrogels have been used in cell encapsulation for many years but a prevalent issue with pure alginates is that they are unable to provide enough bioactive properties to interact with mammalian cells. This paper discusses the modification of alginate with mussel-inspired dopamine for cell loading and anti-infection. Mouse bone marrow stem cells were immobilized into alginate and alginate-dopamine beads and fibers. Through live-dead and MTT assay, alginates modified by dopamine promoted cell viability and proliferation. In vitro cell differentiation results showed that such an alginate-dopamine gel can promote the osteogenic differentiation of mesenchymal stem cell after PCR and ALP assays. In addition to that, the adhesive prosperities of dopamine allowed for coating the surface of alginate-dopamine gel with silver nanoparticles, which provided the gel with significant antibacterial characteristics. Overall, these results demonstrate that a dopamine-modified alginate gel can be a great tool for cell encapsulation to promote cell proliferation and can be applied to bone regeneration, especially in contaminated bone defects. - Highlights: • Dopamine modified alginate bead and fiber promote cell viability and proliferation. • Alginate-dopamine gel promotes osteogenic differentiation of MSCs. • Dopamine reduced nanosilver for anti-infection. • Alginate-dopamine bead and fiber for delivery of mesenchymal stem cells (MSCs)

  19. Combination of sodium caseinate and succinylated alginate improved stability of high fat fish oil-in-water emulsions

    DEFF Research Database (Denmark)

    Yesiltas, Betül; Sørensen, Ann-Dorit Moltke; García Moreno, Pedro Jesús

    2018-01-01

    Sodium caseinate (CAS) and commercial sodium alginate (CA), long chain modified alginate (LCMA) or short chain modified alginate (SCMA) were used in combination for emulsifying and stabilizing high fat (50–70%) fish oil-in-water emulsions. Physical (creaming, droplet size, viscosity and protein...

  20. Mussel-inspired alginate gel promoting the osteogenic differentiation of mesenchymal stem cells and anti-infection

    International Nuclear Information System (INIS)

    Zhang, Shiwen; Xu, Kaige; Darabi, Mohammad Ali; Yuan, Quan; Xing, Malcolm

    2016-01-01

    Alginate hydrogels have been used in cell encapsulation for many years but a prevalent issue with pure alginates is that they are unable to provide enough bioactive properties to interact with mammalian cells. This paper discusses the modification of alginate with mussel-inspired dopamine for cell loading and anti-infection. Mouse bone marrow stem cells were immobilized into alginate and alginate-dopamine beads and fibers. Through live-dead and MTT assay, alginates modified by dopamine promoted cell viability and proliferation. In vitro cell differentiation results showed that such an alginate-dopamine gel can promote the osteogenic differentiation of mesenchymal stem cell after PCR and ALP assays. In addition to that, the adhesive prosperities of dopamine allowed for coating the surface of alginate-dopamine gel with silver nanoparticles, which provided the gel with significant antibacterial characteristics. Overall, these results demonstrate that a dopamine-modified alginate gel can be a great tool for cell encapsulation to promote cell proliferation and can be applied to bone regeneration, especially in contaminated bone defects. - Highlights: • Dopamine modified alginate bead and fiber promote cell viability and proliferation. • Alginate-dopamine gel promotes osteogenic differentiation of MSCs. • Dopamine reduced nanosilver for anti-infection. • Alginate-dopamine bead and fiber for delivery of mesenchymal stem cells (MSCs)

  1. Characterization of alginates from Ghanaian brown seaweeds: Sargassum spp. and Padina spp

    DEFF Research Database (Denmark)

    Rhein-Knudsen, Nanna; Ale, Marcel Tutor; Ajalloueian, Fatemeh

    2017-01-01

    Alginates of four locally harvested Ghanaian brown seaweeds from the Sargassum and Padina genus were assessed for their rheological and chemical characteristics. The seaweeds contained 16–30% by weight of alginate assessed as the sum of d-mannuronic acid (M) and l-guluronic acid (G). In compariso...

  2. Photocrosslinked alginate with hyaluronic acid hydrogels as vehicles for mesenchymal stem cell encapsulation and chondrogenesis.

    Science.gov (United States)

    Coates, Emily E; Riggin, Corinne N; Fisher, John P

    2013-07-01

    Ionic crosslinking of alginate via divalent cations allows for high viability of an encapsulated cell population, and is an effective biomaterial for supporting a spherical chondrocyte morphology. However, such crosslinking chemistry does not allow for injectable and stable hydrogels which are more appropriate for clinical applications. In this study, the addition of methacrylate groups to the alginate polymer chains was utilized so as to allow the free radical polymerization initiated by a photoinitiator during UV light exposure. This approach establishes covalent crosslinks between methacrylate groups instead of the ionic crosslinks formed by the calcium in unmodified alginate. Although this approach has been well described in the literature, there are currently no reports of stem cell differentiation and subsequent chondrocyte gene expression profiles in photocrosslinked alginate. In this study, we demonstrate the utility of photocrosslinked alginate hydrogels containing interpenetrating hyaluronic acid chains to support stem cell chondrogenesis. We report high cell viability and no statistical difference in metabolic activity between mesenchymal stem cells cultured in calcium crosslinked alginate and photocrosslinked alginate for up to 10 days of culture. Furthermore, chondrogenic gene markers are expressed throughout the study, and indicate robust differentiation up to the day 14 time point. At early time points, days 1 and 7, the addition of hyaluronic acid to the photocrosslinked scaffolds upregulates gene markers for both the chondrocyte and the superficial zone chondrocyte phenotype. Taken together, we show that photocrosslinked, injectable alginate shows significant potential as a delivery mechanism for cell-based cartilage repair therapies. Copyright © 2012 Wiley Periodicals, Inc.

  3. Alginate as a cell culture substrate for growth and differentiation of human retinal pigment epithelial cells.

    Science.gov (United States)

    Heidari, Razeih; Soheili, Zahra-Soheila; Samiei, Shahram; Ahmadieh, Hamid; Davari, Maliheh; Nazemroaya, Fatemeh; Bagheri, Abouzar; Deezagi, Abdolkhalegh

    2015-03-01

    The purpose of this study was to evaluate retinal pigment epithelium (RPE) cells' behavior in alginate beads that establish 3D environment for cellular growth and mimic extracellular matrix versus the conventional 2D monolayer culture. RPE cells were encapsulated in alginate beads by dripping alginate cell suspension into CaCl2 solution. Beads were suspended in three different media including Dulbecco's modified Eagle's medium (DMEM)/F12 alone, DMEM/F12 supplemented with 10 % fetal bovine serum (FBS), and DMEM/F12 supplemented with 30 % human amniotic fluid (HAF). RPE cells were cultivated on polystyrene under the same conditions as controls. Cell phenotype, cell proliferation, cell death, and MTT assay, immunocytochemistry, and real-time RT-PCR were performed to evaluate the effect of alginate on RPE cells characteristics and integrity. RPE cells can survive and proliferate in alginate matrixes. Immunocytochemistry analysis exhibited Nestin, RPE65, and cytokeratin expressions in a reasonable number of cultured cells in alginate beads. Real-time PCR data demonstrated high levels of Nestin, CHX10, RPE65, and tyrosinase gene expressions in RPE cells immobilized in alginate when compared to 2D monolayer culture systems. The results suggest that alginate can be used as a reliable scaffold for maintenance of RPE cells' integrity and in vitro propagation of human retinal progenitor cells for cell replacement therapies in retinal diseases.

