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Sample records for pseudomonas fluorescens bacteria

  1. The effect of phylogenetically different bacteria on the fitness of Pseudomonas fluorescens in sand microcosms.

    Directory of Open Access Journals (Sweden)

    Olaf Tyc

    Full Text Available In most environments many microorganisms live in close vicinity and can interact in various ways. Recent studies suggest that bacteria are able to sense and respond to the presence of neighbouring bacteria in the environment and alter their response accordingly. This ability might be an important strategy in complex habitats such as soils, with great implications for shaping the microbial community structure. Here, we used a sand microcosm approach to investigate how Pseudomonas fluorescens Pf0-1 responds to the presence of monocultures or mixtures of two phylogenetically different bacteria, a Gram-negative (Pedobacter sp. V48 and a Gram-positive (Bacillus sp. V102 under two nutrient conditions. Results revealed that under both nutrient poor and nutrient rich conditions confrontation with the Gram-positive Bacillus sp. V102 strain led to significant lower cell numbers of Pseudomonas fluorescens Pf0-1, whereas confrontation with the Gram-negative Pedobacter sp. V48 strain did not affect the growth of Pseudomonas fluorescens Pf0-1. However, when Pseudomonas fluorescens Pf0-1 was confronted with the mixture of both strains, no significant effect on the growth of Pseudomonas fluorescens Pf0-1 was observed. Quantitative real-time PCR data showed up-regulation of genes involved in the production of a broad-spectrum antibiotic in Pseudomonas fluorescens Pf0-1 when confronted with Pedobacter sp. V48, but not in the presence of Bacillus sp. V102. The results provide evidence that the performance of bacteria in soil depends strongly on the identity of neighbouring bacteria and that inter-specific interactions are an important factor in determining microbial community structure.

  2. The impact of cellulose nanocrystals on the aggregation and initial adhesion of Pseudomonas fluorescens bacteria.

    Science.gov (United States)

    Sun, Xiaohui; Lu, Qingye; Boluk, Yaman; Liu, Yang

    2014-11-28

    Deposition on silica surfaces of two Pseudomonas fluorescens strains (CHA0 and CHA19-WS) having different extracellular polymeric substance (EPS) producing capacities was studied in the absence and presence of cellulose nanocrystals (CNCs). Batch (batch soaking) and continuous flow (quartz crystal microbalance with dissipation) methods were used to evaluate the impact of CNCs on bacterial initial adhesion. This study demonstrated that bacterial initial adhesion to solid surfaces can be significantly hindered by CNCs using both methods. In the presence of CNCs, it was observed that bacteria with more EPS aggregated more significantly compared to bacteria with less EPS, and that bacterial deposition under this condition decreased to a greater extent. The classic DLVO theory failed to predict bacterial adhesion behavior in this study. A detailed discussion is provided regarding potential antibacterial adhesion mechanisms of CNCs.

  3. 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...

  4. 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.

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

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    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. 

  6. Plant growth promotion by Pseudomonas fluorescens

    NARCIS (Netherlands)

    Cheng, X.

    2016-01-01

    Pseudomonas fluorescens is a Gram-negative rod shaped bacterium that has a versatile metabolism and is widely spread in soil and water. P. fluorescens strain SBW25 (Pf.SBW25) is a well-known model strain to study bacterial evolution, plant colonization and biocontrol of plant diseases. It produces

  7. The effect of phylogenetically different bacteria on the fitness of Pseudomonas fluorescens in sand microcosms

    NARCIS (Netherlands)

    Tyc, Olaf; Wolf, Alexandra; Garbeva, Paolina

    2015-01-01

    n most environments many microorganisms live in close vicinity and can interact in various ways. Recent studies suggest that bacteria are able to sense and respond to the presence of neighbouring bacteria in the environment and alter their response accordingly. This ability might be an important

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

    African Journals Online (AJOL)

    This involved hydrolysis of starch extracted from freshly harvested cassava tubers using enzyme-enzyme method of hydrolysis, followed by aerobic fermentation of Pseudomonas fluorescens on a mixture of the hydrolysate and nutrient media in a fermentor in batch cultures. The reducing sugar hydrolysate served as the ...

  9. 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.

  10. 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.

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

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    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.

  12. 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...

  13. Bacteriocins and the assembly of natural Pseudomonas fluorescens populations.

    Science.gov (United States)

    Bruce, J B; West, S A; Griffin, A S

    2017-02-01

    When competing for space and resources, bacteria produce toxins known as bacteriocins to gain an advantage over competitors. Recent studies in the laboratory have confirmed theoretical predictions that bacteriocin production can determine coexistence, by eradicating sensitive competitors or driving the emergence of resistant genotypes. However, there is currently limited evidence that bacteriocin-mediated competition influences the coexistence and distribution of genotypes in natural environments, and what factors drive interactions towards inhibition remain unclear. Using natural soil populations of Pseudomonas fluorescens, we assessed the ability of the isolates to inhibit one another with respect to spatial proximity in the field, genetic similarity and niche overlap. The majority of isolates were found to produce bacteriocins; however, widespread resistance between coexisting isolates meant relatively few interactions resulted in inhibition. When inhibition did occur, it occurred more frequently between ecologically similar isolates. However, in contrast with results from other natural populations, we found no relationship between the frequency of inhibition and the genetic similarity of competitors. Our results suggest that bacteriocin production plays an important role in mediating competition over resources in natural settings and, by selecting for isolates resistant to local bacteriocin production, can influence the assembly of natural populations of P. fluorescens. © 2016 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2016 European Society For Evolutionary Biology.

  14. 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.

  15. 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 ...

  16. Characterization and identification of Pseudomonas fluorescens ...

    African Journals Online (AJOL)

    user

    2011-01-24

    Jan 24, 2011 ... fluorescens can be used for bioremediation of textile effluents containing CV dye in sequential cycles. Key words: Crystal violet, ... extensively in textile dyeing and dye-stuff manufacturing industries, in human and ... nuclear magnetic resonance spectra of the test samples were run at Brukar AC 200 M Hz in ...

  17. Lack of AHL-based quorum sensing in Pseudomonas fluorescens isolated from milk

    Science.gov (United States)

    Martins, Maurilio L.; Pinto, Uelinton M.; Riedel, Kathrin; Vanetti, Maria C.D.; Mantovani, Hilário C.; de Araújo, Elza F.

    2014-01-01

    Numerous bacteria coordinate gene expression in response to small signalling molecules in many cases known as acylhomoserine lactones (AHLs), which accumulate as a function of cell density in a process known as quorum sensing. This work aimed to determine if phenotypes that are important to define microbial activity in foods such as biofilm formation, swarming motility and proteolytic activity of two Pseudomonas fluorescens strains, isolated from refrigerated raw milk, are influenced by AHL molecules. The tested P. fluorescens strains did not produce AHL molecules in none of the evaluated media. We found that biofilm formation was dependent on the culture media, but it was not influenced by AHLs. Our results indicate that biofilm formation, swarming motility and proteolytic activity of the tested P. fluorescens strains are not regulated by acyl-homoserine lactones. It is likely that AHL-dependent quorum sensing system is absent from these strains. PMID:25477941

  18. Impact of a Recombinant Biocontrol Bacterium, Pseudomonas fluorescens pc78, on Microbial Community in Tomato Rhizosphere.

    Science.gov (United States)

    Kong, Hyun Gi; Kim, Nam Hee; Lee, Seung Yeup; Lee, Seon-Woo

    2016-04-01

    Pseudomonas fluorescens pc78 is an effective biocontrol agent for soil-borne fungal diseases. We previously constructed a P43-gfp tagged biocontrol bacteria P. fluorescens pc78-48 to investigate bacterial traits in natural ecosystem and the environmental risk of genetically modified biocontrol bacteria in tomato rhizosphere. Fluctuation of culturable bacteria profile, microbial community structure, and potential horizontal gene transfer was investigated over time after the bacteria treatment to the tomato rhizosphere. Tagged gene transfer to other organisms such as tomato plants and bacteria cultured on various media was examined by polymerase chain reaction, using gene specific primers. Transfer of chromosomally integrated P43-gfp from pc78 to other organisms was not apparent. Population and colony types of culturable bacteria were not significantly affected by the introduction of P. fluorescens pc78 or pc78-48 into tomato rhizosphere. Additionally, terminal restriction fragment length polymorphism profiles were investigated to estimate the influence on the microbial community structure in tomato rhizosphere between non-treated and pc78-48-treated samples. Interestingly, rhizosphere soil treated with strain pc78-48 exhibited a significantly different bacterial community structure compared to that of non-treated rhizosphere soil. Our results suggest that biocontrol bacteria treatment influences microbial community in tomato rhizosphere, while the chromosomally modified biocontrol bacteria may not pose any specific environmental risk in terms of gene transfer.

  19. EXAFS Study of Uranyl Complexation at Pseudomonas fluorescens Cell Surfaces

    Science.gov (United States)

    Bencheikh, R.; Bargar, J. R.; Tebo, B. M.

    2002-12-01

    Little is known about the roles of microbial biomass as a sink and source for uranium in contaminated aquifers, nor of the impact of bacterial biochemistry on uranium speciation in the subsurface. A significant role is implied by the high affinities of both Gram positive and Gram negative cells for binding uranyl (UO2{ 2+}). In the present study, Extended X-ray Absorption Fine Structure (EXAFS) spectroscopy was used to identify membrane functional groups involved in uranyl binding to the Gram negative bacterium Pseudomonas fluorescens from pH 3 to pH 8. Throughout this pH-range, EXAFS spectra can be described primarily in terms of coordination of carboxylic groups to uranyl. U-C distances characteristic of 4-, 5- and 8- membered rings were observed, as well as the possibility of phosphato groups. Both shell-by-shell fits and principle component analyses indicate that the functional groups involved in binding of uranyl to the cell surface do not vary systematically across the pH range investigated. This result contrasts with EXAFS results of uranyl sorbed to Gram positive bacteria, and suggests an important role for long-chain carboxylate-terminated membrane functional groups in binding uranyl.

  20. 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.

  1. The evolution of a pleiotropic fitness tradeoff in Pseudomonas fluorescens

    OpenAIRE

    MacLean, R. Craig; Bell, Graham; Rainey, Paul B.

    2004-01-01

    The evolution of ecological specialization is expected to carry a cost, due to either antagonistic pleiotropy or mutation accumulation. In general, it has been difficult to distinguish between these two possibilities. Here, we demonstrate that the experimental evolution of niche-specialist genotypes of the bacterium Pseudomonas fluorescens that colonize the air–broth interface of spatially structured microcosms is accompanied by pleiotropic fitness costs in terms of reduced carbon catabolism....

  2. Inhibition of quorum sensing-controlled virulence factors and biofilm formation in Pseudomonas fluorescens by cinnamaldehyde.

    Science.gov (United States)

    Li, Tingting; Wang, Dangfeng; Liu, Nan; Ma, Yan; Ding, Ting; Mei, Yongchao; Li, Jianrong

    2018-02-01

    Pseudomonas fluorescens, an important food spoiling bacteria, uses quorum sensing to control biofilm formation and motility. To date, only a few compounds targeting the LuxR-based quorum sensing system of P. fluorescens have been identified. In the present study, the quorum sensing inhibitory effect of cinnamaldehyde at sublethal concentrations was investigated in terms of inhibition of the extracellular protease, biofilm formation, and swimming and swarming motility. The total volatile basic nitrogen value was also measured to evaluate the effect of cinnamaldehyde on quality preservation of turbot fillets stored at 4 ± 1 °C for 15 days. The results showed that cinnamaldehyde significantly inhibited quorum sensing-dependent factors in P. fluorescens and extended the storage life of turbot. Unexpectedly, cinnamaldehyde did not interfere with production of AHLs (N-acylhomoserine lactones) by P. fluorescens, as shown by measurement of AHL production using GC-MS. Molecular docking analysis revealed that cinnamaldehyde can interact with the LuxR-type protein of P. fluorescens, which could constitute the molecular basis of the quorum sensing inhibition observed. These findings strongly suggest that cinnamaldehyde is a quorum sensing inhibitor with great potential for the preservation of aquatic products to guarantee food safety. Copyright © 2018. Published by Elsevier B.V.

  3. 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...

  4. Pseudomonas fluorescens subsp. cellulosa: an alternative model for bacterial cellulase.

    Science.gov (United States)

    Hazlewood, G P; Laurie, J I; Ferreira, L M; Gilbert, H J

    1992-03-01

    Pseudomonas fluorescens subsp. cellulosa, a Gram-negative soil bacterium, can utilize crystalline cellulose or xylan as main sources of carbon and energy. Synthesis of endoglucanases and xylanases is induced by Avicel, filter paper, carboxymethylcellulose or xylan and is repressed by cellobiose, glucose or xylose. These enzymes are secreted into the culture supernatant fluid and do not form aggregates or associate with the cell surface. Cells of Ps. fluorescens subsp. cellulosa do not adhere to cellulose. In cultures containing Avicel or filter paper, a significant proportion of the secreted cellulase and xylanase activities becomes tightly bound to the insoluble cellulose. Western blotting has revealed that endoglucanase B, xylanase A and a cellodextrinase encoded by genes previously isolated from Ps. fluorescens subsp. cellulosa and expressed in Escherichia coli, are synthesized by the pseudomonad under a variety of conditions. These enzymes appear to be post-translationally modified, probably through glycosylation. Overall, it appears that the cellulase/hemicellulase system of Ps. fluorescens subsp. cellulosa differs from the model established for celluloytic anaerobes such as Clostridium thermocellum.

  5. MICROBIAL BIOFILMS PRODUCED BY PSEUDOMONAS FLUORESCENS ON SOLID SURFACES

    Directory of Open Access Journals (Sweden)

    Dagmar Kozelová

    2011-04-01

    Full Text Available Normal 0 21 false false false MicrosoftInternetExplorer4 Normal 0 21 false false false MicrosoftInternetExplorer4 A biofilm is a complex aggregation of microorganisms growing on a solid substrate. Biofilms are characterized by structural heterogeneity, genetic diversity, complex community interactions, and an extracellular matrix of polymeric substances. 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 four test surfaces of stainless steel, glass and plastic materials - PE (polyethylene and EPDM (ethylene propylene diene monomer. Over the next 72 hours and 72 hours were observed numbers of adhesion bacterial cells of P. fluorescens on solid surfaces of tested materials. The highest values adhesion cells reached P. fluorescens cells after 72 hours of cultivation on plastic surfaces, where  was increased in adhesion bacterial cells for EPDM in the values of 105 CFU/cm2 and for PE up to 106 CFU/cm2. The subsequent repeated 72-hour cultivation P. fluorescens was an increase (growth in the number of adhesion bacterial cells to all tested surfaces.doi:10.5219/18  

  6. 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.

  7. 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.

  8. 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...... 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...... can drive adaptive evolution and suggest that this class of mutation may be underappreciated as a cause of adaptation and evolutionary dynamics....

  9. Uji Lapangan Formula Cair Pseudomonas fluorescens P60 terhadap Layu Fusarium pada Tanaman Tomat

    Directory of Open Access Journals (Sweden)

    Loekas Soesanto

    2011-12-01

    Full Text Available A research aimed at knowing 1 the effect of Pseudomonas fluorescens P60 in liquid formula on Fusarium wilt of tomato, 2 the effect of P. fluorescens P60 in the formula on tomato growth and yield, and 3 P. fluorescens P60 mechanisms on tomata was carried out at tomato field of Selomoyo Village, Kaliangkrik Subdistrict, Magelang Regency at altitude of 826 m above sea level. Randomized block design was used with seven treatments and four replicates. The treatments were control, with P. fluorescens P60 soaked for 15 min and without fungicide, pathogen without P. fluorescens P60 with fungicide (PBG1, pathogen with P. fluorescens P60 without fungicide, pathogen with pouring P. fluorescens P60 1, 3, and 5 times. Result indicated that application of formulated P. fluorescens P60 for 5 times decreased the disease intensity as high as 26.77%, and late population of the pathogen but increased P. fluorescens P60 as high as 4.54×1010 cfu ml-1. P. fluorescens P60 affected growth and yield of tomato. P. Fluorescens P60 induced tomato resistance by increasing qualitatively its phenolic compound content (saponin, tannin, glycoside.   Penelitian dengan tujuan untuk mengetahui: 1 pengaruh Pseudomonas fluorescens P60 dalam formula cair terhadap penyakit layu fusarium pada tanaman tomat, 2 pengaruh P. fluorescens P60 dalam formula cair terhadap pertumbuhan dan produksi tanaman tomat, dan 3 mekanisme P. fluorescens P60 pada tanaman tomat dilakukan di lahan Desa Selomoyo, Kecamatan Kaliangkrik, Kabupaten Magelang dengan ketinggian 826 m di atas permukaan laut. Penelitian menggunakan Rancangan Acak Kelompok (RAK, dengan 7 perlakuan dan jumlah ulangan 4 kali, dan setiap unit terdiri atas 8 tanaman. Perlakuan tersebut meliputi kontrol; dengan P. fluorescens P60 rendam 15 menit dan tanpa fungisida; dengan patogen; tanpa P. fluorescens P60; dengan fungisida (PBG1; patogen, tanpa P. fluorescens P60, tanpa fungisida; patogen, dengan penyiraman P. fluorescens P60 1 kali

  10. 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

  11. Analysis of Bacillus thuringiensis Population Dynamics and Its Interaction With Pseudomonas fluorescens in Soil

    Science.gov (United States)

    Rojas-Ruiz, Norma Elena; Sansinenea-Royano, Estibaliz; Cedillo-Ramirez, Maria Lilia; Marsch-Moreno, Rodolfo; Sanchez-Alonso, Patricia; Vazquez-Cruz, Candelario

    2015-01-01

    Background: Bacillus thuringiensis is the most successful biological control agent, however, studies so far have shown that B. thuringiensis is very sensitive to environmental factors such as soil moisture and pH. Ultraviolet light from the sun had been considered as the main limiting factor for its persistence in soil and it has recently been shown that the antagonism exerted by other native soil organisms, such as Pseudomonas fluorescens, is a determining factor in the persistence of this bacterium under in vitro culture conditions. Objectives: The aim of the present investigation was to analyze the population dynamics of B. thuringiensis and its interaction with P. fluorescens using microbiological and molecular methods in soil, under different conditions, and to determinate the effect of nutrients and moisture on its interaction. Materials and Methods: The monitoring was performed by microbiological methods, such as viable count of bacteria, and molecular methods such as Polymerase Chain Reaction (PCR) and hybridization, using the direct extraction of DNA from populations of inoculated soil. Results: The analysis of the interaction between B. thuringiensis and P. fluorescens in soil indicated that the disappearance of B. thuringiensis IPS82 is not dependent on the moisture but the composition of nutrients that may be affecting the secretion of toxic compounds in the environment of P. fluorescens. The results showed that the recovered cells were mostly spores and not vegetative cells in all proved treatments. The molecular methods were effective for monitoring bacterial population inoculated in soil. Conclusions: Bacillus thuringiensis is very sensitive to the interaction of P. fluorescens, however is capable to survive in soil due to its capacity of sporulate. Some of the cells in the form of spores germinated and folded slightly and remained in a constant cycle of sporulation and germination. This confirms that B. thuringiensis IPS82 can germinate, grow and

  12. Analysis of Bacillus thuringiensis Population Dynamics and Its Interaction With Pseudomonas fluorescens in Soil.

    Science.gov (United States)

    Rojas-Ruiz, Norma Elena; Sansinenea-Royano, Estibaliz; Cedillo-Ramirez, Maria Lilia; Marsch-Moreno, Rodolfo; Sanchez-Alonso, Patricia; Vazquez-Cruz, Candelario

    2015-09-01

    Bacillus thuringiensis is the most successful biological control agent, however, studies so far have shown that B. thuringiensis is very sensitive to environmental factors such as soil moisture and pH. Ultraviolet light from the sun had been considered as the main limiting factor for its persistence in soil and it has recently been shown that the antagonism exerted by other native soil organisms, such as Pseudomonas fluorescens, is a determining factor in the persistence of this bacterium under in vitro culture conditions. The aim of the present investigation was to analyze the population dynamics of B. thuringiensis and its interaction with P. fluorescens using microbiological and molecular methods in soil, under different conditions, and to determinate the effect of nutrients and moisture on its interaction. The monitoring was performed by microbiological methods, such as viable count of bacteria, and molecular methods such as Polymerase Chain Reaction (PCR) and hybridization, using the direct extraction of DNA from populations of inoculated soil. The analysis of the interaction between B. thuringiensis and P. fluorescens in soil indicated that the disappearance of B. thuringiensis IPS82 is not dependent on the moisture but the composition of nutrients that may be affecting the secretion of toxic compounds in the environment of P. fluorescens. The results showed that the recovered cells were mostly spores and not vegetative cells in all proved treatments. The molecular methods were effective for monitoring bacterial population inoculated in soil. Bacillus thuringiensis is very sensitive to the interaction of P. fluorescens, however is capable to survive in soil due to its capacity of sporulate. Some of the cells in the form of spores germinated and folded slightly and remained in a constant cycle of sporulation and germination. This confirms that B. thuringiensis IPS82 can germinate, grow and sporulate in soil.

  13. Longitudinal nosocomial outbreak of Pseudomonas fluorescens bloodstream infection of 2 years' duration in a coronary care unit.

    Science.gov (United States)

    Oba, Yuichiro; Nakajima, Takahiro; Ogida, Chiyo; Kawanami, Miyuki; Fujiwara, Mamoru; Matsumura, Ikuko

    2017-08-01

    Outbreaks of bloodstream infections (BSI) of nonfermenting bacteria are a critical issue and often associated with hospital environments. We experienced a long-lasting outbreak of Pseudomonas fluorescens BSI limited to a coronary care unit (CCU). We conducted a retrospective epidemiologic investigation and a case-control study for Pseudomonas fluorescens BSI from April 2011-July 2014. Environmental sample culture was conducted to detect the specific environmental source of transmission. Hospital-wide microbiology data from the term identified 13 case patients with P fluorescens BSI and 32 control patients with BSI due to organisms other than P fluorescens in the CCU. The case-control study revealed that the case group had significantly higher odds of exposure to only cardiac output (CO) measurement with thermodilution method (odds ratio, 22.0; 95% confidence interval, 2.4-202.3). The organism was identified only from an ice bath used for CO measurement. The susceptibility patterns were identical among all strains derived from the cases and the environment. The nosocomial outbreak of P fluorescens BSI in our CCU over 2 years was associated with a contaminated ice bath used for CO measurement within the unit. Detection and elimination of the specific source was essential to stop the outbreak. Copyright © 2017 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Elsevier Inc. All rights reserved.

  14. Isolation and characterization of a T7-like lytic phage for Pseudomonas fluorescens

    OpenAIRE

    Neubauer Peter; Sillankorva Sanna; Azeredo Joana

    2008-01-01

    Background: Despite the proven relevance of Pseudomonas fluorescens as a spoilage microorganism in milk, fresh meats and refrigerated food products and the recognized potential of bacteriophages as sanitation agents, so far no phages specific for P. fluorescens isolates from dairy industry have been closely characterized in view of their lytic efficiency. Here we describe the isolation and characterization of a lytic phage capable to infect a variety of P. fluorescens strains i...

  15. 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

  16. 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.

  17. Lethality and developmental delay in Drosophila melanogaster larvae after ingestion of selected Pseudomonas fluorescens strains.

    Directory of Open Access Journals (Sweden)

    Marika H Olcott

    2010-09-01

    Full Text Available The fruit fly, Drosophila melanogaster, is a well-established model organism for probing the molecular and cellular basis of physiological and immune system responses of adults or late stage larvae to bacterial challenge. However, very little is known about the consequences of bacterial infections that occur in earlier stages of development. We have infected mid-second instar larvae with strains of Pseudomonas fluorescens to determine how infection alters the ability of larvae to survive and complete development.We mimicked natural routes of infection using a non-invasive feeding procedure to study the toxicity of the three sequenced P. fluorescens strains (Pf0-1, SBW25, and Pf-5 to Drosophila melanogaster. Larvae fed with the three strains of P. fluorescens showed distinct differences in developmental trajectory and survival. Treatment with SBW25 caused a subset of insects to die concomitant with a systemic melanization reaction at larval, pupal or adult stages. Larvae fed with Pf-5 died in a dose-dependent manner with adult survivors showing eye and wing morphological defects. In addition, larvae in the Pf-5 treatment groups showed a dose-dependent delay in the onset of metamorphosis relative to control-, Pf0-1-, and SBW25-treated larvae. A functional gacA gene is required for the toxic properties of wild-type Pf-5 bacteria.These experiments are the first to demonstrate that ingestion of P. fluorescens bacteria by D. melanogaster larvae causes both lethal and non-lethal phenotypes, including delay in the onset of metamorphosis and morphological defects in surviving adult flies, which can be decoupled.

  18. Pseudomonas fluorescens pirates both ferrioxamine and ferricoelichelin siderophores from Streptomyces ambofaciens.

    Science.gov (United States)

    Galet, Justine; Deveau, Aurélie; Hôtel, Laurence; Frey-Klett, Pascale; Leblond, Pierre; Aigle, Bertrand

    2015-05-01

    Iron is essential in many biological processes. However, its bioavailability is reduced in aerobic environments, such as soil. To overcome this limitation, microorganisms have developed different strategies, such as iron chelation by siderophores. Some bacteria have even gained the ability to detect and utilize xenosiderophores, i.e., siderophores produced by other organisms. We illustrate an example of such an interaction between two soil bacteria, Pseudomonas fluorescens strain BBc6R8 and Streptomyces ambofaciens ATCC 23877, which produce the siderophores pyoverdine and enantiopyochelin and the siderophores desferrioxamines B and E and coelichelin, respectively. During pairwise cultures on iron-limiting agar medium, no induction of siderophore synthesis by P. fluorescens BBc6R8 was observed in the presence of S. ambofaciens ATCC 23877. Cocultures with a Streptomyces mutant strain that produced either coelichelin or desferrioxamines, as well as culture in a medium supplemented with desferrioxamine B, resulted in the absence of pyoverdine production; however, culture with a double mutant deficient in desferrioxamines and coelichelin production did not. This strongly suggests that P. fluorescens BBbc6R8 utilizes the ferrioxamines and ferricoelichelin produced by S. ambofaciens as xenosiderophores and therefore no longer activates the production of its own siderophores. A screening of a library of P. fluorescens BBc6R8 mutants highlighted the involvement of the TonB-dependent receptor FoxA in this process: the expression of foxA and genes involved in the regulation of its biosynthesis was induced in the presence of S. ambofaciens. In a competitive environment, such as soil, siderophore piracy could well be one of the driving forces that determine the outcome of microbial competition. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  19. 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-...

  20. 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.

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

    Science.gov (United States)

    Timm, Collin M; Campbell, Alisha G; Utturkar, Sagar M; Jun, Se-Ran; Parales, Rebecca E; Tan, Watumesa A; Robeson, Michael S; Lu, Tse-Yuan S; Jawdy, Sara; Brown, Steven D; Ussery, David W; Schadt, Christopher W; Tuskan, Gerald A; Doktycz, Mitchel J; Weston, David J; Pelletier, Dale A

    2015-01-01

    The bacterial microbiota of plants is diverse, with 1000s of operational taxonomic units (OTUs) associated with any individual plant. In this work, we used phenotypic analysis, comparative genomics, and metabolic models to investigate the differences between 19 sequenced Pseudomonas fluorescens strains. These isolates represent a single OTU and were collected from the rhizosphere and endosphere of Populus deltoides. 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 toward 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 toward 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.

  2. 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.

  3. Bioreporter pseudomonas fluorescens HK44 immobilized in a silica matrix

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    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.

  4. Nonribosomal peptides, key biocontrol components for Pseudomonas fluorescens In5, isolated from a Greenlandic suppressive soil.

    Science.gov (United States)

    Michelsen, Charlotte F; Watrous, Jeramie; Glaring, Mikkel A; Kersten, Roland; Koyama, Nobuhiro; Dorrestein, Pieter C; Stougaard, Peter

    2015-03-17

    Potatoes are cultivated in southwest Greenland without the use of pesticides and with limited crop rotation. Despite the fact that plant-pathogenic fungi are present, no severe-disease outbreaks have yet been observed. In this report, we document that a potato soil at Inneruulalik in southern Greenland is suppressive against Rhizoctonia solani Ag3 and uncover the suppressive antifungal mechanism of a highly potent biocontrol bacterium, Pseudomonas fluorescens In5, isolated from the suppressive potato soil. A combination of molecular genetics, genomics, and matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) imaging mass spectrometry (IMS) revealed an antifungal genomic island in P. fluorescens In5 encoding two nonribosomal peptides, nunamycin and nunapeptin, which are key components for the biocontrol activity by strain In5 in vitro and in soil microcosm experiments. Furthermore, complex microbial behaviors were highlighted. Whereas nunamycin was demonstrated to inhibit the mycelial growth of R. solani Ag3, but not that of Pythium aphanidermatum, nunapeptin instead inhibited P. aphanidermatum but not R. solani Ag3. Moreover, the synthesis of nunamycin by P. fluorescens In5 was inhibited in the presence of P. aphanidermatum. Further characterization of the two peptides revealed nunamycin to be a monochlorinated 9-amino-acid cyclic lipopeptide with similarity to members of the syringomycin group, whereas nunapeptin was a 22-amino-acid cyclic lipopeptide with similarity to corpeptin and syringopeptin. Crop rotation and systematic pest management are used to only a limited extent in Greenlandic potato farming. Nonetheless, although plant-pathogenic fungi are present in the soil, the farmers do not experience major plant disease outbreaks. Here, we show that a Greenlandic potato soil is suppressive against Rhizoctonia solani, and we unravel the key biocontrol components for Pseudomonas fluorescens In5, one of the potent biocontrol bacteria

  5. The cellodextrinase from Pseudomonas fluorescens subsp. cellulosa consists of multiple functional domains

    National Research Council Canada - National Science Library

    Ferreira, L M; Hazlewood, G P; Barker, P J; Gilbert, H J

    1991-01-01

    A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA was constructed in pUC18 and Escherichia coli recombinants expressing 4-methylumbelliferyl beta-D-cellobioside-hydrolysing activity (MUCase) were isolated...

  6. 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

  7. 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

  8. 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.

  9. Saccharomyces cerevisiae genome-wide mutant screen for sensitivity to 2,4-diacetylphloroglucinol, a biocontrol antibiotic produced by Pseudomonas fluorescens

    Science.gov (United States)

    2,4-diacetylphloroglucinol (2,4-DAPG) is an antibiotic produced by Pseudomonas fluorescens that plays a key role in the ability of the bacterium to suppress phytopathogenic fungi. 2,4-DAPG has broad antibiotic activity, affecting organisms ranging from bacteria to higher plants. The biosynthesis and...

  10. Pseudomonas fluorescens SBW25 produces furanomycin, a non-proteinogenic amino acid with selective antimicrobial properties

    Science.gov (United States)

    2013-01-01

    Background Pseudomonas fluorescens SBW25 has been extensively studied because of its plant growth promoting properties and potential as a biocontrol agent. The genome of SBW25 has been sequenced, and among sequenced strains of pseudomonads, SBW25 appears to be most closely related to P. fluorescens WH6. In the authors’ laboratories, WH6 was previously shown to produce and secrete 4-formylaminooxyvinylglycine (FVG), a non-proteinogenic amino acid with selective herbicidal and antimicrobial activity. Although SBW25 does not have the genetic capacity to produce FVG, we were interested in determining whether this pseudomonad might produce some other type of non-proteinogenic amino acid. Results P. fluorescens SBW25 was found to produce and secrete a ninhydrin-reactive compound with selective antimicrobial properties. This compound was purified from SBW25 culture filtrate and identified as the non-proteinogenic amino acid L-furanomycin [2S,2′R,5′S)-2-amino-2-(5′methyl-2′,5′-dihydrofuran-2′-yl)acetic acid]. Conclusions The identification of furanomycin as a secondary metabolite of SBW25 is the first report of the production of furanomycin by a pseudomonad. This compound was known previously only as a natural product produced by a strain of Streptomyces. This report adds furanomycin to the small list of non-proteinogenic amino acids that have been identified as secondary products of pseudomonads. This study also extends the list of bacteria that are inhibited by furanomycin to include several plant pathogenic bacteria. PMID:23688329

  11. Effect of a Pseudomonas fluorescens tailocin against phytopathogenic Xanthomonas observed by atomic force microscopy.

    Science.gov (United States)

    Fernandez, Maricruz; Godino, Agustina; Príncipe, Analía; Morales, Gustavo M; Fischer, Sonia

    2017-08-20

    Phage tail-like bacteriocins, called tailocins, represent a class of protein complexes produced by a multitude of bacteria. Pseudomonas fluorescens SF4c, a strain isolated from wheat rhizosphere, produces a bacteriocin similar to phage tail-like pyocins of Pseudomonas aeruginosa. This tailocin has antimicrobial activity against several phytopathogenic strains of the genus Xanthomonas and Pseudomonas. In this work, the effect of the SF4c tailocin on the phytopathogenic strain X. axonopodis pv vesicatoria Xcv Bv5-4a was analyzed through Atomic Force Microscopy (AFM). We demonstrated that tailocins adhere and cause damage to the cell envelope of strain Xcv Bv5-4a. This results in a rapid leakage of intracellular materials, with the subsequent decrease of cell volume. Finally, lysis of sensitive bacteria occurs. This study provides, to our knowledge, the first evidence about the effect of a tailocin analyzed by AFM. Further studies are in progress to evaluate the use of SF4c tailocin in the biocontrol of bacterial spot on tomato. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Diluent sensitivity in thermally stressed cells of pseudomonas fluorescens.

    Science.gov (United States)

    Gray, R J; Ordal, Z J; Witter, L D

    1977-01-01

    Thermally injured cells of Pseudomonas fluorescens were unable to produce colonies on Trypticase soy agar (TSA) after dilution with 0.1% peptone. Nutritional exigency could not be used as the criterion for this injury, since varying the composition of the plating medium had little effect on the number of colonies that developed. The injured cells had no requirement for compounds known to leak out during the heat treatment in order to recover. The cells did not exhibit injury if dilution preceded heat treatment on the plating medium, demonstrating that the heat treatment sensitized the cells to the trauma of dilution. Substitution of 0.1% peptone with growth medium as the diluent largely offset the previously observed drop in TSA count. Little difference in survival was observed when monosodium glutamate or the balance of the defined medium was used as the diluent. The diluent effect was ionic rather than osmotic. The presence of cations was important in maintaining the integrity of the injured cell, and divalent cations enhanced this protective effect. The role of these cations at the level of the cell envelope is discussed. PMID:406839

  13. Global control in Pseudomonas fluorescens mediating antibiotic synthesis and suppression of black root rot of tobacco.

    OpenAIRE

    Laville, J; Voisard, C; C. Keel; Maurhofer, M.; Défago, G; Haas, D

    1992-01-01

    Pseudomonas fluorescens CHA0 colonizes plant roots, produces several secondary metabolites in stationary growth phase, and suppresses a number of plant diseases, including Thielaviopsis basicola-induced black root rot of tobacco. We discovered that mutations in a P. fluorescens gene named gacA (for global antibiotic and cyanide control) pleiotropically block the production of the secondary metabolites 2,4-diacetylphloroglucinol (Phl), HCN, and pyoluteorin. The gacA mutants of strain CHA0 have...

  14. Bioremediation of crude oil contaminated soil by bioaugmentation of Pseudomonas fluorescens NS1.

    Science.gov (United States)

    Barathi, S; Vasudevan, N

    2003-09-01

    A feasibility study was conducted to evaluate the efficiency of Pseudomonas fluorescens NS1 bioaugmented to stimulate in situ bioremediation of crude oil-contaminated soil with different amendments in treatment units. Pure culture of P. fluorescens NS1 was isolated from a petroleum-contaminated soil. The rate of degradation of petroleum hydrocarbons by the indigenous soil microflora and in the presence of P. fluorescens NS1 was assessed with the addition of nutrients and bulking agents for a period of about 35 days. The study showed that addition of wheat bran as bulking agent rapidly enhanced bioremediation of crude oil-contaminated soil compared to amendments in other treatment units.

  15. Biocontrol of collar rot disease of betelvine (Piper betle L.) caused by Sclerotium rolfsii by using rhizosphere-competent Pseudomonas fluorescens NBRI-N6 and P. fluorescens NBRI-N.

    Science.gov (United States)

    Singh, Anand; Mehta, Sangeeta; Singh, Harikesh Bahadur; Nautiyal, Chandra Shekhar

    2003-08-01

    Collar rot disease of betelvine (Piper betle L.) caused by Sclerotium rolfsii is difficult to control by conventional means by use of chemicals; therefore, use of biocontrol agents is desirable. In the present study, 186 bacterial strains of different morphological types were screened for their biocontrol activity against S. rolfsii under in vitro conditions. Two strains, Pseudomonas fluorescens NBRI-N6 and P. fluorescens NBRI-N, were selected for further studies because of their ability to inhibit the mycelial growth of the pathogen significantly. Spontaneous rifampicin-resistant (Rif) derivatives of P. fluorescens NBRI-N6 and P. fluorescens NBRI-N showing growth rate and membrane protein composition comparable to the wild type were selected to facilitate their monitoring in the rhizosphere. Field trials demonstrated that strain P. fluorescens NBRI-N6 was better than P. fluorescens NBRI-N in increasing the yield of betelvine significantly, whereas a consortium of the two strains controlled the disease more than either of the strains. The screening method should prove useful in identifying rhizosphere bacteria with the greatest potential for controlling diseases caused by phytopathogenic fungi.

  16. Transposon Tn5 mutagenesis of pseudomonas fluorescens to isolate mutants deficient in antibacterial activity.

    Science.gov (United States)

    Rajendran, N; Jahn, D; Jayaraman, K; Marahiel, M A

    1994-01-15

    Pseudomonas fluorescens was subjected to insertion mutagenesis studies using the transposon Tn5-GM to generate mutants deficient in antibacterial activity minus mutants. The transposon located on the temperature-sensitive plasmid pCHR84 was conjugally transferred into the non-pathogenic pseudomonad using the triparental mating procedure. Random integration of Tn5-GM into the chromosome of P. fluorescens was achieved by heat treatment of the transformed cells at 42 degrees C. Approximately 2% of transconjugants revealed an auxotrophic phenotype indicating efficient integration of the employed transposon into the chromosome of P. fluorescens. One transposon insertion mutant was obtained showing an antibacterial activity minus phenotype. This mutant (MM-7) was found to be defective in the production of an unidentified antibacterial compound against B. subtilis. These results introduce Tn5 transposon mutagenesis as a new useful tool for the molecular analysis of P. fluorescens.

  17. Chromium phytoextraction from tannery effluent-contaminated soil by Crotalaria juncea infested with Pseudomonas fluorescens.

    Science.gov (United States)

    Agarwal, Anamika; Singh, Harminder Pal; Rai, J P N

    2014-01-01

    The aim of present study was to remediate chromium (Cr)-contaminated soil by Crotalaria juncea in the presence of Pseudomonas fluorescens. Inoculation of P. fluorescens in pot soil grown with C. juncea significantly increased (~2-fold) the water-soluble (Ws) and exchangeable (Ex) Cr contents in contaminated soil under greenhouse condition. It also enhanced the chlorophyll content by 92 % and plant biomass by 99 % as compared to the uninoculated C. juncea plant. The analysis showed that root and shoot uptake of Cr in C. juncea inoculated by P. fluorescens was 3.08- and 2.82-fold, respectively. This research showed that the association of C. juncea and P. fluorescens could be a promising technology for increasing the soil Cr bioavailability and plant growth for successful phytoextraction of Cr from the contaminated soil.

  18. On the role of metabolic activity on the transport und deposition of Pseudomonas fluorescens in saturated porous media.

    Science.gov (United States)

    Jansen, Sandra; Vereecken, Harry; Klumpp, Erwin

    2010-02-01

    A study was conducted to understand the role of cell concentration and metabolic state in the transport and deposition behaviour of Pseudomonas fluorescens with and without substrate addition. Column experiments using the short-pulse technique (pulse was equivalent to 0.028 pore volume) were performed in quartz sand operating under saturated conditions. For comparison, experiments with microspheres and inactive (killed) bacteria were also conducted. The effluent concentrations, the retained particle concentrations and the cell shape were determined by fluorescent microscopy. For the transport of metabolically-active P. fluorescens without substrate addition a bimodal breakthrough curve was observed, which could be explained by the different breakthrough behaviour of the rod-shaped and coccoidal cells of P. fluorescens. The 70:30 rod/coccoid ratio in the influent drastically changed during the transport and it was about 20:80 in the effluent and in the quartz sand packing. It was assumed that the active rod-shaped cells were subjected to shrinkage into coccoidal cells. The change from active rod-shaped cells to coccoidal cells could be explained by oxygen deficiency which occurs in column experiments under saturated conditions. Also the substrate addition led to two consecutive breakthrough peaks and to more bacteria being retained in the column. In general, the presence of substrate made the assumed stress effects more pronounced. In comparison to microspheres and inactive (killed) bacteria, the transport of metabolically-active bacteria with and without substrate addition is affected by differences in physiological state between rod-shaped and the formed stress-resistant coccoidal cells of P. fluorescens. Copyright 2010 Elsevier Ltd. All rights reserved.

  19. Colonisation of Pinus halepensis roots by Pseudomonas fluorescens and interaction with the ectomycorrhizal fungus Suillus granulatus.

    Science.gov (United States)

    Rincón, Ana; Ruiz-Díez, Beatriz; García-Fraile, Sonia; García, José Antonio Lucas; Fernández-Pascual, Mercedes; Pueyo, José J; de Felipe, María R

    2005-02-01

    Colonisation of Pinus halepensis roots by GFP-tagged Pseudomonas fluorescens Aur6 was monitored by epifluorescence microscopy and dilution plating. Aur6-GFP was able to colonise and proliferate on P. halepensis roots. Co-inoculation with the ectomycorrhizal fungus Suillus granulatus did not affect the bacterial colonisation pattern whereas it had an effect on bacterial density. Bacterial counts increased during the first 20 days of seedling growth, irrespective of seedlings being mycorrhizal or not. After 40 days, bacterial density significantly decreased and bacteria concentrated on the upper two-thirds of the pine root. The presence of S. granulatus significantly stimulated survival of bacteria in the root elongation zone where fungal colonisation was higher. The number of mycorrhizas formed by S. granulatus was not affected by co-inoculation with Aur6-GFP. Neither Aur6-GFP nor S. granulatus stimulated P. halepensis development when inoculated alone, but a synergistic effect was observed on seedling growth when bacteria and fungus were co-inoculated.

  20. 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

  1. Genome sequence of the mycorrhizal helper bacterium Pseudomonas fluorescens BBc6R8

    Energy Technology Data Exchange (ETDEWEB)

    Deveau, Aurelie [French National Insitute for Agricultural Research (INRA); Grob, Harald [University of Bonn, Germany; Morin, Emmanuelle [INRA, Nancy, France; Karpinets, Tatiana V [ORNL; Utturkar, Sagar M [ORNL; Mehnaz, Samina [University of the Punjab, Pakistan; Kurz, Sven [University of Bonn, Germany; Martin, Francis [INRA, Nancy, France; Frey-Klett, Pascale [INRA, Nancy, France; Labbe, Jessy L [ORNL

    2014-01-01

    We report the draft genome sequence of the mycorrhiza helper bacterium Pseudomonas fluorescens strain BBc6R8 . Several traits which could be involved in the mycorrhiza helper ability of the bacterial strain such as multiple secretion systems, auxin metabolism and phosphate mobilization were evidenced in the genome.

  2. 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...

  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. 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

  5. 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,

  6. Application of Pseudomonas fluorescens to Blackberry under Field Conditions Improves Fruit Quality by Modifying Flavonoid Metabolism.

    Directory of Open Access Journals (Sweden)

    Daniel Garcia-Seco

    Full Text Available Application of a plant growth promoting rhizobacterium (PGPR, Pseudomonas fluorescens N21.4, to roots of blackberries (Rubus sp. is part of an optimised cultivation practice to improve yields and quality of fruit throughout the year in this important fruit crop. Blackberries are especially rich in flavonoids and therefore offer potential benefits for human health in prevention or amelioration of chronic diseases. However, the phenylpropanoid pathway and its regulation during ripening have not been studied in detail, in this species. PGPR may trigger flavonoid biosynthesis as part of an induced systemic response (ISR given the important role of this pathway in plant defence, to cause increased levels of flavonoids in the fruit. We have identified structural genes encoding enzymes of the phenylpropanoid and flavonoid biosynthetic pathways catalysing the conversion of phenylalanine to the final products including flavonols, anthocyanins and catechins from blackberry, and regulatory genes likely involved in controlling the activity of pathway branches. We have also measured the major flavonols, anthocyanins and catechins at three stages during ripening. Our results demonstrate the coordinated expression of flavonoid biosynthetic genes with the accumulation of anthocyanins, catechins, and flavonols in developing fruits of blackberry. Elicitation of blackberry plants by treatment of roots with P.fluorescens N21.4, caused increased expression of some flavonoid biosynthetic genes and an accompanying increase in the concentration of selected flavonoids in fruits. Our data demonstrate the physiological mechanisms involved in the improvement of fruit quality by PGPR under field conditions, and highlight some of the genetic targets of elicitation by beneficial bacteria.

  7. Extracellular enzyme production and cheating in Pseudomonas fluorescens depend on diffusion rates.

    Science.gov (United States)

    Allison, Steven D; Lu, Lucy; Kent, Alyssa G; Martiny, Adam C

    2014-01-01

    Bacteria produce extracellular enzymes to obtain resources from complex chemical substrates, but this strategy is vulnerable to cheating by cells that take up reaction products without paying the cost of enzyme production. We hypothesized that cheating would suppress enzyme production in co-cultures of cheater and producer bacteria, particularly under well-mixed conditions. To test this hypothesis, we monitored protease expression and frequencies of Pseudomonas fluorescens producer and cheater genotypes over time in mixed liquid cultures and on agar plates. In mixed culture inoculated with equal frequencies of cheaters and producers, enzyme concentration declined to zero after 20 days, consistent with our hypothesis. We observed a similar decline in cultures inoculated with producers only, suggesting that cheater mutants arose de novo and swept the population. DNA sequencing showed that genetic changes most likely occurred outside the protease operon. In one experimental replicate, the population regained the ability to produce protease, likely due to further genetic changes or population dynamics. Under spatially structured conditions on agar plates, cheaters did not sweep the population. Instead, we observed a significant increase in the variation of enzyme activity levels expressed by clones isolated from the population. Together these results suggest that restricted diffusion favors a diversity of enzyme production strategies. In contrast, well-mixed conditions favor population sweeps by cheater strains, consistent with theoretical predictions. Cheater and producer strategies likely coexist in natural environments with the frequency of cheating increasing with diffusion rate.

  8. Legionella pneumophila persists within biofilms formed by Klebsiella pneumoniae, Flavobacterium sp., and Pseudomonas fluorescens under dynamic flow conditions.

    Directory of Open Access Journals (Sweden)

    Catherine R Stewart

    Full Text Available Legionella pneumophila, the agent of Legionnaires' disease pneumonia, is transmitted to humans following the inhalation of contaminated water droplets. In aquatic systems, L. pneumophila survives much of time within multi-organismal biofilms. Therefore, we examined the ability of L. pneumophila (clinical isolate 130 b to persist within biofilms formed by various types of aquatic bacteria, using a bioreactor with flow, steel surfaces, and low-nutrient conditions. L. pneumophila was able to intercalate into and persist within a biofilm formed by Klebsiella pneumoniae, Flavobacterium sp. or Pseudomonas fluorescens. The levels of L. pneumophila within these biofilms were as much as 4 × 10(4 CFU per cm(2 of steel coupon and lasted for at least 12 days. These data document that K. pneumoniae, Flavobacterium sp., and P. fluorescens can promote the presence of L. pneumophila in dynamic biofilms. In contrast to these results, L. pneumophila 130 b did not persist within a biofilm formed by Pseudomonas aeruginosa, confirming that some bacteria are permissive for Legionella colonization whereas others are antagonistic. In addition to colonizing certain mono-species biofilms, L. pneumophila 130 b persisted within a two-species biofilm formed by K. pneumoniae and Flavobacterium sp. Interestingly, the legionellae were also able to colonize a two-species biofilm formed by K. pneumoniae and P. aeruginosa, demonstrating that a species that is permissive for L. pneumophila can override the inhibitory effect(s of a non-permissive species.

  9. 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

  10. Di-Adenosine Tetraphosphate (Ap4A) Metabolism Impacts Biofilm Formation by Pseudomonas fluorescens via Modulation of c-di-GMP-Dependent Pathways▿

    OpenAIRE

    Monds, Russell D.; Newell, Peter D.; Wagner, Jeffrey C.; Schwartzman, Julia A.; Lu, Wenyun; Rabinowitz, Joshua D.; O'Toole, George A.

    2010-01-01

    Dinucleoside tetraphosphates are common constituents of the cell and are thought to play diverse biological roles in organisms ranging from bacteria to humans. In this study we characterized two independent mechanisms by which di-adenosine tetraphosphate (Ap4A) metabolism impacts biofilm formation by Pseudomonas fluorescens. Null mutations in apaH, the gene encoding nucleoside tetraphosphate hydrolase, resulted in a marked increase in the cellular level of Ap4A. Concomitant with this increase...

  11. The structure-function relationship of WspR, a Pseudomonas fluorescens response regulator with a GGDEF output domain

    National Research Council Canada - National Science Library

    Malone, J. G; Williams, R; Christen, M; Jenal, U; Spiers, A. J; Rainey, P. B

    2007-01-01

    ...{at}auckland.ac.nz The GGDEF response regulator WspR couples the chemosensory Wsp pathway to the overproduction of acetylated cellulose and cell attachment in the Pseudomonas fluorescens SBW25 wrinkly spreader (WS) genotype...

  12. HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS

    Directory of Open Access Journals (Sweden)

    ANTONIUS SUWANTO

    2003-01-01

    Full Text Available A transcriptional fusion for an Aeromonas caviae chitinase gene was constructed under the control of a constitutive promoter of the kanaraycin resistance gene (PKmR. The construct was inserted into a medium copy number broad host range plasmid vector to yield recombinant plasmid pAM340, which harbored transcriptional fusion PKmR- chi. Another transcriptional fusion, Ptac-chi, in a recombinant plasmid pAM630, was conducted as comparison. Triparental mating of E. coli carrying the recombinant plasmids with Pseudomotws fluorescens 5100, a phyllosphere bacterium, was performed. Pseudomonas fluorescens 5100 exconjugants were examined for constitutive expression of chitinase employing a spectrophotometric assay; they showed stronger chitin degradation activity than Escherichia coli transformants. Using a fungal antagonism plate assay, this chitinolytic P. fluorescens, however, could not inhibit selected phytopathogenic fungi.

  13. Systematic investigations on the biodegradation and viscosity reduction of long chain hydrocarbons using Pseudomonas aeruginosa and Pseudomonas fluorescens.

    Science.gov (United States)

    Sakthipriya, N; Doble, Mukesh; Sangwai, Jitendra S

    2016-03-01

    The use of microorganisms has been researched extensively for possible applications related to hydrocarbon degradation in the petroleum industry. However, attempts to improve the effect of microorganisms on the viscosity of hydrocarbons, which find potential use in the development of robust models for biodegradation, have been rarely documented. This study investigates the degradation of long chain hydrocarbons, such as hexadecane and eicosane using Pseudomonas fluorescens PMMD3 (P. fluorescens) and Pseudomonas aeruginosa CPCL (P. aeruginosa). P. aeruginosa used here is isolated from petroleum contaminated sediments and the P. fluorescens is from the coastal area, and both have hydrocarbon degrading genes. The degradation of hydrocarbons is studied using carbon profiling and reduction in viscosity pre- and post-degradation of hydrocarbons. The carbon profiling has been obtained using gas chromatography-mass spectroscopy (GC-MS), and Fourier transform infrared spectrometer (FTIR) results. GC-MS results have indicated an improved biodegradation of hydrocarbons by 77-93% in one day. The yield coefficients of biomass (YX/S) for P. aeruginosa and P. fluorescens using hexadecane as a carbon source are 1.35 and 0.81 g g(-1), and the corresponding values with eicosane are 0.84 and 0.88 g g(-1). The viscosity of hexadecane is reduced by the order of 53 and 47%, while that of eicosane was reduced by 53 and 65%, using P. aeruginosa and P. fluorescens, respectively. This study also presents information on the activity of enzymes responsible for the hydrocarbon degradation. Pseudomonas species have shown their use in potential applications for bioremediation, oil-spill treatment, and flow assurance. We believe that this study will also provide stringent tests for possible model development for the bioremediation of long chain paraffins suitable for oilfield applications.

  14. Pseudomine, an isoxazolidone with siderophoric activity from Pseudomonas fluorescens AH2 isolated from Lake Victorian Nile Perch

    DEFF Research Database (Denmark)

    Anthoni, U.; Christophersen, C.; Nielsen, P.H.

    1995-01-01

    A siderophore, pseudomonine, and sodium salicylate were isolated from the culture broth of iron-deficient cultures of Pseudomonas fluorescens AH2 isolated from the surface of spoiled Nile Perch from Lake Victoria......A siderophore, pseudomonine, and sodium salicylate were isolated from the culture broth of iron-deficient cultures of Pseudomonas fluorescens AH2 isolated from the surface of spoiled Nile Perch from Lake Victoria...

  15. 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.

  16. Aplikasi Formula Cair Pseudomonas fluorescens P60 untuk Menekan Penyakit Virus Cabai Merah

    Directory of Open Access Journals (Sweden)

    Loekas Soesanto

    2014-08-01

    Full Text Available Viral diseases of chilli pepper are difficult to control, therefore the use of Pseudomonas fluorescens P60 should be evaluated. The aims of this research were to determine the influence of liquid formula of P. fluorescens P60 on virod disease and on growth and yield of chili pepper. Randomized block design (RBD experiment was composed of 7 treatments and 4 replicates, i.e., control, insecticide applicaton, P. fluorescens P60 application by seedling drenching and spraying for 1, 2, 3, 5, and 7 times. The result showed that 5 times application of P. fluorescens P60 by drenching and spraying was able to suppress viral disease and reduce disease intensity by to 73.37%, increasing density level of P. fluorescens P60 to 9.50 x 1011 and increase phenolic compounds (saponin, tannin and glycoside. The same treatment could increase plant height 23.7%, root lenght 6.44%, plant dry weight 66.68%, root dry weight 23.59%, and yield weight 53.16%.

  17. Cheating and resistance to cheating in natural populations of the bacterium Pseudomonas fluorescens.

    Science.gov (United States)

    Bruce, John B; Cooper, Guy A; Chabas, Hélène; West, Stuart A; Griffin, Ashleigh S

    2017-10-01

    Bacteria perform cooperative behaviors that are exploitable by noncooperative cheats, and cheats frequently arise and coexist with cooperators in laboratory microcosms. However, evidence of competitive dynamics between cooperators and cheats in nature remains limited. Using the production of pyoverdine, an iron-scavenging molecule, and natural soil populations of Pseudomonas fluorescens, we found that (1) nonproducers are present in the population; (2) they co-occur (cheats in soil: utilizing the pyoverdine of others while producing little or none themselves. However, we found considerable variation in the extent to which nonproducers can exploit producers, as some isolates appear to produce exclusive forms of pyoverdine or kill nonproducers with toxins. We examined the consequences of this variation using theoretical modeling. We found variance in exploitability leads to some cheats gaining increased fitness benefits and others decreased benefits. However, the absolute gain in fitness from high exploitation is lower than the drop in fitness from low exploitation, decreasing the mean fitness of cheats and subsequently lowering the proportion of cheats maintained in the population. Our results suggest that although cooperator-cheat dynamics can occur in soil, a range of mechanisms can prevent nonproducers from exploiting producers. © 2017 The Author(s). Evolution published by Wiley Periodicals, Inc. on behalf of The Society for the Study of Evolution.

  18. Milk-deteriorating exoenzymes from Pseudomonas fluorescens 041 isolated from refrigerated raw milk

    Science.gov (United States)

    Martins, Maurilio L.; Pinto, Uelinton M.; Riedel, Katharina; Vanetti, Maria C.D.

    2015-01-01

    The practice of refrigerating raw milk at the farm has provided a selective advantage for psychrotrophic bacteria that produce heat-stable proteases and lipases causing severe quality problems to the dairy industry. In this work, a protease (AprX) and a lipase (LipM) produced by Pseudomonas fluorescens 041, a highly proteolytic and lipolytic strain isolated from raw milk obtained from a Brazilian farm, have been purified and characterized. Both enzymes were purified as recombinant proteins from Escherichia coli . The AprX metalloprotease exhibited activity in a broad temperature range, including refrigeration, with a maximum activity at 37 °C. It was active in a pH range of 4.0 to 9.0. This protease had maximum activity with the substrates casein and gelatin in the presence of Ca +2 . The LipM lipase had a maximum activity at 25 °C and a broad pH optimum ranging from 7.0 to 10. It exhibited the highest activity, in the presence of Ca +2 , on substrates with long-chain fatty acid residues. These results confirm the spoilage potential of strain 041 in milk due to, at least in part, these two enzymes. The work highlights the importance of studies of this kind with strains isolated in Brazil, which has a recent history on the implementation of the cold chain at the dairy farm. PMID:26221110

  19. 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)

  20. Effect of exogenous ectoines on some antifungal activity of Pseudomonas fluorescens UTPF5 under salt conditions

    Directory of Open Access Journals (Sweden)

    Arghavan Kamaly

    2016-06-01

    Full Text Available Introduction: Plant probiotic bacteria like fluorescent pseudomonads are worldly used against soil-borne pathogens through mechanisms such as production of bacterial metabolites and enzymes. These bacteria can also help plants to tolerate environmental stresses. Ectoine is a compatible solution which plays an important role in environmental osmotic stresses. Materials and methods: Tolerance of 24 bacterial strains to salt and heat was tested and 6 tolerant strains were chosen. Then dual culture of Fusarium solani and bacteria grown in presence of ectoine and hydroxy ectoine with 3 NaCl concentrations (0, 150 and 300 mM was done and Pseudomonas fluorescens UTPF5 was selected. Finally the effect of ectoine, hydroxyectoine and NaCl on bacterial population, lipase, protease, siderophore and hydrogen-cyanide production and biofilm formation was investigated. Results: In 300 mM NaCl, the bacterium grown in presence of ectoines, increased the inhibition percentage 5- times more than control. NaCl had a positive effect on bacterial population in water and ectoine, hydrogen-cyanide production in all treatment and biofilm formation in ectoine and hydroxyectoine and negative effect on bacterial population in hydroxyectoine, protease and siderophore production in all treatments and biofilm formation in water. On the other hand, ectoine increased lipase and hydrogen-cyanide production and biofilm formation in 300 mM NaCl and siderophore production in 150 mM. Hydroxyectoine had similar effects with little differences. Discussion and conclusion: Ectoine and hydroxyectoine moderate the biocontrol reduction in presence of NaCl through positive effect on lipase and hydrogen-cyanide production and biofilm formation.

  1. 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.

  2. 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 and

  3. The cellodextrinase from Pseudomonas fluorescens subsp. cellulosa consists of multiple functional domains.

    OpenAIRE

    Ferreira, L M; Hazlewood, G P; Barker, P J; Gilbert, H J

    1991-01-01

    A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA was constructed in pUC18 and Escherichia coli recombinants expressing 4-methylumbelliferyl beta-D-cellobioside-hydrolysing activity (MUCase) were isolated. Enzyme produced by MUCase-positive clones did not hydrolyse either cellobiose or cellotriose but converted cellotetraose into cellobiose and cleaved cellopentaose and cellohexaose, producing a mixture of cellobiose and cellotriose. There was no activity against CM-cellulose,...

  4. Cyanase-mediated utilization of cyanate in Pseudomonas fluorescens NCIB 11764.

    OpenAIRE

    Kunz, D. A.; Nagappan, O

    1989-01-01

    Pseudomonas fluorescens NCIB 11764 was capable of utilizing cyanate (OCN-) as a sole nitrogen source for growth. Crude cell extracts from cells grown on cyanate, but not on ammonium sulfate, were induced for an enzyme catalyzing cyanate conversion to ammonia. Enzymatic activity was shown to be bicarbonate dependent and specific for cyanate as a substrate, suggesting that cyanate utilization in this organism is facilitated by an enzyme resembling cyanase (cyanate amidohydrolase; EC 3.5.5.3), a...

  5. Adhesion of Pseudomonas fluorescens biofilms to glass, stainless steel and cellulose

    OpenAIRE

    Wan Dagang, W.R.Z.; Bowen, J.; O'Keeffe, J.; Robbins, P.T.; Zhang, Z.

    2016-01-01

    Objectives: \\ud The adhesion of colloidal probes of stainless steel, glass and cellulose to Pseudomonas fluorescens biofilms was examined using atomic force microscopy (AFM) to allow comparisons between surfaces to which biofilms might adhere.\\ud \\ud Results: \\ud Biofilm was grown on a stainless steel substrate and covered most of the surface after 96 h. AFM approach and retraction curves were obtained when the biofilm was immersed in a tryptone/soy medium. On approach, all the colloidal prob...

  6. Effect of Pseudomonas fluorescens RB4 and Bacillus subtilis 189 on the phytoremediation potential of Catharanthus roseus (L.) in Cu and Pb-contaminated soils.

