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Sample records for leguminosarum biovar phaseoli

  1. Interacción de Tsukamurella paurometabola C-924 con Rhizobium leguminosarum biovar phaseoli CFH en el cultivo de frijol

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    Marieta Marín Bruzos

    2013-01-01

    Full Text Available En el estudio se evaluaron, mediante análisis de los parámetros fenológicos de las plantas, la interacción de Tsukamurella paurometabola C-924 con Rhizobium leguminosarum biovar phaseoli CFH en el cultivo de frijol. Se emplearon cuatro tratamientos: plantas sin inocular (control, inoculadas con T. paurometabola C-924, inoculadas con R. leguminosarum biovar phaseoli CFH e inoculadas con ambas cepas. Se observaron diferencias significativas (P < 0.01 en los porcentajes de germinación de las plantas tratadas con microorganismos de forma independiente o conjunta con respecto al control sin inocular. Se determinó que la inoculación de T. paurometabola C-924 afectó el proceso de nodulación de R. leguminosarum biovar phaseoli CFH. Sin embargo, esto no incidió de manera significativa en la altura de las plantas ni en el diámetro del tallo, ya que no se encontraron diferencias entre los tratamientos para estos parámetros. Para el número de hojas, los mejores resultados se obtuvieron con la aplicación de T. paurometabola C-924. Se concluyó que la interacción de T. paurometabola C-924 con R. leguminosarum biovar phaseoli CFH en el frijol estimuló significativamente la germinación de las semillas y el número de hojas de las plantas con respecto al control sin inocular. Aunque la aplicación de T. paurometabola C-924 no favoreció la nodulación de R. leguminosarum biovar phaseoli CFH, esto no afectó las características fenológicas del cultivo.

  2. Mutation of a Broadly Conserved Operon (RL3499-RL3502) from Rhizobium leguminosarum Biovar viciae Causes Defects in Cell Morphology and Envelope Integrity▿†

    Science.gov (United States)

    Vanderlinde, Elizabeth M.; Magnus, Samantha A.; Tambalo, Dinah D.; Koval, Susan F.; Yost, Christopher K.

    2011-01-01

    The bacterial cell envelope is of critical importance to the function and survival of the cell; it acts as a barrier against harmful toxins while allowing the flow of nutrients into the cell. It also serves as a point of physical contact between a bacterial cell and its host. Hence, the cell envelope of Rhizobium leguminosarum is critical to cell survival under both free-living and symbiotic conditions. Transposon mutagenesis of R. leguminosarum strain 3841 followed by a screen to isolate mutants with defective cell envelopes led to the identification of a novel conserved operon (RL3499-RL3502) consisting of a putative moxR-like AAA+ ATPase, a hypothetical protein with a domain of unknown function (designated domain of unknown function 58), and two hypothetical transmembrane proteins. Mutation of genes within this operon resulted in increased sensitivity to membrane-disruptive agents such as detergents, hydrophobic antibiotics, and alkaline pH. On minimal media, the mutants retain their rod shape but are roughly 3 times larger than the wild type. On media containing glycine or peptides such as yeast extract, the mutants form large, distorted spheres and are incapable of sustained growth under these culture conditions. Expression of the operon is maximal during the stationary phase of growth and is reduced in a chvG mutant, indicating a role for this sensor kinase in regulation of the operon. Our findings provide the first functional insight into these genes of unknown function, suggesting a possible role in cell envelope development in Rhizobium leguminosarum. Given the broad conservation of these genes among the Alphaproteobacteria, the results of this study may also provide insight into the physiological role of these genes in other Alphaproteobacteria, including the animal pathogen Brucella. PMID:21357485

  3. Infection of cattle in Kenya with Brucella abortus biovar 3 and Brucella melitensis biovar 1 genotypes

    NARCIS (Netherlands)

    Muendo, Esther N.; Mbatha, Peter M.; Macharia, Joseph; Abdoel, Theresia H.; Janszen, Paul V.; Pastoor, Rob; Smits, Henk L.

    2012-01-01

    Brucella melitensis biovar 1 was isolated from bovine milk samples from a herd in central Kenya, and Brucella abortus biovar 3 was isolated from aborted fetus materials and vaginal discharge fluids from cattle in central and eastern provinces of Kenya. All infections including those with B.

  4. Symbiotic and phenotypic diversity of Rhizobium leguminosarum bv ...

    African Journals Online (AJOL)

    Administrator

    2011-05-23

    May 23, 2011 ... root nodule bacteria known as Rhizobium leguminosarum bv. viciae. ... nodules, as before by streaking on Yeast-extract mannitol agar medium ... Colony morphology was characterized on YEMA according to. (Ahmed et al.

  5. effects of rhizobuim leguminosarum inoculation on the growth and ...

    African Journals Online (AJOL)

    user

    beneficial symbiotic microorganisms into the plant. Rhizosphere ... Rhizobium strains in the yeast manitol broth were ... inoculants contained sixteen (16) colonies of Rhizobium leguminosarum bacterial cells per milliliter (ml) of the yeast ...

  6. Homology of genes for exopolysaccharide synthesis in Rhizobium leguminosarum and effect of cloned exo genes on nodule formation

    International Nuclear Information System (INIS)

    Skorupska, A.; Derylo, M.

    1993-01-01

    A 5.4 kb BamHI fragment of Rhizobium leguminosarum (R.l.) bv. trifolii TA1 was found to carry genes involved in exopolysaccharide synthesis (exo genes). This fragment was strongly hybridized to the total DNA from DNA from R.l. bv. viciae and bv. phaseoli digested with EcoRI. No homology was found with total DNA of R. melilotic and Rhizobium sp. NGR 234. The exo genes R.l. bv. trifolii TA1 conjugally introduced into R.l. bv. viciae 1302 considerably affected the symbiosis: the nodules induced on vetch were abortive and did not fix nitrogen. On the other hand, Phaseolus beans infected with R.l. bv. trifolii exo genes formed the nitrogen-fixing nodules. It can be conduced that additional copies of exo genes introduced into wild type R.l. strains can disturb the synthesis of acidic exopolysaccharides and affect symbiosis of the plants forming indeterminate nodules, but do not affect symbiosis of the plants forming the determinate nodules. (author). 29 refs, 2 figs, 2 tabs

  7. Synergy between Rhizobium phaseoli and Acidithiobacillus ferrooxidans in the Bioleaching Process of Copper

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    Xuecheng Zheng

    2016-01-01

    Full Text Available This study investigates the synergy of Rhizobium phaseoli and Acidithiobacillus ferrooxidans in the bioleaching process of copper. The results showed that additional R. phaseoli could increase leaching rate and cell number of A. ferrooxidans. When the initial cell number ratio between A. ferrooxidans and R. phaseoli was 2 : 1, A. ferrooxidans attained the highest final cell number of approximately 2 × 108 cells/mL and the highest copper leaching rate of 29%, which is 7% higher than that in the group with A. ferrooxidans only. R. phaseoli may use metabolized polysaccharides from A. ferrooxidans, and organic acids could chelate or precipitate harmful heavy metals to reduce their damage on A. ferrooxidans and promote its growth. Organic acids could also damage the mineral lattice to increase the leaching effect.

  8. Synergy between Rhizobium phaseoli and Acidithiobacillus ferrooxidans in the Bioleaching Process of Copper

    Science.gov (United States)

    Zheng, Xuecheng; Li, Dongwei

    2016-01-01

    This study investigates the synergy of Rhizobium phaseoli and Acidithiobacillus ferrooxidans in the bioleaching process of copper. The results showed that additional R. phaseoli could increase leaching rate and cell number of A. ferrooxidans. When the initial cell number ratio between A. ferrooxidans and R. phaseoli was 2 : 1, A. ferrooxidans attained the highest final cell number of approximately 2 × 108 cells/mL and the highest copper leaching rate of 29%, which is 7% higher than that in the group with A. ferrooxidans only. R. phaseoli may use metabolized polysaccharides from A. ferrooxidans, and organic acids could chelate or precipitate harmful heavy metals to reduce their damage on A. ferrooxidans and promote its growth. Organic acids could also damage the mineral lattice to increase the leaching effect. PMID:26942203

  9. Infection of cattle in Kenya with Brucella abortus biovar 3 and Brucella melitensis biovar 1 genotypes.

    Science.gov (United States)

    Muendo, Esther N; Mbatha, Peter M; Macharia, Joseph; Abdoel, Theresia H; Janszen, Paul V; Pastoor, Rob; Smits, Henk L

    2012-01-01

    Brucella melitensis biovar 1 was isolated from bovine milk samples from a herd in central Kenya, and Brucella abortus biovar 3 was isolated from aborted fetus materials and vaginal discharge fluids from cattle in central and eastern provinces of Kenya. All infections including those with B. melitensis were in cattle with reproductive problems kept in mixed herds indicating that cross infection occurs from small ruminants. Multiple-locus variable-number tandem repeat analysis genotyping revealed a close molecular homology of the B. melitensis isolates with an isolate from Israel and a close homology of the B. abortus isolates with an isolate from Uganda indicating that these genotypes have a wide geographic distribution. Infection of cattle with B. melitensis may complicate the control of brucellosis in this country.

  10. Evaluation of the radioimmunoassay, indirect enzyme linked immunosorbent assay, and dot blot assay for the identification of Xanthomonas campestris pv. phaseoli

    Energy Technology Data Exchange (ETDEWEB)

    Malin, E; Belden, E L; Roth, D

    1985-09-01

    A radioimmunoassay (RIA), an indirect competitive enzyme-linked immunosorbent assay (ELISA), and a dot-blot modification of the ELISA were evaluated for detection and identification of Xanthomonas campestris pv. phaseoli (X. c. pv. phaseoli). RIA and the dot blot tests were specific for X. c. pv. phaseoli; however, significant cross reactions occurred in the indirect competitive ELISA when using anti-X. c. pv. phaseoli antiserum against other closely related bacteria. The sensitivity level of all procedures for X. c. pv. phaseoli was approximately l0/sup 5/ colony forming unitsmL. All procedures were unsatisfactory in reliably detecting low levels of X. c. pv. phaseoli directly from extracts of bean seed. However when used in conjunction with ilution plating the dot blot assay and the RIA would be useful in specifically identifying X. c. pv. phaseoli. The relative merits of these tests for identification of X. c. pv. phaseoli are discussed.

  11. Investigation of Ureaplasma urealyticum biovars and their relationship with antimicrobial resistance

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    Zhu Chang-tai

    2011-01-01

    Full Text Available Purpose: To develop Taqman fluorescence quantitative polymerase chain reaction (PCR method for investigating the characteristics of the distributions of Ureaplasma urealyticum (UU biovars and to explore the relationship between UU biovars and antimicrobial resistance. Materials and Methods: By the method of culture, Ureaplasma species were detected. Taqman fluorescence quantitative PCR for detecting UU biovars were developed and UU clinical isolates were detected to distinguish biovars. The broth micro-dilution susceptibility testing methods were used to determine UU susceptibility. Results: By Taqman PCR method, UU biovars was successfully detected. Of 126 samples, biovar 1 was found in 73 (57.94%. There was a statistical difference between genital-urinary tract infection group and asymptomatic group (P<0.05. In the region, UU biovar 1 to 9 kinds of agents kept higher susceptibility rates, but biovar 2 maintained higher susceptibility rates only to tetracyclines. Compared with biovar 1, UU biovar 2 resistance rates to 7 kinds of agents were higher (P<0.05. Conclusions: (1 Our new established Taqman PCR method is a useful tool for screening UU biovars. (2 UU biovar 1 predominated in asymptomatic population; whereas in genital-urinary tract infection population UU biovar 2 had a higher proportion. (3 The characteristics of drug resistance were different between UU biovars. Overall, both two biovars remained higher susceptibility rates to tetracyclines. A majority of biovor 1 strains were sensitive to macrolides and quinolones; while only a small number of biovar 2 strains kept sensitive to roxithromycin and quinolones, a large proportion of biovar 2 strains were found in intermediate ranges.

  12. Interaction of Acidithiobacillus ferrooxidans, Rhizobium phaseoli and Rhodotorula sp. in bioleaching process based on Lotka–Volterra model

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    Xuecheng Zheng

    2016-07-01

    Conclusion: The relationship among microorganisms during leaching could be described appropriately by Lotka–Volterra model between the initial and peak values. The relationship of A. ferrooxidans and R. phaseoli could be considered as mutualism, whereas, the relationship of A. ferrooxidans and R. phaseoli could be considered as commensalism.

  13. Characterization and functional analysis of seven flagellin genes in Rhizobium leguminosarum bv. viciae. Characterization of R. leguminosarum flagellins

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    Tambalo Dinah D

    2010-08-01

    Full Text Available Abstract Background Rhizobium leguminosarum bv. viciae establishes symbiotic nitrogen fixing partnerships with plant species belonging to the Tribe Vicieae, which includes the genera Vicia, Lathyrus, Pisum and Lens. Motility and chemotaxis are important in the ecology of R. leguminosarum to provide a competitive advantage during the early steps of nodulation, but the mechanisms of motility and flagellar assembly remain poorly studied. This paper addresses the role of the seven flagellin genes in producing a functional flagellum. Results R. leguminosarum strains 3841 and VF39SM have seven flagellin genes (flaA, flaB, flaC, flaD, flaE, flaH, and flaG, which are transcribed separately. The predicted flagellins of 3841 are highly similar or identical to the corresponding flagellins in VF39SM. flaA, flaB, flaC, and flaD are in tandem array and are located in the main flagellar gene cluster. flaH and flaG are located outside of the flagellar/motility region while flaE is plasmid-borne. Five flagellin subunits (FlaA, FlaB, FlaC, FlaE, and FlaG are highly similar to each other, whereas FlaD and FlaH are more distantly related. All flagellins exhibit conserved amino acid residues at the N- and C-terminal ends and are variable in the central regions. Strain 3841 has 1-3 plain subpolar flagella while strain VF39SM exhibits 4-7 plain peritrichous flagella. Three flagellins (FlaA/B/C and five flagellins (FlaA/B/C/E/G were detected by mass spectrometry in the flagellar filaments of strains 3841 and VF39SM, respectively. Mutation of flaA resulted in non-motile VF39SM and extremely reduced motility in 3841. Individual mutations of flaB and flaC resulted in shorter flagellar filaments and consequently reduced swimming and swarming motility for both strains. Mutant VF39SM strains carrying individual mutations in flaD, flaE, flaH, and flaG were not significantly affected in motility and filament morphology. The flagellar filament and the motility of 3841 strains

  14. Identification of Glomerella cingulata f. sp phaseoli recombinants by RAPD markers.

    Science.gov (United States)

    Camargo, O A; Souza, E A; Mendes-Costa, M C; Santos, J B; Soares, M A

    2007-09-30

    We examined the capacity of strains of Glomerella cingulata f. sp phaseoli fungus (Colletotrichum lindemuthianum sexual stage) to form recombinants, using random amplified polymorphic DNA (RAPD). Crosses of all possible combinations between strains 40, 42, 20, 21, 22, 23, 24, 25, and 26 were made on Petri dishes using M3 culture medium. The 42 x 21 cross produced the largest number of perithecia and five asci; the respective ascospores were isolated. RAPD analysis was performed on the parents and descendants. The 62 polymorphic RAPD bands obtained were used to assess the genetic similarity using the method of Sorence and Dice and clustering analysis in the form of a dendrogram by the UPGMA method. The RAPD markers allowed identification of recombinants from the cross between strains 42 and 21 of G. cingulata f. sp phaseoli and 40 ascospores presented 63 and 49% genetic similarity with parents 2 (strain 42) and 1 (strain 21), respectively.

  15. ASSESSMENT OF Trichoderma ISOLATES FOR VIRULENCE EFFICACY ON Fusarium oxysporum F. sp. Phaseoli

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    Jane Otadoh

    2010-10-01

    Full Text Available Trichoderma has been widely studied for their biocontrol ability, but their use as biocontrol agents in agriculture is limited due to the unpredictable efficiency which is affected by biotic and abiotic factors in soil. Isolates of Trichoderma from Embu soils were evaluated for their ability to control Fusarium oxysporum f. sp. phaseoli., in vitro and promote seedling growth in the greenhouse. Bioassays were run using dual cultures and diffusible compound production analysis. The Trichoderma isolates significantly (p

  16. Ultrastructural aspects in perithecia hyphae septal pores of Glomerella cingulata F. SP. Phaseoli

    OpenAIRE

    Roca M.,María Gabriela; Ongarelli,Maria das Graças; Davide,Lisete Chamma; Mendes-Costa,Maria Cristina

    2000-01-01

    Glomerella cingulata (Stonem.) Spauld. & Schrenk f. sp. phaseoli, better known in its anamorphic state Colletotrichum lindemuthianum (Sacc. & Magn.) Briosi & Cav., is a causal agent of anthracnose in beans (Phaseolus vulgaris L.). Ultrastructural aspects of the perithecial hyphae of this pathogen were studied. The perithecia hyphae septal pores were found either plugged by a vesicle or unplugged. Some perithecia hyphae septa presented no pore. The Woronin bodies, close to the sept...

  17. Identification at Biovar Level of Brucella Isolates Causing Abortion in Small Ruminants of Iran

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    Ali Mohammad Behroozikhah

    2012-01-01

    Full Text Available To determine the most prevalent biovar responsible for brucellosis in sheep and goat populations of Iran, a cross-sectional study was carried out over 2 years in six provinces selected based on geography and disease prevalence. Specimens obtained from referred aborted sheep and goat fetuses were cultured on Brucella selective media for microbiological isolation. Brucellae were isolated from 265 fetuses and examined for biovar identification using standard microbiological methods. Results showed that 246 isolates (92.8% were B. melitensis biovar 1, 18 isolates (6.8% were B. melitensis biovar 2, and, interestingly, one isolate (0.4% obtained from Mazandaran province was B. abortus biovar 3. In this study, B. melitensis biovar 3 was isolated in none of the selected provinces, and all isolates from 3 provinces (i.e., Chehar-mahal Bakhtiari, Markazi, and Ilam were identified only as B. melitensis biovar 1. In conclusion, we found that B. melitensis biovar 1 remains the most prevalent cause of small ruminant brucellosis in various provinces of Iran.

  18. The ethylene-inhibitor aminoethoxyvinylglycine restores normal nodulation by Rhizobium leguminosarum biovar. viciae on Vicia sativa subsp. nigra by suppressing the 'Thick and short roots' phenotype

    NARCIS (Netherlands)

    Zaat, S. A.; van Brussel, A. A.; Tak, T.; Lugtenberg, B. J.; KIJNE, J. W.

    1989-01-01

    Nodulation of Vicia sativa subsp. nigra L. by Rhizobium bacteria is coupled to the development of thick and short roots (Tsr). This root phenotype as well as root-hair induction (Hai) and root-hair deformation (Had) are caused by a factor(s) produced by the bacteria in response to plant flavonoids.

  19. The ethylene-inhibitor aminoethoxyvinylglycine restores normal nodulation by Rhizobium leguminosarum biovar. viciae on Vicia sativa subsp. nigra by suppressing the 'Thick and short roots' phenotype.

    Science.gov (United States)

    Zaat, S A; Van Brussel, A A; Tak, T; Lugtenberg, B J; Kijne, J W

    1989-02-01

    Nodulation of Vicia sativa subsp. nigra L. by Rhizobium bacteria is coupled to the development of thick and short roots (Tsr). This root phenotype as well as root-hair induction (Hai) and root-hair deformation (Had) are caused by a factor(s) produced by the bacteria in response to plant flavonoids. When very low inoculum concentrations (0.5-5 bacteria·ml(-1)) were used, V. sativa plants did not develop the Tsr phenotype and became nodulated earlier than plants with Tsr roots. Furthermore, the nodules of these plants were located on the primary root in contrast to nodules on Tsr roots, which were all located at sites of lateral-root emergence. The average numbers of nodules per plant were not significantly different for these two types of nodulation. Root-growth inhibition and Hai, but not Had, could be mimicked by ethephon, and inhibited by aminoethoxyvinylglycine (AVG). Addition of AVG to co-cultures of Vicia sativa and the standard inoculum concentration of 5·10(5) bacteria·ml(-1) suppressed the development of the Tsr phenotype and restored nodulation to the pattern that was observed with very low concentrations of bacteria (0.5-5 bacteria·ml(-1)). The delay in nodulation on Tsr roots appeared to be caused by the fact that nodule meristems did not develop on the primary root, but only on the emerging laterals. The relationship between Tsr, Hai, Had, and nodulation is discussed.

  20. Visualization of Nodulation Gene Activity on the Early Stages of Rhizobium leguminosarum bv. viciae Symbiosis

    Czech Academy of Sciences Publication Activity Database

    Chovanec, Pavel; Novák, Karel

    2005-01-01

    Roč. 50, č. 4 (2005), s. 323-331 ISSN 0015-5632 R&D Projects: GA ČR GA521/03/0192 Institutional research plan: CEZ:AV0Z50200510 Keywords : nodulation * rhizobium leguminosarum * vicia tetrasperma Subject RIV: EE - Microbiology, Virology Impact factor: 0.918, year: 2005

  1. Rhizoplane colonisation of peas by Rhizobium leguminosarum bv. viceae and a deleterious Pseudomonas putida

    NARCIS (Netherlands)

    Berggren, I.; Alstrom, S.; Vuurde, van J.W.L.; Martensson, A.M.

    2005-01-01

    Pseudomonas putida strain angstrom 313, a deleterious rhizosphere bacterium, reduced pea nitrogen content when inoculated alone or in combination with Rhizobium leguminosarum bv. viceae on plants in the presence of soil under greenhouse conditions. When plants were grown gnotobiotically in liquid

  2. Studies on the expression of plasmid-borne genes in the endosymbiotic state of Rhizobium leguminosarum

    NARCIS (Netherlands)

    Krol, A.J.M.

    1982-01-01

    The subject matter of the research reported in this thesis is the role of plasmid-borne genes of Rhizobium in symbiosis and nitrogen fixation. Plasmid DNA was isolated from Rhizobium leguminosarum strain PRE and the expression of plasmid DNA in nitrogen

  3. Energy supply for dinitrogen fixation by Azotobacter vinelandii and by bacteroids of Rhizobium leguminosarum

    NARCIS (Netherlands)

    Laane, N.C.M.

    1980-01-01

    The central issue of this thesis is how obligate aerobes, such as Rhizobium leguminosarum bacteroids and Azotobacter vinelandii, generate and regulate the energy supply (in the form of ATP and reducing equivalents) for nitrogenase.
    In an effective

  4. Ultrastructural aspects in perithecia hyphae septal pores of Glomerella cingulata F. SP. Phaseoli

    Directory of Open Access Journals (Sweden)

    Roca M. María Gabriela

    2000-01-01

    Full Text Available Glomerella cingulata (Stonem. Spauld. & Schrenk f. sp. phaseoli, better known in its anamorphic state Colletotrichum lindemuthianum (Sacc. & Magn. Briosi & Cav., is a causal agent of anthracnose in beans (Phaseolus vulgaris L.. Ultrastructural aspects of the perithecial hyphae of this pathogen were studied. The perithecia hyphae septal pores were found either plugged by a vesicle or unplugged. Some perithecia hyphae septa presented no pore. The Woronin bodies, close to the septal pores, appeared as globose structures which were more electron dense than the occlusions plugging the septal pore.

  5. Dimorfismo de ascósporos em Glomerella cingulata f.sp. phaseoli

    OpenAIRE

    Castro, Renata A.; Mendes-Costa, Maria C.; Souza, Elaine A.

    2006-01-01

    O objetivo deste trabalho foi caracterizar linhagens de Glomerella cingulata f. sp. phaseoli quanto ao crescimento micelial, formação de peritécio e dimorfismo de ascósporos. Foram avaliadas quatro linhagens nos meios de cultura de folhas de feijoeiro e meio M3 e em condições com e sem fotoperíodo. O comprimento dos ascósporos da linhagem normal e da mutante foi determinado. Houve diferença significativa quanto ao crescimento micelial e tamanho dos ascósporos. Constatou-se, pela primeira vez,...

  6. Clonal outbreaks of [ Pasteurella] pneumotropica biovar Heyl in two mouse colonies

    DEFF Research Database (Denmark)

    Adhikary, Sadhana; Bisgaard, Magne; Dagnæs-Hansen, Frederik

    2017-01-01

    The aim of this study was to document the pathogenic role of biovar Heyl of [Pasteurella] pneumotropica in mouse colonies. Fifty-three isolates associated with mastitis and orbital, cutaneous and vaginal abscesses as well as isolates from the nose and vagina of healthy mice were investigated...... strains with the same rpoB sequence type, as shown by the PFGE profiles. The investigation documented that members of biovar Heyl of [P.] pneumotropica caused disease outbreaks in mouse colonies since the clonality indicated a primary role of [P.] pneumotropica biovar Heyl in the infections observed....

  7. A DNA-Based Procedure for In Planta Detection of Fusarium oxysporum f. sp. phaseoli.

    Science.gov (United States)

    Alves-Santos, Fernando M; Ramos, Brisa; García-Sánchez, M Asunción; Eslava, Arturo P; Díaz-Mínguez, José María

    2002-03-01

    ABSTRACT We have characterized strains of Fusarium oxysporum from common bean fields in Spain that were nonpathogenic on common bean, as well as F. oxysporum strains (F. oxysporum f. sp. phaseoli) pathogenic to common bean by random amplified polymorphic DNA (RAPD) analysis. We identified a RAPD marker (RAPD 4.12) specific for the highly virulent pathogenic strains of the seven races of F. oxysporum f. sp. phaseoli. Sequence analysis of RAPD 4.12 allowed the design of oligonucleotides that amplify a 609-bp sequence characterized amplified region (SCAR) marker (SCAR-B310A280). Under controlled environmental and greenhouse conditions, detection of the pathogen by polymerase chain reaction was 100% successful in root samples of infected but still symptomless plants and in stem samples of plants with disease severity of >/=4 in the Centro Internacional de Agricultura Tropical (CIAT; Cali, Colombia) scale. The diagnostic procedure can be completed in 5 h and allows the detection of all known races of the pathogen in plant samples at early stages of the disease with no visible symptoms.

  8. Reaction of common bean lines to Xanthomonas axonopodis pv. phaseoli and Curtobacterium flaccumfaciens pv. flaccumfaciens

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    Tamires Ribeiro

    2016-12-01

    Full Text Available The aim of this study was to evaluate the resistance of 58 common bean lines against common bacterial blight (Xanthomonas axonopodis pv. phaseoli and bacterial wilt (Curtobacterium flaccumfaciens pv. flaccumfaciens. The experimental design consisted of completely randomized blocks, with four replications per pathogen. The results were subjected to variance analysis by the F test at 1% probability. Significant differences between the treatments indicated different resistance levels among the lines against both pathogens. According to the Scott-Knott test, six lines were resistant to Xanthomonas axonopodis pv. phaseoli, 14 moderately resistant, and 38 susceptible. To Curtobacterium flaccumfaciens pv. flaccumfaciens, 11 lines were resistant, 26 moderately resistant and 21 susceptible. Among these, the lines Pr10-3-4/1, Pr10-5-2/1 and Pr10-5- 2/2 of the black bean group and C10-2-4/2 of the Carioca group were resistant to both major bacterial diseases affecting common bean in Brazil.

  9. The center for plant and microbial complex carbohydrates at the University of Georgia Complex Carbohydrate Research Center

    Energy Technology Data Exchange (ETDEWEB)

    Albersheim, P.; Darvill, A.

    1991-08-01

    Research from the Complex Carbohydrates Research Center at the University of Georgia is presented. Topics include: Structural determination of soybean isoflavones which specifically induce Bradyrhizobium japonicum nodD1 but not the nodYABCSUIJ operon; structural analysis of the lipopolysaccharides (LPSs) from symbiotic mutants of Bradyrhizobium japonicum; structural characterization of lipooligosaccharides from Bradyrhizobium japonicum that are required for the specific nodulation of soybean; structural characterization of the LPSs from R. Leguminosarum biovar phaseoli, the symbiont of bean; characterization of bacteroid-specific LPS epitopes in R. leguminosarum biovar viciae; analysis of the surface polysaccharides of Rhizobium meliloti mutants whose lipopolysaccharides and extracellular polysaccharides can have the same function in symbiosis; characterization of a polysaccharide produced by certain Bradyrhizobium japonicum strains within soybean nodules; structural analysis of a streptococcal adhesin polysaccharide receptor; conformational studies of xyloglucan, the role of the fucosylated side chain in surface-specific cellulose-xyloglucan interactions; the structure of an acylated glucosamine oligosaccharide signal molecule (nod factor) involved in the symbiosis of Rhizobium leguminosarum biovar viciae with its host Vicia sativa; investigating membrane responses induced by oligogalacturonides in cultured cells; the polygalacturonase inhibitor protein; characterization of the self-incompatability glycoproteins from Petunia hybrida; investigation of the cell wall polysaccharide structures of Arabidopsis thaliana; and the glucan inhibition of virus infection of tabacco.

  10. The center for plant and microbial complex carbohydrates at the University of Georgia Complex Carbohydrate Research Center. Annual report, September 15, 1990--December 31, 1991

    Energy Technology Data Exchange (ETDEWEB)

    Albersheim, P.; Darvill, A.

    1991-08-01

    Research from the Complex Carbohydrates Research Center at the University of Georgia is presented. Topics include: Structural determination of soybean isoflavones which specifically induce Bradyrhizobium japonicum nodD1 but not the nodYABCSUIJ operon; structural analysis of the lipopolysaccharides (LPSs) from symbiotic mutants of Bradyrhizobium japonicum; structural characterization of lipooligosaccharides from Bradyrhizobium japonicum that are required for the specific nodulation of soybean; structural characterization of the LPSs from R. Leguminosarum biovar phaseoli, the symbiont of bean; characterization of bacteroid-specific LPS epitopes in R. leguminosarum biovar viciae; analysis of the surface polysaccharides of Rhizobium meliloti mutants whose lipopolysaccharides and extracellular polysaccharides can have the same function in symbiosis; characterization of a polysaccharide produced by certain Bradyrhizobium japonicum strains within soybean nodules; structural analysis of a streptococcal adhesin polysaccharide receptor; conformational studies of xyloglucan, the role of the fucosylated side chain in surface-specific cellulose-xyloglucan interactions; the structure of an acylated glucosamine oligosaccharide signal molecule (nod factor) involved in the symbiosis of Rhizobium leguminosarum biovar viciae with its host Vicia sativa; investigating membrane responses induced by oligogalacturonides in cultured cells; the polygalacturonase inhibitor protein; characterization of the self-incompatability glycoproteins from Petunia hybrida; investigation of the cell wall polysaccharide structures of Arabidopsis thaliana; and the glucan inhibition of virus infection of tabacco.

  11. Biological control of Fusarium solani f. sp. phaseoli the causal agent of root rot of bean using Bacillus subtilis CA32 and Trichoderma harzianum RU01

    Directory of Open Access Journals (Sweden)

    Saman Abeysinghe

    2007-09-01

    Full Text Available Root rot, caused by Fusarium solani f. sp. phaseoli, is one of the main root diseases impacting production of common bean in Sri Lanka. Rhizobacteria were screened in dual Petri plate assays to select antagonistic strains against F. solani f. sp. phaseoli. B. subtilis CA32 effectively antagonized the pathogen. T. harzianum RU01 also showed the antagonistic activity. The efficacy of the B. subtilis CA32 and the T. harzianum RU01 were tested in greenhouse pot experiments against F. solani f. sp. phaseoli. Seed bacterization with B. subtilis CA32 and T. harzianum RU01 significantly protected bean seedlings from F. solani f. sp. phaseoli compared to the untreated control plants. Plant protection was more pronounced in T. harzianum RU01 treated plants than bacterized plants. Enhanced root growth was observed only T. harzianum RU01 treated plants, suggesting that the biotic modifications of the mycorrhizosphere as a result of colonization with T. harzianum RU01.

  12. Ralstonia solanacearum biovar 1 associated with a new outbreak of potato brown rot in Portugal

    Directory of Open Access Journals (Sweden)

    Leonor Cruz

    2008-10-01

    Full Text Available In May 2007, potato plants exhibiting symptoms possibly of brown rot were collected in some potato fields in the Baixo Mondego region (Center, Portugal, as a part of a nationwide programme to monitor Ralstonia solanacearum. All laboratory procedures laid down in Commission Directive 2006/63/EC, including dilution plating on semi-selective medium SMSA, indirect imunofluorescence (IIF, polymerase chain reaction (PCR using specific primers and bioassays on tomato plants, were strictly followed and the causal agent of the disease was identified as Ralstonia solanacearum. The identity of the pure cultures of the isolated organism was confirmed by PCR, IIF and pathogenicity tests on several other plant species (eggplant, tobacco, pelargonium and eucalyptus. In biovar determination, the failure of the isolates to utilise/oxidise certain carbon sources indicated that the isolates were all biovar 1. This biovar has a broader host range than biovar 2 strains, and affects several crops of economic importance including ornamental plants and forest trees. Comparative analysis of 16S rRNA and endoglucanase (egl gene sequences of these isolates with sequences that have been deposited at the GenBank revealed a similarity higher than 99% for several Ralstonia solanacearum isolates from biovar 1, including isolate DAR 64836 (Accession number DQ011551. This is the first report of Ralstonia solanacearum biovar 1 in Portugal. All control measures specified in the Commission Directive are being implemented.

  13. Ultrastructural aspects in perithecia hyphae septal pores of Glomerella cingulata F. SP. Phaseoli Ultraestrutura dos poros septais em hifas de peritécios de Glomerella cingulata f. sp. phaseoli

    OpenAIRE

    María Gabriela Roca M.; Maria das Graças Ongarelli; Lisete Chamma Davide; Maria Cristina Mendes-Costa

    2000-01-01

    Glomerella cingulata (Stonem.) Spauld. & Schrenk f. sp. phaseoli, better known in its anamorphic state Colletotrichum lindemuthianum (Sacc. & Magn.) Briosi & Cav., is a causal agent of anthracnose in beans (Phaseolus vulgaris L.). Ultrastructural aspects of the perithecial hyphae of this pathogen were studied. The perithecia hyphae septal pores were found either plugged by a vesicle or unplugged. Some perithecia hyphae septa presented no pore. The Woronin bodies, close to the septal pores, ap...

  14. Biovar Differentiation and Variation in Virulence of Ralstonia solanacearum Isolates Infecting Solanaceous Vegetables

    Directory of Open Access Journals (Sweden)

    Ram Devi Timila

    2016-12-01

    Full Text Available Bacterial wilt caused by Ralstonia solanacearum E.F. Smith is one of the destructive diseases of solanaceous vegetables specially tomato (Lycopersicon esculentum L. and eggplant (Solanum melongena L.. Experiments were conducted to determine biovar types existing among the strains or isolates of Nepal and variation in virulence in some vegetables belonging to solanaceae family. A total of 39 isolates infecting tomato, eggplant, chilli and potato collected from different parts of Nepal were analyzed for biovar types on the basis of 3 disaccharides and 3 hexose alcohols oxidation test. Experiments were conducted to determine variation in virulence or aggressiveness of some of the isolates under screen house conditions using three host differentials such as Pusa Ruby (susceptible, Bishesh (moderately resistant and Srijana (resistant tomato cultivars. Of the 39 isolates, 23 were biovar III, three biovar II, three biovar IV, and one was biovar I. Nine isolates could not be differentiated into any of the five biovars. For breeding and epidemiological purposes it is very important to analyze the variability of aggressiveness. A total of 5 isolates collected from different places were included in the test. Isolates from Bhaktapur was found the most virulent causing wilt in the variety Bishesh (moderately resistant. Other isolates had the negative impact with zero wilt on the differentials used. Isolates from Jungekhola of Dhading district did not induce wilt even on susceptible variety (Pusa Ruby, but exhibited only senescence reaction. The result indicated that there is some slight variation among the isolates tested. Some effective management tactics might be needed in those locations where highly aggressive or virulent strain of bacterial wilt is prevalent, because resistant variety may not be stable in such locations.

  15. Reaction of common bean varieties ( Phaseolus vulgaris L. to the infection for Uromyces phaseoli (Pers. Wint var. typica Arth

    Directory of Open Access Journals (Sweden)

    Arth Cabrera

    2014-01-01

    Full Text Available It was evaluated the reaction of twenty-five common bean varieties to Uromyces phaseoli (Pers.Wintvar. typica Arth infection using the methodology informed by the Colombian International Center of Tropical Agriculture (CIAT. These experiments were developed in the Agricultural Experimental Station belonging to the Agricultural Sciences Faculty of Las Villas Central University, under a design of random blocks with four repetitions in the period between January to April 2011. The evaluations of infection intensity and pustule type were carried out weekly starting from the beginning of disease symptoms in 15 plants for varieties. Results evidenced differences in the response of varieties as well as a different behavior to the infection for this phytopathogen fungus. Guama 23 variety was classified as resistant highly to the infection for U. phaseoli. This work constitutes a fundamental tool for future programs on genetic improvement of common bean cultivation in Cuba.