  4. In vitro investigation of the integration depth of oral fluids and disinfectants into alginate impressions.

    Science.gov (United States)

    Surna, Rimas; Junevicius, Jonas; Rutkauskas, Evaldas

    2009-01-01

    The objective of this work is to prove that oral cavity fluids diffuse into alginate mass of impressions. In addition, the information is presented on the subject that disinfectants used for alginate impressions disinfection not only diffuse into alginate mass but penetrate deeper than oral cavity fluids. Three examination groups were formed for the research, the results of which evidenced how deeply oral cavity fluids and disinfectants 'Alpha Guard GF' and 'Orbis' could possibly diffuse into alginate impression material 'Kromopan 100'. In the first examination group ten impressions from the upper jaw dental arch and mucosa were taken, firstly colouring oral cavity fluids with a special colouring tablet MIRA-2-TON (Hager Werken). Cuts were randomly selected from impressions and scanned aiming to establish the depth of the coloured oral cavity fluid penetration. In the second and the third examination groups taken alginate impressions were accordingly soaked in 'Alpha Guard GF' and 'Orbis' with pigment and later randomly selected cuts were scanned in the same manner as in the first research group. RESULTS. The research results establish that coloured dental cavity fluids maximum diffuse into alginate impression is up to 540 microm with the presence of 95% of discolouring while disinfectants 'Alpha Guard GF' and 'Orbis' accordingly diffuse into alginate mass up to 710 microm and 870 microm with the presence of 95% of discolouring. CONCLUSIONS. The results obtained show that disinfectants using them according to the recommendations of a manufacturer, diffuse into alginate mass deeper than oral cavity fluids at the time of impressions taking.

  5. Evaluation of setting time and flow properties of self-synthesize alginate impressions

    Science.gov (United States)

    Halim, Calista; Cahyanto, Arief; Sriwidodo, Harsatiningsih, Zulia

    2018-02-01

    Alginate is an elastic hydrocolloid dental impression materials to obtain negative reproduction of oral mucosa such as to record soft-tissue and occlusal relationships. The aim of the present study was to synthesize alginate and to determine the setting time and flow properties. There were five groups of alginate consisted of fifty samples self-synthesize alginate and commercial alginate impression product. Fifty samples were divided according to two tests, each twenty-five samples for setting time and flow test. Setting time test was recorded in the s unit, meanwhile, flow test was recorded in the mm2 unit. The fastest setting time result was in the group three (148.8 s) and the latest was group fours). The highest flow test result was in the group three (69.70 mm2) and the lowest was group one (58.34 mm2). Results were analyzed statistically by one way ANOVA (α= 0.05), showed that there was a statistical significance of setting time while no statistical significance of flow properties between self-synthesize alginate and alginate impression product. In conclusion, the alginate impression was successfully self-synthesized and variation composition gives influence toward setting time and flow properties. The most resemble setting time of control group is group three. The most resemble flow of control group is group four.

  6. Clinical and laboratory studies of the antacid and raft-forming properties of Rennie alginate suspension

    NARCIS (Netherlands)

    Tytgat, G. N.; Simoneau, G.

    2006-01-01

    BACKGROUND: Acid pockets at the gastro-oesophageal junction escape buffering from meals in the stomach. Combining high-dose antacid with alginate may therefore be of benefit in gastro-oesophageal reflux disease. AIM: To characterize the antacid and raft-forming properties of Rennie alginate

  7. Synthesis of hydrogels of alginate for system controlled release of progesterone

    International Nuclear Information System (INIS)

    Abreu, Marlon de F.; Rodriguez, Ruben J.S.; Silva, Ester C.C. da; Barreto, Gabriela N.S.

    2015-01-01

    The chemical modifications of natural polymers like alginate, has allowed the development of new formulations for controlled release systems. In this work we report the synthesis of a derivative of the amidic alginate with alkyl chain. The polymer was characterized by spectroscopic techniques: Nuclear Magnetic Resonance and Fourier Transform Infrared. (author)

  8. Review: peripheral nerve regeneration using non-tubular alginate gel crosslinked with covalent bonds.

    Science.gov (United States)

    Hashimoto, Tadashi; Suzuki, Yoshihisa; Suzuki, Kyoko; Nakashima, Toshihide; Tanihara, Masao; Ide, Chizuka

    2005-06-01

    We have developed a nerve regeneration material consisting of alginate gel crosslinked with covalent bonds. in the first part of this study, we attempted to analyze nerve regeneration through alginate gel in the early stages within 2 weeks. in the second part, we tried to regenerate cat peripheral nerve by using alginate tubular or non-tubular nerve regeneration devices, and compared their efficacies. Four days after surgery, regenerating axons grew without Schwann cell investment through the partially degraded alginate gel, being in direct contact with the alginate without a basal lamina covering. One to 2 weeks after surgery, regenerating axons were surrounded by common Schwann cells, forming small bundles, with some axons at the periphery being partly in direct contact with alginate. At the distal stump, numerous Schwann cells had migrated into the alginate 8-14 days after surgery. Remarkable restorations of the 50-mm gap in cat sciatic nerve were obtained after a long term by using tubular or non-tubular nerve regeneration material consisting mainly of alginate gel. However, there was no significant difference between both groups at electrophysiological and morphological evaluation. Although, nowadays, nerve regeneration materials being marketed mostly have a tubular structure, our results suggest that the tubular structure is not indispensable for peripheral nerve regeneration.

  9. The effect of chitosan molecular weight on the properties of alginate ...

    African Journals Online (AJOL)

    Purpose: The aim of the present study was to investigate the effect of chitosan molecular weight on size, size distribution, release rate, mucoadhesive properties and electrostatic bonding of alginate/chitosan microparticles containing prednisolone. Methods: Three mucoadhesive alginate/chitosan microparticle formulations, ...

  10. Alginic Acid-Aided Dispersion of Carbon Nanotubes, Graphene, and Boron Nitride Nanomaterials for Microbial Toxicity Testing.