    Science.gov (United States)

    Khan, Waheed Ullah; Ahmad, Sajid Rashid; Yasin, Nasim Ahmad; Ali, Aamir; Ahmad, Aqeel

    2017-06-03

    The remediation of heavy metal-contaminated soils has become a critical issue due to toxic effects of these metals on living organisms. The current research was conducted to study the effect of Pseudomonas fluorescens RB4 and Bacillus subtilis 189 on the growth and phytoremediation potential of Catharanthus roseus in Cu- and Pb-contaminated soils. The bacterial strains exhibited significantly higher level of water-extractable Pb and Cu in Pb, Cu, and Cu+Pb-contaminated. The P. fluorescens RB4 inoculated plants, produced 102%, 48%, and 45% higher fresh weight (FW) in soils contaminated with Cu, Pb, and both elements, respectively, as compared to un-inoculated control plants. Similarly, B. subtilis 189 inoculated plants produced 108%, 43%, and 114% more FW in the presence of Cu, Pb, and both elements. The plants co-cultivated with both bacteria exhibited 121%, 102%, and 177% higher FW, in Cu, Pb, and both elements contaminated soils, as compared to respective un-inoculated control. Co-cultivation of P. fluorescens RB4, B. subtilis 189, and P. fluorescens RB4 + B. subtilis 189 resulted in higher accumulation of Cu and Pb in shoots of the C. roseus grown in contaminated soils as compared to un-inoculated control. Bacterial treatments also improved the translocation and metal bioconcentration factors. The growth and phytoextraction capability of C. roseus was improved by inoculation of P. fluorescens RB4 and B. subtilis 189.

  7. Biocontrol strain Pseudomonas fluorescens WCS365 inhibits germination of Fusarium oxysporum spores in tomato root exudate as well as subsequent formation of new spores.

    Science.gov (United States)

    Kamilova, Faina; Lamers, Gerda; Lugtenberg, Ben

    2008-09-01

    Fusarium oxysporum f.sp.radicis-licopersici (Forl) is a soilborne pathogenic fungus which can cause tomato foot and root rot (TFRR). Tomato root exudate is a good source of nutrients for both Forl and the TFRR-suppressing biocontrol bacterium Pseudomonas fluorescens strain WCS365. Incubation of Forl microconidia in tomato root exudate stimulates their germination. This phenomenon is observed, to a lesser extent, upon incubation in plant nutrient solution supplemented with citrate or glucose, the major organic acid and sugar components, respectively, of tomato root exudate. Here we show that induction of germination of microconidia is significantly reduced in the presence of P. fluorescens WCS365 in all tested media. Scanning electron microscopy revealed that P. fluorescens WCS365 colonizes developing hyphae. Efficient colonization correlates with low nutrient availability. Eventually, new microconidia are formed. The presence of P. fluorescens WCS365 reduces the number of newly formed microconidia. This reduction does not depend on physical contact between bacteria and hyphae. We discuss that the ability of P. fluorescens WCS365 to slow down the processes of microconidia germination and development of new microconidia of the phytopathogen, and therefore the ability to reduce fungal dissemination, is likely to contribute to the biocontrol efficacy of this strain.

  8. A genetically engineered Pseudomonas fluorescens strain possesses the dual activity against phytopathogenic fungi and insects.

    Science.gov (United States)

    Lu, Wenwei; Zhang, Weiqiong; Bai, Yan; Fu, Yingying; Chen, Jun; Geng, Xiaolu; Wang, Yujing; Xiao, Ming

    2010-02-01

    A Pseudomonas fluorescens strain was isolated and showed antagonistic activity against phytopathogenic fungi and found to possess a gene responsible for production of antibiotic 2, 4-diacetylphloroglucinol. For the extension of biocontrol range, a gene for an Androctonus australis Hector insect toxin 1 (AaHIT1), one of the most toxic known insect-selective peptides, was designed and synthesized according to the preferred codon usage of Pseudomonas fluorescens, cloned and transformed into the strain by pSUP106 vector, a broad-host-range plasmid. Bioassays indicated that the engineered strain was able to produce AaHIT1 with insecticidal activity, in the same time retained the activity against plant pathogen. The experiments for nonplanted soil and rhizosphere colonization showed that, similar to the population of the wild-type strain, that of the engineered strain remained relatively constant in the first 10 d, and the subsequent 50 d, suggesting that AaHIT expression in the bacterial cell does not substantially impair its long-term colonization. It is first reported that a Pseudomonas fluorescens strain expressing an active scorpion neurotoxin has dual activity against phytopathogenic fungi and insects, attractive for agronomic applications.

  9. Initial characterization of a bolA homologue from Pseudomonas fluorescens indicates different roles for BolA-like proteins in em>P. fluorescens and Escherichia coli

    DEFF Research Database (Denmark)

    Koch, Birgit; Nybroe, Ole

    2006-01-01

    The RpoS-regulated bolA gene in Escherichia coli is important for the decrease in cell size during stationary phase or sudden carbon starvation. A Pseudomonas fluorescens strain mutated in a gene with homology to bolA reduced its cell size upon carbon starvation, and RpoS had little effect on bol...

  10. Isolation and characterization of a T7-like lytic phage for Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    Neubauer Peter

    2008-10-01

    Full Text Available Abstract Background Despite the proven relevance of Pseudomonas fluorescens as a spoilage microorganism in milk, fresh meats and refrigerated food products and the recognized potential of bacteriophages as sanitation agents, so far no phages specific for P. fluorescens isolates from dairy industry have been closely characterized in view of their lytic efficiency. Here we describe the isolation and characterization of a lytic phage capable to infect a variety of P. fluorescens strains isolated from Portuguese and United States dairy industries. Results Several phages were isolated which showed a different host spectrum and efficiency of lysis. One of the phages, phage ϕIBB-PF7A, was studied in detail due to its efficient lysis of a wide spectrum of P. fluorescens strains and ribotypes. Phage ϕIBB-PF7A with a head diameter of about 63 nm and a tail size of about 13 × 8 nm belongs morphologically to the Podoviridae family and resembles a typical T7-like phage, as analyzed by transmission electron microscopy (TEM. The phage growth cycle with a detected latent period of 15 min, an eclipse period of 10 min, a burst size of 153 plaque forming units per infected cell, its genome size of approximately 42 kbp, and the size and N-terminal sequence of one of the protein bands, which gave similarity to the major capsid protein 10A, are consistent with this classification. Conclusion The isolated T7-like phage, phage ϕIBB-PF7A, is fast and efficient in lysing different P. fluorescens strains and may be a good candidate to be used as a sanitation agent to control the prevalence of spoilage causing P. fluorescens strains in dairy and food related environments.

  11. Effects of phosphorus fertilizer rate and Pseudomonas fluorescens strain on field pea (Pisum sativum subsp. arvense (L. Asch. growth and yield

    Directory of Open Access Journals (Sweden)

    Bahram SALEHI

    2015-11-01

    Full Text Available A field experiment was conducted at Rezvanshahr, Guilan province, Iran, to evaluate the effects of phosphorus fertilizer rate and Pseudomonas fluorescens strains on growth and yield of field pea (Pisum sativum L.. The experimental design was a randomized complete block in a factorial arrangement with three replicates. Factors were phosphorus fertilizer rates (0, 25, 50, 75, and 100 kg P2O5 ha-1 as triple superphosphate, and seed inoculation with P. florescens strains [control (non-inoculated, inoculated with strain R41, and strain R187. Analysis of variance showed that plant height, seed yield, pod number per m2, 100-seed weight, biological yield, harvest index, and leaf P concentration were significantly influenced by phosphorus fertilizer rate and P. florescens strain. At the same time, phosphorus fertilizer rate × P. fluorescens strain interaction was significant only for 100-seed weight. On the other hand, seed number per pod was significantly affected neither by phosphorus fertilizer rate nor by pseudomonas strains. Result showed that seed yield was significantly increased from 1099 ± 67 to 1898 ± 118 kg ha-1 as P2O5 application rate increased from 0 to 75 kg ha-1, and thereafter relatively remained constant. There was no significant difference in seed yield between plants raised from inoculated seeds with P. fluorescens, strain R187 (1664 ± 97 kg ha-1 and those raised from inoculated seeds with P. fluorescens, strain R41 (1669 ± 104 kg ha-1. At the same time, plants raised from inoculated seeds with P. fluorescens (both strains produced greater grain yield compared to those raised from uninoculated seeds (1370 ± 80 kg ha-1. Based on the results of this study, P2O5 application at the rate of 75 kg ha-1 and inoculation with pseudomonas bacteria are recommended for obtaining the greatest seed yield in field pea.

  12. [Pseudomonas genus bacteria on weeds].

    Science.gov (United States)

    Gvozdiak, R I; Iakovleva, L M; Pasichnik, L A; Shcherbina, T N; Ogorodnik, L E

    2005-01-01

    It has been shown in the work that the weeds (couch-grass and ryegrass) may be affected by bacterial diseases in natural conditions, Pseudomonas genus bacteria being their agents. The isolated bacteria are highly-aggressive in respect of the host-plant and a wide range of cultivated plants: wheat, rye, oats, barley, apple-tree and pear-tree. In contrast to highly aggressive bacteria isolated from the affected weeds, bacteria-epi phytes isolated from formally healthy plants (common amaranth, orache, flat-leaved spurge, field sow thistle, matricary, common coltsfoot, narrow-leaved vetch) and identified as P. syringae pv. coronafaciens, were characterized by weak aggression. A wide range of ecological niches of bacteria evidently promote their revival and distribution everywhere in nature.

  13. Genome-wide analysis of bacterial determinants of plant growth promotion and induced systemic resistance by Pseudomonas fluorescens

    NARCIS (Netherlands)

    Cheng, Xu; Etalo, Desalegn W.; van de Mortel, Judith E.; Dekkers, Ester; Nguyen, Linh; Medema, Marnix H; Raaijmakers, Jos M.

    2017-01-01

    Pseudomonas fluorescens strain SS101 (Pf.SS101) promotes growth of Arabidopsis thaliana, enhances greening and lateral root formation, and induces systemic resistance (ISR) against the bacterial pathogen Pseudomonas syringae pv. tomato (Pst). Here, targeted and untargeted approaches were adopted to

  14. 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.

  15. Investigation for zoonotic disease pathogens (Aeromonas hydrophila, Pseudomonas fluorescens, Streptococcus iniae) seen in carp farms in Duhok region of Northern Iraq by molecular methods

    Science.gov (United States)

    Mohammed, Kamiran Abdulrahman; Arabacı, Muhammed; Önalan, Şükrü

    2017-04-01

    The aim of this study was to determine the zoonotic bacteria in carp farms in Duhok region of the Northern Iraq. Carp is the main fish species cultured in the Duhok region. The most common zoonotic bacteria generally seen in carp farms are Aeromonas hydrophila, Pseudomonas fluorescens and Streptococcus iniae. Samples were collected from 20 carp farms in the Duhok Region of the Northern Iraq. Six carp samples were collected from each carp farm. Head kidney tissue samples and intestine tissue samples were collected from each carp sample. Than head kidney and intestine tissue samples were pooled. The total bacterial DNA extraction from the pooled each 20 head kidney tissue samples and pooled each 20 intestinal tissue samples. Primers for pathogens were originally designed from 16S Ribosomal gene region. Zoonotic bacteria were scanned in all tissue samples by absent / present analysis in the RT-PCR. After RT-PCR, Capillary gel electrophoresis bands were used for the confirmation of the size of amplicon which was planned during primer designing stage. As a result, one sample was positive in respect to Aeromonas hydrophila, from intestine and one carp farm was positive in respect to Pseudomonas fluorescens from intestine and two carp farms were positive in respect to Streptococcus iniae. Totally 17 of 20 carp farms were negative in respect to the zoonotic bacteria. In conclusion the zoonotic bacteria were very low (15 %) in carp farms from the Duhok Region in the Northern Iraq. Only in one Carp farms, both Aeromonas hydrophila and Pseudomonas fluorescens were positive. Also Streptococcus inia were positive in two carp farms.

  16. 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.)

  17. 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

  18. 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

    Proteins may be an important carbon and nitrogen source to bacteria in aquatic habitats, yet knowledge on the actual utilization of this substrate by proteolytic bacteria is scarce. In the present study, Pseudomonas fluorescens ON2 produced an alkaline proteinase (AprX) during growth and there wa......Proteins may be an important carbon and nitrogen source to bacteria in aquatic habitats, yet knowledge on the actual utilization of this substrate by proteolytic bacteria is scarce. In the present study, Pseudomonas fluorescens ON2 produced an alkaline proteinase (AprX) during growth...... the proteinase during growth unless a preferred carbon source like citrate is present. Lake water model systems were subsequently used to investigate the ability of proteolytic versus non-proteolytic ON2 strains to utilize protein for growth at moderate cell densities. Only cells forming surface-attached micro...

  19. Cyanase-mediated utilization of cyanate in Pseudomonas fluorescens NCIB 11764.

    Science.gov (United States)

    Kunz, D A; Nagappan, O

    1989-01-01

    Pseudomonas fluorescens NCIB 11764 was capable of utilizing cyanate (OCN-) as a sole nitrogen source for growth. Crude cell extracts from cells grown on cyanate, but not on ammonium sulfate, were induced for an enzyme catalyzing cyanate conversion to ammonia. Enzymatic activity was shown to be bicarbonate dependent and specific for cyanate as a substrate, suggesting that cyanate utilization in this organism is facilitated by an enzyme resembling cyanase (cyanate amidohydrolase; EC 3.5.5.3), as described previously in Escherichia coli and Flavobacterium sp.

  20. 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 (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).

  1. The combined effects of Pseudomonas fluorescens and Tuber melanosporum on the quality of Pinus halepensis seedlings.

    Science.gov (United States)

    Dominguez, J A; Martin, A; Anriquez, A; Albanesi, A

    2012-08-01

    The ecological, economic and social values of the ectomycorrhizal fungi of the black truffle found in the rural Mediterranean are well known. The inoculation of Pinus halepensis seedlings with mycorrhizal fungi and rhizobacteria can improve the morphology and physiology of the seedlings and benefit the regeneration of arid regions and the reintroduction of inocula of mycorrhizal fungi into these areas. Some rhizobacteria can improve the establishment and functioning of ectomycorrhizal symbiosis. In this study, seedlings of P. halepensis were inoculated with the mycorrhizal fungus Tuber melanosporum and the rhizobacteria Pseudomonas fluorescens CECT 844 under non-limiting greenhouse conditions. Five months after inoculation, we analysed the growth, water parameters (osmotic potential at saturation, osmotic potential at turgor loss and modulus of elasticity), concentrations of mycorrhizal colonies, nutrient concentration and nutrient contents (N, P, K, Ca, Mg and Fe) in roots and aerial parts of the seedlings. Subsequently, tests were performed to estimate the root growth potentials. None of the treatments changed the water parameters or growth potentials of the roots. The inoculations improved the growth and nutrient uptake of the seedlings, although the combination of P. fluorescens CECT 844 and T. melanosporum did not generally lead to a significant improvement over the positive effects of a simple inoculation of T. melanosporum; however, the addition of P. fluorescens CECT 844 did double the rate of the mycorrhization of T. melanosporum. These results may be promising for enhancing the cultivation of truffles.

  2. Growth of Amanita caesarea in the presence of Pseudomonas fluorescens and Bacillus cereus.

    Science.gov (United States)

    Cano, J M; Berrocal-Lobo, M; Domínguez-Núñez, J A

    2017-09-01

    The ectomycorrhizal (ECM) fungus Amanita caesarea CECT 20127 was tested in vitro with two potentially mycorrhizal-promoting bacterial strains, Pseudomonas fluorescens CECT 844 and Bacillus cereus CECT 148. Although P. fluorescens showed spatial and temporal compatibility with A. caesarea, it did not affect growth of the fungus. Conversely, B. cereus exhibited no such compatibility and also inhibited fungal growth. The expression pattern of the A. caesarea gene AcMST-1 was analysed using real-time quantitative polymerase chain reaction (qPCR) at three time points. This gene displays a high degree of homology with two genes, possible orthologues to AcMST-1, previously described in Amanita muscaria (AmMST-1) and Laccaria bicolor (LbMST-1) and encoding monosaccharide transporter proteins. The transcription levels of AcMST1 increased shortly after initial contact between A. caesarea and B. cereus, but expression of the gene was inhibited in the presence of P. fluorescens. Our results show that A. caesarea may possess orthologous genes of similar ECM fungal species that would allow it to adapt in nature to optimize sugar uptake from the environment depending on the presence of different microorganisms. Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.

  3. Rhizobacteria Pseudomonas fluorescens and Azospirillum sp. association enhances growth of Lactuca sativa L. under tropical conditions

    Directory of Open Access Journals (Sweden)

    Amael APONTE

    2017-06-01

    Full Text Available The selection of microorganisms that enhance plant growth and confer biotic and abiotic tolerance to crops constitutes a biotechnology currently gaining importance on a global scale. The aim of this investigation was to evaluate the effects of inoculating rhizobacteria to lettuce (Lactuca sativa L. on seed germination and vegetative development in order to use isolates as potential biofertilizers under tropical conditions. Five isolates of Pseudomonas fluorescens (Pf and one of Azospirillum sp. (Az were inoculated to seeds using a bacterial suspension of 1.5*108 CFU*mL-1. In vitro, none of the isolates promoted germination. In vivo, isolates promoted growth and acted as stress alleviators by conferring tolerance to high temperatures (≥ 30 °C. The highest seedling emergence percentages were induced by the association of P.fluorescens with Azospirillum. This association also promoted the highest leaf-area in 25 d seedlings and exhibited a significantly higher dry-weight in 40 d plants compared to the control (P≤0.05 supporting the advantages of bio-consortiums over individual strains. The strains were able to produce dependent L-tryptophan indole-3-acetic acid (IAA, to solubilize phosphorous in vitro and tolerated at least 5%-salt stress. The results indicate that isolate Pf (26 and Az possess plant growth promoting rhizobacteria (PGPR traits and should be further assessed. This study suggests that P. fluorescens and Azospirillum act synergically and are able to trigger an induced-tolerance mechanism in lettuce under abiotic stress.

  4. Ability of the marine bacterium Pseudomonas fluorescens BA3SM1 to counteract the toxicity of CdSe nanoparticles.

    Science.gov (United States)

    Poirier, Isabelle; Kuhn, Lauriane; Demortière, Arnaud; Mirvaux, Boris; Hammann, Philippe; Chicher, Johana; Caplat, Christelle; Pallud, Marie; Bertrand, Martine

    2016-10-04

    In the marine environment, bacteria from estuarine and coastal sediments are among the first targets of nanoparticle pollution; it is therefore relevant to improve the knowledge of interactions between bacteria and nanoparticles. In this work, the response of the marine bacterium Pseudomonas fluorescens BA3SM1 to CdSe nanocrystals (CdSe NPs) of 3nm (NP3) and 8nm (NP8) in diameter was evaluated through microscopic, physiological, biochemical and proteomic approaches. Transmission electron microscopy images showed that NP3 were able to penetrate the bacteria, while NP8 were highly concentrated around the cells, embedded in large exopolysaccharides. In our experimental conditions, both CdSe NP sizes induced a decrease in respiration during the stationary growth phase, while only NP8 caused growth retardation and a decrease in pyoverdine production. Proteomic analyses highlighted that the strain responded to CdSe NP toxicity by inducing various defence mechanisms such as cell aggregation, extracellular CdSe NP sequestration, effective protection against oxidative stress, modifications of envelope organization and properties, and cadmium export. In addition, BA3SM1 presented a biosorption capacity of 1.6×10(16)NP3/g dry weight and 1.7×10(15)NP8/g dry weight. This strain therefore appears as a promising agent for NP bioremediation processes. Proteomic data are available via ProteomeXchange with identifier PXD004012. To the best of our knowledge, this is the first report focussing on the effects of CdSe colloidal nanocrystals (CdSe NPs) on a marine strain of Pseudomonas fluorescens. CdSe NPs are extensively used in the industry of renewable energies and it is regrettably expected that these pollutants will sometime soon appear in the marine environment through surface runoff, urban effluents and rivers. Bacteria living in estuarine and coastal sediments will be among the first targets of these new pollutants. The pseudomonads are frequently found in these ecosystems

  5. 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.

  6. Predictive modeling of Pseudomonas fluorescens growth under different temperature and pH values

    Directory of Open Access Journals (Sweden)

    Letícia Dias dos Anjos Gonçalves

    Full Text Available Abstract Meat is one of the most perishable foods owing to its nutrient availability, high water activity, and pH around 5.6. These properties are highly conducive for microbial growth. Fresh meat, when exposed to oxygen, is subjected to the action of aerobic psychrotrophic, proteolytic, and lipolytic spoilage microorganisms, such as Pseudomonas spp. The spoilage results in the appearance of slime and off-flavor in food. In order to predict the growth of Pseudomonas fluorescens in fresh meat at different pH values, stored under refrigeration, and temperature abuse, microbial mathematical modeling was applied. The primary Baranyi and Roberts and the modified Gompertz models were fitted to the experimental data to obtain the growth parameters. The Ratkowsky extended model was used to determine the effect of pH and temperature on the growth parameter µmax. The program DMFit 3.0 was used for model adjustment and fitting. The experimental data showed good fit for both the models tested, and the primary and secondary models based on the Baranyi and Roberts models showed better validation. Thus, these models can be applied to predict the growth of P. fluorescens under the conditions tested.

  7. 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.

  8. Biocontrol of Rhizoctonia solani in native potato (Solanum phureja plants using native Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    Gloria Bautista

    2007-01-01

    Full Text Available Rhizoctonia solani is a soil borne phytopathogen associated with reduced plant vigor and tuber production in potato crops. There is a huge interest to search alternatives of biological control management of this disease, because the potato crops in Colombia are the highest consumers of chemical pesticides in Colombia. In order to obtain a fluorescent Pseudomonas strain with the capacity to reduce the disease symptoms produced by R. solani, determination and isolation of the predominant fluorescent Pseudomonas in several potato crops of the main Colombian producing region was done in a previous study. Six different P. fluorescens strains with none, moderate and high fungal growth inhibition capacity in vitro, were used in this study. Despite of the differences found in the dynamics of colonization and colonization capacity, all evaluated strains induced S. phureja growth and reduced disease symptoms produced by R. solani. Our results support the conclusion that association of P. fluorescens strains with S. phureja rhizosphere is a feasible alternative for the management of R. solani symptoms.

  9. Bioremediation of chromium contaminated soil by Pseudomonas fluorescens and indigenous microorganisms.

    Science.gov (United States)

    Jeyalakshmi, D; Kanmani, S

    2008-01-01

    Chromium is one of the toxic and hazardous pollutants in industrial wastewaters leading to soil contamination. In this study, the feasibility of remediating chromium contaminated soil using indigenous microorganisms and Pseudomonas fluorescens was evaluated. The contaminated soil sample was collected from Vellore and the pH, moisture content and chromium content were found to be 8.4, 22.5% and 5.1 mg/kg respectively. The effect of chromium on engineering properties showed decrease in permeability by 45.15%. For Pseudomonas fluorescens, the optimum pH, moisture content, biomass concentration and carbon source were found as 6.5, 20%, 10 mL and 10 mL/100 g respectively and for isolated mixed culture, the optimum parameters were found as 8.4, 25%, 15 mL and 15mL / 100 g respectively. Under optimum conditions, the reactor study showed 71.7% chromium reduction after 20 days. From the study, the bioremediation of chromium-contaminated soil by indigenous microorganisms was found to be a promising solution and after bioremediation, the engineering properties of the soil were found to be improved.

  10. Interaction between Medicago truncatula and Pseudomonas fluorescens: evaluation of costs and benefits across an elevated atmospheric CO(2.

    Directory of Open Access Journals (Sweden)

    Clémentine Lepinay

    Full Text Available Soil microorganisms play a key role in both plants nutrition and health. Their relation with plant varies from mutualism to parasitism, according to the balance of costs and benefits for the two partners of the interaction. These interactions involved the liberation of plant organic compounds via rhizodeposition. Modification of atmospheric CO(2 concentration may affect rhizodeposition and as a consequence trophic interactions that bind plants and microorganisms. Positive effect of elevated CO(2 on plants are rather well known but consequences for micoorganisms and their interactions with plants are still poorly understood. A gnotobiotic system has been developed to study the interaction between Medicago truncatula Jemalong J5 and the mutualistic bacteria Pseudomonas fluorescens strain C7R12 under two atmospheric CO(2 concentrations: ambient (365 ppm versus enriched (750 ppm. Costs and benefits for each partner have been determined over time by measuring plant development and growth, the C and N contents of the various plant parts and the density of the bacteria in rhizosphere compartments. Following the increase in CO(2, there was a beneficial effect of P. fluorescens C7R12 on development, vegetative growth, and C/N content of M. truncatula. Concerning plant reproduction, an early seed production was noticed in presence of the bacterial strain combined with increased atmospheric CO(2 conditions. Paradoxically, this transient increase in seed production was correlated with a decrease in bacterial density in the rhizosphere soil, revealing a cost of increased CO(2 for the bacterial strain. This shift of costs-benefits ratio disappeared later during the plant growth. In conclusion, the increase in CO(2 concentration modifies transiently the cost-benefit balance in favor of the plant. These results may be explained either by a competition between the two partners or a change in bacterial physiology. The ecosystem functioning depends on the

  11. Complete genome sequence of the lytic cold-active Pseudomonas fluorescens bacteriophage VSW-3 from Napahai plateau wetland.

    Science.gov (United States)

    Zhang, Chunjing; Zhang, Zhongyao; Li, Jiankai; Qin, Kunhao; Wei, Yunlin; Zhang, Qi; Lin, Lianbing; Ji, Xiuling

    2017-02-01

    The lytic cold-active bacteriophage VSW-3, belonging to the Podoviridae family and infecting the host Pseudomonas fluorescens SW-3, was isolated from the Napahai plateau wetland in China. With the development of sequencing technology, the study of Pseudomonas genomic diversity has increased; however, knowledge of cold-active phages infecting Pseudomonas is limited. The newly sequenced phage VSW-3 was classified based on virion morphology by transmission electron microscope. Sequence analysis revealed that the genome size was 40,556 bp with an overall GC content of 57.54 % and 46 open reading frames. The genome was organized into several modules containing genes for packaging, structural proteins, replication/transcription, and phage lysis. The sequence contained 45 potential promoters, 3 transcription terminators, and yet no tRNAs. This is the first report of cold-active Pseudomonas fluorescens bacteriophage genome sequencing.

  12. 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.

  13. Genetic Responses Induced in Olive Roots upon Colonization by the Biocontrol Endophytic Bacterium Pseudomonas fluorescens PICF7

    Science.gov (United States)

    Schilirò, Elisabetta; Ferrara, Massimo; Nigro, Franco; Mercado-Blanco, Jesús

    2012-01-01

    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. PMID:23144916

  14. 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.

  15. Draft Genome Sequence of the Polycyclic Aromatic Hydrocarbon-Degrading, Genetically Engineered Bioluminescent Bioreporter Pseudomonas fluorescens HK44

    Energy Technology Data Exchange (ETDEWEB)

    Chauhan, Archana [ORNL; Layton, Alice [University of Tennessee, Knoxville (UTK); Williams, Daniel W [ORNL; Smart, Abby E. [University of Tennessee, Knoxville (UTK); Ripp, Steven Anthony [ORNL; Karpinets, Tatiana V [ORNL; Brown, Steven D [ORNL; Sayler, Gary Steven [ORNL

    2011-01-01

    Pseudomonas fluorescens strain HK44 (DSM 6700) is a genetically engineered lux-based bioluminescent bioreporter. Here we report the draft genome sequence of strain HK44. Annotation of {approx}6.1 Mb sequence indicates that 30% of the traits are unique and distributed over 5 genomic islands, a prophage and two plasmids.

  16. Wave-like distribution patterns of Gfp-marked Pseudomonas fluorescens along roots of wheat plants grown in two soils

    NARCIS (Netherlands)

    Bruggen, van A.H.C.; Semenov, A.M.; Zelenev, V.V.; Semenov, A.V.; Raaijmakers, J.M.; Sayler, R.J.; Vos, de O.J.

    2008-01-01

    Culturable rhizosphere bacterial communities had been shown to exhibit wave-like distribution patterns along wheat roots. In the current work we show, for the first time, significant wave-like oscillations of an individual bacterial strain, the biocontrol agent Pseudomonas fluorescens 32 marked with

  17. Draft Genome Sequence of the Polycyclic Aromatic Hydrocarbon-Degrading, Genetically Engineered Bioluminescent Bioreporter Pseudomonas fluorescens HK44 ▿

    Science.gov (United States)

    Chauhan, Archana; Layton, Alice C.; Williams, Daniel E.; Smartt, Abby E.; Ripp, Steven; Karpinets, Tatiana V.; Brown, Steven D.; Sayler, Gary S.

    2011-01-01

    Pseudomonas fluorescens strain HK44 (DSM 6700) is a genetically engineered lux-based bioluminescent bioreporter. Here we report the draft genome sequence of strain HK44. Annotation of ∼6.1 Mb of sequence indicates that 30% of the traits are unique and distributed over five genomic islands, a prophage, and two plasmids. PMID:21742869

  18. 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...

  19. Kinetic modelling of enzyme inactivation : kinetics of heat inactivation of the extracellular proteinase from Pseudomonas fluorescens 22F

    NARCIS (Netherlands)

    Schokker, E.P.

    1997-01-01

    The kinetics of heat inactivation of the extracellular proteinase from Pseudomonas fluorescens 22F was studied. It was established, by making use of kinetic modelling, that heat inactivation in the temperature range 35 - 70 °C was most likely caused

  20. Mobile genetic elements in the genome of the beneficial rhizobacterium Pseudomonas fluorescens Pf-5.

    Science.gov (United States)

    Mavrodi, Dmitri V; Loper, Joyce E; Paulsen, Ian T; Thomashow, Linda S

    2009-01-13

    Pseudomonas fluorescens Pf-5 is a plant-associated bacterium that inhabits the rhizosphere of a wide variety of plant species and and produces secondary metabolites suppressive of fungal and oomycete plant pathogens. The Pf-5 genome is rich in features consistent with its commensal lifestyle, and its sequence has revealed attributes associated with the strain's ability to compete and survive in the dynamic and microbiologically complex rhizosphere habitat. In this study, we analyzed mobile genetic elements of the Pf-5 genome in an effort to identify determinants that might contribute to Pf-5's ability to adapt to changing environmental conditions and/or colonize new ecological niches. Sequence analyses revealed that the genome of Pf-5 is devoid of transposons and IS elements and that mobile genetic elements (MGEs) are represented by prophages and genomic islands that collectively span over 260 kb. The prophages include an F-pyocin-like prophage 01, a chimeric prophage 03, a lambdoid prophage 06, and decaying prophages 02, 04 and 05 with reduced size and/or complexity. The genomic islands are represented by a 115-kb integrative conjugative element (ICE) PFGI-1, which shares plasmid replication, recombination, and conjugative transfer genes with those from ICEs found in other Pseudomonas spp., and PFGI-2, which resembles a portion of pathogenicity islands in the genomes of the plant pathogens Pseudomonas syringae and P. viridiflava. Almost all of the MGEs in the Pf-5 genome are associated with phage-like integrase genes and are integrated into tRNA genes. Comparative analyses reveal that MGEs found in Pf-5 are subject to extensive recombination and have evolved in part via exchange of genetic material with other Pseudomonas spp. having commensal or pathogenic relationships with plants and animals. Although prophages and genomic islands from Pf-5 exhibit similarity to MGEs found in other Pseudomonas spp., they also carry a number of putative niche-specific genes that

  1. A modular esterase from Pseudomonas fluorescens subsp. cellulosa contains a non-catalytic cellulose-binding domain.

    OpenAIRE

    Ferreira, L M; Wood, T M; Williamson, G; Faulds, C; Hazlewood, G P; Black, G W; Gilbert, H J

    1993-01-01

    The 5' regions of genes xynB and xynC, coding for a xylanase and arabinofuranosidase respectively, are identical and are reiterated four times within the Pseudomonas fluorescens subsp. cellulosa genome. To isolate further copies of the reiterated xynB/C 5' region, a genomic library of Ps. fluorescens subsp. cellulosa DNA was screened with a probe constructed from the conserved region of xynB. DNA from one phage which hybridized to the probe, but not to sequences upstream or downstream of the ...

  2. Pseudomonas fluorescens

    African Journals Online (AJOL)

    Subhadfip Nandi

    2013-04-10

    Apr 10, 2013 ... Damping off caused by Sclerotium rolfsii on cowpea results in yield losses with serious socio- economic implication. Induction of defense responses by plant growth promoting rhizobacteria (PGPR) is largely associated with the production of defense enzyme phenyl ammonia lyase (PAL) and oxidative.

  3. Evolution of fitness trade-offs in locally adapted populations of Pseudomonas fluorescens

    DEFF Research Database (Denmark)

    Schick, Alana; Bailey, Susan; Kassen, Rees

    2015-01-01

    characterize the genetic causes of trade-offs generating local adaptation in populations of Pseudomonas fluorescens that had previously been evolved for specialization on three different carbon resources. We measured the fitness effects of mutations that arose during selection in that environment...... and in alternative environments to quantify the degree of specialization. We find that all mutations are beneficial in the environment of selection and that those arising later during adaptation are associated with increasingly antagonistic effects in alternative environments compared with those arising earlier......, consistent with a multioptima version of Fisher’s geometric model of adaptation. We also find that fitness of pairs of beneficial mutations are consistently less than additive in selection environments, producing a pattern of diminishing returns, but are more variable in alternative environments, being...

  4. Alginate Biosynthesis Factories in Pseudomonas fluorescens: Localization and Correlation with Alginate Production Level

    Science.gov (United States)

    Maleki, Susan; Almaas, Eivind; Zotchev, Sergey; Valla, Svein

    2015-01-01

    Pseudomonas fluorescens is able to produce the medically and industrially important exopolysaccharide alginate. The proteins involved in alginate biosynthesis and secretion form a multiprotein complex spanning the inner and outer membranes. In the present study, we developed a method by which the porin AlgE was detected by immunogold labeling and transmission electron microscopy. Localization of the AlgE protein was found to depend on the presence of other proteins in the multiprotein complex. No correlation was found between the number of alginate factories and the alginate production level, nor were the numbers of these factories affected in an algC mutant that is unable to produce the precursor needed for alginate biosynthesis. Precursor availability and growth phase thus seem to be the main determinants for the alginate production rate in our strain. Clustering analysis demonstrated that the alginate multiprotein complexes were not distributed randomly over the entire outer cell membrane surface. PMID:26655760

  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

    in central metabolism (carB) generates such a striking phenotype. We show that colony switching is underpinned by ON/OFF expression of capsules consisting of a colanic acid-like polymer. We use molecular genetics, biochemical analyses, and experimental evolution to establish that capsule switching results......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...... insight was delivered by an experiment in which populations of the bacterium Pseudomonas fluorescens SBW25 evolved, de novo, the ability to switch between two colony phenotypes. Here we unravel the molecular mechanism behind colony switching, revealing how a single nucleotide change in a gene enmeshed...

  6. Upon impact: the fate of adhering Pseudomonas fluorescens cells during nanofiltration.

    Science.gov (United States)

    Habimana, Olivier; Semião, Andrea J C; Casey, Eoin

    2014-08-19

    Nanofiltration (NF) is a high-pressure membrane filtration process increasingly applied in drinking water treatment and water reuse processes. NF typically rejects divalent salts, organic matter, and micropollutants. However, the efficiency of NF is adversely affected by membrane biofouling, during which microorganisms adhere to the membrane and proliferate to create a biofilm. Here we show that adhered Pseudomonas fluorescens cells under high permeate flux conditions are met with high fluid shear and convective fluxes at the membrane-liquid interface, resulting in their structural damage and collapse. These results were confirmed by fluorescent staining, flow cytometry, and scanning electron microscopy. This present study offers a "first-glimpse" of cell damage and death during the initial phases of bacterial adhesion to NF membranes and raises a key question about the role of this observed phenomena during early-stage biofilm formation under permeate flux and cross-flow conditions.

  7. Utilization of Pseudomonas fluorescens as Antimicrobial agent on Lipolysis Inhibitition of Rice Bran

    Directory of Open Access Journals (Sweden)

    Titin Widiyastuti

    2002-01-01

    Full Text Available Feedstuff was limited by fat content, certainly, if the material feedstuff was storage on long time periods. Fat was oxidized or hydrolyzed, which was decreased nutritional quality. A Research on Inhibited of  Lypolysis Rice Bran with anti-microbial compound of Pseudomonas fluorescens was conducted during ten month. The research used experiment methods, with Randomized Completely Block Design, storage as block and concentration of anti-mikrobial compound as treatment. Fat and polyunsaturated fatty acids content during storage was observed. Result of experiment showed anti-microbial compound inhibited oxidize and hydrolyze process of fat rice bran during six month storage. (Animal Production 4(2: 89-93 (2002

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

    Science.gov (United States)

    Gamal, Rawia F.; Abdelhady, Hemmat M.; Khodair, Taha A.; El-Tayeb, Tarek S.; Hassan, Enas A.; Aboutaleb, Khadiga A.

    2013-01-01

    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. PMID:24294253

  9. BIOCONTROL OF Rhizoctonia solani IN NATIVE POTATO (Solanum phureja PLANTS USING NATIVE Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    GLORIA BAUTISTA

    2007-01-01

    Full Text Available Rhizoctonia solani es un hongo fitopatógeno del suelo, el cual produce una reducción significativa del vigor de las plantas y de la producción de tubérculos en cultivos de papa. Es de gran interés la búsqueda de alternativas de manejo de esta enfermedad, especialmente desde la perspectiva de control biológico ya que los cultivos de papa son los mayores consumidores de plaguicidas de origen químicos en Colombia. Con el objeto de obtener una cepa del grupo de las Pseudomonas fluorescentes con la capa- cidad para reducir los síntomas de la enfermedad producidos por R. solani, se realizó en un estudio previo el aislamiento y caracterización de una colección de aislamientos de Pseudomonas fluorescentes provenientes de diferentes cultivos de la región papera más productiva del país. Seis cepas nativas de P. fluorescens con buena, moderada o ninguna capacidad para inhibir el crecimiento fúngico in vitro fueron seleccionadas. A pesar de las diferencias encontradas en términos de la dinámica y capacidad de colonización, todas las cepas evaluadas indujeron el crecimiento en las plantas de S. phureja y redujeron los síntomas de la enfermedad producidos por R. solani a nivel de invernadero. Nuestros resultados sustentan la conclusión que la asociación de cepas de P. fluorescens con la rizosfera de S. phureja es una alternativa para el manejo de R. solani en papa.

  10. LapD Is a Bis-(3', 5')-Cyclic Dimeric GMP-Binding Protein That Regulates Surface Attachment by Pseudomonas fluorescens Pf0-1

    National Research Council Canada - National Science Library

    Peter D. Newell; Russell D. Monds; George A. O'Toole; Emil C. Gotschlich

    2009-01-01

    .... For Pseudomonas fluorescens Pf0-1, c-di-GMP impacts the secretion and localization of the adhesin LapA, which is absolutely required for stable surface attachment and biofilm formation by this bacterium...

  11. 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.

  12. Investigation of zoonotic disease pathogens (Aeromonas hydrophila, Pseudomonas fluorescens, Streptococcus iniae) seen in carp farms in the Northern Iraq-Erbil region by molecular methods

    Science.gov (United States)

    Ibraheem, Azad Saber; Önalan, Şükrü; Arabacı, Muhammed

    2017-04-01

    The aim of this study was to determine the zoonotic bacteria in carp farms in the Northern Iraq-Erbil region. Carp is the main fish species cultured in Erbil region. The most common zoonotic bacteria generally seen in carp farms are Aeromonas hydrophila, Pseudomonas fluorescens and Streptococcus iniae. Samples were collected from 25 carp farms in the Northern Iraq-Erbil region. Six carp samples were collected from each carp farm. Head kidney and intestine tissue samples were collected from each carp sample. Then head kidney and intestine tissue samples were pooled separately from each carp farm. Total bacterial DNA had been extracted from the 25 pooled head kidney and 25 intestinal tissue samples. The pathogen Primers were originally designed from 16S RNA gene region. Zoonotic bacteria were scanned in all tissue samples with absent/present analysis by RT-PCR. Furthermore, the capillary gel electrophoresis bands were used for confirmation of amplicon size which was planned during primer designing stage. As a result, thirteen carp farms were positive in the respect to Aeromonas hydrophila, eight carp farms were positive from head kidney and six carp farms were positive from the intestine, only one carp farm was positive from both head kidney and the intestine tissue samples. In the respect to Streptococcus iniae, four carp farms were positive from head kidney and two carp farms were positive from the intestine. Only one carp farm was positive in the respect to Pseudomonas fluorescens from the intestine. Totally, 9 of 25 carp farms were cleared (negative) the zoonotic bacteria. In conclusion, the zoonotic bacteria were high (64 %) in carp farms in the Northern Iraq-Erbil region.

  13. KAJIAN MEKANISME ANTAGONIS PSEUDOMONAS FLUORESCENS P60 TERHADAP FUSARIUM OXYSPORUM F.SP. LYCOPERSICI PADA TANAMAN TOMAT IN VIVO

    Directory of Open Access Journals (Sweden)

    Loekas Soesanto, Endang Mugiastuti & Ruth Feti Rahayuniati .

    2011-11-01

    Full Text Available Antagonistic mechanisms study of Pseudomonas fluorescens P60 on Fusarium oxysporum f.sp. lycopersici of tomato in vivo.  This research was conducted to evaluate the effect of P. fluorescens P60 in controlling Fusarium wilt on tomato and its inhibition mechanisms. Randomized Block Design was used with four replicates and each consisted of 12 crops. The treatments tested were combination between supernatant or suspension of P. fluorescens P60 and application time, i.e., 5 days before planting, in the same time with planting, and 5 days after planting. Variables observed were phenolic compound (tannin, saponin, and glycoside, disease intensity, infection rate, late pathogen and antagonist population density, crop height, stem diameter, fresh and dry weight of roots, and fresh weight of fruit. The result showed that the application of P. fluorescens P60 either in supernatant or suspension form, could increase phenolic compound in the crop tissue, decrease the Fusarium wilt intensity on tomato as 66.00-77.88%, suppress infection rate as 73.18-79.09%, decrease late F. oxysporum f.sp. lycopersici density as 35.71%, increase the antagonist as 10 fold, increase crop height as 26.50%, improve root dry weight as 55.69%, and increase fruit weight crop-1 as 59.79%. Mechanisms of the antagonist P. fluorescens P60 in order to control the disease in the field were induced resistance, antibiosis, and plant growth promoting rhizobacteria.

  14. Effects of Pseudomonas fluorescens on the Water Parameters of Mycorrhizal and Non-Mycorrhizal Seedlings of Pinus halepensis

    Directory of Open Access Journals (Sweden)

    José A. Saiz de Omeñaca

    2013-08-01

    Full Text Available Inoculation of forest seedlings with mycorrhizal fungi and rhizobacteria can improve the morphological and physiological qualities of plants, especially those used for regeneration of arid areas. In this paper, under standard nursery conditions, Aleppo pine seedlings were inoculated with Pseudomonas fluorescens CECT 5281 rhizobacteria. Some of these seedlings were also inoculated with the ectomycorrhizal fungus Pisolithus tinctorius. Five months after the inoculations, we examined the growth, water parameters (osmotic potential at full turgor [Ψπfull], osmotic potential at zero turgor [Ψπ0], and the tissue modulus of elasticity near full turgor [Emax], mycorrhizal colonisation, and concentration of macronutrients (N, P, K, Ca and Mg in the seedlings. Subsequently, a trial was conducted to assess the root growth potential. P. fluorescens CECT 5281 decreased the cellular osmotic potential of P. halepensis seedlings but increased its elasticity. P. tinctorius + P. fluorescens caused osmotic adjustment at zero turgor and increased tissue elasticity, which improved tolerance to water stress. All inoculations improved the growth and nutrition of the seedlings but caused non-significant effects on root growth potential. The co-inoculation Pisolithus tinctorius + Pseudomonas fluorescens at the nursery may be a suitable technique for producing improved seedling material for restoration purposes.

  15. 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.

  16. The nucleotide sequence of a carboxymethylcellulase gene from Pseudomonas fluorescens subsp. cellulosa.

    Science.gov (United States)

    Hall, J; Gilbert, H J

    1988-07-01

    The complete nucleotide sequence of the gene coding for one of the carboxymethylcellulases (CMCase), expressed by Pseudomonas fluorescens subsp. cellulosa, has been determined. The structural gene consists of an open reading frame, commencing with an ATG start codon, of 2886 base pairs followed by a TAA stop codon. The gene was shown to code for a signal peptide which closely resembles the signal peptides of other secreted proteins. Unlike most Pseudomonas genes, the CMCase sequence does not have a high G + C (51%) content and there is no marked preference for codons ending in G or C. Upstream of the structural gene there are no sequences which bear a strong resemblance to consensus Escherichia coli promoters. A sequence is present, however, which exhibits homology to the consensus DNA sequence that binds the catabolic activator protein (CAP). Bal31 deletions of the structural gene revealed the extent by which the gene could be modified and still encode a functional CMCase. Subclones of the cellulase gene have been constructed in pUC18 and pUC19. One of the resultant plasmids, pJHS1 directs a 20-fold increase in CMCase synthesis, when compared to the original construct, pJHH2. Analysis of cells harbouring pJHS1 showed the cellulase polypeptide to have a molecular weight of 106000. This is in close agreement with the predicted size of the enzyme deduced from the nucleotide sequence data.

  17. 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...... by solubilization temperature. An antiserum against Psi1 recognized a protein of M,55,000 in four other P. fluorescens strains among 24 tested strains representing Pseudomonas rRNA homology group I, showing antigenicheterogeneity within this group. A method for immunofluorescence microscopy involving cell...

  18. 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

    of P.fluorescens AH2 was studied under iron-rich and iron-limited conditions. Sterile-filtered culture supernatants from iron- limited P. fluorescens AH2 inhibited the growth of V. anguillarum, whereas sterile-filtered supernatants from iron- replete cultures of P. fluorescens AH2 did not. P...

  19. Optimization of worm-bed leachate for culturing of tomato (Lycopersicon esculentum Mill) inoculated with Glomus fasciculatum and Pseudomonas fluorescens

    OpenAIRE

    Oliva-Llaven,María Ángela; Rodríguez-Hernández,Ludwi; Mendoza-Nazar,Paula; Ruiz-Sesma, Benigno; Álvarez-Solís,José David; Dendooven, Luc; Federico A. Gutiérrez-Miceli

    2010-01-01

    A response surface technique was used to analyze the effect of Glomus fasciculatum, Pseudomonas fluorescens and worm-bed leachate (WBL) on growth, yield and characteristics of tomato (Lycopersicon esculentum Mill). The treatments combined inoculation with or without P. fluorescensor G. fasciculatum and the application of WBL at 20% (v/v) each day or every three days. Plant height, number of leaves and yield of tomato fruits was not affected by the factors studied. However, plants with foliar ...

  20. Draft genome sequence of the polycyclic aromatic hydrocarbon-degrading, genetically engineered bioluminescent bioreporter Pseudomonas fluorescens HK44.

    Science.gov (United States)

    Chauhan, Archana; Layton, Alice C; Williams, Daniel E; Smartt, Abby E; Ripp, Steven; Karpinets, Tatiana V; Brown, Steven D; Sayler, Gary S

    2011-09-01

    Pseudomonas fluorescens strain HK44 (DSM 6700) is a genetically engineered lux-based bioluminescent bioreporter. Here we report the draft genome sequence of strain HK44. Annotation of ∼6.1 Mb of sequence indicates that 30% of the traits are unique and distributed over five genomic islands, a prophage, and two plasmids. Copyright © 2011, American Society for Microbiology. All Rights Reserved.

  1. 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.

  2. The cellodextrinase from Pseudomonas fluorescens subsp. cellulosa consists of multiple functional domains.

    Science.gov (United States)

    Ferreira, L M; Hazlewood, G P; Barker, P J; Gilbert, H J

    1991-11-01

    A genomic library of Pseudomonas fluorescens subsp. cellulosa DNA was constructed in pUC18 and Escherichia coli recombinants expressing 4-methylumbelliferyl beta-D-cellobioside-hydrolysing activity (MUCase) were isolated. Enzyme produced by MUCase-positive clones did not hydrolyse either cellobiose or cellotriose but converted cellotetraose into cellobiose and cleaved cellopentaose and cellohexaose, producing a mixture of cellobiose and cellotriose. There was no activity against CM-cellulose, insoluble cellulose or xylan. On this basis, the enzyme is identified as an endo-acting cellodextrinase and is designated cellodextrinase C (CELC). Nucleotide sequencing of the gene (celC) which directs the synthesis of CELC revealed an open reading frame of 2153 bp, encoding a protein of Mr 80,189. The deduced primary sequence of CELC was confirmed by the Mr of purified CELC (77,000) and by the experimentally determined N-terminus of the enzyme which was identical with residues 38-47 of the translated sequence. The N-terminal region of CELC showed strong homology with endoglucanase, xylanases and an arabinofuranosidase of Ps. fluorescens subsp. cellulosa; homologous sequences included highly conserved serine-rich regions. Full-length CELC bound tightly to crystalline cellulose. Truncated forms of celC from which the DNA sequence encoding the conserved domain had been deleted, directed the synthesis of a functional cellodextrinase that did not bind to crystalline cellulose. This is consistent with the N-terminal region of CELC comprising a non-catalytic cellulose-binding domain which is distinct from the catalytic domain. The role of the cellulose-binding region is discussed.

  3. Pseudomonas fluorescens F113 can produce a second flagellar apparatus, which is important for root colonization.

    Directory of Open Access Journals (Sweden)

    Emma Barahona

    2016-09-01

    Full Text Available The genomic sequence of Pseudomonas fluorescens F113 has shown the presence of a 41 kb cluster of genes that encode the production of a second flagellar apparatus. Among 2535 pseudomonads strains with sequenced genomes, these genes are only present in the genomes of F113 and other six strains, all but one belonging to the P. fluorescens cluster of species, in the form of a genetic island. The genes are homologous to the flagellar genes of the soil bacterium Azotobacter vinelandii. Regulation of these genes is mediated by the flhDC master operon, instead of the typical regulation in pseudomonads, which is through fleQ. Under laboratory conditions, F113 does not produce this flagellum and the flhDC operon is not expressed. However, ectopic expression of the flhDC operon is enough for its production, resulting in a hypermotile strain. This flagellum is also produced under laboratory conditions by the kinB and algU mutants. Genetic analysis has shown that kinB strongly represses the expression of the flhDC operon. This operon is activated by the Vfr protein probably in a c-AMP dependent way. The strains producing this second flagellum are all hypermotile and present a tuft of polar flagella instead of the single polar flagellum produced by the wild-type strain. Phenotypic variants isolated from the rhizosphere produce this flagellum and mutation of the genes encoding it, results in a defect in competitive colonization, showing its importance for root colonization.

  4. 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.

  5. Development and validation of a real-time TaqMan assay for the detection and enumeration of Pseudomonas fluorescens ATCC 13525 used as a challenge organism in testing of food equipments.

    Science.gov (United States)

    Saha, Ratul; Bestervelt, Lorelle L; Donofrio, Robert S

    2012-02-01

    Pseudomonas fluorescens ATCC 13525 is used as the challenge organism to evaluate the efficacy of the clean-in-place (CIP) process of food equipment (automatic ice-maker) as per NSF/ANSI Standard 12. Traditional culturing methodology is presently used to determine the concentration of the challenge organism, which takes 48 h to confirm the cell density. Storage of the challenge preparation in the refrigerator might alter the cell density as P. fluorescens is capable of growing at 4 °C. Also, background organism can grow on the Pseudomonas F agar (PFA) used for the recovery of P. fluorescens thus affecting the results of the test. Real-time TaqMan assay targeting the cpn60 gene was developed for the enumeration and the identification of P. fluorescens because of its specificity, accuracy, and shorter turnaround time. The TaqMan primer-probe pair developed using the Allele ID® 7.0 probe design software was highly specific and sensitive for the target organism. The sensitivity of the assay was 10 colony forming units (CFU)/mL. The assay was also successful in determining the concentration of the challenge preparation within 2 h. Based on these observations, TaqMan assay targeting the cpn60 gene can be efficiently used for strain level identification and enumeration of bacteria. Pseudomonas fluorescens ATCC 13525 is used as a challenge organism in the efficacy testing of clean-in-place process of food equipments. Currently, culturing technique is used for its identification and estimation, which is not only time-consuming but also prone to error. Real-time TaqMan assay is more specific, sensitive, and accurate along with a shorter turnaround time compared to culturing techniques, thereby increasing the overall quality of the testing methodology to evaluate the clean-in-place process critical for the food industry to protect public health and safety. © 2012 Institute of Food Technologists®

  6. 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.

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

    Science.gov (United States)

    Großkinsky, Dominik K; Tafner, Richard; Moreno, María V; Stenglein, Sebastian A; García de Salamone, Inés E; Nelson, Louise M; Novák, Ondřej; Strnad, Miroslav; van der Graaff, Eric; Roitsch, Thomas

    2016-03-17

    Plant beneficial microbes mediate biocontrol of diseases by interfering with pathogens or via strengthening the host. Although phytohormones, including cytokinins, are known to regulate plant development and physiology as well as plant immunity, their production by microorganisms has not been considered as a biocontrol mechanism. Here we identify the ability of Pseudomonas fluorescens G20-18 to efficiently control P. syringae infection in Arabidopsis, allowing maintenance of tissue integrity and ultimately biomass yield. Microbial cytokinin production was identified as a key determinant for this biocontrol effect on the hemibiotrophic bacterial pathogen. While cytokinin-deficient loss-of-function mutants of G20-18 exhibit impaired biocontrol, functional complementation with cytokinin biosynthetic genes restores cytokinin-mediated biocontrol, which is correlated with differential cytokinin levels in planta. Arabidopsis mutant analyses revealed the necessity of functional plant cytokinin perception and salicylic acid-dependent defence signalling for this biocontrol mechanism. These results demonstrate microbial cytokinin production as a novel microbe-based, hormone-mediated concept of biocontrol. This mechanism provides a basis to potentially develop novel, integrated plant protection strategies combining promotion of growth, a favourable physiological status and activation of fine-tuned direct defence and abiotic stress resilience.

  8. 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.

  9. 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.

  10. Metabolic and transcriptomic changes induced in Arabidopsis by the rhizobacterium Pseudomonas fluorescens SS101.

    Science.gov (United States)

    van de Mortel, Judith E; de Vos, Ric C H; Dekkers, Ester; Pineda, Ana; Guillod, Leandre; Bouwmeester, Klaas; van Loon, Joop J A; Dicke, Marcel; Raaijmakers, Jos M

    2012-12-01

    Systemic resistance induced in plants by nonpathogenic rhizobacteria is typically effective against multiple pathogens. Here, we show that root-colonizing Pseudomonas fluorescens strain SS101 (Pf.SS101) enhanced resistance in Arabidopsis (Arabidopsis thaliana) against several bacterial pathogens, including Pseudomonas syringae pv tomato (Pst) and the insect pest Spodoptera exigua. Transcriptomic analysis and bioassays with specific Arabidopsis mutants revealed that, unlike many other rhizobacteria, the Pf.SS101-induced resistance response to Pst is dependent on salicylic acid signaling and not on jasmonic acid and ethylene signaling. Genome-wide transcriptomic and untargeted metabolomic analyses showed that in roots and leaves of Arabidopsis plants treated with Pf.SS101, approximately 1,910 genes and 50 metabolites were differentially regulated relative to untreated plants. Integration of both sets of "omics" data pointed to a prominent role of camalexin and glucosinolates in the Pf.SS101-induced resistance response. Subsequent bioassays with seven Arabidopsis mutants (myb51, cyp79B2cyp79B3, cyp81F2, pen2, cyp71A12, cyp71A13, and myb28myb29) disrupted in the biosynthesis pathways for these plant secondary metabolites showed that camalexin and glucosinolates are indeed required for the induction of Pst resistance by Pf.SS101. Also for the insect S. exigua, the indolic glucosinolates appeared to play a role in the Pf.SS101-induced resistance response. This study provides, to our knowledge for the first time, insight into the substantial biochemical and temporal transcriptional changes in Arabidopsis associated with the salicylic acid-dependent resistance response induced by specific rhizobacteria.

  11. Characterization and expression in Escherichia coli of an endoglucanase gene of Pseudomonas fluorescens subsp. cellulosa.

    Science.gov (United States)

    Lejeune, A; Dartois, V; Colson, C

    1988-07-13

    An endoglucanase gene of Pseudomonas fluorescens subsp. cellulosa present on plasmid pRUCL150 and expressed in Escherichia coli was subcloned in plasmid pBR322. Plasmid pRUCL153 contained the smallest DNA insert (2.9 kb) with endoglucanase activity. The plasmids directed the synthesis of a mostly periplasmic enzyme in E. coli and the level of enzyme activity was comparable in several strains. Analysis by non-denaturing polyacrylamide gel electrophoresis of the endoglucanase produced with various recombinant plasmids showed that it was unique. The endoglucanase gene on plasmid pRUCL153 was localized by physical mapping of independent transposon Tn5 insertions. Hence, its size was estimated to be approx. 1.3 kb. In vivo radioactive labelling of plasmid-encoded proteins using minicells, followed by denaturing polyacrylamide gel electrophoresis, allowed us to determine the size of the endoglucanase: Mr 40,000 for the precursor and Mr 38,000 for the mature enzyme. It was demonstrated that no cellulase operon, but a single gene, was cloned. The direction of transcription of the gene was determined by placing it under the control of the promoter of the lactose operon.

  12. Influence of mineral amendment on disease suppressive activity of Pseudomonas fluorescens to Fusarium wilt of chickpea.

    Science.gov (United States)

    Saikia, Ratul; Varghese, Saju; Singh, Bhim Pratap; Arora, Dilip K

    2009-01-01

    Fusarium wilt caused by Fusarium oxysporum f. sp. ciceri causes considerable yield loss of chickpea. Pseudomonas fluorescens4-92 (Pf4-92) strain can suppress the disease. Amendment of zinc EDTA and copper EDTA could not suppress the disease significantly when used alone; however, they significantly suppressed the disease in presence of Pf4-92. In vitro observation showed that at 40, 30 and 20microgml(-1) concentrations of these minerals, i.e. Zn, Cu and Zn plus Cu, respectively, completely repressed the production of the phytotoxin, fusaric acid (FA). FA concentration (0.5microgml(-1)) has been shown to suppress the production of 2,4-diacetylphloroglucinol (DAPG) by Pf4-92, and DAPG, salicylic acid, pyochelin and pyoluteorin production was enhanced by these mineral amendments. In rockwool bioassays, Zn, Cu and Zn plus Cu amendments reduced FA production and enhanced DAPG production. This study demonstrates that Zn and Cu enhance biocontrol activity by reducing FA produced by the pathogen, F. oxysporum f. sp. ciceri.

  13. Pseudomonas fluorescens R68 assisted enhancement in growth and fertilizer utilization of Amaranthus tricolor (L.).

    Science.gov (United States)

    Jimtha John, C; Jishma, P; Karthika, N R; Nidheesh, K S; Ray, J G; Mathew, Jyothis; Radhakrishnan, E K

    2017-08-01

    Plant probiotic potential of rhizosphere microbiome and its role in phytofertilizer mobilization are largely unexplored. In the current study, the rhizobacterium Pseudomonas fluorescens R68 (PFR68) isolated from Western Ghat was analyzed for its growth enhancement effect on the leafy vegetable Amaranthus tricolor (L.). One month of field growth of PFR68 inoculated A. tricolor has found to have enhanced growth parameters such as leaf number (1.57 fold), root number (1.76 fold), shoot length (1.28 fold) and fresh weight (2.31 fold). The treatment also improved soil fertility in terms of Nitrogen, Phosphorus and Potassium content. Most remarkably, application of PFR68 alone and 50% of recommended NPK dose along with PFR68 has resulted in enhanced growth of A. tricolor comparable to plants treated with full dose of NPK. In addition to this, application of PFR68 along with 50% NPK augmented the available Nitrogen and Phosphorus content in soil. This indicates the potential of selected organism in enrichment of soil health and enhancement of crop productivity. In conclusion, field performance of PFR68 on growth of A. tricolor confirms its promises to develop into plant probiotic formulation.

  14. Correlation of Carbohydrate Catabolism and Synthesis of Macromolecules During Enzyme Synthesis in Pseudomonas fluorescens.