  16. Cloning of a two-component signal transduction system of Xanthomonas campestris pv. phaseoli var. fuscans strain BXPF65

    DEFF Research Database (Denmark)

    Chan, JWYF; Maynard, Scott; Goodwin, PH

    1998-01-01

    A putative two-component signal transduction system was amplified and cloned from the plant pathogenic bacterium Xanthomonas campestris pv. phaseoli var. fuscans isolate BXPF65. The 620 bp amplified fragment was sequenced and analyzed with the BLAST Enhanced Alignment Utility (BEAUTY). BEAUTY...... that the putative histidine kinase has homology with conserved “transmitter” domains of sensor proteins in two-component signal transduction systems. RFLP analysis using the putative signal transduction system showed polymorphisms among the strains....

  17. [Algorithm of toxigenic genetically altered Vibrio cholerae El Tor biovar strain identification].

    Science.gov (United States)

    Smirnova, N I; Agafonov, D A; Zadnova, S P; Cherkasov, A V; Kutyrev, V V

    2014-01-01

    Development of an algorithm of genetically altered Vibrio cholerae biovar El Tor strai identification that ensures determination of serogroup, serovar and biovar of the studied isolate based on pheno- and genotypic properties, detection of genetically altered cholera El Tor causative agents, their differentiation by epidemic potential as well as evaluation of variability of key pathogenicity genes. Complex analysis of 28 natural V. cholerae strains was carried out by using traditional microbiological methods, PCR and fragmentary sequencing. An algorithm of toxigenic genetically altered V. cholerae biovar El Tor strain identification was developed that includes 4 stages: determination of serogroup, serovar and biovar based on phenotypic properties, confirmation of serogroup and biovar based on molecular-genetic properties determination of strains as genetically altered, differentiation of genetically altered strains by their epidemic potential and detection of ctxB and tcpA key pathogenicity gene polymorphism. The algorithm is based on the use of traditional microbiological methods, PCR and sequencing of gene fragments. The use of the developed algorithm will increase the effectiveness of detection of genetically altered variants of the cholera El Tor causative agent, their differentiation by epidemic potential and will ensure establishment of polymorphism of genes that code key pathogenicity factors for determination of origins of the strains and possible routes of introduction of the infection.

  18. Genome sequence of Corynebacterium pseudotuberculosis biovar equi strain 258 and prediction of antigenic targets to improve biotechnological vaccine production

    DEFF Research Database (Denmark)

    Soares, Siomar C; Trost, Eva; Ramos, Rommel T J

    2013-01-01

    Corynebacterium pseudotuberculosis is the causative agent of several veterinary diseases in a broad range of economically important hosts, which can vary from caseous lymphadenitis in sheep and goats (biovar ovis) to ulcerative lymphangitis in cattle and horses (biovar equi). Existing vaccines ag...

  19. JST Thesaurus Headwords and Synonyms: Brucella melitensis biovar abortus [MeCab user dictionary for science technology term[Archive

    Lifescience Database Archive (English)

    Full Text Available MeCab user dictionary for science technology term Brucella melitensis biovar abortu...s 名詞 一般 * * * * ウシ流産菌 ウシリュウザンキン ウシリューザンキン Thesaurus2015 200906011165481664 C LS07 UNKNOWN_2 Brucella melitensis biovar abortus

  20. [Effect of indolylacetic acid on formation of bacteroid forms of Rhizobium leguminosarum].

    Science.gov (United States)

    Lobanok, E V; Bakanchikova, T I

    1979-01-01

    The purpose of this work was to study the effect of indolylacetic acid (IAA) on the strains of Rhizobium leguminosarum, effective and noneffective with respect to symbiotic nitrogen fixation (L4 and 245a, and 14--73, respectively). IAA at a concentration of 50 mcg/ml and higher inhibited the growth of the bacterium, temporarily delayed celular division, and induced intensive formation of elongated bacteroid-like cells, predominantly Y-shaped or having a clavate shape. Many bacteroid-like cells were capable of division after a certain delay.

  1. Selectable genes for transformation of the fungal plant pathogen Glomerella cingulata f. sp. phaseoli (Colletotrichum lindemuthianum).

    Science.gov (United States)

    Rodriguez, R J; Yoder, O C

    1987-01-01

    Glomerella cingulata f. sp. phaseoli (Gcp) was transformed using either of two selectable markers: the amdS + gene of Aspergillus nidulans, which encodes acetamidase and permits growth on acetamide as the sole nitrogen source and the hygBR gene of Escherichia coli which encodes hygromycin B (Hy) phosphotransferase and permits growth in the presence of the antibiotic Hy. The amdS+ gene functioned in Gcp under control of A. nidulans regulatory signals and hygBR was expressed after fusion to a promoter from Cochliobolus heterostrophus, another filamentous ascomycete. Protoplasts to be transformed were generated with the digestive enzyme complex Novozym 234 and then were exposed to plasmid DNA in the presence of 10 mM CaCl2 and polyethylene glycol. Transformation occurred by integration of single or multiple copies of either the amdS+ or hygBR plasmid into the fungal genome. There was no evidence of autonomous plasmid replication. Transformants were mitotically stable on selective and nonselective media. However, transforming DNA in hygBR transformants was observed to occasionally rearrange during nonselective growth, resulting in fewer copies of the plasmid per genome. These transformants were capable of infecting bean (Phaseolus vulgaris), the Gcp host plant, and after recovery from infected tissue were found to have retained both the transforming DNA unrearranged in their genomes and the Hy resistance phenotype. All single-conidial cultures derived from both amdS+ and hygBR transformants had the transplanted phenotype, suggesting that transformants were homokaryons.

  2. Reação de linhagens de feijoeiro ao fungo Fusarium oxysporum f. sp. phaseoli em condições controladas Reaction of common bean lines to Fusarium oxysporum f. sp. phaseoli in controlled conditions

    Directory of Open Access Journals (Sweden)

    Mônica Juliani Zavaglia Pereira

    2011-10-01

    Full Text Available Entre os patógenos que mais contribuem para a redução da produtividade do feijoeiro no Brasil está o Fusarium oxysporum f. sp. phaseoli. O emprego de cultivares resistentes é o controle mais eficaz para esse patógeno. Conduziu-se este trabalho, com o objetivo de avaliar linhagens de feijoeiro (Phaseolus vulgaris L. do banco de germoplasma da Universidade Federal de Lavras (UFLA, quanto a reação ao Fusarium oxysporum f. sp. phaseoli e, ao mesmo tempo, estimar os parâmetros genéticos e fenotípicos que possam auxiliar em futuros programas de melhoramento para esse caráter. Foram avaliadas 367 linhagens em dez experimentos. As testemunhas 'Carioca' (suscetível e 'Carioca MG' (resistente foram utilizadas em todos os experimentos. O delineamento utilizado foi inteiramente ao acaso, com cinco repetições e parcelas de uma planta por vaso. As inoculações foram realizadas segundo a metodologia de corte e imersão de raízes na suspensão de esporos do fungo e as avaliações realizadas aos 21 dias após a inoculação com base no índice de severidade da doença empregando-se notas de 1 (plantas sem sintomas a 9 (plantas mortas.Das linhagens do banco de germoplasma da Universidade Federal de Lavras (UFLA avaliadas, 36,5% foram resistentes ao Fusarium oxysporum f. sp. phaseoli. Entre as resistentes, a maioria foi linhagens obtidas antes de 1990. Das 18 linhagens dos experimentos de VCU, do período de 2005/06, apenas quatro foram suscetíveis. A estimativa da herdabilidade (h² foi elevada (h² = 87%, indicando que, a princípio, o caráter é de fácil seleção.Among the pathogens that most contribute for reducing the productivity of beans in Brazil is the Fusarium oxysporum f. sp. phaseoli. The use of resistant cultivars is the most effective control for this pathogen. The aim of this study was to evaluate germoplasm bean (Phaseolus vulgaris L. lines of the Federal University of Lavras (UFLA as the reaction to the Fusarium oxysporum f

  3. Genetic Diversity among Rhizobium leguminosarum bv. Trifolii Strains Revealed by Allozyme and Restriction Fragment Length Polymorphism Analyses

    Science.gov (United States)

    Demezas, David H.; Reardon, Terry B.; Watson, John M.; Gibson, Alan H.

    1991-01-01

    Allozyme electrophoresis and restriction fragment length polymorphism (RFLP) analyses were used to examine the genetic diversity of a collection of 18 Rhizobium leguminosarum bv. trifolii, 1 R. leguminosarum bv. viciae, and 2 R. meliloti strains. Allozyme analysis at 28 loci revealed 16 electrophoretic types. The mean genetic distance between electrophoretic types of R. leguminosarum and R. meliloti was 0.83. Within R. leguminosarum, the single strain of bv. viciae differed at an average of 0.65 from strains of bv. trifolii, while electrophoretic types of bv. trifolii differed at a range of 0.23 to 0.62. Analysis of RFLPs around two chromosomal DNA probes also delineated 16 unique RFLP patterns and yielded genetic diversity similar to that revealed by the allozyme data. Analysis of RFLPs around three Sym (symbiotic) plasmid-derived probes demonstrated that the Sym plasmids reflect genetic divergence similar to that of their bacterial hosts. The large genetic distances between many strains precluded reliable estimates of their genetic relationships. PMID:16348600

  4. Bacterial leaf rot of Aloe vera L., caused byErwinia chrysanthemi biovar 3

    NARCIS (Netherlands)

    Laat, de P.C.A.; Verhoeven, J.T.W.; Danse, J.D.

    1994-01-01

    A severe attack of the bacteriumErwinia chrysantemi biovar 3 on the succulentAloe vera on the Carribean island of Aruba is described. Biochemical and pathological characteristics of strains are presented, including results of successful inoculation experiments onAloe vera. This is the first report

  5. Pig slurry reduces the survival of Raltstonia solanacearum biovar 2 in soil

    NARCIS (Netherlands)

    Gorissen, A.; Overbeek, van L.S.; Elsas, van J.D.

    2004-01-01

    The effect of added pig slurry and solarization on the survival of Ralstonia solanacearum biovar 2 strain 1609 in soil was analysed in soil microcosms and field plots. In addition, the invasion of potato plants by R. solanacearum and the development of disease symptoms were determined, as measures

  6. Differences in outer membrane proteins of the lymphogranuloma venereum and trachoma biovars of Chlamydia trachomatis

    International Nuclear Information System (INIS)

    Batteiger, B.E.; Jones, R.B.

    1985-01-01

    The lymphogranuloma venereum (LGV) and trachoma biovars of Chlamydia trachomatis exhibit differences in biological properties both in vivo and in vitro. To identify analogous biochemical differences, the authors studied the molecular charges of chlamydial outer membrane proteins (OMPs) by means of isoelectric focusing and nonequilibrium pH gradient electrophoresis. Analysis of proteins of whole elementary bodies biosynthetically labeled with L-[35S]cysteine revealed that most chlamydial proteins were neutral or acidic. The major OMPs (MOMPs) of all strains tested were acidic and had apparent isoelectric points (pIs) that varied within narrow limits despite differences in molecular mass of up to 3,000 daltons (Da). However, a low-molecular-mass cysteine-rich OMP analogous to that previously described for Chlamydia psittaci varied consistently in molecular mass (12,500 versus 12,000 Da) and pI (5.4 versus 6.9) between LGV strains and trachoma strains, respectively. OMPs with a molecular mass of 60,000 Da in the trachoma biovar strains had pIs in the 7.3 to 7.7 range. However, analogous OMPs in the LGV strains existed as a doublet with a molecular mass of about 60,000 Da. These data indicate substantial differences in biochemical characteristics of analogous OMPs in the LGV and trachoma biovars. Such differences are the first structural differences described between LGV and trachoma strains which support their distinction into separate biovars and may be related to some of their biological differences

  7. Genotipos de frijol (Phaseolus Vulgaris l. resistentes a Xanthomonas campestris pv. phaseoli de Mexico

    Directory of Open Access Journals (Sweden)

    Rosa Navarrete

    2000-01-01

    Full Text Available Genotipos de frijol (Phaseolus vulgaris L. resistentes a Xanthomonas campestris pv. phaseoli de México. Durante 1995 se evaluó la reacción de genotipos de frijol de diversos origenes a Xcp, bajo condiciones de invernadero en el Campo Experimental del Valle de México, del INIFAP. Se realizaron tres experimentos con a120, b44 y csiete genotipos de frijol. Las plantas se inocularon por corte con navajas en la etapa V3, a y b con una mezcla de nueve cepas de Xcp y el c, con cada una de siete cepas con diferente grado de patogenicidad. La severidad se evaluó 20 días después de la inoculación, por comparación con una escala visual de nueve grados. Los datos se analizaron bajo un diseño completamente al azar. En a, los genotipos que mostraron reacción de resistencia a Xcp fueron: A 36, A 475, G 5686, G 11867, Harowood, SEA 14, XAN 266, MCD 4012 y REN 27. En b los genotipos resistentes fueron: Sequía Durango, Taylor y XAN 30. En los experimentos anteriores la severidad de la enfermedad mostró una distribución normal, con el máximo número de genotipos en el grado de severidad cinco en a y seis en b. Los resultados obtenidos indican que el uso de mezclas de cepas de bacterias con diferente patogenicidad es eficiente para identificar genotipos de frijol resistentes a Xcp. Los genotipos resistentes identificados en el último experimento, mostraron respuesta diferencial e interacciones genotipo por cepa. REN 27 y SEA 14 mostraron resistencia a las cepas utilizadas

  8. Assessment of Qualitative and Quantitative Characters of Two Persian Clover Ecotypes Inoculated by Rhizobium leguminosarum biovartrifoli and Pesudomonas putida Bacteria

    Directory of Open Access Journals (Sweden)

    R Azamei

    2016-02-01

    Full Text Available Introduction Over the past decades, world attitude has changed towards the reduction of environmental pollutants. Harmful effects of synthetic fertilizers on environment have been identified. Bio-fertilizers are not harmful to the environment, but also they have favorable effects on plant growth processes. Soil biotechnology can be defined as the study of soil organisms and their metabolic processes which may have positive effects on plant yields. The main goal of this study is to asess the biotechnology fertilizers beneficial effects on soil organisms and their subsequently to maximize the yield. It is also our desire conside the soil quality, hygiene and environmental protection along this process. Among the strain of nitrogen-fixing bacteria, symbiotic bacteria such as rhizobium bacteria are important and essential in planning the sustainable farming systems. Several studies have shown that crop varieties which inoculated with rhizobium and pseudomonas were superior in yield production and performance. Material and Methods An experiment was designed as factorial performed in randomized complete block design (RCBD with three replications in Agricultural Research Center of Golpayegan (Isfahan during 2010 – 2011. the purpose of this study was to evaluate the effects of inoculation of two ecotypes of Persian clover by various strains of Rhizobium leguminosarum. Biovar trifoli bacteria accompanied with plant growth promoting rhizobacteria (PGPR Pseudomonas putida was employed to find certain qualitative and quantitative characteristics of clover yield, The main plots included two local ecotypes of Persian clover; Arak Haft Chin (V1 and Isfahan Haft Chin (V2, the subplots included inoculation by two strain of Rhizobium; Rb-3, Rb-13 and one strain of Pseudomonas; PS -168.4 cuts were performed during the experiment and 60 kg/ha seed was used for cultivation based on local knowledge. According to recommendations of the Institute of Soil and Water

  9. Evolution and genome specialization of Brucella suis biovar 2 Iberian lineages.

    Science.gov (United States)

    Ferreira, Ana Cristina; Tenreiro, Rogério; de Sá, Maria Inácia Corrêa; Dias, Ricardo

    2017-09-12

    Swine brucellosis caused by B. suis biovar 2 is an emergent disease in domestic pigs in Europe. The emergence of this pathogen has been linked to the increase of extensive pig farms and the high density of infected wild boars (Sus scrofa). In Portugal and Spain, the majority of strains share specific molecular characteristics, which allowed establishing an Iberian clonal lineage. However, several strains isolated from wild boars in the North-East region of Spain are similar to strains isolated in different Central European countries. Comparative analysis of five newly fully sequenced B. suis biovar 2 strains belonging to the main circulating clones in Iberian Peninsula, with publicly available Brucella spp. genomes, revealed that strains from Iberian clonal lineage share 74% similarity with those reference genomes. Besides the 210 kb translocation event present in all biovar 2 strains, an inversion with 944 kb was presented in chromosome I of strains from the Iberian clone. At left and right crossover points, the inversion disrupted a TRAP dicarboxylate transporter, DctM subunit, and an integral membrane protein TerC. The gene dctM is well conserved in Brucella spp. except in strains from the Iberian clonal lineage. Intraspecies comparative analysis also exposed a number of biovar-, haplotype- and strain-specific insertion-deletion (INDELs) events and single nucleotide polymorphisms (SNPs) that could explain differences in virulence and host specificities. Most discriminative mutations were associated to membrane related molecules (29%) and enzymes involved in catabolism processes (20%). Molecular identification of both B. suis biovar 2 clonal lineages could be easily achieved using the target-PCR procedures established in this work for the evaluated INDELs. Whole-genome analyses supports that the B. suis biovar 2 Iberian clonal lineage evolved from the Central-European lineage and suggests that the genomic specialization of this pathogen in the Iberian Peninsula

  10. Emended description of Campylobacter sputorum and revision of its infrasubspecific (biovar) divisions, including C-sputorum biovar paraureolyticus, a urease-producing variant from cattle and humans

    DEFF Research Database (Denmark)

    On, S.L.W.; Atabay, H.I.; Corry, J.E.L.

    1998-01-01

    A polyphasic taxonomic study of 15 bovine and human strains assigned to the catalase-negative, urease-positive campylobacter (CNUPC) group identified these bacteria as a novel, ureolytic biovar of Campylobacter sputorum for which we propose the name C. sputorum bv. paraureolyticus: suitable...... should be revised to include by. sputorum for catalase-negative strains; by. fecalis for catalase-positive strains; and by. paraureolyticus for urease-positive strains. Strains classified previously as by. bubulus should be reclassified as by. sputorum. The species description of C. sputorum is revised...

  11. Duplex PCR for differentiation of the vaccine strain Brucella suis S2 and B. suis biovar 1 from other strains of Brucella spp.

    Science.gov (United States)

    Nan, Wenlong; Tan, Pengfei; Wang, Yong; Xu, Zouliang; Mao, Kairong; Peng, Daxin; Chen, Yiping

    2014-09-01

    Immunisation with attenuated Brucella spp. vaccines prevents brucellosis, but may also interfere with diagnosis. In this study, a duplex PCR was developed to distinguish Brucella suis vaccine strain S2 from field strains of B. suis biovar 1 and other Brucella spp. The PCR detected 60 fg genomic DNA of B. suis S2 or biovar 1 field strains and was able to distinguish B. suis S2 and wild-type strains of B. suis biovar 1 among 76 field isolates representing all the common species and biovars, as well as four vaccine strains, of Brucella. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. Detection of Xanthomonas axonopodis pv. Phaseoli and Pseudomonas savastanoi pv. Phaseolicola on bean seed using a Milk-tween medium

    Directory of Open Access Journals (Sweden)

    Popović Tatjana

    2012-01-01

    Full Text Available Bean production is threaten by phytopathogenic bacteria causing agents of blights, Xanthomonas axonopodis pv. phaseoli (Xap and Pseudomonas savastanoi pv. phaseolicola (Psp. Since there is no satisfactory chemical control for the disease, the recommended measures are preventive and include use of healthy seed, crop rotation, deep plowing and use of resistant cultivars. In this work we involved a detection method for isolation of Xap and Psp from bean seed to semi-selective medium Milk Agar Tween (MT. On this medium, Xap formed yellow, mucoid and convex colonies with two hydrolysis zones (less milk and more enlightened, and Psp formed whitish-cream, flat and round colonies. The identification of Xap and Psp was confirmed using the ELISA and PCR. Due to its selectivity, easy preparation and possibility of simultaneous detection of bacteria Xap and Psp, MT medium can be recommended for routine test of seed health for local seed or seed from import.

  13. Rhizobium leguminosarum COMO ORGANISMO BIOCONTROLADOR DE LA INTERACCIÓN HOSPEDERO-PATÓGENO: CLAVEL (Dianthus caryophyllus – Fusarium Oxysporum f. sp. dianthi

    Directory of Open Access Journals (Sweden)

    Cheol Woo Lee Park

    2010-08-01

    Full Text Available Se investigó el efecto de biocontrol de Rhizobium leguminosarum (R. leguminosarum cepa B, contra Fusarium oxysporum f.sp.dianthi (FOD raza 2, en la interacción Clavel - FOD. Se utilizó la raza 2 de FOD por ser la de mayor patogenicidad y distribución en las fincas de cultivo de clavel en Colombia. Para ello se establecieron las condiciones de inoculación de FOD sobre cultivos establecidos de R . leguminosarum, variando la concentración in vitro de la bacteria. Se encontró una reducción en el número de microconidias hasta un 90% y una inhibición en el crecimiento radial del patógeno de hasta un 71 %. En el ensayo de microcultivo dual se detectó fraccionamiento de las hifas después de 48 horas de incubación con R . leguminosarum En el ensayo in vivo, los esquejes de clavel Raggio di solé, variedad susceptible al patógeno, fueron inoculados con 45.0 x 10 células de R. leguminosarum por matero, mostrando una severidad inferior al 5%, una incidencia menor del 20% y una reducción del índice de la enfermedad hasta de un 92% en presencia del patógeno.

  14. Typing Discrepancy Between Phenotypic and Molecular Characterization Revealing an Emerging Biovar 9 Variant of Smooth Phage-Resistant B. abortus Strain 8416 in China.

    Science.gov (United States)

    Kang, Yao-Xia; Li, Xu-Ming; Piao, Dong-Ri; Tian, Guo-Zhong; Jiang, Hai; Jia, En-Hou; Lin, Liang; Cui, Bu-Yun; Chang, Yung-Fu; Guo, Xiao-Kui; Zhu, Yong-Zhang

    2015-01-01

    A newly isolated smooth colony morphology phage-resistant strain 8416 isolated from a 45-year-old cattle farm cleaner with clinical features of brucellosis in China was reported. The most unusual phenotype was its resistance to two Brucella phages Tbilisi and Weybridge, but sensitive to Berkeley 2, a pattern similar to that of Brucella melitensis biovar 1. VITEK 2 biochemical identification system found that both strain 8416 and B. melitensis strains shared positive ILATk, but negative in other B. abortus strains. However, routine biochemical and phenotypic characteristics of strain 8416 were most similar to that of B. abortus biovar 9 except CO2 requirement. In addition, multiple PCR molecular typing assays including AMOS-PCR, B. abortus special PCR (B-ab PCR) and a novel sub-biovar typing PCR, indicated that strain 8416 may belong to either biovar 3b or 9 of B. abortus. Surprisingly, further MLVA typing results showed that strain 8416 was most closely related to B. abortus biovar 3 in the Brucella MLVA database, primarily differing in 4 out of 16 screened loci. Therefore, due to the unusual discrepancy between phenotypic (biochemical reactions and particular phage lysis profile) and molecular typing characteristics, strain 8416 could not be exactly classified to any of the existing B. abortus biovars and might be a new variant of B. abortus biovar 9. The present study also indicates that the present phage typing scheme for Brucella sp. is subject to variation and the routine Brucella biovar typing needs further studies.

  15. Genome sequencing reveals a new lineage associated with lablab bean and genetic exchange between Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans

    Directory of Open Access Journals (Sweden)

    Valente eAritua

    2015-10-01

    Full Text Available Common bacterial blight is a devastating seed-borne disease of common beans that also occurs on other legume species including lablab and Lima beans. We sequenced and analysed the genomes of 26 isolates of Xanthomonas axonopodis pv. phaseoli and X. fuscans subsp. fuscans, the causative agents of this disease, collected over four decades and six continents. This revealed considerable genetic variation within both taxa, encompassing both single-nucleotide variants and differences in gene content, that could be exploited for tracking pathogen spread. The bacterial isolate from Lima bean fell within the previously described Genetic Lineage 1, along with the pathovar type isolate (NCPPB 3035. The isolates from lablab represent a new, previously unknown genetic lineage closely related to strains of X. axonopodis pv. glycines. Finally, we identified more than 100 genes that appear to have been recently acquired by Xanthomonas axonopodis pv. phaseoli from X. fuscans subsp. fuscans.

  16. Transformation of pWWO in Rhizobium leguminosarum DPT to Engineer Toluene Degrading Ability for Rhizoremediation.

    Science.gov (United States)

    Goel, Garima; Pandey, Piyush; Sood, Anchal; Bisht, Sandeep; Maheshwari, D K; Sharma, G D

    2012-06-01

    Rhizoremediation of organic xenobiotics is based on interactions between plants and their associated micro-organisms. The present work was designed to engineer a bacterial system having toluene degradation ability along with plant growth promoting characteristics for effective rhizoremediation. pWWO harboring the genes responsible for toluene breakdown was isolated from Pseudomonas putida MTCC 979 and successfully transformed in Rhizobium DPT. This resulted in a bacterial strain (DPT(T)) which had the ability to degrade toluene as well as enhance growth of host plant. The frequency of transformation was recorded 5.7 × 10(-6). DPT produced IAA, siderophore, chitinase, HCN, ACC deaminase, solubilized inorganic phosphate, fixed atmospheric nitrogen and inhibited the growth of Fusarium oxysporum and Macrophomina phaseolina in vitro. During pot assay, 50 ppm toluene in soil was found to inhibit the germination of Cajanus cajan seeds. However when the seeds bacterized with toluene degrading P. putida or R. leguminosarum DPT were sown in pots, again no germination was observed. Non-bacterized as well as bacterized seeds germinated successfully in toluene free soil as control. The results forced for an alternative mode of application of bacteria for rhizoremediation purpose. Hence bacterial suspension was mixed with soil having 50 ppm of toluene. Germination index in DPT treated soil was 100% while in P. putida it was 50%. Untreated soil with toluene restricted the seeds to germinate.

  17. Rhizobium leguminosarum bv. trifolii rosR is required for interaction with clover, biofilm formation and adaptation to the environment

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    Piersiak Tomasz

    2010-11-01

    Full Text Available Abstract Background Rhizobium leguminosarum bv. trifolii is a symbiotic nitrogen-fixing bacterium that elicits nodules on roots of host plants Trifolium spp. Bacterial surface polysaccharides are crucial for establishment of a successful symbiosis with legumes that form indeterminate-type nodules, such as Trifolium, Pisum, Vicia, and Medicago spp. and aid the bacterium in withstanding osmotic and other environmental stresses. Recently, the R. leguminosarum bv. trifolii RosR regulatory protein which controls exopolysaccharide production has been identified and characterized. Results In this work, we extend our earlier studies to the characterization of rosR mutants which exhibit pleiotropic phenotypes. The mutants produce three times less exopolysaccharide than the wild type, and the low-molecular-weight fraction in that polymer is greatly reduced. Mutation in rosR also results in quantitative alterations in the polysaccharide constituent of lipopolysaccharide. The rosR mutants are more sensitive to surface-active detergents, antibiotics of the beta-lactam group and some osmolytes, indicating changes in the bacterial membranes. In addition, the rosR mutants exhibit significant decrease in motility and form a biofilm on plastic surfaces, which differs significantly in depth, architecture, and bacterial viability from that of the wild type. The most striking effect of rosR mutation is the considerably decreased attachment and colonization of root hairs, indicating that the mutation affects the first stage of the invasion process. Infection threads initiate at a drastically reduced rate and frequently abort before they reach the base of root hairs. Although these mutants form nodules on clover, they are unable to fix nitrogen and are outcompeted by the wild type in mixed inoculations, demonstrating that functional rosR is important for competitive nodulation. Conclusions This report demonstrates the significant role RosR regulatory protein plays in

  18. Role of O2 in the Growth of Rhizobium leguminosarum bv. viciae 3841 on Glucose and Succinate

    OpenAIRE

    Wheatley, Rachel M.; Ramachandran, Vinoy K.; Geddes, Barney A.; Perry, Benjamin J.; Yost, Chris K.; Poole, Philip S.

    2016-01-01

    Insertion sequencing (INSeq) analysis of Rhizobium leguminosarum bv. viciae 3841 (Rlv3841) grown on glucose or succinate at both 21% and 1% O2 was used to understand how O2 concentration alters metabolism. Two transcriptional regulators were required for growth on glucose (pRL120207 [eryD] and RL0547 [phoB]), five were required on succinate (pRL100388, RL1641, RL1642, RL3427, and RL4524 [ecfL]), and three were required on 1% O2 (pRL110072, RL0545 [phoU], and RL4042). A novel toxin-antitoxin s...

  19. Bacterial wilt of potato (Ralstonia solanacearum race 3, biovar 2): disease management, pathogen survival and possible eradication

    NARCIS (Netherlands)

    Messiha, N.A.S.

    2006-01-01

    Potato brown rot, caused by Ralstonia solanacearum race 3 biovar 2 (Phylovar II, sequevar 1), is a serious endemic disease in the Nile Delta of Egypt. It is a quarantine disease in the EU, and export of potatoes from

  20. Competitive nodulation blocking of cv. Afghanistan pea is related to high levels of nodulation factors made by some strains of Rhizobium leguminosarum bv. viciae

    NARCIS (Netherlands)

    Hogg, B.; Davies, A.E.; Wilson, K.E.; Bisseling, T.; Downie, J.A.

    2002-01-01

    Cultivar Afghanistan peas are resistant to nodulation by many strains of Rhizobium leguminosarum bv. viciae but are nodulated by strain TOM, which carries the host specificity gene nodX. Some strains that lack nodX can inhibit nodulation of cv. Afghanistan by strain TOM. We present evidence that

  1. Plasmid Negative Regulation of CPAF Expression Is Pgp4 Independent and Restricted to Invasive Chlamydia trachomatis Biovars

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    Michael John Patton

    2018-01-01

    Full Text Available Chlamydia trachomatis is an obligate intracellular bacterial pathogen that causes blinding trachoma and sexually transmitted disease. C. trachomatis isolates are classified into 2 biovars—lymphogranuloma venereum (LGV and trachoma—which are distinguished biologically by their natural host cell infection tropism. LGV biovars infect macrophages and are invasive, whereas trachoma biovars infect oculo-urogenital epithelial cells and are noninvasive. The C. trachomatis plasmid is an important virulence factor in the pathogenesis of these infections. Central to its pathogenic role is the transcriptional regulatory function of the plasmid protein Pgp4, which regulates the expression of plasmid and chromosomal virulence genes. As many gene regulatory functions are post-transcriptional, we employed a comparative proteomic study of cells infected with plasmid-cured C. trachomatis serovars A and D (trachoma biovar, a L2 serovar (LGV biovar, and the L2 serovar transformed with a plasmid containing a nonsense mutation in pgp4 to more completely elucidate the effects of the plasmid on chlamydial infection biology. Our results show that the Pgp4-dependent elevations in the levels of Pgp3 and a conserved core set of chromosomally encoded proteins are remarkably similar for serovars within both C. trachomatis biovars. Conversely, we found a plasmid-dependent, Pgp4-independent, negative regulation in the expression of the chlamydial protease-like activity factor (CPAF for the L2 serovar but not the A and D serovars. The molecular mechanism of plasmid-dependent negative regulation of CPAF expression in the LGV serovar is not understood but is likely important to understanding its macrophage infection tropism and invasive infection nature.

  2. Typing discrepancy between phenotypic and molecular characterization revealing an emerging biovar 9 variant of smooth phage-resistant B. abortus strain 8416 in China

    Directory of Open Access Journals (Sweden)

    YaoXia eKang

    2015-12-01

    Full Text Available A newly isolated smooth colony morphology phage-resistant (SPR strain 8416 isolated from a 45-year-old cattle farm cleaner with clinical features of brucellosis in China was reported. The most unusual phenotype was its resistance to two Brucella phages Tbilisi and Weybridge, but sensitive to Berkeley 2, a pattern similar to that of B. melitensis biovar 1. VITEK 2 biochemical identification system found that both strain 8416 and B. melitensis strains shared positive ILATk, but negative in other B. abortus strains. However, routine biochemical and phenotypic characteristics of strain 8416 were most similar to that of B. abortus biovar 9 except CO2 requirement. In addition, multiple PCR molecular typing assays including AMOS-PCR, B. abortus special PCR (B-ab PCR and a novel sub-biovar typing PCR, indicated that strain 8416 may belong to either biovar 3b or 9 of B. abortus. Surprisingly, further MLVA typing results showed that strain 8416 was most closely related to B. abortus biovar 3 in the Brucella MLVA database, primarily differing in 4 out of 16 screened loci. Therefore, due to the unusual discrepancy between phenotypic (biochemical reactions and particular phage lysis profile and molecular typing characteristics, strain 8416 couldn’t be exactly classified to any of the existing B. abortus biovars and might be a new variant of B. abortus biovar 9. The present study also indicates that the present phage typing scheme for Brucella spp. is subject to variation and the routine Brucella biovar typing needs further studies.

  3. Phenotypic and genotypic characterization of Brucella strains isolated from autochthonous livestock reveals the dominance of B. abortus biovar 3a in Nigeria.

    Science.gov (United States)

    Bertu, Wilson J; Ducrotoy, Marie J; Muñoz, Pilar M; Mick, Virginie; Zúñiga-Ripa, Amaia; Bryssinckx, Ward; Kwaga, Jacob K P; Kabir, Junaid; Welburn, Susan C; Moriyón, Ignacio; Ocholi, Reuben A

    2015-10-22

    Brucellosis is a worldwide widespread zoonosis caused by bacteria of the genus Brucella. Control of this disease in a given area requires an understanding of the Brucella species circulating in livestock and humans. However, because of the difficulties intrinsic to Brucella isolation and typing, such data are scarce for resource-poor areas. The paucity of bacteriological data and the consequent imperfect epidemiological picture are particularly critical for Sahelian and Sub-Sahara African countries. Here, we report on the characterization of 34 isolates collected between 1976 and 2012 from cattle, sheep and horses in Nigeria. All isolates were identified as Brucella abortus by Bruce-ladder PCR and assigned to biovar 3 by conventional typing. Further analysis by enhanced AMOS-ERY PCR showed that all of them belonged to the 3a sub-biovar, and MLVA analysis grouped them in a cluster clearly distinct from that formed by European B. abortus biovar 3b strains. Nevertheless, MLVA detected heterogeneity within the Nigerian biovar 3a strains. The close genetic profiles of the isolates from cattle, sheep and horses, suggest that, at least in some parts of Nigeria, biovar 3a circulates among animal species that are not the preferential hosts of B. abortus. Consistent with previous genetic analyses of 7 strains from Ivory Cost, Gambia and Togo, the analysis of these 34 Nigerian strains supports the hypothesis that the B. abortus biovar 3a lineage is dominant in West African countries. Copyright © 2015 Elsevier B.V. All rights reserved.

  4. Colonização radicular de plantas cultivadas por Ralstonia solanacearum biovares 1, 2 e 3 Root colonization of cultivated plants inoculated with Ralstonia solanacearum biovar 1, 2 and 3

    Directory of Open Access Journals (Sweden)

    José Magno Martins Bringel

    2001-09-01

    Full Text Available A murcha bacteriana causada por Ralstonia solanacearum é considerada a principal doença de origem bacteriana no mundo. Centenas de espécies de plantas pertencentes a mais de 50 famílias botânicas têm sido relatadas como hospedeiras. Foi avaliada, em condições de casa de vegetação, a colonização radicular de alface, arroz, cebolinha, ervilha, pepino e soja, por seis isolados de Ralstonia solanacearum (Rs, biovares 1, 2 e 3. Estirpes dos isolados de Rs com resistência múltipla aos antibióticos estreptomicina, rifampicina e cloranfenicol foram utilizadas. A colonização foi avaliada 45 dias após a inoculação, através do plaqueamento de suspensão de trituração em meio de cultura semi-seletivo. A ervilha comportou-se como hospedeira de todos os isolados mas, apenas um isolado da biovar 3 foi patogênico a esta espécie. A soja apresentou populações elevadas de quatro isolados distribuídos entre as três biovares e o pepino, de apenas dois isolados das biovares 1 e 3. Exceto para o isolado que foi patogênico à ervilha, as plantas não apresentaram sintomas da doença, comportando-se como hospedeiras não suscetível. O arroz apresentou populações muito baixas de todos os isolados. Alface e cebolinha não hospedaram nenhum dos isolados inoculados. Os resultados mostram a capacidade de Rs colonizar e sobreviver em diferentes espécies de plantas como rizobactérias.Bacterial wilt caused by Ralstonia solanacearum is considered the main plant disease of bacterial origin in the world, where hundreds of plant species in more than 50 botanical families are host plants. Root colonization of lettuce (Lactuca sativa, rice (Oryza sativa, spring onion (Allium fistulosum, pea (Pisum sativum, cucumber (Cucumis sativus, and soybean (Glycine max by six isolates of Ralstonia solanacearum (Rs biovars 1, 2 and 3 was evaluated under greenhouse conditions. Bacterial strains resistant to streptomycin, rifampicin and chloranfenicol were used

  5. A Rhizobium leguminosarum CHDL- (Cadherin-Like-) Lectin Participates in Assembly and Remodeling of the Biofilm Matrix

    DEFF Research Database (Denmark)

    Vozza, Nicolás F.; Abdian, Patricia L; Russo, Daniela M

    2016-01-01

    In natural environments most bacteria live in multicellular structures called biofilms. These cell aggregates are enclosed in a self-produced polymeric extracellular matrix, which protects the cells, provides mechanical stability and mediates cellular cohesion and adhesion to surfaces. Although...... important advances were made in the identification of the genetic and extracellular factors required for biofilm formation, the mechanisms leading to biofilm matrix assembly, and the roles of extracellular proteins in these processes are still poorly understood. The symbiont Rhizobium leguminosarum requires...... the synthesis of the acidic exopolysaccharide and the PrsDE secretion system to develop a mature biofilm. PrsDE is responsible for the secretion of the Rap family of proteins that share one or two Ra/CHDL (cadherin-like-) domains. RapA2 is a calcium-dependent lectin with a cadherin-like β sheet structure...