    Science.gov (United States)

    Wang, Ying; Mortimer, Monika; Chang, Chong Hyun; Holden, Patricia A

    2018-01-30

    Robust evaluation of potential environmental and health risks of carbonaceous and boron nitride nanomaterials (NMs) is imperative. However, significant agglomeration of pristine carbonaceous and boron nitride NMs due to strong van der Waals forces renders them not suitable for direct toxicity testing in aqueous media. Here, the natural polysaccharide alginic acid (AA) was used as a nontoxic, environmentally relevant dispersant with defined composition to disperse seven types of carbonaceous and boron nitride NMs, including multiwall carbon nanotubes, graphene, boron nitride nanotubes, and hexagonal boron nitride flakes, with various physicochemical characteristics. AA's biocompatibility was confirmed by examining AA effects on viability and growth of two model microorganisms (the protozoan Tetrahymena thermophila and the bacterium Pseudomonas aeruginosa ). Using 400 mg·L -1 AA, comparably stable NM (200 mg·L -1 ) stock dispersions were obtained by 30-min probe ultrasonication. AA non-covalently interacted with NM surfaces and improved the dispersibility of NMs in water. The dispersion stability varied with NM morphology and size rather than chemistry. The optimized dispersion protocol established here can facilitate preparing homogeneous NM dispersions for reliable exposures during microbial toxicity testing, contributing to improved reproducibility of toxicity results.

  11. Alginic Acid-Aided Dispersion of Carbon Nanotubes, Graphene, and Boron Nitride Nanomaterials for Microbial Toxicity Testing

    Directory of Open Access Journals (Sweden)

    Ying Wang

    2018-01-01

    Full Text Available Robust evaluation of potential environmental and health risks of carbonaceous and boron nitride nanomaterials (NMs is imperative. However, significant agglomeration of pristine carbonaceous and boron nitride NMs due to strong van der Waals forces renders them not suitable for direct toxicity testing in aqueous media. Here, the natural polysaccharide alginic acid (AA was used as a nontoxic, environmentally relevant dispersant with defined composition to disperse seven types of carbonaceous and boron nitride NMs, including multiwall carbon nanotubes, graphene, boron nitride nanotubes, and hexagonal boron nitride flakes, with various physicochemical characteristics. AA’s biocompatibility was confirmed by examining AA effects on viability and growth of two model microorganisms (the protozoan Tetrahymena thermophila and the bacterium Pseudomonas aeruginosa. Using 400 mg·L−1 AA, comparably stable NM (200 mg·L−1 stock dispersions were obtained by 30-min probe ultrasonication. AA non-covalently interacted with NM surfaces and improved the dispersibility of NMs in water. The dispersion stability varied with NM morphology and size rather than chemistry. The optimized dispersion protocol established here can facilitate preparing homogeneous NM dispersions for reliable exposures during microbial toxicity testing, contributing to improved reproducibility of toxicity results.

  12. Antifungal Effect of a Dental Tissue Conditioner Containing Nystatin-Loaded Alginate Microparticles.

    Science.gov (United States)

    Kim, Hyun-Jin; Son, Jun Sik; Kwon, Tae-Yub

    2018-02-01

    In this in vitro study, nystatin-alginate microparticles were successfully fabricated to control the release of nystatin from a commercial dental tissue conditioner. These nystatin-alginate microparticles were spherical and had a slightly rough surface. The microparticles incorporated into the tissue conditioner were distributed homogeneously throughout the tissue conditioner matrix. The incorporation of the microparticles did not deteriorate the mechanical properties of the original material. The agar diffusion test results showed that the tissue conditioner containing the microparticles had a good antifungal effect against Candida albicans. The nystatin-alginate microparticles efficiently controlled the release of nystatin from the tissue conditioner matrix over the experimental period of 14 days. Moreover, the nystatin-alginate microparticles incorporated in the tissue conditioner showed effective antifungal function even at lower concentrations of nystatin. The current study suggests that the tissue conditioner containing the nystatin-alginate microparticle carrier system has potential as an effective antifungal material.

  13. Synthesis and Characterization of Crosslinked Hydrogel Polyacrylamide (PAAM)-Co-Alginate Prepared by Gama Irradiation

    International Nuclear Information System (INIS)

    Erizal; Tita P; Dewi SP

    2008-01-01

    Crosslinked poly(acrylamide) (PAAM)-co-alginate hydrogels were prepared by gamma irradiation (γ-irradiation) and their conditions such as irradiation dose and alginate concentration were studied. PAAM-co-alginate was crosslinked to yield water sorption materials with various ability to absorb water (swelling) depending on the preparation conditions (e.g. γ-irradiation dosage>20 kGy) and alginate concentration (0.5 - 1 wt %). With an increase of γ-irradiation dosage and alginate concentration, the gels content and water absorption were increasing markedly. The swelling properties of hydrogel in urea and NaCl solution and the effect of temperature were also investigated. Intensity decreasing of functional goups of OH and NH 2 in the IR spectrum indicated that IPN (Interpenetreting Network) structure occured in the network of hydrogels. The ability of hydrogel to absorp and retain a large amount of water suggested their possible uses in health care and agriculture. (author)

  14. Calcium alginate dressings promote healing of split skin graft donor sites.

    LENUS (Irish Health Repository)

    O'Donoghue, J M

    2012-02-03

    A prospective controlled trial was carried out to assess the healing efficacy of calcium alginate and paraffin gauze on split skin graft donor sites. Thirty patients were randomised to the calcium alginate group and 21 to the paraffin gauze group. The donor sites were assessed at 10 days post harvesting to determine if they were completely healed (100%) or not. Twenty one of the 30 patients dressed with calcium alginate were completely healed at day 10, while only 7\\/21 in the paraffin gauze group were healed (p < 0.05). There were two infections in the study, both occurring in the alginate group while there was no difference in dressing slippage between the two groups. Calcium alginate dressings provide a significant improvement in healing split skin graft donor sites.

  15. Physical and chemical characterization of titanium-alginate samples for biomedical applications

    Energy Technology Data Exchange (ETDEWEB)

    Morani, L.M.; Ribeiro, A.A.; Oliveira, M.V. de; Dantas, F.M.L., E-mail: marize.varella@int.gov.b [Instituto Nacional de Tecnologia (INT), Rio de Janeiro, RJ (Brazil); Leao, M.H.M.R. [Universidade Federal do Rio de Janeiro (EQ/UFRJ), RJ (Brazil). Escola de Quimica

    2010-07-01

    The sol-gel technique combined with powder metallurgy may be an alternative to produce titanium parts for bioengineering, with the advantage of eliminating the powder compaction step, which may introduce defects. The present work introduces a system consisted of titanium powder and sodium alginate suspension, which undergoes reticulation in contact with a calcium salt solution, obtaining titanium/calcium alginate hydrogel with granule morphology. The characterization of the raw materials and granules of calcium alginate and titanium/calcium alginate was performed by x-ray fluorescence spectroscopy and thermogravimetric analysis. The granules topography was analyzed by scanning electron microscopy/EDS. Titanium and sodium alginate chemical composition were adequate for use as raw materials, showing that the methodology used is suitable for processing titanium samples for further consolidation by sintering, in order to produce titanium parts. (author)

  16. Genipin Cross-Linked Polymeric Alginate-Chitosan Microcapsules for Oral Delivery: In-Vitro Analysis

    Directory of Open Access Journals (Sweden)

    Hongmei Chen

    2009-01-01

    Full Text Available We have previously reported the preparation of the genipin cross-linked alginate-chitosan (GCAC microcapsules composed of an alginate core with a genipin cross-linked chitosan membrane. This paper is the further investigation on their structural and physical characteristics. Results showed that the GCAC microcapsules had a smooth and dense surface and a networked interior. Cross-linking by genipin substantially reduced swelling and physical disintegration of microcapsules induced by nongelling ions and calcium sequestrants. Strong resistance to mechanical shear forces and enzymatic degradation was observed. Furthermore, the GCAC membranes were permeable to bovine serum albumin and maintained a molecular weight cutoff at 70 KD, analogous to the widely studied alginate-chitosan, and alginate-poly-L-lysine-alginate microcapsules. The release features and the tolerance of the GCAC microcapsules in the stimulated gastrointestinal environment were also investigated. This GCAC microcapsule formulation offers significant potential as a delivery vehicle for many biomedical applications.