    Science.gov (United States)

    Kirkland, J J; Durham, N N

    1965-07-01

    Kirkland, Jerry J. (Oklahoma State University, Stillwater), and Norman N. Durham. Correlation of carbohydrate catabolism and synthesis of macromolecules during enzyme synthesis in Pseudomonas fluorescens. J. Bacteriol. 90: 23-28. 1965.-Glucose, ribose, and fructose shorten the lag period required for synthesis of protocatechuate oxygenase. Radioactivity from uracil-2-C(14) is incorporated into the hot trichloroacetic acid-soluble fraction after a lag period of approximately 20 min after addition of protocatechuic acid. Addition of glucose or ribose simultaneously with the inducer shortens the lag period to approximately 5 min and increases the rate of uracil incorporation. The inducer must be present to initiate incorporation of radioactivity, and the exogenous carbon source accelerates incorporation but is not sufficient to initiate synthesis by itself. The addition of protocatechuic acid increases the rate and total incorporation of radioactivity from uniformly labeled glucose or ribose-1-C(14) into the hot trichloroacetic acid-soluble fraction. Ribose decreases the incorporation of radioactivity from uniformly labeled glucose into the hot trichloroacetic acid-soluble fraction, and glucose shows a similar effect on incorporation of radioactivity from ribose-1-C(14), indicating the two sugars are serving in the same capacity to enhance enzyme synthesis. Radioactivity from glucose-1-C(14) is not incorporated into the hot trichloroacetic acid-soluble fraction. The results suggest that glucose and ribose shorten the lag period for inducible enzyme formation by serving as a "specific" carbon source for synthesis of macromolecules such as ribonucleic acid.

  15. Adhesion of Pseudomonas fluorescens biofilms to glass, stainless steel and cellulose.

    Science.gov (United States)

    Wan Dagang, W R Z; Bowen, J; O'Keeffe, J; Robbins, P T; Zhang, Z

    2016-05-01

    The adhesion of colloidal probes of stainless steel, glass and cellulose to Pseudomonas fluorescens biofilms was examined using atomic force microscopy (AFM) to allow comparisons between surfaces to which biofilms might adhere. Biofilm was grown on a stainless steel substrate and covered most of the surface after 96 h. AFM approach and retraction curves were obtained when the biofilm was immersed in a tryptone/soy medium. On approach, all the colloidal probes experienced a long non-contact phase more than 100 nm in length, possibly due to the steric repulsion by extracellular polymers from the biofilm and hydrophobic effects. Retraction data showed that the adhesion varied from position to position on the biofilm. The mean value of adhesion of glass to the biofilm (48 ± 7 nN) was the greatest, followed by stainless steel (30 ± 7 nN) and cellulose (7.8 ± 0.4 nN). The method allows understanding of adhesion between the three materials and biofilm, and development of a better strategy to remove the biofilm from these surfaces relevant to different industrial applications.

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

    Science.gov (United States)

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

    2015-05-29

    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 SS101 (Pf.SS101) have not been precisely elucidated. The effects of Pf.SS101 and its VOCs on augmentation of plant growth promotion were investigated in vitro and in planta. A significant growth promotion was observed in plants exposed Pf.SS101 under both conditions, suggesting that its VOCs play a key role in promoting plant growth. Solid-phase micro-extraction (SPME) and a gas chromatography-mass spectrophotometer (GC-MS) system were used to characterize the VOCs emitted by Pf.SS101 and 11 different compounds were detected in samples inoculated this bacterium, including 13-Tetradecadien-1-ol, 2-butanone and 2-Methyl-n-1-tridecene. Application of these compounds resulted in enhanced plant growth. This study suggests that Pf.SS101 promotes the growth of plants via the release of VOCs including 13-Tetradecadien-1-ol, 2-butanone and 2-Methyl-n-1-tridecene, thus increasing understanding of the role of VOCs in plant-bacterial inter-communication. Copyright © 2015 Elsevier Inc. All rights reserved.

  17. Small RNA-dependent expression of secondary metabolism is controlled by Krebs cycle function in Pseudomonas fluorescens.

    Science.gov (United States)

    Takeuchi, Kasumi; Kiefer, Patrick; Reimmann, Cornelia; Keel, Christoph; Dubuis, Christophe; Rolli, Joëlle; Vorholt, Julia A; Haas, Dieter

    2009-12-11

    Pseudomonas fluorescens CHA0, an antagonist of phytopathogenic fungi in the rhizosphere of crop plants, elaborates and excretes several secondary metabolites with antibiotic properties. Their synthesis depends on three small RNAs (RsmX, RsmY, and RsmZ), whose expression is positively controlled by the GacS-GacA two-component system at high cell population densities. To find regulatory links between primary and secondary metabolism in P. fluorescens and in the related species Pseudomonas aeruginosa, we searched for null mutations that affected central carbon metabolism as well as the expression of rsmY-gfp and rsmZ-gfp reporter constructs but without slowing down the growth rate in rich media. Mutation in the pycAB genes (for pyruvate carboxylase) led to down-regulation of rsmXYZ and secondary metabolism, whereas mutation in fumA (for a fumarase isoenzyme) resulted in up-regulation of the three small RNAs and secondary metabolism in the absence of detectable nutrient limitation. These effects required the GacS sensor kinase but not the accessory sensors RetS and LadS. An analysis of intracellular metabolites in P. fluorescens revealed a strong positive correlation between small RNA expression and the pools of 2-oxoglutarate, succinate, and fumarate. We conclude that Krebs cycle intermediates (already known to control GacA-dependent virulence factors in P. aeruginosa) exert a critical trigger function in secondary metabolism via the expression of GacA-dependent small RNAs.

  18. Characterization of a Pseudomonas fluorescens strain from tomato rhizosphere and its use for integrated management of tomato damping-off

    Energy Technology Data Exchange (ETDEWEB)

    Jayaraj, J; Parthasarathi, T.; Radhakrishnan, N.V. [Annamalai University, Annamalainagar (India). Faculty of Agriculture

    2007-10-15

    A highly antagonistic Pseudomonas fluorescens strain was isolated from tomato rhizosphere and characterized for its in vitro and in vivo biocontrol potential against Pythium aphanidermatum. The identified Pseudomonas fluorescens strain (PfT-8) was capable of producing high levels of chitinase, beta-1,3-glucanase, cellulase, fungitoxic metabolites and siderophores. Seven different carrier formulations including a talc-based powder, lignite-based powder, peat-based powder, lignite + fly ash-based powder, wettable powder, bentonite-paste and polyethylene glycol (PEG) paste were prepared utilizing PfT-8. Shelf life was evaluated for up to 6 months of storage at ambient room temperature (28{sup o}C). Biocontrol efficacy of formulations was studied under greenhouse and field conditions. Among the formulations, peat, lignite, lignite+fly-ash and bentonite paste based formulations maintained higher propagule number than others and also showed greater biocontrol potential. However, propagule number gradually decreased with time. Soil incorporation of organic amendments and specifically poultry manure and FYM, significantly reduced damping-off incidence and also augmented the rhizosphere population of the marked P. fluorescens strain that was resistant to streptomycin and rifampicin.

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

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    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.

  20. Pseudomonas fluorescens HK44: lessons learned from a model whole-cell bioreporter with a broad application history.

    Science.gov (United States)

    Trögl, Josef; Chauhan, Archana; Ripp, Steven; Layton, Alice C; Kuncová, Gabriela; Sayler, Gary S

    2012-01-01

    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.

  1. 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.

  2. Expression of the translocator protein (TSPO from Pseudomonas fluorescens Pfo-1 requires the stress regulatory sigma factors AlgU and RpoH

    Directory of Open Access Journals (Sweden)

    Charlène eLeneveu-Jenvrin

    2015-09-01

    Full Text Available The translocator protein (TSPO, previously designated as peripheral-type benzodiazepine receptor, is an evolutionary conserved protein that is found in many Eukarya, Archae and Bacteria, in which it plays several important functions including for example membrane biogenesis, signaling and stress response. A tspo homologue gene has been identified in several members of the Pseudomonas genus, among which the soil bacterium P. fluorescens Pf0-1. In this bacterium, the tspo gene is located in the vicinity of a putative hybrid histidine kinase-encoding gene. Since tspo has been involved in water stress related response in plants, we explored the effects of hyperosmolarity and temperature on P. fluorescens Pf0-1 tspo expression using a strategy based on lux-reporter fusions. We show that the two genes Pfl01_2810 and tspo are co-transcribed forming a transcription unit. The expression of this operon is growth phase-dependent and is increased in response to high concentrations of NaCl, sucrose and to a D-cycloserine treatment, which are conditions leading to activity of the major cell wall stress responsive extracytoplasmic sigma factor AlgU. Interestingly, the promoter region activity is strongly lowered in a P. aeruginosa algU mutant, suggesting that AlgU may be involved at least partly in the molecular mechanism leading to Pfl01_2810-tspo expression. In silico analysis of this promoter region failed to detect an AlgU consensus binding site; however, we detect a putative binding site for the heat shock response RpoH sigma factor. Accordingly, the promoter activity of the region containing this sequence is increased in response to high growth temperature and slightly lowered in a P. aeruginosa rpoH mutant strain. Taken together, our data suggest that P. fluorescens tspo gene may belong at least partly to the cell wall stress response.

  3. Control biológico de Rhizoctonia solani en plantas de papa criollaSolanum phureja usando cepas nativas de Pseudomonas fluorescens BIOCONTROL OF Rhizoctonia solani IN NATIVE POTATO (Solanum phureja PLANTS USING NATIVE Pseudomonas fluorescens

    Directory of Open Access Journals (Sweden)

    GLORIA BAUTISTA

    2007-06-01

    Full Text Available Rhizoctonia solani es un hongo fitopatógeno del suelo, el cual produce una reducción significativa del vigor de las plantas y de la producción de tubérculos en cultivos de papa. Es de gran interés la búsqueda de alternativas de manejo de esta enfermedad, especialmente desde la perspectiva de control biológico ya que los cultivos de papa son los mayores consumidores de plaguicidas de origen químicos en Colombia. Con el objeto de obtener una cepa del grupo de las Pseudomonas fluorescentes con la capacidad para reducir los síntomas de la enfermedad producidos por R. solani, se realizó en un estudio previo el aislamiento y caracterización de una colección de aislamientos de Pseudomonas fluorescentes provenientes de diferentes cultivos de la región papera más productiva del país. Seis cepas nativas de P. fluorescens con buena, moderada o ninguna capacidad para inhibir el crecimiento fúngico in vitro fueron seleccionadas. A pesar de las diferencias encontradas en términos de la dinámica y capacidad de colonización, todas las cepas evaluadas indujeron el crecimiento en las plantas de S. phureja y redujeron los síntomas de la enfermedad producidos por R. solani a nivel de invernadero. Nuestros resultados sustentan la conclusión que la asociación de cepas de P. fluorescens con la rizosfera de S. phureja es una alternativa para el manejo de R. solani en papa.Rhizoctonia solani is a soil borne phytopathogen associated with reduced plant vigor and tuber production in potato crops. There is a huge interest to search alternatives of biological control management of this disease, because the potato crops in Colombia are the highest consumers of chemical pesticides in Colombia. In order to obtain a fluorescent Pseudomonas strain with the capacity to reduce the disease symptoms produced by R. solani, determination and isolation of the predominant fluorescent Pseudomonas in several potato crops of the main Colombian producing region was done

  4. Pemanfaatan Beberapa Kaldu Hewan sebagai Bahan Formula Cair Pseudomonas fluorescens P60 untuk Mengendalikan Sclerotium rolfsii pada Tanaman Mentimun

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    Endang Mugiastuti

    2011-07-01

    Full Text Available A research aiming at knowing the potency of several animal broths as organic liquid formula of Pseudomonas fluorescens P60, soaking period of Sclerotium rolfsii sclerotia, and its application method on cucumber stem-end rot was done. Completely randomized design and randomized block design both arranged by factorial were used for in vitro and in planta tests, respectively. The first factor was six kinds of animal broth, i.e., golden snail, local chicken,broiler chicken, catfish, cow bone, and rat. The second one for in vitro test was the soaking period in the Pseudomonas fluorescens P60 formula, i.e., 0, 1, 10, and 100 hours and for in planta one was application methods, i.e., seed soaking or crop spraying. Result of the research showed that the best animal broth as liquid formula for Pseudomonas fluorescens P60 was golden snail broth indicated by suppression of sclerotial germination up to 97.4% after soaking for 100 hours. The best application method to suppress the disease was spraying method showed by suppressed of sclerotial germination, longer incubation period, and suppressed disease incidence and sclerotial late population of 55.79, 147.35, 66.67, and 59.68%, repectively. Spraying the formula could also increase crop height difference, fresh and dry weight of crop, fresh and dry weight of root, and root length to 146.83, 86.62, 112.5, 87.88, 140, and 159.68%, respectively. Penelitian dengan tujuan untuk mengetahui potensi beberapa kaldu hewan sebagai formula cair organik Pseudomonas fluorescens P60, lama perendaman sklerotium Sclerotium rolfsii, dan cara aplikasinya terhadap penyakit busuk pangkal batang mentimun dilakukan. Rancangan acak lengkap faktorial dan rancangan acak kelompok faktorial digunakan dalam penelitian in vitro dan in planta. Faktor pertama yang diuji adalah enam jenis kaldu yang terdiri atas kaldu keong mas, ayam kampung, ayam potong, ikan lele, tulang sapi, dan tikus. Faktor kedua pada penelitian in vitro adalah lama

  5. A comparison between Pseudomonas aureofaciens (chlororaphis and P. fluorescens in biological control of cotton seedling damping-off disease

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    Samaneh Samavat

    2014-07-01

    Full Text Available Due to the importance of the biological control of plant diseases, testing and introducing new biocontrol-active microorganisms is a major concern among plant pathologists. The causal agent of cotton seedling damping-off disease is Rhizoctonia solani. In this regard, we tried to investigate the antagonistic activities of Pseudomonas aureofaciens (chlororaphis 30–84 (phenazine producing wild type and non-phenazine producing mutant strains on R. solani, in comparison with some isolates of P. fluorescent under both in vitro (laboratory and in vivo (greenhouse conditions. In the laboratory experiment, the inhibitory effects of all the bacteria, on the growth of R. solani, were evaluated using the dual culture procedure. Results showed that five isolates of P. fluorescent along with both strains of P. aureofaciens significantly inhibited the growth of R. solani. Effective bacterial antagonists were then evaluated in a greenhouse experiment where cotton seeds were coated with their suspensions and were sown in pasteurised field-soil. The soil had been pre-inoculated with a virulent isolate of R. solani. The efficacy of the bacterial antagonists was evaluated by counting the number of surviving seedlings in different treatments, at 15 and 60 days after sowing, for determining pre- and post-emergence damping-off incidence. According to the results of the greenhouse experiment, at both intervals, two isolates of P. fluorescens along with both strains of P. aureofaciens caused significant increases in the number of healthy seedlings, in comparison with the untreated control, and a commonly used fungicide (carboxin-thiram. The efficacy of phenazine producing a wild type strain of P. aureofaciens was higher than its non-phenazine producing mutant, indicating that phenazine plays an important role in the antagonistic activity of P. aureofaciens. Effective bacterial antagonists were then studied for their antagonistic mechanisms. The results showed that all

  6. Pyrroloquinoline Quinone Is a Plant Growth Promotion Factor Produced by Pseudomonas fluorescens B161

    Science.gov (United States)

    Choi, Okhee; Kim, Jinwoo; Kim, Jung-Gun; Jeong, Yeonhwa; Moon, Jae Sun; Park, Chang Seuk; Hwang, Ingyu

    2008-01-01

    Pseudomonas fluorescens B16 is a plant growth-promoting rhizobacterium. To determine the factors involved in plant growth promotion by this organism, we mutagenized wild-type strain B16 using ΩKm elements and isolated one mutant, K818, which is defective in plant growth promotion, in a rockwool culture system. A cosmid clone, pOK40, which complements the mutant K818, was isolated from a genomic library of the parent strain. Tn3-gusA mutagenesis of pOK40 revealed that the genes responsible for plant growth promotion reside in a 13.3-kb BamHI fragment. Analysis of the DNA sequence of the fragment identified 11 putative open reading frames, consisting of seven known and four previously unidentified pyrroloquinoline quinone (PQQ) biosynthetic genes. All of the pqq genes showed expression only in nutrient-limiting conditions in a PqqH-dependent manner. Electrospray ionization-mass spectrometry analysis of culture filtrates confirmed that wild-type B16 produces PQQ, whereas mutants defective in plant growth promotion do not. Application of wild-type B16 on tomato (Solanum lycopersicum) plants cultivated in a hydroponic culture system significantly increased the height, flower number, fruit number, and total fruit weight, whereas none of the strains that did not produce PQQ promoted tomato growth. Furthermore, 5 to 1,000 nm of synthetic PQQ conferred a significant increase in the fresh weight of cucumber (Cucumis sativus) seedlings, confirming that PQQ is a plant growth promotion factor. Treatment of cucumber leaf discs with PQQ and wild-type B16 resulted in the scavenging of reactive oxygen species and hydrogen peroxide, suggesting that PQQ acts as an antioxidant in plants. PMID:18055583

  7. Pyrroloquinoline quinone is a plant growth promotion factor produced by Pseudomonas fluorescens B16.

    Science.gov (United States)

    Choi, Okhee; Kim, Jinwoo; Kim, Jung-Gun; Jeong, Yeonhwa; Moon, Jae Sun; Park, Chang Seuk; Hwang, Ingyu

    2008-02-01

    Pseudomonas fluorescens B16 is a plant growth-promoting rhizobacterium. To determine the factors involved in plant growth promotion by this organism, we mutagenized wild-type strain B16 using OmegaKm elements and isolated one mutant, K818, which is defective in plant growth promotion, in a rockwool culture system. A cosmid clone, pOK40, which complements the mutant K818, was isolated from a genomic library of the parent strain. Tn3-gusA mutagenesis of pOK40 revealed that the genes responsible for plant growth promotion reside in a 13.3-kb BamHI fragment. Analysis of the DNA sequence of the fragment identified 11 putative open reading frames, consisting of seven known and four previously unidentified pyrroloquinoline quinone (PQQ) biosynthetic genes. All of the pqq genes showed expression only in nutrient-limiting conditions in a PqqH-dependent manner. Electrospray ionization-mass spectrometry analysis of culture filtrates confirmed that wild-type B16 produces PQQ, whereas mutants defective in plant growth promotion do not. Application of wild-type B16 on tomato (Solanum lycopersicum) plants cultivated in a hydroponic culture system significantly increased the height, flower number, fruit number, and total fruit weight, whereas none of the strains that did not produce PQQ promoted tomato growth. Furthermore, 5 to 1,000 nm of synthetic PQQ conferred a significant increase in the fresh weight of cucumber (Cucumis sativus) seedlings, confirming that PQQ is a plant growth promotion factor. Treatment of cucumber leaf discs with PQQ and wild-type B16 resulted in the scavenging of reactive oxygen species and hydrogen peroxide, suggesting that PQQ acts as an antioxidant in plants.

  8. Cost of Adaptation and Fitness Effects of Beneficial Mutations in Pseudomonas fluorescens

    Science.gov (United States)

    Bataillon, Thomas; Zhang, Tianyi; Kassen, Rees

    2011-01-01

    Adaptations are constructed through the sequential substitution of beneficial mutations by natural selection. However, the rarity of beneficial mutations has precluded efforts to describe even their most basic properties. Do beneficial mutations typically confer small or large fitness gains? Are their fitness effects environment specific, or are they broadly beneficial across a range of environments? To answer these questions, we used two subsets (n = 18 and n = 63) of a large library of mutants carrying antibiotic resistance mutations in the bacterium Pseudomonas fluorescens whose fitness, along with the antibiotic sensitive ancestor, was assayed across 95 novel environments differing in the carbon source available for growth. We explore patterns of genotype-by-environment (G×E) interactions and ecological specialization among the 18 mutants initially found superior to the sensitive ancestor in one environment. We find that G×E is remarkably similar between the two sets of mutants and that beneficial mutants are not typically associated with large costs of adaptation. Fitness effects among beneficial mutants depart from a strict exponential distribution: they assume a variety of shapes that are often roughly L shaped but always right truncated. Distributions of (beneficial) fitness effects predicted by a landscape model assuming multiple traits underlying fitness and a single optimum often provide a good description of the empirical distributions in our data. Simulations of data sets containing a mixture of single and double mutants under this landscape show that inferences about the distribution of fitness effects of beneficial mutants is quite robust to contamination by second-site mutations. PMID:21868607

  9. 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.

  10. Chemotactic Motility of Pseudomonas fluorescens F113 under Aerobic and Denitrification Conditions.

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    Candela Muriel

    Full Text Available The sequence of the genome of Pseudomonas fluorescens F113 has shown the presence of multiple traits relevant for rhizosphere colonization and plant growth promotion. Among these traits are denitrification and chemotactic motility. Besides aerobic growth, F113 is able to grow anaerobically using nitrate and nitrite as final electron acceptors. F113 is able to perform swimming motility under aerobic conditions and under anaerobic conditions when nitrate is used as the electron acceptor. However, nitrite can not support swimming motility. Regulation of swimming motility is similar under aerobic and anaerobic conditions, since mutants that are hypermotile under aerobic conditions, such as gacS, sadB, kinB, algU and wspR, are also hypermotile under anaerobic conditions. However, chemotactic behavior is different under aerobic and denitrification conditions. Unlike most pseudomonads, the F113 genome encode three complete chemotaxis systems, Che1, Che2 and Che3. Mutations in each of the cheA genes of the three Che systems has shown that the three systems are functional and independent. Mutation of the cheA1 gene completely abolished swimming motility both under aerobic and denitrification conditions. Mutation of the cheA2 gene, showed only a decrease in swimming motility under both conditions, indicating that this system is not essential for chemotactic motility but is necessary for optimal motility. Mutation of the cheA3 gene abolished motility under denitrification conditions but only produced a decrease in motility under aerobic conditions. The three Che systems proved to be implicated in competitive rhizosphere colonization, being the cheA1 mutant the most affected.

  11. Influencia de la concentración salina y la temperatura en la composición de ácidos grasos de membrana de Pseudomonas fluorescens GNP-OHP-3 Influence of salinity and temperature on fatty acid composition of Pseudomonas fluorescens GNP-OHP-3 membrane

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    G. N. Pucci

    2004-03-01

    Full Text Available Las bacterias responden a los cambios ambientales modificando su composición, para evitar el daño que dichos cambios podrían ejercer. Una de las modificaciones más importantes es la variación de la composición de los ácidos grasos de las membranas celulares, que le permite mantener la homeoviscosidad ante situaciones de estrés. Trabajos previos han estudiado la acción de la temperatura, presión hidrostática y diferentes solventes sobre cepas de Pseudomonas putida. En este trabajo se estudió la acción conjunta de la temperatura y la salinidad sobre la composición de ácidos grasos de membranas celulares de Pseudomonas fluorescens GNP-OHP-3, una cepa bacteriana aislada de un hábitat contaminado con petróleo. Pseudomonas fluorescens GNP-OHP-3 respondió a las variaciones de temperatura modificando los ácidos grasos de sus membranas de manera similar a lo descripto en otros integrantes de su género: ante el aumento de temperatura se observó un incremento de ácidos grasos saturados y una disminución de los ácidos grasos insaturados. En el rango de concentraciones salinas ensayadas las variaciones de los ácidos grasos mayoritarios fueron en general erráticas. La respuesta de los ácidos grasos ciclo propano pudo expresarse con ecuaciones matemáticas que permitieron predecir el porcentaje de estos ácidos en relación a la concentración de cloruro de sodio.The bacteria respond to environmental changes modifying their composition. One of the most important modifications is the variation on fatty acid composition of cellular membranes to maintain the homeoviscosity. The action of temperature, hydrostatic pressure and solvents on Pseudomonas putida has been thoroughly studied. In this paper, the combined action of the temperature and salinity on fatty acid composition of cellular membranes of Pseudomonas fluorescens GNP-OHP-3, a bacterial strain isolated from a petroleum contaminated habitat, was studied. The modifications in the

  12. 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...

  13. Estrategias de identificación de genes de proteasas en una cepa de pseudomonas fluorescens alterante de leche

    OpenAIRE

    Rodrigo Torres, Lidia

    2013-01-01

    La leche es un alimento muy completo nutricionalmente y por ello fácilmente alterable por microorganismos si no se adoptan unas medidas de higiene adecuadas. El análisis de un lote de leche esterilizada comercial alterada reveló la presencia de una cepa de Pseudomonas fluorescens con actividad proteolítica y lipolítica. El objetivo de este trabajo fue diseñar oligonucleótidos y utilizar la técnica de amplificación por PCR (Polymerase Chain Reaction), seguida de secuenciación o de clonación...

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

    Directory of Open Access Journals (Sweden)

    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.

  15. Isolation and characterization of antimicrobial cyclic dipeptides from Pseudomonas fluorescens and their efficacy on sorghum grain mold fungi.

    Science.gov (United States)

    Sajeli Begum, Ahil; Basha, Shaik Ameer; Raghavendra, Govardhanam; Kumar, Mallela Venkata Nagesh; Singh, Yukthi; Patil, Jagannath V; Tanemura, Yuhei; Fujimoto, Yoshinori

    2014-01-01

    This study was aimed at isolation and characterization of natural antifungal compounds for grain mold, a key parasitic fungal disease of sorghum. Pseudomonas fluorescens strain isolated from rhizosphere of groundnut crop was selected as a source. Its biocontrolling ability was assessed by testing some biochemical attributes such as phosphate-solubilization, and HCN, NH3 , indole-3-acetic acid, and siderophore production. The strain showed positive result for all except indole-3-acetic acid, revealing its suitability for a further study. The antibiotic-sensitivity pattern of the strain against 43 antibiotics was also established, which showed resistance to 15 antibiotics. The efficacy of P. fluorescens strain against grain mold was identified by dual culture technique. Hundred percent inhibition was found against Fusarium moniliforme, an important causative agent of this disease. The strain was fermented for secondary metabolites and extracted with AcOEt. Chromatographic separation of the extract yielded four known compounds, cyclo(L-Pro-L-Phe) (1), cyclo(trans-4-hydroxy-L-Pro-L-Leu) (2), cyclo(trans-4-hydroxy-L-Pro-L-Phe) (3), and cyclo(Gly-L-Pro) (4), which were characterized by spectral analysis and optical rotation. The crude extract, a mixture of 2 and 3, and isolated 1 were proved to be significantly effective against grain mold fungi. This is the first report on production of these cyclic dipeptides by P. fluorescens and their antagonistic properties. Copyright © 2014 Verlag Helvetica Chimica Acta AG, Zürich.

  16. Pseudomonas fluorescens induces strain-dependent and strain-independent host plant responses in defense networks, primary metabolism and photosynthesis

    Energy Technology Data Exchange (ETDEWEB)

    Pelletier, Dale A [ORNL; Morrell-Falvey, Jennifer L [ORNL; Karve, Abhijit A [ORNL; Lu, Tse-Yuan S [ORNL; Tschaplinski, Timothy J [ORNL; Tuskan, Gerald A [ORNL; Chen, Jay [ORNL; Martin, Madhavi Z [ORNL; Jawdy, Sara [ORNL; Weston, David [ORNL; Doktycz, Mitchel John [ORNL; Schadt, Christopher Warren [ORNL

    2012-01-01

    Colonization of plants by nonpathogenic Pseudomonas fluorescens strains can confer enhanced defense capacity against a broad spectrum of pathogens. Few studies, however, have linked defense pathway regulation to primary metabolism and physiology. In this study, physiological data, metabolites, and transcript profiles are integrated to elucidate how molecular networks initiated at the root-microbe interface influence shoot metabolism and whole-plant performance. Experiments with Arabidopsis thaliana were performed using the newly identified P. fluorescens GM30 or P. fluorescens Pf-5 strains. Co-expression networks indicated that Pf-5 and GM30 induced a subnetwork specific to roots enriched for genes participating in RNA regulation, protein degradation, and hormonal metabolism. In contrast, only GM30 induced a subnetwork enriched for calcium signaling, sugar and nutrient signaling, and auxin metabolism, suggesting strain dependence in network architecture. In addition, one subnetwork present in shoots was enriched for genes in secondary metabolism, photosynthetic light reactions, and hormone metabolism. Metabolite analysis indicated that this network initiated changes in carbohydrate and amino acid metabolism. Consistent with this, we observed strain-specific responses in tryptophan and phenylalanine abundance. Both strains reduced host plant carbon gain and fitness, yet provided a clear fitness benefit when plants were challenged with the pathogen P. syringae DC3000.

  17. 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.

  18. Listeria monocytogenes Impact on Mature or Old Pseudomonas fluorescens Biofilms During Growth at 4 and 20°C.

    Science.gov (United States)

    Puga, Carmen H; Orgaz, Belen; SanJose, Carmen

    2016-01-01

    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 20°C and 4°C, 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 20°C, but not in those grown at 4°C. 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.

  19. Phytohormone production and colonization of canola (Brassica napus L.) roots by Pseudomonas fluorescens 6-8 under gnotobiotic conditions.

    Science.gov (United States)

    Pallai, Rajash; Hynes, Russell K; Verma, Brij; Nelson, Louise M

    2012-02-01

    Pseudomonas fluorescens 6-8, a rhizosphere isolate previously shown to enhance root elongation of canola ( Brassica napus L.), was characterized for its ability to produce indole-3-acetic acid and cytokinins in pure culture and in the rhizosphere of canola under gnotobiotic conditions in comparison with the cytokinin-producing strain P. fluorescens G20-18 and its mutant CNT2. Strain 6-8 produced isopentenyl adenosine, zeatin riboside, and dihydroxyzeatin riboside at levels similar to those of G20-18, but only very low concentrations of indole-3-acetic acid. In a gnotobiotic assay canola inoculated with 6-8 and G20-18 had higher concentrations of isopentenyl adenosine and zeatin riboside in the rhizosphere and greater root length than the noninoculated control. The ability of strain 6-8 to colonize canola roots was assessed following transformation with the green fluorescent protein and inoculation onto canola seed in a gnotobiotic assay. Higher populations of strain 6-8 were observed on the proximal region of the root closest to the seed than on the mid and distal portions 9 days after seed inoculation. The ability of P. fluorescens 6-8 to produce cytokinins, colonize the roots of canola seedlings, and enhance root elongation may contribute to its ability to survive in the rhizosphere and may benefit seedling growth.

  20. 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.

  1. Cloning and expression of Pseudomonas fluorescens 26-2 lipase gene in Pichia pastoris and characterizing for transesterification.

    Science.gov (United States)

    Yang, Jiangke; Zhang, Bo; Yan, Yunjun

    2009-11-01

    Pseudomonas lipases are important biocatalysts widely used in a variety of industrial fields. An extracellular lipase gene lipA with 1,854-bp open reading frame was cloned from Pseudomonas fluorescens 26-2. The multialignment assay of the putative amino acid and the secondary structure prediction revealed this enzyme could be classified into the lipolytic subfamily I.3 and secreted via adenosine-triphosphate-binding cassette pathway. The lipA gene was integrated into Pichia pastoris GS115, and the methanol-inducible recombinants with Mut(S) and Mut(+) phenotypes were acquired. The characteristics and the transesterification capacity shown by this enzyme suggested it is a useful biocatalyst for biodiesel preparation.

  2. Identification of anthranilate and benzoate metabolic operons of Pseudomonas fluorescens and functional characterization of their promoter regions

    Directory of Open Access Journals (Sweden)

    Lee Vincent D

    2006-01-01

    Full Text Available Abstract Background In an effort to identify alternate recombinant gene expression systems in Pseudomonas fluorescens, we identified genes encoding two native metabolic pathways that were inducible with inexpensive compounds: the anthranilate operon (antABC and the benzoate operon (benABCD. Results The antABC and benABCD operons were identified by homology to the Acinetobacter sp. anthranilate operon and Pseudomonas putida benzoate operon, and were confirmed to be regulated by anthranilate or benzoate, respectively. Fusions of the putative promoter regions to the E. coli lacZ gene were constructed to confirm inducible gene expression. Each operon was found to be controlled by an AraC family transcriptional activator, located immediately upstream of the first structural gene in each respective operon (antR or benR. Conclusion We have found the anthranilate and benzoate promoters to be useful for tightly controlling recombinant gene expression at both small (

  3. Mutation rate, spectrum, topology, and context-dependency in the DNA mismatch repair-deficient Pseudomonas fluorescens ATCC948.

    Science.gov (United States)

    Long, Hongan; Sung, Way; Miller, Samuel F; Ackerman, Matthew S; Doak, Thomas G; Lynch, Michael

    2014-12-23

    High levels of genetic diversity exist among natural isolates of the bacterium Pseudomonas fluorescens, and are especially elevated around the replication terminus of the genome, where strain-specific genes are found. In an effort to understand the role of genetic variation in the evolution of Pseudomonas, we analyzed 31,106 base substitutions from 45 mutation accumulation lines of P. fluorescens ATCC948, naturally deficient for mismatch repair, yielding a base-substitution mutation rate of 2.34 × 10(-8) per site per generation (SE: 0.01 × 10(-8)) and a small-insertion-deletion mutation rate of 1.65 × 10(-9) per site per generation (SE: 0.03 × 10(-9)). We find that the spectrum of mutations in prophage regions, which often contain virulence factors and antibiotic resistance, is highly similar to that in the intergenic regions of the host genome. Our results show that the mutation rate varies around the chromosome, with the lowest mutation rate found near the origin of replication. Consistent with observations from other studies, we find that site-specific mutation rates are heavily influenced by the immediately flanking nucleotides, indicating that mutations are context dependent. © The Author(s) 2014. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

  4. 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.

  5. Synthesis of silver nanoparticles by endosymbiont Pseudomonas fluorescens CA 417 and their bactericidal activity.

    Science.gov (United States)

    Syed, Baker; M N, Nagendra Prasad; B L, Dhananjaya; K, Mohan Kumar; S, Yallappa; S, Satish

    2016-12-01

    The present study emphasizes on biogenic synthesis of silver nanoparticles and their bactericidal activity against human and phytopathogens. Nanoparticle synthesis was performed using endosymbiont Pseudomonas fluorescens CA 417 inhabiting Coffea arabica L. Synthesized nanoparticles were characterized using hyphenated spectroscopic techniques such as UV-vis spectroscopy which revealed maximum absorption 425nm. Fourier transform infrared spectroscopy (FTIR) analysis revealed the possible functional groups mediating and stabilizing silver nanoparticles with predominant peaks occurring at 3346 corresponding to hydroxyl group, 1635 corresponding carbonyl group and 680 to aromatic group. X-ray diffraction (XRD) analysis revealed the Bragg's diffraction pattern with distinct peaks at 38° 44°, 64° and 78° revealing the face-centered cubic (fcc) metallic crystal corresponding to the (111), (200), (220) and (311) facets of the crystal planes at 2θ angle. The energy dispersive X-ray spectroscopy (EDS) analysis revealed presence of high intense absorption peak at 3keV is a typical characteristic of nano-crystalline silver which confirmed the presence of elemental silver. TEM analysis revealed the size of the nanoparticles to be in the range 5-50nm with polydisperse nature of synthesized nanoparticles bearing myriad shapes. The particle size determined by Dynamic light scattering (DLS) method revealed average size to be 20.66nm. The synthesized silver nanoparticles exhibited significant antibacterial activity against panel of test pathogens. The results showed Klebsiella pneumoniae (MTCC 7407) and Xanthomonas campestris to be more sensitive among the test human pathogen and phyto-pathogen respectively. The study also reports synergistic effect of silver nanoparticles in combination with kanamycin which displayed increased fold activity up to 58.3% against Klebsiella pneumoniae (MTCC 7407). The results of the present investigation are promising enough and attribute towards

  6. 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

    An antibiotic substance isolated from Pseudomonas fluorescens strain G308 was earlier assigned the structure of N-mercapto-4-formylcarbostyril, but computational predictions of the H-1 and C-13 NMR magnetic shielding tensors show this structure to be incompatible with the published spectroscopic...

  7. Expression of Fap amyloids in Pseudomonas aeruginosa, P. fluorescens, and P. putida results in aggregation and increased biofilm formation

    DEFF Research Database (Denmark)

    Dueholm, Morten S; Søndergaard, Mads; Nilsson, Martin

    2013-01-01

    The fap operon, encoding functional amyloids in Pseudomonas (Fap), is present in most pseudomonads, but so far the expression and importance for biofilm formation has only been investigated for P. fluorescens strain UK4. In this study, we demonstrate the capacity of P. aeruginosa PAO1, P. fluores...

  8. 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

    The antagonistic interaction between a potential fish probiont, Pseudomonas fluorescens strain AH2, and its target organism, Vibrio anguillarum, was investigated by studying the genetic response of the target organism when it was exposed to the antagonist. We compared the differential display of ......, which results in instant iron deprivation of the pathogen V. anguillarum and complete growth arrest....

  9. The Rsm regulon of plant growth-promoting Pseudomonas fluorescens SS101: role of small RNAs in regulation of lipopeptide biosynthesis

    NARCIS (Netherlands)

    Song, C.; Voort, van der M.; Mortel, van de J.; Hassan, K.A.; Elbourne, L.D.H.; Paulsen, I.T.; Loper, J.E.; Raaijmakers, J.M.

    2015-01-01

    The rhizobacterium Pseudomonas fluorescens SS101 inhibits growth of oomycete and fungal pathogens, and induces resistance in plants against pathogens and insects. To unravel regulatory pathways of secondary metabolite production in SS101, we conducted a genome-wide search for sRNAs and performed

  10. New insights into Pseudomonas fluorescens alginate biosynthesis relevant for the establishment of an efficient production process for microbial alginates.

    Science.gov (United States)

    Maleki, Susan; Mærk, Mali; Hrudikova, Radka; Valla, Svein; Ertesvåg, Helga

    2017-07-25

    Alginate denotes a family of linear polysaccharides with a wide range of industrial and pharmaceutical applications. Presently, all commercially available alginates are manufactured from brown algae. However, bacterial alginates have advantages with regard to compositional homogeneity and reproducibility. In order to be able to design bacterial strains that are better suited for industrial alginate production, defining limiting factors for alginate biosynthesis is of vital importance. Our group has been studying alginate biosynthesis in Pseudomonas fluorescens using several complementary approaches. Alginate is synthesised and transported out of the cell by a multiprotein complex spanning from the inner to the outer membrane. We have developed an immunogold labelling procedure in which the porin AlgE, as a part of this alginate factory, could be detected by transmission electron microscopy. No time-dependent correlation between the number of such factories on the cell surface and alginate production level was found in alginate-producing strains. Alginate biosynthesis competes with the central carbon metabolism for the key metabolite fructose 6-phosphate. In P. fluorescens, glucose, fructose and glycerol, are metabolised via the Entner-Doudoroff and pentose phosphate pathways. Mutational analysis revealed that disruption of the glucose 6-phosphate dehydrogenase gene zwf-1 resulted in increased alginate production when glycerol was used as carbon source. Furthermore, alginate-producing P. fluorescens strains cultivated on glucose experience acid stress due to the simultaneous production of alginate and gluconate. The combined results from our studies strongly indicate that the availability of fructose 6-phosphate and energy requires more attention in further research aimed at the development of an optimised alginate production process. Copyright © 2016 Elsevier B.V. All rights reserved.

  11. Susceptibility of Mycobacterium immunogenum and Pseudomonas fluorescens to formaldehyde and non-formaldehyde biocides in semi-synthetic metalworking fluids.

    Science.gov (United States)

    Selvaraju, Suresh B; Khan, Izhar U H; Yadav, Jagjit S

    2011-01-20

    Mycobacterium immunogenum, a newly identified member of the Mycobacterium chelonae_M. abscessus complex is considered a potential etiological agent for hypersensitivity pneumonitis (HP) in machine workers exposed to contaminated metalworking fluid (MWF). This study investigated the biocidal efficacy of the frequently applied commercial formaldehyde-releasing (HCHO) biocides Grotan and Bioban CS 1135 and non-HCHO type biocides Kathon 886 MW (isothiazolone) and Preventol CMK 40 (phenolic) toward this emerging mycobacterial species (M. immunogenum) in HP-linked MWFs, alone and in presence of a representative of the Gram-negative bacterial contaminants, Pseudomonas fluorescens, using two semi-synthetic MWF matrices (designated Fluid A and Fluid B). Relative biocide susceptibility analysis indicated M immunogenum to be comparatively more resistant (2-1600 fold) than P. fluorescens to the tested biocides under the varied test conditions. In terms of minimum inhibitory concentration, Kathon was the most effective biocide against M. immunogenum. Fluid factors had a major effect on the biocide susceptibility. Fluid A formulation provided greater protective advantage to the test organisms than Fluid B. Fluid dialysis (Fluid A) led to an increased biocidal efficacy of Grotan, Kathon and Preventol against M. immunogenum further implying the role of native fluid components. Used fluid matrix, in general, increased the resistance of the two test organisms against the biocides, with certain exceptions. M. immunogenum resistance increased in presence of the co-contaminant P. fluorescens. Collectively, the results show a multifactorial nature of the biocide susceptibility of MWF-colonizing mycobacteria and highlight the importance of more rigorous efficacy testing and validation of biocides prior to and during their application in metalworking fluid operations.

  12. Susceptibility of Mycobacterium immunogenum and Pseudomonas fluorescens to Formaldehyde and Non-Formaldehyde Biocides in Semi-Synthetic Metalworking Fluids

    Directory of Open Access Journals (Sweden)

    Suresh B. Selvaraju

    2011-01-01

    Full Text Available Mycobacterium immunogenum, a newly identified member of the Mycobacterium chelonae_M. abscessus complex is considered a potential etiological agent for hypersensitivity pneumonitis (HP in machine workers exposed to contaminated metalworking fluid (MWF. This study investigated the biocidal efficacy of the frequently applied commercial formaldehyde-releasing (HCHO biocides Grotan and Bioban CS 1135 and non-HCHO type biocides Kathon 886 MW (isothiazolone and Preventol CMK 40 (phenolic toward this emerging mycobacterial species (M. immunogenum in HP-linked MWFs, alone and in presence of a representative of the Gram-negative bacterial contaminants, Pseudomonas fluorescens, using two semi-synthetic MWF matrices (designated Fluid A and Fluid B. Relative biocide susceptibility analysis indicated M immunogenum to be comparatively more resistant (2–1600 fold than P. fluorescens to the tested biocides under the varied test conditions. In terms of minimum inhibitory concentration, Kathon was the most effective biocide against M. immunogenum. Fluid factors had a major effect on the biocide susceptibility. Fluid A formulation provided greater protective advantage to the test organisms than Fluid B. Fluid dialysis (Fluid A led to an increased biocidal efficacy of Grotan, Kathon and Preventol against M. immunogenum further implying the role of native fluid components. Used fluid matrix, in general, increased the resistance of the two test organisms against the biocides, with certain exceptions. M. immunogenum resistance increased in presence of the co-contaminant P. fluorescens. Collectively, the results show a multifactorial nature of the biocide susceptibility of MWF-colonizing mycobacteria and highlight the importance of more rigorous efficacy testing and validation of biocides prior to and during their application in metalworking fluid operations.

  13. Enhanced citric acid biosynthesis in Pseudomonas fluorescens ATCC 13525 by overexpression of the Escherichia coli citrate synthase gene.

    Science.gov (United States)

    Buch, Aditi D; Archana, G; Kumar, G Naresh

    2009-08-01

    Citric acid secretion by fluorescent pseudomonads has a distinct significance in microbial phosphate solubilization. The role of citrate synthase in citric acid biosynthesis and glucose catabolism in pseudomonads was investigated by overexpressing the Escherichia coli citrate synthase (gltA) gene in Pseudomonas fluorescens ATCC 13525. The resultant approximately 2-fold increase in citrate synthase activity in the gltA-overexpressing strain Pf(pAB7) enhanced the intracellular and extracellular citric acid yields during the stationary phase, by about 2- and 26-fold, respectively, as compared to the control, without affecting the growth rate, glucose depletion rate or biomass yield. Decreased glucose consumption was paralleled by increased gluconic acid production due to an increase in glucose dehydrogenase activity. While the extracellular acetic acid yield increased in Pf(pAB7), pyruvic acid secretion decreased, correlating with an increase in pyruvate carboxylase activity and suggesting an increased demand for the anabolic precursor oxaloacetate. Activities of two other key enzymes, glucose-6-phosphate dehydrogenase and isocitrate dehydrogenase, remained unaltered, and the contribution of phosphoenolpyruvate carboxylase and isocitrate lyase to glucose catabolism was negligible. Strain Pf(pAB7) demonstrated an enhanced phosphate-solubilizing ability compared to the control. Co-expression of the Synechococcus elongatus PCC 6301 phosphoenolpyruvate carboxylase and E. coli gltA genes in P. fluorescens ATCC 13525, so as to supplement oxaloacetate for citrate biosynthesis, neither significantly affected citrate biosynthesis nor caused any change in the other physiological and biochemical parameters measured, despite approximately 1.3- and 5-fold increases in citrate synthase and phosphoenolpyruvate carboxylase activities, respectively. Thus, our results demonstrate that citrate synthase is rate-limiting in enhancing citrate biosynthesis in P. fluorescens ATCC 13525

  14. Influence of Arbuscular Mycorrhizal Fungi and Pseudomonas fluorescens at Different Superphosphate Levels on Linseed (Linum usitatissimum L. Growth Response Influencia de Hongos Micorriza Arbusculares y Pseudomonas fluorescens con Diferentes Niveles de Superfosfato sobre la Respuesta al Crecimiento de Lino (Linum usitatissimum L.

    Directory of Open Access Journals (Sweden)

    Neetu Neetu

    2012-06-01

    Full Text Available The aim of this study was to investigate the influence of arbuscular mycorrhizal fungi (AMF Glomus mosseae (T.H. Nicolson & Gerd. Gerd. & Trappe and Acaulospora laevis (Gerd. & Trappe on linseed (Linum usitatissimum L. growth response with phosphate solubilizing bacteria Pseudomonas fluorescens; different doses of superphosphate were used: 20 kg ha4 (half recommended dose, 40 kg ha4 (recommended dose, and 80 kg ha4 (double recommended dose in earthen pots filled with sterilized soil under greenhouse conditions. Among all the growth parameters, the following were the highest in the G. mosseae + P. fluorescens combination at the medium concentration (recommended superphosphate dose: plant height (78.74 ± 1.8 cm, fresh shoot weight (3.45 ± 0.294 g, dry shoot weight (0.57 ± 0.007 g, fresh root weight (0.223 ± 0.023 g, dry root weight (0.036 ± 0.004 g, root length (17.67 ± 0.48 cm, AM spore number (94.4 ± 9.86, shoot (1.14 ± 0.115% and root (1.29 ± 0.110% P content, and acidic (0.447 ± 0.012 IU g-1 FW and alkaline phosphatase activity (0.119 ± 0.008 IU g-1 FW. The percentage mycorrhizal root colonization with the A. laevis + P. fluorescens (86.86 ± 2.17% combination and chlorophyll content with the G. mosseae + A. laevis + P. fluorescens (0.474 ± 0.009 mg g-1 FW combination recorded the highest values at the low concentration (half recommended superphosphate dose as compared with non-mycorrhizal plants (control. The high superphosphate dose clearly reduced or decreased all the growth parameters. Therefore, vigorous growth and maximum flax yield can be achieved by inoculating plants with AM fungi and P. fluorescens with the recommended dose or less than the recommended dose of superphosphate.La presente investigation tuvo como objetivo estudiar la influencia de hongos micorrícicos arbusculares (AMF, i.e., Glomus mosseae (T.H. Nicolson & Gerd. Gerd. & Trappey Acaulospora laevis (Gerd. & Trappe, en la respuesta de crecimiento de lino

  15. The use of Pseudomonas fluorescens P13 to control sclerotinia stem rot (Sclerotinia sclerotiorum) of oilseed rape.

    Science.gov (United States)

    Li, Hui; Li, Huaibo; Bai, Yan; Wang, Jing; Nie, Ming; Li, Bo; Xiao, Ming

    2011-12-01

    Sclerotinia stem rot (SSR) caused by the fungus Sclerotinia sclerotiorum has been an increasing threat to oilseed rape (Brassica napus L.) cultivation. Efficient and environment-friendly treatments are much needed. Here we focus on microbial control. The Pseudomonas fluorescens P13 that was isolated from oilseed rape cultivation soil, proved to be a useful biocontrol strain for application. Morphology, physiological and biochemical tests and 16S rDNA analysis demonstrated that it was P. fluorescens P13 and that it had a broad antagonistic spectrum, significantly lessening the mycelial growth of S. sclerotiorum by 84.4% and suppressing sclerotial formation by 95-100%. Scanning electron microscopy studies attested that P13 deformed S. sclerotiorum mycelia when they were cultured together. P13 did not produce chitinase but did produce hydrogen cyanide (HCN) which was likely one of the antagonistic mechanisms. The density of P13 remained at a high level (≥10(6) CFU/ml) during 5 weeks in the rhizosphere soil and roots. P13 reduced SSR severity at least by 59% in field studies and also promoted seedling growth (psclerotiorum.

  16. 1-aminocyclopropane-1-carboxylate deaminase from Pseudomonas fluorescens promoting the growth of Chinese cabbage and its polyclonal antibody.

    Science.gov (United States)

    Soh, Byoung Yul; Lee, Gun Woong; Go, Eun Byeul; Kim, Byeo-Ri; Lee, Kui-Jae; Chae, Jong-Chan

    2014-05-01

    Bacterial 1-aminocyclopropane-1-carboxlyate (ACC) deaminase (AcdS) is an enzyme that cleaves ACC, a precursor of the plant hormone ethylene, into α-ketobutyrate and ammonia. The acdS gene was cloned from Pseudomonas fluorescens, which was capable of improving the seedling of Chinese cabbage under salinity condition. The recombinant AcdS (rAcdS) exhibited optimal activity at pH 8.5 and 30°C. Strong activity was sustained at up to 100 mM NaCl. The polyclonal anti-P. fluorescens AcdS antibody was produced in a rabbit that had been immunized with the purified rAcdS. This antibody successfully recognized the homologous antigens derived from the total proteins of isolated plant growth-promoting microorganisms. A statistically significant correlation was observed between the intensity of hybridization signal and AcdS activity measured by a biochemical method, suggesting its application as a useful indicator for active deaminases.

  17. 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]).

  18. Influence of phenolic acids on indole acetic acid production and on the type III secretion system gene transcription in food-associated Pseudomonas fluorescens KM05.

    Science.gov (United States)

    Myszka, Kamila; Schmidt, Marcin T; Olejnik-Schmidt, Agnieszka K; Leja, Katarzyna; Czaczyk, Katarzyna

    2014-12-01

    The purpose of these investigations was to evaluate the reduction capability of phenolic acids (ferulic, chlorogenic, gallic, and p-coumaric acids) on indole acetic acid synthesis by food-associated Pseudomonas fluorescens KM05. Specific genetic primer for the type III secretion system (TTSS) in P. fluorescens KM05 was designed and the influence of phenolic acids on its expression was investigated. In the work the ferulic and chlorogenic acids at the concentration of 0.02 and 0.04 μg/ml affected on bacterial growth pattern and the signal molecules production. The phenolic acids, that were appreciable effective against P. fluorescens KM05 indole acetic acid production, significantly suppressed TTSS gene. Copyright © 2014 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

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

    National Research Council Canada - National Science Library

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

    2013-01-01

    .... 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...

  20. Proteomic Detection of Non-Annotated Protein-Coding Genes in Pseudomonas fluorescens Pf0-1

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Wook; Silby, Mark W.; Purvine, Samuel O.; Nicoll, Julie S.; Hixson, Kim K.; Monroe, Matthew E.; Nicora, Carrie D.; Lipton, Mary S.; Levy, Stuart B.

    2009-12-24

    Genome sequences are annotated by computational prediction of coding sequences, followed by similarity searches such as BLAST, which provide a layer of (possible) functional information. While the existence of processes such as alternative splicing complicates matters for eukaryote genomes, the view of bacterial genomes as a linear series of closely spaced genes leads to the assumption that computational annotations which predict such arrangements completely describe the coding capacity of bacterial genomes. We undertook a proteomic study to identify proteins expressed by Pseudomonas fluorescens Pf0-1 from genes which were not predicted during the genome annotation. Mapping peptides to the Pf0-1 genome sequence identified sixteen non-annotated protein-coding regions, of which nine were antisense to predicted genes, six were intergenic, and one read in the same direction as an annotated gene but in a different frame. The expression of all but one of the newly discovered genes was verified by RT-PCR. Few clues as to the function of the new genes were gleaned from informatic analyses, but potential orthologues in other Pseudomonas genomes were identified for eight of the new genes. The 16 newly identified genes improve the quality of the Pf0-1 genome annotation, and the detection of antisense protein-coding genes indicates the under-appreciated complexity of bacterial genome organization.

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

    Directory of Open Access Journals (Sweden)

    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. 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

    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-p...... that environmental elicitors may also influence nunF expression which upon activation regulates nunamycin and nunapeptin production required for the growth inhibition of phytopathogens....

  3. Stabilitäten der Benzaldehydlyase aus Pseudomonas fluorescens und der Carbonylreduktase aus Candida parapsilosis in wässrig-organischen Zweiphasensystemen

    OpenAIRE

    Van Den Wittenboer, Anne

    2009-01-01

    Benzaldehyde lyase from Pseudomonas fluorescens (BAL) and carbonylreductase from Candida parapsilosis (CPCR) are versatile catalysts for the production of various chiral intermediates, which are of interest for the pharmaceutical industry. Due to the mostly hydrophobic reactants only low productivities can be achieved in aqueous reaction systems and thus the use of alternative reaction media is desirable. Especially the use of aqueous-organic biphasic systems is promising, since the extractio...

  4. A modular esterase from Pseudomonas fluorescens subsp. cellulosa contains a non-catalytic cellulose-binding domain.

    Science.gov (United States)

    Ferreira, L M; Wood, T M; Williamson, G; Faulds, C; Hazlewood, G P; Black, G W; Gilbert, H J

    1993-09-01

    The 5' regions of genes xynB and xynC, coding for a xylanase and arabinofuranosidase respectively, are identical and are reiterated four times within the Pseudomonas fluorescens subsp. cellulosa genome. To isolate further copies of the reiterated xynB/C 5' region, a genomic library of Ps. fluorescens subsp. cellulosa DNA was screened with a probe constructed from the conserved region of xynB. DNA from one phage which hybridized to the probe, but not to sequences upstream or downstream of the reiterated xynB/C locus, was subcloned into pMTL22p to construct pFG1. The recombinant plasmid expressed a protein in Escherichia coli, designated esterase XYLD, of M(r) 58,500 which bound to cellulose but not to xylan. XYLD hydrolysed aryl esters, released acetate groups from acetylxylan and liberated 4-hydroxy-3-methoxycinnamic acid from destarched wheat bran. The nucleotide sequence of the XYLD-encoding gene, xynD, revealed an open reading frame of 1752 bp which directed the synthesis of a protein of M(r) 60,589. The 5' 817 bp of xynD and the amino acid sequence between residues 37 and 311 of XYLD were almost identical with the corresponding regions of xynB and xynC and their encoded proteins XYLB and XYLC. Truncated derivatives of XYLD lacking the N-terminal conserved sequence retained the capacity to hydrolyse ester linkages, but did not bind cellulose. Expression of truncated derivatives of xynD, comprising the 5' 817 bp sequence, encoded a non-catalytic polypeptide that bound cellulose. These data indicate that XYLD has a modular structure comprising of a N-terminal cellulose-binding domain and a C-terminal catalytic domain.

  5. Biological control of wheat root diseases by the CLP-producing strain Pseudomonas fluorescens HC1-07.

    Science.gov (United States)

    Yang, Ming-Ming; Wen, Shan-Shan; Mavrodi, Dmitri V; Mavrodi, Olga V; von Wettstein, Diter; Thomashow, Linda S; Guo, Jian-Hua; Weller, David M

    2014-03-01

    Pseudomonas fluorescens HC1-07, previously isolated from the phyllosphere of wheat grown in Hebei province, China, suppresses the soilborne disease of wheat take-all, caused by Gaeumannomyces graminis var. tritici. We report here that strain HC1-07 also suppresses Rhizoctonia root rot of wheat caused by Rhizoctonia solani AG-8. Strain HC1-07 produced a cyclic lipopeptide (CLP) with a molecular weight of 1,126.42 based on analysis by electrospray ionization mass spectrometry. Extracted CLP inhibited the growth of G. graminis var. tritici and R. solani in vitro. To determine the role of this CLP in biological control, plasposon mutagenesis was used to generate two nonproducing mutants, HC1-07viscB and HC1-07prtR2. Analysis of regions flanking plasposon insertions in HC1-07prtR2 and HC1-07viscB revealed that the inactivated genes were similar to prtR and viscB, respectively, of the well-described biocontrol strain P. fluorescens SBW25 that produces the CLP viscosin. Both genes in HC1-07 were required for the production of the viscosin-like CLP. The two mutants were less inhibitory to G. graminis var. tritici and R. solani in vitro and reduced in ability to suppress take-all. HC1-07viscB but not HC-07prtR2 was reduced in ability to suppress Rhizoctonia root rot. In addition to CLP production, prtR also played a role in protease production.

  6. 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.

  7. Functional Characterization of the Mannitol Promoter of Pseudomonas fluorescens DSM 50106 and Its Application for a Mannitol-Inducible Expression System for Pseudomonas putida KT2440.

    Directory of Open Access Journals (Sweden)

    Jana Hoffmann

    Full Text Available A new pBBR1MCS-2-derived vector containing the Pseudomonas fluorescens DSM10506 mannitol promoter PmtlE and mtlR encoding its AraC/XylS type transcriptional activator was constructed and optimized for low basal expression. Mannitol, arabitol, and glucitol-inducible gene expression was demonstrated with Pseudomonas putida and eGFP as reporter gene. The new vector was applied for functional characterization of PmtlE. Identification of the DNA binding site of MtlR was achieved by in vivo eGFP measurement with PmtlE wild type and mutants thereof. Moreover, purified MtlR was applied for detailed in vitro investigations using electrophoretic mobility shift assays and DNaseI footprinting experiments. The obtained data suggest that MtlR binds to PmtlE as a dimer. The proposed DNA binding site of MtlR is AGTGC-N5-AGTAT-N7-AGTGC-N5-AGGAT. The transcription activation mechanism includes two binding sites with different binding affinities, a strong upstream binding site and a weaker downstream binding site. The presence of the weak downstream binding site was shown to be necessary to sustain mannitol-inducibility of PmtlE. Two possible functions of mannitol are discussed; the effector might stabilize binding of the second monomer to the downstream half site or promote transcription activation by inducing a conformational change of the regulator that influences the contact to the RNA polymerase.

  8. Functional Characterization of the Mannitol Promoter of Pseudomonas fluorescens DSM 50106 and Its Application for a Mannitol-Inducible Expression System for Pseudomonas putida KT2440

    Science.gov (United States)

    Hoffmann, Jana; Altenbuchner, Josef

    2015-01-01

    A new pBBR1MCS-2-derived vector containing the Pseudomonas fluorescens DSM10506 mannitol promoter PmtlE and mtlR encoding its AraC/XylS type transcriptional activator was constructed and optimized for low basal expression. Mannitol, arabitol, and glucitol-inducible gene expression was demonstrated with Pseudomonas putida and eGFP as reporter gene. The new vector was applied for functional characterization of PmtlE. Identification of the DNA binding site of MtlR was achieved by in vivo eGFP measurement with PmtlE wild type and mutants thereof. Moreover, purified MtlR was applied for detailed in vitro investigations using electrophoretic mobility shift assays and DNaseI footprinting experiments. The obtained data suggest that MtlR binds to PmtlE as a dimer. The proposed DNA binding site of MtlR is AGTGC-N5-AGTAT-N7-AGTGC-N5-AGGAT. The transcription activation mechanism includes two binding sites with different binding affinities, a strong upstream binding site and a weaker downstream binding site. The presence of the weak downstream binding site was shown to be necessary to sustain mannitol-inducibility of PmtlE. Two possible functions of mannitol are discussed; the effector might stabilize binding of the second monomer to the downstream half site or promote transcription activation by inducing a conformational change of the regulator that influences the contact to the RNA polymerase. PMID:26207762

  9. In vitro effects of Laccaria bicolor S238 N and Pseudomonas fluorescens strain BBc6 on rooting of de-rooted shoot hypocotyls of Norway spruce.

    Science.gov (United States)

    Karabaghli, C.; Frey-Klett, P.; Sotta, B.; Bonnet, M.; Le Tacon, F.

    1998-02-01

    The ectomycorrhizal fungus Laccaria bicolor S238 N and the bacterium Pseudomonas fluorescens BBc6 were used separately and in combination to induce in vitro rooting of de-rooted shoot hypocotyls of Norway spruce (Picea abies (L.) Karst.). When the culture medium was supplemented with tryptophan, a precursor of indole-3-acetic acid (IAA) synthesis, the presence of the ectomycorrhizal fungus increased the percentage of hypocotyls forming roots; furthermore, both the fungal and bacterial inoculations enhanced the number of roots formed per rooted hypocotyl. Similar results were obtained by adding exogenous IAA (5 and 10 &mgr;M) to the rooting medium. After the rooting phase, the fungal inoculation enhanced adventitious root elongation and branching as well as the aerial growth of the cuttings. Pseudomonas fluorescens BBc6 had no effect on these parameters. The production of IAA by pure cultures of L. bicolor S238 N and P. fluorescens BBc6 was estimated by immunochemical analysis using specific anti-IAA antibodies. Both L. bicolor S238 N and P. fluorescens BBc6 synthesized IAA in pure culture and synthesis was stimulated in the presence of tryptophan. Thus, the effect of the fungus in stimulating adventitious root formation and subsequent elongation and branching can be attributed, at least partially, to the synthesis of IAA by the fungus. The finding that P. fluorescens BBc6 had no effect on root elongation and branching although it produced IAA suggests that either IAA was not the only parameter involved in the stimulation of these processes by L. bicolor S238 N or the bacterium produced other compounds that counteracted the stimulatory effects of IAA on root elongation and branching.