  6. Identification and Characterization of RibN, a Novel Family of Riboflavin Transporters from Rhizobium leguminosarum and Other Proteobacteria

    Science.gov (United States)

    García Angulo, Víctor A.; Bonomi, Hernán R.; Posadas, Diana M.; Serer, María I.; Torres, Alfredo G.; Zorreguieta, Ángeles

    2013-01-01

    Rhizobia are symbiotic bacteria able to invade and colonize the roots of legume plants, inducing the formation of nodules, where bacteria reduce atmospheric nitrogen (N2) to ammonia (NH3). Riboflavin availability influences the capacity of rhizobia to survive in the rhizosphere and to colonize roots. In this study, we identified the RL1692 gene of Rhizobium leguminosarum downstream of a flavin mononucleotide (FMN) riboswitch. RL1692 encodes a putative transmembrane permease with two EamA domains. The presence of an FMN riboswitch regulating a transmembrane protein is usually observed in riboflavin transporters, suggesting that RL1692 may be involved in riboflavin uptake. The product of RL1692, which we named RibN, is conserved in members of the alpha-, beta-, and gammaproteobacteria and shares no significant identity with any riboflavin transporter previously identified. In this work, we show that RibN is localized in the membrane cellular fraction and its expression is downregulated by riboflavin. By heterologous expression in a Brucella abortus mutant auxotrophic for riboflavin, we demonstrate that RibN possesses flavin transport activity. Similarly, we also demonstrate that RibN orthologues from Ochrobactrum anthropi and Vibrio cholerae (which lacks the FMN riboswitch) are able to transport riboflavin. An R. leguminosarum ribN null mutant exhibited lower nodule occupancy levels in pea plants during symbiosis assays. Thus, we propose that RibN and its homologues belong to a novel family of riboflavin transporters. This work provides the first experimental description of riboflavin transporters in Gram-negative bacteria. PMID:23935051

  7. Bacteria–zinc co-localization implicates enhanced synthesis of cysteine-rich peptides in zinc detoxification when Brassica juncea is inoculated with Rhizobium leguminosarum

    Science.gov (United States)

    Adediran, Gbotemi A; Ngwenya, Bryne T; Mosselmans, J Frederick W; Heal, Kate V

    2016-01-01

    Some plant growth promoting bacteria (PGPB) are enigmatic in enhancing plant growth in the face of increased metal accumulation in plants. Since most PGPB colonize the plant root epidermis, we hypothesized that PGPB confer tolerance to metals through changes in speciation at the root epidermis. We employed a novel combination of fluorophore-based confocal laser scanning microscopic imaging and synchrotron based microscopic X-ray fluorescence mapping with X-ray absorption spectroscopy to characterize bacterial localization, zinc (Zn) distribution and speciation in the roots of Brassica juncea grown in Zn contaminated media (400 mg kg−1 Zn) with the endophytic Pseudomonas brassicacearum and rhizospheric Rhizobium leguminosarum. PGPB enhanced epidermal Zn sequestration relative to PGBP-free controls while the extent of endophytic accumulation depended on the colonization mode of each PGBP. Increased root accumulation of Zn and increased tolerance to Zn was associated predominantly with R. leguminosarum and was likely due to the coordination of Zn with cysteine-rich peptides in the root endodermis, suggesting enhanced synthesis of phytochelatins or glutathione. Our mechanistic model of enhanced Zn accumulation and detoxification in plants inoculated with R. leguminosarum has particular relevance to PGPB enhanced phytoremediation of soils contaminated through mining and oxidation of sulphur-bearing Zn minerals or engineered nanomaterials such as ZnS. PMID:26263508

  8. Seroprevalence of brucellosis and typing of Brucella melitensis biovar 2 in lactating cows in Kuwait

    Directory of Open Access Journals (Sweden)

    Adel El-Gohary

    2016-09-01

    Results: The results showed that the overall seroprevalence of bovine brucellosis was 339 (7.25% by BAPAT, 332 (7.1% by RBPT, and 329 (7.04% by CFT. The results revealed that, 42 (8.6%, 5 (1.4% and 292 (7.6% sera were positive for brucellosis by BAPAT in the cows of Al-Wafra, Al-Kabed and Al-Salebia areas, respectively. Whereas, their respective number and seroreactive cases by RBPT were 39 (8.02%, 5 (1.4% and 288 (7.4%. Similarly, as confirmatory test by CFT, the number and seroreactive cases in these areas were 39 (8.02%, 5 (1.4% and 285 (7.46%. MRT revealed that the average positive case was 61.67% (59.46% in Al-Wafra; 60% in Al-Kabed and 66.6% in Al-Salebia. Two Brucella isolates could be recovered from the stomach content of the two aborted feti and typed as Brucella melitensis biovar 2. Conclusion: Brucellosis is prevalent among lactating cows in Kuwait. This indicates the potential role of these dairy animals in disseminating and spread of such zoonosis to human. Considering public health significance, appropriate preventive measures are suggestive for combating brucellosis in Kuwait. [J Adv Vet Anim Res 2016; 3(3.000: 229-235

  9. Analysis of iron acquisition and storage-related genes in clinical and non-clinical strains of Yersinia enterocolitica biovar 1A.

    Science.gov (United States)

    Kanaujia, Pawan Kumar; Bajaj, Priyanka; Virdi, Jugsharan Singh

    2015-10-01

    Possession of mechanisms for iron acquisition and its storage enhances the ability of the bacteria to survive in the iron-limiting environment of the host. In this study, 81 strains of Yersinia enterocolitica biovar 1A isolated from various clinical (n = 51) and non-clinical (n = 30) sources were investigated for the presence of the genes related to iron acquisition and storage. Important genes which were present in more than 85% of the strains included hasA, foxA, bfr, bfd, ftnA, and hmsT as well as the fhuCDB, fepBDGCfesfepA, feoAB, yfuABCD, hemPRSTUV, and hmsHFRS gene clusters. Majority of these genes is being reported for the first time in biovar 1A strains and showed significant homology with genes present in the known pathogenic biovars of Y. enterocolitica. However, no significant difference was observed in the distribution of iron acquisition and storage-related genes among clinical and non-clinical biovar 1A strains. Thus, it may be suggested that the presence of iron acquisition and storage-related genes per se might not be responsible for the supposedly better ability of clinical biovar 1A strains to cause infections in humans. However, in the backdrop of this data, the need to undertake functional studies are highly recommended. © 2015 APMIS. Published by John Wiley & Sons Ltd.

  10. PRODUÇÃO, PROPRIEDADES REOLÓGICAS E COMPOSIÇÃO QUÍMICA DA XANTANA PRODUZIDA POR Xanthomonas axonopodis pv. phaseoli

    OpenAIRE

    Mayer, Lauri; Vendruscolo, Claire Tondo; Silva, Wladimir Padilha da; Moura, Andrea Bittencourt

    2008-01-01

    Goma xantana é um heteropolissacarídeo bacteriano sintetizado por várias espécies do gênero Xanthomonas. Devido principalmente às suas excelentes propriedades reológicas, encontra diversas aplicações na indústria alimentícia, petrolífera, cosmética e em produtos farmacêuticos. Este trabalho objetivou a produção, a análise da viscosidade, do comportamento reológico e a composição química da goma xantana produzida por sete cepas de Xanthomonas axonopodis pv. phaseoli. A produção de xantana vari...

  11. Global survey of mRNA levels and decay rates of Chlamydia trachomatis trachoma and lymphogranuloma venereum biovars

    Directory of Open Access Journals (Sweden)

    Rita Ferreira

    2017-07-01

    Full Text Available Interpreting the intricate bacterial transcriptomics implies understanding the dynamic relationship established between de novo transcription and the degradation of transcripts. Here, we performed a comparative overview of gene expression levels and mRNA decay rates for different-biovar (trachoma and lymphogranuloma venereum strains of the obligate intracellular bacterium Chlamydia trachomatis. By using RNA-sequencing to measure gene expression levels at mid developmental stage and mRNA decay rates upon rifampicin-based transcription blockage, we observed that: i 60–70% of the top-50 expressed genes encode proteins with unknown function and proteins involved in “Translation, ribosomal structure and biogenesis” for all strains; ii the expression ranking by genes' functional categories was in general concordant among different-biovar strains; iii the median of the half-life time (t1/2 values of transcripts were 15–17 min, indicating that the degree of transcripts’ stability seems to correlate with the bacterial intracellular life-style, as these values are considerably higher than the ones observed in other studies for facultative intracellular and free-living bacteria; iv transcript decay rates were highly heterogeneous within each C. trachomatis strain and did not correlate with steady-state expression levels; v only at very few instances (essentially at gene functional category level was possible to unveil dissimilarities potentially underlying phenotypic differences between biovars. In summary, the unveiled transcriptomic scenario, marked by a general lack of correlation between transcript production and degradation and a huge inter-transcript heterogeneity in decay rates, likely reflects the challenges underlying the unique biphasic developmental cycle of C. trachomatis and its intricate interactions with the human host, which probably exacerbate the complexity of the bacterial transcription regulation.

  12. Global survey of mRNA levels and decay rates of Chlamydia trachomatis trachoma and lymphogranuloma venereum biovars.

    Science.gov (United States)

    Ferreira, Rita; Borges, Vítor; Borrego, Maria José; Gomes, João Paulo

    2017-07-01

    Interpreting the intricate bacterial transcriptomics implies understanding the dynamic relationship established between de novo transcription and the degradation of transcripts. Here, we performed a comparative overview of gene expression levels and mRNA decay rates for different-biovar (trachoma and lymphogranuloma venereum) strains of the obligate intracellular bacterium Chlamydia trachomatis . By using RNA-sequencing to measure gene expression levels at mid developmental stage and mRNA decay rates upon rifampicin-based transcription blockage, we observed that: i ) 60-70% of the top-50 expressed genes encode proteins with unknown function and proteins involved in "Translation, ribosomal structure and biogenesis" for all strains; ii ) the expression ranking by genes' functional categories was in general concordant among different-biovar strains; iii ) the median of the half-life time (t 1/2 ) values of transcripts were 15-17 min, indicating that the degree of transcripts' stability seems to correlate with the bacterial intracellular life-style, as these values are considerably higher than the ones observed in other studies for facultative intracellular and free-living bacteria; iv ) transcript decay rates were highly heterogeneous within each C. trachomatis strain and did not correlate with steady-state expression levels; v ) only at very few instances (essentially at gene functional category level) was possible to unveil dissimilarities potentially underlying phenotypic differences between biovars. In summary, the unveiled transcriptomic scenario, marked by a general lack of correlation between transcript production and degradation and a huge inter-transcript heterogeneity in decay rates, likely reflects the challenges underlying the unique biphasic developmental cycle of C. trachomatis and its intricate interactions with the human host, which probably exacerbate the complexity of the bacterial transcription regulation.

  13. First isolation, identification, phenotypic and genotypic characterization of Brucella abortus biovar 3 from dairy cattle in Tanzania.

    Science.gov (United States)

    Mathew, C; Stokstad, M; Johansen, T B; Klevar, S; Mdegela, R H; Mwamengele, G; Michel, P; Escobar, L; Fretin, D; Godfroid, J

    2015-07-21

    Brucellosis is a disease of worldwide public health and economic importance. Successful control is based on knowledge of epidemiology and strains present in an area. In developing countries, most investigations are based on serological assays. This study aimed at investigating a dairy herd experiencing abortions in order to establish within-herd seroprevalence to Brucella spp., identify, characterize Brucella strains by Multiple Loci Variable Number of Tandem Repeats Analysis (MLVA-VNTR) and investigate possible spillover to other species. The within-herd seroprevalence in cattle (n = 200) was 48 % (95 % CI 41-55), using an indirect ELISA, while the Rose Bengal Test (RBT) yielded lower prevalence (21.5 %; 95 % CI 16-27). Two sheep (n = 35) and one goat (n = 50) were seropositive using ELISA while none of the dogs (n = 6) was positive with the RBT. Three Brucella were isolated from an aborted fetus and associated membranes. Real time PCR (IS711), Bruce-ladder and classical biotyping classified the isolates as B. abortus biovar 3. MLVA-VNTR revealed two different but closely related genotypes. The isolates showed unique profiles, providing the first genotypic data from Tanzania. These genotypes were not related to B. abortus biovar 3 reference strain Tulya originally isolated from a human patient in Uganda in 1958, unlike the genotypes isolated and characterized recently in Kenya. High within-herd prevalence, isolation of the pathogen and abortion confirm that B. abortus is circulating in this herd with cattle as reservoir hosts. A low seroprevalence in sheep and goats suggests a spillover of B. abortus from cattle to small ruminants in the herd. This is the first isolation and characterization of B. abortus biovar 3 from a dairy cow with abortion in Tanzania. The origin of the Tanzanian genotypes remain elusive, although they seem to be related to genotypes found in Europe, Turkey and China but not related to B. abortus biovar 3 reference strain or genotypes from

  14. Exploring the Diversity of Field Strains of Brucella abortus Biovar 3 Isolated in West Africa

    Directory of Open Access Journals (Sweden)

    Moussa Sanogo

    2017-06-01

    Full Text Available Brucellosis is one of the most widespread bacterial zoonotic diseases in the world, affecting both humans and domestic and wild animals. Identification and biotyping of field strains of Brucella are of key importance for a better knowledge of the epidemiology of brucellosis, for identifying appropriate antigens, for managing disease outbreaks and for setting up efficient preventive and control programmes. Such data are required both at national and regional level to assess potential threats for public health. Highly discriminative genotyping methods such as the multiple locus variable number of tandem repeats analysis (MLVA allow the comparison and assessment of genetic relatedness between field strains of Brucella within the same geographical area. In this study, MLVA biotyping data retrieved from the literature using a systematic review were compared using a clustering analysis and the Hunter-Gaston diversity index (HGDI. Thus, the analysis of the 42 MLVA genotyping results found in the literature on West Africa [i.e., from Ivory Coast (1, Niger (1, Nigeria (34, The Gambia (3, and Togo (3] did not allow a complete assessment of the actual diversity among field strains of Brucella. However, it provided some preliminary indications on the co-existence of 25 distinct genotypes of Brucella abortus biovar 3 in this region with 19 genotypes from Nigeria, three from Togo and one from Ivory Coast, The Gambia, and Niger. The strong and urgent need for more sustainable molecular data on prevailing strains of Brucella in this sub-region of Africa and also on all susceptible species including humans is therefore highlighted. This remains a necessary stage to allow a comprehensive understanding of the relatedness between field strains of Brucella and the epidemiology of brucellosis within West Africa countries.

  15. Comparative Transcriptome Analysis Reveals Cool Virulence Factors of Ralstonia solanacearum Race 3 Biovar 2.

    Science.gov (United States)

    Meng, Fanhong; Babujee, Lavanya; Jacobs, Jonathan M; Allen, Caitilyn

    2015-01-01

    While most strains of the plant pathogenic bacterium Ralstonia solanacearum are tropical, the race 3 biovar 2 (R3bv2) subgroup attacks plants in cooler climates. To identify mechanisms underlying this trait, we compared the transcriptional profiles of R. solanacearum R3bv2 strain UW551 and tropical strain GMI1000 at 20°C and 28°C, both in culture and during tomato pathogenesis. 4.2% of the ORFs in the UW551 genome and 7.9% of the GMI1000 ORFs were differentially expressed by temperature in planta. The two strains had distinct transcriptional responses to temperature change. GMI1000 up-regulated several stress response genes at 20°C, apparently struggling to cope with plant defenses. At the cooler temperature, R3bv2 strain UW551 up-regulated a cluster encoding a mannose-fucose binding lectin, LecM; a quorum sensing-dependent protein, AidA; and a related hypothetical protein, AidC. The last two genes are absent from the GMI1000 genome. In UW551, all three genes were positively regulated by the adjacent SolI/R quorum sensing system. These temperature-responsive genes were required for full virulence in R3bv2. Mutants lacking lecM, aidA, or aidC were each significantly more reduced in virulence on tomato at 20°C than at 28°C in both a naturalistic soil soak inoculation assay and when they were inoculated directly into tomato stems. The lecM and aidC mutants also survived poorly in potato tubers at the seed tuber storage temperature of 4°C, and the lecM mutant was defective in biofilm formation in vitro. Together, these results suggest novel mechanisms, including a lectin, are involved in the unique temperate epidemiology of R3bv2.

  16. Identification and characterization of PhoP regulon members in Yersinia pestis biovar Microtus

    Directory of Open Access Journals (Sweden)

    Du Zongmin

    2008-03-01

    Full Text Available Abstract Background The transcription regulator PhoP has been shown to be important for Y. pestis survival in macrophages and under various in vitro stresses. However, the mechanism by which PhoP promotes bacterial intracellular survival is not fully understood. Our previous microarray analysis suggested that PhoP governed a wide set of cellular pathways in Y. pestis. A series of biochemical experiments were done herein to study members of the PhoP regulon of Y. pestis biovar Microtus. Results By using gel mobility shift assay and quantitative RT-PCR, a total of 30 putative transcription units were characterized as direct PhoP targets. The primer extension assay was further used to determine the transcription start sites of 18 PhoP-dependent promoters and to localize the -10 and -35 elements. The DNase I footprinting was used to identify the PhoP-binding sites within 17 PhoP-dependent promoters, enabling the identification of PhoP box and matrix that both represented the conserved signals for PhoP recognition in Y. pestis. Data presented here providing a good basis for modeling PhoP-promoter DNA interactions that is crucial to the PhoP-mediated transcriptional regulation. Conclusion The proven direct PhoP targets include nine genes encoding regulators and 21 genes or operons with functions of detoxification, protection against DNA damages, resistance to antimicrobial peptides, and adaptation to magnesium limitation. We can presume that PhoP is a global regulator that controls a complex regulatory cascade by a mechanism of not only directly controlling the expression of specific genes, but also indirectly regulating various cellular pathways by acting on a set of dedicated regulators. These results help us gain insights into the PhoP-dependent mechanisms by which Y. pestis survives the antibacterial strategies employed by host macrophages.

  17. Comparative Transcriptome Analysis Reveals Cool Virulence Factors of Ralstonia solanacearum Race 3 Biovar 2.

    Directory of Open Access Journals (Sweden)

    Fanhong Meng

    Full Text Available While most strains of the plant pathogenic bacterium Ralstonia solanacearum are tropical, the race 3 biovar 2 (R3bv2 subgroup attacks plants in cooler climates. To identify mechanisms underlying this trait, we compared the transcriptional profiles of R. solanacearum R3bv2 strain UW551 and tropical strain GMI1000 at 20°C and 28°C, both in culture and during tomato pathogenesis. 4.2% of the ORFs in the UW551 genome and 7.9% of the GMI1000 ORFs were differentially expressed by temperature in planta. The two strains had distinct transcriptional responses to temperature change. GMI1000 up-regulated several stress response genes at 20°C, apparently struggling to cope with plant defenses. At the cooler temperature, R3bv2 strain UW551 up-regulated a cluster encoding a mannose-fucose binding lectin, LecM; a quorum sensing-dependent protein, AidA; and a related hypothetical protein, AidC. The last two genes are absent from the GMI1000 genome. In UW551, all three genes were positively regulated by the adjacent SolI/R quorum sensing system. These temperature-responsive genes were required for full virulence in R3bv2. Mutants lacking lecM, aidA, or aidC were each significantly more reduced in virulence on tomato at 20°C than at 28°C in both a naturalistic soil soak inoculation assay and when they were inoculated directly into tomato stems. The lecM and aidC mutants also survived poorly in potato tubers at the seed tuber storage temperature of 4°C, and the lecM mutant was defective in biofilm formation in vitro. Together, these results suggest novel mechanisms, including a lectin, are involved in the unique temperate epidemiology of R3bv2.

  18. A model for the catabolism of rhizopine in Rhizobium leguminosarum involves a ferredoxin oxygenase complex and the inositol degradative pathway.

    Science.gov (United States)

    Bahar, M; de Majnik, J; Wexler, M; Fry, J; Poole, P S; Murphy, P J

    1998-11-01

    Rhizopines are nodule-specific compounds that confer an intraspecies competitive nodulation advantage to strains that can catabolize them. The rhizopine (3-O-methyl-scyllo-inosamine, 3-O-MSI) catabolic moc gene cluster mocCABRDE(F) in Rhizobium leguminosarum bv. viciae strain 1a is located on the Sym plasmid. MocCABR are homologous to the mocCABR gene products from Sinorhizobium meliloti. MocD and MocE contain motifs corresponding to a TOL-like oxygenase and a [2Fe-2S] Rieske-like ferredoxin, respectively. The mocF gene encodes a ferredoxin reductase that would complete the oxygenase system, but is not essential for rhizopine catabolism. We propose a rhizopine catabolic model whereby MocB transports rhizopine into the cell and MocDE and MocF (or a similar protein elsewhere in the genome), under the regulation of MocR, act in concert to form a ferredoxin oxygenase system that demethylates 3-O-MSI to form scyllo-inosamine (SI). MocA, an NAD(H)-dependent dehydrogenase, and MocC continue the catabolic process. Compounds formed then enter the inositol catabolic pathway.

  19. Characterisation of SalRAB a salicylic acid inducible positively regulated efflux system of Rhizobium leguminosarum bv viciae 3841.

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    Adrian J Tett

    Full Text Available Salicylic acid is an important signalling molecule in plant-microbe defence and symbiosis. We analysed the transcriptional responses of the nitrogen fixing plant symbiont, Rhizobium leguminosarum bv viciae 3841 to salicylic acid. Two MFS-type multicomponent efflux systems were induced in response to salicylic acid, rmrAB and the hitherto undescribed system salRAB. Based on sequence similarity salA and salB encode a membrane fusion and inner membrane protein respectively. salAB are positively regulated by the LysR regulator SalR. Disruption of salA significantly increased the sensitivity of the mutant to salicylic acid, while disruption of rmrA did not. A salA/rmrA double mutation did not have increased sensitivity relative to the salA mutant. Pea plants nodulated by salA or rmrA strains did not have altered nodule number or nitrogen fixation rates, consistent with weak expression of salA in the rhizosphere and in nodule bacteria. However, BLAST analysis revealed seventeen putative efflux systems in Rlv3841 and several of these were highly differentially expressed during rhizosphere colonisation, host infection and bacteroid differentiation. This suggests they have an integral role in symbiosis with host plants.

  20. Characterization of bacterial isolates from rotting potato tuber tissue showing antagonism to Dickeya sp. biovar 3 in vitro and in planta

    NARCIS (Netherlands)

    Czajkowski, R.L.; De Boer, W.J.; Van Veen, J.A.; Van der Wolf, J.M.

    2012-01-01

    Possibilities for biocontrol of biovar 3 Dickeya sp. in potato were investigated, using bacteria from rotting potato tissue isolated by dilution plating on nonselective agar media. In a plate assay, 649 isolates were screened for antibiosis against Dickeya sp. IPO2222 and for the production of

  1. Studies on the interaction between the biocontrol agent, Serratia plymuthica A30, and blackleg-causing Dickeya sp. (biovar 3) in potato (Solanum tuberosum)

    NARCIS (Netherlands)

    Czajkowski, R.L.; Boer, de W.J.; Veen, van J.A.; Wolf, van der J.M.

    2012-01-01

    Interactions between Serratia plymuthica A30 and a blackleg-causing biovar 3 Dickeya sp. were examined. In a potato slice assay, S. plymuthica A30 inhibited tissue maceration caused by Dickeya sp. IPO2222 when co-inoculated at a density at least 10 times greater than that of the pathogen. In

  2. Studies on the interaction between the biocontrol agent, Serratia plymuthica A30, and blackleg-causing Dickeya sp (biovar 3) in potato (Solanum tuberosum)

    NARCIS (Netherlands)

    Czajkowski, R.L.; De Boer, W.J.; Van Veen, J.A.; Van der Wolf, J.M.

    2012-01-01

    Interactions between Serratia plymuthica A30 and a blackleg-causing biovar 3 Dickeya sp. were examined. In a potato slice assay, S. plymuthica A30 inhibited tissue maceration caused by Dickeya sp. IPO2222 when co-inoculated at a density at least 10 times greater than that of the pathogen. In

  3. Nucleotide Sequence and Analysis of an orotate transporter-containing plasmid isolated from the Lactococcus lactis ssp. lactis biovar diacetylactis strain DB0410

    DEFF Research Database (Denmark)

    Defoor, Els Marie Celine; Martinussen, Jan

    A new lactococcal plasmid, pDBORO, was isolated from the Lactococcus lactis ssp. lactis biovar diacetylactis strain DB0410 responsible for the sensitivity of DB0410 towards the pyrimidine-analog 5´-fluoroorotate. The plasmid pDBORO amounts to 16404 bp and its complete nucleotide sequence has been...

  4. Mutation of praR in Rhizobium leguminosarum enhances root biofilms, improving nodulation competitiveness by increased expression of attachment proteins.

    Science.gov (United States)

    Frederix, Marijke; Edwards, Anne; Swiderska, Anna; Stanger, Andrew; Karunakaran, Ramakrishnan; Williams, Alan; Abbruscato, Pamela; Sanchez-Contreras, Maria; Poole, Philip S; Downie, J Allan

    2014-08-01

    In Rhizobium leguminosarum bv. viciae, quorum-sensing is regulated by CinR, which induces the cinIS operon. CinI synthesizes an AHL, whereas CinS inactivates PraR, a repressor. Mutation of praR enhanced biofilms in vitro. We developed a light (lux)-dependent assay of rhizobial attachment to roots and demonstrated that mutation of praR increased biofilms on pea roots. The praR mutant out-competed wild-type for infection of pea nodules in mixed inoculations. Analysis of gene expression by microarrays and promoter fusions revealed that PraR represses its own transcription and mutation of praR increased expression of several genes including those encoding secreted proteins (the adhesins RapA2, RapB and RapC, two cadherins and the glycanase PlyB), the polysaccharide regulator RosR, and another protein similar to PraR. PraR bound to the promoters of several of these genes indicating direct repression. Mutations in rapA2, rapB, rapC, plyB, the cadherins or rosR did not affect the enhanced root attachment or nodule competitiveness of the praR mutant. However combinations of mutations in rapA, rapB and rapC abolished the enhanced attachment and nodule competitiveness. We conclude that relief of PraR-mediated repression determines a lifestyle switch allowing the expression of genes that are important for biofilm formation on roots and the subsequent initiation of infection of legume roots. © 2014 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.

  5. Functional relationships between plasmids and their significance for metabolism and symbiotic performance of Rhizobium leguminosarum bv. trifolii.

    Science.gov (United States)

    Stasiak, Grażyna; Mazur, Andrzej; Wielbo, Jerzy; Marczak, Małgorzata; Zebracki, Kamil; Koper, Piotr; Skorupska, Anna

    2014-11-01

    Rhizobium leguminosarum bv. trifolii TA1 (RtTA1) is a soil bacterium establishing a highly specific symbiotic relationship with clover, which is based on the exchange of molecular signals between the host plant and the microsymbiont. The RtTA1 genome is large and multipartite, composed of a chromosome and four plasmids, which comprise approximately 65 % and 35 % of the total genome, respectively. Extrachromosomal replicons were previously shown to confer significant metabolic versatility to bacteria, which is important for their adaptation in the soil and nodulation competitiveness. To investigate the contribution of individual RtTA1 plasmids to the overall cell phenotype, metabolic properties and symbiotic performance, a transposon-based elimination strategy was employed. RtTA1 derivatives cured of pRleTA1b or pRleTA1d and deleted in pRleTA1a were obtained. In contrast to the in silico predictions of pRleTA1b and pRleTA1d, which were described as chromid-like replicons, both appeared to be completely curable. On the other hand, for pRleTA1a (symbiotic plasmid) and pRleTA1c, which were proposed to be unessential for RtTA1 viability, it was not possible to eliminate them at all (pRleTA1c) or entirely (pRleTA1a). Analyses of the phenotypic traits of the RtTA1 derivatives obtained revealed the functional significance of individual plasmids and their indispensability for growth, certain metabolic pathways, production of surface polysaccharides, autoaggregation, biofilm formation, motility and symbiotic performance. Moreover, the results allow us to suggest broad functional cooperation among the plasmids in shaping the phenotypic properties and symbiotic capabilities of rhizobia.

  6. Structural Variabilities in β-Lactamase (blaA of Different Biovars of Yersinia enterocolitica: Implications for β-Lactam Antibiotic and β-Lactamase Inhibitor Susceptibilities.

    Directory of Open Access Journals (Sweden)

    Neelja Singhal

    Full Text Available Yersiniosis caused by Yersinia enterocolitica has been reported from all continents. The bacterial species is divided into more than fifty serovars and six biovars viz. 1A, 1B, 2, 3, 4 and 5 which differ in geographical distribution, ecological niches and pathogenicity. Most Y.enterocolitica strains harbor chromosomal genes for two β-lactamases, blaA an Ambler class A penicillinase and blaB an Ambler class C inducible cephalosporinase. In the present study, susceptibility to b-lactam antibiotics and β-lactamase inhibitor was studied for Y. enterocolitica strains of biovars 1A, 1B, 2 and 4. We observed that β-lactamases were expressed differentially among strains of different biovars. To understand the molecular mechanisms underlying such differential expression, the sequences of genes and promoters of blaA were compared. Also, the variants of blaA present in different biovars were modeled and docked with amoxicillin and clavulanic acid. The mRNA secondary structures of blaA variants were also predicted in-silico. Our findings indicated that neither variations in the promoter regions, nor the secondary structures of mRNA contributed to higher/lower expression of blaA in different biovars. Analysis of H-bonding residues of blaA variants with amoxicillin and clavulanic acid revealed that if amino acid residues of a β-lactamase interacting with amoxicillin and the clavulanic acid were similar, clavulanic acid was effective in engaging the enzyme, accounting for a significant reduction in MIC of amoxicillin-clavulanate. This finding might aid in designing better β-lactamase inhibitors with improved efficiencies in future.

  7. Epidemiological investigation of the first human brucellosis case in Spain due to Brucella suis biovar 1 strain 1330.

    Science.gov (United States)

    Compés Dea, Cecilia; Guimbao Bescós, Joaquín; Alonso Pérez de Ágreda, Juan Pablo; Muñoz Álvaro, Pilar María; Blasco Martínez, José María; Villuendas Usón, María Cruz

    2017-03-01

    No cases of human brucellosis caused by Brucella suis has been reported in Spain. This study involved interviews with the case and his co-workers, inspection of their workplace, checking infection control measures, and typing the Brucella strain isolated in the blood culture. Brucella suis biovar 1 strain 1330 was isolated from a patient who worked in a waste treatment plant. Food borne transmission, contact with animals, and risk jobs were ruled out. An accidental inoculation with a contaminated needle from a research laboratory waste container was identified as the most probable mode of transmission. There should be controls to ensure that waste containers are sealed. Copyright © 2016 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.

  8. Effect of domestic cooking on the starch digestibility, predicted glycemic indices, polyphenol contents and alpha amylase inhibitory properties of beans (Phaseolis vulgaris) and breadfruit (Treculia africana).

    Science.gov (United States)

    Chinedum, E; Sanni, S; Theressa, N; Ebere, A

    2018-01-01

    The effect of processing on starch digestibility, predicted glycemic indices (pGI), polyphenol contents and alpha amylase inhibitory properties of beans (Phaseolis vulgaris) and breadfruit (Treculia africana) was studied. Total starch ranged from 4.3 to 68.3g/100g, digestible starch ranged from 4.3 to 59.2 to 65.7g/100g for the raw and processed legumes; Resistance starch was not detected in most of the legumes except in fried breadfruit and the starches in both the raw and processed breadfruit were more rapidly digested than those from raw and cooked beans. Raw and processed breadfruit had higher hydrolysis curves than raw and processed beans with the amylolysis level in raw breadfruit close to that of white bread. Raw beans had a low glycemic index (GI); boiled beans and breadfruit had intermediate glycemic indices respectively while raw and fried breadfruit had high glycemic indices. Aqueous extracts of the food samples had weak α-amylase inhibition compared to acarbose. The raw and processed legumes contained considerable amounts of dietary phenols and flavonoids. The significant correlation (r=0.626) between α-amylase inhibitory actions of the legumes versus their total phenolic contents suggests the contribution of the phenolic compounds in these legumes to their α-amylase inhibitory properties. Copyright © 2017 Elsevier B.V. All rights reserved.

  9. Production of inulinase by Xanthomonas campestris pv phaseoli using onion (Allium cepa) and garlic (Allium sativum) peels in solid state cultivation.

    Science.gov (United States)

    Ayyachamy, M; Khelawan, K; Pillay, D; Permaul, K; Singh, S

    2007-10-01

    To access inulinase production by Xanthomonas campestris pv phaseoli using the submerged and solid state cultivation (SSC) methods. Various carbon sources, inulin-rich solid substrates and pure synthetic inulin were tested for their efficiency in inulinase induction. The highest inulinase production (17.42 IU ml(-1)) in submerged cultures of X. campestris was observed with inulin as a carbon source with an initial pH, temperature and agitation of 7.0, 37 degrees C and 150 rev min(-1) respectively. Among the various substrates, garlic peels (117 IU gds(-1)) and onion peels (101 IU gds(-1)) were found to be the best for inulinase production. The inulinase production level of X. campestris was 6.7-fold higher in garlic and 5.8-fold in onion, under optimized SSC conditions compared with the submerged culture. This is the first report on inulinase production from garlic and onion peels by X. campestris using SSC. SSC is an efficient method for inulinase production by X. campestris for commercial applications.

  10. Assessment of the genetic and phenotypic diversity among rhizogenic Agrobacterium biovar 1 strains infecting solanaceous and cucurbit crops.

    Science.gov (United States)

    Bosmans, Lien; Álvarez-Pérez, Sergio; Moerkens, Rob; Wittemans, Lieve; Van Calenberge, Bart; Kerckhove, Stefan Van; Paeleman, Anneleen; De Mot, René; Rediers, Hans; Lievens, Bart

    2015-08-01

    Rhizogenic Agrobacterium biovar 1 strains have been found to cause extensive root proliferation on hydroponically grown Cucurbitaceae and Solanaceae crops, resulting in substantial economic losses. As these agrobacteria live under similar ecological conditions, infecting a limited number of crops, it may be hypothesized that genetic and phenotypic variation among such strains is relatively low. In this study we assessed the phenotypic diversity as well as the phylogenetic and evolutionary relationships of several rhizogenic Agrobacterium biovar 1 strains from cucurbit and solanaceous crops. A collection of 41 isolates was subjected to a number of phenotypic assays and characterized by MLSA targeting four housekeeping genes (16S rRNA gene, recA, rpoB and trpE) and two loci from the root-inducing Ri-plasmid (part of rolB and virD2). Besides phenotypic variation, remarkable genotypic diversity was observed, especially for some chromosomal loci such as trpE. In contrast, genetic diversity was lower for the plasmid-borne loci, indicating that the studied chromosomal housekeeping genes and Ri-plasmid-borne loci might not exhibit the same evolutionary history. Furthermore, phylogenetic and network analyses and several recombination tests suggested that recombination could be contributing in some extent to the evolutionary dynamics of rhizogenic Agrobacterium populations. Finally, a genomospecies-level identification analysis revealed that at least four genomospecies may occur on cucurbit and tomato crops (G1, G3, G8 and G9). Together, this study gives a first glimpse at the genetic and phenotypic diversity within this economically important plant pathogenic bacterium. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  11. Genomic Structural Variations Affecting Virulence During Clonal Expansion of Pseudomonas syringae pv. actinidiae Biovar 3 in Europe.