  17. Computational prediction of the Crc regulon identifies genus-wide and species-specific targets of catabolite repression control in Pseudomonas bacteria

    Directory of Open Access Journals (Sweden)

    O'Gara Fergal

    2010-11-01

    Full Text Available Abstract Background Catabolite repression control (CRC is an important global control system in Pseudomonas that fine tunes metabolism in order optimise growth and metabolism in a range of different environments. The mechanism of CRC in Pseudomonas spp. centres on the binding of a protein, Crc, to an A-rich motif on the 5' end of an mRNA resulting in translational down-regulation of target genes. Despite the identification of several Crc targets in Pseudomonas spp. the Crc regulon has remained largely unexplored. Results In order to predict direct targets of Crc, we used a bioinformatics approach based on detection of A-rich motifs near the initiation of translation of all protein-encoding genes in twelve fully sequenced Pseudomonas genomes. As expected, our data predict that genes related to the utilisation of less preferred nutrients, such as some carbohydrates, nitrogen sources and aromatic carbon compounds are targets of Crc. A general trend in this analysis is that the regulation of transporters is conserved across species whereas regulation of specific enzymatic steps or transcriptional activators are often conserved only within a species. Interestingly, some nucleoid associated proteins (NAPs such as HU and IHF are predicted to be regulated by Crc. This finding indicates a possible role of Crc in indirect control over a subset of genes that depend on the DNA bending properties of NAPs for expression or repression. Finally, some virulence traits such as alginate and rhamnolipid production also appear to be regulated by Crc, which links nutritional status cues with the regulation of virulence traits. Conclusions Catabolite repression control regulates a broad spectrum of genes in Pseudomonas. Some targets are genus-wide and are typically related to central metabolism, whereas other targets are species-specific, or even unique to particular strains. Further study of these novel targets will enhance our understanding of how Pseudomonas

  18. Computational prediction of the Crc regulon identifies genus-wide and species-specific targets of catabolite repression control in Pseudomonas bacteria.

    Science.gov (United States)

    Browne, Patrick; Barret, Matthieu; O'Gara, Fergal; Morrissey, John P

    2010-11-25

    Catabolite repression control (CRC) is an important global control system in Pseudomonas that fine tunes metabolism in order optimise growth and metabolism in a range of different environments. The mechanism of CRC in Pseudomonas spp. centres on the binding of a protein, Crc, to an A-rich motif on the 5' end of an mRNA resulting in translational down-regulation of target genes. Despite the identification of several Crc targets in Pseudomonas spp. the Crc regulon has remained largely unexplored. In order to predict direct targets of Crc, we used a bioinformatics approach based on detection of A-rich motifs near the initiation of translation of all protein-encoding genes in twelve fully sequenced Pseudomonas genomes. As expected, our data predict that genes related to the utilisation of less preferred nutrients, such as some carbohydrates, nitrogen sources and aromatic carbon compounds are targets of Crc. A general trend in this analysis is that the regulation of transporters is conserved across species whereas regulation of specific enzymatic steps or transcriptional activators are often conserved only within a species. Interestingly, some nucleoid associated proteins (NAPs) such as HU and IHF are predicted to be regulated by Crc. This finding indicates a possible role of Crc in indirect control over a subset of genes that depend on the DNA bending properties of NAPs for expression or repression. Finally, some virulence traits such as alginate and rhamnolipid production also appear to be regulated by Crc, which links nutritional status cues with the regulation of virulence traits. Catabolite repression control regulates a broad spectrum of genes in Pseudomonas. Some targets are genus-wide and are typically related to central metabolism, whereas other targets are species-specific, or even unique to particular strains. Further study of these novel targets will enhance our understanding of how Pseudomonas bacteria integrate nutritional status cues with the regulation

  19. Computational prediction of the Crc regulon identifies genus-wide and species-specific targets of catabolite repression control in Pseudomonas bacteria

    LENUS (Irish Health Repository)

    Browne, Patrick

    2010-11-25

    Abstract Background Catabolite repression control (CRC) is an important global control system in Pseudomonas that fine tunes metabolism in order optimise growth and metabolism in a range of different environments. The mechanism of CRC in Pseudomonas spp. centres on the binding of a protein, Crc, to an A-rich motif on the 5\\' end of an mRNA resulting in translational down-regulation of target genes. Despite the identification of several Crc targets in Pseudomonas spp. the Crc regulon has remained largely unexplored. Results In order to predict direct targets of Crc, we used a bioinformatics approach based on detection of A-rich motifs near the initiation of translation of all protein-encoding genes in twelve fully sequenced Pseudomonas genomes. As expected, our data predict that genes related to the utilisation of less preferred nutrients, such as some carbohydrates, nitrogen sources and aromatic carbon compounds are targets of Crc. A general trend in this analysis is that the regulation of transporters is conserved across species whereas regulation of specific enzymatic steps or transcriptional activators are often conserved only within a species. Interestingly, some nucleoid associated proteins (NAPs) such as HU and IHF are predicted to be regulated by Crc. This finding indicates a possible role of Crc in indirect control over a subset of genes that depend on the DNA bending properties of NAPs for expression or repression. Finally, some virulence traits such as alginate and rhamnolipid production also appear to be regulated by Crc, which links nutritional status cues with the regulation of virulence traits. Conclusions Catabolite repression control regulates a broad spectrum of genes in Pseudomonas. Some targets are genus-wide and are typically related to central metabolism, whereas other targets are species-specific, or even unique to particular strains. Further study of these novel targets will enhance our understanding of how Pseudomonas bacteria integrate

  20. Pyoverdine synthesis by the Mn(II-oxidizing bacterium Pseudomonas putida GB-1

    Directory of Open Access Journals (Sweden)