  10. 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.

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

    DEFF Research Database (Denmark)

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

    sequencing was conducted to investigate the response of the bacterium to different plant pathogens in dual-culture. Results/Discussion: Key biocontrol components of P. fluorescens In5 identified to date include the two secondary metabolites nunamycin and nunapeptin that form part of a complex interaction...... for combatting plant pathogenic fungi. Current studies are aimed at unravelling further the complex mode of action underpinning the antagonistic activity of In5 in order to develop effective microbial biocontrol agents (mBCAs) for the management of soil-borne plant diseases.......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...

  12. Effect of biocontrol agent Pseudomonas fluorescens 2P24 on soil fungal community in cucumber rhizosphere using T-RFLP and DGGE.

    Science.gov (United States)

    Gao, Guanpeng; Yin, Danhan; Chen, Shengju; Xia, Fei; Yang, Jie; Li, Qing; Wang, Wei

    2012-01-01

    Fungi and fungal community play important roles in the soil ecosystem, and the diversity of fungal community could act as natural antagonists of various plant pathogens. Biological control is a promising method to protect plants as chemical pesticides may cause environment pollution. Pseudomonas fluorescens 2P24 had strong inhibitory on Rastonia solanacearum, Fusarium oxysporum and Rhizoctonia solani, etc., and was isolated from the wheat rhizosphere take-all decline soils in Shandong province, China. However, its potential effect on soil fungal community was still unknown. In this study, the gfp-labeled P. fluorescens 2P24 was inoculated into cucumber rhizosphere, and the survival of 2P24 was monitored weekly. The amount decreased from 10(8) to 10(5) CFU/g dry soils. The effect of 2P24 on soil fungal community in cucumber rhizosphere was investigated using T-RFLP and DGGE. In T-RFLP analysis, principle component analysis showed that the soil fungal community was greatly influenced at first, digested with restriction enzyme Hinf I and Taq I. However, there was little difference as digested by different enzymes. DGGE results demonstrated that the soil fungal community was greatly shocked at the beginning, but it recovered slowly with the decline of P. fluorescens 2P24. Four weeks later, there was little difference between the treatment and control. Generally speaking, the effect of P. fluorescens 2P24 on soil fungal community in cucumber rhizosphere was just transient.

  13. Effect of biocontrol agent Pseudomonas fluorescens 2P24 on soil fungal community in cucumber rhizosphere using T-RFLP and DGGE.

    Directory of Open Access Journals (Sweden)

    Guanpeng Gao

    Full Text Available Fungi and fungal community play important roles in the soil ecosystem, and the diversity of fungal community could act as natural antagonists of various plant pathogens. Biological control is a promising method to protect plants as chemical pesticides may cause environment pollution. Pseudomonas fluorescens 2P24 had strong inhibitory on Rastonia solanacearum, Fusarium oxysporum and Rhizoctonia solani, etc., and was isolated from the wheat rhizosphere take-all decline soils in Shandong province, China. However, its potential effect on soil fungal community was still unknown. In this study, the gfp-labeled P. fluorescens 2P24 was inoculated into cucumber rhizosphere, and the survival of 2P24 was monitored weekly. The amount decreased from 10(8 to 10(5 CFU/g dry soils. The effect of 2P24 on soil fungal community in cucumber rhizosphere was investigated using T-RFLP and DGGE. In T-RFLP analysis, principle component analysis showed that the soil fungal community was greatly influenced at first, digested with restriction enzyme Hinf I and Taq I. However, there was little difference as digested by different enzymes. DGGE results demonstrated that the soil fungal community was greatly shocked at the beginning, but it recovered slowly with the decline of P. fluorescens 2P24. Four weeks later, there was little difference between the treatment and control. Generally speaking, the effect of P. fluorescens 2P24 on soil fungal community in cucumber rhizosphere was just transient.

  14. Virulence of the Pseudomonas fluorescens clinical strain MFN1032 towards Dictyostelium discoideum and macrophages in relation with type III secretion system

    Directory of Open Access Journals (Sweden)

    Sperandio Daniel

    2012-09-01

    Full Text Available Abstract Background Pseudomonas fluorescens biovar I MFN1032 is a clinical isolate able to grow at 37°C. This strain displays secretion-mediated hemolytic activity involving phospholipase C and cyclolipopeptides, and a cell-associated hemolytic activity distinct from the secreted hemolytic activity. Cell-associated hemolysis is independent of biosurfactant production and remains in a gacA mutant. Disruption of the hrpU-like operon (the basal part of type III secretion system from rhizospheric strains suppresses this activity. We hypothesized that this phenotype could reflect evolution of an ancestral mechanism involved in the survival of this species in its natural niche. In this study, we evaluated the hrpU-like operon’s contribution to other virulence mechanisms using a panel of Pseudomonas strains from various sources. Results We found that MFN1032 inhibited the growth of the amoebae Dictyostelium discoideum and that this inhibition involved the hrpU-like operon and was absent in a gacA mutant. MFN1032 was capable of causing macrophage lysis, if the hrpU-like operon was intact, and this cytotoxicity remained in a gacA mutant. Cell-associated hemolytic activity and macrophage necrosis were found in other P. fluorescens clinical isolates, but not in biocontrol P. fluorescens strains harbouring hrpU-like operon. The growth of Dictyostelium discoideum was inhibited to a different extent by P. fluorescens strains without correlation between this inhibition and hrpU-like operon sequences. Conclusions In P. fluorescens MFN1032, the basal part of type III secretion system plays a role in D. discoideum growth inhibition and macrophage necrosis. The inhibition of D. discoideum growth is dependent on the GacS/GacA system, while cell-associated hemolytic activity and macrophage lysis are not. Virulence against eukaryotic cells based on the hrpU-like operon may be more than just a stochastic evolution of a conserved system dedicated to survival in

  15. Metabolic and Transcriptomic Changes Induced in Arabidopsis by the Rhizobacterium Pseudomonas fluorescens SS1011[W][OA

    Science.gov (United States)

    van de Mortel, Judith E.; de Vos, Ric C.H.; Dekkers, Ester; Pineda, Ana; Guillod, Leandre; Bouwmeester, Klaas; van Loon, Joop J.A.; Dicke, Marcel; Raaijmakers, Jos M.

    2012-01-01

    Systemic resistance induced in plants by nonpathogenic rhizobacteria is typically effective against multiple pathogens. Here, we show that root-colonizing Pseudomonas fluorescens strain SS101 (Pf.SS101) enhanced resistance in Arabidopsis (Arabidopsis thaliana) against several bacterial pathogens, including Pseudomonas syringae pv tomato (Pst) and the insect pest Spodoptera exigua. Transcriptomic analysis and bioassays with specific Arabidopsis mutants revealed that, unlike many other rhizobacteria, the Pf.SS101-induced resistance response to Pst is dependent on salicylic acid signaling and not on jasmonic acid and ethylene signaling. Genome-wide transcriptomic and untargeted metabolomic analyses showed that in roots and leaves of Arabidopsis plants treated with Pf.SS101, approximately 1,910 genes and 50 metabolites were differentially regulated relative to untreated plants. Integration of both sets of “omics” data pointed to a prominent role of camalexin and glucosinolates in the Pf.SS101-induced resistance response. Subsequent bioassays with seven Arabidopsis mutants (myb51, cyp79B2cyp79B3, cyp81F2, pen2, cyp71A12, cyp71A13, and myb28myb29) disrupted in the biosynthesis pathways for these plant secondary metabolites showed that camalexin and glucosinolates are indeed required for the induction of Pst resistance by Pf.SS101. Also for the insect S. exigua, the indolic glucosinolates appeared to play a role in the Pf.SS101-induced resistance response. This study provides, to our knowledge for the first time, insight into the substantial biochemical and temporal transcriptional changes in Arabidopsis associated with the salicylic acid-dependent resistance response induced by specific rhizobacteria. PMID:23073694

  16. Phenazine-1-Carboxylic Acid Production by Pseudomonas fluorescens LBUM636 Alters Phytophthora infestans Growth and Late Blight Development.

    Science.gov (United States)

    Morrison, Christopher K; Arseneault, Tanya; Novinscak, Amy; Filion, Martin

    2017-03-01

    Phytophthora infestans causes late blight of potato, one of the most devastating diseases affecting potato production. Alternative approaches for controlling late blight are being increasingly sought due to increasing environmental concerns over the use of chemical pesticides and the increasing resistance of P. infestans to fungicides. Our research group has isolated a new strain of Pseudomonas fluorescens (LBUM636) of biocontrol interest producing the antibiotic phenazine-1-carboxylic acid (PCA). Wild-type LBUM636 was shown to significantly inhibit the growth of Phytophthora infestans in in vitro confrontational assays whereas its isogenic mutant (phzC-; not producing PCA) only slightly altered the pathogen's growth. Wild-type LBUM636 but not the phzC- mutant also completely repressed disease symptom development on tubers. A pot experiment revealed that wild-type LBUM636 can significantly reduce P. infestans populations in the rhizosphere and in the roots of potato plants, as well as reduce in planta disease symptoms due to PCA production. The expression of eight common plant defense-related genes (ChtA, PR-1b, PR-2, PR-5, LOX, PIN2, PAL-2, and ERF3) was quantified in tubers, roots, and leaves by reverse-transcription quantitative polymerase chain reaction and revealed that the biocontrol observed was not associated with the induction of a plant defense response by LBUM636. Instead, a direct interaction between P. infestans and LBUM636 is required and PCA production appears to be a key factor for LBUM636's biocontrol ability.

  17. A novel psychrophilic lipase from Pseudomonas fluorescens with unique property in chiral resolution and biodiesel production via transesterification.

    Science.gov (United States)

    Luo, Yu; Zheng, Yitao; Jiang, Zhengbing; Ma, Yushu; Wei, Dongzhi

    2006-11-01

    A lipase-producing bacterium strain B68 screened from soil samples of China was identified as Pseudomonas fluorescens. With GenomeWalker, the open reading frame of lipase gene lipB68, encoding 476 amino acids, was cloned and expressed in Escherichia coli BL21 (DE3). By affinity chromatography, the recombinant LipB68 protein was purified to the purity of 95%. As a member of lipase subfamily I.3, LipB68 has a unique optimum temperature of 20 degrees C, which was the lowest in this subfamily. In chiral resolution, LipB68 effectively catalyzed the transesterification of both alpha-phenylethanol and alpha-phenylpropanol at 20 degrees C, achieving E values greater than 100 and 60 after 120 h, respectively. Among all the known catalysts in biodiesel production, LipB68 produced biodiesel with a yield of 92% after 12 h, at the lowest temperature of 20 degrees C, and is the first one of the I.3 lipase subfamily reported to be capable of catalyzing the transesterification reaction of biodiesel production. Since lipase-mediated biodiesel production is normally carried out at 35-50 degrees C, the availability of a highly active lipase with a low optimal temperature can provide substantial savings in energy consumption. Thus, this novel psychrophilic lipase (LipB68) may represent a highly competitive energy-saving biocatalyst.

  18. A novel psychrophilic lipase from Pseudomonas fluorescens with unique property in chiral resolution and biodiesel production via transesterification

    Energy Technology Data Exchange (ETDEWEB)

    Luo Yu; Zheng Yitao; Jiang Zhengbing; Ma Yushu; Wei Dongzhi [East China Univ. of Science and Tech., Shanghai (China). State Key Lab. of Bioreactor Engineering

    2006-11-15

    A lipase-producing bacterium strain B68 screened from soil samples of China was identified as Pseudomonas fluorescens. With GenomeWalker, the open reading frame of lipase gene lipB68, encoding 476 amino acids, was cloned and expressed in Escherichia coli BL21 (DE3). By affinity chromatography, the recombinant LipB68 protein was purified to the purity of 95%. As a member of lipase subfamily I.3, LipB68 has a unique optimum temperature of 20 C, which was the lowest in this subfamily. In chiral resolution, LipB68 effectively catalyzed the transesterification of both a-phenylethanol and a-phenylpropanol at 20 C, achieving E values greater than 100 and 60 after 120 h, respectively. Among all the known catalysts in biodiesel production, LipB68 produced biodiesel with a yield of 92% after 12 h, at the lowest temperature of 20 C, and is the first one of the I.3 lipase subfamily reported to be capable of catalyzing the transesterification reaction of biodiesel production. Since lipasemediated biodiesel production is normally carried out at 35-50 C, the availability of a highly active lipase with a low optimal temperature can provide substantial savings in energy consumption. Thus, this novel psychrophilic lipase (LipB68) may represent a highly competitive energy-saving biocatalyst. (orig.)

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

    African Journals Online (AJOL)

    PRECIOUS

    2009-12-15

    Dec 15, 2009 ... A protease producing bacteria was isolated from meat waste contaminated soil and identified as. Pseudomonas ... Key words: Alkaline protease, casein agar, meat waste contaminated soil, Pseudomonas fluorescens. INTRODUCTION ... advent of new frontiers in biotechnology, the spectrum of protease ...

  20. A ptsP deficiency in PGPR Pseudomonas fluorescens SF39a affects bacteriocin production and bacterial fitness in the wheat rhizosphere.

    Science.gov (United States)

    Godino, Agustina; Príncipe, Analía; Fischer, Sonia

    2016-04-01

    Pseudomonas fluorescens SF39a is a plant-growth-promoting bacterium isolated from wheat rhizosphere. In this report, we demonstrate that this native strain secretes bacteriocins that inhibit growth of phytopathogenic strains of the genera Pseudomonas and Xanthomonas. An S-type pyocin gene was detected in the genome of strain SF39a and named pys. A non-polar pys::Km mutant was constructed. The bacteriocin production was impaired in this mutant. To identify genes involved in bacteriocin regulation, random transposon mutagenesis was carried out. A miniTn5Km1 mutant, called P. fluorescens SF39a-451, showed strongly reduced bacteriocin production. This phenotype was caused by inactivation of the ptsP gene which encodes a phosphoenolpyruvate phosphotransferase (EI(Ntr)) of the nitrogen-related phosphotransferase system (PTS(Ntr)). In addition, this mutant showed a decrease in biofilm formation and protease production, and an increase in surface motility and pyoverdine production compared with the wild-type strain. Moreover, we investigated the ability of strain SF39a-451 to colonize the wheat rhizosphere under greenhouse conditions. Interestingly, the mutant was less competitive than the wild-type strain in the rhizosphere. To our knowledge, this study provides the first evidence of both the relevance of the ptsP gene in bacteriocin production and functional characterization of a pyocin S in P. fluorescens. Copyright © 2015 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.

  1. Draft genome sequences of seven 4-Formylaminooxyvinylglycine producers belonging to the Pseudomonas fluorescens species complex

    Science.gov (United States)

    Vinylglycines are non-proteinogenic amino acids that inhibit amino acid metabolism and ethylene production. In this report, we describe the draft genome sequences of seven isolates of Pseudomonas that produce 4-formylaminooxyvinylglycine, a compound known to inhibit the germination of grasses and t...

  2. 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

    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...

  3. Biocidal Activity of Formaldehyde and Nonformaldehyde Biocides toward Mycobacterium immunogenum and Pseudomonas fluorescens in Pure and Mixed Suspensions in Synthetic Metalworking Fluid and Saline

    Science.gov (United States)

    Selvaraju, Suresh B.; Khan, Izhar U. H.; Yadav, Jagjit S.

    2005-01-01

    The microbicidal activity of four different biocides was studied in synthetic metalworking fluid (MWF) against Mycobacterium immunogenum, a suspected causative agent for hypersensitivity pneumonitis, and Pseudomonas fluorescens, a representative for the predominant gram-negative bacterial contaminants of MWF. The results indicated that M. immunogenum is more resistant than P. fluorescens to the tested formaldehyde-releasing biocides (Grotan and Bioban), isothiazolone (Kathon), and phenolic biocide (Preventol). Kathon was effective against mycobacteria at lower concentrations than the other three test biocides in MWF. In general, there was a marked increase in biocidal resistance of both the test organisms when present in MWF matrix compared to saline. Increased resistance of the two test organisms to biocides was observed when they were in a mixed suspension (1:1 ratio). The results indicate the protective effect of the MWF matrix against the action of commonly used biocides on the MWF-colonizing microbial species of occupational health significance, including mycobacteria. PMID:15640232

  4. Survival of Escherichia coli o157:h7 co-cultured with different levels of pseudomonas fluorescens and lactobacillus plantarum on fresh beef

    Science.gov (United States)

    Tshabalala, P. A.; de Kock, H. L.; Buys, E. M.

    2012-01-01

    The purpose of this study was to investigate the effect of different levels of Pseudomonas fluorescens (102 and 106 log10 cfu/ml) and Lactobacillus plantarum (102 and 104 log10 cfu/ml) on the growth of Escherichia coli O157:H7 on beef loins. Beef loins inoculated with E. coli O157:H7 and P. fluorescens were aerobically stored for 7 days at 4 ºC, while those inoculated with E. coli O157:H7 and L. plantarum were vacuum packaged and stored for 8 weeks at 4 ºC. Aerobic Plate Counts (APC), E. coli O157:H7 and either P. fluorescens or L. plantarum counts were determined at different storage intervals. For the aerobically packaged beef loins, E. coli O157:H7 was detected throughout the 7 day storage period regardless of the P. fluorescens level in the inoculum. For the vacuum packaged beef loins, similar inoculum levels of E. coli O157:H7 and L. plantarum allowed E. coli O157:H7 to survive until week 5 of storage, while a higher inoculum level of L. plantarum inhibited E. coli O157:H7 from week 3. Once fresh beef has been contaminated with E. coli O157:H7, the level of P. fluorescens in the background flora does not inhibit its survival and growth. However, under vacuum storage, the application of L. plantarum as a biopreservative inhibits the survival of E. coli O157:H7 on beef. The higher the level of L. plantarum in the system, the earlier the onset of the inhibition. Farmers and abattoirs have to strengthen preventive strategies to eliminate contamination of beef carcasses with E. coli O157:H7. PMID:24031970

  5. Toluene promotes lid 2 interfacial activation of cold active solvent tolerant lipase from Pseudomonas fluorescens strain AMS8.

    Science.gov (United States)

    Yaacob, Norhayati; Mohamad Ali, Mohd Shukuri; Salleh, Abu Bakar; Rahman, Raja Noor Zaliha Raja Abdul; Leow, Adam Thean Chor

    2016-07-01

    The utilization of cold active lipases in organic solvents proves an excellent approach for chiral synthesis and modification of fats and oil due to the inherent flexibility of lipases under low water conditions. In order to verify whether this lipase can function as a valuable synthetic catalyst, the mechanism concerning activation of the lid and interacting solvent residues in the presence of organic solvent must be well understood. A new alkaline cold-adapted lipase, AMS8, from Pseudomonas fluorescens was studied for its structural adaptation and flexibility prior to its exposure to non-polar, polar aprotic and protic solvents. Solvents such as ethanol, toluene, DMSO and 2-propanol showed to have good interactions with active sites. Asparagine (Asn) and tyrosine (Tyr) were key residues attracted to solvents because they could form hydrogen bonds. Unlike in other solvents, Phe-18, Tyr-236 and Tyr-318 were predicted to have aromatic-aromatic side-chain interactions with toluene. Non-polar solvent also was found to possess highest energy binding compared to polar solvents. Due to this circumstance, the interaction of toluene and AMS8 lipase was primarily based on hydrophobicity and molecular recognition. The molecular dynamic simulation showed that lid 2 (residues 148-167) was very flexible in toluene and Ca(2+). As a result, lid 2 moves away from the catalytic areas, leaving an opening for better substrate accessibility which promotes protein activation. Only a single lid (lid 2) showed the movement following interactions with toluene, although AMS8 lipase displayed double lids. The secondary conformation of AMS8 lipase that was affected by toluene observed a reduction of helical strands and increased coil structure. Overall, this work shows that cold active lipase, AMS8 exhibits distinguish interfacial activation and stability in the presence of polar and non-polar solvents. Copyright © 2016. Published by Elsevier Inc.

  6. Pseudomonas fluorescens F113 Mutant with Enhanced Competitive Colonization Ability and Improved Biocontrol Activity against Fungal Root Pathogens ▿

    Science.gov (United States)

    Barahona, Emma; Navazo, Ana; Martínez-Granero, Francisco; Zea-Bonilla, Teresa; Pérez-Jiménez, Rosa María; Martín, Marta; Rivilla, Rafael

    2011-01-01

    Motility is one of the most important traits for efficient rhizosphere colonization by Pseudomonas fluorescens F113rif (F113). In this bacterium, motility is a polygenic trait that is repressed by at least three independent pathways, including the Gac posttranscriptional system, the Wsp chemotaxis-like pathway, and the SadB pathway. Here we show that the kinB gene, which encodes a signal transduction protein that together with AlgB has been implicated in alginate production, participates in swimming motility repression through the Gac pathway, acting downstream of the GacAS two-component system. Gac mutants are impaired in secondary metabolite production and are unsuitable as biocontrol agents. However, the kinB mutant and a triple mutant affected in kinB, sadB, and wspR (KSW) possess a wild-type phenotype for secondary metabolism. The KSW strain is hypermotile and more competitive for rhizosphere colonization than the wild-type strain. We have compared the biocontrol activity of KSW with those of the wild-type strain and a phenotypic variant (F113v35 [V35]) which is hypermotile and hypercompetitive but is affected in secondary metabolism since it harbors a gacS mutation. Biocontrol experiments in the Fusarium oxysporum f. sp. radicis-lycopersici/Lycopersicum esculentum (tomato) and Phytophthora cactorum/Fragaria vesca (strawberry) pathosystems have shown that the three strains possess biocontrol activity. Biocontrol activity was consistently lower for V35, indicating that the production of secondary metabolites was the most important trait for biocontrol. Strain KSW showed improved biocontrol compared with the wild-type strain, indicating that an increase in competitive colonization ability resulted in improved biocontrol and that the rational design of biocontrol agents by mutation is feasible. PMID:21685161

  7. Pseudomonas fluorescens F113 mutant with enhanced competitive colonization ability and improved biocontrol activity against fungal root pathogens.

    Science.gov (United States)

    Barahona, Emma; Navazo, Ana; Martínez-Granero, Francisco; Zea-Bonilla, Teresa; Pérez-Jiménez, Rosa María; Martín, Marta; Rivilla, Rafael

    2011-08-01

    Motility is one of the most important traits for efficient rhizosphere colonization by Pseudomonas fluorescens F113rif (F113). In this bacterium, motility is a polygenic trait that is repressed by at least three independent pathways, including the Gac posttranscriptional system, the Wsp chemotaxis-like pathway, and the SadB pathway. Here we show that the kinB gene, which encodes a signal transduction protein that together with AlgB has been implicated in alginate production, participates in swimming motility repression through the Gac pathway, acting downstream of the GacAS two-component system. Gac mutants are impaired in secondary metabolite production and are unsuitable as biocontrol agents. However, the kinB mutant and a triple mutant affected in kinB, sadB, and wspR (KSW) possess a wild-type phenotype for secondary metabolism. The KSW strain is hypermotile and more competitive for rhizosphere colonization than the wild-type strain. We have compared the biocontrol activity of KSW with those of the wild-type strain and a phenotypic variant (F113v35 [V35]) which is hypermotile and hypercompetitive but is affected in secondary metabolism since it harbors a gacS mutation. Biocontrol experiments in the Fusarium oxysporum f. sp. radicis-lycopersici/Lycopersicum esculentum (tomato) and Phytophthora cactorum/Fragaria vesca (strawberry) pathosystems have shown that the three strains possess biocontrol activity. Biocontrol activity was consistently lower for V35, indicating that the production of secondary metabolites was the most important trait for biocontrol. Strain KSW showed improved biocontrol compared with the wild-type strain, indicating that an increase in competitive colonization ability resulted in improved biocontrol and that the rational design of biocontrol agents by mutation is feasible.

  8. 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.

  9. Xylanase B and an arabinofuranosidase from Pseudomonas fluorescens subsp. cellulosa contain identical cellulose-binding domains and are encoded by adjacent genes.

    OpenAIRE

    Kellett, L E; Poole, D M; Ferreira, L M; Durrant, A J; Hazlewood, G P; Gilbert, H J

    1990-01-01

    The complete nucleotide sequence of the Pseudomonas fluorescens subsp. cellulosa xynB gene, encoding an endo-beta-1,4-xylanase (xylanase B; XYLB) has been determined. The structural gene consists of an open reading frame (ORF) of 1775 bp coding for a protein of Mr 61,000. A second ORF (xynC) of 1712 bp, which starts 148 bp downstream of xynB, encodes a protein, designated xylanase C (XYLC), of Mr 59,000. XYLB hydrolyses oat spelt xylan to xylobiose and xylose, whereas XYLC releases only arabi...

  10. Characterization of an Endoglucanase from Pseudomonas fluorescens subsp. cellulosa Produced in Escherichia coli and Regulation of the Expression of Its Cloned Gene.

    OpenAIRE

    Lejeune, André; Courtois, Stéphane; Colson, Charles

    1988-01-01

    Several enzymatic properties of an endoglucanase produced in Escherichia coli by a gene from Pseudomonas fluorescens subsp. cellulosa were investigated. Gel filtration revealed a single peak of M(r) 36,000 with endoglucanase activity. The pH optimum of the enzyme was 7.0. Carboxymethyl cellulose and barley beta-glucan (mixed beta-1,3 and 1,4 linkages) were good substrates, but not laminarin (beta-1,3 linkages), amylose, filter paper, microcrystalline cellulose (Avicel), or cellotriose. The mo...

  11. 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.

  12. Quantitative and qualitative variability of the caseinolytic potential of different strains of Pseudomonas fluorescens: implications for the stability of casein micelles of UHT milks during their storage.

    Science.gov (United States)

    Baglinière, François; Tanguy, Gaëlle; Jardin, Julien; Matéos, Aurélie; Briard, Valérie; Rousseau, Florence; Robert, Benoît; Beaucher, Eric; Humbert, Gérard; Dary, Annie; Gaillard, Jean Luc; Amiel, Caroline; Gaucheron, Frédéric

    2012-12-15

    Pseudomonas fluorescens grows at low temperature and produces thermo-resistant protease(s) that can destabilize UHT (Ultra High Temperature) milk during its storage. The consequences of contamination of microfiltered milk with 9 strains of P. fluorescens on the stability of the corresponding UHT milk during storage had been investigated in this study. The strains were classified in two groups according to their ability to destabilize UHT milk. For the group of highly destabilizing strains, sedimentations of UHT milks, low values to phosphate test and the presence of aggregates were observed. Zeta potential and hydration of casein micelles decreased, whereas non casein nitrogen (NCN) and non protein nitrogen (NPN) contents increased. The analyses of NCN fraction by liquid chromatography coupled to mass spectrometry indicated that the different casein molecules were hydrolyzed in a similar way for the destabilizing strains suggesting that the same enzyme was implicated. For the group of slightly or not destabilizing strains no visual and biochemical alteration were found. This study showed that destabilization of UHT milk by P. fluorescens was highly variable and strain-dependent. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. Colonization process of olive tissues by Verticillium dahliae and its in planta interaction with the biocontrol root endophyte Pseudomonas fluorescens PICF7

    Science.gov (United States)

    Prieto, Pilar; Navarro‐Raya, Carmen; Valverde‐Corredor, Antonio; Amyotte, Stefan G.; Dobinson, Katherine F.; Mercado‐Blanco, Jesús

    2009-01-01

    Summary The colonization process of Olea europaea by the defoliating pathotype of Verticillium dahliae, and the in planta interaction with the endophytic, biocontrol strain Pseudomonas fluorescens PICF7 were determined. Differential fluorescent protein tagging was used for the simultaneous visualization of P. fluorescens PICF7 and V. dahliae in olive tissues. Olive plants were bacterized with PICF7 and then transferred to V. dahliae‐infested soil. Monitoring olive colonization events by V. dahliae and its interaction with PICF7 was conducted using a non‐gnotobiotic system, confocal laser scanner microscopy and tissue vibratoming sections. A yellow fluorescently tagged V. dahliae derivative (VDAT‐36I) was obtained by Agrobacterium tumefaciens‐mediated transformation. Isolate VDAT‐36I quickly colonized olive root surface, successfully invaded root cortex and vascular tissues via macro‐ and micro‐breakages, and progressed to the aerial parts of the plant through xylem vessel cells. Strain PICF7 used root hairs as preferred penetration site, and once established on/in root tissues, hindered pathogen colonization. For the first time using this approach, the entire colonization process of a woody plant by V. dahliae is reported. Early and localized root surface and root endophytic colonization by P. fluorescens PICF7 is needed to impair full progress of verticillium wilt epidemics in olive. PMID:21255281

  14. 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.

  15. 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.

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

    Science.gov (United States)

    2012-02-01

    contribute to the degradation of those coatings. Specifically, we characterized how medium conditions and substrate composition contribute to growth of...interfaces (Brown et al. 2010; Branda et al. 2005). Pseudomonad bacteria have been shown to be common constituents of aviation fuel microbiota in...These results demonstrate that medium composition impacts both the biofilm and cell populations, but not in the same way. In addition, carbon is

  17. 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.

  18. Factors screening to statistical experimental design of racemic atenolol kinetic resolution via transesterification reaction in organic solvent using free Pseudomonas fluorescens lipase.

    Science.gov (United States)

    Agustian, Joni; Kamaruddin, Azlina Harun; Aboul-Enein, Hassan Y

    2017-07-01

    As the (R)-enantiomer of racemic atenolol has no β-blocking activity and no lack of side effects, switching from the racemate to the (S)-atenolol is more favorable. Transesterification of racemic atenolol using free enzymes investigated as a resource to resolve the racemate via this method is limited. Screenings of enzyme, medium, and acetyl donor were conducted first to give Pseudomonas fluorescens lipase, tetrahydrofuran, and vinyl acetate. A statistical design of the experiment was then developed using Central Composite Design on some operational factors, which resulted in the conversions of 11.70-61.91% and substrate enantiomeric excess (ee) of 7.31-100%. The quadratic models are acceptable with R 2 of 95.13% (conversion) and 89.63% (ee). The predicted values match the observed values reasonably well. Temperature, agitation speed, and substrate molar ratio factor have low effects on conversion and ee, but enzyme loading affects the responses highly. The interaction of temperature-agitation speed and temperature-substrate molar ratio show significant effects on conversion, while temperature-agitation speed, temperature-substrate molar ratio, and agitation speed-substrate molar ratio affect ee highly. Optimum conditions for the use of Pseudomonas fluorescens lipase, tetrahydrofuran, and vinyl acetate were found at 45°C, 175 rpm, 2000 U, and 1:3.6 substrate molar ratio. © 2017 Wiley Periodicals, Inc.

  19. Multistep introduction of bacteria to natural substrates at different initial inoculation doses

    NARCIS (Netherlands)

    Kupriyanov, A.A.; Semenov, A.M.; Kunenkova, N.N.; Bruggen, van A.H.C.

    2009-01-01

    The population dynamics of the saprotrophic Pseudomonas fluorescens 32 gfp bacteria and two conventionally pathogenic enterobacteria (Escherichia coli 0157:H7 and Salmonella enterica var. Typhimurium) were investigated in their inoculation at different doses into cattle excreta and their subsequent

  20. Unraveling root developmental programs initiated by beneficial Pseudomonas spp. bacteria

    NARCIS (Netherlands)

    Zamioudis, C.; Mastranesti, P.; Dhonukshe, P.; Blilou, I.; Pieterse, C.M.J.

    2013-01-01

    Plant roots are colonized by an immense number of microbes, referred to as the root microbiome. Selected strains of beneficial soil-borne bacteria can protect against abiotic stress and prime the plant immune system against a broad range of pathogens. Pseudomonas spp. rhizobacteria represent one of

  1. Unraveling Root Developmental Programs Initiated by Beneficial Pseudomonas spp. Bacteria

    NARCIS (Netherlands)

    Zamioudis, C.; Mastranesti, P.; Dhonukshe, P.; Blilou, I.; Pieterse, C.M.J.

    2013-01-01

    Plant roots are colonized by an immense number of microbes, referred to as the root microbiome. Selected strains of beneficial soil-borne bacteria can protect against abiotic stress and prime the plant immune system against a broad range of pathogens. Pseudomonas spp. rhizobacteria represent one of

  2. In vitro inoculation of Aeromonas hydrophila and Pseudomonas fluorescens in cryopreserved silver barb (Barbodes gonionotus) milt: Effect on fertilization capacity and transmission potential to embryos.

    Science.gov (United States)

    Boonthai, Traimat; Khaopong, Weerasith; Sangsong, Jumlong; Vuthiphandchai, Verapong; Nimrat, Subuntith

    2017-11-16

    Purposive use of cryopreserved sperm contaminated with pathogenic agents has increased the risk of spreading of fish diseases. The objective of this study was to investigate the effects of A. hydrophila subsp. hydrophila and P. fluorescens inoculations into cryostored milt on fertilization capacity and transmission potential to embryos of silver barb (Barbodes gonionotus) with or without 0.25% penicillin-streptomycin (PS) administration. The experiment comprised six treatments: addition of milt into T1) extender only, T2) extender with 0.25% PS, T3) extender with A. hydrophila subsp. hydrophila (BG19), T4) extender with A. hydrophila subsp. hydrophila (BG19) and 0.25% PS, T5) extender with P. fluorescens (BG20) and T6) extender with P. fluorescens (BG20) and 0.25% PS. Milt were loaded into 0.25-mL straws and cryostored in the controlled-rate programmable freezer. After a cryostorage for 28 d, post-thawed sperm were evaluated for the fertilization capacity and risk of pathogen transmission to embryos. Inoculation of A. hydrophila subsp. hydrophila and P. fluorescens into extended milt (T3 and T5) caused a reduction (P fertilization capacity of cryopreserved sperm. Cryopreserved sperm inoculated with the two pathogenic bacteria and 0.25% PS (T4 and T6) did not fertilize the eggs. The two pathogenic bacteria could be transmitted into embryos after artificial insemination of eggs with bacterial-inoculated cryopreserved sperm, suggesting that the risks of disease transmission via cryopreserved fish sperm would exist. This is the first study reporting pathogenic bacterial transmission on in vitro fish embryos through artificial insemination of cryopreserved sperm. Copyright © 2017. Published by Elsevier Inc.

  3. 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

  4. Removal of aldrin, dieldrin, heptachlor, and heptachlor epoxide using activated carbon and/or Pseudomonas fluorescens free cell cultures.

    Science.gov (United States)

    Bandala, Erick R; Andres-Octaviano, Juan; Pastrana, Paulino; Torres, Luis G

    2006-01-01

    Degradation of aldrin (1,2,3,4,10,10-Hexachloro-1,4,4a,5,8,8a-hexahydro-1,4:5-8-dimethanonaphthalene), heptachlor (1H-1,4,5,6,7,8,8-heptachloro-3a,4,7,7a-tetrahydro-4,7-methano indene), dieldrin (1aalpha,2beta,2aalpha,3beta,6beta,6aalpha,7beta,7aalpha)-3,4,5,6,9,9-Hexachloro-1a,2,2a,3,6,6a,7,7a-octahydro-2,7:3,6-d-methanonaphtha[2,3-b]oxirene, and heptachlor epoxide (1aalpha, 1bbeta,2alpha,5alpha,5alphabeta,6beta,6aalpha-2,3,4,5,6,7,7-Heptachloro-1a,1b,5,5a,6,6a-hexahydro-2,5-methano-2H-inden[1,2-b]-oxirene) was tested using free cultures of Pseudomonas fluorescens under controlled conditions. Pesticide concentrations were monitored by gas chromatography during 120 h. Percentages of degradation and biodegradation rates (BDR) were calculated. Data showed a trend suggesting a relation between chemical structure and degradability. Degradation kinetics for each pesticide tested showed that the highest degradation rates were found in the first 24 h. Kinetics data were adjusted to an empirical equation in order to predict their behavior, and the correlation coefficients obtained were satisfactory. Gas chromatography/mass spectrometry (GC/MS) analysis of the final extracts allowed the identification of chlordene and monodechlorodieldrin, which have been reported as final metabolite produced in the biodegradation of this kind of compounds. Regarding adsorption of pesticides on activated vegetal carbon, we concluded that removal efficiencies between 95.45 and 97.18% can be reached, depending on the pesticide and the carbon dose applied. The values for K from the Freundlich equation were quite similar for the four pesticides (between 1.0001 and 1.04), whereas the n values were quite different for each pesticide in the following order of affinity: dieldrin > aldrin > heptachlor epoxide > heptachlor. Equilibrium times, very important for scaling up the process, were between 43 min and 1 h, for the heptachlor epoxide and the heptachlor, respectively.

  5. Genome Analysis of Pseudomonas fluorescens PCL1751: A Rhizobacterium that Controls Root Diseases and Alleviates Salt Stress for Its Plant Host.

    Directory of Open Access Journals (Sweden)

    Shu-Ting Cho

    Full Text Available Pseudomonas fluorescens PCL1751 is a rod-shaped Gram-negative bacterium isolated from the rhizosphere of a greenhouse-grown tomato plant in Uzbekistan. It controls several plant root diseases caused by Fusarium fungi through the mechanism of competition for nutrients and niches (CNN. This mechanism does not rely on the production of antibiotics, so it avoids the concerns of resistance development and is environmentally safe. Additionally, this bacterium promotes plant growth by alleviating salt stress for its plant host. To investigate the genetic mechanisms that may explain these observations, we determined the complete genome sequence of this bacterium, examined its gene content, and performed comparative genomics analysis with other Pseudomonas strains. The genome of P. fluorescens PCL1751 consisted of one circular chromosome that is 6,143,950 base-pairs (bp in size; no plasmid was found. The annotation included 19 rRNA, 70 tRNA, and 5,534 protein-coding genes. The gene content analysis identified a large number of genes involved in chemotaxis and motility, colonization of the rhizosphere, siderophore biosynthesis, and osmoprotectant production. In contrast, the pathways involved in the biosynthesis of phytohormones or antibiotics were not found. Comparison with other Pseudomonas genomes revealed extensive variations in their genome size and gene content. The presence and absence of secretion system genes were highly variable. As expected, the synteny conservation among strains decreased as a function of phylogenetic divergence. The integration of prophages appeared to be an important driver for genome rearrangements. The whole-genome gene content analysis of this plant growth-promoting rhizobacterium (PGPR provided some genetic explanations to its phenotypic characteristics. The extensive and versatile substrate utilization pathways, together with the presence of many genes involved in competitive root colonization, provided further support

  6. Peudomonas fluorescens diversity and abundance in the rhizosphere

    Science.gov (United States)

    Amina, Melinai; Ahmed, Bensoltane; Khaladi, Mederbel

    2010-05-01

    It is now over 30 years since that a several plant associated strains of fluorescent Pseudomonas spp. are known to produce antimicrobial metabolites, playing a significant role in the biological control of a lot of plant diseases. For that, the interest in the use of these bacteria for biocontrol of plant pathogenic agents has increased. However, few comprehensive studies have described the abundance of this soil borne bacteria in the region of Mascara (Northern-Algerian West). In the connection of this problem, this work was done by monitoring the number of indigenous Pseudomonas fluorescens organisms in three stations characterizing different ecosystems, to document their abundance, diversity and investigate the relationship between P. fluorescens abundance and soil properties. Our quantitative plate counting results hence the conception of their ecology in the rhizosphere. Thus, quantitative results has confirmed that P. fluorescens are successful root colonizers with strong predominance and competed for many ecological niche, where their distribution were correlated significantly (P<0.05) with the majority of soil properties. Keywords: P. Fluorescens, Ecosystems, Abundance, Diversity, Correlated, Soil Properties.

  7. Kynureninase-Type enzymes of Penicillum roqueforti, Aspergillus niger, Rhizopus stolonifer, and Pseudomonas fluorescens: further evidence for distinct kynureninase and hydroxykynureninase activities.

    Science.gov (United States)

    Shetty, A S; Gaertner, F H

    1975-04-01

    The kynureninase-type enzymes of three fungi and one bacterium were isolated and examined kinetically for their ability to catalyze the hydrolysis of L-kynurenine and L-3-hydroxykynurenine. The phycomycete Rhizopus stolonifer was found to contain a single, constitutive enzyme with Km for L-3-hydroxykynurenine and L-kynurenine of 6.67 times 10-minus 6 and 2.5 times 10-minus 4 M, respectively. The ascomycetes Aspergillus niger and Penicillium roqueforti each contain an enzyme, induced by L-tryptophan, with similar Km for L-3-hydroxykynurenine and L-kynurenine ranging from 5.9 times 10-minus 5 to 14.3 times 10-minus 5 M, as well as a constitutive enzyme with Km for the two substrates of similar to 4 times 10-minus 6 M and 10-minus 4 M. The bacterium Pseudomonas fluorescens has a single, inducible enzyme with Km for L-3-hydroxykynurenine and L-kynurenine of 5 times 10-minus 4 and 7 times 10-minus 5 M. In addition, significant differences in maximal velocities (Vmax) were observed in two cases. The Vmax of the inducible activity from P. fluorescens was 4.5 times greater for L-kynurenine than L-3-hydroxykynurenine, whereas the Vmax of the constitutive activity from R. stolonifer was 2.5 times greater for L-3-hydroxykynurenine. It is concluded (i) that the constitutive activities are hydroxykynureninases involved in the biosynthesis of nicotinamide adenine dinucleotide from L-tryptophan, (ii) that the inducible activities are kynureninases involved in the catabolism of L-tryptophan to anthranilate, and (iii) that R. stolonifer and P. fluorescens, respectively, carry the most specific examples of each type of enzyme.

  8. 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.

  9. Xylanase B and an arabinofuranosidase from Pseudomonas fluorescens subsp. cellulosa contain identical cellulose-binding domains and are encoded by adjacent genes.

    Science.gov (United States)

    Kellett, L E; Poole, D M; Ferreira, L M; Durrant, A J; Hazlewood, G P; Gilbert, H J

    1990-12-01

    The complete nucleotide sequence of the Pseudomonas fluorescens subsp. cellulosa xynB gene, encoding an endo-beta-1,4-xylanase (xylanase B; XYLB) has been determined. The structural gene consists of an open reading frame (ORF) of 1775 bp coding for a protein of Mr 61,000. A second ORF (xynC) of 1712 bp, which starts 148 bp downstream of xynB, encodes a protein, designated xylanase C (XYLC), of Mr 59,000. XYLB hydrolyses oat spelt xylan to xylobiose and xylose, whereas XYLC releases only arabinose from the same substrate. Thus XYLB is a typical xylanase and XYLC is an arabinofuranosidase. Both enzymes bind to crystalline cellulose (Avicel), but not to xylan. The nucleotide sequences between residues 114 and 931 of xynB and xynC were identical, as were amino acid residues 39-311 of XYLB and XYLC. This conserved sequence is reiterated elsewhere in the P. fluorescens subsp. cellulosa genome. Truncated derivatives of XYLB and XYLC, in which the conserved sequence had been deleted, retained catalytic activity, but did not exhibit cellulose binding. A hybrid gene in which the 5' end of xynC, encoding residues 1-110 of XYLC, was fused to the Escherichia coli pho A' gene (encodes mature alkaline phosphatase) directed the synthesis of a fusion protein which exhibited alkaline phosphatase activity and bound to cellulose.

  10. Characterization of an Endoglucanase from Pseudomonas fluorescens subsp. cellulosa Produced in Escherichia coli and Regulation of the Expression of Its Cloned Gene.

    Science.gov (United States)

    Lejeune, A; Courtois, S; Colson, C

    1988-02-01

    Several enzymatic properties of an endoglucanase produced in Escherichia coli by a gene from Pseudomonas fluorescens subsp. cellulosa were investigated. Gel filtration revealed a single peak of M(r) 36,000 with endoglucanase activity. The pH optimum of the enzyme was 7.0. Carboxymethyl cellulose and barley beta-glucan (mixed beta-1,3 and 1,4 linkages) were good substrates, but not laminarin (beta-1,3 linkages), amylose, filter paper, microcrystalline cellulose (Avicel), or cellotriose. The mode of action was typical of an "endo"-acting enzyme. Taken together, these properties do not correspond to those of any of the endoglucanases described in P. fluorescens subsp. cellulosa. Consequently, the gene was designated egIX. The enzyme was sensitive to end-product inhibition by cellobiose but was only moderately inhibited by glucose. The enzyme was formed constitutively in E. coli throughout the growth phase. Urea had no effect on endoglucanase synthesis, but glucose acted as a catabolite repressor. The formation of the enzyme in E. coli was partially dependent on cyclic AMP.

  11. The N-terminal region of an endoglucanase from Pseudomonas fluorescens subspecies cellulosa constitutes a cellulose-binding domain that is distinct from the catalytic centre.

    Science.gov (United States)

    Gilbert, H J; Hall, J; Hazlewood, G P; Ferreira, L M

    1990-05-01

    The substrate specificity of an endoglucanase (EGB) from Pseudomonas fluorescens subspecies cellulosa was determined. The enzyme was most active against barley beta-glucan, but showed significant activity against amorphous and crystalline cellulose. EGB was purified to homogeneity by affinity chromatography with crystalline cellulose (Avicel). The Mr of the purified enzyme was 50,000, which is in good agreement with the size of EGB deduced from the nucleotide sequence of the celB gene, coding for EGB. The N-terminal region of the mature form of EGB showed strong homology to another endoglucanase and to a xylanase expressed by the same organism; homologous sequences included highly conserved serine-rich regions. Truncated forms of celB, in which the gene sequence encoding the conserved domain had been deleted, directed the synthesis of a functional endoglucanase that did not bind to crystalline cellulose. This indicates that the conserved region of endoglucanases and xylanases expressed by P. fluorescens subsp. cellulosa constitutes a cellulose-binding domain, which is distinct from the active centre. The possible role of this substrate-binding region is discussed.

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

    Science.gov (United States)

    Maldonado-González, M. Mercedes; Bakker, Peter A. H. M.; Prieto, Pilar; Mercado-Blanco, Jesús

    2015-01-01

    The effective management of Verticillium wilts (VW), 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 VW 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 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 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. A. thaliana is therefore a suitable alternative to olive bioassays to unravel biocontrol traits involved in biological control of V. dahliae by P. fluorescens PICF7. PMID:25904904

  13. 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.

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

    DEFF Research Database (Denmark)

    Johansen, Anders; Knudsen, Inge M.B.; Binnerup, Svend 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...... 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...... 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...

  15. [Structural and physiological diversity among cystlike resting cells of bacteria of the genus Pseudomonas].

    Science.gov (United States)

    Muliukin, A L; Suzina, N E; Duda, V I; El'-Registan, G I

    2008-01-01

    Cystlike resting cells (CRC) of non-spore-forming gram-negative bacteria of the genus Pseudomonas, P. aurantiaca and P. fluorescens, were obtained and characterized for the first time; their physiological and morphological diversity was demonstrated. The following properties were common for all the revealed types of CRC as dormant forms: (1) long-term (up to 6 months or longer) maintenance of viability in the absence of culture growth and cell respiration; (2) absence of an experimentally detectable level of metabolism; (3) higher resistance to damage and autolysis under the action of provoking factors than in metabolically active vegetative cells; and (4) specific features of ultrastructural organization absent in vegetative cells: thickened and lamellar envelopes, clumpy structure of the cytoplasm, and condensed DNA in nucleoid. The differences in various types of CRC concern the thickness and lamellar structure of cell envelopes, as well as the presence and thickness of the capsular layer. In particular, forms ultrastructurally similar to typical bacterial cysts were revealed in pseudomonad populations growing on soil agar. Physiological diversity was revealed in different levels of viability preservation and thermal resistance in various types of CRC and depended on the conditions of their formation. The optimal conditions and procedures for obtaining P. aurantiaca and P. fluorescens CRC that retain the ability to form colonies on standard nutrient media are as follows: (1) a twofold decrease of nitrogen content in the growth medium; (2) an increased level of anabiosis autoinducer (C12-AHB, 10(-4) M) in stationary cultures; (3) transfer of the cells from stationary cultures to a starvation medium with silica; (4) cultivation in soil extract; and (5) development of cultures on soil agar. The CRC from the cultures grown in soil extract or starvation medium with silica proved to be resistant to heat treatment (60 degrees C, 5 min). In the CRC formed in nitrogen

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

    African Journals Online (AJOL)

    A protease producing bacteria was isolated from meat waste contaminated soil and identified as Pseudomonas fluorescens. Optimization of the fermentation medium for maximum protease production was carried out. The culture conditions like inoculum concentration, incubation time, pH, temperature, carbon sources, ...

  17. Efficiency of the formulated plant-growth promoting Pseudomonas fluorescens MC46 inoculant on triclocarban treatment in soil and its effect on Vigna radiata growth and soil enzyme activities.

    Science.gov (United States)

    Sipahutar, Merry Krisdawati; Piapukiew, Jittra; Vangnai, Alisa S

    2018-02-15

    For bioaugmentation-based treatment of triclocarban (TCC), an emerging soil pollutant that is recalcitrant to biodegradation and phytotransformation, efficient TCC-degrading bacteria with an effective soil-delivering means are required. This work developed the formulated bacterial inoculant, and successfully demonstrated its TCC removal and detoxification performance in pot soil experiment with Vigna radiata plants. The soil bacterium Pseudomonas fluorescens MC46 was isolated as TCC-degrading, plant-growth promoting bacterium. The characterizations were conducted in vitro revealing that it could utilize TCC as a sole carbon source, and at a wide and higher concentration range from 1.6-31.6mgkg -1 than those previously reported, while the detoxification was assessed by cytogenotoxicity and phytotoxicity tests. The developed sawdust-based inoculant formula combined with molasses (5% w/w), and either PEG or CMC-starch blend (1% w/w) could maintain a 20-week shelf-life inoculant stability in terms of cell viability, and TCC-degrading activity. Bioaugmentation of the formulated inoculants into TCC-contaminated soil efficiently removed TCC up to 74-76% of the initial concentration, mitigated toxicity, restored plant growth and health, and enhanced soil enzyme activities. This work is the first to demonstrate potential application of the formulated plant-growth promoting bacterial inoculant for the treatment and detoxification of a persistent TCC contaminated in soil. Copyright © 2017 Elsevier B.V. All rights reserved.

  18. Effect of Pseudomonas Sp. Bacteria on Soil Chemical and Biological Properties, Yield and Its Components of Two Rice Cultivars

    Directory of Open Access Journals (Sweden)

    Z. AminDeldar

    2014-04-01

    Full Text Available To evaluate the effect of plant growth promoting bacteria (PGPB on soil physical and chemical indices, yield and its components of two rice cultivars, an experiment was arranged in Rice Researches Institute of Guilan Province during 2009. The experiment design consisted of four randomized complete blocks in a factorial arrangement having 16 treatments in every block. In this research, two factors were evaluated: first, two cultivars (Khazar and Hashemi and second, eight levels of seed inoculation with PGPB (P.fluorescens strain 168, P.fluorescens strain 93, P.fluorescens strain 177, P.fluorescens strain 136, P.fluorescens strain 103, P.fluorescens strain 169, P.fluorescens strain 4 and control (without inoculation. Investigated characteristics consisted of: soil minerals, no.microorganisms in soil, grain yield, number of seed per panicle, number of seed per plant, 1000-seed weight, biological and economic yield. In this experiment, effect of cultivar and bacteria were significant in the most of studied characteristics, but effect of cultivar*bacteria (except yield components wasn’t significant. The results of experiment showed that inoculation with bacterial strains had a stimulating effect on growth and development of rice cultivars. In this experiment, Khazar had positive effect on the most of studied characteristics in compared with Hashemi. Between bacterial different strains, seed inoculation with 168, 177 and 93 strains in compared with other strains increased evaluated characteristics significantly. Seed inoculation with PGPB improved studied characteristics and microorganisms in soil, too.

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

    Directory of Open Access Journals (Sweden)

    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.

  20. Proteolytic activity of protease produced by Pseudomonas fluorescens IB 2312 in skimmed milk subject to the process of high pressure homogenization

    Directory of Open Access Journals (Sweden)

    Claudia Regina Gonçalves Pinho

    2014-08-01

    Full Text Available The presence of thermoresistant proteases produced by psychrotrophic microorganisms have been identified as a limiting factor of the UHT milk shelflife, causing undesirable changes in milk products. High pressure homogenization (HPH processing is a non-thermal method of food preservation, able to promotes the microbiological safety and inactivation of some enzymes. Thus, this work assessed the proteolytic activity of protease produced by Pseudomonas fluorescens in skim milk subjected to high pressure homogenization process. The milk samples were added by the protease enzymatic extract (10% v/v and subjected to pressures up to 300 MPa. The assays showed that pressures on the order of 300 MPa caused a 72.5% reduction in proteolytic activity. Therefore, the process at high pressures resulted in significant inactivation of this thermoresistent enzyme, which possibly favors the shelf-life extension of the UHT milk and also limits the yield and quality loss of cheeses due to undesirable sensory changes in flavor and texture caused by this enzyme.

  1. 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.

  2. 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.

  3. 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.

  4. Diversity and functional analysis of LuxR-type transcriptional regulators of cyclic lipopeptide biosynthesis in Pseudomonas fluorescens

    NARCIS (Netherlands)

    Bruijn, de I.; Raaijmakers, J.M.

    2009-01-01

    Cyclic lipopeptides (CLPs) are produced by many Pseudomonas species and have several biological functions, including a role in surface motility, biofilm formation, virulence, and antimicrobial activity. This study focused on the diversity and role of LuxR-type transcriptional regulators in CLP

  5. Opening Study on the Development of a New Biosensor for Metal Toxicity Based on Pseudomonas fluorescens Pyoverdine

    Directory of Open Access Journals (Sweden)

    Alessandro Chiadò

    2013-12-01

    Full Text Available To date, different kinds of biosensing elements have been used effectively for environmental monitoring. Microbial cells seem to be well-suited for this task: they are cheap, adaptable to variable field conditions and give a measurable response to a broad number of chemicals. Among different pollutants, heavy metals are still a major problem for the environment. A reasonable starting point for the selection of a biorecognition element to develop a biosensor for metals could be that of a microorganism that exhibits good mechanisms to cope with metals. Pseudomonads are characterized by the secretion of siderophores (e.g., pyoverdine, low-molecular weight compounds that chelate Fe3+ during iron starvation. Pyoverdine is easily detected by colorimetric assay, and it is suitable for simple online measurements. In this work, in order to evaluate pyoverdine as a biorecognition element for metal detection, the influence of metal ions (Fe3+, Cu2+, Zn2+, but also of temperature, pH and nutrients, on microbial growth and pyoverdine regulation has been studied in P. fluorescens. Each of these variables has been shown to influence the synthesis of siderophore: for instance, the lower the temperature, the higher the production of pyoverdine. Moreover, the concentration of pyoverdine produced in the presence of metals has been compared with the maximum allowable concentrations indicated in international regulations (e.g., 98/83/EC, and a correlation that could be useful to build a colorimetric biosensor has been observed.

  6. Protozoan growth rates on secondary-metabolite-producing Pseudomonas spp. correlate with high-level protozoan taxonomy

    DEFF Research Database (Denmark)

    Pedersen, Annette L.; Winding, Anne; Altenburger, Andreas

    2011-01-01

    Different features can protect bacteria against protozoan grazing, for example large size, rapid movement, and production of secondary metabolites. Most papers dealing with these matters focus on bacteria. Here, we describe protozoan features that affect their ability to grow on secondary......-metabolite-producing bacteria, and examine whether different bacterial secondary metabolites affect protozoa similarly. We investigated the growth of nine different soil protozoa on six different Pseudomonas strains, including the four secondary-metabolite-producing Pseudomonas fluorescens DR54 and CHA0, Pseudomonas...... chlororaphis MA342 and Pseudomonas sp. DSS73, as well as the two nonproducers P. fluorescens DSM50090T and P. chlororaphis ATCC43928. Secondary metabolite producers affected protozoan growth differently. In particular, bacteria with extracellular secondary metabolites seemed more inhibiting than bacteria...

  7. Protozoan-induced regulation of cyclic lipopeptide biosynthesis Is an effective predation defense mechanism for Pseudomonas fluorescens

    NARCIS (Netherlands)

    Mazzola, M.; Bruijn, de I.; Cohen, M.F.; Raaijmakers, J.M.

    2009-01-01

    Environmental bacteria are exposed to a myriad of biotic interactions that influence their function and survival. The grazing activity of protozoan predators significantly impacts the dynamics, diversification, and evolution of bacterial communities in soil ecosystems. To evade protozoan predation,

  8. The oxyanion hole of Pseudomonas fluorescens mannitol 2-dehydrogenase: a novel structural motif for electrostatic stabilization in alcohol dehydrogenase active sites.

    Science.gov (United States)

    Klimacek, Mario; Nidetzky, Bernd

    2009-12-23

    The side chains of Asn191 and Asn300 constitute a characteristic structural motif of the active site of Pseudomonas fluorescens mannitol 2-dehydrogenase that lacks precedent in known alcohol dehydrogenases and resembles the canonical oxyanion binding pocket of serine proteases. We have used steady-state and transient kinetic studies of the effects of varied pH and deuterium isotopic substitutions in substrates and solvent on the enzymatic rates to delineate catalytic consequences resulting from individual and combined replacements of the two asparagine residues by alanine. The rate constants for the overall hydride transfer to and from C-2 of mannitol, which were estimated as approximately 5 x 102 s-1 and approximately 1.5 x 103 s-1 in the wild-type enzyme respectively, were selectively slowed, between 540- and 2700-fold, in single-site mannitol 2-dehydrogenase mutants. These effects were additive in the corresponding doubly mutated enzyme, suggesting independent functioning of the two asparagine residues in catalysis. Partial disruption of the oxyanion hole in single-site mutants caused an upshift, by >or=1.2 pH units, in the kinetic pK of the catalytic acid-base Lys295 in the enzyme-NAD+-mannitol complex. The oxyanion hole of mannitol 2-dehydrogenase is suggested to drive a precatalytic conformational equilibrium at the ternary complex level in which the reactive group of the substrate is 'activated' for chemical conversion through its precise alignment with the unprotonated side chain of Lys295 (mannitol oxidation) and C=O bond polarization by the carboxamide moieties of Asn191 and Asn300 (fructose reduction). In the subsequent hydride transfer step, the two asparagine residues provide approximately 40 kJ/mol of electrostatic stabilization.

  9. Improvement of a dry formulation of Pseudomonas fluorescens EPS62e for fire blight disease biocontrol by combination of culture osmoadaptation with a freeze-drying lyoprotectant.

    Science.gov (United States)

    Cabrefiga, J; Francés, J; Montesinos, E; Bonaterra, A

    2014-10-01

    To study the effect of lyoprotectants and osmoadaptation on viability of Pseudomonas fluorescens EPS62e during freeze-drying and storage and to evaluate the formulation in terms of efficacy in biocontrol and fitness on pear flowers. A wettable powder formulation of a biocontrol agent of fire blight was optimized by means of lyoprotectants and culture osmoadaptation. Freeze-drying was used to obtain dehydrated cells, and the best viability (70% of survival) was obtained using lactose as lyoprotectant. Survival during lyophilization was additionally improved using physiological adaptation of cells during cultivation under salt-amended medium (osmoadaptation). The procedure increased the survival of cells after freeze-drying attaining viability values close to a 100% in the lactose-formulated product (3 × 10(11) CFU g(-1) ), and through the storage period of 1 year at 4°C. The dry formulation showed also an improved biocontrol efficacy and survival of EPS62e on pear flowers under low relative humidity conditions. Cell viability after freeze-drying was improved using lactose as lyoprotectant combined with a procedure of osmoadaptation during cultivation. The powder-formulated product remained active for 12 months and retained biocontrol levels similar to that of fresh cells. The formulation showed an improved survival of EPS62e on flowers and an increase of the efficacy of biocontrol of fire blight at low relative humidity. The results have a potential value for commercial application in biocontrol agents not only of fire blight but also of other plant diseases. © 2014 The Society for Applied Microbiology.

  10. 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

    OpenAIRE

    Avendaño, Camila; Arbeláez, Germán; Rondón, Guillermo

    2010-01-01

    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 Tibaitatá, Mosquera (Cundinamarca), utilizando un diseño co...

  11. Nitrogen availability to Pseudomonas fluorescens DF57 is limited during decomposition of barley straw in bulk soil and in the barley rhizosphere.