    Science.gov (United States)

    Firrao, Giuseppe; Torelli, Emanuela; Polano, Cesare; Ferrante, Patrizia; Ferrini, Francesca; Martini, Marta; Marcelletti, Simone; Scortichini, Marco; Ermacora, Paolo

    2018-01-01

    Pseudomonas syringae pv. actinidiae (Psa) biovar 3 caused pandemic bacterial canker of Actinidia chinensis and Actinidia deliciosa since 2008. In Europe, the disease spread rapidly in the kiwifruit cultivation areas from a single introduction. In this study, we investigated the genomic diversity of Psa biovar 3 strains during the primary clonal expansion in Europe using single molecule real-time (SMRT), Illumina and Sanger sequencing technologies. We recorded evidences of frequent mobilization and loss of transposon Tn6212, large chromosome inversions, and ectopic integration of IS sequences (remarkably ISPsy31, ISPsy36, and ISPsy37). While no phenotype change associated with Tn6212 mobilization could be detected, strains CRAFRU 12.29 and CRAFRU 12.50 did not elicit the hypersensitivity response (HR) on tobacco and eggplant leaves and were limited in their growth in kiwifruit leaves due to insertion of ISPsy31 and ISPsy36 in the hrpS and hrpR genes, respectively, interrupting the hrp cluster. Both strains had been isolated from symptomatic plants, suggesting coexistence of variant strains with reduced virulence together with virulent strains in mixed populations. The structural differences caused by rearrangements of self-genetic elements within European and New Zealand strains were comparable in number and type to those occurring among the European strains, in contrast with the significant difference in terms of nucleotide polymorphisms. We hypothesize a relaxation, during clonal expansion, of the selection limiting the accumulation of deleterious mutations associated with genome structural variation due to transposition of mobile elements. This consideration may be relevant when evaluating strategies to be adopted for epidemics management.

  12. Draft Genome Sequence of Highly Virulent Race 4/Biovar 3 of Ralstonia solanacearum CaRs_Mep Causing Bacterial Wilt in Zingiberaceae Plants in India.

    Science.gov (United States)

    Kumar, Aundy; Munjal, Vibhuti; Sheoran, Neelam; Prameela, Thekkan Puthiyaveedu; Suseelabhai, Rajamma; Aggarwal, Rashmi; Jain, Rakesh Kumar; Eapen, Santhosh J

    2017-01-05

    The genome of Ralstonia solanacearum CaRs_Mep, a race 4/biovar 3/phylotype I bacterium causing wilt in small cardamom and other Zingiberaceae plants, was sequenced. Analysis of the 5.7-Mb genome sequence will aid in better understanding of the genetic determinants of host range, host jump, survival, pathogenicity, and virulence of race 4 of R. solanacearum. Copyright © 2017 Kumar et al.

  13. Genetic relationships between clinical and non-clinical strains of Yersinia enterocolitica biovar 1A as revealed by multilocus enzyme electrophoresis and multilocus restriction typing

    Directory of Open Access Journals (Sweden)

    Virdi Jugsharan S

    2010-05-01

    Full Text Available Abstract Background Genetic relationships among 81 strains of Y. enterocolitica biovar 1A isolated from clinical and non-clinical sources were discerned by multilocus enzyme electrophoresis (MLEE and multilocus restriction typing (MLRT using six loci each. Such studies may reveal associations between the genotypes of the strains and their sources of isolation. Results All loci were polymorphic and generated 62 electrophoretic types (ETs and 12 restriction types (RTs. The mean genetic diversity (H of the strains by MLEE and MLRT was 0.566 and 0.441 respectively. MLEE (DI = 0.98 was more discriminatory and clustered Y. enterocolitica biovar 1A strains into four groups, while MLRT (DI = 0.77 identified two distinct groups. BURST (Based Upon Related Sequence Types analysis of the MLRT data suggested aquatic serotype O:6,30-6,31 isolates to be the ancestral strains from which, clinical O:6,30-6,31 strains might have originated by host adaptation and genetic change. Conclusion MLEE revealed greater genetic diversity among strains of Y. enterocolitica biovar 1A and clustered strains in four groups, while MLRT grouped the strains into two groups. BURST analysis of MLRT data nevertheless provided newer insights into the probable evolution of clinical strains from aquatic strains.

  14. Evaluation Of GAMMA And Sodium Azide Radiation Looking For Resistance To MACROPHOMINA Phaseoli T. In Three Promissory Lines Of Benne Sesamum Indicum L

    International Nuclear Information System (INIS)

    Lopez R, Liliana V.; Melo O, Mario E.; Reina C, Yasmin

    1993-01-01

    The Benne in Colombia is a cultivation relegated to small properties and its main problem is the withering (Macrophomina phaseoli T.) which may cause losses of the cent per cent. Situation that generated the interest to carry out a study on induction of mutations in this species, using Gamma Rays and Sodium Azide with the purpose of obtaining possible resistant mutants to the mushroom. The initial stage of the work, determination of good dose, was developed in the INEA in Bogota and the field evaluation in the ICA Nataima in Espinal. A rehearsal was mounted to evaluate germination and plants height in the lines Cycle 0 - PlO, ENA 841-3 and ENA 841-5 tried with the doses (OR, 25, 50, 75 and 100) krad of gamma rays and (OR, 5, 10, 15 and 20) mm of Sodium Azide, the obtained information was processed using a Complete Blocks design at random with Factorial arrangement 3x5 and test of Duncan, based on the obtained data the Curves of Radio and chemical sensitivity were determined. Then the massive treatment of the seed was made with the optimum mutagenic dose and it was taken to field (Ml), giving it good conditions for the development of the cultivation. The test of chlorophyll ian mutants was applied to the seed harvested M2 to select the treatment with which the program of improvement should continue, it was sowed in a lot with high pressure of selection of the mushroom and an inter and intra family selection was made

  15. Differentially Expressed Genes in Resistant and Susceptible Common Bean (Phaseolus vulgaris L. Genotypes in Response to Fusarium oxysporum f. sp. phaseoli.

    Directory of Open Access Journals (Sweden)

    Renfeng Xue

    Full Text Available Fusarium wilt of common bean (Phaseolus vulgaris L., caused by Fusarium oxysporum Schlechtend.:Fr. f.sp. phaseoli (Fop, is one of the most important diseases of common beans worldwide. Few natural sources of resistance to Fop exist and provide only moderate or partial levels of protection. Despite the economic importance of the disease across multiple crops, only a few of Fop induced genes have been analyzed in legumes. Therefore, our goal was to identify transcriptionally regulated genes during an incompatible interaction between common bean and the Fop pathogen using the cDNA amplified fragment length polymorphism (cDNA-AFLP technique. We generated a total of 8,730 transcript-derived fragments (TDFs with 768 primer pairs based on the comparison of a moderately resistant and a susceptible genotype. In total, 423 TDFs (4.9% displayed altered expression patterns after inoculation with Fop inoculum. We obtained full amplicon sequences for 122 selected TDFs, of which 98 were identified as annotated known genes in different functional categories based on their putative functions, 10 were predicted but non-annotated genes and 14 were not homologous to any known genes. The 98 TDFs encoding genes of known putative function were classified as related to metabolism (22, signal transduction (21, protein synthesis and processing (20, development and cytoskeletal organization (12, transport of proteins (7, gene expression and RNA metabolism (4, redox reactions (4, defense and stress responses (3, energy metabolism (3, and hormone responses (2. Based on the analyses of homology, 19 TDFs from different functional categories were chosen for expression analysis using quantitative RT-PCR. The genes found to be important here were implicated at various steps of pathogen infection and will allow a better understanding of the mechanisms of defense and resistance to Fop and similar pathogens. The differential response genes discovered here could also be used as

  16. The Dry Plant Extract of Common Bean Seed (Phaseoli Vulgari Pericarpium does not Have an Affect on Postprandial Glycemia in Healthy Human Subject

    Directory of Open Access Journals (Sweden)

    Aleksandra Cerović

    2006-08-01

    Full Text Available The aim of present study was to assess the effects and safety of a dry Phaseoli vulgari pericarpium (PVP extract on postprandial glycemia in healthy participants. A randomized crossover experiment where participants received either PVP extract or placebo. Chemical compounds in dry extract were assessed by established methods. Eighteen healthy participants (9 male and 9 female aged 29+/-4,8 years, body mass index (BMI 23+/-3,7 kg/m(2 were recruited among students and staff at the Faculty of Pharmacy, University of Belgrade. All participants were able to follow the study protocol without difficulty. The participants received either PVP extract or placebo 30 minutes before a 50g oral glucose tolerance test (OGTT. The protocol followed the guidelines for the OGTT with blood samples drawn at 0, 15, 30, 60, 90 and 120 min. This study demonstrated that there was no significantly effect of the PVP extract on incremental blood glucose (IBG and their areas under the curve (AUC neither male nor female participants. However, IBG together with AUC changes were significantly lower in male compared with female participants in treated and untreated groups. The presence of chrome, soluble fiber, vitamin C, protein, glucose and lectins were also quantified. The applied amount of PVP extract was unable to produce the postprandial hypoglycemia. We assumed that amounts of chrome, soluble fiber, vitamin C which have beneficial effects on diabetes treatment were sufficient to produce hypoglycemia.

  17. Yersinia pestis biovar Microtus strain 201, an avirulent strain to humans, provides protection against bubonic plague in rhesus macaques.

    Science.gov (United States)

    Zhang, Qingwen; Wang, Qiong; Tian, Guang; Qi, Zhizhen; Zhang, Xuecan; Wu, Xiaohong; Qiu, Yefeng; Bi, Yujing; Yang, Xiaoyan; Xin, Youquan; He, Jian; Zhou, Jiyuan; Zeng, Lin; Yang, Ruifu; Wang, Xiaoyi

    2014-01-01

    Yersinia pestis biovar Microtus is considered to be a virulent to larger mammals, including guinea pigs, rabbits and humans. It may be used as live attenuated plague vaccine candidates in terms of its low virulence. However, the Microtus strain's protection against plague has yet to be demonstrated in larger mammals. In this study, we evaluated the protective efficacy of the Microtus strain 201 as a live attenuated plague vaccine candidate. Our results show that this strain is highly attenuated by subcutaneous route, elicits an F1-specific antibody titer similar to the EV and provides a protective efficacy similar to the EV against bubonic plague in Chinese-origin rhesus macaques. The Microtus strain 201 could induce elevated secretion of both Th1-associated cytokines (IFN-γ, IL-2 and TNF-α) and Th2-associated cytokines (IL-4, IL-5, and IL-6), as well as chemokines MCP-1 and IL-8. However, the protected animals developed skin ulcer at challenge site with different severity in most of the immunized and some of the EV-immunized monkeys. Generally, the Microtus strain 201 represented a good plague vaccine candidate based on its ability to generate strong humoral and cell-mediated immune responses as well as its good protection against high dose of subcutaneous virulent Y. pestis challenge.

  18. Bacillus cereus Biovar Anthracis Causing Anthrax in Sub-Saharan Africa-Chromosomal Monophyly and Broad Geographic Distribution.

    Directory of Open Access Journals (Sweden)

    Kym S Antonation

    2016-09-01

    Full Text Available Through full genome analyses of four atypical Bacillus cereus isolates, designated B. cereus biovar anthracis, we describe a distinct clade within the B. cereus group that presents with anthrax-like disease, carrying virulence plasmids similar to those of classic Bacillus anthracis. We have isolated members of this clade from different mammals (wild chimpanzees, gorillas, an elephant and goats in West and Central Africa (Côte d'Ivoire, Cameroon, Central African Republic and Democratic Republic of Congo. The isolates shared several phenotypic features of both B. anthracis and B. cereus, but differed amongst each other in motility and their resistance or sensitivity to penicillin. They all possessed the same mutation in the regulator gene plcR, different from the one found in B. anthracis, and in addition, carry genes which enable them to produce a second capsule composed of hyaluronic acid. Our findings show the existence of a discrete clade of the B. cereus group capable of causing anthrax-like disease, found in areas of high biodiversity, which are possibly also the origin of the worldwide distributed B. anthracis. Establishing the impact of these pathogenic bacteria on threatened wildlife species will require systematic investigation. Furthermore, the consumption of wildlife found dead by the local population and presence in a domestic animal reveal potential sources of exposure to humans.

  19. Resistance to amoxicillin-clavulanate and its relation to virulence-related factors in Yersinia enterocolitica biovar 1A

    Directory of Open Access Journals (Sweden)

    N Singhal

    2016-01-01

    Full Text Available Recent studies have reported that the virulence factors (VFs were detected more frequently in amoxicillin-clavulanate (AMC susceptible clinical isolates of Escherichia coli. Here, we have evaluated the relationship between VFs and AMC-resistance phenotype in clinical isolates of Y. enterocolitica biovar 1A. The presence/absence of VFs was compared with their minimum inhibitory concentrations for AMC in strains of two serovars. We observed that the strains of the serovar O: 6, 30-6, 31 showed a similar relationship between the number of VFs and resistance to clavulanic acid as in E. coli but not of serovar O: 6, 30. Variations in the promoters/complete coding sequences (CCDSs of β-lactamase gene (bla A or the serological characteristics could not account for unusual susceptibility to AMC displayed by the strains of the serovar O: 6, 30. Therefore, we speculate that since the clinical strains of serovar O: 6, 30-6, 31 originated from the environment they were less exposed to antibiotics compared to clinical strains of serovar O: 6, 30. Thus, AMC susceptibility seems to be influenced by factors other than serotypes or promoters/CCDS of β-lactamase genes.

  20. Prevalence and characterization of multi-drug resistant Salmonella Enterica serovar Gallinarum biovar Pullorum and Gallinarum from chicken

    Directory of Open Access Journals (Sweden)

    Md. Shafiullah Parvej

    2016-01-01

    Full Text Available Aim: Salmonella is an important zoonotic pathogen responsible for animal and human diseases. The aim of the present study was to determine the prevalence and stereotyping of Salmonella isolates isolated from apparently healthy poultry. Furthermore, the clonal relatedness among the isolated Salmonella serovars was assessed. Materials and Methods: A total of 150 cloacal swab samples from apparently healthy chickens were collected, and were subjected for the isolation and identification of associated Salmonella organisms. The isolated colonies were identified and characterized on the basis of morphology, cultural characters, biochemical tests, slide agglutination test, polymerase chain reaction, and pulsed-field gel electrophoresis (PFGE. Antibiotic sensitivity patterns were also investigated using commonly used antibiotics. Results: Of the 150 samples, 11 (7.33% produced characteristics pink colony with black center on XLD agar medium, and all were culturally and biochemically confirmed to be Salmonella. All possessed serovar-specific gene SpeF and reacted uniformly with group D antisera, suggesting that all of the isolates were Salmonella Enterica serovar Gallinarum, biovar Pullorum and/or Gallinarum. Antimicrobial susceptibility testing revealed that 54.54% of the isolated Salmonella Enterica serovars were highly sensitive to ciprofloxacin, whereas the 81.81% isolates were resistant to amoxycillin, doxycycline, kanamycin, gentamycin, and tetracycline. Pulsed-field gel electrophoresis of the XbaI-digested genomic DNA exhibited identical banding patterns, suggesting that the multidrug resistant Salmonella Enterica serovars occurring in commercial layers are highly clonal in Bangladesh. Conclusion: The present study was conducted to find out the prevalence of poultry Salmonella in layer chicken and to find out the clonal relationship among them. The data in this study suggest the prevalence of Salmonella Enterica, which is multidrug resistant and

  1. The Lipopolysaccharide Lipid A Long-Chain Fatty Acid Is Important for Rhizobium leguminosarum Growth and Stress Adaptation in Free-Living and Nodule Environments.

    Science.gov (United States)

    Bourassa, Dianna V; Kannenberg, Elmar L; Sherrier, D Janine; Buhr, R Jeffrey; Carlson, Russell W

    2017-02-01

    Rhizobium bacteria live in soil and plant environments, are capable of inducing symbiotic nodules on legumes, invade these nodules, and develop into bacteroids that fix atmospheric nitrogen into ammonia. Rhizobial lipopolysaccharide (LPS) is anchored in the bacterial outer membrane through a specialized lipid A containing a very long-chain fatty acid (VLCFA). VLCFA function for rhizobial growth in soil and plant environments is not well understood. Two genes, acpXL and lpxXL, encoding acyl carrier protein and acyltransferase, are among the six genes required for biosynthesis and transfer of VLCFA to lipid A. Rhizobium leguminosarum mutant strains acpXL, acpXL - /lpxXL - , and lpxXL - were examined for LPS structure, viability, and symbiosis. Mutations in acpXL and lpxXL abolished VLCFA attachment to lipid A. The acpXL mutant transferred a shorter acyl chain instead of VLCFA. Strains without lpxXL neither added VLCFA nor a shorter acyl chain. In all strains isolated from nodule bacteria, lipid A had longer acyl chains compared with laboratory-cultured bacteria, whereas mutant strains displayed altered membrane properties, modified cationic peptide sensitivity, and diminished levels of cyclic β-glucans. In pea nodules, mutant bacteroids were atypically formed and nitrogen fixation and senescence were affected. The role of VLCFA for rhizobial environmental fitness is discussed.

  2. Effect in field of the native strain COL6 of Rhizobium leguminosarum bv. viciae on common bean cv. Percal in Ecuador

    Directory of Open Access Journals (Sweden)

    Klever Iván Granda-Mora

    2017-07-01

    Full Text Available Inoculation with selected Rhizobium strains of common bean (Phaseolus vulgaris L. contributes to the agricultural yield of this legume, improves soil fertility and reduces the use of nitrogen fertilizers in sustainable agricultural systems. The research aimed to determine the effect in field of the native strain Rhizobium leguminosarum bv. viciae COL6 on growth, N fixation and agricultural yield in common bean cv. Percal. A field trial was carried out to evaluate the parameters of nodulation, biomass, nitrogen fixation, yield components and agricultural yield in common bean. In addition, soil analyzes were performed before and after experimentation. Prior to planting an inoculant was formulated with the strain COL6. The seeds of cv. Percal were mixed in 250 g inoculant per 10 kg of seed and it was sown manually in the soil. The results of growth, N fixation (total N % and agricultural yield demonstrated the positive effect of inoculation with the bacterial inoculant comparable with chemical fertilizers. The increase in agricultural yield with bacterial inoculant application and chemical fertilization with respect to control was 62 and 64 %, respectively. The results of the soil analysis were variable with respect to the initial physical-chemical characteristics. The promising results of bacterial inoculant use and the effective stimulation in common bean, open the doors for its potential use in Ecuadorian soils and with that the desired potential yield can be reached.

  3. Rhizobium leguminosarum bv. viciae 3841 Adapts to 2,4-Dichlorophenoxyacetic Acid with "Auxin-Like" Morphological Changes, Cell Envelope Remodeling and Upregulation of Central Metabolic Pathways.

    Directory of Open Access Journals (Sweden)

    Supriya V Bhat

    Full Text Available There is a growing need to characterize the effects of environmental stressors at the molecular level on model organisms with the ever increasing number and variety of anthropogenic chemical pollutants. The herbicide 2,4-dichlorophenoxyacetic acid (2,4-D, as one of the most widely applied pesticides in the world, is one such example. This herbicide is known to have non-targeted undesirable effects on humans, animals and soil microbes, but specific molecular targets at sublethal levels are unknown. In this study, we have used Rhizobium leguminosarum bv. viciae 3841 (Rlv as a nitrogen fixing, beneficial model soil organism to characterize the effects of 2,4-D. Using metabolomics and advanced microscopy we determined specific target pathways in the Rlv metabolic network and consequent changes to its phenotype, surface ultrastructure, and physical properties during sublethal 2,4-D exposure. Auxin and 2,4-D, its structural analogue, showed common morphological changes in vitro which were similar to bacteroids isolated from plant nodules, implying that these changes are related to bacteroid differentiation required for nitrogen fixation. Rlv showed remarkable adaptation capabilities in response to the herbicide, with changes to integral pathways of cellular metabolism and the potential to assimilate 2,4-D with consequent changes to its physical and structural properties. This study identifies biomarkers of 2,4-D in Rlv and offers valuable insights into the mode-of-action of 2,4-D in soil bacteria.

  4. Revised description and classification of atypical isolates of Pasteurella multocida from bovine lungs based on genotypic characterization to include variants previously classified as biovar 2 of Pasteurella canis and Pasteurella avium

    DEFF Research Database (Denmark)

    Christensen, Henrik; Angen, Øystein; Olsen, John E.

    2004-01-01

    Strains deviating in key phenotypic characters, mainly isolated from cases of bovine pneumonia in five European countries, were genotyped in order to examine their genotypic relationship with Pasteurella multocida. Twenty-two strains of Pasteurella avium biovar 2, including variants in indole, xy...

  5. High spatial variation in population size and symbiotic performance of Rhizobium leguminosarum bv. trifolii with white clover in New Zealand pasture soils

    Science.gov (United States)

    Tillard, Guyléne; van Ham, Robert; Ballard, Ross; Farquharson, Elizabeth; Gerard, Emily; Geurts, Rene; Brown, Matthew; Ridgway, Hayley; O’Callaghan, Maureen

    2018-01-01

    Biological nitrogen fixation through the legume-rhizobia symbiosis is important for sustainable pastoral production. In New Zealand, the most widespread and valuable symbiosis occurs between white clover (Trifolium repens L.) and Rhizobium leguminosarum bv. trifolii (Rlt). As variation in the population size (determined by most probable number assays; MPN) and effectiveness of N-fixation (symbiotic potential; SP) of Rlt in soils may affect white clover performance, the extent in variation in these properties was examined at three different spatial scales: (1) From 26 sites across New Zealand, (2) at farm-wide scale, and (3) within single fields. Overall, Rlt populations ranged from 95 to >1 x 108 per g soil, with variation similar at the three spatial scales assessed. For almost all samples, there was no relationship between rhizobia population size and ability of the population to fix N during legume symbiosis (SP). When compared with the commercial inoculant strain, the SP of soils ranged between 14 to 143% efficacy. The N-fixing ability of rhizobia populations varied more between samples collected from within a single hill country field (0.8 ha) than between 26 samples collected from diverse locations across New Zealand. Correlations between SP and calcium and aluminium content were found in all sites, except within a dairy farm field. Given the general lack of association between SP and MPN, and high spatial variability of SP at single field scale, provision of advice for treating legume seed with rhizobia based on field-average MPN counts needs to be carefully considered. PMID:29489845

  6. Brucellosis seroprevalence in Bali cattle with reproductive failure in South Sulawesi and Brucella abortus biovar 1 genotypes in the Eastern Indonesian archipelago.

    Science.gov (United States)

    Muflihanah, Hanah; Hatta, Mochammad; Rood, Ente; Scheelbeek, Pauline; Abdoel, Theresia H; Smits, Henk L

    2013-11-26

    Brucellosis is a major cause of infertility and reproductive failure in livestock. While cattle in the Eastern Indonesian archipelago suffers from reproductive problems information on bovine brucellosis in the region is fragmentary. The control of brucellosis requires a major and prolonged effort and confirmation of the infection by isolation with detailed knowledge of the spread of the infection is essential when planning a control program. Serological investigation of Brucella infection in beef cattle tended under extensive farming conditions revealed a high seroprevalence (19.3%; 95% CI, 17-22) in the compliment fixation tests. The results of a rapid and simple field test correlated well with the Rose Bengal test (kappa, 0.917) and indicated an acceptable sensitivity (87.5%) and specificity (98.1%) compared with the complement fixation test. Reproductive failure was reported for 39.0% of the cows with a loss of calves due to abortion or early death amounting to 19.3%. Past reproductive failure did not, however, correlate with seropositivity in the complement fixation test (RP = 1.21; P = 0.847). B. abortus biovar 1 was freshly isolated from the hygromas of two cows and together with thirty banked isolates collected since 1990 from different parts of Sulawesi and Timor eight related genotypes could be distinguished with one genotype being identical to that of an isolate (BfR91) from Switzerland. The Indonesian genotypes formed together with BfR91 and one African and one North American isolate a distinct branch on the B. abortus biovar 1 dendogram. Bovine brucellosis appears to be widespread in the Eastern Indonesian archipelago and calls for urgent intervention. The fresh isolation of the pathogen together with the observed high seroprevalence demonstrates the presence and frequent exposure of cattle in the area to the pathogen. The application of a rapid and simple field test for brucellosis could be very useful for the quick screening of cattle at the pen side.

  7. Crystallization and X-ray diffraction analysis of an l-arabinonate dehydratase from Rhizobium leguminosarum bv. trifolii and a d-xylonate dehydratase from Caulobacter crescentus

    Energy Technology Data Exchange (ETDEWEB)

    Rahman, Mohammad Mubinur [University of Eastern Finland, Joensuu Campus, PO Box 111, FIN-80101 Joensuu (Finland); Andberg, Martina; Koivula, Anu [VTT Technical Research Centre of Finland Ltd, PO Box 1000, FIN-02044 VTT Espoo (Finland); Rouvinen, Juha; Hakulinen, Nina, E-mail: nina.hakulinen@uef.fi [University of Eastern Finland, Joensuu Campus, PO Box 111, FIN-80101 Joensuu (Finland)

    2016-07-13

    l-Arabinonate dehydratase and d-xylonate dehydratase from the IlvD/EDD family were crystallized by the vapour-diffusion method. Diffraction data sets were collected to resolutions of 2.40 and 2.66 Å from crystals of l-arabinonate dehydratase and d-xylonate dehydratase, respectively. l-Arabinonate dehydratase (EC 4.2.1.25) and d-xylonate dehydratase (EC 4.2.1.82) are two enzymes that are involved in a nonphosphorylative oxidation pathway of pentose sugars. l-Arabinonate dehydratase converts l-arabinonate into 2-dehydro-3-deoxy-l-arabinonate, and d-xylonate dehydratase catalyzes the dehydration of d-xylonate to 2-dehydro-3-deoxy-d-xylonate. l-Arabinonate and d-xylonate dehydratases belong to the IlvD/EDD family, together with 6-phosphogluconate dehydratases and dihydroxyacid dehydratases. No crystal structure of any l-arabinonate or d-xylonate dehydratase is available in the PDB. In this study, recombinant l-arabinonate dehydratase from Rhizobium leguminosarum bv. trifolii (RlArDHT) and d-xylonate dehydratase from Caulobacter crescentus (CcXyDHT) were heterologously expressed in Escherichia coli and purified by the use of affinity chromatography followed by gel-filtration chromatography. The purified proteins were crystallized using the hanging-drop vapour-diffusion method at 293 K. Crystals of RlArDHT that diffracted to 2.40 Å resolution were obtained using sodium formate as a precipitating agent. They belonged to space group P2{sub 1}, with unit-cell parameters a = 106.07, b = 208.61, c = 147.09 Å, β = 90.43°. Eight RlArDHT molecules (two tetramers) in the asymmetric unit give a V{sub M} value of 3.2 Å{sup 3} Da{sup −1} and a solvent content of 62%. Crystals of CcXyDHT that diffracted to 2.66 Å resolution were obtained using sodium formate and polyethylene glycol 3350. They belonged to space group C2, with unit-cell parameters a = 270.42, b = 236.13, c = 65.17 Å, β = 97.38°. Four CcXyDHT molecules (a tetramer) in the asymmetric unit give a V{sub M

  8. INFLUÊNCIA DA DENSIDADE DE INÓCULO DE Fusarium solani f.sp. phaseoli NA SEVERIDADE DA PODRIDÃO RADICULAR SECA DO FEIJOEIRO EFFECT OF Fusarium solani f.sp. phaseoli INOCULUM DENSITY ON DRY ROOT ROT SEVERITY IN THE COMMON BEAN

    Directory of Open Access Journals (Sweden)

    Gesimária Ribeiro Costa

    2007-09-01

    Full Text Available

    Foram testadas quatro densidades de inóculo de Fusarium solani, em gramas por litro de solo (1,0; 2,0; 4,0 e 8,0 e um tratamento testemunha, em solo tipo Latossolo Vermelho-Escuro, cultivado e não cultivado, com o objetivo de determinar a densidade mínima de inóculo no solo necessária para a ocorrência de podridão radicular seca do feijoeiro. Como variáveis respostas foram avaliadas: número de microorganismos totais do solo, número de propágulos de F. solani, atividade microbiológica total do solo e severidade da doença em plântulas. Os resultados indicaram que a densidade de inóculo do fungo variou com o tipo de solo. Para um solo não cultivado a densidade necessária para causar a doença esteve acima de 5.127 propágulos por grama de solo, enquanto para o solo cultivado a densidade de inóculo para causar doença foi de 3.701 propágulos por grama de solo. Os índices de doença em plântulas cultivadas sob o solo cultivado foram duas vezes superiores ao índice de doença de plântulas sob o solo não cultivado. A atividade microbiológica total nos solos, determinada pela desidrogenase de fluorescina diacetato, não se correlacionou com a população dos microorganismos, indicando que a simples presença desses não implica em que estejam ativos.

    PALAVRAS-CHAVE: Solo supressivo; solo conducivo; Phaseolus vulgaris.

    Four densities of Fusarium solani inoculum (1, 2, 4 and 8 g/L of soil were tested for determining the minimum inoculum density for the occurrence of bean dry root rot, in two soil types. The response variables evaluated were the total number of microorganisms in the soil, the number of F. solani f. sp. phaseoli propagules, total soil microbial activity and seedling disease severity

  9. A Suppressor of the Menadione-Hypersensitive Phenotype of a Xanthomonas campestris pv. phaseoli oxyR Mutant Reveals a Novel Mechanism of Toxicity and the Protective Role of Alkyl Hydroperoxide Reductase

    Science.gov (United States)

    Vattanaviboon, Paiboon; Whangsuk, Wirongrong; Mongkolsuk, Skorn

    2003-01-01

    We isolated menadione-resistant mutants of Xanthomonas campestris pv. phaseoli oxyR (oxyRXp). The oxyRR2Xp mutant was hyperresistant to the superoxide generators menadione and plumbagin and was moderately resistant to H2O2 and tert-butyl hydroperoxide. Analysis of enzymes involved in oxidative-stress protection in the oxyRR2Xp mutant revealed a >10-fold increase in AhpC and AhpF levels, while the levels of superoxide dismutase (SOD), catalase, and the organic hydroperoxide resistance protein (Ohr) were not significantly altered. Inactivation of ahpC in the oxyRR2Xp mutant resulted in increased sensitivity to menadione killing. Moreover, high levels of expression of cloned ahpC and ahpF in the oxyRXp mutant complemented the menadione hypersensitivity phenotype. High levels of other oxidant-scavenging enzymes such as catalase and SOD did not protect the cells from menadione toxicity. These data strongly suggest that the toxicity of superoxide generators could be mediated via organic peroxide production and that alkyl hydroperoxide reductase has an important novel function in the protection against the toxicity of these compounds in X. campestris. PMID:12591894

  10. Sequence diversity in the Dickeya fliC gene: phylogeny of the Dickeya genus and TaqMan® PCR for 'D. solani', new biovar 3 variant on potato in Europe.

    Science.gov (United States)

    Van Vaerenbergh, Johan; Baeyen, Steve; De Vos, Paul; Maes, Martine

    2012-01-01

    Worldwide, Dickeya (formerly Erwinia chrysanthemi) is causing soft rot diseases on a large diversity of crops and ornamental plants. Strains affecting potato are mainly found in D. dadantii, D. dianthicola and D. zeae, which appear to have a marked geographical distribution. Furthermore, a few Dickeya isolates from potato are attributed to D. chrysanthemi and D. dieffenbachiae. In Europe, isolates of Erwinia chrysanthemi biovar 1 and biovar 7 from potato are now classified in D. dianthicola. However, in the past few years, a new Dickeya biovar 3 variant, tentatively named 'Dickeya solani', has emerged as a common major threat, in particular in seed potatoes. Sequences of a fliC gene fragment were used to generate a phylogeny of Dickeya reference strains from culture collections and with this reference backbone, to classify pectinolytic isolates, i.e. Dickeya spp. from potato and ornamental plants. The reference strains of the currently recognized Dickeya species and 'D. solani' were unambiguously delineated in the fliC phylogram. D. dadantii, D. dianthicola and 'D. solani' displayed unbranched clades, while D. chrysanthemi, D. zeae and D. dieffenbachiae branched into subclades and lineages. Moreover, Dickeya isolates from diagnostic samples, in particular biovar 3 isolates from greenhouse ornamentals, formed several new lineages. Most of these isolates were positioned between the clade of 'D. solani' and D. dadantii as transition variants. New lineages also appeared in D. dieffenbachiae and in D. zeae. The strains and isolates of D. dianthicola and 'D. solani' were differentiated by a fliC sequence useful for barcode identification. A fliC TaqMan®real-time PCR was developed for 'D. solani' and the assay was provisionally evaluated in direct analysis of diagnostic potato samples. This molecular tool can support the efforts to control this particular phytopathogen in seed potato certification.

  11. Sequence diversity in the Dickeya fliC gene: phylogeny of the Dickeya genus and TaqMan® PCR for 'D. solani', new biovar 3 variant on potato in Europe.

    Directory of Open Access Journals (Sweden)

    Johan Van Vaerenbergh

    Full Text Available Worldwide, Dickeya (formerly Erwinia chrysanthemi is causing soft rot diseases on a large diversity of crops and ornamental plants. Strains affecting potato are mainly found in D. dadantii, D. dianthicola and D. zeae, which appear to have a marked geographical distribution. Furthermore, a few Dickeya isolates from potato are attributed to D. chrysanthemi and D. dieffenbachiae. In Europe, isolates of Erwinia chrysanthemi biovar 1 and biovar 7 from potato are now classified in D. dianthicola. However, in the past few years, a new Dickeya biovar 3 variant, tentatively named 'Dickeya solani', has emerged as a common major threat, in particular in seed potatoes. Sequences of a fliC gene fragment were used to generate a phylogeny of Dickeya reference strains from culture collections and with this reference backbone, to classify pectinolytic isolates, i.e. Dickeya spp. from potato and ornamental plants. The reference strains of the currently recognized Dickeya species and 'D. solani' were unambiguously delineated in the fliC phylogram. D. dadantii, D. dianthicola and 'D. solani' displayed unbranched clades, while D. chrysanthemi, D. zeae and D. dieffenbachiae branched into subclades and lineages. Moreover, Dickeya isolates from diagnostic samples, in particular biovar 3 isolates from greenhouse ornamentals, formed several new lineages. Most of these isolates were positioned between the clade of 'D. solani' and D. dadantii as transition variants. New lineages also appeared in D. dieffenbachiae and in D. zeae. The strains and isolates of D. dianthicola and 'D. solani' were differentiated by a fliC sequence useful for barcode identification. A fliC TaqMan®real-time PCR was developed for 'D. solani' and the assay was provisionally evaluated in direct analysis of diagnostic potato samples. This molecular tool can support the efforts to control this particular phytopathogen in seed potato certification.

  12. High resolution melting (HRM) analysis as a new tool for rapid identification of Salmonella enterica serovar Gallinarum biovars Pullorum and Gallinarum.

    Science.gov (United States)

    Ren, Xingxing; Fu, Ying; Xu, Chenggang; Feng, Zhou; Li, Miao; Zhang, Lina; Zhang, Jianmin; Liao, Ming

    2017-05-01

    Salmonella enterica serovar Gallinarum biovars Pullorum and Gallinarum represent the most common causative agents of chicken salmonellosis, which result in high mortality and morbidity throughout the world. It is difficult and laborious to discriminate these diseases based on biochemical or phenotypic methods. Herein, we report the development of a single nucleotide polymorphism (SNP) PCR-high resolution melt (PCR-HRM) assay for the detection and discrimination of both S. Pullorum and S. Gallinarun. The gene rfbS, which encodes a factor involved in the biosynthesis of ADP paratose in serogroup D of Salmonella, has been identified as a robust genetic marker for the identification of S. Pullorum and S. Gallinarun based on polymorphisms at positions 237 and 598. Therefore, PCR-HRM analyses were used to characterize this gene. A total of 15 reference and 33 clinical isolates of Salmonella and related Gram-negative bacteria were detected using 2 sets of primers. Our PCR-HRM assay could distinguish S. Pullorum from S. Gallinarun and other strains using the primer pair SP-237F/237R. Similarly, S. Gallinarun could be distinguished from S. Pullorum and other strains using primer set SG-598F/598R. These 2 assays showed high specificity (100%) for both S. Pullorum and S. Gallinarun; the sensitivity of these 2 assays was at least 100-fold greater than that of the allele-specific PCR assay. This present study demonstrated that HRM analysis represents a potent, simple, and economic tool for the rapid, specific, and sensitive detection of S. Pullorum and S. Gallinarun. Our approach also may aid efforts for purification of Avian Salmonella disease. © 2016 Poultry Science Association Inc.