    Dorothy Lundquist Parker

    2014-05-01

    Full Text Available When iron-starved, the Mn(II-oxidizing bacteria Pseudomonas putida strains GB-1 and MnB1 produce pyoverdines (PVDGB-1 and PVDMnB1, siderophores that both influence iron uptake and inhibit manganese(II oxidation by these strains. To explore the properties and genetics of a PVD that can affect manganese oxidation, LC-MS/MS and various siderotyping techniques were used to identify the peptides of PVDGB-1 and PVDMnB1 as being (for both PVDs: chromophore-Asp-Lys-OHAsp-Ser-Gly-aThr-Lys-cOHOrn, resembling a structure previously reported for P. putida CFML 90-51, which does not oxidize Mn. All three strains also produced an azotobactin and a sulfonated PVD, each with the peptide sequence above, but with unknown regulatory or metabolic effects. Bioinformatic analysis of the sequenced genome of P. putida GB-1 suggested that a particular non-ribosomal peptide synthetase, coded by the operon PputGB1_4083-4086, could produce the peptide backbone of PVDGB-1. To verify this prediction, plasmid integration disruption of PputGB1_4083 was performed and the resulting mutant failed to produce detectable PVD. In silico analysis of the modules in PputGB1_4083-4086 predicted a peptide sequence of Asp-Lys-Asp-Ser-Ala-Thr-Lsy-Orn, which closely matches the peptide determined by MS/MS. To extend these studies to other organisms, various Mn(II-oxidizing and non-oxidizing isolates of P. putida, P. fluorescens, P. marincola, P. fluorescens-syringae group, P. mendocina-resinovorans group and P. stutzerii group were screened for PVD synthesis. The PVD producers (12 out of 16 tested strains were siderotyped and placed into four sets of differing PVD structures, some corresponding to previously characterized PVDs and some to novel PVDs. These results combined with previous studies suggested that the presence of OHAsp or the flexibility of the pyoverdine polypeptide may enable efficient binding of Mn(III.

  1. Pyoverdine synthesis by the Mn(II)-oxidizing bacterium Pseudomonas putida GB-1

    Science.gov (United States)

    Parker, Dorothy L.; Lee, Sung-Woo; Geszvain, Kati; Davis, Richard E.; Gruffaz, Christelle; Meyer, Jean-Marie; Torpey, Justin W.; Tebo, Bradley M.

    2014-01-01

    When iron-starved, the Mn(II)-oxidizing bacteria Pseudomonas putida strains GB-1 and MnB1 produce pyoverdines (PVDGB-1 and PVDMnB1), siderophores that both influence iron uptake and inhibit manganese(II) oxidation by these strains. To explore the properties and genetics of a PVD that can affect manganese oxidation, LC-MS/MS, and various siderotyping techniques were used to identify the peptides of PVDGB-1 and PVDMnB1 as being (for both PVDs): chromophore-Asp-Lys-OHAsp-Ser-Gly-aThr-Lys-cOHOrn, resembling a structure previously reported for P. putida CFML 90-51, which does not oxidize Mn. All three strains also produced an azotobactin and a sulfonated PVD, each with the peptide sequence above, but with unknown regulatory or metabolic effects. Bioinformatic analysis of the sequenced genome of P. putida GB-1 suggested that a particular non-ribosomal peptide synthetase (NRPS), coded by the operon PputGB1_4083-4086, could produce the peptide backbone of PVDGB-1. To verify this prediction, plasmid integration disruption of PputGB1_4083 was performed and the resulting mutant failed to produce detectable PVD. In silico analysis of the modules in PputGB1_4083-4086 predicted a peptide sequence of Asp-Lys-Asp-Ser-Ala-Thr-Lsy-Orn, which closely matches the peptide determined by MS/MS. To extend these studies to other organisms, various Mn(II)-oxidizing and non-oxidizing isolates of P. putida, P. fluorescens, P. marincola, P. fluorescens-syringae group, P. mendocina-resinovorans group, and P. stutzerii group were screened for PVD synthesis. The PVD producers (12 out of 16 tested strains) were siderotyped and placed into four sets of differing PVD structures, some corresponding to previously characterized PVDs and some to novel PVDs. These results combined with previous studies suggested that the presence of OHAsp or the flexibility of the pyoverdine polypeptide may enable efficient binding of Mn(III). PMID:24847318

  2. Two kinds of ketoprofen enteric gel beads (CA and CS-SA using biopolymer alginate

    Directory of Open Access Journals (Sweden)

    Bingchao Cheng

    2018-03-01

    Full Text Available To obtain expected rapid-release and sustained-release of ketoprofen gel beads, this paper adopted biopolymer alginate to prepare alginate beads and chitosan-alginate gel beads. Formulation factors were investigated and optimized by the single factor test. The release of ketoprofen from calcium alginate gel beads in pH 1.0 hydrochloric acid solution was less than 10% during 2 h, then in pH6.8 was about 95% during 45 min, which met the requirements of rapid-release preparations. However, the drug release of chitosan-alginate gel beads in pH1.0 was less than 5% during 2 h, then in pH6.8 was about 50% during 6 h and reached more than 95% during 12 h, which had a good sustained-release behavior. In addition, the release kinetics of keteprofen from the calcium alginate gel beads fitted well with the Korsmeyer–Peppas model and followed a case-II transport mechanism. However, the release of keteprofen from the chitosan-alginate gel beads exhibited a non-Fickian mechanism and based on the mixed mechanisms of diffusion and polymer relaxation from chitosan-alginate beads. In a word, alginate gel beads of ketoprofen were instant analgesic, while chitosan-alginate gel beads could control the release of ketoprofen during gastro-intestinal tract and prolong the drug's action time. Keywords: Gel beads, Enteric rapid-release, Enteric sustained-release, Ketoprofen

  3. Sustained release of verapamil hydrochloride from sodium alginate microcapsules.

    Science.gov (United States)

    Farhana, S Ayesha; Shantakumar, S M; Shyale, Somashekar; Shalam, Md; Narasu, Laxmi

    2010-04-01

    The objective of the present study was to develop sustained release microcapsules of verapamil hydrochloride (VH) using biodegradable polymers. For this purpose microcapsules embedded verapamil hydrochloride were prepared using sodium alginate alone and also by incorporating some co polymers like methyl cellulose (MC), sodium carboxy methyl cellulose (SCMC) , poly vinyl pyrollidone (PVP) and xanthan gum by employing complex emulsion method of microencapsulation. Microcapsules were prepared in various core: coat ratios to know the effect of polymer and co polymers on drug release. Overall ten formulations were prepared and evaluated for flow behaviour, sieve analysis, drug entrapment efficiency, in vitro dissolution studies, stability studies, including scanning electron microscopy and DSC. The resulting microcapsules were discrete, large, spherical and also free flowing. The drug content in all the batches of microcapsules was found to be uniform. The release was depended on core: coat ratio and nature of the polymers. FTIR analysis revealed chemical integrity between Verapamil hydrochloride (VH), sodium alginate and between the copolymers. Among the four copolymers used methyl cellulose retarded the drug release more than the other three, hence the same formulation was subjected for in vivo studies. The drug release from the microcapsules was found to be following non fickian diffusion. Mechanism of drug release was diffusion controlled first order kinetics. Drug diffusion co efficient and correlation co efficient were also assessed by using various mathematical models. In vivo result analysis of pharmacokinetic parameters revealed that t max of reference and test formulations were almost same. From the study it was concluded that, sustained release Verapamil hydro chloride microcapsules could be achieved with success using sodium alginate alone and also in combination with other biodegradable polymers.