    Science.gov (United States)

    Jensen, L E; Nybroe, O

    1999-10-01

    The availability of nitrogen to Pseudomonas fluorescens DF57 during straw degradation in bulk soil and in barley rhizosphere was studied by introducing a bioluminescent reporter strain (DF57-N3), responding to nitrogen limitation, to model systems of varying complexity. DF57-N3 was apparently not nitrogen limited in the natural and sterilized bulk soil used for these experiments. The soil was subsequently amended with barley straw, representing a plant residue with a high carbon-to-nitrogen ratio (between 60 and 100). In these systems the DF57-N3 population gradually developed a nitrogen limitation response during the first week of straw decomposition, but exclusively in the presence of the indigenous microbial population. This probably reflects the restricted ability of DF57 to degrade plant polymers by hydrolytic enzymes. The impact of the indigenous population on nitrogen availability to DF57-N3 was mimicked by the cellulolytic organism Trichoderma harzianum Rifai strain T3 when coinoculated with DF57-N3 in sterilized, straw-amended soil. Limitation occurred concomitantly with fungal cellulase production, pointing to the significance of hydrolytic activity for the mobilization of straw carbon sources, thereby increasing the nitrogen demand. Enhanced survival of DF57-N3 in natural soil after straw amendment further indicated that DF57 was cross-fed with carbon/energy sources. The natural barley rhizosphere was experienced by DF57-N3 as an environment with restricted nitrogen availability regardless of straw amendment. In the rhizosphere of plants grown in sterilized soil, nitrogen limitation was less severe, pointing to competition with indigenous microorganisms as an important determinant of the nitrogen status for DF57-N3 in this environment. Hence, these studies have demonstrated that nitrogen availability and gene expression in Pseudomonas is intimately linked to the structure and function of the microbial community. Further, it was demonstrated that the

  12. Isolation of Crude Oil from Polluted Waters Using Biosurfactants Pseudomonas Bacteria: Assessment of Bacteria Concentration Effects

    Directory of Open Access Journals (Sweden)

    A. Khalifeh

    2013-04-01

    Full Text Available Biological decomposition techniques and isolation of environmental pollutions using biosurfactants bacteria are effective methods of environmental protection. Surfactants are amphiphilic compounds that are produced by local microorganisms and are able to reduce the surface and the stresses between surfaces. As a result, they will increase solubility, biological activity, and environmental decomposition of organic compounds. This study analyzes the effects of biosurfactants on crude oil recovery and its isolation using pseudomonas sea bacteria species. Preparation of biosurfactants was done in glass flasks and laboratory conditions. Experiments were carried out to obtain the best concentration of biosurfactants for isolating oil from water and destroying oil-in-water or water-in-oil emulsions in two pH ranges and four saline solutions of different concentrations. The most effective results were gained when a concentration of 0.1% biosurfactants was applied.

  13. 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

  14. Annual changes in bioactive contents and production in field-grown blackberry after inoculation with Pseudomonas fluorescens.

    Science.gov (United States)

    Ramos-Solano, B; Garcia-Villaraco, A; Gutierrez-Mañero, F J; Lucas, J A; Bonilla, A; Garcia-Seco, D

    2014-01-01

    The aim of this study was two-fold: first, to characterize blackberry fruits from Rubus sp. var. Lochness along the year, and secondly, to evaluate the ability of a Pseudomonas strain (N21.4) to improve fruit yield and quality under field conditions in production greenhouses throughout the year. The strain was root or leaf inoculated to blackberry plants and fruits were harvested in each season. Nutritional parameters, antioxidant potential and bioactive contents were determined; total fruit yield was recorded. Blackberries grown under short day conditions (autumn and winter) showed significantly lower °Brix values than fruits grown under long day conditions. Interestingly, an increase in fruit °Brix, relevant for quality, was detected after bacterial challenge, together with significant and sustained increases in total phenolics and flavonoids. Improvements in inoculated fruits were more evident from October through early March, when environmental conditions are worse. In summary, N21.4 is an effective agent to increase fruit quality and production along the year in blackberry; this is an environmentally friendly approach to increase fruit quality. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

  15. Zinc Improves Biocontrol of Fusarium Crown and Root Rot of Tomato by Pseudomonas fluorescens and Represses the Production of Pathogen Metabolites Inhibitory to Bacterial Antibiotic Biosynthesis.

    Science.gov (United States)

    Duffy, B K; Défago, G

    1997-12-01

    ABSTRACT Crown and root rot of tomato caused by Fusarium oxysporum f. sp. radicis-lycopersici is an increasing problem in Europe, Israel, Japan, and North America. The biocontrol agent Pseudomonas fluorescens strain CHA0 provides only moderate control of this disease. A one-time amendment of zinc EDTA at 33 mug of Zn(2+)/ml to hydroponic nutrient solution in soilless rockwool culture did not reduce disease when used alone, but did reduce disease by 25% in the presence of CHA0. In in vitro studies with the pathogen, zinc at concentrations as low as 10 mug/ml abolished production of the phytotoxin fusaric acid, a Fusarium pathogenicity factor, and increased production of microconidia over 100-fold, but reduced total biomass. Copper EDTA at 33 mug of Cu(2+)/ml had a similar effect as zinc on the pathogen in vitro; it reduced disease when used alone, and increased the biocontrol activity of CHA0 in soilless culture. Ammonium-molybdate neither improved the biocontrol activity of CHA0 nor affected production of fusaric acid or microconidia. Strain CHA0 did not degrade fusaric acid. Fusaric acid at concentrations as low as 0.12 mug/ml repressed production by CHA0 of the antibiotic 2,4-diacetylphloroglucinol, a key factor in the biocontrol activity of this strain. Production of pyoluteorin by CHA0 was also reduced, but production of hydrogen cyanide and protease was not affected, suggesting that fusaric acid affects biosynthesis at a regulatory level downstream of gacA and apdA genes. Fusaric acid did not affect the recovery of preformed antibiotics nor did it affect bacterial growth even at concentrations as high as 200 mug/ml. When microbial meta-bolite production was measured in the rockwool bioassay, zinc amendments reduced fusaric acid production and enhanced 2,4-diacetylphloro-glucinol production. We suggest that zinc, which did not alleviate the repression of antibiotic biosynthesis by fusaric acid, improved biocontrol activity by reducing fusaric acid production by

  16. Efficacy of spray –Dried Pseudomonas fluorescens, strain CL145A (Zequanox®), for controlling Zebra Mussels (Dreissena polymorpha) within Lake Minnetonka, MN enclosures

    Science.gov (United States)

    Luoma, James A.; Severson, Todd J.

    2016-01-01

    The efficacy of whole water column and subsurface applications of the biopesticide Zequanox®, a commercially prepared spray-dried powder formulation of Pseudomonas fluorescens (strain CL145A), were evaluated for controlling zebra mussels (Dreissena polymorpha) within 27-m2 enclosures in Lake Minnetonka (Deephaven, Minnesota). Five treatments consisting of (1) two whole water column Zequanox applications, (2) two subsurface Zequanox applications, and (3) an untreated control were completed on each of three independent treatment days during September 2014. The two types of samplers used in the study were (1) type 1 samplers, which were custom built multi-plate samplers (wood, perforated aluminum, and tile substrates) that were placed into Robinson’s Bay in June of 2013 to allow for natural colonization by zebra mussels, and (2) type 2 samplers, which consisted of zebra mussels adhering to perforated aluminum trays that were placed into mesh containment bags. One day prior to treatment, three individual samplers of each type were distributed to test enclosures and exposed to a randomly assigned treatment. Sampling to determine the zebra mussel biomass adhering to type 1 samplers and the survival assessments for zebra mussels contained in type 2 samplers were completed ~40 days after exposure. The zebra mussel biomass adhering to type 1 samplers and the survival of zebra mussels contained in type 2 samplers were significantly less in groups treated with the highest Zequanox concentrations and in groups that received whole water column applications than comparable groups treated with lower Zequanox concentrations and subsurface applications. However, standardization of biomass and survival results to the amount of Zequanox applied showed that the lower concentrations and subsurface applications were more cost efficient, with respect to product used, at reducing zebra mussel biomass and for inducing zebra mussel mortality. Although the subsurface application methods

  17. 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

  18. Phenazine-producing fluorescent Pseudomonas spp.: Diversity and biogeography in central Washington state

    Science.gov (United States)

    Strains of the rhizosphere bacterium Pseudomonas fluorescens produce redox-active phenazine antibiotics that suppress a wide variety of soilborne plant pathogens. Our laboratory recently detected these bacteria a population levels up to 106 colony-forming units (cfu) per gram of root (fresh weight)...

  19. Interacción sinérgica entre hongos formadores de micorrizas arbusculares - Pseudomonas fluorescens y su relación en la nutrición vegetal de fósforo / Synergic interaction between arbuscular mycorrhizal fungi and Pseudomonas fluorescens and its effect on plant phosphorus uptake

    OpenAIRE

    Ordoñez Castañeda, Yuli Marcela

    2009-01-01

    Ciertos microorganismos (género Pseudomonas) y Hongos Formadores de Micorrizas Arbusculares (HFMA) tienen la habilidad de incrementar el fósforo disponible para la planta y mejorar la productividad de los cultivos agícolas. El uso coordinado de especies seleccionadas, tanto de bacterias como de HFMA, se convierte en una alternativa interesante, donde las bacterias liberan fosfatos y el micelio extraradical (MER) de este tipo de hongos puede movilizarlo hacia las raíces de la planta a la cual ...

  20. Bioactive potential of symbiotic bacteria and fungi from marine ...

    African Journals Online (AJOL)

    Marine sponges are rich in microbial biota. In this study, totally four sponges namely Callyspongia diffusa, Hyattella Cribriformis, Sigmadocia carnosa, Spongia officininalis Var ceylonensis were collected and their associated bacteria and fungi were isolated. Among the microbes isolated, Pseudomonas fluorescens and ...

  1. A novel defence pathway in Arabidopsis induced by biocontrol bacteria

    NARCIS (Netherlands)

    Pieterse, C.M.J.; Wees, A.C.M. van; Pelt, J.A. van; Loon, L.C. van

    1998-01-01

    Plants have the ability to acquire an enhanced level of resistance to pathogen attack after being exposed to specific biotic stimuli. In Arabidopsis thaliana, non-pathogenic, root-colonising Pseudomonas fluorescens bacteria with biological disease control activity trigger an induced systemic

  2. Effects of the tomato pathogen Fusarium oxysporum f. sp. radicis-lycopersici and of the biocontrol bacterium Pseudomonas fluorescens WCS365 on the composition of organic acids and sugars in tomato root exudate.

    Science.gov (United States)

    Kamilova, Faina; Kravchenko, Lev V; Shaposhnikov, Alexander I; Makarova, Nataliya; Lugtenberg, Ben

    2006-10-01

    The effects of the pathogenic fungus Fusarium oxysporum f. sp. radicis-lycopersici and of the bacterial biocontrol strain Pseudomonas fluorescens WCS365, and of both microbes, on the amounts and composition of root exudate components of tomato plants grown in a gnotobiotic stonewool substrate system were studied. Conditions were selected under which introduction of F. oxysporum f. sp. radicis-lycopersici caused severe foot and root rot, whereas inoculation of the seed with P. fluorescens WCS365 decreased the percentage of diseased plants from 96 to 7%. This is a much better disease control level than was observed in potting soil. Analysis of root exudate revealed that the presence of F. oxysporum f. sp. radicis-lycopersici did not alter the total amount of organic acids, but that the amount of citric acid decreased and that of succinic acid increased compared with the nontreated control. In contrast, in the presence of the P. fluorescens biocontrol strain WCS365, the total amount of organic acid increased, mainly due to a strong increase of the amount of citric acid, whereas the amount of succinic acid decreased dramatically. Under biocontrol conditions, when both microbes are present, the content of succinic acid decreased and the level of citric acid was similar to that in the nontreated control. The amount of sugar was approximately half that of the control sample when either one of the microbes was present alone or when both were present. Analysis of the interactions between the two microbes grown together in sterile tomato root exudate showed that WCS365 inhibited multiplication of F. oxysporum f. sp. radicis-lycopersici, whereas the fungus did not affect the number of CFU of the bacterium.

  3. Two strains of Pseudomonas fluorscens bacteria differentially affect survivorship of waxworm (Galleria mellonella) larvae exposed to an arthropod fungal pathogen, Beauveria bassiana

    Science.gov (United States)

    Two strains of Pseudomonas fluorescens were found contaminating a biopesticide used in a previous study against Varroa destructor infestations in honey bee hives. In the aforementioned study the biopesticide, a formulation of the arthropod pathogen Beauveria bassiana, failed to have any impact on t...

  4. 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.

    suppressive soil, P. fluorescens In5 is therefore a promising potential biocontrol agent with potent activity against plant pathogens. Studies to date have shown nunamycin and nunapeptin as key compounds underpinning this antimicrobial activity. A combination of molecular genetic strain manipulations and omic...

  5. 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.

  6. Carvacrol and 1,8-cineole alone or in combination at sublethal concentrations induce changes in the cell morphology and membrane permeability of Pseudomonas fluorescens in a vegetable-based broth.

    Science.gov (United States)

    de Sousa, Jossana Pereira; Torres, Rayanne de Araújo; de Azerêdo, Geíza Alves; Figueiredo, Regina Célia Bressan Queiroz; Vasconcelos, Margarida Angélica da Silva; de Souza, Evandro Leite

    2012-08-01

    This study aimed to investigate the effects of sublethal concentrations of carvacrol (CAR) and 1,8-cineole (CIN) alone and in combination on the morphology, cell viability and membrane permeability of Pseudomonas fluorescens ATCC 11253 cultivated in a vegetable-based broth. Transmission and scanning electron microscopy images of bacterial cells exposed to CAR and CIN alone or in combination showed marked ultrastructural changes after 1h of exposure. These changes included shrunken protoplasm, discontinuity of the outer and cytoplasmic membranes and leakage of the intracellular material. Confocal scanning laser microscopy images corroborated the electron microscopy data, showing a decrease in the number of SYTO-9 cells (intact cells) with a concomitant increase in the number of PI-positive cells (dead cells). All of these morphological changes are indicative of increased membrane permeability and the loss of bacterial envelope integrity, which ultimately lead to cell death. The combination of sublethal concentrations of CAR and CIN could be applied to inhibit the growth of P. fluorescens on vegetables. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. DETECTION OF PHENOL DEGRADING BACTERIA AND PSEUDOMONAS PUTIDA IN ACTIVATED SLUDGE BY POLYMERASE CHAIN REACTION

    Directory of Open Access Journals (Sweden)

    H. Movahedyan ، H. Khorsandi ، R. Salehi ، M. Nikaeen

    2009-04-01

    Full Text Available Phenol is one of the organic pollutants in various industrial wastewaters especially petrochemical and oil refining. Biological treatment is one of the considerable choices for removing of phenol present in these wastewaters. Identification of effective microbial species is considered as one of the important priorities for production of the biomass in order to achieve desirable kinetic of biological reactions. Basic purpose of this research is identification of phenol-degrading Pseudomonas Putida in activated sludge by polymerase chain reaction (PCR that has high speed and specificity. In this research, 10 various colonies of phenol-degrading bacteria were isolated from municipal activated sludge and the rate of phenol removal and growth rate of these bacteria were assessed in different concentrations of phenol (200 – 900 mg/L. Confirmation of the largest subunit of multicomponent phenol hydroxylase (LmPH gene and gene coding the N fragment in Pseudomonas Putida-derived methyl phenol operon (DmpN gene through PCR were used for general identification of phenol-degrading bacteria and Pseudomonas Putida, respectively. Presence of a 600 bp (base pairs bond in all of isolated strains indicated that they contain phenol hydroxylase gene. 6 of 10 isolated bacteria were Pseudomonas Putida because they produced a 199 bp PCR product by DmpN primers. According to PCR results in this study, the best phenol-degrading bacteria that can utilize 500 – 600 mg/L phenol completely after 48 hours incubation, belong to Pseudomonas Putida strains. It is clear that use of isolated bacteria can lead to considerable decrease of treatment time as well as promotion of phenol removal rate.

  8. 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...... 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. 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...

  9. 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...

  10. 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 ...

  11. Marine Pseudomonas putida: a potential source of antimicrobial substances against antibiotic-resistant bacteria

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    Palloma Rodrigues Marinho

    2009-08-01

    Full Text Available Bacteria isolated from marine sponges found off the coast of Rio de Janeiro, Brazil, were screened for the production of antimicrobial substances. We report a new Pseudomonas putida strain (designated P. putida Mm3 isolated from the sponge Mycale microsigmatosa that produces a powerful antimicrobial substance active against multidrug-resistant bacteria. P. putida Mm3 was identified on the basis of 16S rRNA gene sequencing and phenotypic tests. Molecular typing for Mm3 was performed by RAPD-PCR and comparison of the results to other Pseudomonas strains. Our results contribute to the search for new antimicrobial agents, an important strategy for developing alternative therapies to treat infections caused by multidrug-resistant bacteria.

  12. Marine Pseudomonas putida: a potential source of antimicrobial substances against antibiotic-resistant bacteria.

    Science.gov (United States)

    Marinho, Palloma Rodrigues; Moreira, Ana Paula Barbosa; Pellegrino, Flávia Lúcia Piffano Costa; Muricy, Guilherme; Bastos, Maria do Carmo de Freire; Santos, Kátia Regina Netto dos; Giambiagi-deMarval, Marcia; Laport, Marinella Silva

    2009-08-01

    Bacteria isolated from marine sponges found off the coast of Rio de Janeiro, Brazil, were screened for the production of antimicrobial substances. We report a new Pseudomonas putida strain (designated P. putida Mm3) isolated from the sponge Mycale microsigmatosa that produces a powerful antimicrobial substance active against multidrug-resistant bacteria. P. putida Mm3 was identified on the basis of 16S rRNA gene sequencing and phenotypic tests. Molecular typing for Mm3 was performed by RAPD-PCR and comparison of the results to other Pseudomonas strains. Our results contribute to the search for new antimicrobial agents, an important strategy for developing alternative therapies to treat infections caused by multidrug-resistant bacteria.

  13. 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

    selective feeding flagellate Cercomonas longicauda versus the non-selective feeding nematode Caenorhabditis elegans) influence the abundance of two bacteria that compete for resources in simple model communities. Microcosms consisted of either one gfp-tagged bacterial strain (Pseudomonas fluorescens DSM...

  14. A bacteria-specific 2[4Fe-4S] ferredoxin is essential in Pseudomonas aeruginosa

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    Diallinas George

    2010-10-01

    Full Text Available Abstract Background Ferredoxins are small iron-sulfur proteins belonging to all domains of life. A sub-group binds two [4Fe-4S] clusters with unequal and extremely low values of the reduction potentials. These unusual properties are associated with two specific fragments of sequence. The functional importance of the very low potential ferredoxins is unknown. Results A bioinformatic screening of the sequence features defining very low potential 2[4Fe-4S] ferredoxins has revealed the almost exclusive presence of the corresponding fdx gene in the Proteobacteria phylum, without occurrence in Archaea and Eukaryota. The transcript was found to be monocistronic in Pseudomonas aeruginosa, and not part of an operon in most bacteria. Only fdx genes of bacteria which anaerobically degrade aromatic compounds belong to operons. As this pathway is not present in all bacteria having very low potential 2[4Fe-4S] ferredoxins, these proteins cannot exclusively be reductants of benzoyl CoA reductases. Expression of the ferredoxin gene did not change in response to varying growth conditions, including upon macrophage infection or aerobic growth with 4-hydroxy benzoate as carbon source. However, it increased along the growth curve in Pseudomonas aeruginosa and in Escherichia coli. The sequence immediately 5' upstream of the coding sequence contributed to the promotor activity. Deleting the fdx gene in Pseudomonas aeruginosa abolished growth, unless a plasmid copy of the gene was provided to the deleted strain. Conclusions The gene of the very low potential 2[4Fe-4S] ferredoxin displays characteristics of a housekeeping gene, and it belongs to the minority of genes that are essential in Pseudomonas aeruginosa. These data identify a new potential antimicrobial target in this and other pathogenic Proteobacteria.

  15. PURIFIKASI DAN KARAKTERISASI PROTEASE DARI BAKTERI PATOGEN Pseudomonas aeruginosa [Purification and Characterization of Protease from Pathogenic Bacteria Pseudomonas aeruginosa

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    Ace Baehaki1

    2008-06-01

    Full Text Available In the last decade, concern on protease as medical target for overcoming bacterial diseases and viral diseases has been rapidly increased because of the obvious involvement of this enzyme in the molecular of the diseases. The purpose of this research was to purify and characterize protease from pathogenic bacteria Pseudomonas aeruginosa. The bacteria were grown in media containing triptone 1%, NaCl 1% and Yeast extract 0,5%. Protease of P.aeruginosa was purified using column chromatography with Sephadex G-100 gel. There were three peaks of enzyme protein, which were detected on fractions 14, 17 and 30. The optimum pH of the extracelluler protease from P. aeruginosa was 8. The optimum temperature of P.aeruginosa protease was 300C. Fe3+ (1dan 5 mM was strong activator and Co2+ was strong inhibitor. Study on the effect of metals ion and spesific inhibitors indicated that protease from P. aeruginosa was serin metaloprotease. The apparent moleculer weights, as determined by SDS-PAGE and zymogram technique, 36 kD and 42 kD.

  16. Alkaline lipase from Pseudomonas fluorescens non-covalently immobilised on pristine versus oxidised multi-wall carbon nanotubes as efficient and recyclable catalytic systems in the synthesis of Solketal esters.

    Science.gov (United States)

    Boncel, Sławomir; Zniszczoł, Aurelia; Szymańska, Katarzyna; Mrowiec-Białoń, Julita; Jarzębski, Andrzej; Walczak, Krzysztof Z

    2013-09-10

    In order to produce effective and recyclable catalysts for enantioselective transesterification in the industrial applications, alkaline lipase from Pseudomonas fluorescens was non-covalently immobilised (ca. 6 wt%) on pristine multi-wall carbon nanotubes (MWCNTs) and oxidised MWCNTs (O-MWCNTs) using an adsorption technique. Mesoporous silica modified with n-octyl groups was used as a reference support. Irreversible transesterifications of three vinyl esters (acyl donors) by racemic Solketal with a chromatographically (GC) traced kinetics were selected as model reactions. The undertaken comparative studies revealed that different morphology and chemical functionalisation of the supports led to various enzyme loadings, catalytic activities and enantioselectivities. MWCNT-lipase emerged as the exceptionally active (yield up to 98%, t=1h, 1320 Ug(-1), i.e. 9 times more active than native enzyme) whereas lipase immobilised on O-MWCNTs as the most enantioselective system, particularly for longer acyl chain esters (e.e. up to 72% after 30 min at yield of 20%, 340 Ug(-1)). Moreover, the activity of all nanotube-based catalysts after 10 cycles of transesterification remained practically unchanged. The differences in performance of MWCNTs and O-MWCNTs as solid supports were found to be based on geometry of pores, dominating hydrophobic interactions and absence/presence of the surface polar groups. Due to an excellent activity and reusability of the nanotube-lipase catalysts one can propose (O-)MWCNT as supports of a prospective industrial relevance. Copyright © 2013 Elsevier Inc. All rights reserved.

  17. Metabolic engineering of bacteria for environmental applications: construction of Pseudomonas strains for biodegradation of 2-chlorotoluene.

    Science.gov (United States)

    Haro, M A; de Lorenzo, V

    2001-02-13

    In this article, we illustrate the challenges and bottlenecks in the metabolic engineering of bacteria destined for environmental bioremediation, by reporting current efforts to construct Pseudomonas strains genetically designed for degradation of the recalcitrant compound 2-chlorotoluene. The assembled pathway includes one catabolic segment encoding the toluene dioxygenase of the TOD system of Pseudomonas putida F1 (todC1C2BA), which affords the bioconversion of 2-chlorotoluene into 2-chlorobenzaldehyde by virtue of its residual methyl-monooxygenase activity on o-substituted substrates. A second catabolic segment encoded the entire upper TOL pathway from pWW0 plasmid of P. putida mt-2. The enzymes, benzyl alcohol dehydrogenase (encoded by xylB) and benzaldehyde dehydrogenase (xylC) of this segment accept o-chloro-substituted substrates all the way down to 2-chlorobenzoate. These TOL and TOD segments were assembled in separate mini-Tn5 transposon vectors, such that expression of the encoded genes was dependent on the toluene-responsive Pu promoter of the TOL plasmid and the cognate XylR regulator. Such gene cassettes (mini-Tn5 [UPP2] and mini-Tn5 [TOD2]) were inserted in the chromosome of the 2-chlorobenzoate degraders Pseudomonas aeruginosa PA142 and P. aeruginosa JB2. GC-MS analysis of the metabolic intermediates present in the culture media of the resulting strains verified that these possessed, not only the genetic information, but also the functional ability to mineralise 2-chlorotoluene. However, although these strains did convert the substrate into 2-chlorobenzoate, they failed to grow on 2-chlorotoluene as the only carbon source. These results pinpoint the rate of the metabolic fluxes, the non-productive spill of side-metabolites and the physiological control of degradative pathways as the real bottlenecks for degradation of certain pollutants, rather than the theoretical enzymatic and genetic fitness of the recombinant bacteria to the process. Choices to

  18. 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.

  19. High mannose-binding antiviral lectin PFL from Pseudomonas fluorescens Pf0-1 promotes cell death of gastric cancer cell MKN28 via interaction with α2-integrin.

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    Yuichiro Sato

    Full Text Available Novel anti-HIV lectin family which shows a strict binding specificity for high mannose glycans has been found in lower organisms. The bacterial orthologue has been identified in the genome of Pseudomonas fluorescens Pf0-1 and the gene coding a putative lectin was cloned, expressed in Escherichia coli and purified by one step gel filtration. Glycan array screening of the recombinant lectin, termed PFL, has revealed that PFL preferentially recognizes high mannose glycans with α1-3 Man that was highly exposed at the D2 position. In contrast, masking of this α1-3 Man with α1-2 Man dramatically impaired lectin-carbohydrate interactions. Reducing terminal disaccharide, GlcNAc-GlcNAc of high mannose glycans was also essential for PFL-binding. PFL showed a potent anti-influenza virus activity by inhibiting the virus entry into cells at doses of low nanomolar concentration. At micromolar concentration or higher, PFL showed a cytotoxicity accompanying loss of the cell adhesion against human gastric cancer MKN28 cells. The cell surface molecule to which PFL bound was co-precipitated with biotin-labeled PFL and identified as integrin α2 by peptide mass fingerprinting using MALDI-TOF mass spectrometry. Intriguingly, upon treatment with exogenous PFL, integrin α2 on the cell surface underwent rapid internalization to the cytoplasm and accumulated to perinuclear region, together with the bound PFL. The resulting loss of cell adherence would trigger a signaling pathway that induced anoikis-like cell death. These events were effectively inhibited by pretreatment of PFL with mannnan, indicating the involvement of high mannose glycans on PFL-induced cell death that was triggered by PFL-integrin α2 interactions.

  20. Evaluation of economically feasible, natural plant extract-based microbiological media for producing biomass of the dry rot biocontrol strain Pseudomonas fluorescens P22Y05 in liquid culture.

    Science.gov (United States)

    Khalil, Sadia; Ali, Tasneem Adam; Skory, Chris; Slininger, Patricia J; Schisler, David A

    2016-02-01

    The production of microbial biomass in liquid media often represents an indispensable step in the research and development of bacterial and fungal strains. Costs of commercially prepared nutrient media or purified media components, however, can represent a significant hurdle to conducting research in locations where obtaining these products is difficult. A less expensive option for providing components essential to microbial growth in liquid culture is the use of extracts of fresh or dried plant products obtained by using hot water extraction techniques. A total of 13 plant extract-based media were prepared from a variety of plant fruits, pods or seeds of plant species including Allium cepa (red onion bulb), Phaseolus vulgaris (green bean pods), and Lens culinaris (lentil seeds). In shake flask tests, cell production by potato dry rot antagonist Pseudomonas fluorescens P22Y05 in plant extract-based media was generally statistically indistinguishable from that in commercially produced tryptic soy broth and nutrient broth as measured by optical density and colony forming units/ml produced (P ≤ 0.05, Fisher's protected LSD). The efficacy of biomass produced in the best plant extract-based media or commercial media was equivalent in reducing Fusarium dry rot by 50-96% compared to controls. In studies using a high-throughput microbioreactor, logarithmic growth of P22Y05 in plant extract-based media initiated in 3-5 h in most cases but specific growth rate and the time of maximum OD varied as did the maximum pH obtained in media. Nutrient analysis of selected media before and after cell growth indicated that nitrogen in the form of NH4 accumulated in culture supernatants, possibly due to unbalanced growth conditions brought on by a scarcity of simple sugars in the media tested. The potential of plant extract-based media to economically produce biomass of microbes active in reducing plant disease is considerable and deserves further research.

  1. Soil burial method for plastic degradation performed by Pseudomonas PL-01, Bacillus PL-01, and indigenous bacteria

    Science.gov (United States)

    Shovitri, Maya; Nafi'ah, Risyatun; Antika, Titi Rindi; Alami, Nur Hidayatul; Kuswytasari, N. D.; Zulaikha, Enny

    2017-06-01

    Lately, plastic bag is becoming the most important pollutant for environment since it is difficult to be naturally degraded due to it consists of long hydrocarbon polymer chains. Our previous study indicated that our pure isolate Pseudomonas PL-01 and Bacillus PL-01 could degrade about 10% plastic bag. This present study was aimed to find out whether Pseudomonas PL01 and Bacillus PL01 put a positive effect to indigenous bacteria from marginal area in doing plastic degradation with a soil burial method. Beach sand was used as a representative marginal area, and mangrove sediment was used as a comparison. Plastics were submerged into unsterile beach sand with 10% of Pseudomonas PL-01 or Bacillus PL-01 containing liquid minimal salt medium (MSM) separately, while other plastics were submerged into unsterile mangrove sediments. After 4, 8, 12 and 16 weeks, their biofilm formation on their plastic surfaces and plastic degradation were measured. Results indicated that those 2 isolates put positive influent on biofilm formation and plastic degradation for indigenous beach sand bacteria. Bacillus PL-01 put higher influent than Pseudomonas PL-01. Plastic transparent was preferable degraded than black and white plastic bag `kresek'. But anyhow, indigenous mangrove soil bacteria showed the best performance in biofilm formation and plastic degradation, even without Pseudomonas PL-01 or Bacillus PL-01 addition. Fourier Transform Infrared (FTIR) analysis complemented the results; there were attenuated peaks with decreasing peaks transmittances. This FTIR peaks indicated chemical functional group changes happened among the plastic compounds after 16 weeks incubation time.

  2. 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.

  3. Anti-Biofilm Activities from Marine Cold Adapted Bacteria Against Staphylococci and Pseudomonas aeruginosa.

    Science.gov (United States)

    Papa, Rosanna; Selan, Laura; Parrilli, Ermenegilda; Tilotta, Marco; Sannino, Filomena; Feller, Georges; Tutino, Maria L; Artini, Marco

    2015-01-01

    Microbial biofilms have great negative impacts on the world's economy and pose serious problems to industry, public health and medicine. The interest in the development of new approaches for the prevention and treatment of bacterial adhesion and biofilm formation has increased. Since, bacterial pathogens living in biofilm induce persistent chronic infections due to the resistance to antibiotics and host immune system. A viable approach should target adhesive properties without affecting bacterial vitality in order to avoid the appearance of resistant mutants. Many bacteria secrete anti-biofilm molecules that function in regulating biofilm architecture or mediating the release of cells from it during the dispersal stage of biofilm life cycle. Cold-adapted marine bacteria represent an untapped reservoir of biodiversity able to synthesize a broad range of bioactive compounds, including anti-biofilm molecules. The anti-biofilm activity of cell-free supernatants derived from sessile and planktonic cultures of cold-adapted bacteria belonging to Pseudoalteromonas, Psychrobacter, and Psychromonas species were tested against Staphylococcus aureus, Staphylococcus epidermidis, and Pseudomonas aeruginosa strains. Reported results demonstrate that we have selected supernatants, from cold-adapted marine bacteria, containing non-biocidal agents able to destabilize biofilm matrix of all tested pathogens without killing cells. A preliminary physico-chemical characterization of supernatants was also performed, and these analyses highlighted the presence of molecules of different nature that act by inhibiting biofilm formation. Some of them are also able to impair the initial attachment of the bacterial cells to the surface, thus likely containing molecules acting as anti-biofilm surfactant molecules. The described ability of cold-adapted bacteria to produce effective anti-biofilm molecules paves the way to further characterization of the most promising molecules and to test their

  4. Anti-biofilm activities from marine cold adapted bacteria against staphylococci and Pseudomonas aeruginosa

    Directory of Open Access Journals (Sweden)

    Rosanna ePapa

    2015-12-01

    Full Text Available Microbial biofilms have great negative impacts on the world’s economy and pose serious problems to industry, public health and medicine. The interest in the development of new approaches for the prevention and treatment of bacterial adhesion and biofilm formation has increased. Since, bacterial pathogens living in biofilm induce persistent chronic infections due to the resistance to antibiotics and host immune system. A viable approach should target adhesive properties without affecting bacterial vitality in order to avoid the appearance of resistant mutants. Many bacteria secrete anti-biofilm molecules that function in regulating biofilm architecture or mediating the release of cells from it during the dispersal stage of biofilm life cycle. Cold-adapted marine bacteria represent an untapped reservoir of biodiversity able to synthesize a broad range of bioactive compounds, including anti-biofilm molecules.The anti-biofilm activity of cell-free supernatants derived from sessile and planktonic cultures of cold-adapted bacteria belonging to Pseudoalteromonas, Psychrobacter and Psychromonas species were tested against Staphylococcus aureus, Staphylococcus epidermidis and Pseudomonas aeruginosa strains. Reported results demonstrate that we have selected supernatants, from cold-adapted marine bacteria, containing non-biocidal agents able to destabilize biofilm matrix of all tested pathogens without killing cells. A preliminary physico-chemical characterization of supernatants was also performed, and these analyses highlighted the presence of molecules of different nature that act by inhibiting biofilm formation. Some of them are also able to impair the initial attachment of the bacterial cells to the surface, thus likely containing molecules acting as anti-biofilm surfactant molecules.The described ability of cold-adapted bacteria to produce effective anti-biofilm molecules paves the way to further characterization of the most promising molecules

  5. Excretions/secretions from bacteria-pretreated maggot are more effective against Pseudomonas aeruginosa biofilms.

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    Ke-chun Jiang

    Full Text Available BACKGROUND: Sterile larvae--maggots of the green bottle blowfly Lucilia sericata are employed as a treatment tool for various types of chronic wounds. Previous studies reported that excretions/secretions (ES of the sterile larvae could prevent and remove the biofilms of various species of bacteria. In the present study we assessed the effect of ES from the larvae pretreated with Pseudomonas aeruginosa on the bacteria biofilms. METHODS AND FINDINGS: We investigated the effects of ES from the maggot pretreated with P. aeruginosa on the biofilms using microtitre plate assays and on bactericidal effect using the colony-forming unit (CFU assay. The results showed that only 30 µg of the ES from the pretreated maggots could prevent and degrade the biofilm of P. aeruginosa. However, the CFU count of P. aeruginosa was not decrease when compared to the ES from non pretreated maggots in this study condition. It is suggested that the ES from the pretreated maggot was more effective against biofilm of P. aeruginosa than sterile maggot ES. CONCLUSIONS: Our results showed that the maggot ES, especially the bacteria-pretreated larva ES may provide a new insight into the treatment tool of the bacterial biofilms.

  6. Surveillance of bacteria Pseudomonas aeruginosa and MRSA associated with chronic suppurative otitis media

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    Sibanarayan Rath

    Full Text Available Abstract Introduction: Suppurative otitis media is a critical disease causing perforation of the tympanic membrane associated with changes of the mucoperiosteum of the middle ear cleft. Objective: To isolate causative bacteria from chronic suppurative ear discharges and to ascertain their antibiotic profiles, of patients attending outpatients department in 3 years. Methods: For isolation of bacteria, samples of ear discharges were grown in suitable media and bacteria were subjected to antibiotic profiling by the Kirby-Bauer's method with presently used antibiotics. Results: A total of 1043 bacteria were isolated, including Pseudomonas aeruginosa and methicillin resistant Staphylococcus aureus, along with 121 fungal isolates. Among 371 P. aeruginosa isolates, tobramycin 30 had the highest susceptibility rate 93.2%, followed by ceftazidime 30, 91.5% and amikacin 10 µg/disk 64.4%. Of 359 S. aureus isolates, there were 236 coagulase negative S. aureus + methicillin sensitive S. aureus isolates, while 123 isolates were methicillin resistant Staphylococcus aureus with 95.2% isolates susceptible to cloxacillin 15, 83.3% isolates to erythromycin 15 and 78.5% isolates to gentamicin 30 µg/disk. Of 1164, 49 patients presented post aural abscess, 12 patients had intracranial complications, 9 patients had facial palsy and 3 patients had labyrinthitis. More than 90% P. aeruginosa and 90% S. aureus isolates were sensitive to tobramycin 30 and cloxacillin 30 µg/disk, respectively. Conclusion: Multidrug resistant strains of P. aeruginosa were more prevalent than those of S. aureus in ear discharges. Tobramycin and cloxacillin may be included in the formulatory antibiotic regimen to overcome bacterial infections in chronic suppurative otitis media.

  7. Dissolution of Al-substituted goethites by an aerobic Pseudomonas mendocina var. bacteria

    Science.gov (United States)

    Maurice, P. A.; Lee, Y.-J.; Hersman, L. E.

    2000-04-01

    Goethite particles in soil environments often contain Al 3+ substituted for Fe 3+ in octahedrally coordinated sites. Al substitution has been shown to alter mineral stability and abiotic dissolution rates. This study focused on the effects of Al substitution (to 8.8 mol%) on synthetic goethite dissolution by an aerobic Pseudomonas mendocina var. bacteria. In contrast to dissimilatory iron reducing bacteria (DIRB), this bacteria is not capable of using Fe as a terminal electron acceptor for oxidative phosphorylation, and hence only requires μM concentrations of Fe for metabolism. Pure and substituted goethites were reacted with microorganisms in Fe-limited growth media wherein the only source of Fe was the solid phase, so that microbial populations could only grow by obtaining Fe through mineral dissolution. Because at least some Fe was taken up by the bacteria, we could not measure Fe release rates directly from dissolved Fe concentrations. Rather, we relied upon microbial growth measurements as indirect indicators of mineral dissolution. Increasing Al substitution resulted in particles with progressively decreasing mean particle length and aspect ratios, as well as fewer domains, as measured by atomic-force microscopy (AFM); but with increasing structural order as determined by XRD line widths. Experiments conducted in the dark at 22°C, exposed to the atmosphere, showed that maximum microbial population did not correlate with particle specific surface area, which is in contrast with previous studies using DIRB. Maximum microbial population increased a small amount with increasing Al content of the goethites, in contrast with several previous investigations of abiotic dissolution. Because dense biofilms formed, we were unable to use AFM to observe mineral dissolution features. AFM imaging suggested that more highly substituted goethites formed denser aggregates, and previous investigations have shown that aggregate structure is important for microbial attachment

  8. Transesterification of Jatropha oil using immobilized Pseudomonas ...

    African Journals Online (AJOL)

    mild transesterification has become of much current interest for alternative fuel production. In the present study the ability of a commercial immobilized Pseudomonas fluorescens MTCC 103 to catalyze the transesterification of Jatropha oil and methanol was investigated. The cell of P. fluorescens was easily immobilized ...

  9. Carbazole-degradative IncP-7 plasmid pCAR1.2 is structurally unstable in Pseudomonas fluorescens Pf0-1, which accumulates catechol, the intermediate of the carbazole degradation pathway.

    Science.gov (United States)

    Takahashi, Yurika; Shintani, Masaki; Li, Li; Yamane, Hisakazu; Nojiri, Hideaki

    2009-06-01

    We determined the effect of the host on the function and structure of the nearly identical IncP-7 carbazole-degradative plasmids pCAR1.1 and pCAR1.2. We constructed Pseudomonas aeruginosa PAO1(pCAR1.2) and P. fluorescens Pf0-1Km(pCAR1.2) and compared their growth on carbazole- and succinate-containing media with that of P. putida KT2440(pCAR1.1). We also assessed the stability of the genetic structures of the plasmids in each of the three hosts. Pf0-1Km(pCAR1.2) showed dramatically delayed growth when carbazole was supplied as the sole carbon source, while the three strains grew at nearly the same rate on succinate. Among the carbazole-grown Pf0-1Km(pCAR1.2) cells, two types of deficient strains appeared and dominated the population; such dominance was not observed in the other two strains or for succinate-grown Pf0-1Km(pCAR1.2). Genetic analysis showed that the two deficient strains possessed pCAR1.2 derivatives in which the carbazole-degradative car operon was deleted or its regulatory gene, antR, was deleted by homologous recombination between insertion sequences. From genomic information and quantitative reverse transcription-PCR analyses of the genes involved in carbazole mineralization by Pf0-1Km(pCAR1.2), we found that the cat genes on the chromosome of Pf0-1Km, which are necessary for the degradation of catechol (a toxic intermediate in the carbazole catabolic pathway), were not induced in the presence of carbazole. The resulting accumulation of catechol may have enabled the strain that lost its carbazole-degrading ability to have overall higher fitness than the wild-type strain. These results suggest that the functions of the chromosomal genes contributed to the selection of plasmid derivatives with altered structures.

  10. RND efflux pump mediated antibiotic resistance in Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa: a major issue worldwide.

    Science.gov (United States)

    Puzari, Minakshi; Chetia, Pankaj

    2017-02-01

    Therapeutic failures against diseases due to resistant Gram-negative bacteria have become a major threat nowadays as confirmed by surveillance reports across the world. One of the methods of development of multidrug resistance in Escherichia coli and Pseudomonas aeruginosa is by means of RND efflux pumps. Inhibition of these pumps might help to combat the antibiotic resistance problem, for which the structure and regulation of the pumps have to be known. Moreover, judicious antibiotic use is needed to control the situation. This paper focuses on the issue of antibiotic resistance as well as the structure, regulation and inhibition of the efflux pumps present in Escherichia coli and Pseudomonas aeruginosa.

  11. Does the presence of bacteria effect basaltic glass dissolution rates? 1: Dead Pseudomonas reactants

    Science.gov (United States)

    Stockmann, Gabrielle J.; Shirokova, Liudmila S.; Pokrovsky, Oleg S.; Oelkers, Eric H.; Benezeth, Pascale

    2010-05-01

    Basaltic glass and crystalline basalt formations in Iceland have been suggested for industrial CO2 storage due to their porous and permeable properties and high reactivity. Acid CO2-saturated waters in contact with basaltic glass will lead to rapid dissolution of the glass and release of divalent cations, (Ca2+, Mg2+, Fe2+) that can react to form stable carbonates and thereby trap the CO2. However, the basalt formations in Iceland not only contains glass and mineral assemblages, but also host microbiological communities that either by their presence or by active involvement in chemical reactions could affect the amount of basaltic glass being dissolved and CO2 being trapped. Samples of natural bacteria communities from the CO2 storage grounds in Iceland were collected, separated, and purified using agar plate technique and cultured under laboratory conditions in nutrient broth-rich media. Heterotrophic aerobic Gram-negative strain of Pseudomonas reactants was selected for a series of flow-through experiments aimed at evaluation of basaltic glass dissolution rate in the presense of increasing amounts of dead bacteria and their lysis products. The experiments were carried out using mixed-flow reactors at pH 4, 6, 8 and 10 at 25 °C. Each of the four reactors contained 1 gram of basaltic glass of the size fraction 45-125 μm. This glass was dissolved in ~ 0.01 M buffer solutions (acetate, MES, bicarbonate and carbonate+bicarbonate mixture) of the desired pH. All experiments ran 2 months, keeping the flowrate and temperature stable and only changing the concentration of dead bacteria in the inlet solutions (from 0 to 430 mg/L). Experiments were performed in sterile conditions, and bacterial growth was prevented by adding NaN3 to the inlet solutions. Routine culturing of bacteria on the agar plates confirmed the sterility of experiments. Samples of outlet solutions were analyzed for major cations and trace elements by ICP-MS. Results demonstrate a slight decrease in the

  12. Chemical resistance of the gram-negative bacteria to different sanitizers in a water purification system

    Directory of Open Access Journals (Sweden)

    Penna Thereza CV

    2006-08-01

    Full Text Available Abstract Background Purified water for pharmaceutical purposes must be free of microbial contamination and pyrogens. Even with the additional sanitary and disinfecting treatments applied to the system (sequential operational stages, Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas alcaligenes, Pseudomonas picketti, Flavobacterium aureum, Acinetobacter lowffi and Pseudomonas diminuta were isolated and identified from a thirteen-stage purification system. To evaluate the efficacy of the chemical agents used in the disinfecting process along with those used to adjust chemical characteristics of the system, over the identified bacteria, the kinetic parameter of killing time (D-value necessary to inactivate 90% of the initial bioburden (decimal reduction time was experimentally determined. Methods Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas alcaligenes, Pseudomonas picketti, Flavobacterium aureum, Acinetobacter lowffi and Pseudomonas diminuta were called in house (wild bacteria. Pseudomonas diminuta ATCC 11568, Pseudomonas alcaligenes INCQS , Pseudomonas aeruginosa ATCC 15442, Pseudomonas fluorescens ATCC 3178, Pseudomonas picketti ATCC 5031, Bacillus subtilis ATCC 937 and Escherichia coli ATCC 25922 were used as 'standard' bacteria to evaluate resistance at 25°C against either 0.5% citric acid, 0.5% hydrochloric acid, 70% ethanol, 0.5% sodium bisulfite, 0.4% sodium hydroxide, 0.5% sodium hypochlorite, or a mixture of 2.2% hydrogen peroxide (H2O2 and 0.45% peracetic acid. Results The efficacy of the sanitizers varied with concentration and contact time to reduce decimal logarithmic (log10 population (n cycles. To kill 90% of the initial population (or one log10 cycle, the necessary time (D-value was for P. aeruginosa into: (i 0.5% citric acid, D = 3.8 min; (ii 0.5% hydrochloric acid, D = 6.9 min; (iii 70% ethanol, D = 9.7 min; (iv 0.5% sodium bisulfite, D = 5.3 min; (v 0.4% sodium hydroxide, D = 14.2 min; (vi 0.5% sodium

  13. Glyphosate biodegradation by plant growth promoting bacteria and their effect to paddy germination in glyphosate contaminated soil

    Directory of Open Access Journals (Sweden)

    Lutfi Tri Andriani

    2017-10-01

    Full Text Available Glyphosate is the most widely used herbicide in Indonesia. Glyphosate persistence between 55 days to 3 years. Widespread and uncontrolled use can cause weeds to become resistant and residue contaminates the soil and water environment. Due to the residual impact of glyphosate, it is necessary to identify a method that can increase the degradation of glyphosate. Several studies have shown that glyphosate can be degraded by microorganisms (fungi, rhizosphere and endophytic bacteria, some of which are members of plant growth-promoting bacteria. This study used the bacteria Enterobacter cloacae, Enterobacter sp and Pseudomonas fluorescens. These three types of bacteria have growth-promoting properties and potentially increase glyphosate degradation. Results of chromatogram on the residual test of glyphosate in liquid medium and soil containing glyphosate showed that glyphosate residue decreased with the addition of bacterial treatment when compared to control. The percentage of degradation in liquid medium are 96.06%  by Enterobacter cloacae, 57% by Enterobacter spand 93.45%  by Pseudomonas fluorescens.The percentage of degradation in soil medium are 4.32%  by Enterobacter cloacae, 23.49% by Enterobacter spand 12.19% by Pseudomonas fluorescens.A positive result indicates that bacterial growth boosters from the plant (endophyte as well as the area of rooting (rhizosphere have additional potential as biofertilizer, bio stimulant, bio protectant but also as bio degradator pollutants such as the herbicide glyphosate

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

    African Journals Online (AJOL)

    Michael Horsfall

    (Potassium sodium tartarate) and 1% (w/v) sodium hydroxide. To prepare 1 – litre of solution, 10 g of sodium hydroxide pellet was dissolved in 1- litre of distilled water. To this was added 10 g of powdered. 3,5 – Di-nitrosalicylic acid, 0.5 g of sodium bisulphite, 2.52 ml of 80% Phenol and stirred to complete dissolution.

  15. Characterization and identification of Pseudomonas fluorescens ...

    African Journals Online (AJOL)

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  16. Effect of Pseudomonas and Bacillus bacteria on Yield and Nutrient Uptake in Comparison with Chemical and Organic Fertilizers in Wheat

    Directory of Open Access Journals (Sweden)

    A. Fallah Nosrat Abad

    2015-06-01

    Full Text Available The high cost of fertilizers in farming systems, soil pollution and degradation of soil are factors that caused to full use of available renewable nutrient sources of plant (organic and biological with optimal application of fertilizers in order to maintain fertility, structure, biological activity, exchange capacity and water-holding capacity of the water in soil. Therefore, in recent years, according to investigators biofertilizers and organic farming as an alternative to chemical fertilizers has been drawn. Through this study, we examined the effects of triple superphosphate, organic matters and phosphate solubilizing microorganisms on quantitative and qualitative yield of wheat and nutrient uptake. The experiment was carried out in the factorial based on randomized complete block design. The factors were: 1-phosphate solubilizing bacteria in three levels including control, Pseudomonas Putida and Bacillus Coagulans bacteria, 2- triple superphosphate in five levels of 0, 25%, 50%, 75% and 100% and 3-organic matter in 2 levels of 0 and 15 ton/ha in the soil with high phosphorous accessibility (13 mg/kg soil but lower than sufficient limit for plant 15 mg/kg soil. The results showed that the highest amount of yield has been recorded in Pseudomonas Putida bacteria treatment with organic matter and 25% phosphate fertilizer. As a result, at the conditions of this experiment phosphate solubilizing bacteria and organic matter significantly had higher yield than control and their combination with phosphate fertilizer had significant effect on reducing phosphate fertilizer use.

  17. The Survey of Withani somnifera Extraction against Resistant Strains of Pseudomonas aeruginosa Bacteria to Selective Antibiotics

    Directory of Open Access Journals (Sweden)

    Mohammad Bokaeian

    2015-11-01

    Full Text Available Introduction:  Due  to  more  resistance  of  pathogenic  bacteria  to  new  and  current antibiotics  researchers  are  looking  to  find  the  agents  of  herbal  with  antimicrobial activities in order to replace chemical drugs.Methods:   The herbal extract of Withani somnifera was done by using a rotary vacuum,20 strains of Pseudomons aeruginosa were isolated from urinary infections hospitalized patients  in  city of Zabol  hospital.  The  MIC  Withani  somnifera  were  determined  by dilution method in various concentrations. Sensitivity of strains to multiple antibiotics was evaluated by standard disk diffusion Kirby-Bauer.Results:    The  result  showed  that  P.  aeruginosa  were  resistance  to  4  of the  agents including ampicillin  (85%, nitrofurantoin  (65%, nalidixic acid  (65%, ciprofloxacin (15% and for 5 strains of Pseudomonas showed MIC with activity of 100 ppm.Conclusion:   This  study  has  suggested  the  effect  of  winter  cherry  extract  on  P. aeruginosa in the in vitro assay. It s effectiveness of on in vivo system can be examined in future.

  18. Regulation of synthesis of key naphthalene catabolism enzymes in Pseudomonas putida and Pseudomonas fluorescens carrying biodegradation plasmids NAH, pBS3, pBS2 and NPL-1

    Energy Technology Data Exchange (ETDEWEB)

    Starovoytov, I.I.

    A study was made of the regulation of catabolism of naphthalene in pseudomonada containing several biodegradation plasmids. Bacteria were grown on a minimal medium. Induction experiments were performed on a circular rocking device in 750 ml flasks containing 100 ml of medium. Salicylic acid is found to induce key naphthalene catabolism enzymes controlled by the NAH plasmid. Of the key enzymes controlled by NAH plasmid, the synthesis of naphthalene oxygen as salicylate hydroxylase is regulated in a coordinated manner, indicating that these enzymes are included in a single regulatory unit. The presence of proportionality in changes in specific activities of salicylate dehydroxylase and KO2, 3 with induction by salicylate, anthranylate and 3-methylsalicylate indicate that the synthesis of these enzymes is regulated in coordination. The differences in coordination of regulation of the synthesis of naphthalene catabolism enzymes in microorganisms belonging to the same genus and even species probably reflects an independent evolution of the genetic material. The genetic environment of the host cell probably also influences expression of the plasmids. 15 references, 5 figures.

  19. Initial Host Response to Bacteria in the Murine Lung Differs Between Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus pneumoniae.

    Science.gov (United States)

    Preu, Liselotte; Bischoff, Markus; Veith, Nils T; Rosenbruch, Martin; Theegarten, Dirk; Laschke, Matthias W; Meier, Carola; Tschernig, Thomas

    2016-04-01

    Phagocytosis of bacteria is an important process during early host defence. It has been directly observed only ex vivo or in vitro. Here, we report on the observation of phagocytosis under in vivo conditions by using intravital microscopy in the murine lung. Suspensions of fluorescently labelled Streptococcus pneumoniae, Staphylococcus aureus and Pseudomonas aeruginosa cells were each instilled intratracheally to anaesthetized mice. After thoracotomy, the alveolar surface was observed for 30 min. Alveolar phagocytes exhibiting ingested bacteria could be detected and counted. The highest numbers were found after the infection with P. aeruginosa. By using intravital microscopy, cellular host defence could be observed in living mice lungs. The initial phagocytic reaction crucially depends on the species of applied bacteria invading the lung.

  20. Identification and discrimination of Pseudomonas aeruginosa bacteria grown in blood and bile by laser-induced breakdown spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Rehse, Steven J. [Department of Physics and Astronomy, Wayne State University, Detroit, MI 48201 (United States)], E-mail: rehse@wayne.edu; Diedrich, Jonathan [Department of Physics and Astronomy, Wayne State University, Detroit, MI 48201 (United States)], E-mail: jon.diedrich@gd-ais.com; Palchaudhuri, Sunil [Department of Immunology and Microbiology, Wayne State University, Detroit, MI 48201 (United States)], E-mail: spalchau@med.wayne.edu

    2007-10-15

    Pseudomonas aeruginosa bacteria colonies have been analyzed by laser-induced breakdown spectroscopy using nanosecond laser pulses. LIBS spectra were obtained after transferring the bacteria from a nutrient-rich culture medium to a nutrient-free agar plate for laser ablation. To study the dependence of the LIBS spectrum on growth and environmental conditions, colonies were cultured on three different nutrient media: a trypticase soy agar (TSA) plate, a blood agar plate, and a medium chosen deliberately to induce bacteria membrane changes, a MacConkey agar plate containing bile salts. Nineteen atomic and ionic emission lines in the LIBS spectrum, which was dominated by inorganic elements such as calcium, magnesium and sodium, were used to identify and classify the bacteria. A discriminant function analysis was used to discriminate between the P. aeruginosa bacteria and two strains of E. coli: a non-pathogenic environmental strain and the pathogenic strain enterohemorrhagic E. coli 0157:H7 (EHEC). Nearly identical spectra were obtained from P. aeruginosa grown on the TSA plate and the blood agar plate, while the bacteria grown on the MacConkey plate exhibited easily distinguishable differences from the other two. All P. aeruginosa samples, independent of initial growth conditions, were readily discriminated from the two E. coli strains.

  1. Growth ability of Gram negative bacteria in free-living amoebae.

    Science.gov (United States)

    Zeybek, Zuhal; Binay, Ali Rıza

    2014-11-01

    When bacteria and free-living amoebae (FLAs) live both in natural waters and man-made aquatic systems, they constantly interact with each other. Some bacteria can survive and grow within FLAs. Therefore, it has recently been thought that FLAs play an important role in spreading pathogenic bacteria in aquatic systems. In this study we investigated the intracellular growing ability of 7 different Gram-negative bacteria (Pseudomonas fluorescens, Pseudomonas putida, Pasteurella pneumotropica, Aeromonas salmonicida, Legionella pneumophila serogroup 1, L. pneumophila serogroup 3, L. pneumophila serogroup 6) in four different FLA isolates (A1-A4). Among these, four bacterial isolates (P. fluorescens, P.putida, P.pneumotropica, A.salmonicida) and two free-living amoebae isolates (A3, A4) were isolated from the tap water in our city (Istanbul). It was found that 4 different Gram-negative bacteria could grow in A1, 2 different Gram-negative bacteria could grow in A2, 4 different Gram-negative bacteria could grow in A3, 1 Gram-negative bacterium could grow in A4. In conclusion, we think that this ability of growth could vary according to the characteristics of both bacteria and FLA isolates. Also, other factors such as environmental temperature, bacterial concentration, and extended incubation period may play a role in these interactions. This situation can be clarified with future studies. Copyright © 2014 Elsevier Inc. All rights reserved.

  2. Inhibition of quorum sensing in Pseudomonas aeruginosa biofilm bacteria by a halogenated furanone compound

    DEFF Research Database (Denmark)

    Hentzer, Morten; Riedel, Kathrin; Rasmussen, Thomas B

    2002-01-01

    Novel molecular tools have been constructed which allow for in situ detection of N-acyl homoserine lactone (AHL)-mediated quorum sensing in Pseudomonas aeruginosa biofilms. The reporter responds to AHL activation of LasR by expression of an unstable version of the green-fluorescent protein (Gfp)....

  3. Whole cell fatty acid analysis as a tool for classification of phytopathogenic pseudomonas bacteria

    NARCIS (Netherlands)

    Janse, J.D.

    1992-01-01

    In this thesis some members of the plant pathogenic bacterial genus Pseudomonas have been studied. Conventional morphological, biochemical, physiological and pathogenitcity tests as well as a 'finger-print' technique, viz. automated whole cell fatty acid analysis, were

  4. Psychrotrophic bacteria in milk: How much do we really know?

    Directory of Open Access Journals (Sweden)

    Gislene B. de Oliveira

    2015-06-01

    Full Text Available The occurrence of psychrotrophic bacteria in raw milk is studied worldwide due to the difficulties associated with controlling their growth during cold storage and the consequent negative effects upon fluid milk or dairy products. Among the psychrotrophic bacteria, the genus Pseudomonas (represented primarily by P. fluorescens has been highlighted as the cause of numerous defects in dairy products. In light of its perceived predominance, this species has frequently been chosen as a model organism to assess the effects of psychrotrophic bacteria on milk or to evaluate the efficacy of control measures. However, recent findings derived from the application of molecular biological techniques have exposed a number of deficiencies in our knowledge of the biology of milk-associated psychrotrophs. Furthermore, it has been revealed that microbe to microbe communication plays a significant role in determining both the identities and the extent to which different groups of microbes develop during cold storage. The application of molecular identification methods has exposed errors in the classification of members of the genus Pseudomonas isolated from cold stored milk and has stimulated a reevaluation of the presumed status of P. fluorescens as the predominant milk-associated psychrotrophic species. This article presents a succinct review of data from studies on psychrotrophic bacteria in milk, some of which contest established theories in relation to the microbiology of cold stored raw milk, and poses the question: how much do we really know?

  5. Psychrotrophic bacteria in milk: How much do we really know?

    Science.gov (United States)

    de Oliveira, Gislene B; Favarin, Luciana; Luchese, Rosa H; McIntosh, Douglas

    2015-06-01

    The occurrence of psychrotrophic bacteria in raw milk is studied worldwide due to the difficulties associated with controlling their growth during cold storage and the consequent negative effects upon fluid milk or dairy products. Among the psychrotrophic bacteria, the genus Pseudomonas (represented primarily by P. fluorescens) has been highlighted as the cause of numerous defects in dairy products. In light of its perceived predominance, this species has frequently been chosen as a model organism to assess the effects of psychrotrophic bacteria on milk or to evaluate the efficacy of control measures. However, recent findings derived from the application of molecular biological techniques have exposed a number of deficiencies in our knowledge of the biology of milk-associated psychrotrophs. Furthermore, it has been revealed that microbe to microbe communication plays a significant role in determining both the identities and the extent to which different groups of microbes develop during cold storage. The application of molecular identification methods has exposed errors in the classification of members of the genus Pseudomonas isolated from cold stored milk and has stimulated a reevaluation of the presumed status of P. fluorescens as the predominant milk-associated psychrotrophic species. This article presents a succinct review of data from studies on psychrotrophic bacteria in milk, some of which contest established theories in relation to the microbiology of cold stored raw milk, and poses the question: how much do we really know?

  6. DETECTION OF PHENOL DEGRADING BACTERIA AND PSEUDOMONAS PUTIDA IN ACTIVATED SLUDGE BY POLYMERASE CHAIN REACTION

    OpenAIRE

    H. Movahedyan ، H. Khorsandi ، R. Salehi ، M. Nikaeen

    2009-01-01

    Phenol is one of the organic pollutants in various industrial wastewaters especially petrochemical and oil refining. Biological treatment is one of the considerable choices for removing of phenol present in these wastewaters. Identification of effective microbial species is considered as one of the important priorities for production of the biomass in order to achieve desirable kinetic of biological reactions. Basic purpose of this research is identification of phenol-degrading Pseudomonas Pu...

  7. Evaluating the effect of some Pesodomonas bacteria strains on wheat yield and its components at various levels of phosphorus fertilization

    Directory of Open Access Journals (Sweden)

    Shahram Rezvan Beidokhti

    2016-04-01

    Full Text Available Application of biofertilizers, especially plant growth promoting rhizobacteria (PGPR is one of the most important strategies for plant nutrition compared to chemical fertilizers, especially in sustainable management of agroecosystems. In order to evaluate the effect of Pesodomonas bacteria strains and chemical phosphate fertilizer on yield and yield components of wheat the Kavir cultivar an experiment was conducted in experimental farm of the Agriculture Faculty of Azad University of Damghan. The treatments were arranged as split plots and were evaluated in a complete randomized block design with three replications. The chemical phosphate fertilizer levels of P0 (control, P1 (60 Kg/ha, P2 (90 Kg/ha, P3 (120 Kg/ha and P4 (150 Kg/ha of super phosphate triple were allocated to the main plots. While the different bacteria strains of Pseudomonas putida 21 (S1, Pseudomonas putida 158 (S2, Pseudomonas fluorescens 168 (S3 and non-inoculation control (S0 were allocated to the sub plots. The results of the experiment indicated that the highest grain yield of 7633 Kg/ha was obtained with application of 90 kg/ha of phosphorus fertilizer accompanied with S3 bacteria (strain No.168. The Pseudomonas fluorescens168 demonstrated a remarkable efficiency in dry matter and grain production in wheat with no chemical phosphate application.