  13. Plasmids of Raw Milk Cheese Isolate Lactococcus lactis subsp. lactis Biovar diacetylactis DPC3901 Suggest a Plant-Based Origin for the Strain ▿ †

    Science.gov (United States)

    Fallico, Vincenzo; McAuliffe, Olivia; Fitzgerald, Gerald F.; Ross, R. Paul

    2011-01-01

    The four-plasmid complement of the raw milk cheese isolate Lactococcus lactis subsp. lactis biovar diacetylactis DPC3901 was sequenced, and some genetic features were functionally analyzed. The complete sequences of pVF18 (18,977 bp), pVF21 (21,739 bp), pVF22 (22,166 bp), and pVF50 (53,876 bp) were obtained. Each plasmid contained genes not previously described for Lactococcus, in addition to genes associated with plant-derived lactococcal strains. Most of the novel genes were found on pVF18 and encoded functions typical of bacteria associated with plants, such as activities of plant cell wall modification (orf11 and orf25). In addition, a predicted high-affinity regulated system for the uptake of cobalt was identified (orf19 to orf21 [orf19-21]), which has a single database homolog on a plant-derived Leuconostoc plasmid and whose functionality was demonstrated following curing of pVF18. pVF21 and pVF22 encode additional metal transporters, which, along with orf19-21 of pVF18, could enhance host ability to uptake growth-limiting amounts of biologically essential ions within the soil. In addition, vast regions from pVF50 and pVF21 share significant homology with the plant-derived lactococcal plasmid pGdh442, which is indicative of extensive horizontal gene transfer and recombination between these plasmids and suggests a common plant niche for their hosts. Phenotypes associated with these regions include glutamate dehydrogenase activity and Na+ and K+ transport. The presence of numerous plant-associated markers in L. lactis DPC3901 suggests a plant origin for the raw milk cheese isolate and provides for the first time the genetic basis to support the concept of the plant-milk transition for Lactococcus strains. PMID:21803914

  14. Licheniocin 50.2 and Bacteriocins from Lactococcus lactis subsp. lactis biovar. diacetylactis BGBU1-4 Inhibit Biofilms of Coagulase Negative Staphylococci and Listeria monocytogenes Clinical Isolates.

    Science.gov (United States)

    Cirkovic, Ivana; Bozic, Dragana D; Draganic, Veselin; Lozo, Jelena; Beric, Tanja; Kojic, Milan; Arsic, Biljana; Garalejic, Eliana; Djukic, Slobodanka; Stankovic, Slavisa

    2016-01-01

    Coagulase negative staphylococci (CoNS) and Listeria monocytogenes have important roles in pathogenesis of various genital tract infections and fatal foetomaternal infections, respectively. The aim of our study was to investigate the inhibitory effects of two novel bacteriocins on biofilms of CoNS and L. monocytogenes genital isolates. The effects of licheniocin 50.2 from Bacillus licheniformis VPS50.2 and crude extract of bacteriocins produced by Lactococcus lactis subsp. lactis biovar. diacetylactis BGBU1-4 (BGBU1-4 crude extract) were evaluated on biofilm formation and formed biofilms of eight CoNS (four S. epidermidis, two S. hominis, one S. lugdunensis and one S. haemolyticus) and 12 L. monocytogenes genital isolates. Licheniocin 50.2 and BGBU1-4 crude extract inhibited the growth of both CoNS and L. monocytogenes isolates, with MIC values in the range between 200-400 AU/ml for licheniocin 50.2 and 400-3200 AU/ml for BGBU1-4 crude extract. Subinhibitory concentrations (1/2 × and 1/4 × MIC) of licheniocin 50.2 inhibited biofilm formation by all CoNS isolates (p < 0.05, respectively), while BGBU1-4 crude extract inhibited biofilm formation by all L. monocytogenes isolates (p < 0.01 and p < 0.05, respectively). Both bacteriocins in concentrations of 100 AU/mL and 200 AU/mL reduced the amount of 24 h old CoNS and L. monocytogenes biofilms (p < 0.05, p < 0.01, p < 0.001). This study suggests that novel bacteriocins have potential to be used for genital application, to prevent biofilm formation and/or to eradicate formed biofilms, and consequently reduce genital and neonatal infections by CoNS and L. monocytogenes.

  15. The Fast-Growing Brucella suis Biovar 5 Depends on Phosphoenolpyruvate Carboxykinase and Pyruvate Phosphate Dikinase but Not on Fbp and GlpX Fructose-1,6-Bisphosphatases or Isocitrate Lyase for Full Virulence in Laboratory Models

    Directory of Open Access Journals (Sweden)

    Amaia Zúñiga-Ripa

    2018-04-01

    Full Text Available Bacteria of the genus Brucella infect a range of vertebrates causing a worldwide extended zoonosis. The best-characterized brucellae infect domestic livestock, behaving as stealthy facultative intracellular parasites. This stealthiness depends on envelope molecules with reduced pathogen-associated molecular patterns, as revealed by the low lethality and ability to persist in mice of these bacteria. Infected cells are often engorged with brucellae without signs of distress, suggesting that stealthiness could also reflect an adaptation of the parasite metabolism to use local nutrients without harming the cell. To investigate this, we compared key metabolic abilities of Brucella abortus 2308 Wisconsin (2308W, a cattle biovar 1 virulent strain, and B. suis 513, the reference strain of the ancestral biovar 5 found in wild rodents. B. suis 513 used a larger number of C substrates and showed faster growth rates in vitro, two features similar to those of B. microti, a species phylogenomically close to B. suis biovar 5 that infects voles. However, whereas B. microti shows enhanced lethality and reduced persistence in mice, B. suis 513 was similar to B. abortus 2308W in this regard. Mutant analyses showed that B. suis 513 and B. abortus 2308W were similar in that both depend on phosphoenolpyruvate synthesis for virulence but not on the classical gluconeogenic fructose-1,6-bisphosphatases Fbp-GlpX or on isocitrate lyase (AceA. However, B. suis 513 used pyruvate phosphate dikinase (PpdK and phosphoenolpyruvate carboxykinase (PckA for phosphoenolpyruvate synthesis in vitro while B. abortus 2308W used only PpdK. Moreover, whereas PpdK dysfunction causes attenuation of B. abortus 2308W in mice, in B. suis, 513 attenuation occurred only in the double PckA-PpdK mutant. Also contrary to what occurs in B. abortus 2308, a B. suis 513 malic enzyme (Mae mutant was not attenuated, and this independence of Mae and the role of PpdK was confirmed by the lack of

  16. Structures of two cell wall-associated polysaccharides of a Streptococcus mitis biovar 1 strain. A unique teichoic acid-like polysaccharide and the group O antigen which is a C-polysaccharide in common with pneumococci

    DEFF Research Database (Denmark)

    Bergström, N; Jansson, P.-E.; Kilian, Mogens

    2000-01-01

    The cell wall of Streptococcus mitis biovar 1 strain SK137 contains the C-polysaccharide known as the common antigen of a closely related species Streptococcus pneumoniae, and a teichoic acid-like polysaccharide with a unique structure. The two polysaccharides are different entities and could...... be partially separated by gel chromatography. The structures of the two polysaccharides were determined by chemical methods and by NMR spectroscopy. The teichoic acid-like polymer has a heptasaccharide phosphate repeating unit with the following structure: The structure neither contains ribitol nor glycerol...... phosphate as classical teichoic acids do, thus we have used the expression teichoic acid-like for this polysaccharide. The following structure of the C-polysaccharide repeating unit was established: where AAT is 2-acetamido-4-amino-2,4, 6-trideoxy-D-galactose. It has a carbohydrate backbone identical...

  17. Number and Effectiveness of Pea Rhizobia in Danish Soils

    DEFF Research Database (Denmark)

    Engvild, K.C.

    1989-01-01

    Most of 44 Danish soils tested contain between 1000 and 10 000 pea rhizobia (Rhizobium leguminosarum biovar viceae) per gram. Pea rhizobia were not detected in acid moor and forest soils. Only one case of failed nodulation in peas in the field has been noted, in spots in a reclaimed sandy heath m...

  18. 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

  19. Diversity of Rhizobium-Phaseolus vulgaris symbiosis: Overview and perspectives

    International Nuclear Information System (INIS)

    Martinez-Romero, Esperanza

    2001-01-01

    Common bean (Phaseolus vulgaris) has become a cosmopolitan crop, but was originally domesticated in the Americas and has been grown in Latin America for several thousand years. Consequently an enormous diversity of bean nodulating bacteria have developed and in the centers of origin the predominant species in bean nodules is R. etli. In some areas of Latin America, inoculation, which normally promotes nodulation and nitrogen fixation is hampered by the prevalence of native strains. Many other species in addition to R. etli have been found in bean nodules in regions where bean has been introduced. Some of these species such as R. leguminosarum bv. phaseoli, R. gallicum bv. phaseoli and R. giardinii bv. phaseoli might have arisen by acquiring the phaseoli plasmid from R. etli. Others, like R. trap id, are well adapted to acid soils and high temperatures and are good inoculants for bean under these conditions. The large number of rhizobia species capable of nodulating bean supports that bean is a promiscuous host and a diversity of bean-rhizobia interactions exists. Large ranges of dinitrogen fixing capabilities have been documented among bean cultivars and commercial beans have the lowest values among legume crops. Knowledge on bean symbiosis is still incipient but could help to improve bean biological nitrogen fixation. (author)

  20. Effect of neem cake/fertilizers on symbiotic and non-symbiotic N2 fixing bacteria

    International Nuclear Information System (INIS)

    Akhtar, S.; Solangi, A.H.; Gilani, G.; Pirzada, M.H.

    2002-01-01

    Neem cake amendment in soil at 1.3% no adverse effect on the population of four symbiotic Rhizobium species viz., japonicum, R. leguminosarum, R. Phaseoli and R. Fredii and three non-symbiotic free living nitrogen fixers bacteria viz., Pseudomonas diazotrophicus, Klebsiella planticola and Enterobacter cloacae. Neem cake extracted with n-hexane stimulated the growth of Rhizobium species in vitro, whereas Neem cake expeller extracted neither inhibited nor stimulated the growth of Rhizobium species except for R. Fredii, whose was slightly retarded. The fertilizers (urea, NPK and DAP) had no adverse effect on these bacteria even at the dosage ten times higher the recommended dose. (author)

  1. Effects of Rhizobuim leguminosarum inoculation on the growth and ...

    African Journals Online (AJOL)

    g per plant in 2008 and 2009 seasons, respectively. The findings suggested that inoculation of Mucuna flagellipies with Rhizobium is beneficial and produced high seed yield and could be use as biofertilizer an alternative to nitrogen fertilizer.

  2. Occurrence of plant growth-promoting traits in clover-nodulating rhizobia strains isolated from different soils in Rio Grande do Sul state Ocorrência de características de promoção de crescimento em rizóbios noduladores de trevo isolados de solos do Rio Grande do Sul

    Directory of Open Access Journals (Sweden)

    Luciano Kayser Vargas

    2009-10-01

    Full Text Available In the last decades, the use of plant growth-promoting rhizobacteria has become an alternative to improve crop production. Rhizobium leguminosarum biovar trifolii is one of the most promising rhizobacteria and is even used with non-legume plants. This study investigated in vitro the occurrence of plant growth-promoting characteristics in several indigenous R. leguminosarum biovar trifolii isolated from soils in the State of Rio Grande do Sul, Brazil. Isolates were obtained at 11 locations and evaluated for indoleacetic acid and siderophore production and inorganic phosphate solubilization. Ten isolates were also molecularly characterized and tested for antagonism against a phytopathogenic fungus and for plant growth promotion of rice seedlings. Of a total of 252 isolates, 59 produced indoleacetic acid, 20 produced siderophores and 107 solubilized phosphate. Some degree of antagonism against Verticillium sp. was observed in all tested isolates, reducing mycelial growth in culture broth. Isolate AGR-3 stood out for increasing root length of rice seedlings, while isolate ELD-18, besides increasing root length in comparison to the uninoculated control, also increased the germination speed index, shoot length, and seedling dry weight. These results confirm the potential of some strains of R. leguminosarum biovar trifolii as plant growth-promoting rhizobacteria.Nas últimas décadas, o uso de rizobactérias promotoras de crescimento de plantas tem se mostrado uma alternativa para aumentar a produção vegetal. Rhizobium leguminosarum biovar trifolii está entre as mais promissoras rizobactérias e tem sido usada também em não leguminosas. O presente trabalho analisou in vitro a ocorrência de características de promoção de crescimento vegetal em isolados de R. leguminosarum biovar trifolii nativos de solos do Rio Grande do Sul. Os isolados foram obtidos de 11 localidades e avaliados quanto à produção de ácido indolacético, produção de

  3. Diversity and numbers of root-nodule bacteria (rhizobia in Polish soils

    Directory of Open Access Journals (Sweden)

    Stefan Martyniuk

    2011-01-01

    Full Text Available Using a sand pouch-plant infection method, populations of several species of root-nodule bacteria (rhizobia were enumerated in eighty soils collected throughout Poland. Rhizobium leguminosarum bv. viciae (symbionts of pea, faba bean, vetch and R. leguminosarum bv. trifolii (symbionts of clover were detected in 77 and 76 soils, respectively. Most of these soils contained moderate and high numbers of these species of the rhizobia. Symbionts of beans, R. leguminosarum bv. phaseoli, were assessed in 76 soils; of this number 15 soils had no detectable populations of bean rhizobia and in 40 soils high or moderate numbers of these bacteria were found. Bradyrhizobium sp. (Lupinus, root-nodule bacteria of lupine and serradella, were absent in 19 soils, out of 80 tested, and 34 soils were colonised by high or moderate populations of bradyrhizobia. Sinorhizobium meliloti, rhizobia nodulating alfalfa, were sparse in the examined soils; with 56 soil containing no detectable numbers of S. meliloti and only 6 soils harbouring high or moderate populations of this species. The estimated numbers of the rhizobia in the studied soils were also related to some physical and chemical properties of these soils.

  4. Effects of high temperature on survival, symbiotic performance and genomic modifications of bean nodulating Rhizobium strains Sobrevivência, fixação de nitrogênio e modificações genéticas em estirpes de Rhizobium sp. efetivas na nodulação do feijoeiro, expostas à altas temperaturas

    Directory of Open Access Journals (Sweden)

    Patrícia P. Pinto

    1998-10-01

    Full Text Available High temperatures can affect the survival, establishment and symbiotic properties of Rhizobium strains. Bean nodulating Rhizobium strains are considered particularly sensitive because on this strains genetic recombinations and/or deletions occur frequently, thus compromising the use of these bacteria as inoculants. In this study R. tropici and R. leguminosarum bv. phaseoli strains isolated from Cerrado soils were exposed to thermal stress and the strains’ growth, survival and symbiotic relationships as well as alterations in their genotypic and phenotypic characteristics were analyzed. After successive thermal shocks at 45ºC for four hours, survival capacity appeared to be strain-specific, independent of thermo-tolerance and was more apparent in R. tropici strains. Certain R. leguminosarum bv. phaseoli strains had significant alterations in plant dry weight and DNA patterns obtained by AP-PCR method. R. tropici strains (with the exception of FJ2.21 were more stable than R. leguminosarum bv. phaseoli strains because no significant phenotypic alterations were observed following thermal treatments and they maintained their original genotypic pattern after inoculation in plants.Altas temperaturas podem afetar a sobrevivência, estabelecimento e as propriedades simbióticas em estirpes de Rhizobium. As estirpes capazes de nodular o feijoeiro têm sido consideradas particularmente sensíveis, porque nessas estirpes é comum a ocorrência de recombinações e/ou deleções genômicas comprometendo, muitas vezes, a sua utilização como inoculantes. Neste trabalho, procurou-se avaliar a capacidade de crescimento e sobrevivência em temperaturas elevadas de estirpes de Rhizobium efetivas na fixação de nitrogênio no feijoeiro isoladas dos cerrados, bem como avaliar suas características fenotípicas e genotípicas após choque térmico. A capacidade de sobrevivência à temperaturas elevadas, avaliada após choques térmicos sucessivos (45ºC por 4

  5. Analysis of Kenyan isolates of Fusarium solani f. sp. phaseoli from ...

    African Journals Online (AJOL)

    All the isolates showed high variability in aerial mycelial growth, mycelia texture, pigmentation (mycelia colour) when cultured on potato dextrose agar medium, and conidial measurements on Spezieller Nahrstoffarmer agar medium. Colonies were grouped into luxuriant, moderately luxuriant and scanty on aerial mycelial ...

  6. Analysis of Kenyan isolates of Fusarium solani f. sp. phaseoli from ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-06-03

    Jun 3, 2008 ... ... reported worldwide, including Mexico, Brazil, Colombia, Peru, Ecuador, Chile,. Venezuela, Costa Rica and Malawi (Kraft et al., 1981). ... growth, mycelial texture and pigments (colour) produced on the mycelia. The nature of ...

  7. Biosynthesis of compatible solutes in rhizobial strains isolated from Phaseolus vulgaris nodules in Tunisian fields

    Directory of Open Access Journals (Sweden)

    Nieto Joaquín J

    2010-07-01

    Full Text Available Abstract Background Associated with appropriate crop and soil management, inoculation of legumes with microbial biofertilizers can improve food legume yield and soil fertility and reduce pollution by inorganic fertilizers. Rhizospheric bacteria are subjected to osmotic stress imposed by drought and/or NaCl, two abiotic constraints frequently found in semi-arid lands. Osmostress response in bacteria involves the accumulation of small organic compounds called compatible solutes. Whereas most studies on rhizobial osmoadaptation have focussed on the model species Sinorhizobium meliloti, little is known on the osmoadaptive mechanisms used by native rhizobia, which are good sources of inoculants. In this work, we investigated the synthesis and accumulations of compatible solutes by four rhizobial strains isolated from root nodules of Phaseolus vulgaris in Tunisia, as well as by the reference strain Rhizobium tropici CIAT 899T. Results The most NaCl-tolerant strain was A. tumefaciens 10c2, followed (in decreasing order by R. tropici CIAT 899, R. leguminosarum bv. phaseoli 31c3, R. etli 12a3 and R. gallicum bv. phaseoli 8a3. 13C- and 1H-NMR analyses showed that all Rhizobium strains synthesized trehalose whereas A. tumefaciens 10c2 synthesized mannosucrose. Glutamate synthesis was also observed in R. tropici CIAT 899, R. leguminosarum bv. phaseoli 31c3 and A. tumefaciens 10c2. When added as a carbon source, mannitol was also accumulated by all strains. Accumulation of trehalose in R. tropici CIAT 899 and of mannosucrose in A. tumefaciens 10c2 was osmoregulated, suggesting their involvement in osmotolerance. The phylogenetic analysis of the otsA gene, encoding the trehalose-6-phosphate synthase, suggested the existence of lateral transfer events. In vivo 13C labeling experiments together with genomic analysis led us to propose the uptake and conversion pathways of different carbon sources into trehalose. Collaterally, the β-1,2-cyclic glucan from R

  8. Multivariate analysis and determination of the best indirect selection criteria to genetic improvement the biological nitrogen fixation ability in common bean genotypes (Phaseolus vulgaris L.

    Directory of Open Access Journals (Sweden)

    Golparvar Reza Ahmad

    2012-01-01

    Full Text Available In order to determine the best indirect selection criteria for genetic improvement of biological nitrogen fixation, sixty four common bean genotypes were cultivated in two randomized complete block design. Genotypes were inoculated with bacteria Rhizobium legominosarum biovar Phaseoli isolate L-109 only in one of the experiments. The second experiment was considered as check for the first. Correlation analysis showed positive and highly significant correlation of majority of the traits with percent of nitrogen fixation. Step-wise regression designated that traits percent of total nitrogen of shoot, number of nodule per plant and biological yield accounted for 92.3 percent of variation exist in percent of nitrogen fixation. Path analysis indicated that these traits have direct and positive effect on percent of nitrogen fixation. Hence, these traits are promising indirect selection criteria for genetic improvement of nitrogen fixation capability in common bean genotypes especially in early generations.

  9. Transformation of pWWO in Rhizobium leguminosarum DPT to Engineer Toluene Degrading Ability for Rhizoremediation

    OpenAIRE

    Goel, Garima; Pandey, Piyush; Sood, Anchal; Bisht, Sandeep; Maheshwari, D. K.; Sharma, G. D.

    2011-01-01

    Rhizoremediation of organic xenobiotics is based on interactions between plants and their associated micro-organisms. The present work was designed to engineer a bacterial system having toluene degradation ability along with plant growth promoting characteristics for effective rhizoremediation. pWWO harboring the genes responsible for toluene breakdown was isolated from Pseudomonas putida MTCC 979 and successfully transformed in Rhizobium DPT. This resulted in a bacterial strain (DPTT) which ...

  10. The symbiosis between Rhizobium leguminosarum and Pisum sativum : regulation of the nitrogenase activity

    NARCIS (Netherlands)

    Appels, M.A.

    1989-01-01

    Bacteria of the genus Rhizobium can form a symbiosis with plants of the family Leguminosae. Both bacteria and plant show considerable biochemical and morphological changes in order to develop and carry out the symbiosis. The Rhizobia

  11. Análisis funcional de DsbA una tiol oxidorreductasa de Rhizobium leguminosarum

    OpenAIRE

    Vega de los Reyes, Rosa Mª

    2009-01-01

    Las proteínas que se exportan fuera del citoplasma bacteriano constituyen hasta un 20% del total de proteínas sintetizadas. Algunas de estas proteínas necesitan adquirir su conformación tridimensional particular en el citoplasma, incorporando frecuentemente cofactores metálicos antes de su transporte. Dicho transporte está mediado por el sistema Tat que es capaz de transportar proteínas que presentan un péptido señal característico en conformación plegada. Se ha postulado DsbA como proteína d...

  12. Identification and characterization of symbiotic genes on the Rhizobium leguminosarum pre sym-plasmid

    NARCIS (Netherlands)

    Schetgens, T.M.P.

    1986-01-01

    Bacteria of the genera Rhizobium and Bradyrhizobium are unique in their quality to form nitrogen-fixing root nodules in symbiosis with leguminous plants. In fast-growing Rhizobium bacteria the genes involved in host recognition and nodule

  13. Comparative effectiveness of different Rhizobium sp. for improving growth and yield of maize (Zea mays L.

    Directory of Open Access Journals (Sweden)

    Ijaz Mehboob, Zahir Ahmad Zahir, Muhammad Arshad, Muhammad Khalid

    2012-05-01

    Full Text Available During the last couple of decades, it has been demonstrated that rhizobia can associate with roots of non-legumes also without forming true nodules, and can promote their growth by using one or more of the direct or indirect mechanisms of actions. This work examines the growth and yield responses of maize to inoculation with different species of rhizobia, isolated from the root nodules of chickpea (Cicer arietinum L., lentil (Lens culinaris M. and mung bean (Vigna radiata L. in pots and fields. Twenty isolates of rhizobia were isolated from root nodules each of mung bean, lentil and chickpea and were screened under axenic conditions. On the basis of their promising performance under axenic conditions, nine most efficient isolates (three from each legume host were selected, characterized and further evaluated for their growth promoting activities by conducting pot and field experiments. Results of pot experiment revealed that maximum increase in grain yield, 1000 grain weight, N, P and K uptake (up to 47.89, 54.52, 73.46, 84.66 and 59.19% by CRI28, respectively, over un-inoculated control was produced by the isolate of Mesorhizobium ciceri. Whereas, maximum improvement in rest of the parameters was caused by the isolates of Rhizobium phaseoli (i.e. fresh biomass, straw yield and root length up to 36.30% by A18, 25.46% by S6 and 81.89% by A18, respectively over un-inoculated control. Rhizobium leguminosarum isolates came out to be the least effective among the species tested. Similarly, all the selected isolates improved the growth and yield attributing parameters in fields as well but with varying capacity compared with un-inoculated control. The selected isolates of Mesorhizobium ciceri and Rhizobium phaseoli again remained superior compared to the isolates of Rhizobium leguminosarum under field conditions. The results of this study imply that rhizobium species had potential to promote growth and yield of maize but this technology should be

  14. Experimental transmission of Corynebacterium pseudotuberculosis biovar equi in horses by house flies

    Science.gov (United States)

    The route of Corynebacterium pseudotuberculosis infection in horses remains undetermined, but transmission by insects is suspected. Scientists from CMAVE and Auburn University investigated house flies (Musca domestica L.) as possible vectors. Three ponies were directly inoculated with C. pseudotuber...

  15. Re-examination of cellular cyclic beta-1,2-glucans of Rhizobiaceae: distribution of ring sizes and degrees of glycerol-1-phosphate substitution.

    Science.gov (United States)

    Zevenhuizen, L P; van Veldhuizen, A; Fokkens, R H

    1990-04-01

    Gel-filtration and thin layer chromatography of low molecular weight carbohydrates from culture filtrates of Agrobacterium radiobacter, Isolate II, have shown, that next to the neutral beta-1,2-glucan fraction a major acidic fraction was present which was found to be glycerophosphorylated cyclic beta-1,2-glucans. Re-examination of cyclic beta-1,2-glucan preparations which had been obtained by extraction of Rhizobium cells with hot phenol-water also showed these acidic modified beta-1,2-glucans to be present. Cyclic beta-1,2-glucans from R. leguminosarum (9 strains) and of R. phaseoli (1 strain) had ring size distribution with degrees of polymerisation (DPs) of 19 and 20 as major ring sizes of which a minor part was glycerophosphorylated; beta-1,2-glucans of R. trifolii (3 strains) had ring sizes with DPs measuring 19-22 as prominent components which were largely unsubstituted, and R. meliloti (7 strains) had beta-1,2-glucans with ring size distributions extending to still higher DPs of 19-25 of which the major part appeared to be glycerophosphorylated.

  16. Biofertilisasi bakteri rhizobium pada tanaman kedelai (Glycine max (L MERR.

    Directory of Open Access Journals (Sweden)

    Tini Surtiningsih

    2012-02-01

    Full Text Available The aim of this research want to know the influence of the addition Rhizobium bacteria species, dose and combination both ofthem, on growth and production of soybean plant (Glycine max (L Merr.. The experimental design of this research was factorial design4×2, 4 species of Rhizobium are R1 = Rhizobium japonicum, R2 = R. phaseoli, R3 = R. leguminosarum, R4 = mixture of R1, R2 andR3, and 2 dose of inoculan Rhizobium (D1 = 5 m/plant, and D2 = 10 ml/plant with 1010 sel bacteria/ml and 5 replications. Independentvariable is species of Rhizobium, dose of inoculan Rhizobium and combination both of them. Dependent variable is dry matter, weightof nodules and dry weight of seeds. The harvest data was analyzed by Kruskal-Wallis Test using 5% level (a = 0.05 followed by Mann-Whitney Test. The result of this research show that species of Rhizobium, dose of inoculan Rhizobium and combination both of thempresent insignificant result (a > 0.05 on soybean growth and production, but the mixture of Rhizobium species with high level doseof bacteria, present better result than single species with low dose of bacteria.

  17. Extracellular polysaccharides are involved in the attachment of Azospirillum brasilense and Rhizobium leguminosarum to arbuscular mycorrhizal structures

    Directory of Open Access Journals (Sweden)

    V Bianciotto

    2009-12-01

    Full Text Available Arbuscular mycorrhizal (AM fungi, one of the most important component of the soil microbial community, establish physical interactions with naturally occurring and genetically modified bacterial biofertilizers and biopesticides, commonly referred to as plant growth-promoting rhizobacteria (PGPR. We have used a genetic approach to investigate the bacterial components possibly involved in the attachment of two PGPR (Azospirillum and Rhizobium to AM roots and AM fungal structures. Mutants affected in extracellular polysaccharides (EPS have been tested in in vitro adhesion assays and shown to be strongly impaired in the attachment to both types of surfaces as well as to quartz fibers. Anchoring of rhizobacteria to AM fungal structures may have special ecological and biotechnological significance because it may facilitate colonisation of new rhizospheres by the bacteria, and may be an essential trait for the development of mixed inocula.

  18. Induction of fungal disease resistance in Vicia faba by dual inoculation with Rhizobium leguminosarum and vesicular-arbuscular mycorrhizal fungi.

    Science.gov (United States)

    Rabie, G H

    1998-01-01

    Infection of Vicia faba with Bothytis fabae causes significant decreases in growth vigour, total nitrogen content, number of nodules and nutrient accumulation. Na-uptake and phenolics concentration increased compared to that of noninfected plants. In contrast, dual inoculation of Rhizobium and VA mycorrhizae increased all above parameters suggesting a distinct improvement of the plants. The results also revealed that an inverse correlation may exist between phenolic, calcium, magnesium and zinc concentrations in mycorrhizal plant tissues grown in presence of rhizobial bacteria and the disease severity. From these findings we conclude a possible role of both VA mycorrhizal fungi and rhizobial bacteria in the decrease of susceptibility of plants.

  19. Evaluations and modifications of semi-selective media for improved isolation of Agrobacterium tumefaciens biovar 1 from cultivated walnut

    Science.gov (United States)

    Agrobacterium tumefaciens, the causal agent of crown gall of walnut, is an aerobic, Gram negative bacterium belonging to the family Rhizobiaceae. Like many in this group, A. tumefaciens is a common inhabitant of soil and plant host tissue. Isolation from these complex environments is difficult even ...

  20. Biochemical and genetical analysis reveal a new clade of biovar 3 Dickeya spp. strains isolated from potato in Europe

    NARCIS (Netherlands)

    Slawiak, M.; Beckhoven, van J.R.C.M.; Speksnijder, A.G.C.L.; Czajkowski, R.L.; Grabe, G.; Wolf, van der J.M.

    2009-01-01

    Sixty-five potato strains of the soft rot-causing plant pathogenic bacterium Dickeya spp., and two strains from hyacinth, were characterised using biochemical assays, REP-PCR genomic finger printing, 16S rDNA and dnaX sequence analysis. These methods were compared with nineteen strains representing

  1. Systemic colonization of potato plants by a soil-borne, GFP-tagged strain of Dickeya sp. Biovar 3

    NARCIS (Netherlands)

    Czajkowski, R.L.; Boer, de W.; Velvis, H.; Wolf, van der J.M.

    2010-01-01

    Colonization of potato plants by soilborne, green fluorescent protein (GFP)-tagged Dickeya sp. IPO2254 was investigated by selective plating, epifluorescence stereo microscopy (ESM), and confocal laser scanning microscopy (CLSM). Replicated experiments were carried out in a greenhouse using plants

  2. First report of latent infection of Cyperus rotundus caused by a biovar 3 Dickeya sp. (Syn. Erwinia chrysanthemi) in Israel

    NARCIS (Netherlands)

    Tsror, L.; Lebiush, S.; Erlich, O.; Ben-Daniel, B.; Wolf, van der J.M.

    2010-01-01

    Recent outbreaks of potato blackleg in Israel, caused by Dickeya spp., on plants grown from seed tubers imported from Northern Europe, are of a great concern. The warm climatic conditions during the growing season favour disease expression, and may result in the establishment of the pathogen in the

  3. Immune Responses and Protection against Experimental Brucella suis biovar 1 Challenge in Non-vaccinated or RB51-Vaccinated Cattle

    Science.gov (United States)

    Twenty Hereford heifers, approximately 9 months of age, were vaccinated with saline (control) or 2 x 10**10 CFU of Brucella abortus strain RB51 (RB51) vaccine. Immunologic responses after inoculation demonstrated significantly greater (P<0.05) antibody and proliferative responses to RB51 antigens i...

  4. Effects of ecological factors on the survival and physiology of Ralstonia solanacearum biovar 2 in agricultural drainage water

    NARCIS (Netherlands)

    Elsas, van J.D.; Kastelein, P.; Vries, de P.M.; Overbeek, van L.S.

    2001-01-01

    The fate of Ralstonia solanacearum bv. 2, the causative agent of brown rot in potato, in aquatic habitats of temperate climate regions is still poorly understood. In this study, the population dynamics and the physiological response of R. solanacearum bv. 2 were tested in sterile pure water and in

  5. Effectiveness and competing capacity of native Rhizobium strains evaluated in IX Region soils

    International Nuclear Information System (INIS)

    Barrientos D, Leticia; Mendez A, Edith; Pino N, Ines

    1995-01-01

    Symbiotic nitrogen fixation in legumes reaches its optimum when the host plants are nodulated by highly competitive and effective Rhizobium strains. With the purpose of assessing competition and nitrogen fixing capacity of native Rhizobium leguminosarum bio-var trifolii strains, a greenhouse test was carried out using white clover (Trifolium re-pens) and four kinds of soils, which represent the main agroecological areas of the IX Region. Eleven strains were evaluated, they were both native and collected and were streptomycin sulphate antibiotic resistant. A nitrogen and a nitrogen-less without inoculation testers were used as controls. All pots received a solution of ammonium sulphate marked with 10% 15 N a.e, equivalent to 10 kg ha -1 of N. Rye-grass was used as reference crop, cv. Nu-i. In general, the evaluated strains were very efficient. After three or four cuts they became the only source of nitrogen for the plants. They were also very competitive, getting to over 70% of root infection with regard to those present in soils. In Curacautin and Tolten soils, biological nitrogen fixation begins later than other soils evaluated, which is coincident with soils having a higher nitrogen content. Symbiosis occurs when the soil nitrogen content exhausts or diminishes. (author)

  6. Improvement of Canola (Brassica napus L.) Inoculated with Rhizobium, Azospirillum and/or Mycorrhizal Fungi Under Salinity Stress

    International Nuclear Information System (INIS)

    El-Ghandour, I. A.; Galal, Y.G; Ebraheem, Rabab M.M.; Yousef, Khayria A.

    2004-01-01

    Bio fertilization technology was applied for improving canola plant growth and nutrient acquisition in sandy saline soil ,as a biological mean used to develop plant growth and nutrient uptake under salinity stress. Canola was cultivated in pots packed at rate of 7 kg saline sandy soil pot -1 , and inoculated with Rhizobium leguminosarum biovar viceae, Azospirillum brasilense strain no. 40 and arbuscular mycorrhizal fungi either solely or in combinations of them. Nitrogen fertilizer was added in form ( 15 NH 4 ) 2 SO 4 with 5% 15 N atom excess at rate of 0.99 g N pot -1 . Maximum dry matter accumulation induced by composite inoculation (Rh + Azo + AM). Na concentrations were frequently affected by Rhizobium and /or mycorrhizae while K was affected by Azospirillum and /or mycorrhizae. Azospirillum enhanced Ca uptake whereas Mg content was responded well to composite inoculants of Rh + Azo + AM. Dual inoculation with Rh + Azo resulted in the highest values of N uptake by plant. Similar effect was noticed with P uptake when dual inoculums of Azo + AM were applied. Data of 15 N isotope showed that the highest portion and value of N 2 -fixed was recorded with composite inoculums followed by dual inoculations. On the other hand, the infection with AM fungi gave the highest proportion of N derived from fertilizer as compared to other inoculants or uninoculated control. In the same trend, the fertilizer use efficiency (FUE%) was enhanced by AM infection. Dual inoculums of Rh + Azo induced highest content of proline in leaves. (Authors)

  7. A common genomic framework for a diverse assembly of plasmids in the symbiotic nitrogen fixing bacteria.

    Directory of Open Access Journals (Sweden)

    Lisa C Crossman

    2008-07-01

    Full Text Available This work centres on the genomic comparisons of two closely-related nitrogen-fixing symbiotic bacteria, Rhizobium leguminosarum biovar viciae 3841 and Rhizobium etli CFN42. These strains maintain a stable genomic core that is also common to other rhizobia species plus a very variable and significant accessory component. The chromosomes are highly syntenic, whereas plasmids are related by fewer syntenic blocks and have mosaic structures. The pairs of plasmids p42f-pRL12, p42e-pRL11 and p42b-pRL9 as well large parts of p42c with pRL10 are shown to be similar, whereas the symbiotic plasmids (p42d and pRL10 are structurally unrelated and seem to follow distinct evolutionary paths. Even though purifying selection is acting on the whole genome, the accessory component is evolving more rapidly. This component is constituted largely for proteins for transport of diverse metabolites and elements of external origin. The present analysis allows us to conclude that a heterogeneous and quickly diversifying group of plasmids co-exists in a common genomic framework.

  8. Relação entre a estrutura foliar de feijoeiro e a pré-infecção por Glomerella cingulata f.sp. phaseoli Relationship between bean leaf structure and the Glomerella cingulata f.sp. phaseoli preinfection

    Directory of Open Access Journals (Sweden)

    Vanessa de Fátima Jerba

    2005-03-01

    Full Text Available Os objetivos deste trabalho foram mensurar a área da nervura secundária e quantificar o número de tricomas da superfície foliar de cultivares de feijoeiro suscetível (Carioca, moderadamente resistente (Pérola e resistente (IAPAR-81 à antracnose, e observar as relações entre tais características com o estágio pré-infeccional. Para mensurar a área da nervura secundária, fragmentos foliares foram amostrados, fixados em FAA 50, conservados em álcool 70%, infiltrados em resina glicol-metacrilato, cortados transversalmente e corados com azul de toluidina. Os tricomas foram quantificados em impressões epidérmicas em microscópio de projeção. Na análise pré-infeccional, o material foi fixado em glutaraldeído 2,5% e processado para o estudo ao microscópio eletrônico de varredura. A cultivar resistente apresentou menor área de nervura secundária e maior pilosidade que a cultivar moderadamente resistente e a suscetível. Na cultivar resistente, por causa da menor área de nervura e maior pilosidade, o patógeno permaneceu sobre a nervura e envolvido com o tricoma. Na cultivar suscetível, com maior área de nervura e menor pilosidade, foram observadas necrose e estruturas fúngicas na superfície foliar. Há relação entre estas características foliares e a resistência à antracnose durante o estágio pré-infeccional do patógeno.The objectives of this work were to measure the secondary vein area and to quantify the trichomes of leaf surface in bean cultivar susceptible (Carioca, medium resistant (Pérola and resistant (IAPAR-81 to anthracnose, and to observe relationships between leaf blade characteristics with the preinfectional stages. In order to measure the secondary vein area, sample leaf pieces were fixed in FAA 50 and conserved in 70% alcohol. These leaf samples were infiltrated in glycol metacrylate resin, cut transversally and stained with toluidine blue. The trichomes were quantified in epidermical impressions in projection microscopy. The material for preinfection analysis was fixed in 2.5% glutaraldehyde and processed to study the scanning electron microscopy. The resistant cultivar showed minor secondary vein area and high pilosity than the cultivar medium resistant and susceptible. In the resistant cultivar, due to minor vein area and high pilosity, the pathogen stayed on the vein and involved on the trichomes. In susceptible cultivar, with high vein and minor pilosity, necrosis and fungal structure on the leaf surface were observed. The results show a relation between the minor vein and high leaf trichomes quantity with the resistance to the anthracnose in pathogen preinfection stages.