  4. Immunization with Pseudomonas aeruginosa vaccines and adjuvant can modulate the type of inflammatory response subsequent to infection

    DEFF Research Database (Denmark)

    Johansen, H K; Espersen, F; Cryz, S J

    1994-01-01

    Pseudomonas aeruginosa is the predominant pathogen in patients with cystic fibrosis (CF). To study the possibility of preventing lung inflammation and decreasing the progression of the infection by vaccination, we have developed a rat model of chronic P. aeruginosa lung infection. Rats were......, 2.0 versus 2.1 to 2.8) pathologic abnormalities were less severe in the control rats injected with sterile saline than in the immunized rats and the IFA group. The more severe lung abnormalities observed in immunized rats could be due to the result of immune complex-mediated lung tissue damage...... an acute-type inflammation to a chronic-type inflammation dominated by mononuclear leukocytes and scattered granulomas. Cross-reacting antibodies were induced by the two alginate vaccines, and most immunized animals developed a significant (P

  5. Self-assembled chitosan-alginate polyplex nanoparticles containing temoporfin

    Czech Academy of Sciences Publication Activity Database

    Brezaniova, I.; Trousil, Jiří; Černochová, Zulfiya; Král, V.; Hrubý, Martin; Štěpánek, Petr; Šlouf, Miroslav

    2017-01-01

    Roč. 295, č. 8 (2017), s. 1259-1270 ISSN 0303-402X R&D Projects: GA MŠk(CZ) LQ1604; GA MŠk(CZ) ED1.1.00/02.0109; GA ČR(CZ) GA16-02870S; GA MZd(CZ) NV15-25781A; GA MŠk(CZ) LO1507 Institutional support: RVO:61389013 Keywords : chitosan * sodium alginate * temoporfin Subject RIV: CD - Macromolecular Chemistry OBOR OECD: Polymer science Impact factor: 1.723, year: 2016

  6. Preparation and Characterization of Keratin/Alginate Blend Microparticles

    OpenAIRE

    Srisuwan, Yaowalak; Srihanam, Prasong

    2018-01-01

    The water-in-oil (W/O) emulsification-diffusion method was used for construction of keratin (Ker), alginate (Alg), and Ker/Alg blend microparticles. The Ker, Alg, and Ker/Alg blend solutions were used as the water phase, while ethyl acetate was used as the oil phase. Firstly, different concentrations of Ker solution was used to find suitable content. 1.6% w/v Ker solution was blended with the same concentration of the Alg solution for further microparticle construction. Results from scanning ...

  7. Engineering Microvascularized 3D Tissue Using Alginate-Chitosan Microcapsules.

    Science.gov (United States)

    Zhang, Wujie; Choi, Jung K; He, Xiaoming

    2017-02-01

    Construction of vascularized tissues is one of the major challenges of tissue engineering. The goal of this study was to engineer 3D microvascular tissues by incorporating the HUVEC-CS cells with a collagen/alginate-chitosan (AC) microcapsule scaffold. In the presence of AC microcapsules, a 3D vascular-like network was clearly observable. The results indicated the importance of AC microcapsules in engineering microvascular tissues -- providing support and guiding alignment of HUVEC-CS cells. This approach provides an alternative and promising method for constructing vascularized tissues.

  8. Characterisation of bare and tannase-loaded calcium alginate beads by microscopic, thermogravimetric, FTIR and XRD analyses.

    Science.gov (United States)

    Larosa, Claudio; Salerno, Marco; de Lima, Juliana Silva; Merijs Meri, Remo; da Silva, Milena Fernandes; de Carvalho, Luiz Bezerra; Converti, Attilio

    2018-08-01

    Incorporating enzymes into calcium alginate beads is an effective method to immobilise them and to preserve, at the same time, their catalytic activity. Sodium alginate was mixed with Aspergillus ficuum tannase in aqueous solution, and tannase-loaded calcium alginate beads were prepared using a simple droplet-based microfluidic system. Extensive experimental analysis was carried out to characterise the samples. Microscopic imaging revealed morphological differences between the surfaces of bare alginate matrix and tannase-loaded alginate beads. Thermal analysis allowed assessing the hydration contents of alginate and revealed the presence of tannase entrapped in the loaded beads, which was confirmed by vibrational spectroscopy. X-ray diffraction allowed us to conclude that alginate of tannase-loaded beads is not crystalline, which would make them suitable as carriers for possible controlled release. Moreover, they could be used in food applications to improve tea quality or clarify juices. Copyright © 2018 Elsevier B.V. All rights reserved.

  9. Silver against Pseudomonas aeruginosa biofilms

    DEFF Research Database (Denmark)

    Bjarnsholt, Thomas; Kirketerp-Møller, K.; Kristiansen, S.

    2007-01-01

    bacteria in both the planktonic and biofilm modes of growth. The action of silver on mature in vitro biofilms of Pseudomonas aeruginosa, a primary pathogen of chronic infected wounds, was investigated. The results show that silver is very effective against mature biofilms of P. aeruginosa......, but that the silver concentration is important. A concentration of 5-10 ig/mL silver sulfadiazine eradicated the biofilm whereas a lower concentration (1 ig/mL) had no effect. The bactericidal concentration of silver required to eradicate the bacterial biofilm was 10-100 times higher than that used to eradicate...... planktonic bacteria. These observations strongly indicate that the concentration of silver in currently available wound dressings is much too low for treatment of chronic biofilm wounds. It is suggested that clinicians and manufacturers of the said wound dressings consider whether they are treating wounds...

  10. Pseudomona pseudomallei community acquired pneumonia

    International Nuclear Information System (INIS)

    Severiche, Diego

    1998-01-01

    This is the first published case report en Colombia about pseudomona pseudomallei community acquired pneumonia. This uncommon pathogen is from the epidemiological standpoint a very important one and medical community should be aware to look after it in those patients where no other etiological pathogen is recovered. A brief summary about epidemiology is showed, emphasizing those regions where it can be found. Likewise, comments about the differential diagnosis are important since it should be considered in those patients where tuberculosis is suspected. This is particularly representative for countries with high tuberculosis rates. Furthermore, a microbiological review is shown, emphasizing on isolation techniques, descriptions about therapeutics and other regarding treatment issues according international standards. Finally; a description about the clinical picture, laboratory findings, treatment and evolution of the case reported are shown for discussion

  11. Glyphosate catabolism by Pseudomonas sp

    International Nuclear Information System (INIS)

    Shinabarger, D.L.