  8. Interaction between fish spoilage bacteria Pseudomonas sp and Shewanella putrefaciens in fish extracts and on fish tissue

    DEFF Research Database (Denmark)

    Gram, Lone; Melchiorsen, Jette

    1996-01-01

    , supernatant fluids from siderophore- negative Pseudomonas isolates did not inhibit growth of S. putrefaciens. The inhibitory effect was, except for one strain of Pseudomonas, not seen in supernatant fluids from iron- enriched cultures of Pseudomonas sp. Finally, siderophore- producing Pseudomonas sp. lowered...

  9. Twenty-one genome sequences from Pseudomonas species and 19 genome sequences from diverse bacteria isolated from the rhizosphere and endosphere of Populus deltoides.

    Science.gov (United States)

    Brown, Steven D; Utturkar, Sagar M; Klingeman, Dawn M; Johnson, Courtney M; Martin, Stanton L; Land, Miriam L; Lu, Tse-Yuan S; Schadt, Christopher W; Doktycz, Mitchel J; Pelletier, Dale A

    2012-11-01

    To aid in the investigation of the Populus deltoides microbiome, we generated draft genome sequences for 21 Pseudomonas strains and 19 other diverse bacteria isolated from Populus deltoides roots. Genome sequences for isolates similar to Acidovorax, Bradyrhizobium, Brevibacillus, Caulobacter, Chryseobacterium, Flavobacterium, Herbaspirillum, Novosphingobium, Pantoea, Phyllobacterium, Polaromonas, Rhizobium, Sphingobium, and Variovorax were generated.

  10. Twenty-One Genome Sequences from Pseudomonas Species and 19 Genome Sequences from Diverse Bacteria Isolated from the Rhizosphere and Endosphere of Populus deltoides

    OpenAIRE

    Brown, Steven D.; Utturkar, Sagar M.; Klingeman, Dawn M.; Johnson, Courtney M.; Martin, Stanton L.; Land, Miriam L.; Lu, Tse-Yuan S.; Schadt, Christopher W.; Doktycz, Mitchel J.; Pelletier, Dale A.

    2012-01-01

    To aid in the investigation of the Populus deltoides microbiome, we generated draft genome sequences for 21 Pseudomonas strains and 19 other diverse bacteria isolated from Populus deltoides roots. Genome sequences for isolates similar to Acidovorax, Bradyrhizobium, Brevibacillus, Caulobacter, Chryseobacterium, Flavobacterium, Herbaspirillum, Novosphingobium, Pantoea, Phyllobacterium, Polaromonas, Rhizobium, Sphingobium, and Variovorax were generated.

  11. Twenty-One Genome Sequences from Pseudomonas Species and 19 Genome Sequences from Diverse Bacteria Isolated from the Rhizosphere and Endosphere of Populus deltoides

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Steven D [ORNL; Utturkar, Sagar M [ORNL; Klingeman, Dawn Marie [ORNL; Johnson, Courtney M [ORNL; Martin, Stanton [ORNL; Land, Miriam L [ORNL; Lu, Tse-Yuan [ORNL; Schadt, Christopher Warren [ORNL; Doktycz, Mitchel John [ORNL; Pelletier, Dale A [ORNL

    2012-01-01

    To aid in the investigation of the Populus deltoides microbiome we generated draft genome sequences for twenty one Pseudomonas and twenty one other diverse bacteria isolated from Populus deltoides roots. Genome sequences for isolates similar to Acidovorax, Bradyrhizobium, Brevibacillus, Burkholderia, Caulobacter, Chryseobacterium, Flavobacterium, Herbaspirillum, Novosphingobium, Pantoea, Phyllobacterium, Polaromonas, Rhizobium, Sphingobium and Variovorax were generated.

  12. Characterization of Bacteria Isolated from the Saffron (Crocus sativus L. Rhizosphere

    Directory of Open Access Journals (Sweden)

    Jami Al-Ahmadi Majid

    2017-12-01

    Full Text Available One purpose of assessing the soil alive and active community is the identification of beneficial bacteria to use them as biological fertilizers, replacing or supplementing synthetic fertilizers. Such biofertilizers are predicted for the sustainability of agricultural production, especially for low input systems such as saffron fields. The aim of this work was to isolate and identify saffron rhizobacteria and to evaluate their possible effects on saffron growth. During 2013/14, some bacteria were isolated from the rhizosphere of the saffron plantations of different age in Gol village, Birjand, Iran. In total, 12 bacteria species were identified based on phenotypic traits and 16S rDNA sequences analysis. The strains were identified as B. subtilis, B. anthracis, B. cereus, B. megaterium, Bacillus sp., Paenibacillus, Pseudomonas fluorescens, P. putida, Escherichia coli, Pectobacterium sp. and Pantoea sp., with the dominant population belonging to the genus Bacillus. In the field study, inoculation of soil with these strains did not affect the leaf dry weight of the cultivated saffron, however, the strains of P. fluorescens increased the leaf area while P. fluorescens, Paenibacillus, Pectobacterium and B. megaterium increased the number of daughter corms and Azotobacter, B. cereus, B. subtilis and B. megaterium increased the corm weight. Our finding revealed that some bacteria present in the soil of perennial saffron plantations have a promising potential for developing as a plant growth promoting rhizobacteria.

  13. Inhibition of quorum sensing in Pseudomonas aeruginosa biofilm bacteria by a halogenated furanone compound

    DEFF Research Database (Denmark)

    Hentzer, Morten; Riedel, K.; Rasmussen, Thomas Bovbjerg

    2002-01-01

    Novel molecular tools have been constructed which allow for in situ detection of N-acyl homoserine lactone (AHL)-mediated quorum sensing in Pseudomonas aeruginosa biofilms. The reporter responds to AHL activation of LasR lay expression of an unstable version of the green-fluorescent protein (Gfp......). Gfp-based reporter technology has been applied for non-destructive, single-cell-level detection of quorum sensing in laboratory-based P. aeruginosa biofilms. It is reported that a synthetic halogenated furanone compound, which is a derivative of the secondary metabolites produced by the Australian...... macroalga Delisea pulchra, is capable of interfering with AHL-mediated quorum sensing in P. aeruginosa. It is demonstrated that the furanone compound specifically represses expression of a PlasB-gfp reporter fusion without affecting growth or protein synthesis. In addition, it reduces the production...

  14. 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

  15. Biocontrol of tomato foot and root rot by Pseudomonas bacteria in stonewool

    NARCIS (Netherlands)

    Validov, Shamil

    2007-01-01

    The research described in this thesis shows that the enrichment technique based on competitive root tip colonization allows the isolation of bacteria which can protect plants from TFRR through the mechanism competition for nutrients (and niches). The efficacy of biocontrol of one of these strains,

  16. A Direct Quantitative Agar-Plate Based Assay for Analysis of Pseudomonas protegens PF-5 Degradation of Polyurethane Films (Postprint)

    Science.gov (United States)

    2014-10-02

    activity in Pseudomonas fluorescens CHA0. Mol. Plant - Microbe . Interact . 25, 1440e1449. Yang, X., Wang, S., Zhou, L., 2012. Effect of carbon source, C... Interactions between the Pf-5 colonies and thin polyurethane (PU) coatings on ZnSe coupons were evaluated for degradation using infrared spectroscopy...Small (1 mm diameter) colonies of Pseudomonas protegens Pf-5 (formerly Pseudomonas fluorescens Pf-5) were used for this work. Interactions between

  17. Growth Inhibition of Cocoa Pod Rot Fungus Phytophthora palmivora byPseudomonas fluorescence and Bacillus subtilis bacteria

    Directory of Open Access Journals (Sweden)

    Sakti Widyanta Pratama

    2013-08-01

    Full Text Available Black pod disease caused by Phytophthora palmivorafungus is one of the important diseases on cocoa crop. Pod rot is the most important disease because it may cause loss of cocoa pod. Until now, the fungal pathogen of cocoa black pod disease is still a crucial problem and there is no fungicide that is really effective against the disease. One alternative to control the cocoa black pod disease is by using biological agents as biofungicide, including utilizing Pseudomonas fluorescenceand Bacillus subtilis bacteria. The research was done by isolation of P. palmivora from infected pods of Kaliwining Experimental Station to obtain pure cultures of fungus and by multiplication of P. fluorescence and B. subtilis. Antagonist test was performed by inoculating P. palmivora into a petri dish in a distance of 3 cm from the edge. P. fluorescenceand B. Subtilis were inoculated into petridishes in three days after the fungal treatment. Control was inoculated with isolate of P. palmivora only. Fungal growth was measured everyday by measuring radius of fungal colonies first time 24 hours after inoculation. Growth of Phytophthora palmivora in the two treatmens were used to calculate the percentage of inhibition. The results of this study indicated that P. fluorescence and B. subtiliswere able to inhibit fungal growth of P. palmivora. Both bacterial antagonists had the same effectiveness in inhibiting the growth of P. palmivora fungus based on the percentage of inhibition and effectiveness criteria. Based on the results of translucent zones indicated that B. subtiliswas more powerfull in inhibiting growth of P. Palmivora compared to P. fluorescence. Key words: Black pod disease of cocoa, biological control, Phytophthora palmivora, Pseudomonas fluorescence, Bacillus subtilis

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

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    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.

  19. Combined Field Inoculations ofPseudomonasBacteria, Arbuscular Mycorrhizal Fungi, and Entomopathogenic Nematodes and their Effects on Wheat Performance.

    Science.gov (United States)

    Imperiali, Nicola; Chiriboga, Xavier; Schlaeppi, Klaus; Fesselet, Marie; Villacrés, Daniela; Jaffuel, Geoffrey; Bender, S Franz; Dennert, Francesca; Blanco-Pérez, Ruben; van der Heijden, Marcel G A; Maurhofer, Monika; Mascher, Fabio; Turlings, Ted C J; Keel, Christoph J; Campos-Herrera, Raquel

    2017-01-01

    In agricultural ecosystems, pest insects, pathogens, and reduced soil fertility pose major challenges to crop productivity and are responsible for significant yield losses worldwide. Management of belowground pests and diseases remains particularly challenging due to the complex nature of the soil and the limited reach of conventional agrochemicals. Boosting the presence of beneficial rhizosphere organisms is a potentially sustainable alternative and may help to optimize crop health and productivity. Field application of single beneficial soil organisms has shown satisfactory results under optimal conditions. This might be further enhanced by combining multiple beneficial soil organisms, but this remains poorly investigated. Here, we inoculated wheat plots with combinations of three beneficial soil organisms that have different rhizosphere functions and studied their effects on crop performance. Plant beneficial Pseudomonas bacteria, arbuscular mycorrhizal fungi (AMF), and entomopathogenic nematodes (EPN), were inoculated individually or in combinations at seeding, and their effects on plant performance were evaluated throughout the season. We used traditional and molecular identification tools to monitor their persistence over the cropping season in augmented and control treatments, and to estimate the possible displacement of native populations. In three separate trials, beneficial soil organisms were successfully introduced into the native populations and readily survived the field conditions. Various Pseudomonas , mycorrhiza, and nematode treatments improved plant health and productivity, while their combinations provided no significant additive or synergistic benefits compared to when applied alone. EPN application temporarily displaced some of the native EPN, but had no significant long-term effect on the associated food web. The strongest positive effect on wheat survival was observed for Pseudomonas and AMF during a season with heavy natural infestation by

  20. Combined Field Inoculations of Pseudomonas Bacteria, Arbuscular Mycorrhizal Fungi, and Entomopathogenic Nematodes and their Effects on Wheat Performance

    Directory of Open Access Journals (Sweden)

    Nicola Imperiali

    2017-10-01

    Full Text Available In agricultural ecosystems, pest insects, pathogens, and reduced soil fertility pose major challenges to crop productivity and are responsible for significant yield losses worldwide. Management of belowground pests and diseases remains particularly challenging due to the complex nature of the soil and the limited reach of conventional agrochemicals. Boosting the presence of beneficial rhizosphere organisms is a potentially sustainable alternative and may help to optimize crop health and productivity. Field application of single beneficial soil organisms has shown satisfactory results under optimal conditions. This might be further enhanced by combining multiple beneficial soil organisms, but this remains poorly investigated. Here, we inoculated wheat plots with combinations of three beneficial soil organisms that have different rhizosphere functions and studied their effects on crop performance. Plant beneficial Pseudomonas bacteria, arbuscular mycorrhizal fungi (AMF, and entomopathogenic nematodes (EPN, were inoculated individually or in combinations at seeding, and their effects on plant performance were evaluated throughout the season. We used traditional and molecular identification tools to monitor their persistence over the cropping season in augmented and control treatments, and to estimate the possible displacement of native populations. In three separate trials, beneficial soil organisms were successfully introduced into the native populations and readily survived the field conditions. Various Pseudomonas, mycorrhiza, and nematode treatments improved plant health and productivity, while their combinations provided no significant additive or synergistic benefits compared to when applied alone. EPN application temporarily displaced some of the native EPN, but had no significant long-term effect on the associated food web. The strongest positive effect on wheat survival was observed for Pseudomonas and AMF during a season with heavy

  1. Combined Field Inoculations of Pseudomonas Bacteria, Arbuscular Mycorrhizal Fungi, and Entomopathogenic Nematodes and their Effects on Wheat Performance

    Science.gov (United States)

    Imperiali, Nicola; Chiriboga, Xavier; Schlaeppi, Klaus; Fesselet, Marie; Villacrés, Daniela; Jaffuel, Geoffrey; Bender, S. Franz; Dennert, Francesca; Blanco-Pérez, Ruben; van der Heijden, Marcel G. A.; Maurhofer, Monika; Mascher, Fabio; Turlings, Ted C. J.; Keel, Christoph J.; Campos-Herrera, Raquel

    2017-01-01

    In agricultural ecosystems, pest insects, pathogens, and reduced soil fertility pose major challenges to crop productivity and are responsible for significant yield losses worldwide. Management of belowground pests and diseases remains particularly challenging due to the complex nature of the soil and the limited reach of conventional agrochemicals. Boosting the presence of beneficial rhizosphere organisms is a potentially sustainable alternative and may help to optimize crop health and productivity. Field application of single beneficial soil organisms has shown satisfactory results under optimal conditions. This might be further enhanced by combining multiple beneficial soil organisms, but this remains poorly investigated. Here, we inoculated wheat plots with combinations of three beneficial soil organisms that have different rhizosphere functions and studied their effects on crop performance. Plant beneficial Pseudomonas bacteria, arbuscular mycorrhizal fungi (AMF), and entomopathogenic nematodes (EPN), were inoculated individually or in combinations at seeding, and their effects on plant performance were evaluated throughout the season. We used traditional and molecular identification tools to monitor their persistence over the cropping season in augmented and control treatments, and to estimate the possible displacement of native populations. In three separate trials, beneficial soil organisms were successfully introduced into the native populations and readily survived the field conditions. Various Pseudomonas, mycorrhiza, and nematode treatments improved plant health and productivity, while their combinations provided no significant additive or synergistic benefits compared to when applied alone. EPN application temporarily displaced some of the native EPN, but had no significant long-term effect on the associated food web. The strongest positive effect on wheat survival was observed for Pseudomonas and AMF during a season with heavy natural infestation by

  2. Antifungal Activity of Selected Indigenous Pseudomonas and Bacillus from the Soybean Rhizosphere

    Science.gov (United States)

    León, M.; Yaryura, P. M.; Montecchia, M. S.; Hernández, A. I.; Correa, O. S.; Pucheu, N. L.; Kerber, N. L.; García, A. F.

    2009-01-01

    The purpose of this study was to isolate and select indigenous soil Pseudomonas and Bacillus bacteria capable of developing multiple mechanisms of action related to the biocontrol of phytopathogenic fungi affecting soybean crops. The screening procedure consisted of antagonism tests against a panel of phytopathogenic fungi, taxonomic identification, detection by PCR of several genes related to antifungal activity, in vitro detection of the antifungal products, and root colonization assays. Two isolates, identified and designated as Pseudomonas fluorescens BNM296 and Bacillus amyloliquefaciens BNM340, were selected for further studies. These isolates protected plants against the damping-off caused by Pythium ultimum and were able to increase the seedling emergence rate after inoculation of soybean seeds with each bacterium. Also, the shoot nitrogen content was higher in plants when seeds were inoculated with BNM296. The polyphasic approach of this work allowed us to select two indigenous bacterial strains that promoted the early development of soybean plants. PMID:20016811

  3. Antibiotic resistance patterns of Pseudomonas spp. isolated from the River Danube

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    Clemens eKittinger

    2016-05-01

    Full Text Available Spread and persistence of antibiotic resistance pose a severe threat to human health, yet there is still lack of knowledge about reservoirs of antibiotic resistant bacteria in the environment. We took the opportunity of the Joint Danube Survey 3 (JDS3, the world's biggest river research expedition of its kind in 2013, to analyse samples originating from different sampling points along the whole length of the river. Due to its high clinical relevance, we concentrated on the characterization of Pseudomonas spp. and evaluated the resistance profiles of Pseudomonas spp. which were isolated from eight sampling points. In total, 520 Pseudomonas isolates were found, 344 (66.0% isolates were identified as Pseudomonas putida, and 141 (27.1% as Pseudomonas fluorescens, all other Pseudomonas species were represented by less than five isolates, among those two P. aeruginosa isolates. Thirty seven percent (37% of all isolated Pseudomonas species showed resistance to at least one out of eleven tested antibiotics. The most common resistance was against meropenem (30.4% / 158 isolates piperacillin/tazobactam (10.6% / 55 isolates and ceftazidime (4.2% / 22 isolates. 16 isolates (3.1% / 16 isolates were multi-resistant. For each tested antibiotic at least one resistant isolate could be detected. Sampling points from the upper stretch of the River Danube showed more resistant isolates than downriver. Our results suggest that antibiotic resistance can be acquired by and persists even in Pseudomonas species that are normally not in direct contact with humans. A possible scenario is that these bacteria provide a reservoir of antibiotic resistance genes that can spread to related human pathogens by horizontal gene transfer.

  4. Isolation and characterization of heavy polycyclic aromatic hydrocarbon-degrading bacteria adapted to electrokinetic conditions.

    Science.gov (United States)

    Li, Fengmei; Guo, Shuhai; Hartog, Niels; Yuan, Ye; Yang, Xuelian

    2016-02-01

    Polycyclic aromatic hydrocarbon (PAH)-degrading bacteria capable of growing under electrokinetic conditions were isolated using an adjusted acclimation and enrichment procedure based on soil contaminated with heavy PAHs in the presence of an electric field. Their ability to degrade heavy PAHs under an electric field was individually investigated in artificially contaminated soils. The results showed that strains PB4 (Pseudomonas fluorescens) and FB6 (Kocuria sp.) were the most efficient heavy PAH degraders under electrokinetic conditions. They were re-inoculated into a polluted soil from an industrial site with a PAH concentration of 184.95 mg kg(-1). Compared to the experiments without an electric field, the degradation capability of Pseudomonas fluorescens and Kocuria sp. was enhanced in the industrially polluted soil under electrokinetic conditions. The degradation extents of total PAHs were increased by 15.4 and 14.0% in the electrokinetic PB4 and FB6 experiments (PB4 + EK and FB6 + EK) relative to the PB4 and FB6 experiments without electrokinetic conditions (PB4 and FB6), respectively. These results indicated that P. fluorescens and Kocuria sp. could efficiently degrade heavy PAHs under electrokinetic conditions and have the potential to be used for the electro-bioremediation of PAH-contaminated soil, especially if the soil is contaminated with heavy PAHs.

  5. Mannitol enhances antibiotic sensitivity of persister bacteria in Pseudomonas aeruginosa biofilms.

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    Nicolas Barraud

    Full Text Available The failure of antibiotic therapies to clear Pseudomonas aeruginosa lung infection, the key mortality factor for cystic fibrosis (CF patients, is partly attributed to the high tolerance of P. aeruginosa biofilms. Mannitol has previously been found to restore aminoglycoside sensitivity in Escherichia coli by generating a proton-motive force (PMF, suggesting a potential new strategy to improve antibiotic therapy and reduce disease progression in CF. Here, we used the commonly prescribed aminoglycoside tobramycin to select for P. aeruginosa persister cells during biofilm growth. Incubation with mannitol (10-40 mM increased tobramycin sensitivity of persister cells up to 1,000-fold. Addition of mannitol to pre-grown biofilms was able to revert the persister phenotype and improve the efficacy of tobramycin. This effect was blocked by the addition of a PMF inhibitor or in a P. aeruginosa mutant strain unable to metabolise mannitol. Addition of glucose and NaCl at high osmolarity also improved the efficacy of tobramycin although to a lesser extent compared to mannitol. Therefore, the primary effect of mannitol in reverting biofilm associated persister cells appears to be an active, physiological response, associated with a minor contribution of osmotic stress. Mannitol was tested against clinically relevant strains, showing that biofilms containing a subpopulation of persister cells are better killed in the presence of mannitol, but a clinical strain with a high resistance to tobramycin was not affected by mannitol. Overall, these results suggest that in addition to improvements in lung function by facilitating mucus clearance in CF, mannitol also affects antibiotic sensitivity in biofilms and does so through an active, physiological response.

  6. Mannitol Enhances Antibiotic Sensitivity of Persister Bacteria in Pseudomonas aeruginosa Biofilms

    Science.gov (United States)

    Barraud, Nicolas; Buson, Alberto; Jarolimek, Wolfgang; Rice, Scott A.

    2013-01-01

    The failure of antibiotic therapies to clear Pseudomonas aeruginosa lung infection, the key mortality factor for cystic fibrosis (CF) patients, is partly attributed to the high tolerance of P. aeruginosa biofilms. Mannitol has previously been found to restore aminoglycoside sensitivity in Escherichia coli by generating a proton-motive force (PMF), suggesting a potential new strategy to improve antibiotic therapy and reduce disease progression in CF. Here, we used the commonly prescribed aminoglycoside tobramycin to select for P. aeruginosa persister cells during biofilm growth. Incubation with mannitol (10–40 mM) increased tobramycin sensitivity of persister cells up to 1,000-fold. Addition of mannitol to pre-grown biofilms was able to revert the persister phenotype and improve the efficacy of tobramycin. This effect was blocked by the addition of a PMF inhibitor or in a P. aeruginosa mutant strain unable to metabolise mannitol. Addition of glucose and NaCl at high osmolarity also improved the efficacy of tobramycin although to a lesser extent compared to mannitol. Therefore, the primary effect of mannitol in reverting biofilm associated persister cells appears to be an active, physiological response, associated with a minor contribution of osmotic stress. Mannitol was tested against clinically relevant strains, showing that biofilms containing a subpopulation of persister cells are better killed in the presence of mannitol, but a clinical strain with a high resistance to tobramycin was not affected by mannitol. Overall, these results suggest that in addition to improvements in lung function by facilitating mucus clearance in CF, mannitol also affects antibiotic sensitivity in biofilms and does so through an active, physiological response. PMID:24349568

  7. Water-soluble Moringa oleifera lectin interferes with growth, survival and cell permeability of corrosive and pathogenic bacteria.

    Science.gov (United States)

    Moura, M C; Napoleão, T H; Coriolano, M C; Paiva, P M G; Figueiredo, R C B Q; Coelho, L C B B

    2015-09-01

    This work evaluated the antibacterial activity of a water-soluble Moringa oleifera seed lectin (WSMoL) by evaluating its effect on growth, survival and cell permeability of Bacillus sp., Bacillus cereus, Bacillus pumillus, Bacillus megaterium, Micrococcus sp., Pseudomonas sp., Pseudomonas fluorescens, Pseudomonas stutzeri and Serratia marcescens. In addition, the effect of lectin on membrane integrity of most sensitive species was also evaluated. All the tested bacteria are able to cause biocorrosion and some are also responsible for human infections. WSMoL inhibited the bacterial growth, induced agglutination and promoted the leakage of proteins from cells of all strains. Bactericidal effect was detected against Bacillus sp., B. pumillus, B. megaterium, Ps. fluorescens and Ser. marcescens. The bacteriostatic effect of lectin was evident with only 6 h of incubation. Fluorescence microscopy of Ser. marcescens showed that WSMoL caused loss of cell integrity and indicated an anti-biofilm activity of the lectin. WSMoL was active against the bacteria by inhibiting growth and affecting cell permeability. The lectin also interfered with membrane integrity of Ser. marcescens, the most sensitive species. The study indicates that WSMoL was active against bacteria that cause serious problems in both industrial and health sectors. Also, the study contributes for the 'state-of-art' on antibacterial mechanisms of lectins. © 2015 The Society for Applied Microbiology.

  8. The Application of PCR Reaction for Identification of MHB Bacteria Species

    Directory of Open Access Journals (Sweden)

    Ząbkiewicz Anna

    2014-07-01

    Full Text Available This study characterizes mycorrhiza helper bacteria (MHB from selected unpolluted locations as well as subjected to industrial emissions. To determine the species of bacteria isolated from the roots of ectomycorrhizal pine and birch, a method based on the sequence analysis of a 16S rRNA gene was used. The isolated bacteria were initially characterized by available biochemical methods and phenotypic observation. On the selected bacteria representatives isolation of DNA was performed, on which the PCR reaction was carried out. In this way amplified samples were automatically sequenced and the obtained results were compared to public databases. Among the isolated bacteria Pseudomonas fluorescens SBW25 and Burkholderia xenovorans LB400 species were dominant.

  9. Phenotypic Characterization of Pseudomonas Bacteria Isolated from Polluted Sites of Ostrava, Czech Republic /

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    Vojtková Hana

    2012-09-01

    Full Text Available Životní prostředí v průmyslových městech je obvykle vystaveno vysokým vlivům antropogenní kontaminace pocházející zejména ze spalování fosilních paliv, silniční dopravy, průmyslové činnosti, těžby a zpracování rud. Degradační potenciál kontaminovaných půd je výrazně ovlivněn diverzitou a funkcí mikrobiálního ekosystému dané lokality, v jednotlivých společenstvech lze diagnostikovat shodné i rozdílné vlastnosti specifických bakteriálních kmenů. Z reálných půdních vzorků z území města Ostravy, z lokalit zatížených průmyslovou činností, byly izolovány nové bakteriální kmeny, které byly srovnávány na základě výsledků biochemické identifikace pomocí moderního systému BIOLOG MicroStation. U nově izolovaných kmenů jsou uvedeny jejich základní charakteristiky, které dokládají fenotypovou variabilitu nejen na úrovni rodu Pseudomonas, ale také na úrovni jednotlivých druhů. Rozdíly ve fyziologických parametrech izolovaných kmenů jsou dány do souvislosti se schopností adaptace půdních mikroorganismů na podmínky znečištěného prostředí. Článek podává přehled o významu fenotypové charakteristiky pro identifikaci mikroorganismů a jejich správného taxonomického řazení.

  10. Effects of inoculation with organic-phosphorus-mineralizing bacteria on soybean (Glycine max) growth and indigenous bacterial community diversity.

    Science.gov (United States)

    Sun, Wei; Qian, Xun; Gu, Jie; Wang, Xiao-Juan; Li, Yang; Duan, Man-Li

    2017-05-01

    Three different organic-phosphorus-mineralizing bacteria (OPMB) strains were inoculated to soil planted with soybean (Glycine max), and their effects on soybean growth and indigenous bacterial community diversity were investigated. Inoculation with Pseudomonas fluorescens Z4-1 and Brevibacillus agri L7-1 increased organic phosphorus degradation by 22% and 30%, respectively, compared with the control at the mature stage. Strains P. fluorescens Z4-1 and B. agri L7-1 significantly improved the soil alkaline phosphatase activity, average well color development, and the soybean root activity. Terminal restriction fragment length polymorphism analysis demonstrated that P. fluorescens Z4-1 and B. agri L7-1 could persist in the soil at relative abundances of 2.0%-6.4% throughout soybean growth. Thus, P. fluorescens Z4-1 and B. agri L7-1 could potentially be used in organic-phosphorus-mineralizing biofertilizers. OPMB inoculation altered the genetic structure of the soil bacterial communities but had no apparent influence on the carbon source utilization profiles of the soil bacterial communities. Principal components analysis showed that the changes in the carbon source utilization profiles of bacterial community depended mainly on the plant growth stages rather than inoculation with OPMB. The results help to understand the evolution of the soil bacterial community after OPMB inoculation.

  11. 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

  12. Bacteria in the leaf ecosystem with emphasis on Pseudomonas syringae-a pathogen, ice nucleus, and epiphyte.

    Science.gov (United States)

    Hirano, S S; Upper, C D

    2000-09-01

    The extremely large number of leaves produced by terrestrial and aquatic plants provide habitats for colonization by a diversity of microorganisms. This review focuses on the bacterial component of leaf microbial communities, with emphasis on Pseudomonas syringae-a species that participates in leaf ecosystems as a pathogen, ice nucleus, and epiphyte. Among the diversity of bacteria that colonize leaves, none has received wider attention than P. syringae, as it gained notoriety for being the first recombinant organism (Ice(-) P. syringae) to be deliberately introduced into the environment. We focus on P. syringae to illustrate the attractiveness and somewhat unique opportunities provided by leaf ecosystems for addressing fundamental questions of microbial population dynamics and mechanisms of plant-bacterium interactions. Leaf ecosystems are dynamic and ephemeral. The physical environment surrounding phyllosphere microbes changes continuously with daily cycles in temperature, radiation, relative humidity, wind velocity, and leaf wetness. Slightly longer-term changes occur as weather systems pass. Seasonal climatic changes impose still a longer cycle. The physical and physiological characteristics of leaves change as they expand, mature, and senesce and as host phenology changes. Many of these factors influence the development of populations of P. syringae upon populations of leaves. P. syringae was first studied for its ability to cause disease on plants. However, disease causation is but one aspect of its life strategy. The bacterium can be found in association with healthy leaves, growing and surviving for many generations on the surfaces of leaves as an epiphyte. A number of genes and traits have been identified that contribute to the fitness of P. syringae in the phyllosphere. While still in their infancy, such research efforts demonstrate that the P. syringae-leaf ecosystem is a particularly attractive system with which to bridge the gap between what is known

  13. 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.)

  14. Isolation, characterization and phylogeny of sponge-associated bacteria with antimicrobial activities from Brazil.

    Science.gov (United States)

    Santos, Olinda C S; Pontes, Paula V M L; Santos, Juliana F M; Muricy, Guilherme; Giambiagi-deMarval, Marcia; Laport, Marinella S

    2010-09-01

    Bacteria associated with marine sponges represent a rich source of bioactive metabolites. The aim of this study was to isolate and characterize bacteria with antimicrobial activities from Brazilian sponges. A total of 158 colony-forming units were isolated from nine sponge species. Among these, 12 isolates presented antimicrobial activities against pathogenic bacteria. Based on comparative sequence analysis of their 16S rRNA genes, the sponge-associated bacterial strains could be subdivided into three phylogenetically different clusters. Five strains were affiliated with Firmicutes (genera Bacillus and Virgibacillus), three with alpha-Proteobacteria (Pseudovibrio sp.) and four with gamma-Proteobacteria (genera Pseudomonas and Stenotrophomonas). The sponge-associated bacterial strains Pseudomonas fluorescens H40 and H41 and Pseudomonas aeruginosa H51 exhibited antimicrobial activity against both Gram-negative and Gram-positive bacteria, including strains such as vancomycin-resistant Enterococcus faecium and multiresistant Klebsiella pneumoniae. Bacillus pumilus Pc31 and Pc32, Pseudovibrio ascidiaceicola Pm31 and Ca31 and Pseudovibrio denitrificans Mm37 strains were more effective against Gram-positive bacteria. These findings suggest that the identified strains may contribute to the search for new sources of antimicrobial substances, an important strategy for developing alternative therapies to treat infections caused by multidrug-resistant bacteria. Copyright 2010 Elsevier Masson SAS. All rights reserved.

  15. An epifluorescence-based evaluation of the effects of short-term particle association on the chlorination of surface water bacteria.

    Science.gov (United States)

    Lynch, Fiona; Tomlinson, Steven; Palombo, Enzo A; Harding, Ian H

    2014-10-15

    Investigations into particle-mediated chlorination resistance were undertaken for three different bacteria (Escherichia coli ATCC 25922 and environmental isolates of Pseudomonas fluorescens and Serratia marcescens) and three different surfaces (goethite, environmental particles and surface-modified environmental particles). P. fluorescens demonstrated greater hydrophobicity than both other strains and proved the most adherent bacterium over all substrata investigated. Particle-mediated resistance to chlorination was investigated using short bacteria-particle association times and activity assays that employed sensitive epifluorescent detection. Consistent with adhesive behaviours, the bacterial strain that demonstrated the greatest particle-mediated chlorination resistance was the environmental strain of P. fluorescens. Resistance was observed to vary with both bacteria and particle type, and demonstrated a moderate correlation with adhesion (r(2) ≥ 0.65). The short-term approach employed in our study demonstrates particle-mediated protection without the commonly assumed requirements of extracellular polymeric substances (EPS) or a large particle-based chlorine demand. Consequently, we have linked resistance with adhesion capacities and demonstrated a limit to resistance in the presence of additional particle protective sites (through increased turbidity) which appears to be driven by intra-population variance in bacterial surface characteristics. Finally, we observed important differences between behaviours of environmental versus laboratory-derived bacterial strains and particles, which highlight the importance of employing both approaches in characterising "real world" systems. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Influence of volatile organic compounds emitted by Pseudomonas and Serratia strains on Agrobacterium tumefaciens biofilms.

    Science.gov (United States)

    Plyuta, Vladimir; Lipasova, Valentina; Popova, Alexandra; Koksharova, Olga; Kuznetsov, Alexander; Szegedi, Erno; Chernin, Leonid; Khmel, Inessa

    2016-07-01

    The ability to form biofilms plays an important role in bacteria-host interactions, including plant pathogenicity. In this work, we investigated the action of volatile organic compounds (VOCs) produced by rhizospheric strains of Pseudomonas chlororaphis 449, Pseudomonas fluorescens B-4117, Serratia plymuthica IC1270, as well as Serratia proteamaculans strain 94, isolated from spoiled meat, on biofilms formation by three strains of Agrobacterium tumefaciens which are causative agents of crown-gall disease in a wide range of plants. In dual culture assays, the pool of volatiles emitted by the tested Pseudomonas and Serratia strains suppressed the formation of biofilms of A. tumefaciens strains grown on polycarbonate membrane filters and killed Agrobacterium cells in mature biofilms. The individual VOCs produced by the tested Pseudomonas strains, that is, ketones (2-nonanone, 2-heptanone, 2-undecanone), and dimethyl disulfide (DMDS) produced by Serratia strains, were shown to kill A. tumefaciens cells in mature biofilms and suppress their formation. The data obtained in this study suggest an additional potential of some ketones and DMDS as protectors of plants against A. tumefaciens strains, whose virulence is associated with the formation of biofilms on the infected plants. © 2016 APMIS. Published by John Wiley & Sons Ltd.

  17. Multistep introduction of bacteria to natural substrates at different initial inoculation doses

    Science.gov (United States)

    Kupriyanov, A. A.; Semenov, A. M.; Kunenkova, N. N.; van Bruggen, A. H. C.

    2009-09-01

    The population dynamics of the saprotrophic Pseudomonas fluorescens 32 gfp bacteria and two conventionally pathogenic enterobacteria ( Escherichia coli 0157:H7 and Salmonella enterica var. Typhimurium) were investigated in their inoculation at different doses into cattle excreta and their subsequent entering soil and plants and migration through the gastroenteric tract of invertebrates. All the introduced bacteria investigated are shown to be able to overcome ecological barriers as they migrate through the natural substrates and habitats. The introduce microorganisms maintain their high population density even at the lowest initial inoculation dose—105 CFU/g of dry matter. Plants were found to be a favorable substrate for the survival of the bacteria investigated (for enteropathogens, in particular). Enteropathogens are able to pass through the gastroenteric tract of invertebrates. Therefore, these organisms can function as incubators and carriers of enteroinfections in nature.

  18. Fungi and bacteria associated with the wet and brown wood in trunk of Betula pendula trees

    Directory of Open Access Journals (Sweden)

    Krystyna Przybył

    2014-01-01

    Full Text Available Bacteria and fungi were isolated from external and internal zone of brown and water saturated wood of trunk of Betula pendula trees aged 44-46. Quantitative and qualitative differences in the bacterial and fungal populations were found between both the zones. Populations of bacteria increased towards the internal sapwood, contrary to the fungi which more frequently colonised the external zone. The most common bacteria were Pseudomonas spp. (mainly P.fluorescens biovar I whereas Bacillus macerans, B. alvei and Erwinia heibicola were able to degrade the polygalacturonic acid and pectin gels. In case of the fungi population, the most common (more than 3% colonising the external zone were successively: Phialophora fastigiata, Trichoderma harzianum, Alternaria alternata, Mortierella isabellina, Cladosporium herbarum, T. viride, C. cladosporioides and Melanconium betulinum. In the community of fungi occurring in the internal zone, the most common (more than 6.5% were:Cladosporium herbarum, Phialophora lagerbergii and Ph.fastigiata.

  19. Virulence of entomopathogenic bacteria in the bed bug, Cimex lectularius.

    Science.gov (United States)

    Pietri, Jose E; Liang, Dangsheng

    2017-10-24

    Due in part to the development of insecticide resistance, the common bed bug, Cimex lectularius, has overcome human intervention efforts to make a global resurgence. The failure of chemical pesticides has created a need for novel strategies to combat bed bugs. While a number of insect pests are susceptible to the use of entomopathogenic microbes or microbial-derived toxins, biological control methods have not been thoroughly explored in bed bugs. Here, we tested the virulence of three entomopathogenic bacterial species in C. lectularius to determine their potential for bed bug control. We examined bed bug survival after inoculation with live or heat-killed Serratia marcescens, Pseudomonas fluorescens, and Bacillus thuringiensis israelensis at varying temperatures. We also analyzed the viability and growth of the same bacteria in infected bed bugs. All three bacterial species were pathogenic to bed bugs. However, the effects of S. marcescens and P. fluorescens were temperature-dependent while the lethality of B. thuringiensis israelensis was not. In addition, bacterial virulence was partly dependent on the route of infection but was not strongly associated with proliferation. Thus, our results suggest multiple possible mechanisms of microbial pathogenicity in the bed bug and indicate that entomopathogenic bacteria, or products derived from them, may have useful applications for bed bug control. Copyright © 2017 Elsevier Inc. All rights reserved.

  20. Survival of Salmonella Typhimurium on soybean sprouts after treatment with gaseous chlorine dioxide and biocontrol Pseudomonas bacteria

    Science.gov (United States)

    Control of Salmonella Typhimurium on sprouts and minimally processed produce is crucial for food and consumer safety. The aim of this research was to assess natural microflora populations on soybean and evaluate the effects of gaseous chlorine dioxide (ClO2) and biocontrol Pseudomonas on the surviva...

  1. Survival of Salmonella enterica on soybean sprouts following treatments with gaseous chlorine dioxide and biocontrol Pseudomonas bacteria

    Science.gov (United States)

    Control of Salmonella enterica on sprouts and minimally processed, ready-to-eat fruits and vegetables is important for food and consumer safety. The aim of this research was to assess the effects of gaseous chlorine dioxide(ClO2)and biocontrol microorganisms (Pseudomonas chlororaphis and P. fluoresc...

  2. Detecção de bacteriocinas produzidas por bactérias fitopatogênicas dos gêneros Erwinia, Pseudomonas e Xanthomonas Detection of bacteriocins produced by plant pathogenic bacteria from the genera Erwinia, Pseudomonas and Xanthomonas

    Directory of Open Access Journals (Sweden)

    C.M.R. de Biagi

    1992-01-01

    Full Text Available A detecção da produção de bacteriocinas foi estudada em diferentes condições, utilizando-se linhagens de bactérias fitopatogênicas dos gêneros Erwinia, Pseudomonas e Xanthomonas. Obtiveram-se, respectivamente, 58,06%, 71,31% e 40,00% de linhagens produtoras no meio 523, que foi o mais indicado para detecção dessa produção. Uma maior concentração de ágar adicionada ao meio (1,5% produziu melhores resultados mas, a quantidade de meio nas placas não influenciou a detecção de bacteriocinas. O maior tempo de incubação utilizado (72 h. facilitou detecção de halos de produção de bacteriocinas. Irradiação com luz ultra-violeta em baixas doses parece facilitar a liberação de bacteriocinas, mas isso varia com a linhagem empregada.Detection of bacteriocin production was studied under distinct conditions using strains of plant pathogenic bacteria from the genera Erwinia, Pseudomonas and Xanthomonas. 58.06%, 79.31% and 40.00% of producing strains were found respectivelly in the three groups of bacteria using the 523 medium which was the best for the detection of bacteriocin production. Increasing agar concentrations added to the medium up to 1,5% improved the detection. The amount of medium added to the Petri dishes did not affect bacteriocin production. The longest incubation time (72h. improved the detection of haloe production. Ultra-violet irradiation in low dosages seems to improve the visualization of haloe prodution but this is dependent on the tested strains.

  3. Antimicrobial activities of commercial essential oils and their components against food-borne pathogens and food spoilage bacteria

    Science.gov (United States)

    Mith, Hasika; Duré, Rémi; Delcenserie, Véronique; Zhiri, Abdesselam; Daube, Georges; Clinquart, Antoine

    2014-01-01

    This study was undertaken to determine the in vitro antimicrobial activities of 15 commercial essential oils and their main components in order to pre-select candidates for potential application in highly perishable food preservation. The antibacterial effects against food-borne pathogenic bacteria (Listeria monocytogenes, Salmonella Typhimurium, and enterohemorrhagic Escherichia coli O157:H7) and food spoilage bacteria (Brochothrix thermosphacta and Pseudomonas fluorescens) were tested using paper disk diffusion method, followed by determination of minimum inhibitory (MIC) and bactericidal (MBC) concentrations. Most of the tested essential oils exhibited antimicrobial activity against all tested bacteria, except galangal oil. The essential oils of cinnamon, oregano, and thyme showed strong antimicrobial activities with MIC ≥ 0.125 μL/mL and MBC ≥ 0.25 μL/mL. Among tested bacteria, P. fluorescens was the most resistant to selected essential oils with MICs and MBCs of 1 μL/mL. The results suggest that the activity of the essential oils of cinnamon, oregano, thyme, and clove can be attributed to the existence mostly of cinnamaldehyde, carvacrol, thymol, and eugenol, which appear to possess similar activities against all the tested bacteria. These materials could be served as an important natural alternative to prevent bacterial growth in food products. PMID:25473498

  4. Novel Pseudomonas fluorescens Septic Sacroiliitis in a Healthy Soldier

    Science.gov (United States)

    2013-08-01

    pain and intermittent non-bloody diarrhea . Laboratory studies were unremarkable, concurrent abdominal computed tomography (CT) and lumbar MRI showed no...Microbiol 2008; 8: 189–202. 4. Dubey L, Krasinski K, Hernanz-Schulman M: Osteomyelitis secondary to trauma or infected contiguous soft tissue. Pediatr ...51. 17. Wu M-S, Chang S-S, Lee S-H, Lee C-C: Pyogenic sacroiliitis—a com- parison between pediatric and adult patients. Rheumatology 2007; 46: 1684–7

  5. Bacterial effects on arbuscular mycorrhizal fungi and mycorrhiza development as influenced by the bacteria, fungi, and host plant.

    Science.gov (United States)

    Pivato, Barbara; Offre, Pierre; Marchelli, Sara; Barbonaglia, Bruno; Mougel, Christophe; Lemanceau, Philippe; Berta, Graziella

    2009-02-01

    Bacterial strains from mycorrhizal roots (three belonging to Comamonadaceae and one to Oxalobacteraceae) and from non-mycorrhizal roots (two belonging to Comamonadaceae) of Medicago truncatula and two reference strains (Collimonas fungivorans Ter331 and Pseudomonas fluorescens C7R12) were tested for their effect on the in vitro saprophytic growth of Glomus mosseae BEG12 and on its colonization of M. truncatula roots. Only the Oxalobacteraceae strain, isolated from barrel medic mycorrhizal roots, and the reference strain P. fluorescens C7R12 promoted both the saprophytic growth and root colonization of G. mosseae BEG12, indicating that they acted as mycorrhiza helper bacteria. Greatest effects were achieved by P. fluorescens C7R12 and its influence on the saprophytic growth of G. mosseae was compared to that on Gigaspora rosea BEG9 to determine if the bacterial stimulation was fungal specific. This fungal specificity, together with plant specificity, was finally evaluated by comparing bacterial effects on arbuscular mycorrhizal symbiosis when each of the fungal species was inoculated to two different plant species (M. truncatula and Lycopersicon esculentum). The results obtained showed that promotion of saprophytic growth by P. fluorescens C7R12 was expressed in vitro towards G. mosseae but not towards G. rosea. Bacterial promotion of mycorhization was also expressed towards G. mosseae, but not G. rosea, in roots of M. truncatula and L. esculentum. Taken together, results indicated that enhancement of arbuscular mycorrhiza development was only induced by a limited number of bacteria, promotion by the most efficient bacterial strain being fungal and not plant specific.

  6. 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

  7. Impact of the microscale distribution of a Pseudomonas strain introduced into soil on potential contacts with indigenous bacteria

    DEFF Research Database (Denmark)

    Dechesne, Arnaud; Pallud, C.; Bertolla, F.

    2005-01-01

    Soil bioaugmentation is a promising approach in soil bioremediation and agriculture. Nevertheless, our knowledge of the fate and activity of introduced bacteria in soil and thus of their impact on the soil environment is still limited. The microscale spatial distribution of introduced bacteria has...

  8. Olivine dissolution in the presence of heterotrophic bacteria (Pseudomonas reactants) extracted from Icelandic groundwater of the CO2 injection pilot site

    Science.gov (United States)

    Shirokova, Liudmila; Pokrovsky, Oleg; Benezeth, Pascale; Gerard, Emmanuelle; Menez, Benedicte; Alfredsson, Helgi

    2010-05-01

    This work is aimed at experimental modeling of the effect of heterotrophic bacteria on dissolution of important rock-forming mineral, olivine, at the conditions of CO2 storage and sequestration. Heterotrophic aerobic gram-negative bacteria were extracted from deep underground water (HK31, 1700 m deep and, t = 25-30°C) of basaltic aquifer located within the Hellisheidi CO2 injection pilot site (Iceland). Following this sampling, we separated, using culture on nutrient agar plates, four different groups of gram-negative aerobic bacteria. The enzymatic activity of studied species has been evaluated using Biolog Ecoplates and their genetic identification was performed using 18-S RNA analysis. The optimal growth conditions of bacteria on Brain Hearth Broth nutrient have been determined as 5 to 37°C and growth media pH varied from 7.0-8.2. Culturing experiments allowed determining the optimal physico-chemical conditions for bacteria experiments in the presence of basic Ca, Mg-containing silicates. Olivine (Fo92) was chosen as typical mineral of basalt, widely considered in carbon dioxide sequestration mechanisms. Dissolution experiments were performed in constant-pH (7 to 9), bicarbonate-buffered (0.001 to 0.05 M) nutrient-diluted media in batch reactors at 0-30 bars of CO2 in the presence of various biomass of Pseudomonas reactants. The release rate of magnesium, silica and iron was measured as a function of time in the presence of live, actively growing, dead (autoclaved or glutaraldehyde-treated) cells and bacteria exometabolites. Both nutrient media diluted 10 times (to 100 mg DOC/L) and inert electrolyte (NaCl, no DOC) were used. Our preliminary results indicate that the pH and dissolved organic matter are the first-order parameters that control the element release from olivine at far from equilibrium conditions. The SEM investigation of reacted surfaces reveal formation of surface roughness with much stronger mineral alteration in the presence of live bacteria

  9. 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.

  10. Microbial Adhesion to Processing Lines for Fish Fillets and Cooked Shrimp: Influence of Stainless Steel Surface Finish and Presence of Gram-Negative Bacteria on the Attachment of Listeria monocytogenes

    Directory of Open Access Journals (Sweden)

    Hjörleifur Einarsson

    2005-01-01

    Full Text Available Microflora adhering to surfaces of processing lines in a shrimp factory and a fish processing plant was identified in situ and adhesion of mixed culture of Listeria monocytogenes and Gram-negative bacteria on stainless steel surfaces (untreated, polished and glass beaded was studied ex situ. The predominant genus attached to the surfaces was Pseudomonas spp. (66 % in the shrimp factory and Enterobacteriaceae (27 % in the fish factory. Shrimp juice was used as an enrichment broth during the study of adhered bacteria. Three different Gram-negative strains and a mixture of Pseudomonas spp. were selected to study their attachment together with L. monocytogenes to stainless steel surfaces. Highest numbers of the attached bacteria were obtained after the contamination with a mixed culture of L. monocytogenes and Serratia liquefaciens. A lower number of bacteria adhered to stainless steel surfaces when mixed cultures of L. monocytogenes and Pseudomonas fluorescens or Aeromonas spp. were tested. No significant differences (p<0.05 were observed in the bacteria attached to differently treated steel surfaces with different roughness (Ra=0.1–0.8 m. Bacterial adhesion increased with longer contact time. Colonisation of L. monocytogenes on stainless steel surfaces was significantly enhanced only in the presence of mixed Pseudomonas spp. These results indicate that smooth surfaces do not necessarily provide hygiene benefits over rougher surfaces.

  11. Translocation of bacteria from animal excrements to soil and associated habitats

    Science.gov (United States)

    Kupriyanov, A. A.; Kunenkova, N. N.; van Bruggen, A. H. C.; Semenov, A. M.

    2009-11-01

    The population dynamics of Salmonella enterica var. Typhimurium MAE 110 gfp, Escherichia coli O157:H7 gfp, and Pseudomonas fluorescens 32 gfp were investigated in their introduction to cattle excrements and subsequent entering the soil, plants of cress ( Lepidium sativum L.), and migration through the gastroenteric tract of French snails ( Helix pomatia L.). The survival of these bacteria in the excrements and soil was investigated at cyclically changing (day-night, 25-15 °C) and constant (18 °C) temperatures. The cyclically changing temperature adversely affected the survival of E. coli O157:H7 gfp, and P. fluorescens but did not influence S. enterica var. Typhimurium. All the bacteria and, especially, the analogues of enteropathogens showed high survival in the cattle and snail excrements, soil, and on the plants under the gradual decrease in their population. On the cress plants grown in a mixture of cattle excrements and soil, an increase in the number of the introduced bacteria was observed.

  12. Commensal Pseudomonas Species Isolated from Wastewater and Freshwater Milieus in the Eastern Cape Province, South Africa, as Reservoir of Antibiotic Resistant Determinants

    Directory of Open Access Journals (Sweden)

    Anthony I. Okoh

    2012-07-01

    Full Text Available Pseudomonas species are opportunistic pathogens with implications in a wide range of diseases including cystic fibrosis and sickle cell anaemia. Because of their status as multidrug resistant (MDR and extremely drug resistant (XDR bacteria Pseudomonas species represent a threat to public health. Prevalence, antibiogram and associated antibiotic resistant genes of Pseudomonas species isolated from freshwater and mixed liquor environments in the Eastern Cape Province of South Africa were assessed. Polymerase chain reaction (PCR based technique was used to identify the isolates and screen for antibiotic resistant genes. The result shows occurrence of Pseudomonas spp. in freshwater and mixed liquor as follows: 71.42% and 37.5% (P. putida, 14.28% and 31.25% (P. flourescens, 7.14% and 6.25% (P. aeruginosa and 7.14% and 25% for other Pseudomonas species respectively. Disk diffusion antibiogram of the Pseudomonas isolates from the two locations showed 100% resistance to penicillin, oxacillin, clindamycin, rifampicin and 100% susceptibility to ciprofloxacin and gentamicin with varied percentage resistances to cephalothin, nalidixic acid, tetracycline, and ampicillin. The blaTEM antibiotic resistant gene was detected in 12.5% of P. putida, 57.14% of P. fluorescens, 100% P. aeruginosa and 40% in other Pseudomonas species. Similarly, Integrons conserved segment were detected in 12.5% of P. putida, 57.14% of P. fluorescens, 100% of P. aeruginosa and 40% of other Pseudomonas species. The presence of blaTEM gene and integrons conserved segment in some of the isolates is worrisome and suggest Pseudomonas species as important reservoirs of multidrug resistance genes in the Eastern Cape Province environment.

  13. Colony to colorimetry in 6 h: ELISA detection of a surface-expressed Pseudomonas aeruginosa virulence factor using immobilized bacteria.

    Science.gov (United States)

    Adawi, Azmi; Neville, Lewis F

    2012-09-01

    A rapid ELISA employing intact Pseudomonas aeruginosa (PA) is described that allows discrimination between strains harboring flagellin type a or b. All 52 PA strains known to harbor flagellin type b were positive in this ELISA when screened with a fully human monoclonal antibody (LST-007) targeting flagellin type b. Completion of this assay in only 6 h, from picking a single bacterial colony to a colorimetric product, could easily be adapted to a clinical laboratory setting and permit the appropriate choice of therapeutic monoclonal antibody versus its homologous flagellin target in PA-infected patients. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. Antimicrobial multiresistance in bacteria isolated from freshwater Chilean salmon farms.

    Science.gov (United States)

    Mirand, Claudio D; Zemelman, Raul

    2002-07-03

    The intensive use of antimicrobial agents, mainly oxytetracycline, to prevent and control bacterial pathologies in Chilean salmon culture is a frequent practice. A total of 103 gram-negative oxytetracycline-resistant bacteria recovered from various sources of 4 Chilean freshwater salmon farms were identified and investigated for their susceptibility patterns to various antibacterial agents, by using an agar disk diffusion method. Antibacterial resistance patterns of isolates were not correlated with bacterial species or strain source. A high number of bacteria resistant to amoxicillin, ampicillin. erythromycin, and furazolidone, as well as an important frequency of bacterial resistance to florfenicol, chloramphenicol, cefotaxime and trimethoprim-sulfamethoxazole was found. On the contrary, the proportion of bacteria resistant to gentamicin, kanamycin, flumequine and enrofloxacin was rather low. Resistant microflora showed a high taxonomic variability and mainly consisted of non-fermenting bacteria (77.7%). These strains mainly belonged to the species Pseudomonas fluorescens (29), Aeromonas hydrophila (10), Stenotrophomonas maltophilia (6), isolated from salmon fingerlings, and Acinetobacter lwoffii (5) isolated from pelletized feed. The occurrence of simultaneous resistance to various antibacterials was frequent. We observe a high frequency of bacteria resistant to 6-10 antibacterials (74 strains), and antibiotic resistance index (ARI) values ranging from 0.38 to 0.48 for the four salmon farms studied. These results suggest that Chilean salmon farms might play a role as reservoirs of antibacterial multiresistant bacteria, thus prompting the necessity for a more restrictive attitude towards the intensive use of antibacterials in salmon farming.

  15. Pseudomonas aeruginosa in Musca domestica L.: temporospatial examination of bacteria population dynamics and house fly antimicrobial responses

    Science.gov (United States)

    House flies associate with microbes throughout their life history. Bacteria ingested by adult flies enter the alimentary canal and face a hostile environment including antimicrobial defenses. Because the outcome of this interaction impacts bacterial survival and dissemination, our primary objective ...

  16. 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 ...

  17. Microcystis aeruginosa/Pseudomonas pseudoalcaligenes interaction effects on off-flavors in algae/bacteria co-culture system under different temperatures.

    Science.gov (United States)

    Yang, Xi; Xie, Ping; Yu, Yunzhen; Shen, Hong; Deng, Xuwei; Ma, Zhimei; Wang, Peili; Tao, Min; Niu, Yuan

    2015-05-01

    We conducted an experiment to study the interaction effects of Microcystis aeruginosa and Pseudomonas pseudoalcaligenes on off-flavors in an algae/bacteria co-culture system at three temperatures (24, 28 and 32°C). Gas chromatography-mass spectrometry was applied to measure off-flavor compounds dimethyl sulfide (DMS), dimethyl trisulfide (DMTS), 2-methylisoborneol, geosmin (GEO) and β-cyclocitral. During the lag phase of co-cultured M. aeruginosa (first 15days), P. pseudoalcaligenes significantly increased the production of DMS, DMTS and β-cyclocitral at all three temperatures. In the exponential phase of co-cultured M. aeruginosa (after 15days), M. aeruginosa became the main factor on off-flavors in the co-culture system, and β-cyclocitral turned to the highest off-flavor compound. These results also indicated that DMS, DMTS and β-cyclocitral were the main off-flavor compounds in our M. aeruginosa/P. pseudoalcaligenes co-culture system. Univariate analysis was applied to investigate the effects of M. aeruginosa and P. pseudoalcaligenes on the production of off-flavors. The results demonstrated that both M. aeruginosa and P. pseudoalcaligenes could increase the production of DMS and DMTS, while β-cyclocitral was mainly determined by M. aeruginosa. Our results also provide some insights into understanding the relationship between cyanobacteria and heterotrophic bacteria. Copyright © 2015. Published by Elsevier B.V.

  18. Bacteria associated with Amblyomma cajennense tick eggs

    Directory of Open Access Journals (Sweden)

    Erik Machado-Ferreira

    2015-01-01

    Full Text Available AbstractTicks represent a large group of pathogen vectors that blood feed on a diversity of hosts. In the Americas, the Ixodidae ticks Amblyomma cajennense are responsible for severe impact on livestock and public health. In the present work, we present the isolation and molecular identification of a group of culturable bacteria associated with A. cajennense eggs from females sampled in distinct geographical sites in southeastern Brazil. Additional comparative analysis of the culturable bacteria from Anocentor nitens, Rhipicephalus sanguineus and Ixodes scapularis tick eggs were also performed. 16S rRNA gene sequence analyses identified 17 different bacterial types identified as Serratia marcescens, Stenotrophomonas maltophilia, Pseudomonas fluorescens, Enterobacter spp., Micrococcus luteus, Ochrobactrum anthropi, Bacillus cereus and Staphylococcus spp., distributed in 12 phylogroups. Staphylococcus spp., especially S. sciuri,was the most prevalent bacteria associated with A. cajennenseeggs, occurring in 65% of the samples and also frequently observed infecting A. nitens eggs. S. maltophilia, S. marcescens and B. cereus occurred infecting eggs derived from specific sampling sites, but in all cases rising almost as pure cultures from infected A. cajennense eggs. The potential role of these bacterial associations is discussed and they possibly represent new targets for biological control strategies of ticks and tick borne diseases.

  19. Study in vitro of the impact of endophytic bacteria isolated from Centella asiatica on the disease incidence caused by the hemibiotrophic fungus Colletotrichum higginsianum.

    Science.gov (United States)

    Rakotoniriana, Erick Francisco; Rafamantanana, Mamy; Randriamampionona, Denis; Rabemanantsoa, Christian; Urveg-Ratsimamanga, Suzanne; El Jaziri, Mondher; Munaut, Françoise; Corbisier, Anne-Marie; Quetin-Leclercq, Joëlle; Declerck, Stéphane

    2013-01-01

    Thirty-one endophytic bacteria isolated from healthy leaves of Centella asiatica were screened in vitro for their ability to reduce the growth rate and disease incidence of Colletotrichum higginsianum, a causal agent of anthracnose. Isolates of Cohnella sp., Paenibacillus sp. and Pantoea sp. significantly stimulated the growth rate of C. higginsianum MUCL 44942, while isolates of Achromobacter sp., Acinetobacter sp., Microbacterium sp., Klebsiella sp. and Pseudomonas putida had no influence on this plant pathogen. By contrast, Bacillus subtilis BCA31 and Pseudomonas fluorescens BCA08 caused a marked inhibition of C. higginsianum MUCL 44942 growth by 46 and 82 %, respectively. Cell-free culture filtrates of B. subtilis BCA31 and P. fluorescens BCA08 were found to contain antifungal compounds against C. higginsianum MUCL 44942. Inoculation assays on in vitro-cultured plants of C. asiatica showed that foliar application of B. subtilis BCA31, three days before inoculation with C. higginsianum MUCL 44942, significantly reduced incidence and severity of the disease. The role of endophytic bacteria in maintaining the apparent inactivity of C. higginsianum MUCL 44942 in C. asiatica grown in the wild is discussed.

  20. Escherichia coli BdcA controls biofilm dispersal in Pseudomonas aeruginosa and Rhizobium meliloti

    Directory of Open Access Journals (Sweden)

    Wood Thomas K

    2011-10-01

    Full Text Available Abstract Background Previously we showed that BdcA controls Escherichia coli biofilm dispersal by binding the ubiquitous bacterial signal cyclic diguanylate (c-di-GMP; upon reducing the concentration of c-di-GMP, the cell shifts to the planktonic state by increasing motility, decreasing aggregation, and decreasing production of biofilm adhesins. Findings Here we report that BdcA also increases biofilm dispersal in other Gram-negative bacteria including Pseudomonas aeruginosa, Pseudomonas fluorescens, and Rhizobium meliloti. BdcA binds c-di-GMP in these strains and thereby reduces the effective c-di-GMP concentrations as demonstrated by increases in swimming motility and swarming motility as well as by a reduction in extracellular polysaccharide production. We also develop a method to displace existing biofilms by adding BdcA via conjugation from E. coli in mixed-species biofilms. Conclusion Since BdcA shows the ability to control biofilm dispersal in diverse bacteria, BdcA has the potential to be used as a tool to disperse biofilms for engineering and medical applications.