  9. Localizatlon of expansin-like protein in apoplast of pea (Pisum sativum L. root nodules during interaction with Rhizobium leguminosarum bv. viciae

    Directory of Open Access Journals (Sweden)

    Marzena Sujkowska

    2011-01-01

    Full Text Available During nodule development on pea roots, apoplast undergoes changes in activity of plant cell wall proteins such as expansins (EXPs. Because the accumulation of EXP protein has been correlated with the growth of various plant organs, we investigated using Western Blot and immunolocalization studies with antibody against PsEXP1, whether this protein was accumulated in the expanding cells of nodule. Immunoblot results indicated the presence of a 30-kDa band specific for pea root nodules. The EXP proteins content rose during growth of pea root nodules. Expansin(s protein was localized in nodule apoplast as well as in the infection thread walls. The enhanced amount of expansin-like proteins in meristematic part of nodule, root and shoot was shown. The localization of this protein in the meristematic cell walls can be related to the loosening of plant cell wall before cell enlargement. Both, plant cell enlargement and infection thread growth require activity of expansin(s. Possible involvement of EXPs in the process of pea root nodule development is also discussed.

  10. Produção e caraterização de nanocelulose bacteriana extraída de Rhizobium leguminosarum

    OpenAIRE

    Teixeira, Diana Inês Evaristo

    2017-01-01

    Dissertação de Mestrado em Engenharia Mecânica O mundo moderno deve, em grande parte, o seu desenvolvimento mecânico e tecnológico aos combustíveis fósseis. Estes tornaram-se a maior fonte de energia global, originando assim uma forte dependência. Sendo esta uma fonte não renovável e existindo uma crescente preocupação com o meio ambiente, fontes alternativas começam a ser estudadas. Os polímeros sintéticos são um subproduto dos combustíveis fósseis com uma vasta gama de aplicações, desde ...

  11. Outer membrane protein changes during bacteroid development are independent of nitrogen fixation and differ between indeterminate and determinate nodulating host plants of Rhizobium leguminosarum

    NARCIS (Netherlands)

    Roest, H.P.; Goosen-de Roo, L.; Wijffelman, C.A.; Maagd, de R.A.; Lugtenberg, B.J.J.

    1995-01-01

    The outer membrane of bacteroids contains largely decreased levels of protein antigen groups II and III in comparison with that of free-living rhizobia (R. A. de Maagd, R. de Rijk, I. H. M. Mulders, and B. J, J. Lugtenberg, J.Bacteriol, 171:1136-1142, 1989). Since we intend to study the molecular

  12. Incidence of Agrobacterium tumefaciens biovar 1 in and on ‘Paradox’ (Juglans hindsii x Juglans regia) walnut seed collected from commercial nurseries

    Science.gov (United States)

    The walnut rootstock Paradox (Juglans hindsii (Jeps) Rehder x J. regia L.) is susceptible to Agrobacterium tumefaciens (7) which often results in a high incidence of crown gall in nursery or walnut production orchards. Though A. tumefaciens is susceptible to the commonly used preplant soil fumigant...

  13. Brucellosis seroprevalence in Bali cattle with reproductive failure in South Sulawesi and Brucella abortus biovar 1 genotypes in the Eastern Indonesian archipelago

    NARCIS (Netherlands)

    Muflihanah, Hanah; Hatta, Mochammad; Rood, Ente; Scheelbeek, Pauline; Abdoel, Theresia H.; Smits, Henk L.

    2013-01-01

    Brucellosis is a major cause of infertility and reproductive failure in livestock. While cattle in the Eastern Indonesian archipelago suffers from reproductive problems information on bovine brucellosis in the region is fragmentary. The control of brucellosis requires a major and prolonged effort

  14. The pan-genome of the animal pathogen Corynebacterium pseudotuberculosis reveals differences in genome plasticity between the biovar ovis and equi strains

    DEFF Research Database (Denmark)

    Soares, Siomar C; Silva, Artur; Trost, Eva

    2013-01-01

    , Corynebacterium pseudotuberculosis infections pose a rising worldwide economic problem in ruminants. The complete genome sequences of 15 C. pseudotuberculosis strains isolated from different hosts and countries were comparatively analyzed using a pan-genomic strategy. Phylogenomic, pan-genomic, core genomic...

  15. Pheno- and genotyping of Brucella abortus biovar 5 isolated from a water buffalo (Bubalus bubalis) fetus: First case reported in the Americas.

    Science.gov (United States)

    Martínez, Diana; Thompson, Carolina; Draghi, Graciela; Canavesio, Vilma; Jacobo, Roberto; Zimmer, Patricia; Elena, Sebastián; Nicola, Ana M; de Echaide, Susana Torioni

    2014-09-17

    An isolate of Brucella spp. from an aborted water buffalo (Bubalus bubalis) fetus was characterized based on its pheno- and genotype. The phenotype was defined by carbon dioxide requirement, hydrogen sulfide production, sensitivity to thionin and basic fuchsin and agglutination with Brucella A and M monospecific antisera. The genotype was based on the amplification of the following genes: bcsp31, omp2ab, and eri and the species-specific localization of the insertion sequence IS711 in the Brucella chromosome via B. abortus-B. melitensis-B. ovis-B. suis (AMOS)-PCR. Unexpectedly, the isolate showed a phenotype different from B. abortus bv 1, the most prevalent strain in cattle in Argentina, and from vaccine strain 19, currently used in bovines and water buffaloes. Genotyping supported the phenotypic results, as the analysis of the omp2ab gene sequence showed an identical pattern to either B. abortus bv 5 or B. melitensis. Finally, the AMOS PCR generated a 1700-bp fragment from the isolate, different than those amplified from B. abortus bv 1 (498bp) and B. melitensis (731bp), confirming the presence of B. abortus bv 5. The OIE/FAO Reference Laboratory for Brucellosis confirmed this typing. This is the first report of B. abortus bv 5 from a water buffalo in the Americas. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. The cyclic AMP receptor protein, CRP, is required for both virulence and expression of the minimal CRP regulon in Yersinia pestis biovar microtus.

    Science.gov (United States)

    Zhan, Lingjun; Han, Yanping; Yang, Lei; Geng, Jing; Li, Yingli; Gao, He; Guo, Zhaobiao; Fan, Wei; Li, Gang; Zhang, Lianfeng; Qin, Chuan; Zhou, Dongsheng; Yang, Ruifu

    2008-11-01

    The cyclic AMP receptor protein (CRP) is a bacterial regulator that controls more than 100 promoters, including those involved in catabolite repression. In the present study, a null deletion of the crp gene was constructed for Yersinia pestis bv. microtus strain 201. Microarray expression analysis disclosed that at least 6% of Y. pestis genes were affected by this mutation. Further reverse transcription-PCR and electrophoretic mobility shift assay analyses disclosed a set of 37 genes or putative operons to be the direct targets of CRP, and thus they constitute the minimal CRP regulon in Y. pestis. Subsequent primer extension and DNase I footprinting assays mapped transcriptional start sites, core promoter elements, and CRP binding sites within the DNA regions upstream of pla and pst, revealing positive and direct control of these two laterally acquired plasmid genes by CRP. The crp disruption affected both in vitro and in vivo growth of the mutant and led to a >15,000-fold loss of virulence after subcutaneous infection but a pestis and, particularly, is more important for infection by subcutaneous inoculation. It can further be concluded that the reduced in vivo growth phenotype of the crp mutant should contribute, at least partially, to its attenuation of virulence by both routes of infection. Consistent with a previous study of Y. pestis bv. medievalis, lacZ reporter fusion analysis indicated that the crp deletion resulted in the almost absolute loss of pla promoter activity. The plasminogen activator encoded by pla was previously shown to specifically promote Y. pestis dissemination from peripheral infection routes (subcutaneous infection [flea bite] or inhalation). The above evidence supports the notion that in addition to the reduced in vivo growth phenotype, the defect of pla expression in the crp mutant will greatly contribute to the huge loss of virulence of this mutant strain in subcutaneous infection.

  17. Brucella melitensis Biovar 1 and Brucella abortus S19 Vaccine Strain Infections in Milkers Working at Cattle Farms in the Khartoum Area, Sudan

    NARCIS (Netherlands)

    Osman, Amira E. F.; Hassan, Abdullahi N.; Ali, Ali E.; Abdoel, Theresia H.; Smits, Henk L.

    2015-01-01

    Human brucellosis is a preventable zoonoses that may become persistent, causing, if left untreated, severe localized disease. Occupational exposure to infected animals or animal products and consumption of fresh contaminated dairy are main risk factors. One hundred farmworkers employed at two cattle

  18. Regulation of Biofilm Formation by Hfq is Influenced by Presence of Plasmid pCD1 in Yersinia Pestis Biovar Microtus

    Directory of Open Access Journals (Sweden)

    Huiying Yang

    2017-10-01

    Full Text Available Yersinia pestis synthesizes the attached biofilms in the flea gut to promotethe flea-borne transmission of this deadly pathogen. Bellows et al. reported that the posttranscriptional regulator Hfq inhibites biofilm formation in apCD1− derivative of Y. pestis CO92, however, we found that Hfq stimulates biofilm production in a microtus strain of Y. pestis with the typical plasmids, including pCD1. When we cured pCD1 from this strain, the biofilm phenotype was in accordance with that reported by Bellows et al., indicating that the unknown pCD1-associated factors modulating the regulatory pathways of Y. pestis biofilm formation. Further gene regulation experiments using relevant pCD1+ Y. pestis strains disclose that Hfq positively regulates the expression of hmsHFRS and hmsT encoding a diguanylate cyclase while negatively regulates the expression of hmsP encoding the sole phosphodiesterase. However, Hfq has no regulatory effect on the expression of hmsCDE at the mRNA and protein levels. Our results suggest that we should be cautious to make conclusion from results based on the pCD1-cured Y. pestis.

  19. Ralstonia (Pseudomonas) solanacearum race 3 (biovar 2) in surface water and natural weed hosts: First report on stinging nettle (Urtica dioica)

    NARCIS (Netherlands)

    Wenneker, M.; Verdel, M.S.W.; Groeneveld, R.M.W.; Kempenaar, C.; Beuningen, van A.R.; Janse, J.D.

    1999-01-01

    The population dynamics of the brown rot bacterium Ralstonia (Pseudomonas) solanacearum in surface water of two selected water-areas were monitored over a two-year period. In some cases during summer, high bacterial numbers (up to 106 cfu l−1) were observed. In a host plant survey a few plants of

  20. Molecular typing of Brucella species isolates from Human and livestock bloods in Isfahan province

    Directory of Open Access Journals (Sweden)

    Ebtehaj Pishva

    2015-01-01

    Conclusion: Our findings confirm abundance of B. melitensis, particularly biovar 1 in human and sheep are identical but B. abortus biovar 3 as the etiological agent of cattle brucellosis most frequently isolated in the Isfahan area.

  1. Fungitoxic properties of four crude plant extacts on fusarium ...

    African Journals Online (AJOL)

    Fungitoxic properties of four crude plant extacts on fusarium oxysporum schl. F. sp phaseoli. ... African Journal of Food, Agriculture, Nutrition and Development ... Crude plant extracts from Azadirachta indica, Tagetes minuta, Nicotiana tobacum and Vinca rosea were tested against Fusarium oxysporum Schl. F. sp. phaseoli.

  2. In-Vitro Evaluation of Fungicides and Fungicide Combinations Against Fusarium Root-Rot Fungal Pathogens of French Beans(Phaseolus Vulgaris L. c v. Monel

    International Nuclear Information System (INIS)

    Wagichunge, A.G.R; Owino, P.O; Waudo, S.W; Seif, A.A

    1999-01-01

    Laboratories studies were undertaken to evaluate In-vitro efficacy of captan, thiram, pyrazophos, triforine and metalaxyl + mancozeb fungicides against Fusarium solani (Mart.) Appel and Wollenw fsp. phaseoli (Burk) Synder and Hansen Fusarium oxysporum Schlecht fsp. phaseoli kend and Synder root-rot fungal pathogens of French beans. Five fungicides and four combinations were tested for their antifungal activity. Fungicides treatments significantly (P=0.05) inhibited mycelial growth and spore germination. Fungicides suppressed the growth of F. oxysporum fsp. Phaseoli more than that of F. solani fsp. phaseoli. All fungicides except metalaxyl + mancozeb failed to suppress sporulation of the two fungi In-vitro. In the case of thiram the sporulation capacity of F. oxysporum fsp. phaseoli 3.43 times higher than in the control. Although, no fungicides treatment was seen to inhibitor of all the three measures of fungitoxicity, the ranking of the best three fungicide treatments would be, thiram 50 + captan so > triforine > metalaxyl + mancozeb. The relatively higher inhibitory effect of fungicides on the growth of F. oxysporum Ssp. Phaseoli than that of F. solani fsp. Phaseoli suggested that F. oxysporum Esp. Phaseoli was more sensible to fungicide treatments. Such differences may reflect inherent variations in accessibility of the active toxicants within the fungal systems. The ability attributed to the low growth rate, N depletion temperature and oxygen

  3. Epiphytic bacteria from various bean genotypes and their potential ...

    African Journals Online (AJOL)

    Naturally occurring epiphytic non-pathogeni bacteria were isolated from reproductive tissue of various bean genotypes grown in the field and screened for both in vitro and in vivo antagonism to Xanthomonas axonopodis pv. phaseoli (formely X. campestris pv phaseoli). Of the 22 potential bacterial antagonists screened in ...

  4. Determination of mycoflora of pea (Pisum sativum) seeds and the ...

    African Journals Online (AJOL)

    Jane

    2011-07-06

    Jul 6, 2011 ... Mycoflora of pea seeds and the effectiveness of Rhizobium leguminosarum on important seed-borne pathogens ... seed germination and promoted the plants growth under controlled and ..... 27. Table 3. Summary of fungal inoculation test. .... and lettuce by phosphate-solubilizing Rhizobium leguminosarum.

  5. ORF Alignment: NC_002506 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002506 gi|15600953 >1cqxA 1 398 1 388 e-107 ... gb|AAF96096.1| ferrisiderophore re...ductase [Vibrio cholerae O1 biovar eltor str. ... N16961] ref|NP_232583.1| ferrisiderophore reductase... ... [Vibrio cholerae O1 biovar eltor str. N16961] ... pir||F82491 ferrisiderophore reductase

  6. Draft Genome Sequence of the Biocontrol Strain Serratia plymuthica A30, Isolated from Rotting Potato Tuber Tissue

    NARCIS (Netherlands)

    Czajkowski, R.L.; Van der Wolf, J.M.

    2012-01-01

    Serratia plymuthica A30 is a Gram-negative bacterium expressing antagonistic activity toward blackleg- and soft rot-causing Dickeya sp. biovar 3 ("Dickeya solani"). Here, we present the draft genome sequence of strain A30, which has been isolated from rotten potato tuber tissue. [KEYWORDS: biovar 3

  7. Molecular diagnosis of lymphogranuloma venereum in patients with genital ulcer disease.

    NARCIS (Netherlands)

    Sturm, P.D.J.; Moodley, P.; Govender, K.; Bohlken, L.; Vanmali, T.; Sturm, A.W.

    2005-01-01

    The detection of herpes, chancroid, and syphilis in genital ulcers is done by PCR. This is not so for lymphogranuloma venereum (LGV). We report on the use of a PCR with digestion that differentiates the LGV biovar from the trachoma biovar. Our findings suggest that the clinical description of LGV in

  8. Contributo para a melhoria de solos marginais destinados a pastagens pela aplicação de lama residual urbana, sem riscos ambientais Contribution to the improvement of degraded soils under pastures through sewage sludge application, without environmental risks

    Directory of Open Access Journals (Sweden)

    M. G. Serrão

    2009-01-01

    Full Text Available A aplicação de lamas residuais urbanas (LRU aos solos destinados a pastagens, ainda escassamente utilizada no País, contribui, com frequência, para melhorar os níveis de matéria orgânica (M.O. e de alguns nutrientes das plantas e para diminuir o risco de erosão, pelo aumento da cobertura vegetal. Todavia, a presença eventual de níveis elevados de metais pesados, compostos orgânicos poluentes e organismos patogénicos nas LRU condiciona a dose a aplicar e torna imprescindível o controlo desses factores nos solos aos quais foram incorporadas. Também o teor elevado de azoto que por vezes contêm pode inibir a actividade simbiótica do rizóbio, prejudicando a sobrevivência das leguminosas na pastagem. Neste trabalho, examinaram-se a produção de matéria seca, a composição florística e o teor de cobre (Cu na biomassa vegetal, em dois anos consecutivos de um ensaio com uma mistura pratense semeada para cortes sucessivos, instalado, no Outono de 2001, num Luvissolo Háplico de baixa fertilidade, em Mértola, ao qual foi aplicado LRU secundária proveniente da ETAR de Évora, com um elevado teor de Cu. No mesmo período, apreciou-se a evolução, na camada superficial do solo, dos teores de M.O., de alguns macronutrientes e do Cu extraível por água régia. Avaliouse, ainda, a grandeza da população rizobiana que nodula o trevo (Rhizobium leguminosarum biovar trifolii e procedeu-se à prospecção de indicadores de contaminação fecal (bactérias coliformes e enterococos. No ano seguinte à aplicação da LRU, examinou-se a evolução, no solo, de 11 compostos bifenilospoliclorados (PCBs, 13 pesticidas organoclorados e 16 hidrocarbonetos aromáticos polinucleares (PAHs. O ensaio, de blocos casualizados, teve como modalidades três níveis de LRU (L 0 = 0, L1 = 12 e L2 = 24 t/ha e duas repetições. A mistura semeada incluiu azevém anual, panasco, cinco espécies de trevo, bisserula e serradela. Além de muito maiores produ

  9. Characterization of some Brucella species from Zimbabwe by biochemical profiling and AMOS-PCR

    Directory of Open Access Journals (Sweden)

    Skjerve Eystein

    2009-12-01

    Full Text Available Abstract Background Bovine brucellosis caused by Brucella abortus is endemic in most large commercial and smallholder cattle farms of Zimbabwe, while brucellosis in other domestic animals is rare. The diagnosis of brucellosis is mainly accomplished using serological tests. However, some Brucella spp. have been isolated from clinical cases in the field and kept in culture collection but their biochemical profiles were not documented. We report biochemical profiling and AMOS-PCR characterization of some of these field isolates of Brucella originating from both commercial and smallholder cattle farming sectors of Zimbabwe. Findings Fourteen isolates of Brucella from culture collection were typed using biochemical profiles, agglutination by monospecific antisera, susceptibility to Brucella-specific bacteriophages and by AMOS-PCR that amplifies species- specific IS711. The results of the biochemical profiles for B. abortus biovar 1 (11 isolates and biovar 2 (2 isolates were consistent with those of reference strains. A single isolate from a goat originating from a smallholder mixed animal farm was identified as B. melitensis biovar 1. The AMOS-PCR produced DNA products of sizes 498 bp and 731 bp for B. abortus (biovar 1 and 2 and B. melitensis biovar 1, respectively. Conclusion We concluded that the biochemical profiles and AMOS-PCR characterization were consistent with their respective species and biovars. B. abortus biovar 1 is likely to be the predominant cause of brucellosis in both commercial and smallholder cattle farms in Zimbabwe.

  10. Recommendations of the Subcommittee on the taxonomy of Campylobacter and related bacteria

    DEFF Research Database (Denmark)

    Vandamme, P.; On, S.L.W.

    2001-01-01

    therefore discourages the use of the name C. hyoilei. (ii) The revised infrasubspecific nomenclature of Campylobacter sputorum is endorsed. C. sputorum is subdivided into C. sputorum biovar sputorum (characterized by the absence of catalase and urease activity); C. sputorum biovar faecalis (characterized...... by catalase but not urease activity); and C. sputorum biovar paraureolyticus (characterized by urease, but not catalase activity). (iii) The subcommittee points out that 'Flexispira rappini' is a taxon that is circumscribed by means of morphological criteria. It encompasses multiple Helicobacter species...

  11. Journal of Genetics | Indian Academy of Sciences

    Indian Academy of Sciences (India)

    bacterial wilt; type III effector; virulence factor; phylogenetic analysis; ... biovar3, and are the only sequenced strains from India infecting solanaceous vegetables. ... Department of Microbiology, Goa University, Taleigao Plateau 403 206, India ...

  12. Genomic Relatedness of Chlamydia Isolates Determined by Amplified Fragment Length Polymorphism Analysis

    OpenAIRE

    Meijer, Adam; Morré, Servaas A.; Van Den Brule, Adriaan J. C.; Savelkoul, Paul H. M.; Ossewaarde, Jacobus M.

    1999-01-01

    The genomic relatedness of 19 Chlamydia pneumoniae isolates (17 from respiratory origin and 2 from atherosclerotic origin), 21 Chlamydia trachomatis isolates (all serovars from the human biovar, an isolate from the mouse biovar, and a porcine isolate), 6 Chlamydia psittaci isolates (5 avian isolates and 1 feline isolate), and 1 Chlamydia pecorum isolate was studied by analyzing genomic amplified fragment length polymorphism (AFLP) fingerprints. The AFLP procedure was adapted from a previously...

  13. ORF Alignment: NC_002505 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002505 gi|15640858 >1atg0 1 226 25 253 1e-12 ... gb|AAF94004.1| accessory colonization... factor AcfC [Vibrio cholerae O1 biovar eltor ... str. N16961] ref|NP_230489.1| accessory colonization... ... factor AcfC [Vibrio cholerae O1 biovar eltor str. ... N16961] pir||F82273 accessory colonization

  14. Examination of taxonomic uncertainties surrounding Brucella abortus bv. 7 by phenotypic and molecular approaches.

    Science.gov (United States)

    Garin-Bastuji, Bruno; Mick, Virginie; Le Carrou, Gilles; Allix, Sebastien; Perrett, Lorraine L; Dawson, Claire E; Groussaud, Pauline; Stubberfield, Emma J; Koylass, Mark; Whatmore, Adrian M

    2014-03-01

    Brucella taxonomy is perpetually being reshuffled, at both the species and intraspecies levels. Biovar 7 of Brucella abortus was suspended from the Approved Lists of Bacterial Names Brucella classification in 1988, because of unpublished evidence that the reference strain 63/75 was a mixture of B. abortus biovars 3 and 5. To formally clarify the situation, all isolates previously identified as B. abortus bv. 7 in the AHVLA and ANSES strain collections were characterized by classical microbiological and multiple molecular approaches. Among the 14 investigated strains, including strain 63/75, only four strains, isolated in Kenya, Turkey, and Mongolia, were pure and showed a phenotypic profile in agreement with the former biovar 7, particularly agglutination with both anti-A/anti-M monospecific sera. These results were strengthened by molecular strategies. Indeed, genus- and species-specific methods allowed confirmation that the four pure strains belonged to the B. abortus species. The combination of most approaches excluded their affiliation with the recognized biovars (biovars 1 to 6 and 9), while some suggested that they were close to biovar 3.These assays were complemented by phylogenetic and/or epidemiological methods, such as multilocus sequence analysis (MLSA) and variable-number tandem repeat (VNTR) analysis. The results of this polyphasic investigation allow us to propose the reintroduction of biovar 7 into the Brucella classification, with at least three representative strains. Interestingly, the Kenyan strain, sharing the same biovar 7 phenotype, was genetically divergent from other three isolates. These discrepancies illustrate the complexity of Brucella taxonomy. This study suggests that worldwide collections could include strains misidentified as B. abortus bv. 7, and it highlights the need to verify their real taxonomic position.

  15. ORF Alignment: NC_005296 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_005296 gi|39936343 >1bxwA 4 170 45 261 3e-15 ... ref|YP_222291.1| Omp31-1, outer m...embrane protein, 31 kDa [Brucella abortus biovar 1 ... str. 9-941] gb|AAX74930.1| Omp31-1, outer mem...brane ... protein, 31 kDa [Brucella abortus biovar 1 str. 9-941] ... gb|AAS84568.1| Omp31b [

  16. ORF Alignment: NC_002505 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002505 gi|15641120 >1iwlA 6 181 22 197 4e-50 ... gb|AAF94266.1| outer membrane lipoproteins...|F82241 outer membrane ... lipoproteins carrier protein VC1107 [imported] ... carrier protein [Vibrio cholerae O1 ... biovar eltor str. N16961] ref|NP_230752.1| outer ... membrane lipopro...teins carrier protein [Vibrio cholerae ... O1 biovar eltor str. N16961] pir|

  17. Systemic fungicidal activity of 1,4-oxathiin derivatives.

    Science.gov (United States)

    Schmeling, B V; Kulka, M

    1966-04-29

    Treatment of pinto bean and barley seed with 1,4-oxathiin derivatives gave disease control by systemic fungicidal action of such pathogenic fungi as Uromyces phaseoli and Ustilago nuda. The two chemicals, D735 and F461, were highly specific and selective against the pathogens without injury of the hosts.

  18. Improvement of common bacterial blight resistance in South African ...

    African Journals Online (AJOL)

    Common bacterial blight (CBB) caused by Xanthomonas axonopodis pv. phaseoli is an important seed-borne disease of dry beans in South Africa. Development of resistant cultivars is considered the best control measurement for the disease. Backcross breeding was used to improve BB resistance in the small white ...

  19. Evaluating Genetic Association between Fusarium and Pythium ...

    African Journals Online (AJOL)

    Resistance to Fusarium root rot (Fusarium solani f.s.p phaseoli) has been reported in common bean (Phaseolus vulgaris L.) sources and is usually associated with Pythium root rot resistance. Pythium root rot (Pythium ultimum var ultimum) resistance is controlled by a single dominant gene, marked by a SCAR marker ...

  20. Development of a multiplex qPCR in real time for quantification and differential diagnosis of Salmonella Gallinarum and Salmonella Pullorum.

    Science.gov (United States)

    Rubio, Marcela da Silva; Penha Filho, Rafael Antonio Casarin; Almeida, Adriana Maria de; Berchieri, Angelo

    2017-12-01

    Currently there are 2659 Salmonella serovars. The host-specific biovars Salmonella Pullorum and Salmonella Gallinarum cause systemic infections in food-producing and wild birds. Fast diagnosis is crucial to control the dissemination in avian environments. The present work describes the development of a multiplex qPCR in real time using a low-cost DNA dye (SYBr Green) to identify and quantify these biovars. Primers were chosen based on genomic regions of difference (RoD) and optimized to control dimers. Primers pSGP detect both host-specific biovars but not other serovars and pSG and pSP differentiate biovars. Three amplicons showed different melting temperatures (Tm), allowing differentiation. The pSGP amplicon (97 bp) showed Tm of 78°C for both biovars. The pSG amplicon (273 bp) showed a Tm of 86.2°C for S. Gallinarum and pSP amplicon (260 bp) dissociated at 84.8°C for S. Pullorum identification. The multiplex qPCR in real time showed high sensitivity and was capable of quantifying 10 8 -10 1 CFU of these biovars.

  1. In vitro investigations on host specificity of ralstonia solanacearum among solanaceous crops and its biological control in tomato

    International Nuclear Information System (INIS)

    Hasan, S.; Haq, M. I.; Naz, F.; Tahir, M. I.

    2016-01-01

    Ralstonia solanacearum, the causal organism of bacterial wilt of solanaceous crops is a major limitation on the production of solanaceous crops worldwide. The present studies were carried out to explore the prevalence, severity, virulence and host specificity range of R. solanacearum in chili, tomato, eggplant and potato plants while biocontrol of this pathogen was studied on tomato. The isolation and the initial identification of bacterium were done on TTC and 523 media. Out of 32 isolates, 30 showed positive hypersensitive reaction (HR) with variable response and were confirmed as R. solanacearum by performing biochemical tests i.e. Gram staining, KOH, catalase oxidation, Kovacs oxidation, levan production from sucrose, lipase activity on tween 80 agar, production of fluorescent pigment, and oxidation/fermentation of glucose. Race identification studies showed that all the isolates belonged to Race 3 while biovar confirmation tests revealed that 4, 3 and 23 isolates belonged to biovar 1, biovar 2 and biovar 3, respectively. The isolates belonging to distinct biovar class were tested for their wideness of host range by challenging them to chili, tomato, eggplant and potato plants. It was observed that biovar 3 is the most aggressive and has widest host range as compared to counterparts. Biocontrol studies through antagonistic rhizobacteria resulted four antagonistic isolates; PRB10, PAP5, PAT1 and PTR6 having the highest biocontrol activity with 98.75%, 97.5%, 93.75% and 91.25% respectively. (author)

  2. MLVA genotyping of Brucella melitensis and Brucella abortus isolates from different animal species and humans and identification of Brucella suis vaccine strain S2 from cattle in China.

    Directory of Open Access Journals (Sweden)

    Hai Jiang

    Full Text Available In China, brucellosis is an endemic disease and the main sources of brucellosis in animals and humans are infected sheep, cattle and swine. Brucella melitensis (biovars 1 and 3 is the predominant species, associated with sporadic cases and outbreak in humans. Isolates of B. abortus, primarily biovars 1 and 3, and B. suis biovars 1 and 3 are also associated with sporadic human brucellosis. In this study, the genetic profiles of B. melitensis and B. abortus isolates from humans and animals were analyzed and compared by multi-locus variable-number tandem-repeat analysis (MLVA. Among the B. melitensis isolates, the majority (74/82 belonged to MLVA8 genotype 42, clustering in the 'East Mediterranean' group. Two B. melitensis biovar 1 genotype 47 isolates, belonging to the 'Americas' group, were recovered; both were from the Himalayan blue sheep (Pseudois nayaur, a wild animal. The majority of B. abortus isolates (51/70 were biovar 3, genotype 36. Ten B. suis biovar 1 field isolates, including seven outbreak isolates recovered from a cattle farm in Inner Mongolia, were genetically indistinguishable from the vaccine strain S2, based on MLVA cluster analysis. MLVA analysis provided important information for epidemiological trace-back. To the best of our knowledge, this is the first report to associate Brucella cross-infection with the vaccine strain S2 based on molecular comparison of recovered isolates to the vaccine strain. MLVA typing could be an essential assay to improve brucellosis surveillance and control programs.

  3. The rhizobium-pea symbiosis as affected by high temperatures

    NARCIS (Netherlands)

    Frings, J.F.J.

    1976-01-01

    A study has been made concerning the effect of high temperatures on the symbiosis of Rhizobium leguminosarum and pea plants (Pisum sativum). At 30°C, no nodules were found on the roots of plants growing in nutrient solution after inoculation with

  4. Effects of nano-TiO2 on the agronomically-relevant Rhizobium-legume symbiosis

    Science.gov (United States)

    The impact of nano-TiO2 on Rhizobium-legume symbiosis was studied using garden peas and the compatible bacterial partner Rhizobium leguminosarum bv. viciae 3841. Exposure to nano-TiO2 did not affect the germination of peas grown aseptically, nor did it impact the gross root structure. However, nano-...

  5. Molecular aspects of the nitrogen fixing system in pea root nodules

    International Nuclear Information System (INIS)

    Bisseling, T.

    1980-01-01

    The author considers symbiotic nitrogen fixation of Pisum sativum and Rhizobium leguminosarum. Some general aspects of nodule formation and the regulation of the proteins nitrogenase and leghemoglobin (Lb) have been studied. Synthesis of these proteins was studied by 35 SO 4 labelling of intact pea plants. The sequence of appearance of the proteins was determined with specific radioimmunoassays for each protein. (Auth.)

  6. Genetic Factors in Rhizobium Affecting the Symbiotic Carbon Costs of N2 Fixation and Host Plant Biomass Production

    DEFF Research Database (Denmark)

    Skøt, L.; Hirsch, P. R.; Witty, J. F.

    1986-01-01

    The effect of genetic factors in Rhizobium on host plant biomass production and on the carbon costs of N2 fixation in pea root nodules was studied. Nine strains of Rhizobium leguminosarum were constructed, each containing one of three symbiotic plasmids in combination with one of three different ...

  7. Phylogenetic analysis of 23S rRNA gene sequences of some ...

    African Journals Online (AJOL)

    ... glycol plus control. All isolates exhibited good drought-tolerant efficiencies at 10% PEG. While most of the isolates could not tolerate up to 20% PEG, isolates of Rlv6, Rlv9, Rlv12 and Rlv13 tolerated up to 20% PEG. Keywords: Rhizobium leguminosarum, 23S rRNA gene, phylogenetic tree, diversity and drought tolerance ...

  8. Strain identification in Rhizobium by starch gel electrophoresis of isoenzymes

    DEFF Research Database (Denmark)

    Engvild, Kjeld Christensen; Nielsen, G.

    1985-01-01

    Sonieated extracts of rhizobia, especiaUy Rhizobium leguminosarum from pea and vetch, were run in horizontal starch gel electrophoresis in the cold. The rhizobia were grown on agar on a slime suppressing substrate of tryptone-yeast extract-CaCl2 with small amounts of mannitol, sorbitol...

  9. Identification of Ononitol and O-methyl-scyllo-inositol in Pea Root Nodules

    DEFF Research Database (Denmark)

    Skøt, Leif; Egsgaard, Helge

    1984-01-01

    Ononitol (4-O-methyl-myo-inositol) and O-methyl-scyllo-inositol were identified in pea (Pisum sativum L.) root nodules formed by twoRhizobium leguminosarum strains. Ononitol was the major soluble carbohydrate in nodules formed by strain 1045 while O-methyl-scyllo-inositol and two unidentified com...

  10. Cultivar and Rhizobium Strain Effects on the Symbiotic Performance of Pea (Pisum sativum)

    DEFF Research Database (Denmark)

    Skøt, Leif

    1983-01-01

    The symbiotic performance of four pea (Pisum sativum L.) cultivars in combination with each of four strains of Rhizobium leguminosarum was studied in growth chamber experiments in order to estimate the effects of cultivars, strains and cultivar × strain interaction on the variation in dry weight, N...

  11. A novel type of DNA-binding protein interacts with a conserved sequence in an early nodulin ENOD12 promoter.

    NARCIS (Netherlands)

    Christiansen, H.; Hansen, A.C.; Vijn, I.; Pallisgaard, N.; Larsen, K.; Yang, W.C.; Bisseling, T.; Marcker, K.A.; Jensen, E.O.

    1996-01-01

    The pea genes PsENOD12A and PsENOD12B are expressed in the root hairs shortly after infection with the nitrogen-fixing bacterium Rhizobium leguminosarum bv. viciae or after application of purified Nod factors. A 199 bp promoter fragment of the PsENOD12B gene contains sufficient information for Nod

  12. Download this PDF file

    African Journals Online (AJOL)

    home

    2014-05-25

    May 25, 2014 ... Rhizobium leguminosarum bv. viciae Strains Isolated from Faba Bean. Producing ... dietary protein to the majority of population in the country. Soil acidity ...... and nitrogenase activity of pea nodules as affected by low. pH and ...

  13. Pea-root exudates and their effect upon root-nodule bacteria

    NARCIS (Netherlands)

    Egeraat, van A.W.S.M.

    1972-01-01

    The main purpose of this investigation was to study the exudation (mechanism, sites) of various compounds by roots of pea seedlings in relation to the growth of Rhizobium leguminosarum.

    Chapter 1 gives a survey of the literature pertaining to plant-root

  14. Assessment of multi-strain PGPRs biofertilization as compared to ...

    African Journals Online (AJOL)

    Sids 1, grown in clayey soil to inoculation with plant-growth promoting rhizobacteria (PGPRs) such as cyanobacteria spp; Serratia sp; Rhizobium leguminosarum bv. viciae and Bacillus polymyxa either solely or in combinations in comparison to mineral N-fertilization, Full N-dose; 75 Kg N/fed. Results revealed the superiority ...

  15. Effects of nano-ZnO on the agronomically relevant Rhizobium-legume symbiosis

    Science.gov (United States)

    The impact of nano-ZnO (nZnO) on Rhizobium-legume symbiosis was studied with garden pea and its compatible bacterial partner Rhizobium leguminosarum bv. viciae 3841. Exposure of peas to nZnO had no impact on germination, but significantly affected root length. Chronic exposure of plant to nZnO impac...