    1986-01-01

    The pathway for the degradation of glyphosate (N-phosphonomethylglycine) by Pseudomonas sp. PG2982 has been determined using metabolic radiolabeling experiments. Radiorespirometry experiments utilizing [3- 14 C] glyphosate revealed that approximately 50-59% of the C3 carbon was oxidized to CO 2 . Fractionation of stationary phase cells labeled with [3- 14 C]glyphosate revealed that from 45-47% of the assimilated C3 carbon is distributed to proteins and that amino acids methionine and serine are highly labeled. The nucleic acid bases adenine and guanine received 90% of the C3 label that was incorporated into nucleic acids, and the only pyrimidine base labeled was thymine. Pulse labeling of PG2982 cells with [3- 14 C]glyphosate revealed that [3- 14 C]sarcosine is an intermediate in glyphosate degradation. Examination of crude extracts prepared from PG2982 cells revealed the presence of an enzyme that oxidizes sarcosine to glycine and formaldehyde. These results indicate that the first step in glyphosate degradation by PG2982 is cleavage of the carbon-phosphorus bond, resulting in the release of sarcosine and a phosphate group. The phosphate group is utilized as a source of phosphorus, and the sarcosine is degraded to glycine and formaldehyde. Phosphonate utilization by Pseudomonas sp. PG2982 was investigated. Each of the ten phosphonates tested were utilized as a sole source of phosphorus by PG2982. Representative compounds tested included alkylphosphonates, 1-amino-substituted alkylphosphonates, amino-terminal phosphonates, and an arylphosphonate. PG2982 cultures degraded phenylphosphonate to benzene and produced methane from methylphosphonate. The data indicate that PG2982 is capable of cleaving the carbon-phosphorus bond of several structurally different phosphonates

  12. The enhancement of chondrogenesis of ATDC5 cells in RGD-immobilized microcavitary alginate hydrogels.

    Science.gov (United States)

    Yao, Yongchang; Zeng, Lei; Huang, Yuyang

    2016-07-01

    In our previous work, we have developed an effective microcavitary alginate hydrogel for proliferation of chondrocytes and maintenance of chondrocytic phenotype. In present work, we investigated whether microcavitary alginate hydrogel could promote the chondrogenesis of progenitor cells. Moreover, we attempted to further optimize this system by incorporating synthetic Arg-Gly-Asp peptide. ATDC5 cells were seeded into microcavitary alginate hydrogel with or without Arg-Gly-Asp immobilization. Cell Counting Kit-8 and live/dead staining were conducted to analyze cell proliferation. Real-time polymerase chain reaction (RT-PCR), hematoxylin and eosin, and Toluidine blue O staining as well as Western blot assay was performed to evaluate the cartilaginous markers at transcriptional level and at protein level, respectively. The obtained data demonstrated that Arg-Gly-Asp-immobilized microcavitary alginate hydrogel was preferable to promote the cell proliferation. Also, Arg-Gly-Asp-immobilized microcavitary alginate hydrogel improved the expression of chondrocytic genes including Collagen II and Aggrecan when compared with microcavitary alginate hydrogel. The results suggested that microcavitary alginate hydrogel could promote the chondrogenesis. And Arg-Gly-Asp would be promising to ameliorate this culture system for cartilage tissue engineering. © The Author(s) 2016.

  13. Alginate hydrogel as a promising scaffold for dental-derived stem cells: an in vitro study.

    Science.gov (United States)

    Moshaverinia, Alireza; Chen, Chider; Akiyama, Kentaro; Ansari, Sahar; Xu, Xingtian; Chee, Winston W; Schricker, Scott R; Shi, Songtao

    2012-12-01

    The objectives of this study were to: (1) develop an injectable and biodegradable scaffold based on oxidized alginate microbeads encapsulating periodontal ligament (PDLSCs) and gingival mesenchymal stem cells (GMSCs); and (2) investigate the stem cell viability, and osteogenic differentiation of the stem cells in vitro. Stem cells were encapsulated using alginate hydrogel. The stem cell viability, proliferation and differentiation to adipogenic and osteogenic tissues were studied. To investigate the expression of both adipogenesis and ontogenesis related genes, the RNA was extracted and RT-PCR was performed. The degradation behavior of hydrogel based on oxidized sodium alginate with different degrees of oxidation was studied in PBS at 37 °C as a function of time by monitoring the changes in weight loss. The swelling kinetics of alginate hydrogel was also investigated. The results showed that alginate is a promising candidate as a non-toxic scaffold for PDLSCs and GMSCs. It also has the ability to direct the differentiation of these stem cells to osteogenic and adipogenic tissues as compared to the control group in vitro. The encapsulated stem cells remained viable in vitro and both osteo-differentiated and adipo-differentiated after 4 weeks of culturing in the induction media. It was found that the degradation profile and swelling kinetics of alginate hydrogel strongly depends on the degree of oxidation showing its tunable chemistry and degradation rate. These findings demonstrate for the first time that immobilization of PDLSCs and GMSCs in the alginate microspheres provides a promising strategy for bone tissue engineering.

  14. Accuracy and dimensional stability of extended-pour and conventional alginate impression materials.

    Science.gov (United States)

    Imbery, Terence A; Nehring, Joshua; Janus, Charles; Moon, Peter C

    2010-01-01

    The authors conducted a study to determine if two irreversible hydrocolloid impression materials (Cavex ColorChange, Cavex Holland BV, Haarlem, Netherlands; Jeltrate Plus Antimicrobial Dustless Alginate Impression Material, Dentsply Caulk, Milford, Del.) stored for five days were dimensionally accurate. The authors modified Ivorine teeth (Columbia Dentoform, Long Island City, N.Y.) on a Dentoform model (1560 series model, Columbia Dentoform) to allow measurements of tooth and arch width. They made impressions and generated casts immediately and at five additional times. They recorded tooth and arch widths on the casts and compared the measurements with those for the standard model. Compared with measurements for the model, the greatest measured difference in casts was 0.003 inches for Cavex ColorChange (extended-pour alginate) and 0.005 inches for Jeltrate Plus Antimicrobial Dustless Alginate Impression Material (conventional alginate). The percentage of dimensional change ranged from -0.496 to 0.161 percent for the extended-pour alginate and from -0.174 to 0.912 percent for the conventional alginate. Results of analysis of variance and paired t tests indicated that when generated immediately and at day 5, casts produced from both impression materials were not statistically different from the standard model (P alginate materials can produce accurate impressions at day 5 for diagnostic casts and for fabrication of acrylic appliances.

  15. In vitro evaluation of calcium alginate gels as matrix for iontophoresis electrodes.

    Science.gov (United States)

    Haida, Haruka; Ando, Shizuka; Ogami, Saori; Wakita, Ryo; Kohase, Hikaru; Saito, Norio; Yoshioka, Tomohiko; Ikoma, Toshiyuki; Tanaka, Junzo; Umino, Masahiro; Fukayama, Haruhisa

    2012-03-13

    Calcium alginate gel has some unique properties, such as the capability to keep the drugs, bioadhesiveness, safety, and low cost. The purpose of this study is to determine whether calcium alginate gel can be used as a matrix of electrodes for iontophoresis (IOP). We measured the concentration of lidocaine transported from calcium alginate gels with various concentrations of alginic acid using an in vitro experimental cell with square-wave alternating current (AC) application. Temperature and pH changes were also determined during AC-IOP. The results revealed that lidocaine was released from calcium alginate gels at concentrations nearly 1.71-fold larger at 5 V, 60 min after AC application than in the case of passive diffusion. Lidocaine transport depended on the alginic acid concentration in the gels. Although there were slight increases in temperature and pH, chemical and thermal burns were not severe enough to be a concern. In conclusion, the calcium alginate gel can be used as a possible matrix for IOP electrodes.