  1. Penicillin V acylases from gram-negative bacteria degrade N-acylhomoserine lactones and attenuate virulence in Pseudomonas aeruginosa

    NARCIS (Netherlands)

    Sunder, Avinash Vellore; Utari, Putri Dwi; Ramasamy, Sureshkumar; van Merkerk, Ronald; Quax, Wim J.; Pundle, Archana

    Virulence pathways in gram-negative pathogenic bacteria are regulated by quorum sensing mechanisms, through the production and sensing of N-acylhomoserine lactone (AHL) signal molecules. Enzymatic degradation of AHLs leading to attenuation of virulence (quorum quenching) could pave the way for the

  2. Differential sensitivity of polyhydroxyalkanoate producing bacteria to fermentation inhibitors and comparison of polyhydroxybutyrate production from Burkholderia cepacia and Pseudomonas pseudoflava

    Science.gov (United States)

    Diane Dietrich; Barbara Illman; Casey Crooks

    2013-01-01

    The aim of this study is determine the relative sensitivity of a panel of seven polyhydroxyalkanoate producing bacteria to a panel of seven lignocellulosic-derived fermentation inhibitors representing aliphatic acids, furans and phenolics. A further aim was to measure the polyhydroxybutyrate production of select organisms on lignocellulosic-derived monosaccharides...

  3. Results from a 13-Year Prospective Cohort Study Show Increased Mortality Associated with Bloodstream Infections Caused by Pseudomonas aeruginosa Compared to Other Bacteria.

    Science.gov (United States)

    Thaden, Joshua T; Park, Lawrence P; Maskarinec, Stacey A; Ruffin, Felicia; Fowler, Vance G; van Duin, David

    2017-06-01

    The impact of bacterial species on outcome in bloodstream infections (BSI) is incompletely understood. We evaluated the impact of bacterial species on BSI mortality, with adjustment for patient, bacterial, and treatment factors. From 2002 to 2015, all adult inpatients with monomicrobial BSI caused by Staphylococcus aureus or Gram-negative bacteria at Duke University Medical Center were prospectively enrolled. Kaplan-Meier curves and multivariable Cox regression with propensity score models were used to examine species-specific bacterial BSI mortality. Of the 2,659 enrolled patients, 999 (38%) were infected with S. aureus, and 1,660 (62%) were infected with Gram-negative bacteria. Among patients with Gram-negative BSI, Enterobacteriaceae (81% [1,343/1,660]) were most commonly isolated, followed by non-lactose-fermenting Gram-negative bacteria (16% [262/1,660]). Of the 999 S. aureus BSI isolates, 507 (51%) were methicillin resistant. Of the 1,660 Gram-negative BSI isolates, 500 (30%) were multidrug resistant. The unadjusted time-to-mortality among patients with Gram-negative BSI was shorter than that of patients with S. aureus BSI (P = 0.003), due to increased mortality in patients with non-lactose-fermenting Gram-negative BSI generally (P < 0.0001) and Pseudomonas aeruginosa BSI (n = 158) in particular (P < 0.0001). After adjustment for patient demographics, medical comorbidities, bacterial antibiotic resistance, timing of appropriate antibiotic therapy, and source control in patients with line-associated BSI, P. aeruginosa BSI remained significantly associated with increased mortality (hazard ratio = 1.435; 95% confidence interval = 1.043 to 1.933; P = 0.02). P. aeruginosa BSI was associated with increased mortality relative to S. aureus or other Gram-negative BSI. This effect persisted after adjustment for patient, bacterial, and treatment factors. Copyright © 2017 American Society for Microbiology.

  4. Role of the lysozyme inhibitor Ivy in growth or survival of Escherichia coli and Pseudomonas aeruginosa bacteria in hen egg white and in human saliva and breast milk.

    Science.gov (United States)

    Deckers, Daphne; Vanlint, Dietrich; Callewaert, Lien; Aertsen, Abram; Michiels, Chris W

    2008-07-01

    Ivy is a lysozyme inhibitor that protects Escherichia coli against lysozyme-mediated cell wall hydrolysis when the outer membrane is permeabilized by mutation or by chemical or physical stress. In the current work, we have investigated whether Ivy is necessary for the survival or growth of E. coli MG1655 and Pseudomonas aeruginosa PAO1 in hen egg white and in human saliva and breast milk, which are naturally rich in lysozyme and in membrane-permeabilizing components. Wild-type E. coli was able to grow in saliva and breast milk but showed partial inactivation in egg white. The knockout of Ivy did not affect growth in breast milk but slightly increased sensitivity to egg white and caused hypersensitivity to saliva, resulting in the complete inactivation of 10(4) CFU ml(-1) of bacteria within less than 5 hours. The depletion of lysozyme from saliva completely restored the ability of the ivy mutant to grow like the parental strain. P. aeruginosa, in contrast, showed growth in all three substrates, which was not affected by the knockout of Ivy production. These results indicate that lysozyme inhibitors like Ivy promote bacterial survival or growth in particular lysozyme-rich secretions and suggest that they may promote the bacterial colonization of specific niches in the animal host.

  5. Isolation and characterization of biosurfactant producing bacteria from Persian Gulf (Bushehr provenance).

    Science.gov (United States)

    Hassanshahian, Mehdi

    2014-09-15

    Biosurfactants are surface active materials that are produced by some microorganisms. These molecules increase biodegradation of insoluble pollutants. In this study sediments and seawater samples were collected from the coastline of Bushehr provenance in the Persian Gulf and their biosurfactant producing bacteria were isolated. Biosurfactant producing bacteria were isolated by using an enrichment method in Bushnell-Hass medium with diesel oil as the sole carbon source. Five screening tests were used for selection of Biosurfactant producing bacteria: hemolysis in blood agar, oil spreading, drop collapse, emulsification activity and Bacterial Adhesion to Hydrocarbon test (BATH). These bacteria were identified using biochemical and molecular methods. Eighty different colonies were isolated from the collected samples. The most biosurfactant producing isolates related to petrochemical plants of Khark Island. Fourteen biosurfactant producing bacteria were selected between these isolates and 7 isolates were screened as these were predominant producers that belong to Shewanella alga, Shewanella upenei, Vibrio furnissii, Gallaecimonas pentaromativorans, Brevibacterium epidermidis, Psychrobacter namhaensis and Pseudomonas fluorescens. The largest clear zone diameters in oil spreading were observed for G. pentaromativorans strain O15. Also, this strain has the best emulsification activity and reduction of surface tension, suggesting it is the best of thee isolated strains. The results of this study confirmed that there is high diversity of biosurfactant producing bacteria in marine ecosystem of Iran and by application of these bacteria in petrochemical waste water environmental problems can be assisted. Copyright © 2014 Elsevier Ltd. All rights reserved.

  6. Bacteria transport and retention in intact calcareous soil columns under saturated flow conditions

    Directory of Open Access Journals (Sweden)

    Farrokhian Firouzi Ahmad

    2015-06-01

    Full Text Available Study of bacterial transport and retention in soil is important for various environmental applications such as groundwater contamination and bioremediation of soil and water. The main objective of this research was to quantitatively assess bacterial transport and deposition under saturated conditions in calcareous soil. A series of leaching experiments was conducted on two undisturbed soil columns. Breakthrough curves of Pseudomonas fluorescens and Cl were measured. After the leaching experiment, spatial distribution of bacteria retention in the soil columns was determined. The HYDRUS-1D one- and two-site kinetic models were used to predict the transport and deposition of bacteria in soil. The results indicated that the two-site model fits the observed data better than one-site kinetic model. Bacteria interaction with the soil of kinetic site 1 revealed relatively fast attachment and slow detachment, whereas attachment to and detachment of bacteria from kinetic site 2 was fast. Fast attachment and slow detachment of site 1 can be attributed to soil calcium carbonate that has favorable attachment sites for bacteria. The detachment rate was less than 0.02 of the attachment rate, indicating irreversible attachment of bacteria. High reduction rate of bacteria was also attributed to soil calcium carbonate.

  7. Antibacterial activity of bark extracts of Moringa oleifera Lam. against some selected bacteria.

    Science.gov (United States)

    Zaffer, Mudasser; Ahmad, Showkat; Sharma, Rajendra; Mahajan, Surabhi; Gupta, Ankur; Agnihotri, Rajneesh Kumar

    2014-11-01

    The methanol, chloroform, ethyl acetate and aqueous bark extracts of Moringa oleifera were evaluated for their antibacterial activity against four bacteria viz. Staphylococcus aureus, Citrobacter freundii, Bacillus megaterium and Pseudomonas fluorescens using erythromycin as positive control. The activity was analyzed using paper disc diffusion method at different concentration of the extract. The study revealed that all the bark extracts irrespective of their types, in different concentrations inhibited growth of the test pathogens to varying degrees. Ethyl acetate extract showed maximum activity against all the bacterial strains followed in descending order by chloroform, methanol and aqueous extracts. The activity decreased with decrease in concentration of the extract. Staphylococcus aureus was found to be the most sensitive test organism to different extracts of Moringa oleifera. Looking to these results it may be concluded that M. oleifera may be a potential source for the treatment of different infections caused by the resistant microbes.

  8. Investigation of biotechnological potential of sponge-associated bacteria collected in Brazilian coast.

    Science.gov (United States)

    Santos, O C S; Soares, A R; Machado, F L S; Romanos, M T V; Muricy, G; Giambiagi-deMarval, M; Laport, M S

    2015-02-01

    Marine bacteria are a rich source of structurally unique natural compounds, several of which have shown a wide variety of biological activities. In this study, the metabolites present in the culture supernatants of the eight sponge-associated bacteria were extracted using ethyl acetate, and all extracts showed activity against Staphylococcus aureus. Subsequently, the extracts of the Pseudomonas fluorescens H40 and H41, and Pseudomonas aeruginosa H51 were subjected to solvent partitioning, and the active fractions were submitted to chromatographic separation. Three different active fractions were obtained, one of which was identified as diketopiperazine cyclo-(L-Leu-L-Pro). This substance was bactericidal for Staph. aureus and Ps. aeruginosa and showed cytotoxic activity against HEp-2 tumour cells. Putative gene fragments coding for the type I polyketide synthase (PKS-I) and nonribosomal peptide synthetase (NRPS) domains were PCR-amplified from five and three strains, respectively. The results suggest that sponge-associated bacteria analysed in this study may represent a potential source for production of antimicrobial substances against bacterial pathogens of medical importance. © 2014 The Society for Applied Microbiology.

  9. Interaction of protamine with gram-negative bacteria membranes: possible alternative mechanisms of internalization in Escherichia coli, Salmonella typhimurium and Pseudomonas aeruginosa.

    Science.gov (United States)

    Pink, David A; Hasan, Fida M; Quinn, Bonnie E; Winterhalter, Mathias; Mohan, Mukund; Gill, Tom A

    2014-04-01

    This study was concerned with the interaction between the cationic antimicrobial peptide, protamine (Ptm) and the cytoplasmic membranes of the gram-negative bacteria Escherichia coli, Salmonella typhimurium and Pseudomonas aeruginosa. The objective of the study was to explain the observed paradox of internalization without permanent disruption of the cell envelope. We carried out Monte Carlo computer simulation of Ptm in an aqueous environment in the presence of ~100 mM NaCl and model membranes consisting of either (65:35) or (75:25) PE:PG molar ratios. The (75:25) model, representative of the gram-negative cytoplasmic membrane, showed that the Ptm center of mass remained at least 7 nm from the membrane surface leading to the prediction that Ptm would not internalize via disruption of the inner membrane. By using immunoelectron microscopy of Ptm-treated cells, we showed that Ptm internalization to the cytoplasm took place rapidly in the presence or absence of the outer envelope. Ultrastructural examination revealed no obvious morphological changes to cells that were treated with subinhibitory or bactericidal levels of Ptm. Reconstituted phospholipid bilayers were constructed and were unperturbed by Ptm treatment over a wide range of concentrations and applied transmembrane voltages. We conclude that in the cases of the cell envelopes of E. coli, S. typhimurium and P. aeruginosa, Ptm internalized by means independent of the phospholipid bilayer, most likely mediated by one or more membrane proteins such as cation-selective barrel-like proteins. Work is currently underway to test this hypothesis. © 2014 The Authors. Journal of Peptide Science published by John Wiley & Sons, Ltd.

  10. 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.

  11. 40 CFR 180.1107 - Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Delta endotoxin of Bacillus... From Tolerances § 180.1107 Delta endotoxin of Bacillus thuringiensis variety kurstaki encapsulated into killed Pseudomonas fluorescens; exemption from the requirement of a tolerance. The delta endotoxin of...

  12. The dynamics of apoplast phenolics in tobacco leaves following inoculation with bacteria

    Directory of Open Access Journals (Sweden)

    Con Jacyn Baker

    2015-08-01

    Full Text Available This study demonstrates that the accumulation of apoplastic phenolics is stimulated in planta in response to bacterial inoculation. Past studies have shown that levels of extracellular phenolics are elicited in plant cell suspensions in response to bacteria, and that tomato plants infected with viroids showed changes in apoplastic phenolics. The method described here monitored changes in apoplastic phenolics in tobacco leaves following bacterial inoculation of the same tissue. Inoculation with a saprophyte, Pseudomonas fluorescens, which does not cause visible symptoms or physical damage, was used to elicit phenolics and examine the effects of variable parameters on phenolic composition. Location of the inoculation on the leaf, position or developmental age of the leaf on the plant, and inoculum concentration were standardized for further experiments. The patterns of phenolic change in the apoplast were compared for tobacco inoculated with P. syringae pathovars, pv. syringae, which causes a resistant HR reaction within 15 h, and pv. tabaci, which causes a susceptible reaction with delayed visible symptoms. Both pathogens elicited lower increased levels of acetosyringone compared to the saprophyte, P. fluorescens but had greatly increased levels of the chlorogenic acid derivatives. The latter metabolites appear to have come from the intracellular stores, which could

  13. The dynamics of apoplast phenolics in tobacco leaves following inoculation with bacteria.

    Science.gov (United States)

    Baker, Con J; Mock, Norton M; Smith, Jodi M; Aver'yanov, Andrey A

    2015-01-01

    This study demonstrates that the accumulation of apoplastic phenolics is stimulated in planta in response to bacterial inoculation. Past studies have shown that levels of extracellular phenolics are elicited in plant cell suspensions in response to bacteria, and that tomato plants infected with viroids showed changes in apoplastic phenolics. The method described here monitored changes in apoplastic phenolics in tobacco leaves following bacterial inoculation of the same tissue. Inoculation with a saprophyte, Pseudomonas fluorescens, which does not cause visible symptoms or physical damage, was used to elicit phenolics and examine the effects of variable parameters on phenolic composition. Location of the inoculation on the leaf, position, or developmental age of the leaf on the plant, and inoculum concentration were standardized for further experiments. The patterns of phenolic change in the apoplast were compared for tobacco inoculated with P. syringae pathovars, pv. syringae, which causes a resistant HR reaction within 15 h, and pv. tabaci, which causes a susceptible reaction with delayed visible symptoms. Both pathogens elicited lower increased levels of acetosyringone compared to the saprophyte, P. fluorescens but had greatly increased levels of the chlorogenic acid derivatives. The latter metabolites appear to have come from the intracellular stores, which could indicate a weakening of the apoplast/symplast barrier. This unexpected aspect will require further study of intracellular phenolics.

  14. Identification of flgZ as a flagellar gene encoding a PilZ domain protein that regulates swimming motility and biofilm formation in Pseudomonas.

    Directory of Open Access Journals (Sweden)

    Francisco Martínez-Granero

    Full Text Available Diguanylate cyclase and phosphodiesterase enzymatic activities control c-di-GMP levels modulating planktonic versus sessile lifestyle behavior in bacteria. The PilZ domain is described as a sensor of c-di-GMP intracellular levels and the proteins containing a PilZ domain represent the best studied class of c-di-GMP receptors forming part of the c-di-GMP signaling cascade. In P. fluorescens F113 we have found two diguanylate cyclases (WspR, SadC and one phosphodiesterase (BifA implicated in regulation of swimming motility and biofilm formation. Here we identify a flgZ gene located in a flagellar operon encoding a protein that contains a PilZ domain. Moreover, we show that FlgZ subcellular localization depends on the c-di-GMP intracellular levels. The overexpression analysis of flgZ in P. fluorescens F113 and P. putida KT2440 backgrounds reveal a participation of FlgZ in Pseudomonas swimming motility regulation. Besides, the epistasis of flgZ over wspR and bifA clearly shows that c-di-GMP intracellular levels produced by the enzymatic activity of the diguanylate cyclase WspR and the phosphodiesterase BifA regulates biofilm formation through FlgZ.

  15. Contribution of increased mutagenesis to the evolution of pollutants-degrading indigenous bacteria.

    Science.gov (United States)

    Ilmjärv, Tanel; Naanuri, Eve; Kivisaar, Maia

    2017-01-01

    Bacteria can rapidly evolve mechanisms allowing them to use toxic environmental pollutants as a carbon source. In the current study we examined whether the survival and evolution of indigenous bacteria with the capacity to degrade organic pollutants could be connected with increased mutation frequency. The presence of constitutive and transient mutators was monitored among 53 pollutants-degrading indigenous bacterial strains. Only two strains expressed a moderate mutator phenotype and six were hypomutators, which implies that constitutively increased mutability has not been prevalent in the evolution of pollutants degrading bacteria. At the same time, a large proportion of the studied indigenous strains exhibited UV-irradiation-induced mutagenesis, indicating that these strains possess error-prone DNA polymerases which could elevate mutation frequency transiently under the conditions of DNA damage. A closer inspection of two Pseudomonas fluorescens strains PC20 and PC24 revealed that they harbour genes for ImuC (DnaE2) and more than one copy of genes for Pol V. Our results also revealed that availability of other nutrients in addition to aromatic pollutants in the growth environment of bacteria affects mutagenic effects of aromatic compounds. These results also implied that mutagenicity might be affected by a factor of how long bacteria have evolved to use a particular pollutant as a carbon source.

  16. Contribution of increased mutagenesis to the evolution of pollutants-degrading indigenous bacteria.

    Directory of Open Access Journals (Sweden)

    Tanel Ilmjärv

    Full Text Available Bacteria can rapidly evolve mechanisms allowing them to use toxic environmental pollutants as a carbon source. In the current study we examined whether the survival and evolution of indigenous bacteria with the capacity to degrade organic pollutants could be connected with increased mutation frequency. The presence of constitutive and transient mutators was monitored among 53 pollutants-degrading indigenous bacterial strains. Only two strains expressed a moderate mutator phenotype and six were hypomutators, which implies that constitutively increased mutability has not been prevalent in the evolution of pollutants degrading bacteria. At the same time, a large proportion of the studied indigenous strains exhibited UV-irradiation-induced mutagenesis, indicating that these strains possess error-prone DNA polymerases which could elevate mutation frequency transiently under the conditions of DNA damage. A closer inspection of two Pseudomonas fluorescens strains PC20 and PC24 revealed that they harbour genes for ImuC (DnaE2 and more than one copy of genes for Pol V. Our results also revealed that availability of other nutrients in addition to aromatic pollutants in the growth environment of bacteria affects mutagenic effects of aromatic compounds. These results also implied that mutagenicity might be affected by a factor of how long bacteria have evolved to use a particular pollutant as a carbon source.

  17. The Pseudomonas aeruginosa oxyvinylglycine L-2-amino-4-methoxy-trans-3-butenoic acid inhibits growth of Erwinia amylovora and acts as a weak seed germination-arrest factor

    Science.gov (United States)

    The Pseudomonas aeruginosa antimetabolite L-2-amino-4-methoxy-trans-3-butenoic acid (AMB) is demonstrated to share biological activities with 4-formylaminooxyvinylglycine, a related molecule produced by Pseudomonas fluorescens WH6. We found that culture filtrates of a P. aeruginosa strain overproduc...

  18. Isolation and characterization of bacteria from the gut of Bombyx mori that degrade cellulose, xylan, pectin and starch and their impact on digestion.

    Science.gov (United States)

    Anand, A Alwin Prem; Vennison, S John; Sankar, S Gowri; Prabhu, D Immanual Gilwax; Vasan, P Thirumalai; Raghuraman, T; Geoffrey, C Jerome; Vendan, S Ezhil

    2010-01-01

    Bombyx mori L. (Lepidoptera: Bombycidae) have been domesticated and widely used for silk production. It feeds on mulberry leaves. Mulberry leaves are mainly composed of pectin, xylan, cellulose and starch. Some of the digestive enzymes that degrade these carbohydrates might be produced by gut bacteria. Eleven isolates were obtained from the digestive tract of B. mori, including the Gram positive Bacillus circulans and Gram negative Proteus vulgaris, Klebsiella pneumoniae, Escherichia coli, Citrobacter freundii, Serratia liquefaciens, Enterobacter sp., Pseudomonas fluorescens, P. aeruginosa, Aeromonas sp., and Erwinia sp.. Three of these isolates, P. vulgaris, K. pneumoniae, C. freundii, were cellulolytic and xylanolytic, P. fluorescens and Erwinia sp., were pectinolytic and K. pneumoniae degraded starch. Aeromonas sp. was able to utilize the CMcellulose and xylan. S. liquefaciens was able to utilize three polysaccharides including CMcellulose, xylan and pectin. B. circulans was able to utilize all four polysaccharides with different efficacy. The gut of B. mori has an alkaline pH and all of the isolated bacterial strains were found to grow and degrade polysaccharides at alkaline pH. The number of cellulolytic bacteria increases with each instar.

  19. Isolation and Characterization of Bacteria from the Gut of Bombyx mori that Degrade Cellulose, Xylan, Pectin and Starch and Their Impact on Digestion

    Science.gov (United States)

    Anand, A. Alwin Prem; Vennison, S. John; Sankar, S. Gowri; Prabhu, D. Immanual Gilwax; Vasan, P. Thirumalai; Raghuraman, T.; Geoffrey, C. Jerome; Vendan, S. Ezhil

    2010-01-01

    Bombyx mori L. (Lepidoptera: Bombycidae) have been domesticated and widely used for silk production. It feeds on mulberry leaves. Mulberry leaves are mainly composed of pectin, xylan, cellulose and starch. Some of the digestive enzymes that degrade these carbohydrates might be produced by gut bacteria. Eleven isolates were obtained from the digestive tract of B. mori, including the Gram positive Bacillus circulans and Gram negative Proteus vulgaris, Klebsiella pneumoniae, Escherichia coli, Citrobacter freundii, Serratia liquefaciens, Enterobacter sp., Pseudomonas fluorescens, P. aeruginosa, Aeromonas sp., and Erwinia sp.. Three of these isolates, P. vulgaris, K. pneumoniae, C. freundii, were cellulolytic and xylanolytic, P. fluorescens and Erwinia sp., were pectinolytic and K. pneumoniae degraded starch. Aeromonas sp. was able to utilize the CMcellulose and xylan. S. liquefaciens was able to utilize three polysaccharides including CMcellulose, xylan and pectin. B. circulans was able to utilize all four polysaccharides with different efficacy. The gut of B. mori has an alkaline pH and all of the isolated bacterial strains were found to grow and degrade polysaccharides at alkaline pH. The number of cellulolytic bacteria increases with each instar. PMID:20874394

  20. Quorum quenching activity in cell-free lysate of endophytic bacteria isolated from Pterocarpus santalinus Linn., and its effect on quorum sensing regulated biofilm in Pseudomonas aeruginosa PAO1.

    Science.gov (United States)

    Rajesh, P S; Ravishankar Rai, V

    2014-01-01

    Quorum sensing mechanism allows the microorganisms to resist the antibiotic treatment by forming biofilms. Quorum quenching is one of the mechanisms to control the development of drug resistance in microbes. Endophyte bacteria are beneficial to plant growth as they support the immune system against the pathogen attack. The endophytic bacteria present in Pterocarpus santalinus were screened for the presence of N-acyl homoserine lactones (AHLs) degrading bacteria using biosensor strains and further confirmed by quantifying the violacein production. Cell-free lysate of endophytic bacteria, Bacillus firmus PT18 and Enterobacter asburiae PT39 exhibited potent AHL degrading ability by inhibiting about 80% violacein production in biosensor strain. Furthermore, when the cell-free lysate was applied to Pseudomonas aeruginosa PAO1 and PAO1-JP2 biofilm it resulted in significant (p<0.01) inhibition of biofilm formation. The biofilm inhibition was confirmed by visualization of biofilm slides under fluorescence microscopy, which showed decrease in total biomass formation in treated slides. Isolation and amplification of the gene (aiiA) indicated that the presence of AHL lactonase in cell-free lysate and sequence alignment indicated that AiiA contains a "HXHXDH" zinc-binding motif that is being conserved in several groups of metallohydrolases. Therefore, the study shows the potential of AHLs degradation by AHL lactonase present in cell-free lysate of isolated endophytic bacteria and inhibition of quorum sensing regulated biofilm formation in P. aeruginosa PAO1. Copyright © 2013 Elsevier GmbH. All rights reserved.

  1. Protozoan-Induced Regulation of Cyclic Lipopeptide Biosynthesis Is an Effective Predation Defense Mechanism for Pseudomonas fluorescens▿

    Science.gov (United States)

    Mazzola, Mark; de Bruijn, Irene; Cohen, Michael F.; Raaijmakers, Jos M.

    2009-01-01

    Environmental bacteria are exposed to a myriad of biotic interactions that influence their function and survival. The grazing activity of protozoan predators significantly impacts the dynamics, diversification, and evolution of bacterial communities in soil ecosystems. To evade protozoan predation, bacteria employ various defense strategies. Soil-dwelling Pseudomonas fluorescens strains SS101 and SBW25 produce the cyclic lipopeptide surfactants (CLPs) massetolide and viscosin, respectively, in a quorum-sensing-independent manner. In this study, CLP production was shown to protect these bacteria from protozoan predation as, compared to CLP-deficient mutants, strains SS101 and SBW25 exhibited resistance to grazing by Naegleria americana in vitro and superior persistence in soil in the presence of this bacterial predator. In the wheat rhizosphere, CLP-producing strains had a direct deleterious impact on the survival of N. americana. In vitro assays further showed that N. americana was three times more sensitive to viscosin than to massetolide and that exposure of strain SS101 or SBW25 to this protozoan resulted in upregulation of CLP biosynthesis genes. Enhanced expression of the massABC and viscABC genes did not require physical contact between the two organisms as gene expression levels were up to threefold higher in bacterial cells harvested 1 cm from feeding protozoans than in cells collected 4 cm from feeding protozoans. These findings document a new natural function of CLPs and highlight that bacterium-protozoan interactions can result in activation of an antipredator response in prey populations. PMID:19717630

  2. Bactericidal efficacy of elevated pH on fish pathogenic and environmental bacteria

    Directory of Open Access Journals (Sweden)

    Clifford E. Starliper

    2013-07-01

    Full Text Available Ship ballast water is a recognized medium for transfer and introductions of nonindigenous species. There is a need for new ballast water treatment methods that effectively and safely eliminate or greatly minimize movements of these species. The present study employed laboratory methods to evaluate the bactericidal efficacy of increased pH (pH 10.0–12.0 for exposure durations of up to 72 h to kill a variety of Gram-negative and Gram-positive bacteria including fish pathogens (Aeromonas spp., Yersinia ruckeri, Edwardsiella ictaluri, Serratia liquefaciens, Carnobacterium sp., other common aquatic-inhabitant bacteria (Serratia marcescens, Pseudomonas fluorescens, Staphylococcus sp., Bacillus sp. and indicators listed in International Maritime Organization D2 Standards; namely, Vibrio cholera (an environmental isolate from fish, Escherichia coli and Enterococcus faecalis. Volumes of 5 N NaOH were added to tryptic soy broth to obtain desired pH adjustments. Viable cells were determined after 0, 4, 12, 24, 48, and 72 h. Initial (0 h cell numbers ranged from 3.40 × 104 cfu/mL for Bacillus sp. to 2.44 × 107 cfu/mL for E. faecalis. The effective endpoints of pH and treatment duration necessary to realize 100% bactericidal effect varied; however, all bacteria tested were killed within 72 h at pH 12.0 or lower. The lowest parameters examined, 4 h at pH 10.0, were bactericidal to V. cholera, E. ictaluri, three of four isolates of E. coli, and (three of four Aeromonas salmonicida subsp. salmonicida. Bactericidal effect was attained at pH 10.0 within 12 h for the other A. salmonicida subsp. salmonicida, and within 24 h for P. fluorescens, and the remaining E. coli.

  3. Bactericidal efficacy of elevated pH on fish pathogenic and environmental bacteria

    Science.gov (United States)

    Starliper, Clifford E.; Watten, Barnaby J.

    2013-01-01

    Ship ballast water is a recognized medium for transfer and introductions of nonindigenous species. There is a need for new ballast water treatment methods that effectively and safely eliminate or greatly minimize movements of these species. The present study employed laboratory methods to evaluate the bactericidal efficacy of increased pH (pH 10.0–12.0) for exposure durations of up to 72 h to kill a variety of Gram-negative and Gram-positive bacteria including fish pathogens (Aeromonas spp., Yersinia ruckeri, Edwardsiella ictaluri, Serratia liquefaciens, Carnobacterium sp.), other common aquatic-inhabitant bacteria (Serratia marcescens, Pseudomonas fluorescens, Staphylococcus sp., Bacillus sp.) and indicators listed in International Maritime Organization D2 Standards; namely, Vibrio cholera (an environmental isolate from fish), Escherichia coli and Enterococcus faecalis. Volumes of 5 N NaOH were added to tryptic soy broth to obtain desired pH adjustments. Viable cells were determined after 0, 4, 12, 24, 48, and 72 h. Initial (0 h) cell numbers ranged from 3.40 × 104 cfu/mL for Bacillus sp. to 2.44 × 107 cfu/mL for E. faecalis. The effective endpoints of pH and treatment duration necessary to realize 100% bactericidal effect varied; however, all bacteria tested were killed within 72 h at pH 12.0 or lower. The lowest parameters examined, 4 h at pH 10.0, were bactericidal to V. cholera, E. ictaluri, three of four isolates of E. coli, and (three of four) Aeromonas salmonicida subsp. salmonicida. Bactericidal effect was attained at pH 10.0 within 12 h for the other A. salmonicida subsp. salmonicida, and within 24 h for P. fluorescens, and the remaining E. coli.

  4. 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.

    2015-01-01

    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. PMID:26519390

  5. Using phenotype microarrays in the assessment of the antibiotic susceptibility profile of bacteria isolated from wastewater in on-site treatment facilities.

    Science.gov (United States)

    Jałowiecki, Łukasz; Chojniak, Joanna; Dorgeloh, Elmar; Hegedusova, Berta; Ejhed, Helene; Magnér, Jörgen; Płaza, Grażyna

    2017-04-27

    The scope of the study was to apply Phenotype Biolog MicroArray (PM) technology to test the antibiotic sensitivity of the bacterial strains isolated from on-site wastewater treatment facilities. In the first step of the study, the percentage values of resistant bacteria from total heterotrophic bacteria growing on solid media supplemented with various antibiotics were determined. In the untreated wastewater, the average shares of kanamycin-, streptomycin-, and tetracycline-resistant bacteria were 53, 56, and 42%, respectively. Meanwhile, the shares of kanamycin-, streptomycin-, and tetracycline-resistant bacteria in the treated wastewater were 39, 33, and 29%, respectively. To evaluate the antibiotic susceptibility of the bacteria present in the wastewater, using the phenotype microarrays (PMs), the most common isolates from the treated wastewater were chosen: Serratia marcescens ss marcescens, Pseudomonas fluorescens, Stenotrophomonas maltophilia, Stenotrophomonas rhizophila, Microbacterium flavescens, Alcaligenes faecalis ss faecalis, Flavobacterium hydatis, Variovorax paradoxus, Acinetobacter johnsonii, and Aeromonas bestiarum. The strains were classified as multi-antibiotic-resistant bacteria. Most of them were resistant to more than 30 antibiotics from various chemical classes. Phenotype microarrays could be successfully used as an additional tool for evaluation of the multi-antibiotic resistance of environmental bacteria and in preliminary determination of the range of inhibition concentration.

  6. Application of EndophyticPseudomonas fluorescensand a Bacterial Consortium toBrassica napusCan Increase Plant Height and Biomass under Greenhouse and Field Conditions.

    Science.gov (United States)

    Lally, Richard D; Galbally, Paul; Moreira, António S; Spink, John; Ryan, David; Germaine, Kieran J; Dowling, David N

    2017-01-01

    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 Km r 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.

  7. 40 CFR 180.1108 - Delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas...

    Science.gov (United States)

    2010-07-01

    ... From Tolerances § 180.1108 Delta endotoxin of Bacillus thuringiensis variety San Diego encapsulated... of Bacillus thuringiensis variety San Diego encapsulated into killed Pseudomonas fluorescens is... 40 Protection of Environment 23 2010-07-01 2010-07-01 false Delta endotoxin of Bacillus...

  8. 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

  9. Zonulin Regulates Intestinal Permeability and Facilitates Enteric Bacteria Permeation in Coronary Artery Disease.

    Science.gov (United States)

    Li, Chuanwei; Gao, Min; Zhang, Wen; Chen, Caiyu; Zhou, Faying; Hu, Zhangxu; Zeng, Chunyu

    2016-06-29

    Several studies have reported an association between enteric bacteria and atherosclerosis. Bacterial 16S ribosomal RNA (rRNA) gene belong to Enterobacteriaceae have been detected in atherosclerotic plaques. How intestinal bacteria go into blood is not known. Zonulin reversibly modulate intestinal permeability (IP), the circulating zonulin levels were increased in diabetes, obesity, all of which are risk factors for atherosclerosis. It is unclear whether the circulating zonulin levels were changed in coronary artery disease (CAD) patients and modulate IP. The 16S rRNA gene of bacteria in blood sample was checked by 454 pyrosequencing. The zonulin levels were determined by enzyme-linked immunosorbent assay (ELISA) methods. The distribution of zonulin was detected by confocal immunofluorescence microscopy. Bacteria and Caco-2 cell surface micro-structure were checked by transmission electron microscopy. A high diversity of bacterial 16S rRNA gene can be detected in samples from CAD patients, most of them (99.4%) belong to Enterobacteriaceaes, eg. Rahnella. The plasma zonulin levels were significantly higher in CAD patients. Pseudomonas fluorescens exposure significantly increased zonulin expression and decreased IP in a time dependent manner. The elevated zonulin increase IP and may facilitate enteric translocation by disassembling the tight junctions, which might explain the observed high diversity of bacterial 16S rRNA genes in blood samples.

  10. PBAD-based shuttle vectors for functional analysis of toxic and highly regulated genes in Pseudomonas and Burkholderia spp. and other bacteria.

    Science.gov (United States)

    Qiu, Dongru; Damron, F Heath; Mima, Takehiko; Schweizer, Herbert P; Yu, Hongwei D

    2008-12-01

    We report the construction of a series of Escherichia-Pseudomonas broad-host-range expression vectors utilizing the P(BAD) promoter and the araC regulator for routine cloning, conditional expression, and analysis of tightly controlled and/or toxic genes in pseudomonads.

  11. Pseudomonas aeruginosa outcompetes other bacteria in the manifestation and maintenance of a biofilm in polyvinylchloride tubing as used in dental devices.

    Science.gov (United States)

    Ammann, Christoph Gert; Nagl, Markus; Nogler, Michael; Coraça-Huber, Débora Cristina

    2016-05-01

    In a PVC tube as a model system for dental devices, Pseudomonas aeruginosa outcompetes Staphylococcus aureus and Klebsiella pneumoniae for the biofilm formation. P. aeruginosa has advantage over the other strains due to higher tolerance for low-nutrient situations or direct killing by the production of soluble factors like pyocyanin.

  12. Antimicrobial activity of the bioactive components of essential oils from Pakistani spices against Salmonella and other multi-drug resistant bacteria

    Science.gov (United States)

    2013-01-01

    Background The main objective of this study was the phytochemical characterization of four indigenous essential oils obtained from spices and their antibacterial activities against the multidrug resistant clinical and soil isolates prevalent in Pakistan, and ATCC reference strains. Methods Chemical composition of essential oils from four Pakistani spices cumin (Cuminum cyminum), cinnamon (Cinnamomum verum), cardamom (Amomum subulatum) and clove (Syzygium aromaticum) were analyzed on GC/MS. Their antibacterial activities were investigated by minimum inhibitory concentration (MIC) and Thin-Layer Chromatography-Bioautographic (TLC-Bioautographic) assays against pathogenic strains Salmonella typhi (D1 Vi-positive), Salmonella typhi (G7 Vi-negative), Salmonella paratyphi A, Escherichia coli (SS1), Staphylococcus aureus, Pseudomonas fluorescens and Bacillus licheniformis (ATCC 14580). The data were statistically analyzed by using Analysis of Variance (ANOVA) and Least Significant Difference (LSD) method to find out significant relationship of essential oils biological activities at p spices can be pursued against multidrug resistant bacteria. PMID:24119438

  13. 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.

  14. Complete Genome Sequences of Pseudomonas monteilii SB3078 and SB3101, Two Benzene-, Toluene-, and Ethylbenzene-Degrading Bacteria Used for Bioaugmentation

    Science.gov (United States)

    Albertsen, Mads; D’Imperio, Seth; Tale, Vaibhav P.; Lewis, Derrick; Nielsen, Per Halkjær; Nielsen, Jeppe Lund

    2014-01-01

    Pseudomonas monteilii SB3078 and SB3101 are benzene-, toluene-, and ethylbenzene-degrading strains used for bioaugmentation in relation to treatment of wastewater contaminated with petrochemical hydrocarbons. Complete genome sequencing of the bioaugmentation strains confirms that they are very closely related (100.0% average nucleotide identity). Both strains contain extensive integration of phage elements, with the main difference being insertion of additional phage elements in the SB3078 genome. PMID:24874689

  15. Effect of medicinal plants, Heavy metals and antibiotics against pathogenic bacteria isolated from raw, Boiled and pasteurized milk.

    Science.gov (United States)

    Ali, Nazish Mazhar; Sarwar, Khadija; Mazhar, Syed Abdullah; Liaqat, Iram; Andleeb, Saiqa; Mazhar, Bushra; Kalim, Bushra

    2017-11-01

    Present study has been undertaken to isolate and identify the bacterial flora in raw, boiled and pasteurized milk. Agar disc diffusion method was used to determine their sensitivity using medicinal plants, antibiotics and heavy metals. Methylene blue reduction test was used to test the quality of milk samples. Total 10 pathogenic strains were isolated, five strains were isolated from raw milk, three from boiled milk and 2 two from pasteurized milk. To determine optimum conditions for growth, these pathogenic microorganisms were incubated at various temperatures and pH. Gram's staining and biochemical tests revealed that these pathogenic bacteria include Lactobacillus sp., E. coli, Salmonella sp., Pseudomonas sp., Streptococcus sp. and Staphylococcus. Ribotyping revealed S2 as Pseudomonas fluorescens, S5 as Lactococcus lactis and S9 as Lactobacillus acidophilus. Prevalence of pathogenic organisms provided the evidence that contamination of milk arises during milking, transportation and storage of milk. Raw milk is more contaminated than other two types of milk because it contains highest percentage of pathogenic organisms and pasteurized milk was found to be of best quality among three types. So it is recommended to drink milk after proper boiling or pasteurization. Proper pasteurization and hygienic packing of milk is essential to minimize contamination in milk which can save human beings from many milk borne diseases. Our study suggests that antimicrobial use in animal husbandry should be minimized to reduce the hazard of antibiotic resistance. Plant extracts are better alternative against pathogenic bacteria in milk.

  16. Evaluation of the spoilage potential of bacteria isolated from chilled chicken in vitro and in situ.

    Science.gov (United States)

    Wang, Guang-Yu; Wang, Hu-Hu; Han, Yi-Wei; Xing, Tong; Ye, Ke-Ping; Xu, Xing-Lian; Zhou, Guang-Hong

    2017-05-01

    Microorganisms play an important role in the spoilage of chilled chicken. In this study, a total of 53 isolates, belonging to 7 species of 3 genera, were isolated using a selective medium based on the capacity to spoil chicken juice. Four isolates, namely Aeromonas salmonicida 35, Pseudomonas fluorescens H5, Pseudomonas fragi H8 and Serratia liquefaciens 17, were further characterized to assess their proteolytic activities in vitro using meat protein extracts and to evaluate their spoilage potential in situ. The in vitro studies showed that A. salmonicida 35 displayed the strongest proteolytic activity against both sarcoplasmic and myofibrillar proteins. However, the major spoilage isolate in situ was P. fragi H8, which exhibited a fast growth rate, slime formation and increased pH and total volatile basic nitrogen (TVBN) on chicken breast fillets. The relative amounts of volatile organic compounds (VOCs) originating from the microorganisms, including alcohols, aldehydes, ketones and several sulfur compounds, increased during storage. In sum, this study demonstrated the characteristics of 4 potential spoilage bacteria on chilled yellow-feather chicken and provides a simple and convenient method to assess spoilage bacteria during quality management. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. A Proposal for a Genome Similarity-Based Taxonomy for Plant-Pathogenic Bacteria that Is Sufficiently Precise to Reflect Phylogeny, Host Range, and Outbreak Affiliation Applied to Pseudomonas syringae sensu lato as a Proof of Concept.

    Science.gov (United States)

    Vinatzer, Boris A; Weisberg, Alexandra J; Monteil, Caroline L; Elmarakeby, Haitham A; Sheppard, Samuel K; Heath, Lenwood S

    2017-01-01

    Taxonomy of plant pathogenic bacteria is challenging because pathogens of different crops often belong to the same named species but current taxonomy does not provide names for bacteria below the subspecies level. The introduction of the host range-based pathovar system in the 1980s provided a temporary solution to this problem but has many limitations. The affordability of genome sequencing now provides the opportunity for developing a new genome-based taxonomic framework. We already proposed to name individual bacterial isolates based on pairwise genome similarity. Here, we expand on this idea and propose to use genome similarity-based codes, which we now call life identification numbers (LINs), to describe and name bacterial taxa. Using 93 genomes of Pseudomonas syringae sensu lato, LINs were compared with a P. syringae genome tree whereby the assigned LINs were found to be informative of a majority of phylogenetic relationships. LINs also reflected host range and outbreak association for strains of P. syringae pathovar actinidiae, a pathovar for which many genome sequences are available. We conclude that LINs could provide the basis for a new taxonomic framework to address the shortcomings of the current pathovar system and to complement the current taxonomic system of bacteria in general.

  18. Increasing potassium (K release from K-containing minerals in the presence of insoluble phosphate by bacteria

    Directory of Open Access Journals (Sweden)

    Mohammad Reza Sarikhani

    2016-03-01

    Full Text Available Introduction: Phosphorus and potassium are major essential macronutrients for biological growth and development. Application of soil microorganisms is one approach to enhance crop growth. Some bacteria are efficient in releasing K and solubilizing P from mineral sources but their behavior was not studied more in presence together. Materials and methods: In this study the ability of seven bacterial strains, including Pseudomonas putida P13, P. putida Tabriz, P. fluorescens Tabriz, P. fluorescens Chao, Pantoea agglomerans P5, Azotobacter sp. and Bacillus megaterium JK3 to release mineral K from muscovite and biotite with application of insoluble (Ca3(PO42 or soluble (Na2HPO4 P-sources was investigated. Nutrient Broth was used to prepare an overnight culture of bacteria to inoculate in Aleksandrov medium, which was used to study the dissolution of silicate minerals. It should be mentioned that Aleksandrov medium was used to determine the amount of released P from tricalcium phosphate (TCP while muscovite was added to the medium as a sole source of potassium. Concentration of P was determined spectrophotometrically by ammonium-vanadate-molybdate method and K was determined by flame photometry. Results: The insoluble P-source led to a significantly increased released K into assay medium (66%, and the net release of K from the biotite was significantly enhanced. Among bacterial strains, the highest mean of released K was observed with P. putida P13 which released more K (27% than the control. The amounts of released K from micas in the presence of insoluble and soluble phosphate by P. putida P13 were 8.25 and 4.87 mg/g, respectively. Discussion and conclusion: Application of insoluble phosphate could increase K release from mica minerals. The enhanced releasing of mineral K might be attributed to the release of organic acids from the bacteria, a mechanism which plays a pivotal role in solubilizing phosphate from inorganic source of phosphate.

  19. Survey of antibiotic-resistant bacteria isolated from bottlenose dolphins Tursiops truncatus in the southeastern USA.

    Science.gov (United States)

    Stewart, Jill R; Townsend, Forrest I; Lane, Suzanne M; Dyar, Elizabeth; Hohn, Aleta A; Rowles, Teri K; Staggs, Lydia A; Wells, Randall S; Balmer, Brian C; Schwacke, Lori H

    2014-02-19

    Contamination of coastal waters can carry pathogens and contaminants that cause diseases in humans and wildlife, and these pathogens can be transported by water to areas where they are not indigenous. Marine mammals may be indicators of potential health effects from such pathogens and toxins. Here we isolated bacterial species of relevance to humans from wild bottlenose dolphins Tursiops truncatus and assayed isolated bacteria for antibiotic resistance. Samples were collected during capture-release dolphin health assessments at multiple coastal and estuarine sites along the US mid-Atlantic coast and the Gulf of Mexico. These samples were transported on ice and evaluated using commercial systems and aerobic culture techniques routinely employed in clinical laboratories. The most common bacteria identified were species belonging to the genus Vibrio, although Escherichia coli, Shewanella putrefaciens, and Pseudomonas fluorescens/putida were also common. Some of the bacterial species identified have been associated with human illness, including a strain of methicillin-resistant Staphylococcus aureus (MRSA) identified in 1 sample. Widespread antibiotic resistance was observed among all sites, although the percentage of resistant isolates varied across sites and across time. These data provide a baseline for future comparisons of the bacteria that colonize bottlenose dolphins in the southeastern USA.

  20. High Rate of N2 Fixation by East Siberian Cryophilic Soil Bacteria as Determined by Measuring Acetylene Reduction in Nitrogen-Poor Medium Solidified with Gellan Gum▿ †

    Science.gov (United States)

    Hara, Shintaro; Hashidoko, Yasuyuki; Desyatkin, Roman V.; Hatano, Ryusuke; Tahara, Satoshi

    2009-01-01

    For evaluating N2 fixation of diazotrophic bacteria, nitrogen-poor liquid media supplemented with at least 0.5% sugar and 0.2% agar are widely used for acetylene reduction assays. In such a soft gel medium, however, many N2-fixing soil bacteria generally show only trace acetylene reduction activity. Here, we report that use of a N2 fixation medium solidified with gellan gum instead of agar promoted growth of some gellan-preferring soil bacteria. In a soft gel medium solidified with 0.3% gellan gum under appropriate culture conditions, bacterial microbiota from boreal forest bed soils and some free-living N2-fixing soil bacteria isolated from the microbiota exhibited 10- to 200-fold-higher acetylene reduction than those cultured in 0.2% agar medium. To determine the N2 fixation-activating mechanism of gellan gum medium, qualitative differences in the colony-forming bacterial components from tested soil microbiota were investigated in plate cultures solidified with either agar or gellan gum for use with modified Winogradsky's medium. On 1.5% agar plates, apparently cryophilic bacterial microbiota showed strictly distinguishable microbiota according to the depth of soil in samples from an eastern Siberian Taiga forest bed. Some pure cultures of proteobacteria, such as Pseudomonas fluorescens and Burkholderia xenovorans, showed remarkable acetylene reduction. On plates solidified with 1.0% gellan gum, some soil bacteria, including Luteibacter sp., Janthinobacterium sp., Paenibacillus sp., and Arthrobacter sp., uniquely grew that had not grown in the presence of the same inoculants on agar plates. In contrast, Pseudomonas spp. and Burkholderia spp. were apparent only as minor colonies on the gellan gum plates. Moreover, only gellan gum plates allowed some bacteria, particularly those isolated from the shallow organic soil layer, to actively swarm. In consequence, gellan gum is a useful gel matrix to bring out growth potential capabilities of many soil diazotrophs and

  1. High rate of N2 fixation by East Siberian cryophilic soil bacteria as determined by measuring acetylene reduction in nitrogen-poor medium solidified with gellan gum.

    Science.gov (United States)

    Hara, Shintaro; Hashidoko, Yasuyuki; Desyatkin, Roman V; Hatano, Ryusuke; Tahara, Satoshi

    2009-05-01

    For evaluating N(2) fixation of diazotrophic bacteria, nitrogen-poor liquid media supplemented with at least 0.5% sugar and 0.2% agar are widely used for acetylene reduction assays. In such a soft gel medium, however, many N(2)-fixing soil bacteria generally show only trace acetylene reduction activity. Here, we report that use of a N(2) fixation medium solidified with gellan gum instead of agar promoted growth of some gellan-preferring soil bacteria. In a soft gel medium solidified with 0.3% gellan gum under appropriate culture conditions, bacterial microbiota from boreal forest bed soils and some free-living N(2)-fixing soil bacteria isolated from the microbiota exhibited 10- to 200-fold-higher acetylene reduction than those cultured in 0.2% agar medium. To determine the N(2) fixation-activating mechanism of gellan gum medium, qualitative differences in the colony-forming bacterial components from tested soil microbiota were investigated in plate cultures solidified with either agar or gellan gum for use with modified Winogradsky's medium. On 1.5% agar plates, apparently cryophilic bacterial microbiota showed strictly distinguishable microbiota according to the depth of soil in samples from an eastern Siberian Taiga forest bed. Some pure cultures of proteobacteria, such as Pseudomonas fluorescens and Burkholderia xenovorans, showed remarkable acetylene reduction. On plates solidified with 1.0% gellan gum, some soil bacteria, including Luteibacter sp., Janthinobacterium sp., Paenibacillus sp., and Arthrobacter sp., uniquely grew that had not grown in the presence of the same inoculants on agar plates. In contrast, Pseudomonas spp. and Burkholderia spp. were apparent only as minor colonies on the gellan gum plates. Moreover, only gellan gum plates allowed some bacteria, particularly those isolated from the shallow organic soil layer, to actively swarm. In consequence, gellan gum is a useful gel matrix to bring out growth potential capabilities of many soil

  2. 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...

  3. Antimicrobial assays of natural extracts and their inhibitory effect against Listeria innocua and fish spoilage bacteria, after incorporation into biopolymer edible films.

    Science.gov (United States)

    Iturriaga, L; Olabarrieta, I; de Marañón, I Martínez

    2012-08-01

    The antimicrobial activity of twelve natural extracts was tested against two fish spoilage bacteria (Pseudomonas fluorescens and Aeromonas hydrophila/caviae) and Listeria innocua, in order to assess their potential utilization in the preservation and safety of minimally processed fish products. After a screening of the active extracts by agar diffusion and vapour diffusion methods, oregano and thyme essential oils and citrus extract were selected. The minimum inhibitory concentration (MIC) of the selected extracts was determined by disc diffusion method against target bacteria and at two temperatures: bacteria's optimal growth temperature (30 °C or 37 °C) and refrigeration temperature (4 °C). Due to its better solubility, lack of odour and greater inhibitory effect obtained against L. innocua at refrigerated temperature, citrus extract was selected and incorporated at 1% (v/v) into different biopolymer film forming solutions (gelatin, methyl cellulose and their blend 50:50 w/w). The antimicrobial activity of the developed films was then evaluated, just after preparation of the films and after one month of storage at 43±3% relative humidity and 24±3 °C. Regardless of the biopolymer matrix, all the developed films showed antimicrobial activity against the target bacteria. The most sensitive bacterium towards active films was L. innocua while P. fluorescens appeared as the most resistant one, in accordance with the previously performed antimicrobial tests for pure extracts. The differences in activity of the films between the tested two temperatures were not significant except for L. innocua, for which three times higher inhibition diameters were observed at refrigerated temperature. The inhibitory effectiveness of the films against the tested strains was maintained regardless of the biopolymer matrix for at least one month. Therefore, these edible films show potential for their future use in fresh fish fillets preservation. Copyright © 2012 Elsevier B.V. All

  4. Isolation of Bacteria with Antifungal Activity against the Phytopathogenic Fungi Stenocarpella maydis and Stenocarpella macrospora

    Science.gov (United States)

    Petatán-Sagahón, Iván; Anducho-Reyes, Miguel Angel; Silva-Rojas, Hilda Victoria; Arana-Cuenca, Ainhoa; Tellez-Jurado, Alejandro; Cárdenas-Álvarez, Isabel Oyuki; Mercado-Flores, Yuridia

    2011-01-01

    Stenocarpella maydis and Stenocarpella macrospora are the causal agents of ear rot in corn, which is one of the most destructive diseases in this crop worldwide. These fungi are important mycotoxin producers that cause different pathologies in farmed animals and represent an important risk for humans. In this work, 160 strains were isolated from soil of corn crops of which 10 showed antifungal activity against these phytopathogens, which, were identified as: Bacillus subtilis, Pseudomonas spp., Pseudomonas fluorescens, and Pantoea agglomerans by sequencing of 16S rRNA gene and the phylogenetic analysis. From cultures of each strain, extracellular filtrates were obtained and assayed to determine antifungal activity. The best filtrates were obtained in the stationary phase of B. subtilis cultures that were stable to the temperature and extreme pH values; in addition they did not show a cytotoxicity effect against brine shrimp and inhibited germination of conidia. The bacteria described in this work have the potential to be used in the control of white ear rot disease. PMID:22016606

  5. Generation and initial characterization of Pseudomonas stutzeri KC mutants with impaired ability to degrade carbon tetrachloride.

    Science.gov (United States)

    Sepúlveda-Torres, L C; Rajendran, N; Dybas, M J; Criddle, C S

    1999-01-01

    Under iron-limiting conditions, Pseudomonas stutzeri KC secretes a small but as yet unidentified factor that transforms carbon tetrachloride (CT) to CO2 and nonvolatile products when activated by reduction at cell membranes. Pseudomonas fluorescens and other cell types activate the factor. Triparental mating was used to generate kanamycin-resistant lux::Tn5 recombinants of strain KC. Recombinants were streaked onto the surface of agar medium plugs in microtiter plates and were then screened for carbon tetrachloride degradation by exposing the plates to gaseous 14C-carbon tetrachloride. CT+ recombinants generated nonvolatile 14C-labeled products, but four CT- recombinants did not generate significant nonvolatile 14C-labeled products and had lost the ability to degrade carbon tetrachloride. When colonies of P. fluorescens were grown next to colonies of CT+ recombinants and were exposed to gaseous 14C-carbon tetrachloride, 14C-labeled products accumulated around the P. fluorescens colonies, indicating that the factor secreted by CT+ colonies had diffused through the agar and become activated. When P. fluorescens was grown next to CT- colonies, little carbon tetrachloride transformation was observed, indicating a lack of active factor. Expression of lux reporter genes in three of the CT- mutants was regulated by added iron and was induced under the same iron-limiting conditions that induce carbon tetrachloride transformation in the wild-type.

  6. 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...

  7. 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...

  8. Non-susceptibility trends among Pseudomonas aeruginosa and other non-fermentative Gram-negative bacteria from bacteraemias in the UK and Ireland, 2001-06.

    Science.gov (United States)

    Livermore, David M; Hope, Russell; Brick, Geraldine; Lillie, Mark; Reynolds, Rosy

    2008-11-01

    Pseudomonas and Acinetobacter spp. are important opportunists, notorious for resistance. Pseudomonas spp. are collected in the British Society for Antimicrobial Chemotherapy (BSAC) bacteraemia surveillance, with Acinetobacter spp. and Stenotrophomonas maltophilia well represented in the 'other Gram-negatives' group. Data for collected isolates were reviewed together with LabBase bacteraemia reports to the Health Protection Agency (HPA). Isolates with unusual resistances were subjected to molecular investigation. From 2001 to 2006, the BSAC surveillance collected 1226 Pseudomonas aeruginosa, 240 Acinetobacter spp.-125 of them Acinetobacter calcoaceticus/baumannii (Acb) complex-and 165 S. maltophilia. Among P. aeruginosa, non-susceptibility rates to beta-lactams and gentamicin fluctuated, without trend, below 10%; those to ciprofloxacin ranged from 16% to 22%. One P. aeruginosa isolate from 2001 had VIM-2 metallo-beta-lactamase. For Acb, the BSAC data indicated frequent non-susceptibility, except to imipenem, where only five non-susceptible isolates were collected, all after 2003, four of them belonging to the OXA-23 clone 1 lineage which is prevalent in Southeast England. Reports to the HPA indicated rising imipenem non-susceptibility in Acb (P /=16 mg/L for Acb OXA-23 clone 1. Ceftobiprole had higher MICs than ceftazidime for P. aeruginosa, but 81% of the isolates were inhibited at

  9. Population dynamics of the fast-growing sub-populations of Pseudomonas and total bacteria, and their protozoan grazers, revealed by fenpropimorph treatment

    DEFF Research Database (Denmark)

    Thirup, L; Ekelund, Flemming; Johnsen, Kaare Eske

    2000-01-01

    The population dynamics of indigenous soil bacteria and protozoa on decaying barley roots were followed by using litter bags buried in laboratory-incubated soil. The soil was either non-treated or treated with the fungicide fenpropimorph (in the formulation Corbel) at concentrations corresponding...... of protozoa corresponding to the two subpopulations was followed. The results strongly indicate a predatory association between the protozoa and bacteria. This was shown by a tight temporal association, and by a stimulation of bacteria following predatory release when protozoa were inhibited by fenpropimorph...

  10. Physiological studies of methane- and methanol-oxidizing bacteria: comparison of a primary alcohol dehydrogenase from Methylococcus capsulatus (Texas strain) and Pseudomonas species M27.

    Science.gov (United States)

    Patel, R N; Bose, H R; Mandy, W J; Hoare, D S

    1972-05-01

    A primary alcohol dehydrogenase has been purified from Methylococcus capsulatus (Texas strain). The purified enzyme catalyzes the oxidation of methanol and formaldehyde to formate; other primary alcohols are oxidized to their corresponding aldehydes. Ammonium ions are required for enzyme activity. The enzyme has a molecular weight of 120,000 daltons and consists of two 62,000 molecular-weight subunits which dissociate at acidic pH. The enzyme is similar to an alcohol dehydrogenase enzyme isolated from Pseudomonas sp. M27.

  11. Interactions between biosurfactant-producing Pseudomonas and Phytophthora species

    NARCIS (Netherlands)

    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

  12. Clearance of Pseudomonas aeruginosa Foreign-Body Biofilm Infections through Reduction of the Cyclic Di-GMP Level in the Bacteria

    DEFF Research Database (Denmark)

    Christensen, Louise D.; van Gennip, Maria; Rybtke, Morten Theil

    2013-01-01

    Opportunistic pathogenic bacteria can engage in biofilm-based infections that evade immune responses and develop into chronic conditions. Because conventional antimicrobials cannot efficiently eradicate biofilms, there is an urgent need to develop alternative measures to combat biofilm infections...

  13. Translocation of bacteria from animal excrements to soil and associated habitats

    NARCIS (Netherlands)

    Kupriyanov, A.A.; Kunenkova, N.N.; Bruggen, van A.H.C.; Semenov, A.M.

    2009-01-01

    The population dynamics of Salmonella enterica var. Typhimurium MAE 110 gfp, Escherichia coli O157:H7 gfp, and Pseudomonas fluorescens 32 gfp were investigated in their introduction to cattle excrements and subsequent entering the soil, plants of cress (Lepidium sativum L.), and migration through

  14. Levels of Water-Soluble Vitamins in Methanogenic and Non-Methanogenic Bacteria

    OpenAIRE

    Leigh, John A.

    1983-01-01

    The levels of seven water-soluble vitamins in Methanobacterium thermoautotrophicum, Methanococcus voltae, Escherichia coli, Bacillus subtilis, Pseudomonas fluorescens, and Bacteroides thetaiotaomicron were compared by using a vitamin-requiring Leuconostoc strain. Both methanogens contained levels of folic acid and pantothenic acid which were approximately two orders of magnitude lower than levels in the nonmethanogens. Methanobacterium thermoautotrophicum contained levels of thiamine, biotin,...

  15. Development of dry gram-negative bacteria biocontrol products and small pilot tests against dry rot

    Science.gov (United States)

    Pseudomonas fluorescens strains S11:P:12, P22:Y:05, and S22:T:04 suppress four important storage potato maladies; dry rot, late blight, pink rot, and sprouting. Studies were designed to identify methods for producing a dried, efficacious biological control product. The strains were evaluated individ...

  16. 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.