  16. Brucellosis in camels (Camelus dromedarius) in Darfur, Western Sudan.

    Science.gov (United States)

    Musa, M T; Eisa, M Z M; El Sanousi, E M; Abdel Wahab, M B; Perrett, L

    2008-01-01

    In a field outbreak of brucellosis in 21 camels mixed with cattle, sheep and goats, five camels, three of which showed clinical signs, were serologically positive. In a subsequent abattoir survey of apparently healthy camels, six animals were seropositive, albeit with titres that tended to be lower than those found in the field outbreak. Of the six seropositive slaughtered camels, five were shown to have lymph nodes (prescapular and supramammary) infected with brucellae (Brucella melitensis biovar 3, two camels; Brucella abortus biovar 6, three camels). Infection of camels with B. abortus biovar 6 had not previously been reported. Infection of the supramammary lymph nodes presents a potential hazard to those who consume raw camels' milk, a common practice in nomadic camel owners.

  17. Isolation and Molecular Characterization of Brucella Isolates in Cattle Milk in Uganda

    Directory of Open Access Journals (Sweden)

    Denis Rwabiita Mugizi

    2015-01-01

    Full Text Available Brucellosis is endemic in livestock and humans in Uganda and its transmission involves a multitude of risk factors like consumption of milk from infected cattle. To shed new light on the epidemiology of brucellosis in Uganda the present study used phenotypic and molecular approaches to delineate the Brucella species, biovars, and genotypes shed in cattle milk. Brucella abortus without a biovar designation was isolated from eleven out of 207 milk samples from cattle in Uganda. These isolates had a genomic monomorphism at 16 variable number tandem repeat (VNTR loci and showed in turn high levels of genetic variation when compared with other African strains or other B. abortus biovars from other parts of the world. This study further highlights the usefulness of MLVA as an epidemiological tool for investigation of Brucella infections.

  18. Biotyping and genotyping (MLVA16 of Brucella abortus isolated from cattle in Brazil, 1977 to 2008.

    Directory of Open Access Journals (Sweden)

    Sílvia Minharro

    Full Text Available Brucellosis is a worldwide distributed zoonosis that causes important economic losses to animal production. In Brazil, information on the distribution of biovars and genotypes of Brucella spp. is scarce or unavailable. This study aimed (i to biotype and genotype 137 Brazilian cattle isolates (from 1977 to 2008 of B. abortus and (ii to analyze their distribution. B. abortus biovars 1, 2 and 3 (subgroup 3b were confirmed and biovars 4 and 6 were first described in Brazil. Genotyping by the panel 1 revealed two groups, one clustering around genotype 40 and another around genotype 28. Panels 2A and 2B disclosed a high diversity among Brazilian B. abortus strains. Eighty-nine genotypes were found by MLVA16. MLVA16 panel 1 and 2 showed geographic clustering of some genotypes. Biotyping and MLVA16 genotyping of Brazilian B. abortus isolates were useful to better understand the epidemiology of bovine brucellosis in the region.

  19. Chlamydia trachomatis Mip-like protein

    DEFF Research Database (Denmark)

    Lundemose, AG; Rousch, DA; Birkelund, Svend

    1992-01-01

    A 27 kDa Chlamydia trachomatis Mip-like protein with homology of a 175-amino-acid C-terminal fragment to the surface-exposed Legionella pneumophila mip-gene product has previously been described. In this paper the entire chlamydia Mip-like sequence of C. trachomatis serovar L2 (lymphogranuloma...... venereum (LGV) biovar) is presented. The sequence shows high similarity to the legionella Mip protein and its C-terminal region, like that of the legionella Mip, has high amino acid similarity to eukaryotic and prokaryotic FK506-binding proteins. The chlamydial mip-like gene was detected by polymerase...... chain reaction (PCR) in other C. trachomatis serovars and by sequencing of the mip-like genes of serovars B and E (trachoma biovar) was shown to be highly conserved within the two major biovars of C. trachomatis. Monoclonal and polyclonal antibodies raised against the recombinant Mip-like protein failed...

  20. Differential phenotyping of Brucella species using a newly developed semi-automated metabolic system

    Directory of Open Access Journals (Sweden)

    Appel Bernd

    2010-10-01

    Full Text Available Abstract Background A commercial biotyping system (Taxa Profile™, Merlin Diagnostika testing the metabolization of various substrates by bacteria was used to determine if a set of phenotypic features will allow the identification of members of the genus Brucella and their differentiation into species and biovars. Results A total of 191 different amines, amides, amino acids, other organic acids and heterocyclic and aromatic substrates (Taxa Profile™ A, 191 different mono-, di-, tri- and polysaccharides and sugar derivates (Taxa Profile™ C and 95 amino peptidase- and protease-reactions, 76 glycosidase-, phosphatase- and other esterase-reactions, and 17 classic reactions (Taxa Profile™ E were tested with the 23 reference strains representing the currently known species and biovars of Brucella and a collection of 60 field isolates. Based on specific and stable reactions a 96-well "Brucella identification and typing" plate (Micronaut™ was designed and re-tested in 113 Brucella isolates and a couple of closely related bacteria. Brucella species and biovars revealed characteristic metabolic profiles and each strain showed an individual pattern. Due to their typical metabolic profiles a differentiation of Brucella isolates to the species level could be achieved. The separation of B. canis from B. suis bv 3, however, failed. At the biovar level, B. abortus bv 4, 5, 7 and B. suis bv 1-5 could be discriminated with a specificity of 100%. B. melitensis isolates clustered in a very homogenous group and could not be resolved according to their assigned biovars. Conclusions The comprehensive testing of metabolic activity allows cluster analysis within the genus Brucella. The biotyping system developed for the identification of Brucella and differentiation of its species and biovars may replace or at least complement time-consuming tube testing especially in case of atypical strains. An easy to handle identification software facilitates the

  1. Direct quantification of fungal DNA from soil substrate using real-time PCR.

    Science.gov (United States)

    Filion, Martin; St-Arnaud, Marc; Jabaji-Hare, Suha H

    2003-04-01

    Detection and quantification of genomic DNA from two ecologically different fungi, the plant pathogen Fusarium solani f. sp. phaseoli and the arbuscular mycorrhizal fungus Glomus intraradices, was achieved from soil substrate. Specific primers targeting a 362-bp fragment from the SSU rRNA gene region of G. intraradices and a 562-bp fragment from the F. solani f. sp. phaseoli translation elongation factor 1 alpha gene were used in real-time polymerase chain reaction (PCR) assays conjugated with the fluorescent SYBR(R) Green I dye. Standard curves showed a linear relation (r(2)=0.999) between log values of fungal genomic DNA of each species and real-time PCR threshold cycles and were quantitative over 4-5 orders of magnitude. Real-time PCR assays were applied to in vitro-produced fungal structures and sterile and non-sterile soil substrate seeded with known propagule numbers of either fungi. Detection and genomic DNA quantification was obtained from the different treatments, while no amplicon was detected from non-seeded non-sterile soil samples, confirming the absence of cross-reactivity with the soil microflora DNA. A significant correlation (Pgenomic DNA of F. solani f. sp. phaseoli or G. intraradices detected and the number of fungal propagules present in seeded soil substrate. The DNA extraction protocol and real-time PCR quantification assay can be performed in less than 2 h and is adaptable to detect and quantify genomic DNA from other soilborne fungi.

  2. ORF Alignment: NC_002505 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_002505 gi|15640861 >1bxwA 8 146 21 199 4e-04 ... gb|AAF94007.1| accessory colonization... factor AcfA [Vibrio cholerae O1 biovar eltor ... str. N16961] ref|NP_230492.1| accessory colonization... ... factor AcfA [Vibrio cholerae O1 biovar eltor str. ... N16961] gb|AAK20801.1| accessory colonization... factor ... AcfA [Vibrio cholerae] gb|AAK20771.1| accessory ... colonization... factor AcfA [Vibrio cholerae] pir||JC4087 ... accessory colonization factor

  3. ORF Alignment: NC_003318 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_003318 gi|17989189 >1bxwA 4 170 40 240 2e-16 ... gb|AAN33618.1| outer membrane protein Omp3...1 [Brucella suis 1330] gb|AAS84567.1| ... Omp31 [Brucella cetaceae] gb|AAS84566.1| Omp31 [Br...ucella ... cetaceae] gb|AAS84565.1| Omp31 [Brucella pinnipediae] ... gb|AAS84564.1| Omp31 [Bru...nsis] ... gb|AAL27297.1| outer membrane protein Omp31 [Brucella ... melitensis biovar Neotomae...] gb|AAL27290.1| outer ... membrane protein Omp31 [Brucella melitensis biovar Suis] ... gb|AAL

  4. THERMOTOLERANT Campylobacter SPECIES ISOLATED FROM PSITTACIFORMES IN THE PERUVIAN AMAZON REGION

    Directory of Open Access Journals (Sweden)

    Alvaro TRESIERRA-AYALA

    1998-07-01

    Full Text Available Foi determinada a freqüência de isolamento de campylobacters termotolerantes em Psittaciformes silvestres capturados na região amazônica do Peru. Campylobacters foram isolados em 10/142 (7.0% dos animais estudados, sendo C. jejuni subsp. jejuni biovar I (6/10 o mais freqüente, seguido de C. coli biovar II (2/10, C. lari não foi isolado. Os resultados sugerem que estas aves podem ser importantes reservatórios destas bactérias.

  5. "Comparación de algunas propiedades de las lectinas de semilla y de raíz de haba

    Directory of Open Access Journals (Sweden)

    Yolanda De Navarro

    2010-05-01

    Full Text Available Se aislaron las lectinas de semilla y de las paredes celulares de la raíz de haba. La lectina de raíz se purificó parcialmente por Sephadex G-25 obteniéndose resultados que sugieren la presencia de dos lectinas. El comportamiento en filtración por gel, diálisis, electroforesis inhibición por carbohidratos de la eritroaglutinación, acción aglutinante frente a la cepa B de R. leguminosarum cultivada en medio Bishop. capacidad aglutinante de cepas homologas y heterólogas de las dos lectinas demostraron que ellas son diferentes. La lectina de raíz parece ser más específica, ya que solo aglutinó a R. leguminosarum y R. spp. (caupi, mientras que la Favina presentó actividad bactoaglutinante con cepas nodulantes como con no nodulantes de diferentes especies de Rhizobium.

  6. Degradation of the Herbicide Glyphosate by Members of the Family Rhizobiaceae

    OpenAIRE

    Liu, C.-M.; McLean, P. A.; Sookdeo, C. C.; Cannon, F. C.

    1991-01-01

    Several strains of the family Rhizobiaceae were tested for their ability to degrade the phosphonate herbicide glyphosate (isopropylamine salt of N-phosphonomethylglycine). All organisms tested (seven Rhizobium meliloti strains, Rhizobium leguminosarum, Rhizobium galega, Rhizobium trifolii, Agrobacterium rhizogenes, and Agrobacterium tumefaciens) were able to grow on glyphosate as the sole source of phosphorus in the presence of the aromatic amino acids, although growth on glyphosate was not a...

  7. AcEST: DK956784 [AcEST

    Lifescience Database Archive (English)

    Full Text Available tolB OS=Rhizobium leguminosarum bv.... 38 0.042 sp|A8F168|TOLB_RICM5 Protein tolB OS=Rickettsia massiliae (s...PDGARVSFES 309 >sp|A8F168|TOLB_RICM5 Protein tolB OS=Rickettsia massiliae (strain Mtu5) GN=tolB PE=3 SV=1 Le

  8. Symbiotic Activity of Pea (Pisum sativum) after Application of Nod Factors under Field Conditions

    OpenAIRE

    Siczek, Anna; Lipiec, Jerzy; Wielbo, Jerzy; Kidaj, Dominika; Szarlip, Paweł

    2014-01-01

    Growth and symbiotic activity of legumes are mediated by Nod factors (LCO, lipo-chitooligosaccharides). To assess the effects of application of Nod factors on symbiotic activity and yield of pea, a two-year field experiment was conducted on a Haplic Luvisol developed from loess. Nod factors were isolated from Rhizobium leguminosarum bv. viciae strain GR09. Pea seeds were treated with the Nod factors (10−11 M) or water (control) before planting. Symbiotic activity was evaluated by measurement...

  9. Characteristics and phylogeny of Bacillus cereus strains isolated from Maari, a traditional West African food condiment

    DEFF Research Database (Denmark)

    Thorsen, Line; Kando, Christine Kere; Sawadogo, Hagrétou

    2015-01-01

    light on the succession and pathogenic potential of B. cereus species in traditional West African food condiment and clarifies their phylogenetic relatedness to B. cereus biovar anthracis. Future implementation of GMP and HACCP and development of starter cultures for controlled Maari fermentations...

  10. Population dynamics of Streptococcus mitis in its natural habitat

    DEFF Research Database (Denmark)

    Hohwy, J.; Reinholdt, Jesper; Kilian, Mogens

    2001-01-01

    showed that the ability to persist was not distributed at random in the S. mitis biovar 1 population. However, neither immunoglobulin A1 protease activity nor the ability to bind alpha-amylase from saliva was a preferential characteristic of persistent genotypes. In contrast to current concepts of climax...

  11. Efficacy of antibiotic treatment and test-based culling strategies for eradicating brucellosis in commercial swine herds

    NARCIS (Netherlands)

    Dieste-Pérez, L.; Frankena, K.; Blasco, J.M.; Muñoz, P.M.; Jong, de M.C.M.

    2016-01-01

    Swine brucellosis caused by Brucella suis biovar 2 is an emerging disease in continental Europe. Without effective vaccines being available, the European Food Safety Authority (EFSA) recommends the full depopulation of infected herds as the only strategy to eradicate B. suis outbreaks. Using data

  12. Efficacy of antibiotic treatment and test-based culling strategies for eradicating brucellosis in commercial swine herds

    NARCIS (Netherlands)

    Dieste-Pérez, L.; Frankena, K.; Blasco, J. M.; Muñoz, P. M.; De Jong, M. C M

    2016-01-01

    Swine brucellosis caused by Brucella suis biovar 2 is an emerging disease in continental Europe. Without effective vaccines being available, the European Food Safety Authority (EFSA) recommends the full depopulation of infected herds as the only strategy to eradicate B. suis outbreaks. Using data

  13. Differentiation between serological responses to Brucella suis and Yersinia enterocolitica serotype O : 9 after natural or experimental infection in pigs

    DEFF Research Database (Denmark)

    Jungersen, Gregers; Sørensen, Vibeke; Giese, Steen Bjørck

    2006-01-01

    with responses of B. suis biovar 2-inoculated pigs. FPSR were limited to 2-9 weeks post-YeO:9 inoculation, while B. suis-infected pigs were test-positive throughout the 21-week period of investigation. Although YeO:9-inoculated pigs exhibited FPSR in Brucella tests for a limited period of time, the serological...

  14. "HOOF-Print" Genotyping and Haplotype Inference Discriminates among Brucella spp Isolates From a Small Spatial Scale

    Science.gov (United States)

    We demonstrate that the “HOOF-Print” assay provides high power to discriminate among Brucella isolates collected on a small spatial scale (within Portugal). Additionally, we illustrate how haplotype identification using non-random association among markers allows resolution of B. melitensis biovars ...

  15. Draft Genome Sequence of the Biocontrol Strain Serratia plymuthica A30, Isolated from Rotting Potato Tuber Tissue

    NARCIS (Netherlands)

    Czajkowski, R.L.; Wolf, van der J.M.

    2012-01-01

    Serratia plymuthica A30 is a Gram-negative bacterium expressing antagonistic activity toward blackleg- and soft rot-causing Dickeya sp. biovar 3 (“Dickeya solani”). Here, we present the draft genome sequence of strain A30, which has been isolated from rotten potato tuber tissue

  16. ORF Alignment: NC_004310 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_004310 gi|23502301 >1p3dA 28 461 27 463 e-121 ... ref|YP_222116.1| MurC, UDP-N-ace...tylmuramate--alanine ligase [Brucella abortus biovar ... 1 str. 9-941] gb|AAX74755.1| MurC, ...

  17. ORF Alignment: NC_003317 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_003317 gi|17986863 >1p3dA 28 461 27 463 e-121 ... ref|YP_222116.1| MurC, UDP-N-ace...tylmuramate--alanine ligase [Brucella abortus biovar ... 1 str. 9-941] gb|AAX74755.1| MurC, ...

  18. Chlamydia trachomatis Mip-like protein

    DEFF Research Database (Denmark)

    Lundemose, AG; Rousch, DA; Birkelund, Svend

    1992-01-01

    venereum (LGV) biovar) is presented. The sequence shows high similarity to the legionella Mip protein and its C-terminal region, like that of the legionella Mip, has high amino acid similarity to eukaryotic and prokaryotic FK506-binding proteins. The chlamydial mip-like gene was detected by polymerase...

  19. ORF Alignment: NC_005810 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_005810 gi|45441793 >1u07A 1 89 164 251 4e-26 ... ref|NP_669352.1| energy transduce...] ... ref|NP_993332.1| TonB protein [Yersinia pestis biovar ... Medievalis str. 91001] gb|AAM85603.1| energy

  20. ORF Alignment: NC_004088 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_004088 gi|22125929 >1u07A 1 89 164 251 4e-26 ... ref|NP_669352.1| energy transduce...] ... ref|NP_993332.1| TonB protein [Yersinia pestis biovar ... Medievalis str. 91001] gb|AAM85603.1| energy

  1. ORF Alignment: NC_003143 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_003143 gi|16122423 >1u07A 1 89 164 251 4e-26 ... ref|NP_669352.1| energy transduce...] ... ref|NP_993332.1| TonB protein [Yersinia pestis biovar ... Medievalis str. 91001] gb|AAM85603.1| energy

  2. Investigation of the bacteriophage community in induced lysates of undefined mesophilic mixed-strain DL-cultures using classical and metagenomic approaches

    DEFF Research Database (Denmark)

    Muhammed, Musemma K.; Olsen, Mette L.; Kot, Witold

    2018-01-01

    To investigate the notion that starter cultures can be a reservoir of bacteriophages (phages) in the dairy environment, strains of three DL-starters (undefined mesophilic mixed-strain starters containing Lactococcus lactis subsp. lactis biovar. diacetylactis and Leuconostoc species) were selected...

  3. Potential for biocontrol of hairy root disease by a Paenibacillus clade

    NARCIS (Netherlands)

    Bosmans, Lien; de Bruijn, Irene; Gerards, S.; Moerkens, Rob; Van Looveren, Lore; Wittemans, Lieve; Van Calenberge, Bart; Paeleman, Anneleen; Van Kerckhove, Stef; Rozenski, Jef; de Mot, Rene; Rediers, Hans; Raaijmakers, J.M.; Lievens, Bart

    2017-01-01

    Rhizogenic Agrobacterium biovar 1 is the causative agent of hairy root disease (HRD) in the hydroponic cultivation of tomato and cucumber causing significant losses in marketable yield. In order to prevent and control the disease chemical disinfectants such as hydrogen peroxide or hypochlorite are

  4. Improving cost-effectiveness of brown rot control: the value of bio-economic modelling

    NARCIS (Netherlands)

    Breukers, A.; Werf, van der W.; Mourits, M.C.M.; Oude Lansink, A.G.J.M.

    2007-01-01

    Since 1995, the Dutch potato production chain has been hit by several outbreaks of brown rot, a quarantine disease caused by Ralstonia solanacearum race 3, biovar 2. To avoid establishment of brown rot in the potato production chain and avert the consequences on potato export, the Dutch government

  5. ORF Alignment: NC_006155 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_006155 gi|51596443 >1u07A 1 89 164 251 4e-26 ... ref|NP_669352.1| energy transduce...] ... ref|NP_993332.1| TonB protein [Yersinia pestis biovar ... Medievalis str. 91001] gb|AAM85603.1| energy

  6. Multi-locus variable-number tandem repeat analysis of Chinese Brucella strains isolated from 1953 to 2013.

    Science.gov (United States)

    Tian, Guo-Zhong; Cui, Bu-Yun; Piao, Dong-Ri; Zhao, Hong-Yan; Li, Lan-Yu; Liu, Xi; Xiao, Pei; Zhao, Zhong-Zhi; Xu, Li-Qing; Jiang, Hai; Li, Zhen-Jun

    2017-05-02

    Brucellosis was a common human and livestock disease caused by Brucella strains, the category B priority pathogens by the US Center for Disease Control (CDC). Identified as a priority disease in human and livestock populations, the increasing incidence in recent years in China needs urgent control measures for this disease but the molecular background important for monitoring the epidemiology of Brucella strains at the national level is still lacking. A total of 600 Brucella isolates collected during 60 years (from 1953 to 2013) in China were genotyped by multiple locus variable-number tandem repeat analysis (MLVA) and the variation degree of MLVA11 loci was calculated by the Hunter Gaston Diversity Index (HGDI) values. The charts and map were processed by Excel 2013, and cluster analysis and epidemiological distribution was performed using BioNumerics (version 5.1). The 600 representative Brucella isolates fell into 104 genotypes with 58 singleton genotypes by the MLVA11 assay, including B. melitensis biovars 2 and 3 (five main genotypes), B. abortus biovars 1 and 3 (two main genotypes), B. suis biovars 1 and 3 (three main genotypes), and B. canis (two main genotypes) respectively. While most B. suis biovar 1 and biovar 3 were respectively found in northern provinces and southern provinces, B. melitensis and B. abortus strains were dominant in China. Canine Brucellosis was only found in animals without any human cases reported. Eight Brucellosis epidemic peaks emerged during the 60 years between 1953 and 2013: 1955 - 1959, 1962 - 1969, 1971 - 1975, 1977 - 1983, 1985 - 1989, 1992 - 1997, 2000 - 2008 and 2010 - 2013 in China. Brucellosis has its unique molecular epidemiological patterns with specific spatial and temporal distribution according to MLVA. IDOP-D-16-00101.

  7. Genotyping and phylogenetic analysis of Yersinia pestis by MLVA: insights into the worldwide expansion of Central Asia plague foci.

    Directory of Open Access Journals (Sweden)

    Yanjun Li

    Full Text Available BACKGROUND: The species Yersinia pestis is commonly divided into three classical biovars, Antiqua, Medievalis, and Orientalis, belonging to subspecies pestis pathogenic for human and the (atypical non-human pathogenic biovar Microtus (alias Pestoides including several non-pestis subspecies. Recent progress in molecular typing methods enables large-scale investigations in the population structure of this species. It is now possible to test hypotheses about its evolution which were proposed decades ago. For instance the three classical biovars of different geographical distributions were suggested to originate from Central Asia. Most investigations so far have focused on the typical pestis subspecies representatives found outside of China, whereas the understanding of the emergence of this human pathogen requires the investigation of strains belonging to subspecies pestis from China and to the Microtus biovar. METHODOLOGY/PRINCIPAL FINDINGS: Multi-locus VNTR analysis (MLVA with 25 loci was performed on a collection of Y. pestis isolates originating from the majority of the known foci worldwide and including typical rhamnose-negative subspecies pestis as well as rhamnose-positive subspecies pestis and biovar Microtus. More than 500 isolates from China, the Former Soviet Union (FSU, Mongolia and a number of other foci around the world were characterized and resolved into 350 different genotypes. The data revealed very close relationships existing between some isolates from widely separated foci as well as very high diversity which can conversely be observed between nearby foci. CONCLUSIONS/SIGNIFICANCE: The results obtained are in full agreement with the view that the Y. pestis subsp. pestis pathogenic for humans emerged in the Central Asia region between China, Kazakhstan, Russia and Mongolia, only three clones of which spread out of Central Asia. The relationships among the strains in China, Central Asia and the rest of the world based on the MLVA

  8. Human Brucellosis in Maghreb: Existence of a Lineage Related to Socio-Historical Connections with Europe

    Science.gov (United States)

    Lounes, Nedjma; Cherfa, Moulay-Ali; Le Carrou, Gilles; Bouyoucef, Abdellah; Jay, Maryne; Garin-Bastuji, Bruno; Mick, Virginie

    2014-01-01

    Despite control/eradication programs, brucellosis, major worldwide zoonosis due to the Brucella genus, is endemic in Northern Africa and remains a major public health problem in the Maghreb region (Algeria/Morocco/Tunisia). Brucella melitensis biovar 3 is mostly involved in human infections and infects mainly small ruminants. Human and animal brucellosis occurrence in the Maghreb seems still underestimated and its epidemiological situation remains hazy. This study summarizes official data, regarding Brucella melitensis infections in Algeria, from 1989 to 2012, with the purpose to provide appropriate insights concerning the epidemiological situation of human and small ruminant brucellosis in Maghreb. Algeria and Europe are closely linked for historical and economical reasons. These historical connections raise the question of their possible impact on the genetic variability of Brucella strains circulating in the Maghreb. Other purpose of this study was to assess the genetic diversity among Maghreb B. melitensis biovar 3 strains, and to investigate their possible epidemiological relationship with European strains, especially with French strains. A total of 90 B. melitensis biovar 3 Maghreb strains isolated over a 25 year-period (1989–2014), mainly from humans, were analysed by MLVA-16. The obtained results were compared with genotypes of European B. melitensis biovar 3 strains. Molecular assays showed that Algerian strains were mainly distributed into two distinct clusters, one Algerian cluster related to European sub-cluster. These results led to suggest the existence of a lineage resulting from socio-historical connections between Algeria and Europe that might have evolved distinctly from the Maghreb autochthonous group. This study provides insights regarding the epidemiological situation of human brucellosis in the Maghreb and is the first molecular investigation regarding B. melitensis biovar 3 strains circulating in the Maghreb. PMID:25517901

  9. Investigation of radiosensitivity and growth dynamics for callus tissues Crepis Capillaris, Haplopappus gracilis, Phasolium vulgaris exposed to gamma radiation

    International Nuclear Information System (INIS)

    Gatsek, Eh.; Glinkova, E.; Ismailova, Eh.N.

    1983-01-01

    Radiosensitivity of three kinds of callus tissues (Crepis capillaris, Haplopappus gracilis, Phasolium vulgaris) manifested in the change of fresh weight after γ-irradiation has been investigated. Irradiated callus arowth showed decrease with increasing doses. It is shown that the radiosensitivity of ''young'' callus tissues is determined by the kind of the plant. Callus of Phaseolis has been found to have the highest radioresistance, while that of Crepis has the lowest one. Radiosensitivity of ''old'' callus tissues is the same for all kinds. Potential mechanism of radiosensitivity of callus tissUes are discussed

  10. Fungicide selective for basidiomycetes.

    Science.gov (United States)

    Edgington, L V; Walton, G S; Miller, P M

    1966-07-15

    Concentrations of 2,3-dihydro-5-carboxanilido-6-methyl-1,4-oxathiin lower than 8 parts per million prevented mycelial growth of a number of Basidiomycetes. By contrast, mycelial growth of various other fungi-Phycomycetes, Ascomycetes, and Deuteromycetes-was 50 percent inhibited only by concentrations of 32 ppm or higher. Two exceptions to this pattern of selective fungitoxicity were found:an isolate of Rhizoctonia solani was not as sensitive as other Basidiomycetes, and the deuteromycete Verticillium alboatrum was inhibited by lower concentrations than affected other fungi in this group. Spore germination of two Basidiomycetes, Uromyces phaseoli and Ustilago nuda, was inhibited 95 percent or more at 10 ppm.

  11. Type of prey influences biology and consumption rate of Orius insidiosus (Say (Hemiptera, Anthocoridae

    Directory of Open Access Journals (Sweden)

    Simone M. Mendes

    2002-01-01

    Full Text Available Generalist predators are capable of consuming different types of prey, and as each prey may have distinct nutritional values, each may have a distinct impact on the biology of the predator. Therefore, the aim of this study was to determine how the consumption of different prey influences certain biological characteristics and the predatory capacity of Orius insidiosus (Say. The investigation was performed in climatic chamber at 25 ±1 ºC, RH 70 ± 10% and fotophase 12. Eggs of Anagasta kuehniella (Zeller, adults of Caliothrips phaseoli (Hood and nymphs of Aphis gossypii Glover were used as prey and were provided daily ad libitum for all the mobile stages of the predator. The results showed that biological parameters of O. insidiosus are affected differently depending on the type of prey ingested. The development time of the nymphal stage was 13.1, 11.23 and 10.25 days for O. insidiosus feeding on eggs of A. kuehniella, nymphs of A. gossypii and adults of C. phaseoli, respectively. Longevity was five times larger for adults fed on eggs of A. kuehniella (56.25 days compared to that of adults that preyed on nymphs of A. gossypii (11.44 days, and four times larger when the prey were adults of C. phaseoli (13.58 days. The consumption of eggs of A. kuehniella by predator females resulted in a shorter pre-oviposition period (3.2 days and a longer oviposition period (44.4 days when compared to the consumption of other types of prey. In addition, fecundity was increased with the consumption of eggs of A. kuehniella (195.25 eggs laid / female when compared to feeding on the other prey, C. phaseoli (70.00 eggs laid / female or A. gossypii (22.50 eggs laid / female. However, the consumption of aphids was larger (148.28 nymphs/ nymphal stage than that of thrips (74.10 thrips / nymphal stage or eggs of A. kuehniella (37.03 eggs /nymphal stage for all of the nymphal stages of the predator. The results indicate that the eggs of A. kuehniella are the type of

  12. EFEITO ANTIBIÓTICO DO PRÓPOLIS SOBRE BACTÉRIAS FITOPATOGÊNICAS

    OpenAIRE

    BIANCHINI, L.; BEDENDO, I.P.

    1998-01-01

    O efeito antibiótico de extrato aquoso de própolis, em várias concentrações, foi avaliado para cinco espécies de bactérias fitopatogênicas. Agrobacterium tumefaciens, Clavibacter michiganensis subsp. michiganensis e Xanthomonas axonopodis pv. phaseoli foram completamente inibidas em meio de cultura contendo 10% de extrato de própolis. Erwinia chrysanthemi foi parcialmente inibida, enquanto Pseudomonas syringae pv. tabaci se mostrou insensível ao extrato, desenvolvendo colônias idênticas àquel...

  13. Brucella 'HOOF-Prints': strain typing by multi-locus analysis of variable number tandem repeats (VNTRs

    Directory of Open Access Journals (Sweden)

    Halling Shirley M

    2003-07-01

    Full Text Available Abstract Background Currently, there are very few tools available for subtyping Brucella isolates for epidemiological trace-back. Subtyping is difficult because of the genetic homogeneity within the genus. Sequencing of the genomes from three Brucella species has facilitated the search for DNA sequence variability. Recently, hypervariability among short tandem repeat sequences has been exploited for strain-typing of several bacterial pathogens. Results An eight-base pair tandem repeat sequence was discovered in nine genomic loci of the B. abortus genome. Eight loci were hypervariable among the three Brucella species. A PCR-based method was developed to identify the number of repeat units (alleles at each locus, generating strain-specific fingerprints. None of the loci exhibited species- or biovar-specific alleles. Sometimes, a species or biovar contained a specific allele at one or more loci, but the allele also occurred in other species or biovars. The technique successfully differentiated the type strains for all Brucella species and biovars, among unrelated B. abortus biovar 1 field isolates in cattle, and among B. abortus strains isolated from bison and elk. Isolates from the same herd or from short-term in vitro passage exhibited little or no variability in fingerprint pattern. Sometimes, isolates from an animal would have multiple alleles at a locus, possibly from mixed infections in enzootic areas, residual disease from incomplete depopulation of an infected herd or molecular evolution within the strain. Therefore, a mixed population or a pool of colonies from each animal and/or tissue was tested. Conclusion This paper describes a new method for fingerprinting Brucella isolates based on multi-locus characterization of a variable number, eight-base pair, tandem repeat. We have named this technique "HOOF-Prints" for Hypervariable Octameric Oligonucleotide Finger-Prints. The technique is highly discriminatory among Brucella species, among

  14. Detecção de Ralstonia solanacearum em tubérculos de batata através de PCR qualitativa e quantitativa

    OpenAIRE

    Adriana de Andrade Figueiró

    2008-01-01

    Ralstonia solanacearum é uma bactéria transmitida por tubérculos-semente. As biovares 1 (raça 1) e 2 (raça 3) estão associadas à batata (Solanum tuberosum) e apresentam características epidemiológicas diferentes. A certificação de tubérculos-semente constitui-se num processo essencial para o manejo da murcha bacteriana, mas depende de métodos específicos e sensíveis de detecção deste patógeno. Este trabalho objetivou (i) projetar oligonucleotídeos iniciadores para diferenciação das biovares d...

  15. Physiological, Biochemical and Genetic Characteristics of Ralstonia solanacearum Strains Isolated from Pepper Plants in Korea

    Directory of Open Access Journals (Sweden)

    Young Kee Lee

    2013-12-01

    Full Text Available Totally sixty three bacteria were isolated from lower stems showing symptoms of bacterial wilt on pepper plants in 14 counties of 7 provinces, Korea. The isolates showed strong pathogenicity on red pepper (cv. Daewang and tomato (cv. Seogwang seedlings. All virulent bacteria were identified as Ralstonia solanacearum based on colony types, physiological and biochemical tests and polymerase chain reaction (PCR. All R. solanacearum isolates from peppers were race 1. The bacterial isolates consisted of biovar 3 (27% and biovar 4 (73%. Based on polymorphic PCR bands generated by repetitive sequence (rep-PCR, the 63 R. solanacearum isolates were divided into 12 groups at 70% similarity level. These results will be used as basic materials for resistant breeding program and efficient control against bacterial wilt disease of pepper.

  16. GenBank blastx search result: AK104368 [KOME

    Lifescience Database Archive (English)

    Full Text Available AK104368 001-035-E04 AF361941.1 Brucella melitensis biovar Abortus urease accessory protein UreD (ureD), ure...ase gamma subunit UreA (ureA), urease beta subunit UreB (ureB), urease alpha subunit UreC (ureC), urease... accessory protein UreE (ureE), urease accessory protein UreF (ureF), and urease access

  17. Identifikasi Brucella abortus Isolat Lokal dengan Brucella abortus Strain Specific-Polymerase Chain Reaction (IDENTIFICATION OF LOCAL ISOLATES OF BRUCELLA ABORTUS USING BRUCELLA ABORTUS STRAIN SPECIFIC-POLYMERASE CHAIN REACTION ASSAY)

    OpenAIRE

    Susan Maphilindawati Noor; Pratiwi Pujilestari Sudarmono; Asmarani Kusumawati; Anis Karuniawati

    2014-01-01

    Brucella abortus Strain Specific-Polymerase Chain Reaction (BaSS-PCR) is a single multiplex PCRtechnique which able to identify and differentiate between Brucella abortus field strains (biovar 1, 2, and4), B. abortus vaccine strains, Brucella species, and non-Brucella species. In this study, BaSS-PCR wasapplied to identify local isolates of B. abortus in order to investigate the B. abortus strains that infectedcattle in Indonesia. Fifty local strains of B.abortus isolated from infected cattle...

  18. Complete Genome Sequence of Yersinis pestis Strains Antiqua and Nepa1516: Evidence of Gene Reduction in an Emerging Pathogen

    Energy Technology Data Exchange (ETDEWEB)

    Chain, Patrick S [ORNL; Hu, Ping [Lawrence Berkeley National Laboratory (LBNL); Malfatti, Stephanie [Lawrence Livermore National Laboratory (LLNL); Radnedge, Lyndsay [Lawrence Livermore National Laboratory (LLNL); Larimer, Frank W [ORNL; Vergez, Lisa [Lawrence Livermore National Laboratory (LLNL); Worsham, Patricia [U.S. Army Medical Research Institute of Infectious Diseases; Chu, May C [Centers for Disease Control and Prevention; Anderson, Gary L [Lawrence Berkeley National Laboratory (LBNL)

    2006-01-01

    Yersinia pestis, the causative agent of bubonic and pneumonic plagues, has undergone detailed study at the molecular level. To further investigate the genomic diversity among this group and to help characterize lineages of the plague organism that have no sequenced members, we present here the genomes of two isolates of the ''classical'' antiqua biovar, strains Antiqua and Nepal516. The genomes of Antiqua and Nepal516 are 4.7 Mb and 4.5 Mb and encode 4,138 and 3,956 open reading frames, respectively. Though both strains belong to one of the three classical biovars, they represent separate lineages defined by recent phylogenetic studies. We compare all five currently sequenced Y. pestis genomes and the corresponding features in Yersinia pseudotuberculosis. There are strain-specific rearrangements, insertions, deletions, single nucleotide polymorphisms, and a unique distribution of insertion sequences. We found 453 single nucleotide polymorphisms in protein-coding regions, which were used to assess the evolutionary relationships of these Y. pestis strains. Gene reduction analysis revealed that the gene deletion processes are under selective pressure, and many of the inactivations are probably related to the organism's interaction with its host environment. The results presented here clearly demonstrate the differences between the two biovar antiqua lineages and support the notion that grouping Y. pestis strains based strictly on the classical definition of biovars (predicated upon two biochemical assays) does not accurately reflect the phylogenetic relationships within this species. A comparison of four virulent Y. pestis strains with the human-avirulent strain 91001 provides further insight into the genetic basis of virulence to humans.