  16. Comparison of antimicrobial activities and compressive strength of alginate impression materials following disinfection procedure.

    Science.gov (United States)

    Alwahab, Zahraa

    2012-07-01

    This study investigated the effectiveness of disinfecting solution when incorporated into alginate powder instead of water against some microorganisms and on compressive strength of alginate. For measuring antimicrobial activity of alginate, 60 alginate specimens were prepared and divided into two groups: One with water incorporated in the mix (control) and the other with 0.2% chlorhexidine digluconate incorporated in the mix instead of water. The tested microorganisms were: gram +ve cocci, gram -ve bacilli and yeast (each group 10 samples). For measuring compressive strength, 20 specimens of alginate were divided into two groups: One with water incorporated in the mix (control) and the other with chlorhexidine incorporated in the mix. The statistical analysis of antimicrobial efficacy of alginate was performed with Mann-Whitney U-test, which revealed very high significant difference when comparing among groups (p 0.05). The incorporation of disinfecting agents into impression materials could serve an important role in dental laboratory infection control and it had no adverse effect on compressive strength of the hydrocolloid alginate. The risk of transmitting pathogenic microorganisms to dental laboratories via impression has been considered a topic of importance for a number of years.

  17. Effect of formulation of alginate beads on their mechanical behavior and stiffness

    Institute of Scientific and Technical Information of China (English)

    Eng-Seng Chan; Tek-Kaun Lim; Wan-Ping Voo; Ravindra Pogaku; Beng Ti Tey; Zhibing Zhang

    2011-01-01

    The aim of this work was to determine the effect of formulation of alginate beads on their mechanical behavior and stiffness when compressed at high speed. The alginate beads were formulated using different types and concentrations of alginate and gelling cations and were produced using an extrusiondripping method. Single wet beads were compressed at a speed of 40 mm/min, and their elastic limits were investigated, and the corresponding force versus displacement data were obtained. The Young's moduli of the beads were determined from the force versus displacement data using the Hertz's contact mechanics theory. The alginate beads were found to exhibit plastic behavior when they were compressed beyond 50% with the exception of copper-alginate beads for which yield occured at lower deformation.Alginate beads made of higher guluronic acid contents and gelling cations of higher chemical affinity were found to have greater stiffness. Increasing the concentration of alginate and gelling ions also generated a similar effect. At such a compression speed, the values of Young's modulus of the beads were found to be in the range between 250 and 900 kPa depending on the bead formulation.

  18. High surface area mesoporous activated carbon-alginate beads for efficient removal of methylene blue.

    Science.gov (United States)

    Nasrullah, Asma; Bhat, A H; Naeem, Abdul; Isa, Mohamed Hasnain; Danish, Mohammed

    2018-02-01

    High surface area mesoporous activated carbon-alginate (AC-alginate) beads were successfully synthesized by entrapping activated carbon powder derived from Mangosteen fruit peel into calcium-alginate beads for methylene blue (MB) removal from aqueous solution. The structure and surface characteristics of AC-alginate beads were analyzed using Fourier transform infra-red (FTIR) spectroscopy, scanning electron microscopy (SEM) and surface area analysis (S BET ), while thermal properties were tested using thermogravimetric analysis (TGA). The effect of AC-alginate dose, pH of solution, contact time, initial concentration of MB solution and temperature on MB removal was elucidated. The results showed that the maximum adsorption capacity of 230mg/g was achieved for 100mg/L of MB solution at pH 9.5 and temperature 25°C. Furthermore, the adsorption of MB on AC-alginate beads followed well pseudo-second order equation and equilibrium adsorption data were better fitted by the Freundlich isotherm model. The findings reveal the feasibility of AC-alginate beads composite to be used as a potential and low cost adsorbent for removal of cationic dyes. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Effect of Sodium Alginate Addition to Resveratrol on Acute Gouty Arthritis

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    Peng Wang

    2015-04-01

    Full Text Available Objective: Resveratrol has been shown to exert anti-inflammatory and antioxidant effects, while sodium alginate is a common pharmaceutic adjuvant with antioxidative and immunomodulatory properties. We performed an animal study to investigate the effect of sodium alginate addition to resveratrol on acute gouty arthritis. Methods: Twenty-four SPF Wistar mice were randomized to four groups receiving the combination of sodium alginate and resveratrol, resveratrol alone, colchicine, and placebo, respectively. Acute gouty arthritis was induced by injection of 0.05 ml monosodium urate (MSU solution (25g/mL into ankle joint cavity. IL-1β, CCR5, and CXCL10 levels in both serum and synovial fluid were measured using ELISA. NLRP3 expression in the synovial tissues was measured using western plot. Results: The combination of sodium alginate and resveratrol significantly reduced synovial levels of IL-1β, CCR5, and CXCL10 when compared with colchicines, and all P values were less than 0.0001. The combination of sodium alginate and resveratrol was also superior to resveratrol in terms of both serum levels and synovial levels of IL-1β, CCR5, and CXCL10. In addition, resveratrol, with or without sodium alginate, could reduce NLRP3 expression obviously in the synovial tissues. Conclusion: The combination of sodium alginate and resveratrol has better effect over colchicines in treating MSU-induced acute gouty arthritis.

  20. Highly Concentrated Alginate-Gellan Gum Composites for 3D Plotting of Complex Tissue Engineering Scaffolds

    Directory of Open Access Journals (Sweden)

    Ashwini Rahul Akkineni

    2016-04-01

    Full Text Available In tissue engineering, additive manufacturing (AM technologies have brought considerable progress as they allow the fabrication of three-dimensional (3D structures with defined architecture. 3D plotting is a versatile, extrusion-based AM technology suitable for processing a wide range of biomaterials including hydrogels. In this study, composites of highly concentrated alginate and gellan gum were prepared in order to combine the excellent printing properties of alginate with the favorable gelling characteristics of gellan gum. Mixtures of 16.7 wt % alginate and 2 or 3 wt % gellan gum were found applicable for 3D plotting. Characterization of the resulting composite scaffolds revealed an increased stiffness in the wet state (15%–20% higher Young’s modulus and significantly lower volume swelling in cell culture medium compared to pure alginate scaffolds (~10% vs. ~23%. Cytocompatibility experiments with human mesenchymal stem cells (hMSC revealed that cell attachment was improved—the seeding efficiency was ~2.5–3.5 times higher on the composites than on pure alginate. Additionally, the composites were shown to support hMSC proliferation and early osteogenic differentiation. In conclusion, print fidelity of highly concentrated alginate-gellan gum composites was comparable to those of pure alginate; after plotting and crosslinking, the scaffolds possessed improved qualities regarding shape fidelity, mechanical strength, and initial cell attachment making them attractive for tissue engineering applications.