  17. Walleye Autochthonous Bacteria as Promising Probiotic Candidates against Flavobacterium columnare

    Directory of Open Access Journals (Sweden)

    Hamza Seghouani

    2017-07-01

    Full Text Available Walleye (Sander vitreus is the second most fished freshwater species in Canada. While much sought by anglers, walleye also supports substantial commercial fisheries. To cope with the recent decline of wild walleye populations, fish farmers produce juveniles for lake stocking. However, walleye breeding is particularly tedious, mostly due to high disease susceptibility at larval and juvenile developmental stages. The main threat is the columnaris disease, which is caused by Flavobacterium columnare, an opportunistic bacteria. As F. columnare strains exhibit increasing antibiotic resistance, there is a strong need to develop efficient and sustainable alternative strategies to control columnaris disease. Bacterial probiotics have been shown to mitigate infections either by enhancing host immune response or by inhibiting pathogen growth. Being successfully assessed in many fish/pathogen combinations, we developed a tailored probiotic strategy for walleye to prevent and treat columnaris disease. Thirty-seven endogenous bacterial strains were isolated from healthy walleye’s skin and gut, were tested in vitro against F. columnare. Significant antagonistic effect against F. columnare was measured for 2 out of 37 endogenous strains. These two probiotic strains were identified as Pseudomonas fluorescens. The antagonistic effect of these two successful probiotics was further validated in vivo during a 2-month stress trial: groups receiving probiotic treatments showed on average 53.74% survival improvement.

  18. Pseudomonas aeruginosa Biofilms

    DEFF Research Database (Denmark)

    Alhede, Maria; Bjarnsholt, Thomas; Givskov, Michael

    2014-01-01

    The opportunistic gram-negative bacterium Pseudomonas aeruginosa is implicated in many chronic infections and is readily isolated from chronic wounds, medical devices, and the lungs of cystic fibrosis patients. P. aeruginosa is believed to persist in the host organism due to its capacity to form...... biofilms, which protect the aggregated, biopolymer-embedded bacteria from the detrimental actions of antibiotic treatments and host immunity. A key component in the protection against innate immunity is rhamnolipid, which is a quorum sensing (QS)-regulated virulence factor. QS is a cell-to-cell signaling...

  19. Inhibition of pathogenic and spoilage bacteria by a novel biofilm-forming Lactobacillus isolate: a potential host for the expression of heterologous proteins.

    Science.gov (United States)

    Jalilsood, Tannaz; Baradaran, Ali; Song, Adelene Ai-Lian; Foo, Hooi Ling; Mustafa, Shuhaimi; Saad, Wan Zuhainis; Yusoff, Khatijah; Rahim, Raha Abdul

    2015-07-07

    Bacterial biofilms are a preferred mode of growth for many types of microorganisms in their natural environments. The ability of pathogens to integrate within a biofilm is pivotal to their survival. The possibility of biofilm formation in Lactobacillus communities is also important in various industrial and medical settings. Lactobacilli can eliminate the colonization of different pathogenic microorganisms. Alternatively, new opportunities are now arising with the rapidly expanding potential of lactic acid bacteria biofilms as bio-control agents against food-borne pathogens. A new isolate Lactobacillus plantarum PA21 could form a strong biofilm in pure culture and in combination with several pathogenic and food-spoilage bacteria such as Salmonella enterica, Bacillus cereus, Pseudomonas fluorescens, and Aeromonas hydrophila. Exposure to Lb. plantarum PA21 significantly reduced the number of P. fluorescens, A. hydrophila and B. cereus cells in the biofilm over 2-, 4- and 6-day time periods. However, despite the reduction in S. enterica cells, this pathogen showed greater resistance in the presence of PA21 developed biofilm, either in the planktonic or biofilm phase. Lb. plantarum PA21 was also found to be able to constitutively express GFP when transformed with the expression vector pMG36e which harbors the gfp gene as a reporter demonstrating that the newly isolated strain can be used as host for genetic engineering. In this study, we evaluate the ability of a new Lactobacillus isolate to form strong biofilm, which would provide the inhibitory effect against several spoilage and pathogenic bacteria. This new isolate has the potential to serve as a safe and effective cell factory for recombinant proteins.

  20. Kinetics of Enantiomerically Enriched Synthesis of Solketal Esters Using Native and SBA-15 supported P. Fluorescens Lipase

    Directory of Open Access Journals (Sweden)

    Zniszczoł Aurelia

    2017-06-01

    Full Text Available The studies showed that alkaline lipase from Pseudomonas fluorescens enables an irreversible transesterification of vinyl esters to give enantiomeric excess (eeR of about 80% using vinyl butyrate as acyl donor and diisopropyl ether as a solvent, at partially optimized conditions. For the native lipase the process was adequately described by a five-parameter Ping-Pong Bi Bi model for both enantiomers plus expression accounting for the formation of enzyme-acyl donor complex, but for the same lipase supported on mesoporous materials of SBA-15-Oc type, R-product inhibition also had to be taken into account. The use of hydrophobic support increased by more than two-fold the rate of the S-solketal conversion but even more that of R-solketal. Thus the immobilization of lipase had very positive effect on the process kinetics but decreased its enantioselectivity.

  1. Specific interactions between arbuscular mycorrhizal fungi and plant growth-promoting bacteria: as revealed by different combinations.

    Science.gov (United States)

    Jäderlund, Lotta; Arthurson, Veronica; Granhall, Ulf; Jansson, Janet K

    2008-10-01

    The interactions between two plant growth-promoting rhizobacteria (PGPR, Pseudomonas fluorescens SBW25 and Paenibacillus brasilensis PB177), two arbuscular mycorrhizal (AM) fungi (Glomus mosseae and Glomus intraradices) and one pathogenic fungus (Microdochium nivale) were investigated on winter wheat (Triticum aestivum cultivar Tarso) in a greenhouse trial. PB177, but not SBW25, had strong inhibitory effects on M. nivale in dual culture plate assays. The results from the greenhouse experiment show very specific interactions; for example, the two AM fungi react differently when interacting with the same bacteria on plants. Glomus intraradices (single inoculation or together with SBW25) increased plant dry weight on M. nivale-infested plants, suggesting that the pathogenic fungus is counteracted by G. intraradices, but PB177 inhibited this positive effect. This is an example of two completely different reactions between the same AM fungus and two species of bacteria, previously known to enhance plant growth and inhibit pathogens. When searching for plant growth-promoting microorganisms, it is therefore important to test for the most suitable combination of plant, bacteria and fungi in order to achieve satisfactory plant growth benefits.

  2. Specific interactions between arbuscular mycorrhizal fungi and plant growth-promoting bacteria--as revealed by different combinations

    Energy Technology Data Exchange (ETDEWEB)

    Jaderlund, Lotta; Arthurson, Veronica; Granhall, Ulf; Jansson, Janet K.

    2008-05-15

    The interactions between two plant growth promoting rhizobacteria (PGPR), Pseudomonas fluorescens SBW25 and Paenibacillus brasilensis PB177, two arbuscular mycorrhizal (AM) fungi (Glomus mosseae and G. intraradices) and one pathogenic fungus (Microdochium nivale) were investigated on winter wheat (Triticum aestivum cultivar Tarso) in a greenhouse trial. PB177, but not SBW25, had strong inhibitory effects on M. nivale in dual culture plate assays. The results from the greenhouse experiment show very specific interactions; e.g. the two AM fungi react differently when interacting with the same bacteria on plants. G. intraradices (single inoculation or together with SBW25) increased plant dry weight on M. nivale infested plants, suggesting that the pathogenic fungus is counteracted by G. intraradices, but PB177 inhibited this positive effect. This is an example of two completely different reactions between the same AM fungus and two species of bacteria, previously known to enhance plant growth and inhibit pathogens. When searching for plant growth promoting microorganisms it is therefore important to test for the most suitable combination of plant, bacteria and fungi in order to get satisfactory plant growth benefits.

  3. Effect of electrical charges and fields on injury and viability of airborne bacteria.

    Science.gov (United States)

    Mainelis, Gediminas; Górny, Rafał L; Reponen, Tiina; Trunov, Mikhaylo; Grinshpun, Sergey A; Baron, Paul; Yadav, Jagjit; Willeke, Klaus

    2002-07-20

    In this study, the effects of the electric charges and fields on the viability of airborne microorganisms were investigated. The electric charges of different magnitude and polarity were imparted on airborne microbial cells by a means of induction charging. The airborne microorganisms carrying different electric charge levels were then extracted by an electric mobility analyzer and collected using a microbial sampler. It was found that the viability of Pseudomonas fluorescens bacteria, used as a model for sensitive bacteria, carrying a net charge from 4100 negative to 30 positive elementary charges ranged between 40% and 60%; the viability of the cells carrying >2700 positive charges was below 1.5%. In contrast, the viability of the stress-resistant spores of Bacillus subtilis var. niger (used as simulant of anthrax-causing Bacillus anthracis spores when testing bioaerosol sensors in various studies), was not affected by the amount of electric charges on the spores. Because bacterial cells depend on their membrane potential for basic metabolic activities, drastic changes occurring in the membrane potential during aerosolization and the local electric fields induced by the imposed charges appeared to affect the sensitive cells' viability. These findings facilitate applications of electric charging for environmental control purposes involving sterilization of bacterial cells by imposing high electric charges on them. The findings from this study can also be used in the development of new bioaerosol sampling methods based on electrostatic principles. Copyright 2002 Wiley Periodicals, Inc.

  4. Isolation and Identification of Sodium Fluoroacetate Degrading Bacteria from Caprine Rumen in Brazil

    Directory of Open Access Journals (Sweden)

    Expedito K. A. Camboim

    2012-01-01

    Full Text Available The objective of this paper was to report the isolation of two fluoroacetate degrading bacteria from the rumen of goats. The animals were adult goats, males, crossbred, with rumen fistula, fed with hay, and native pasture. The rumen fluid was obtained through the rumen fistula and immediately was inoculated 100 μL in mineral medium added with 20 mmol L−1 sodium fluoroacetate (SF, incubated at 39°C in an orbital shaker. Pseudomonas fluorescens (strain DSM 8341 was used as positive control for fluoroacetate dehalogenase activity. Two isolates were identified by 16S rRNA gene sequencing as Pigmentiphaga kullae (ECPB08 and Ancylobacter dichloromethanicus (ECPB09. These bacteria degraded sodium fluoroacetate, releasing 20 mmol L−1 of fluoride ion after 32 hours of incubation in Brunner medium containing 20 mmol L−1 of SF. There are no previous reports of fluoroacetate dehalogenase activity for P. kullae and A. dichloromethanicus. Control measures to prevent plant intoxication, including use of fences, herbicides, or other methods of eliminating poisonous plants, have been unsuccessful to avoid poisoning by fluoroacetate containing plants in Brazil. In this way, P. kullae and A. dichloromethanicus may be used to colonize the rumen of susceptible animals to avoid intoxication by fluoroacetate containing plants.

  5. Ribosomal RNA content in microcolony forming soil bacteria measured by quantitative 16S rRNA hybridization and image analysis

    NARCIS (Netherlands)

    Binnerup, S.J.; Bloem, J.; Hansen, B.M.; Wolters, W.; Veninga, M.; Hansen, M.

    2001-01-01

    The correlation between the growth rate, rRNA concentration and number of rrna operons was studied in Alcaligenes sp. A2, Pseudomonas fluorescens R2f and Bacillus sp. B1 cells grown exponentially in liquid 1/10 strength tryptic soy broth (1/10 TSB) medium and grown to microcolony (mCFU) size on

  6. Formation of dry gram-negative bacteria biocontrol products and small pilot tests against potato dry rot

    Science.gov (United States)

    Pseudomonas fluorescens strains S11:P:12, P22:Y:05, and S22:T:04 reduce important potato maladies in storage including dry rot, late blight, pink rot, and sprouting. Experiments were conducted to identify methods for producing a dried, efficacious biological control product from one or more of these...

  7. Effect of Catechins, Green tea Extract and Methylxanthines in Combination with Gentamicin Against Staphylococcus aureus and Pseudomonas aeruginosa - Combination therapy against resistant bacteria -

    Directory of Open Access Journals (Sweden)

    Bibi Sedigheh Fazly Bazzaz

    2016-12-01

    Full Text Available Objectives: Bacterial resistant infections have become a global health challenge and threaten the society’s health. Thus, an urgent need exists to find ways to combat resistant pathogens. One promising approach to overcoming bacterial resistance is the use of herbal products. Green tea catechins, the major green tea polyphenols, show antimicrobial activity against resistant pathogens. The present study aimed to investigate the effect of catechins, green tea extract, and methylxanthines in combination with gentamicin against standard and clinical isolates of Staphylococcus aureus (S. aureus and the standard strain of Pseudomonas aeruginosa (P. aeruginosa. Methods: The minimum inhibitory concentration (MIC and the minimum bactericidal concentration (MBC values of different agents against bacterial strains were determined. The interactions of green tea extract, epigallate catechin, epigallocatechin gallate, two types of methylxanthine, caffeine, and theophylline with gentamicin were studied in vitro by using a checkerboard method and calculating the fraction inhibitory concentration index (FICI. Results: The MICs of gentamicin against bacterial strains were in the range of 0.312 - 320 μg/mL. The MIC values of both types of catechins were 62.5 - 250 μg/ mL. Green tea extract showed insufficient antibacterial activity when used alone. Methylxanthines had no intrinsic inhibitory activity against any of the bacterial strains tested. When green tea extract and catechins were combined with gentamicin, the MIC values of gentamicin against the standard strains and a clinical isolate were reduced, and synergistic activities were observed (FICI < 1. A combination of caffeine with gentamicin did not alter the MIC values of gentamicin. Conclusion: The results of the present study revealed that green tea extract and catechins potentiated the antimicrobial action of gentamicin against some clinical isolates of S. aureus and standard P. aeruginosa strains

  8. Multi-functional characteristics of the Pseudomonas aeruginosa type III needle-tip protein, PcrV; comparison to orthologs in other gram negative bacteria

    Directory of Open Access Journals (Sweden)

    Hiromi eSato

    2011-07-01

    Full Text Available Pseudomonas aeruginosa possesses a type III secretion system (T3SS to intoxicate host cells and evade innate immunity. This virulence-related machinery consists of a molecular syringe and needle assembled on the bacterial surface, which allows delivery of T3 effector proteins into infected cells. To accomplish a one-step effector translocation, a tip protein is required at the top end of the T3 needle structure. Strains lacking expression of the functional tip protein fail to intoxicate host cells.P. aeruginosa encodes a T3S that is highly homologous to the proteins encoded by Yersinia species. The needle tip proteins of Yersinia, LcrV, and P. aeruginosa, PcrV, share 37% identity and 65% similarity. Other known tip proteins are AcrV (Aeromonas, IpaD (Shigella, SipD (Salmonella, BipD (Burkholderia, EspA (EPEC, EHEC, Bsp22 (Bordetella, with additional proteins identified from various Gram negative species, such as Vibrio and Bordetella. The tip proteins can serve as a protective antigen or may be critical for sensing host cells and evading innate immune responses. Recognition of the host microenvironment transcriptionally activates synthesis of T3SS components. The machinery appears to be mechanically controlled by the assemblage of specific junctions within the apparatus. These junctions include the tip and base of the T3 apparatus, the needle proteins and components within the bacterial cytoplasm. The tip proteins likely have chaperone functions for translocon proteins, allowing the proper assembly of translocation channels in the host membrane and completing vectorial delivery of effector proteins into the host cytoplasm. Multifunctional features of the needle-tip proteins appear to be intricately controlled. In this review, we highlight the functional aspects and complex controls of T3 needle-tip proteins with particular emphasis on PcrV and LcrV.

  9. Transcriptomic profiling of microbe-microbe interactions reveals the specific response of the biocontrol strain P. fluorescens In5 to the phytopathogen Rhizoctonia solani

    DEFF Research Database (Denmark)

    Hennessy, Rosanna Catherine; Glaring, Mikkel Andreas; Olsson, Stefan

    2017-01-01

    metabolites, e.g. non-ribosomal synthetases and hydrogen cyanide were not differentially expressed at the time points studied. CONCLUSION: This study demonstrates that genes possibly involved in metabolite detoxification are highly upregulated in P. fluorescens In5 when co-cultured with plant pathogens...... and in particular the fungus R. solani. This highlights the importance of studying microbe-microbe interactions to gain a better understanding of how different systems function in vitro and ultimately in natural systems where biocontrol agents can be used for the sustainable management of plant diseases.......BACKGROUND: Few studies to date report the transcriptional response of biocontrol bacteria toward phytopathogens. In order to gain insights into the potential mechanism underlying the antagonism of the antimicrobial producing strain P. fluorescens In5 against the phytopathogens Rhizoctonia solani...

  10. Display of a thermostable lipase on the surface of a solvent-resistant bacterium, Pseudomonas putida GM730, and its applications in whole-cell biocatalysis

    Directory of Open Access Journals (Sweden)

    Kwon Seok-Joon

    2006-04-01

    Full Text Available Abstract Background Whole-cell biocatalysis in organic solvents has been widely applied to industrial bioprocesses. In two-phase water-solvent processes, substrate conversion yields and volumetric productivities can be limited by the toxicity of solvents to host cells and by the low mass transfer rates of the substrates from the solvent phase to the whole-cell biocatalysts in water. Results To solve the problem of solvent toxicity, we immobilized a thermostable lipase (TliA from Pseudomonas fluorescens on the cell surface of a solvent-resistant bacterium, Pseudomonas putida GM730. Surface immobilization of enzymes eliminates the mass-transfer limitation imposed by the cell wall and membranes. TliA was successfully immobilized on the surface of P. putida cells using the ice-nucleation protein (INP anchoring motif from Pseudomonas syrinage. The surface location was confirmed by flow cytometry, protease accessibility and whole-cell enzyme activity using a membrane-impermeable substrate. Three hundred and fifty units of whole-cell hydrolytic activity per gram dry cell mass were obtained when the enzyme was immobilized with a shorter INP anchoring motif (INPNC. The surface-immobilized TliA retained full enzyme activity in a two-phase water-isooctane reaction system after incubation at 37°C for 12 h, while the activity of the free form enzyme decreased to 65% of its initial value. Whole cells presenting immobilized TliA were shown to catalyze three representative lipase reactions: hydrolysis of olive oil, synthesis of triacylglycerol and chiral resolution. Conclusion In vivo surface immobilization of enzymes on solvent-resistant bacteria was demonstrated, and appears to be useful for a variety of whole-cell bioconversions in the presence of organic solvents.

  11. The Mechanism of Killing by the Proline-Rich Peptide Bac7(1-35) against Clinical Strains of Pseudomonas aeruginosa Differs from That against Other Gram-Negative Bacteria.

    Science.gov (United States)

    Runti, Giulia; Benincasa, Monica; Giuffrida, Grazia; Devescovi, Giulia; Venturi, Vittorio; Gennaro, Renato; Scocchi, Marco

    2017-04-01

    Pseudomonas aeruginosa infections represent a serious threat to worldwide health. Proline-rich antimicrobial peptides (PR-AMPs), a particular group of peptide antibiotics, have demonstrated in vitro activity against P. aeruginosa strains. Here we show that the mammalian PR-AMP Bac7(1-35) is active against some multidrug-resistant cystic fibrosis isolates of P. aeruginosa By confocal microscopy and cytometric analyses, we investigated the mechanism of killing against P. aeruginosa strain PAO1 and three selected isolates, and we observed that the peptide inactivated the target cells by disrupting their cellular membranes. This effect is deeply different from that previously described for PR-AMPs in Escherichia coli and Salmonella enterica serovar Typhimurium, where these peptides act intracellularly after having been internalized by means of the transporter SbmA without membranolytic effects. The heterologous expression of SbmA in PAO1 cells enhanced the internalization of Bac7(1-35) into the cytoplasm, making the bacteria more susceptible to the peptide but at the same time more resistant to the membrane lysis, similarly to what occurs in E. coli The results evidenced a new mechanism of action for PR-AMPs and indicate that Bac7 has multiple and variable modes of action that depend on the characteristics of the different target species and the possibility to be internalized by bacterial transporters. This feature broadens the spectrum of activity of the peptide and makes the development of peptide-resistant bacteria a more difficult process. Copyright © 2017 American Society for Microbiology.

  12. 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.

  13. Variation in the Presence of Anti-Batrachochytrium dendrobatidis Bacteria of Amphibians Across Life Stages and Elevations in Ecuador.

    Science.gov (United States)

    Bresciano, J C; Salvador, C A; Paz-Y-Miño, C; Parody-Merino, A M; Bosch, J; Woodhams, D C

    2015-06-01

    Amphibian populations are decreasing worldwide due to a variety of factors. In South America, the chytrid fungus Batrachochytrium dendrobatidis (Bd) is linked to many population declines. The pathogenic effect of Bd on amphibians can be inhibited by specific bacteria present on host skin. This symbiotic association allows some amphibians to resist the development of the disease chytridiomycosis. Here, we aimed (1) to determine for the first time if specific anti-Bd bacteria are present on amphibians in the Andes of Ecuador, (2) to monitor anti-Bd bacteria across developmental stages in a focal amphibian, the Andean marsupial tree frog, Gastrotheca riobambae, that deposits larvae in aquatic habitats, and (3) to compare the Bd presence associated with host assemblages including 10 species at sites ranging in biogeography from Amazonian rainforest (450 masl) to Andes montane rainforest (3200 masl). We sampled and identified skin-associated bacteria of frogs in the field using swabs and a novel methodology of aerobic counting plates, and a combination of morphological, biochemical, and molecular identification techniques. The following anti-Bd bacteria were identified and found to be shared among several hosts at high-elevation sites where Bd was present at a prevalence of 32.5%: Janthinobacterium lividum, Pseudomonas fluorescens, and Serratia sp. Bd were detected in Gastrotheca spp. and not detected in the lowlands (sites below 1000 masl). In G. riobambae, recognized Bd-resistant bacteria start to be present at the metamorphic stage. Overall bacterial abundance was significantly higher post-metamorphosis and on species sampled at lower elevations. Further metagenomic studies are needed to evaluate the roles of host identity, life-history stage, and biogeography of the microbiota and their function in disease resistance.

  14. Effects of atmospheric conditions on ice nucleation activity of Pseudomonas

    Directory of Open Access Journals (Sweden)

    C. Glaux

    2012-11-01

    Full Text Available Although ice nuclei from bacterial origin are known to be efficient at the highest temperatures known for ice catalysts, quantitative data are still needed to assess their role in cloud processes. Here we studied the effects of three typical cloud conditions (i acidic pH (ii NO2 and O3 exposure and (iii UV-A exposure on the ice nucleation activity (INA of four Pseudomonas strains. Three of the Pseudomonas syringae strains were isolated from cloud water and the phyllosphere and Pseudomonas fluorescens strain CGina-01 was isolated from Antarctic glacier ice melt. Among the three conditions tested, acidic pH caused the most significant effects on INA likely due to denaturation of the ice nucleation protein complex. Exposure to NO2 and O3 gases had no significant or only weak effects on the INA of two P. syringae strains whereas the INA of P. fluorescens CGina-01 was significantly affected. The INA of the third P. syringae strain showed variable responses to NO2 and O3 exposure. These differences in the INA of different Pseudomonas suggest that the response to atmospheric conditions could be strain-specific. After UV-A exposure, a substantial loss of viability of all four strains was observed whereas their INA decreased only slightly. This corroborates the notion that under certain conditions dead bacterial cells can maintain their INA. Overall, the negative effects of the three environmental factors on INA were more significant at the warmer temperatures. Our results suggest that in clouds where temperatures are near 0 °C, the importance of bacterial ice nucleation in precipitation processes could be reduced by some environmental factors.

  15. 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

  16. Characterization of heavy metal-resistant endophytic bacteria from rape (Brassica napus) roots and their potential in promoting the growth and lead accumulation of rape

    Energy Technology Data Exchange (ETDEWEB)

    Sheng Xiafang [MOA Key Laboratory of Microbiological Engineering of Agricultural Environment, College of Life Science, Nanjing Agricultural University, Nanjing 210095 (China)], E-mail: xfsheng604@sohu.com; Xia Juanjuan; Jiang Chunyu; He Linyan; Qian Meng [MOA Key Laboratory of Microbiological Engineering of Agricultural Environment, College of Life Science, Nanjing Agricultural University, Nanjing 210095 (China)

    2008-12-15

    Two lead (Pb)-resistant endophytic bacteria were isolated from rape roots grown in heavy metal-contaminated soils and characterized. A pot experiment was conducted for investigating the capability of the two isolates to promote the growth and Pb uptake of rape from Pb-amended soil. The two isolates were identified as Pseudomonas fluorescens G10 and Microbacterium sp. G16 based on the 16S rDNA gene sequence analysis. Strains G10 and G16 exhibited different multiple heavy metal and antibiotic resistance characteristics and increased water-soluble Pb in solution and in Pb-added soil. Root elongation assays demonstrated increases in root elongation of inoculated rape seedlings compared to the control plants. Strain G16 produced indole acetic acid, siderophores and 1-aminocyclopropane-1-carboxylate deaminase. Increases in biomass production and total Pb uptake in the bacteria-inoculated plants were obtained compared to the control. The two strains could colonize the root interior and rhizosphere soil of rape after root inoculation. - Heavy metal-resistant endophytic bacteria from rape have the potential of promoting the growth and lead uptake of rape.

  17. 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.

  18. A study on inhibitory effects of Siğla tree (Liquidambar orientalis Mill. var. orientalis) storax against several bacteria.

    Science.gov (United States)

    Sağdiç, Osman; Ozkan, Gülcan; Ozcan, Musa; Ozçelik, Sami

    2005-06-01

    In this study, Siğla (Liquidambar orientalis Mill.) storax (styrax) was investigated for antibacterial activity against Aeromonas hydrophila, Bacillus amyloliquefaciens, B. brevis, B. cereus, B. megaterium, B. subtilis, Corynebacterium xerosis, Enterobacter aerogenes, Enterococcus faecalis, Escherichia coli, E. coli O157:H7, Klebsiella pneumoniae, Listeria monocytogenes, Micrococcus luteus, Mycobacterium smegmatis, Proteus vulgaris, Pseudomonas aeruginosa, P. fluorescens, Staphylococcus aureus and Yersinia enterocolitica. The storax was dissolved in absolute ethanol and was tested at concentrations of 10.0%, 1.0%, 0.4%, 0.2% and 0.1%. Pure ethanol was used for the control. The antibacterial activity of the Siğla storax was determined by using the agar diffusion method. The growths of A. hydrophila, B. amyloliquefaciens, B. megaterium, E. coli, E. coli O157:H7, L. monocytogenes and Y. enterocolitica were not inhibited by any of the concentrations of Siğla storax. The results showed that Siğla storax has antibacterial activity against many bacteria at concentrations of 10.0% and against some bacteria at concentrations of 1.0%, 0.4% and 0.2%. Copyright (c) 2005 John Wiley & Sons, Ltd.

  19. On the reaction of some bacteria and fungi on coal tar creosote. Zur Verhalten einiger Bakterien und Pilze gegenueber Steinkohlenteeroel

    Energy Technology Data Exchange (ETDEWEB)

    Schmidt, O.; Dittberner, D.; Faix, O. (Universitaet Hamburg, Hamburg (Germany). Ordinariat fuer Holzbiologie)

    1991-01-01

    To contribute to the waste management of wood preservatives, the biodegradability of coal tar creosote by bacteria and fungi has been investigated. Microorganisms comprised 24 bacterial strains and 31 fungi from different systematic and ecological groups as well as isolates from contaminated soils. Based on countings of viable cells, the experiments with various nutrient media, methods of cultivation, preservative concentrations, and organic solvents yielded some bacteria which could grow in the presence of creosote: {ital Aeromonas hydrophila}, {ital Flavobacterium} sp., {ital Pseudomonas arvilla}, {ital P. fluorescens}, and {ital P. putida}. The white-rot fungi {ital Bjerkandera adusta}, {ital Heterobasidion annosum}, {ital Hirschioporus abietinus}, {ital Lentinula edodes}, {ital Peniophora gigantea}, {ital Pleurotus ostreatus}, {ital Schizophyllum commune}, and {ital Trametes versicolor}, the brown-rot fungus {ital Lentinus lepideus}, the staining fungi {ital Ceratocystis piceae} and {ital Stereum sanguinolentum}, and the moulds {ital Paecilomyces variotii} and {ital Trichoderma viride} also grew with creosote. To prepare samples for IR-measurements, continuous extraction of creosote from the nutrient liquid by percolation with methylene chloride was suitable. However, the IR-spectra of creosote did not show any measurable changes after incubation with 16 bacterial strains and 6 fungi. 42 refs., 2 figs., 4 tabs.

  20. 76 FR 52871 - Pseudomonas fluorescens Strain CL145A; Exemption From the Requirement of a Tolerance

    Science.gov (United States)

    2011-08-24

    ... with plants, including those food plants consumed raw. The Manual of Clinical Microbiology (8th edition... a joint U.N. Food and Agriculture Organization/World Health Organization food standards program, and...

  1. Heterologous Expression of a Chitinase Gene From Aeromonas Caviaein Pseudomonas Fluorescens

    OpenAIRE

    Suwanto, Antonius; Malik, Amarila; TJAHJONO, BUDI; Harling, Rob

    2003-01-01

    A transcriptional fusion for an Aeromonas caviae chitinase gene was constructed under the control of a constitutive promoter of the kanaraycin resistance gene (PKmR). The construct was inserted into a medium copy number broad host range plasmid vector to yield recombinant plasmid pAM340, which harbored transcriptional fusion PKmR- chi. Another transcriptional fusion, Ptac-chi, in a recombinant plasmid pAM630, was conducted as comparison. Triparental mating of E. coli carrying the recombinant...

  2. HETEROLOGOUS EXPRESSION OF A CHITINASE GENE FROM AEROMONAS CAVIAEIN PSEUDOMONAS FLUORESCENS

    OpenAIRE

    ANTONIUS SUWANTO; AMARILA MAHK; BUDI TJAHJONO

    2003-01-01

    A transcriptional fusion for an Aeromonas caviae chitinase gene was constructed under the control of a constitutive promoter of the kanaraycin resistance gene (PKmR). The construct was inserted into a medium copy number broad host range plasmid vector to yield recombinant plasmid pAM340, which harbored transcriptional fusion PKmR- chi. Another transcriptional fusion, Ptac-chi, in a recombinant plasmid pAM630, was conducted as comparison. Triparental mating of E. coli carrying the recom...

  3. 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 st...

  4. New strategies for genetic engineering Pseudomonas syringae using recombination

    Science.gov (United States)

    Here we report that DNA oligonucleotides (oligos) introduced directly into bacteria by electroporation can recombine with the bacterial chromosome. This phenomenon was identified in Pseudomonas syringae and we subsequently found that Escherichia coli, Salmonella typhimurium and Shigella flexneri are...

  5. Biodegradation of endosulfan by mixed bacteria culture strains of ...

    African Journals Online (AJOL)

    Biodegradation of endosulfan by mixed bacteria culture strains of Pseudomonas aeruginosa and Staphylococcus aureus. Nsidibeabasi Calvin Nwokem, Calvin Onyedika Nwokem, Casmir Emmanuel Gimba, Beatrice Nkiruka Iwuala ...

  6. Bioalteration of synthetic Fe(III)-, Fe(II)-bearing basaltic glasses and Fe-free glass in the presence of the heterotrophic bacteria strain Pseudomonas aeruginosa: Impact of siderophores

    Science.gov (United States)

    Perez, Anne; Rossano, Stéphanie; Trcera, Nicolas; Huguenot, David; Fourdrin, Chloé; Verney-Carron, Aurélie; van Hullebusch, Eric D.; Guyot, François

    2016-09-01

    This study aims to evaluate the role of micro-organisms and their siderophores in the first steps of the alteration processes of basaltic glasses in aqueous media. In this regard, three different types of glasses - with or without iron, in the reduced Fe(II) or oxidized Fe(III) states - were prepared on the basis of a simplified basaltic glass composition. Control and Pseudomonas aeruginosa inoculated experiments were performed in a buffered (pH 6.5) nutrient depleted medium to stimulate the production of the pyoverdine siderophore. Results show that the presence of P. aeruginosa has an effect on the dissolution kinetics of all glasses as most of the calculated elemental release rates are increased compared to sterile conditions. Reciprocally, the composition of the glass in contact with P. aeruginosa has an impact on the bacterial growth and siderophore production. As an essential nutrient for this microbial strain, Fe notably appears to play a central role during biotic experiments. Its presence in the glass stimulates the bacterial growth and minimizes the synthesis of pyoverdine. Moreover the initial Fe2+/Fe3+ ratio in the glasses modulates this synthesis, as pyoverdine is not detected at all in the system in contact with Fe(III)-bearing glass. Finally, the dissolution rates appear to be correlated to siderophore concentrations as they increase with respect to sterile experiments in the order Fe(III)-bearing glass siderophores and Fe or Al for Fe(II)-bearing glass or Fe-free glass, respectively. The dissolution of an Fe-free glass is significantly improved in the presence of bacteria, as initial dissolution rates are increased by a factor of 3. This study attests to the essential role of siderophores in the P. aeruginosa-promoted dissolution processes of basaltic glasses as well as to the complex relationships between the nutritional potential of the glass and its dissolution rates.

  7. Pseudomonas spp. ISOLATED FROM THE ORAL CAVITY OF HEALTHCARE WORKERS FROM AN ONCOLOGY HOSPITAL IN MIDWESTERN BRAZIL

    Science.gov (United States)

    LIMA, Ana Beatriz Mori; LEÃO-VASCONCELOS, Lara Stefânia Netto de Oliveira; COSTA, Dayane de Melo; VILEFORT, Larissa Oliveira Rocha; ANDRÉ, Maria Cláudia Dantas Porfírio Borges; BARBOSA, Maria Alves; PRADO-PALOS, Marinésia Aparecida

    2015-01-01

    This cross-sectional study, performed in an oncology hospital in Goiania, aimed to characterize the prevalence of oral colonization and antimicrobial susceptibility of Pseudomonas spp. isolated from the saliva of healthcare workers. Microorganisms were subjected to biochemical tests, susceptibility profile, and phenotypic detection. Of 76 participants colonized with Gram negative bacilli, 12 (15.8%) harbored Pseudomonas spp. Of all isolates, P. aeruginosa (75.0%), P. stutzeri (16.7%), and P. fluorescens (8.3%), were resistant to cefoxitin, and therefore likely to be AmpC producers. The results are clinically relevant and emphasize the importance of surveillance to minimize bacterial dissemination and multiresistance. PMID:27049706

  8. Multiple antibiotic-resistant bacteria on fluted pumpkin leaves, a herb of therapeutic value.

    Science.gov (United States)

    Igbeneghu, Oluwatoyin A; Abdu, Abdulrasheed B

    2014-06-01

    Fluted pumpkin (Telfairia occidentalis) is a minimally-processed green leafy vegetable traditionally used for its antianaemic properties in the form of leaf juice without a heating or inactivation step before consumption. The aim of the study was to assess the presence of surface microbiota on T. occidentalis leaves and also to determine the antimicrobial susceptibility of isolated organisms. Bacterial contaminants on 50 samples of T. occidentalis leaves were isolated and characterized using standard biochemical methods and the antimicrobial susceptibility of isolated organisms was determined using the antibiotic disc diffusion assay. The results obtained show that the leaves of T. occidentalis is contaminated with organisms which included Enterobacter agglomerans (25.9%), Proteus vulgaris (24.9%), Klebsiella spp. (2.6%), and Serratia liquefaciens (2.1%). Other bacterial isolates recovered in order of frequency included: Staphylococcus spp. (33.7%), Bacillus spp. (8.3%), and Pseudomonas fluorescens (2.6%). Of the 193 bacterial isolates from the leaves of T. occidentalis samples tested for antimicrobial resistance, all (100%) were found to be resistant to ampicillin, cloxacillin, augmentin, erythromycin, and tetracycline while 96% of the isolates were resistant to cephalothin. Resistance to trimethoprim (93%) and gentamicin (83%) was also observed. Approximately, 22% of the isolates were resistant to ciprofloxacin; however, only 11 (5.8%) were resistant to ofloxacin. Thus, uncooked T. occidentalis is a potential source of highly-resistant epiphytic bacteria which could be opportunistic pathogens in consumers.

  9. Interaction of selected actinides (U, Cm) with bacteria relevant to nuclear waste disposal

    Energy Technology Data Exchange (ETDEWEB)

    Luetke, Laura

    2013-07-01

    To assess the safety of a site destined for storage of nuclear waste enhanced research effort is demanded to investigate the complex interactions of released radionuclides with parts of the environment that includes indigenous microorganisms. This work aimed at assessing the interactions of two bacterial strains with the actinides uranium and curium with a focus on thermodynamics to provide stability constants of the actinide bacteria species formed usable for modelling the distribution of these actinides in the environment. The influences of Pseudomonas fluorescens (CCUG 32456A) isolated from the granitic aquifers at Aespoe(Sweden) and a novel isolate from Mont Terri Opalinus clay (Switzerland), Paenibacillus sp. MT-2.2, were investigated. A combined approach using microbiological and spectroscopic techniques as well as potentiometry was employed to characterize the U(VI) and Cm(III) binding onto the cell surface functional groups structurally and thermodynamically. Further, due to its similar ionic radius to Cm(III) also Eu(III) was studied as non-radioactive analog.

  10. Proposal for a method to estimate nutrient shock effects in bacteria

    Directory of Open Access Journals (Sweden)

    Azevedo Nuno F

    2012-08-01

    Full Text Available Abstract Background Plating methods are still the golden standard in microbiology; however, some studies have shown that these techniques can underestimate the microbial concentrations and diversity. A nutrient shock is one of the mechanisms proposed to explain this phenomenon. In this study, a tentative method to assess nutrient shock effects was tested. Findings To estimate the extent of nutrient shock effects, two strains isolated from tap water (Sphingomonas capsulata and Methylobacterium sp. and two culture collection strains (E. coli CECT 434 and Pseudomonas fluorescens ATCC 13525 were exposed both to low and high nutrient conditions for different times and then placed in low nutrient medium (R2A and rich nutrient medium (TSA. The average improvement (A.I. of recovery between R2A and TSA for the different times was calculated to more simply assess the difference obtained in culturability between each medium. As expected, A.I. was higher when cells were plated after the exposition to water than when they were recovered from high-nutrient medium showing the existence of a nutrient shock for the diverse bacteria used. S. capsulata was the species most affected by this phenomenon. Conclusions This work provides a method to consistently determine the extent of nutrient shock effects on different microorganisms and hence quantify the ability of each species to deal with sudden increases in substrate concentration.

  11. Bactérias endofíticas no controle e inibição in vitro de Pseudomonas syringae pv tomato, agente da pinta bacteriana do tomateiro Control with endophytic bacteria and in vitro inhibition of Pseudomonas syringae pv tomato, agent of bacterial speck of tomato

    Directory of Open Access Journals (Sweden)

    Juliana Resende Campos Silva

    2008-08-01

    Full Text Available Para avaliar o potencial de 53 isolados de bactérias endofíticas no controle da pinta bacteriana do tomateiro (Lycopersicum esculentum Mill., realizaram-se seleções massais em casa-de-vegetação e a seguir foi avaliado, in vitro, o antagonismo desses isolados sobre a bactéria desafiante Pseudomonas syringae pv. tomato (Pst. A inoculação das bactérias endofíticas foi feita por microbiolização das sementes de tomate cv. Santa Clara e da desafiante (Pst por pulverização. Aos 7, 14 e 21 dias após a inoculação da Pst, foram realizadas as avaliações da severidade da pinta bacteriana, bem como da altura das plantas. As espécies e os isolados bacterianos mais eficazes na redução da severidade da pinta bacteriana foram: Acinetobacter johnsonii (isolado 10, Bacillus pumilus (isolados 3, 12, 20, 39, 51, Paenibacillus macerans (isolados 37 e 47, PIM 11, Bacillus sphaericus (isolado 45, B. amyloliquefaciens (isolado 50, TOM 2, TOM 24 e Staphylococcus aureus (isolado 18. Mais de 50% dos isolados eficazes na redução da severidade foram da espécie Bacillus pumilus. Das espécies endofíticas mais eficazes na redução da severidade da pinta bacteriana, Bacillus pumilus e B. amyloliquefaciens inibiram também o crescimento da Pst in vitro.Vários dos isolados promoveram também o crescimento das plantas.To asses the potential of fifty three isolates of endophytic bacteria on the control of Pseudomonas syringae pv. tomato (Pst in tomato (Lycopersicum esculentum Mill., several screening were done in greenhouse followed by in vitro studies on antagonism of those isolates to Pst. The inoculation of endophytic bacteria was done by microbiolization of tomato cv Santa Clara seeds. The challenging bacterium (Pst inoculation was done by spraying. At 7, 14 and 21 days after Pst inoculation the assessment of bacterial speck severity was done, and height of plants was also measured. The most efficient endophytic species and isolates in reducing

  12. Pseudomonas aeruginosa in Healthcare Settings

    Science.gov (United States)

    ... Sepsis Sharps Safety - CDC Transplant Safety Vaccine Safety Pseudomonas aeruginosa in Healthcare Settings Recommend on Facebook Tweet Share ... aeruginosa . Pseudomonas aeruginosa What types of infections does Pseudomonas aeruginosa cause? Serious Pseudomonas infections usually occur in people ...

  13. Behavioral response of resistant and sensitive Pseudomonas ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-07-18

    Jul 18, 2008 ... Key words: Pseudomonas aeruginosa, cadmium stress, heavy metal resistance. INTRODUCTION. The release of .... plasmids located in the bacterial strains isolated from agricultural and industrial soils ..... esteraromaticum S51 with other strains of non-flocculating sludge bacteria. IWA's Water Environ.

  14. High quality draft genome sequences of Pseudomonas fulva DSM 17717(T), Pseudomonas parafulva DSM 17004(T) and Pseudomonas cremoricolorata DSM 17059(T) type strains.

    Science.gov (United States)

    Peña, Arantxa; Busquets, Antonio; Gomila, Margarita; Mulet, Magdalena; Gomila, Rosa M; Reddy, T B K; Huntemann, Marcel; Pati, Amrita; Ivanova, Natalia; Markowitz, Victor; García-Valdés, Elena; Göker, Markus; Woyke, Tanja; Klenk, Hans-Peter; Kyrpides, Nikos; Lalucat, Jorge

    2016-01-01

    Pseudomonas has the highest number of species out of any genus of Gram-negative bacteria and is phylogenetically divided into several groups. The Pseudomonas putida phylogenetic branch includes at least 13 species of environmental and industrial interest, plant-associated bacteria, insect pathogens, and even some members that have been found in clinical specimens. In the context of the Genomic Encyclopedia of Bacteria and Archaea project, we present the permanent, high-quality draft genomes of the type strains of 3 taxonomically and ecologically closely related species in the Pseudomonas putida phylogenetic branch: Pseudomonas fulva DSM 17717(T), Pseudomonas parafulva DSM 17004(T) and Pseudomonas cremoricolorata DSM 17059(T). All three genomes are comparable in size (4.6-4.9 Mb), with 4,119-4,459 protein-coding genes. Average nucleotide identity based on BLAST comparisons and digital genome-to-genome distance calculations are in good agreement with experimental DNA-DNA hybridization results. The genome sequences presented here will be very helpful in elucidating the taxonomy, phylogeny and evolution of the Pseudomonas putida species complex.

  15. Molecular and chemical dialogues in bacteria-protozoa interactions

    NARCIS (Netherlands)

    Song, Chunxu; Mazzola, M.; Cheng, Xu; Oetjen, Janina; Alexandrov, Theodore; Dorrestein, Pieter C.; Watrous, Jeramie; van der Voort, Menno; Raaijmakers, Jos

    2015-01-01

    Protozoan predation of bacteria can significantly affect soil microbial community composition and ecosystem functioning. Bacteria possess diverse defense strategies to resist or evade protozoan predation. For soil-dwelling Pseudomonas species, several secondary metabolites were proposed to provide

  16. Characterization of a marine-isolated mercury-resistant Pseudomonas putida strain SP1 and its potential application in marine mercury reduction

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Weiwei; Chen, Lingxin; Liu, Dongyan [Chinese Academy of Sciences, Yantai, SD (China). Yantai Inst. of Coastal Zone Research (YICCAS); Chinese Academy of Sciences, Yantai, SD (China). Shandong Provincial Key Lab. of Coastal Zone Environmental Processes

    2012-02-15

    The Pseudomonas putida strain SP1 was isolated from marine environment and was found to be resistant to 280 {mu}M HgCl{sub 2}. SP1 was also highly resistant to other metals, including CdCl{sub 2}, CoCl{sub 2}, CrCl{sub 3}, CuCl{sub 2}, PbCl{sub 2}, and ZnSO{sub 4}, and the antibiotics ampicillin (Ap), kanamycin (Kn), chloramphenicol (Cm), and tetracycline (Tc). mer operon, possessed by most mercury-resistant bacteria, and other diverse types of resistant determinants were all located on the bacterial chromosome. Cold vapor atomic absorption spectrometry and a volatilization test indicated that the isolated P. putida SP1 was able to volatilize almost 100% of the total mercury it was exposed to and could potentially be used for bioremediation in marine environments. The optimal pH for the growth of P. putida SP1 in the presence of HgCl{sub 2} and the removal of HgCl{sub 2} by P. putida SP1 was between 8.0 and 9.0, whereas the optimal pH for the expression of merA, the mercuric reductase enzyme in mer operon that reduces reactive Hg{sup 2+} to volatile and relatively inert monoatomic Hg{sup 0} vapor, was around 5.0. LD50 of P. putida SP1 to flounder and turbot was 1.5 x 10{sup 9} CFU. Biofilm developed by P. putida SP1 was 1- to 3-fold lower than biofilm developed by an aquatic pathogen Pseudomonas fluorescens TSS. The results of this study indicate that P. putida SP1 is a low virulence strain that can potentially be applied in the bioremediation of HgCl{sub 2} contamination over a broad range of pH. (orig.)

  17. Analysis of culture-dependent versus culture-independent techniques for identification of bacteria in clinically obtained bronchoalveolar lavage fluid.

    Science.gov (United States)

    Dickson, Robert P; Erb-Downward, John R; Prescott, Hallie C; Martinez, Fernando J; Curtis, Jeffrey L; Lama, Vibha N; Huffnagle, Gary B

    2014-10-01

    The diagnosis and management of pneumonia are limited by the use of culture-based techniques of microbial identification, which may fail to identify unculturable, fastidious, and metabolically active viable but unculturable bacteria. Novel high-throughput culture-independent techniques hold promise but have not been systematically compared to conventional culture. We analyzed 46 clinically obtained bronchoalveolar lavage (BAL) fluid specimens from symptomatic and asymptomatic lung transplant recipients both by culture (using a clinical microbiology laboratory protocol) and by bacterial 16S rRNA gene pyrosequencing. Bacteria were identified in 44 of 46 (95.7%) BAL fluid specimens by culture-independent sequencing, significantly more than the number of specimens in which bacteria were detected (37 of 46, 80.4%, P ≤ 0.05) or "pathogen" species reported (18 of 46, 39.1%, P ≤ 0.0001) via culture. Identification of bacteria by culture was positively associated with culture-independent indices of infection (total bacterial DNA burden and low bacterial community diversity) (P ≤ 0.01). In BAL fluid specimens with no culture growth, the amount of bacterial DNA was greater than that in reagent and rinse controls, and communities were markedly dominated by select Gammaproteobacteria, notably Escherichia species and Pseudomonas fluorescens. Culture growth above the threshold of 10(4) CFU/ml was correlated with increased bacterial DNA burden (P culture-independent techniques identified a respiratory pathogen missed by culture and clarified whether a cultured "oral flora" species represented a state of acute infection. In summary, we found that bacterial culture of BAL fluid is largely effective in discriminating acute infection from its absence and identified some specific limitations of BAL fluid culture in the diagnosis of pneumonia. We report the first correlation of quantitative BAL fluid culture results with culture-independent evidence of infection. Copyright © 2014

  18. Effect of the temperature of the dipping solution on the antimicrobial effectiveness of various chemical decontaminants against pathogenic and spoilage bacteria on poultry.

    Science.gov (United States)

    Alonso-Hernando, Alicia; Guevara-Franco, José Alfredo; Alonso-Calleja, Carlos; Capita, Rosa

    2013-05-01

    The influence of the temperature of the dipping solution on the antimicrobial effectiveness of several chemical poultry decontaminants was assessed. A total of 765 poultry legs were inoculated with gram-positive bacteria (Listeria monocytogenes, Staphylococcus aureus, Bacillus cereus, or Brochothrix thermosphacta) or gram-negative bacteria (Salmonella enterica serotype Enteritidis, Escherichia coli, Yersinia enterocolitica, or Pseudomonas fluorescens). Samples were dipped for 15 min in solutions (wt/vol) of trisodium phosphate (12%), acidified sodium chlorite (1,200 ppm), citric acid (2%), peroxyacids (220 ppm), chlorine dioxide (50 ppm), or tap water or were left untreated (control). The temperatures of the dipping solutions were 4, 20, or 50°C. Microbiological analyses and pH determinations were carried out after 0, 1, 3, and 5 days of storage at 4°C. In comparison with the control samples, all chemical solutions were effective for reducing microbial loads. The temperature of treatment affected the microbial reductions caused by all chemicals (P peroxyacids were observed at 4°C, all sampling days and microbial groups being considered simultaneously. The highest and the lowest effectiveness for chlorine dioxide were observed at 4 and 50°C, respectively. These results may be of use to meat processors for selecting the best conditions for decontamination treatments and may help the European Regulatory Authorities make their decisions for authorization of poultry decontamination treatments.

  19. Efectos de vinazas sobre bacterias rizosféricas y en la actividad-CO2 y biomasa-C microbiana de un suelo Pachic Haplustoll

    Directory of Open Access Journals (Sweden)

    Miriam Rosero G

    2013-04-01

    Full Text Available En condiciones de casa de malla de la Universidad Nacional de Colombia sede Palmira se estudió los efectos de la aplicación de vinaza, un subproducto de la industria de alcohol carburante, sobre las bacterias rizosféricas Pseudomonas fluorescens y Bacillus subtilis promotoras de crecimiento, la actividad-CO2, biomasa microbiana-C y el cociente metabólico-qCO2 en un suelo Pachic Haplustoll y su relación con el rendimiento de habichuela (Phaseolus vulgaris L.. Se utilizó un diseño completamente al azar con siete tratamientos y cinco repeticiones. Los tratamientos se seleccionaron con base en los requerimientos de K del cultivo (150 kg/ha K2O utilizando como fuentes KCl y vinaza solos y en mezclas. Los tratamientos evaluados y la época de muestreo influyeron (P < 0.05 en la actividad y biomasa microbiana. Los menores valores de estas variables se presentaron en la época de floración del cultivo cuando la demanda de nutrientes es alta. La mezcla en partes iguales de vinaza y KCl favorece la mayor producción de habichuela sin afectar la actividad microbiana; el cociente metabólico indicó estabilidad del sistema en el tiempo y las bacterias rizosféricas presentaron el mejor crecimiento en la mezcla 75% de potasio como vinaza y 25% como KCl.

  20. Bacteria associated with cultures of psathyrella atroumbonata ...

    African Journals Online (AJOL)

    These bacteria include Bacillus licheniformis, Bacillus subtilis, Leuconostoc mesenteroides, Pseudomonas aeruginosa, Bacillus cereus and Staphylococcus aureus. The average bacteria count was 1.0 x 106 cfu/ml and these bacteria grew within pH range of 5.0 and 9.0. the optimum temperature range of growth lied ...

  1. Transcriptomic profiling of microbe-microbe interactions reveals the specific response of the biocontrol strain P. fluorescens In5 to the phytopathogen Rhizoctonia solani.

    Science.gov (United States)

    Hennessy, Rosanna C; Glaring, Mikkel A; Olsson, Stefan; Stougaard, Peter

    2017-08-10

    Few studies to date report the transcriptional response of biocontrol bacteria toward phytopathogens. In order to gain insights into the potential mechanism underlying the antagonism of the antimicrobial producing strain P. fluorescens In5 against the phytopathogens Rhizoctonia solani and Pythium aphanidermatum, global RNA sequencing was performed. Differential gene expression profiling of P. fluorescens In5 in response to either R. solani or P. aphanidermatum was investigated using transcriptome sequencing (RNA-seq). Total RNA was isolated from single bacterial cultures of P. fluorescens In5 or bacterial cultures in dual-culture for 48 h with each pathogen in biological triplicates. RNA-seq libraries were constructed following a default Illumina stranded RNA protocol including rRNA depletion and were sequenced 2 × 100 bases on Illumina HiSeq generating approximately 10 million reads per sample. No significant changes in global gene expression were recorded during dual-culture of P. fluorescens In5 with any of the two pathogens but rather each pathogen appeared to induce expression of a specific set of genes. A particularly strong transcriptional response to R. solani was observed and notably several genes possibly associated with secondary metabolite detoxification and metabolism were highly upregulated in response to the fungus. A total of 23 genes were significantly upregulated and seven genes were significantly downregulated with at least respectively a threefold change in expression level in response to R. solani compared to the no fungus control. In contrast, only one gene was significantly upregulated over threefold and three transcripts were significantly downregulated over threefold in response to P. aphanidermatum. Genes known to be involved in synthesis of secondary metabolites, e.g. non-ribosomal synthetases and hydrogen cyanide were not differentially expressed at the time points studied. This study demonstrates that genes possibly involved in

  2. Pseudomonas spp. convert metmyoglobin into deoxymyoglobin.

    Science.gov (United States)

    Motoyama, Michiyo; Kobayashi, Miho; Sasaki, Keisuke; Nomura, Masaru; Mitsumoto, Mitsuru

    2010-01-01

    Meat 'reddening' by bacteria was observed in chilled beef. To identify the reddening bacteria, isolates were inoculated onto beef and the changes in CIE L*a*b* values monitored. As a result, two Pseudomonas spp., including Pseudomonas fragi which is commonly observed in raw meat, were selected and identified as reddening bacteria. The reddening was coincidentally occurred with the appearance of slime, and the increase in thiobarbituric acid-reactive substances (TBARS) was simultaneously suppressed. In myoglobin-containing nutrient broth, it is shown spectroscopically that P. fragi converted metmyoglobin into deoxymyoglobin. It was concluded that the meat reddening was due to the formation of deoxymyoglobin, induced by the very-low-oxygen tension brought about by Pseudomonad's oxygen consumption: This oxygen depletion simultaneously suppressed TBARS increase.

  3. Growth of Pseudomonas spp. in cottage cheese

    DEFF Research Database (Denmark)

    Østergaard, Nina Bjerre; Dalgaard, Paw

    of spoilage microorganisms in cottage cheese can cause undesirable alterations in flavour, odour, appearance and texture. Contamination and growth of psychrotolerant pseudomonads including Pseudomonas fragi and Pseudomonas putida has been reported for cottage cheese but the influence of these bacteria...... on product spoilage and shelf-life remains poorly described. The present study used a quantitative microbial ecology approach to model and predict the effect of product characteristics and storage conditions on growth of psychrotolerant pseudomonads in cottage cheese. The effect of temperature (5-15˚C) and p...

  4. Behaviour of co-inoculated pathogenic and spoilage bacteria on poultry following several decontamination treatments.

    Science.gov (United States)

    Alonso-Hernando, Alicia; Capita, Rosa; Alonso-Calleja, Carlos

    2012-10-01

    The potential of chemical decontaminants to cause harmful effects on human health is among the causes of the rejection of antimicrobial treatments for removing surface contamination from poultry carcasses in the European Union. This study was undertaken to determine whether decontaminants might give a competitive advantage to pathogenic bacteria on poultry and involve a potential risk to consumer. A total of 144 chicken legs were co-inoculated with similar concentrations of pathogenic bacteria (Listeria monocytogenes, Staphylococcus aureus, Salmonella enterica serotype Enteritidis or Escherichia coli) and spoilage bacteria (Brochothrix thermosphacta or Pseudomonas fluorescens). Samples were dipped for 15min in solutions (w/v) of trisodium phosphate (12%; TSP), acidified sodium chlorite (1200ppm; ASC), citric acid (2%; CA), peroxyacids (220ppm; PA) or chlorine dioxide (50ppm; CD), or were left untreated (control). Microbiological analyses were carried out on day 0 and every 24h until day 7 of storage (at 10±1°C). The modified Gompertz equation was used as the primary model to fit observed data. TSP, ASC and CA were effective in extending the lag phase (L, ranging from 1.47±1.34days to 4.06±1.16days) and in decreasing the concentration of bacteria during the stationary phase (D, ranging from 2.46±0.51 log(10) cfu/cm(2) to 8.64±0.53 log(10) cfu/cm(2)), relative to the control samples (L values ranging from 0.59±0.38days and 2.52±2.28days, and D values ranging from 6.32±0.89 log(10) cfu/cm(2) to 9.39±0.39 log(10) cfu/cm(2), respectively). Both on untreated and on most decontaminated samples the overgrowth of spoilage bacteria among the species tested was observed throughout storage, suggesting that spoilage would occur prior to any noteworthy increase in the levels of pathogenic microorganisms. However, L. monocytogenes counts similar to, or higher than, those for spoilage bacteria were observed on samples treated with TSP, ASC or CA, suggesting that these

  5. Growth of non-Campylobacter, oxidase-positive bacteria on selective Campylobacter agar.

    OpenAIRE

    Moskowitz, L B; Chester, B

    1982-01-01

    A total of 67 oxidase-positive, gram-negative bacteria were tested for growth on selective Campylobacter agar (Blaser formulation, BBL Microbiology Systems, Cockeysville, Md.) at 42 degrees C under microaerophilic conditions. Although the growth of most of these bacteria was prevented, all strains of Achromobacter xylosoxidans, Pseudomonas aeruginosa, Pseudomonas putrefaciens, Pseudomonas alcaligenes, and Pseudomonas pseudoalcaligenes grew as well as Campylobacter fetus subsp. jejuni.

  6. Phylogenomics and systematics in Pseudomonas

    Directory of Open Access Journals (Sweden)

    Margarita eGomila

    2015-03-01

    Full Text Available The genus Pseudomonas currently contains 144 species, making it the genus of Gram-negative bacteria that contains the largest number of species. Currently, multilocus sequence analysis (MLSA is the preferred method for establishing the phylogeny between species and genera. Four partial gene sequences of housekeeping genes (16S rRNA, gyrB, rpoB and rpoD were obtained from 112 complete or draft genomes of strains related to the genus Pseudomonas that were available in databases. These genes were analyzed together with the corresponding sequences of 133 Pseudomonas type strains of validly published species to assess their correct phylogenetic assignations. We confirmed that 30% of the sequenced genomes of non-type strains were not correctly assigned at the species level in the accepted taxonomy of the genus and that 20% of the strains were not identified at the species level. Most of these strains had been isolated and classified several years ago, and their taxonomic status has not been updated by modern techniques. MLSA was also compared with indices based on the analysis of whole-genome sequences that have been proposed for species delineation, such as tetranucleotide usage patterns (TETRA, average nucleotide identity (ANIm, based on MUMmer and ANIb, based on BLAST and genome-to-genome distance (GGDC. TETRA was useful for discriminating Pseudomonas from other genera, whereas ANIb and GGDC clearly separated strains of different species. ANIb showed the strongest correlation with MLSA. The correct species classification is a prerequisite for most diversity and evolutionary studies. This work highlights the necessity for complete genomic sequences of type strains to build a phylogenomic taxonomy and that all new genome sequences submitted to databases should be correctly assigned to species to avoid taxonomic inconsistencies.

  7. Contagem, isolamento e caracterização de bactérias psicrotróficas contaminantes de leite cru refrigerado Counting, isolation and characterization of psychrotrophic bacteria from refrigerated raw milk

    Directory of Open Access Journals (Sweden)

    Edna Froeder Arcuri

    2008-11-01

    Full Text Available Com os objetivos de quantificar, isolar e caracterizar bactérias psicrotróficas contaminantes de leite cru refrigerado, produzido na região da Zona da Mata de Minas Gerais e Sudeste do Rio de Janeiro, foram analisadas amostras de leite coletadas de 20 tanques coletivos e 23 tanques individuais. As contagens de bactérias psicrotróficas nas amostras de leite para os dois tipos de tanques de refrigeração variaram entre 10² e 10(7 Unidades Formadoras de Colônias (UFC ml-1, porém, um maior número de tanques coletivos apresentou contagens acima de 1 x 10(5 UFC ml-1. Foi verificada a predominância de bactérias psicrotróficas gram-negativas (81,2%, que foram identificadas pelos sistemas API 20E e API 20NE nos gêneros: Aeromonas, Alcaligenes, Acinetobacter, Burkholderia,Chryseomonas, Enterobacter, Ewingella, Klebsiella, Hafnia, Methylobacterium, Moraxella, Pantoea, Pseudomonas, Serratia, Sphingomonas e Yersinia. As bactérias gram-positivas (18,8% foram identificadas com API 50 CH, API Coryne e API Staph, nos gêneros: Bacillus, Brevibacterium, Cellum/Microbacterium, Kurthia e Staphylococcus. Os sistemas API utilizados não identificaram todos os isolados bacterianos. Pseudomonas foi o gênero mais isolado e P. fluorescens foi a espécie predominante. A maioria dos isolados bacterianos apresentou atividade proteolítica e/ou lipolítica a temperaturas de refrigeração de 4°C, 7°C e 10°C, evidenciando seu alto potencial de deterioração do leite e dos produtos lácteos. Os resultados ressaltam que maior atenção deve ser dada aos procedimentos que impeçam a contaminação do leite por esses microrganismos.This study aimed to quantify, isolate and characterize psychrotrophic bacteria from refrigerated raw milk produced at the ‘Mata’ Region of Minas Gerais State and Southeast of Rio de Janeiro State, Brazil. Raw milk samples, were collected at the farms, from 20 collective refrigerated tanks and 23 individual refrigerated tanks

  8. A novel nanoparticle approach