  19. Differences in two-component signal transduction proteins among the genus Brucella: implications for host preference and pathogenesis

    DEFF Research Database (Denmark)

    Binnewies, Tim Terence; Ussery, David; Lavín, JL

    2010-01-01

    . anthropi lacks orthologs of the Brucella TCSs NodVW, TceSR and TcfSR, suggesting that these TCS proteins could be necessary for the adaptation of Brucella as an intracellular pathogen. This genomic analysis revealed the presence of a differential distribution of TCS pseudogenes among Brucella species....... Moreover, there were also differences in TCS pseudogenes between strains belonging to the same Brucella species, and in particular between B. suis biovars 1 and 2....

  20. Lactic Acid Bacteria from Kefir Increase Cytotoxicity of Natural Killer Cells to Tumor Cells

    OpenAIRE

    Takuya Yamane; Tatsuji Sakamoto; Takenori Nakagaki; Yoshihisa Nakano

    2018-01-01

    The Japanese fermented beverage, homemade kefir, contains six lactic acid bacteria: Lactococcus. lactis subsp. Lactis, Lactococcus. lactis subsp. Cremoris, Lactococcus. Lactis subsp. Lactis biovar diacetylactis, Lactobacillus plantarum, Leuconostoc meseuteroides subsp. Cremoris and Lactobacillus casei. In this study, we found that a mixture of the six lactic acid bacteria from kefir increased the cytotoxicity of human natural killer KHYG-1 cells to human chronic myelogenous leukemia K562 cell...

  1. Infections following the application of leeches: two case reports and review of the literature

    Directory of Open Access Journals (Sweden)

    Maetz Benjamin

    2012-10-01

    Full Text Available Abstract Introduction Since the 1980s, leeches have been ingeniously used in the management of venous flap congestion. The presence of anticoagulative substances in their saliva improves the blood drainage. Their digestive tract contains several bacterial species, the main ones being Aeromonas hydrophila and Aeromonas veronii biovar sobria, which contribute to the digestion of ingested blood. These bacteria can be the cause of infections. Case presentation We report two cases of septicemia related to Aeromonas veronii biovar sobria that presented after leeches had been applied to congested transverse rectus abdominis myocutaneous flaps for delayed mammary reconstructions. Patient number 1 was a 55-year-old Caucasian woman who underwent a delayed breast reconstruction procedure. On the sixth postoperative day she showed a clinical presentation of septicemia. Aeromonas veronii biovar sobria was identified in the patient’s skin and blood bacteriological samples. Her fever ceased after 4 days of antibiotic treatment. Patient number 2 was a 56-year-old Caucasian woman who underwent a delayed breast reconstruction procedure. On the seventh postoperative day we noticed that she showed a clinical presentation of septicemia. Aeromonas veronii biovar sobria was identified in the patient’s blood cultures and local bacteriological samples. An antibiogram showed resistance to amoxicillin/clavulanic acid. Her fever ceased on the eleventh postoperative day after 4 days of antibiotic treatment. Conclusion The rate of infection after application of leeches is not negligible. The concentration of Aeromonas inside the digestive tracts of leeches largely decreases when the patient is under antibiotic therapy. These germs are sensitive to third-generation cephalosporins and fluoroquinolones and resistant to amoxicillin/clavulanic acid. We recommend preventive treatment based on classical measures of asepsis and on oral antibioprophylaxy with a fluoroquinolone

  2. Detection of virulence-associated genes in Brucella melitensis ...

    African Journals Online (AJOL)

    The current study involved detection of three virulence genes (bvfA, virB, ure) by PCR in 52 isolates of Brucella melitensis biovar 3, recovered from different animal species (28 sheep, 10 goats, 9 cattle and 5 buffaloes). Of the 52 B. melitensis strains; 48 (92.3%) isolates carried bvfA genes, 51 (98.1%) isolates had virB genes ...

  3. Inhibition of Lipid A Biosynthesis as the Primary Mechanism of CHIR-090 Antibiotic Activity in Escherichia coli

    Science.gov (United States)

    Barb, Adam W.; McClerren, Amanda L.; Snehelatha, Karnem; Reynolds, C. Michael; Zhou, Pei; Raetz, Christian R.H.

    2009-01-01

    The deacetylation of UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine (UDP-3-O-acyl-GlcNAc) by LpxC is the committed reaction of lipid A biosynthesis. CHIR-090, a novel N-aroyl-l-threonine hydroxamic acid, is a potent, slow, tight-binding inhibitor of the LpxC deacetylase from the hyperthermophile Aquifex aeolicus, and it has excellent antibiotic activity against P. aeruginosa and E. coli, as judged by disk diffusion assays. We now report that CHIR-090 is also a two-step slow, tight-binding inhibitor of Escherichia coli LpxC with Ki = 4.0 nM, Ki* = 0.5 nM, k5 = 1.9 min-1 and k6 = 0.18 min-1. CHIR-090 at low nM levels inhibits LpxC orthologues from diverse Gram-negative pathogens, including Pseudomonas aeruginosa, Neisseria meningitidis, and Helicobacter pylori. In contrast, CHIR-090 is a relatively weak competitive and conventional inhibitor (lacking slow, tight-binding kinetics) of LpxC from Rhizobium leguminosarum (Ki = 340 nM), a Gram-negative plant endosymbiont that is resistant to this compound. The KM (4.8 μM) and the kcat (1.7 s-1) of R. leguminosarum LpxC with UDP-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine as the substrate are similar to values reported for E. coli LpxC. R. leguminosarum LpxC therefore provides a useful control for validating LpxC as the primary target of CHIR-090 in vivo. An E. coli construct in which the chromosomal lpxC gene is replaced by R. leguminosarum lpxC is resistant to CHIR-090 up to 100 μg/mL, or 400 times above the minimal inhibitory concentration for wild-type E. coli. Given its relatively broad spectrum and potency against diverse Gram-negative pathogens, CHIR-090 is an excellent lead for the further development of new antibiotics targeting the lipid A pathway. PMID:17335290

  4. Insecticides and soil microorganisms

    International Nuclear Information System (INIS)

    Mostafa, I.Y.; Helal, G.A.; El-Zawahry, Y.A.

    1991-01-01

    The enzymatic degradation of C-labelled methomyl in the culture media of Rihizobium leguminosarum (isolated from Vicia feba nodules), Aspergillus fumigatus, A.niger, Fusarium oxysporum and Penicillium notatum (isolated from Allium cepa rhizophere) was studied. The results revealed that 36.5, 28.6,27.9,32.5 and 20.1% of the applied doses were metabolized into hydrolytic products by the above organisms, respectively. The thin layer chromatographic analysis of the chloroform and aqueous fractions of the media showed that C-labelled methomyl underwent a process of breakdown by these organisms into some metabolites.4 tab

  5. Sexual recombination in Colletotrichum lindemuthianum occurs on a fine scale.

    Science.gov (United States)

    Souza, E A; Camargo, O A; Pinto, J M A

    2010-09-08

    Glomerella cingulata f. sp phaseoli is the sexual phase of the fungus Colletotrichum lindemuthianum, the causal agent of common bean anthracnose. This fungus is of great concern, because it causes large economic losses in common bean crops. RAPD markers of five populations of G. cingulata f. sp phaseoli from two Brazilian states were analyzed to determine if this population possesses the sexual reproductive potential to generate the genetic variation that is observed in this phytopathogen. We identified 128 polymorphic bands, amplified by 28 random primers. The estimates of genetic similarity in this analysis ranged from 0.43 to 1.00, and the dendrogram generated from analysis of all genotypes displayed five principal groups, coinciding with the five populations. Genetic differentiation was observed between the populations (GST=0.6455); 69% of the overall observed genetic variation was between individual populations and 31% of the variance was within the sub-populations. We identified significant levels of linkage disequilibrium in all populations. However, the values of the disequilibrium ranged from low to moderate, indicating that this pathogen maintains a genetic structure consistent with sexual reproduction. The mean contribution of sexual reproduction was determined by comparison of the amplitudes of genetic similarity of isolates from sexual and asexual phases. These results support the hypothesis that recombination plays an important role in determining the amplitude of variability in this pathogen population and that this determination occurs on a fine scale.

  6. Toxicity of twenty-two plant essential oils against pathogenic bacteria of vegetables and mushrooms.

    Science.gov (United States)

    Todorović, Biljana; Potočnik, Ivana; Rekanović, Emil; Stepanović, Miloš; Kostić, Miroslav; Ristić, Mihajlo; Milijašević-Marčić, Svetlana

    2016-12-01

    ASBTRACT Toxicity of twenty-two essential oils to three bacterial pathogens in different horticultural systems: Xanthomonas campestris pv. phaseoli (causing blight of bean), Clavibacter michiganensis subsp. michiganensis (bacterial wilt and canker of tomato), and Pseudomonas tolaasii (causal agent of bacterial brown blotch on cultivated mushrooms) was tested. Control of bacterial diseases is very difficult due to antibiotic resistance and ineffectiveness of chemical products, to that essential oils offer a promising alternative. Minimal inhibitory and bactericidal concentrations are determined by applying a single drop of oil onto the inner side of each plate cover in macrodilution assays. Among all tested substances, the strongest and broadest activity was shown by the oils of wintergreen (Gaultheria procumbens), oregano (Origanum vulgare), and lemongrass (Cymbopogon flexuosus. Carvacrol (64.0-75.8%) was the dominant component of oregano oils, while geranial (40.7%) and neral (26.7%) were the major constituents of lemongrass oil. Xanthomonas campestris pv. phaseoli was the most sensitive to plant essential oils, being susceptible to 19 oils, while 11 oils were bactericidal to the pathogen. Sixteen oils inhibited the growth of Clavibacter michiganensis subsp. michiganensis and seven oils showed bactericidal effects to the pathogen. The least sensitive species was Pseudomonas tolaasii as five oils inhibited bacterial growth and two oils were bactericidal. Wintergreen, oregano, and lemongrass oils should be formulated as potential biochemical bactericides against different horticultural pathogens.

  7. Mitochondrial genome sequences reveal evolutionary relationships of the Phytophthora 1c clade species.

    Science.gov (United States)

    Lassiter, Erica S; Russ, Carsten; Nusbaum, Chad; Zeng, Qiandong; Saville, Amanda C; Olarte, Rodrigo A; Carbone, Ignazio; Hu, Chia-Hui; Seguin-Orlando, Andaine; Samaniego, Jose A; Thorne, Jeffrey L; Ristaino, Jean B

    2015-11-01

    Phytophthora infestans is one of the most destructive plant pathogens of potato and tomato globally. The pathogen is closely related to four other Phytophthora species in the 1c clade including P. phaseoli, P. ipomoeae, P. mirabilis and P. andina that are important pathogens of other wild and domesticated hosts. P. andina is an interspecific hybrid between P. infestans and an unknown Phytophthora species. We have sequenced mitochondrial genomes of the sister species of P. infestans and examined the evolutionary relationships within the clade. Phylogenetic analysis indicates that the P. phaseoli mitochondrial lineage is basal within the clade. P. mirabilis and P. ipomoeae are sister lineages and share a common ancestor with the Ic mitochondrial lineage of P. andina. These lineages in turn are sister to the P. infestans and P. andina Ia mitochondrial lineages. The P. andina Ic lineage diverged much earlier than the P. andina Ia mitochondrial lineage and P. infestans. The presence of two mitochondrial lineages in P. andina supports the hybrid nature of this species. The ancestral state of the P. andina Ic lineage in the tree and its occurrence only in the Andean regions of Ecuador, Colombia and Peru suggests that the origin of this species hybrid in nature may occur there.

  8. Genetic diversity of Ralstonia solanacearum strains from China assessed by PCR-based fingerprints to unravel host plant- and site-dependent distribution patterns.

    Science.gov (United States)

    Xue, Qing-Yun; Yin, Yan-Ni; Yang, Wei; Heuer, Holger; Prior, Philippe; Guo, Jian-Hua; Smalla, Kornelia

    2011-03-01

    Bacterial wilt caused by Ralstonia solanacearum is a serious threat to crop production in China. A collection of 319 R. solanacearum strains isolated from 14 different diseased host plants collected in 15 Chinese provinces was investigated by BOX fingerprints in order to test the influence of the site and the host plant on their genetic diversity. Phylotype, fliC-RFLP patterns and biovar were determined for all strains and the sequevar for 39 representative strains. The majority of strains belonged to the Asian phylotype I, shared identical fliC-RFLP patterns and were assigned to four biovars (bv3:123; bv4:162; bv5:3; and bv6:11). Twenty strains were phylotype II, assigned to biovar 2, and had distinct fliC-RFLP patterns. BOX-PCR fingerprints generated from the genomic DNA of each strain revealed a high diversity of the phylotype I strains, where 28 types of BOX fingerprints could be distinguished. While many BOX clusters comprised isolates from different provinces and several host plants, some groups contained isolates that were plant or site specific. All phylotype II isolates originating from 10 provinces belonged to sequevar 1 and displayed identical BOX patterns as the potato brown rot strains from various regions of the world. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  9. Phylogeny and Classification of Yersinia pestis Through the Lens of Strains From the Plague Foci of Commonwealth of Independent States

    Directory of Open Access Journals (Sweden)

    Vladimir V. Kutyrev

    2018-05-01

    Full Text Available The established phylogeny of the etiological agent of plague, Yersinia pestis, is not perfect, as it does not take into account the strains from numerous natural foci of Commonwealth of Independent States (CIS. We have carried out PCR and SNP typing of 359 strains and whole genome sequencing of 51 strains from these plague foci and determined the phylogenetic diversity of the strains circulating here. They belong to 0.ANT3, 0.ANT5, 2.ANT3, 4.ANT branches of antique biovar, 2.MED0, 2.MED1 branches of medieval biovar and to 0.PE2, 0.PE4a. 0.PE4h, 0.PE4t branches. Based on the studies of 178 strains from 23 plague foci of CIS countries, it was determined that the population structure of 2.MED strains is subdivided into Caucasian–Caspian and Central Asian–Chinese branches. In Central-Caucasian high-mountain plague foci in the Russian Federation (RF the most deeply diverged branch of medieval biovar, 2.MED0, has been found. With the data obtained, the current population structure of Y. pestis species has been refined. New subspecies classification is developed, comprising seven subspecies: pestis, caucasica (0.PE2, angolica (0.PE3, central asiatica (0.PE4, tibetica (0.PE7, ulegeica (0.PE5, and qinghaica (0.PE10.

  10. Assessment of Genetic Diversity of Zoonotic Brucella spp. Recovered from Livestock in Egypt Using Multiple Locus VNTR Analysis

    Directory of Open Access Journals (Sweden)

    Ahmed M. S. Menshawy

    2014-01-01

    Full Text Available Brucellosis is endemic in most parts of Egypt, where it is caused mainly by Brucella melitensis biovar 3, and affects cattle and small ruminants in spite of ongoing efforts devoted to its control. Knowledge of the predominant Brucella species/strains circulating in a region is a prerequisite of a brucellosis control strategy. For this reason a study aiming at the evaluation of the phenotypic and genetic heterogeneity of a panel of 17 Brucella spp. isolates recovered from domestic ruminants (cattle, buffalo, sheep, and goat from four governorates during a period of five years (2002–2007 was carried out using microbiological tests and molecular biology techniques (PCR, MLVA-15, and sequencing. Thirteen strains were identified as B. melitensis biovar 3 while all phenotypic and genetic techniques classified the remaining isolates as B. abortus (n=2 and B. suis biovar 1 (n=2. MLVA-15 yielded a high discriminatory power (h=0.801, indicating a high genetic diversity among the B. melitensis strains circulating among domestic ruminants in Egypt. This is the first report of the isolation of B. suis from cattle in Egypt which, coupled with the finding of B. abortus, suggests a potential role of livestock as reservoirs of several zoonotic Brucella species in the region.

  11. Isolation and identification of bovine Brucella isolates from Pakistan by biochemical tests and PCR.

    Science.gov (United States)

    Ali, Shahzad; Ali, Qurban; Melzer, Falk; Khan, Iahtasham; Akhter, Shamim; Neubauer, Heinrich; Jamal, Syed M

    2014-01-01

    Brucellosis is endemic in bovines in Pakistan. The Brucella species and biovars involved, however, are unknown. The objectives of the present study were to isolate and characterize brucellae from seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes which had recently aborted. The seropositive milk samples, aborted fetuses, and vaginal swabs of cattle and buffaloes were collected from the Potohar Plateau, Pakistan. Isolation of brucellae was done on modified Farrell's serum dextrose agar. Isolates were characterized by conventional biotyping methods, while molecular typing was done by genus (B4/B5) and species-specific (Brucella abortus, Brucella melitensis, Brucella ovis, and Brucella suis) polymerase chain reaction (PCR). A total of 30 isolates were recovered from milk (n = 5), aborted fetuses (n = 13), and vaginal swabs (n = 12). Most isolates were from cattle (56.7 %). All of them were identified as B. abortus biovar 1 based on conventional biotyping methods and genus and species-specific PCR. This preliminary study provides the first report on the prevalence of B. abortus biovar 1 in cattle and buffaloes in Pakistan.

  12. Complete Genome Sequence of Yersinia pestis Strains Antiqua andNepal516: Evidence of Gene Reduction in an Emerging Pathogen

    Energy Technology Data Exchange (ETDEWEB)

    Chain, Patrick S.G.; Hu, Ping; Malfatti, Stephanie A.; Radnedge,Lyndsay; Larimer, Frank; Vergez, Lisa M.; Worsham, Patricia; Chu, May C.; Andersen, Gary L.

    2006-01-16

    Yersinia pestis, the causative agent of bubonic andpneumonicplague, has undergone detailed study at the molecular level. Tofurther investigate the genomic diversity among this group and to helpcharacterize lineages of the plague organism that have no sequencedmembers, we present here the genomes of two isolates of the "classical"Antiqua biovar, strains Antiqua and Nepal516. The genomes of Antiqua andNepal516 are 4.7 Mb and 4.5 Mb and encode 4,138 and 3,956 open readingframes respectively. Though both strains belong to one of the threeclassical biovars, they represent separate lineages defined by recentphylogenetic studies. We compare all five currently sequenced Y. pestisgenomes and the corresponding features in Y. pseudotuberculosis. Thereare strain-specific rearrangements, insertions, deletions, singlenucleotide polymorphisms and a unique distribution of insertionsequences. We found 453 single nucleotide polymorphisms in protein codingregions, which were used to assess evolutionary relationships of these Y.pestis strains. Gene reduction analysis revealed that the gene deletionprocesses are under selective pressure and many of the inactivations areprobably related to the organism s interaction with its host environment.The results presented here clearly demonstrate the differences betweenthe two Antiqua lineages and support the notion that grouping Y. pestisstrains based strictly on the classical definition of biovars (predicatedupon two biochemical assays) does not accurately reflect the phylogeneticrelationships within this species. Comparison of four virulent Y. pestisstrains with the human-avirulent strain 91001 provides further insightinto the genetic basis of virulence to humans.

  13. Anal lymphogranuloma venereum infection screening with IgA anti-Chlamydia trachomatis-specific major outer membrane protein serology.

    Science.gov (United States)

    de Vries, Henry J C; Smelov, Vitaly; Ouburg, Sander; Pleijster, Jolein; Geskus, Ronald B; Speksnijder, Arjen G C L; Fennema, Johannes S A; Morré, Servaas A

    2010-12-01

    Anal lymphogranuloma venereum (LGV) infections, caused by Chlamydia trachomatis biovar L (Ct+/LGV+), are endemic among men who have sex with men (MSM). Anal non-LGV biovar Ct infections (Ct+/LGV-) can be eradicated with 1 week doxycycline, whereas Ct+/LGV+ infections require 3-week doxycycline. To differentiate Ct+/LGV+ from Ct+/LGV- infections, biovar-specific Nucleic Acid Amplification Test (NAAT) are standard, but also expensive and laborious. A chlamydia-specific serological assay could serve as an alternative test. MSM were screened for anal Ct+/LGV+ and Ct+/LGV- infections with a commercial nonspecific NAAT and an in house biovar L-specific NAAT. Serum samples were evaluated with chlamydia-specific anti-Major Outer Membrane Protein (MOMP) and antilipopolysaccharide assays of IgA and IgG classes. Asymptomatic patients were identified as: (1) no anal complaints or (2) no microscopic inflammation (i.e., <10 leucocytes per high power field in anal smears). The best differentiating assay was subsequently evaluated in 100 Ct+/LGV+ and 100 Ct+/LGV- MSM using different cut-off points. The anti-MOMP IgA assay was the most accurate to differentiate Ct+/LGV+ (n = 42) from Ct+/LGV- (n = 19) with 85.7% sensitivity (95% confidence interval [CI], 72.2-93.3) and 84.2% specificity (95% CI, 62.4-94.5), even among asymptomatic patients. In a population comprising 98 Ct+/LGV+ and 105 Ct+/LGV- patients, the anti-MOMP IgA assay scored most accurate when the cut-off point was set to 2.0 with 75.5% (95% CI, 65.8-83.6) sensitivity and 74.3% (95% CI, 64.8-82.3) specificity. The IgA anti-MOMP assay can identify a considerable proportion of the (asymptomatic) anal LGV infections correctly. Yet, biovar L-specific NAAT are still the preferred diagnostic tests in clinical settings.

  14. A ribosomal RNA gene intergenic spacer based PCR and DGGE fingerprinting method for the analysis of specific rhizobial communities in soil.

    Science.gov (United States)

    de Oliveira, Valéria Maia; Manfio, Gilson Paulo; da Costa Coutinho, Heitor Luiz; Keijzer-Wolters, Anneke Christina; van Elsas, Jan Dirk

    2006-03-01

    A direct molecular method for assessing the diversity of specific populations of rhizobia in soil, based on nested PCR amplification of 16S-23S ribosomal RNA gene (rDNA) intergenic spacer (IGS) sequences, was developed. Initial generic amplification of bacterial rDNA IGS sequences from soil DNA was followed by specific amplification of (1) sequences affiliated with Rhizobium leguminosarum "sensu lato" and (2) R. tropici. Using analysis of the amplified sequences in clone libraries obtained on the basis of soil DNA, this two-sided method was shown to be very specific for rhizobial subpopulations in soil. It was then further validated as a direct fingerprinting tool of the target rhizobia based on denaturing gradient gel electrophoresis (DGGE). The PCR-DGGE approach was applied to soils from fields in Brazil cultivated with common bean (Phaseolus vulgaris) under conventional or no-tillage practices. The community fingerprints obtained allowed the direct analysis of the respective rhizobial community structures in soil samples from the two contrasting agricultural practices. Data obtained with both primer sets revealed clustering of the community structures of the target rhizobial types along treatment. Moreover, the DGGE profiles obtained with the R. tropici primer set indicated that the abundance and diversity of these organisms were favoured under NT practices. These results suggest that the R. leguminosarum-as well as R. tropici-targeted IGS-based nested PCR and DGGE are useful tools for monitoring the effect of agricultural practices on these and related rhizobial subpopulations in soils.

  15. Rhizobium Strain Effects on Yield and Bleeding Sap Amino Compounds in Pisum sativum

    DEFF Research Database (Denmark)

    Rosendahl, Lis

    1984-01-01

    Bleeding sap composition, dry matter production and N distribution in pea (P. sativum L. cv. Bodil) grown with and without nitrate and nodulated with either R. leguminosarum strain 128c53 or strain 1044 were compared. Nitrate increased the total dry matter production of both symbioses, but decrea......Bleeding sap composition, dry matter production and N distribution in pea (P. sativum L. cv. Bodil) grown with and without nitrate and nodulated with either R. leguminosarum strain 128c53 or strain 1044 were compared. Nitrate increased the total dry matter production of both symbioses...... relative to the total N-accumulation was greater with strain 128c53 due to a higher production of nodule tissue. The root bleeding sap of the symbiosis with the greater yield (strain 1044) contained high levels of asparagine and aspartic acid. In the 128c53 symbiosis, glutamine plus homoserine accounted...... for a higher percentage of the organic solutes transporting newly assimilated N from the root system than in the association with 1044. The Rhizobium strain effect on amino compound composition of the bleeding sap may indicate an influence of the bacteroids on either the N-assimilatory enzyme system...

  16. On the influence of abiotic stress conditions on growth of barley and bean and their predisposition for pathogens

    Energy Technology Data Exchange (ETDEWEB)

    Oerke, E.C.; Schoenbeck, F.

    1986-01-01

    Shorttime changes of environmental conditions stressed barley and bean and affected plant growth and their predisposition for various pathogens. Moderate stress intensities as low or high temperatures, water or light deficits, increased the susceptibility to Erysiphe graminis var. hordei or Uromyces phaseoli and reduced disease level of spot blotch caused by Cochliobolus sativus, respectively. There was only little effect on plant growth in that case. Intensive stress as a result of combinations of unfavorable environmental conditions or longtime continuance of moderate stress reduced the plant growth and turned the predisposing effect to the opposite: after the treatment, plants were more resistent to diseases caused by biotrophic fungi, whereas there was increased susceptibility to the perthotrophic fungus. High intensities of fertilization acted as an additional stress and intensified the plant reaction to environmental alterations. The variation of the predisposition is discussed in relation to stress intensity.

  17. Linhagens de feijoeiro comum com fenótipos agronômicos favoráveis e resistência ao crestamento bacteriano comum e antracnose Common bean lines with suitable agronomic phenotypes and resistant to bacterial blight and anthracnose

    Directory of Open Access Journals (Sweden)

    Joaquim Geraldo Cáprio da Costa

    2003-10-01

    Full Text Available O crestamento bacteriano comum e a antracnoseincitados por Xanthomonas axonopodis pv. phaseoli (Smith Vauterin, Hoste, Kesters & Swings e Colletotrichum lindemuthianum (Sacc. & Magn. Scrib., respectivamente, podem reduzir consideravelmente a produtividade do feijoeiro comum (Phaseolus vulgaris L.. Conduziu-se este trabalho com o objetivo de obter linhagens com resistência conjunta ao crestamento bacteriano comum e à antracnose, com características agronômicas desejáveis. Foram obtidas 35 populações F2 provenientes de cruzamentos simples entre linhagens com resistência ao crestamento bacteriano comum proveniente de P. acutifolius A. Gray e linhagens resistentes à antracnose e com características agronômicas favoráveis. As populações segregantes de F2 a F5 foram inoculadas com o patótipo 89 de C. lindemuthianum com uma concentração de 1,2 x 10(6 conídios/mL e submetidas à seleção massal modificada. As plantas que apresentaram grau superior a 4 foram eliminadas. Na geração F5, foi realizada seleção individual das plantas remanescentes para obter as famílias F6. As famílias F6 foram avaliadas mediante inoculação com os patótipos 55, 89, 95 e 453 de C. lindemuthianum. As famílias F7 resistentes à antracnose foram inoculadas em casa-de-vegetação por incisão das folhas primárias com tesoura mergulhada em uma suspensão de 5 x 10(7 ufc do isolado XpCNF 15 de X. axonopodis pv. phaseoli. As linhagens resistentes foram avaliadas em quatro experimentos de campo. Foram selecionadas 17 linhagens com alto rendimento de grãos, resistentes ao crestamento bacteriano comum e à antracnose, com boa altura de inserção das vagens inferiores e resistentes ao acamamento.Common bacterial blight and anthracnose diseases, incited by Xanthomonas axonopodis pv. phaseoli (Smith Vauterin, Hoste, Kesters & Swings and Colletotrichum lindemuthianum (Sacc. & Magn. Scrib., respectively, are major constrains for dry bean (Phaseolus vulgaris L

  18. Micro-autoradiographic studies on host-parasite interactions. Pt. 1

    International Nuclear Information System (INIS)

    Mendgen, K.; Heitefuss, R.

    1975-01-01

    Tritium labeled uredospores of Uromyces phaseoli were produced be feeding the host, Phaseolus vulgaris, with 2 H-orotic acid. These spores were allowed to germinate on and to penetrate into a bean leaf. 24 hrs after inoculation, the bean rust had formed the first haustorium. All fungal structures, including the fungus walls, were heavily labeled. No label could be detected in the cells that had come into contact with the hyphae. In the infected host cell, the haustorium was labeled heavily, but the sheath around the haustorium and the host cell remained free of label. These results indicate that no detectable amounts of label leach from the bean rust into the host at this stage of infection although it is known that the rust takes up many metabolites. Since the sheath remains free of label and all fungal structures are evenly labeled, it is concluded that the sheath is formed by the host. (orig.) [de

  19. ANTIFUNGAL AND CYTOTOXIC ACTIVITIES OF FIVE TRADITIONALLY USED INDIAN MEDICINAL PLANTS

    Directory of Open Access Journals (Sweden)

    Adhikarimayum Haripyaree

    2013-02-01

    Full Text Available Hexane, Methanol and Distilled water extracts of five Indian Medicinal plants viz., Mimosa pudica L, Vitex trifolia Linn, Leucas aspera Spreng, Centella asiatica (L Urban and Plantago major Linn belonging to different families were subjected to preliminary antimicrobial screening against six standard organisms viz., Ceratocystis paradoxa, Aspergillus niger, Penicillium citrinum, Macrophomina phaseoli, Trichoderma viride and Rhizopus nigricans. To evaluate antifungal activity agar well diffusion method was used. In addition LD50 of the same plant extracts were determined by using Range test on Mus musculus for cytotoxic activity. Methanolic extract of M. pudica showed the highest and significant inhibitory effect against some fungal species. Again, methanolic extract of M. pudica displayed the greatest cytotoxic activity.

  20. Induced mutants in beans and peas resistant to rust

    International Nuclear Information System (INIS)

    Fadl, F.A.M.

    1983-01-01

    Beans (Phaseolus vulgaris) and peas (Pisum sativum) are important leguminous vegetable crops in Egypt. The area planted with beans is about 40,000 acres and peas 22,000 acres. These crops suffer from several diseases, particularly rusts, (Uromyces phaseoli/Uromyces pisi), which are mainly spread in northern Egypt. In our mutation induction programme we used 60 Co gamma rays and ethyl methane sulphonate (EMS). Bean and pea seeds were soaked in water for two hours before exposure to 8, 10 and 12 krad. For chemical treatments, bean and pea seeds were soaked in water for eight hours and then treated with 0.5 and 1.5% EMS for four hours. The M 1 was cultivated in 1978

  1. Determinação da variabilidade em isolados de Colletotrichum lindemuthianum por meio de marcadores morfológicos e culturais Determination of variability in isolates of Colletotrichum lindemuthianum based on morphological and cultural markers

    Directory of Open Access Journals (Sweden)

    Breno Oliveira de Souza

    2007-08-01

    Full Text Available Colletotrichum lindemuthianum (teleomorfo Glomerella cingulata f. sp. phaseoli apresenta ampla variabilidade genética, demonstrada por suas características morfológicas. Com este trabalho, objetivou-se caracterizar, por meio de marcadores morfológicos, diferentes isolados de C. lindemuthianum e identificar marcadores morfológicos com uso potencial em análises genéticas. Foram avaliados os seguintes caracteres morfológicos e culturais: cor e textura das colônias, compatibilidade vegetativa e sexual, índice de velocidade de crescimento micelial (IVCM, diâmetro colonial (DC, capacidade de esporulação (CE, dimensões e formas conidiais, dimensões dos ascósporos, formação de estruturas reprodutivas e formação de anastomoses entre hifas e conídios. Os resultados obtidos demonstraram que os isolados de C. lindemuthianum possuem ampla variabilidade genética para todas as características avaliadas e que a forma do conídio pode ser usada como marcador morfológico em análises genéticas.Colletotrichum lindemuthianum (teleomorfo Glomerella cingulata f. sp. phaseoli presents wide genetic variability, demonstrated by its morphological traits. The objective of this study was to characterize morphological markers in different isolates of C. lindemuthianum and, to identify useful morphological markers in genetic analyses. The following morphological and cultural traits were evaluated: color and texture of the colonies, vegetative and sexual compatibility, micelial growth index (MGI, colonial diameter (CD, esporulation capacity (EC, conidia dimensions and form, ascospores dimensions and formation of reproductive structures. The data showed wide genetic variability for all traits and that conidial form can be used as morphological marker in genetic analysis.

  2. EFEITO ANTIBIÓTICO DO PRÓPOLIS SOBRE BACTÉRIAS FITOPATOGÊNICAS

    Directory of Open Access Journals (Sweden)

    L. BIANCHINI

    1998-01-01

    Full Text Available O efeito antibiótico de extrato aquoso de própolis, em várias concentrações, foi avaliado para cinco espécies de bactérias fitopatogênicas. Agrobacterium tumefaciens, Clavibacter michiganensis subsp. michiganensis e Xanthomonas axonopodis pv. phaseoli foram completamente inibidas em meio de cultura contendo 10% de extrato de própolis. Erwinia chrysanthemi foi parcialmente inibida, enquanto Pseudomonas syringae pv. tabaci se mostrou insensível ao extrato, desenvolvendo colônias idênticas àquelas observadas em meios livres de própolis. Concentrações menores não foram suficientes para exercer um efeito antimicrobiano desejável sobre as bactérias pertencentes a todas as espécies testadas. Resultados idênticos foram obtidos quando própolis foi incorporado ao meio antes ou após a autoclavagem, demonstrando que a substância ativa presente no extrato não é termosensível. Foi demonstrado, portanto, o uso potencial do própolis como antibiótico para o controle de bactérias fitopatogênicas.The antibiotic effect of aqueous extract of propolis was evaluated in vitro against five species of bacteria. The growth of Agrobacterium tumefaciens, Clavibacter michiganensis subsp. michiganensis and Xanthomonas axonopodis pv. phaseoli was completely inhibited in a medium containing 10% of propolis. Erwinia chrysanthemi was partially inhibited and Pseudomonas syringae pv. tabaci was not affected. Lower concentration of the aqueous extract was not effective to promote a desirable antibiotic action against all bacterial species tested. The active ingredient present in the propolis was thermostable. The potential use of the propolis to control phytopathogenic bacteria was demonstrated.

  3. Reliable identification at the species level of Brucella isolates with MALDI-TOF-MS

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    Lista Florigio

    2011-12-01

    Full Text Available Abstract Background The genus Brucella contains highly infectious species that are classified as biological threat agents. The timely detection and identification of the microorganism involved is essential for an effective response not only to biological warfare attacks but also to natural outbreaks. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS is a rapid method for the analysis of biological samples. The advantages of this method, compared to conventional techniques, are rapidity, cost-effectiveness, accuracy and suitability for the high-throughput identification of bacteria. Discrepancies between taxonomy and genetic relatedness on the species and biovar level complicate the development of detection and identification assays. Results In this study, the accurate identification of Brucella species using MALDI-TOF-MS was achieved by constructing a Brucella reference library based on multilocus variable-number tandem repeat analysis (MLVA data. By comparing MS-spectra from Brucella species against a custom-made MALDI-TOF-MS reference library, MALDI-TOF-MS could be used as a rapid identification method for Brucella species. In this way, 99.3% of the 152 isolates tested were identified at the species level, and B. suis biovar 1 and 2 were identified at the level of their biovar. This result demonstrates that for Brucella, even minimal genomic differences between these serovars translate to specific proteomic differences. Conclusions MALDI-TOF-MS can be developed into a fast and reliable identification method for genetically highly related species when potential taxonomic and genetic inconsistencies are taken into consideration during the generation of the reference library.

  4. Spatial distribution and risk factors of Brucellosis in Iberian wild ungulates

    Directory of Open Access Journals (Sweden)

    de la Fuente José

    2010-03-01

    Full Text Available Abstract Background The role of wildlife as a brucellosis reservoir for humans and domestic livestock remains to be properly established. The aim of this work was to determine the aetiology, apparent prevalence, spatial distribution and risk factors for brucellosis transmission in several Iberian wild ungulates. Methods A multi-species indirect immunosorbent assay (iELISA using Brucella S-LPS antigen was developed. In several regions having brucellosis in livestock, individual serum samples were taken between 1999 and 2009 from 2,579 wild bovids, 6,448 wild cervids and4,454 Eurasian wild boar (Sus scrofa, and tested to assess brucellosis apparent prevalence. Strains isolated from wild boar were characterized to identify the presence of markers shared with the strains isolated from domestic pigs. Results Mean apparent prevalence below 0.5% was identified in chamois (Rupicapra pyrenaica, Iberian wild goat (Capra pyrenaica, and red deer (Cervus elaphus. Roe deer (Capreolus capreolus, fallow deer (Dama dama, mouflon (Ovis aries and Barbary sheep (Ammotragus lervia tested were seronegative. Only one red deer and one Iberian wild goat resulted positive in culture, isolating B. abortus biovar 1 and B. melitensis biovar 1, respectively. Apparent prevalence in wild boar ranged from 25% to 46% in the different regions studied, with the highest figures detected in South-Central Spain. The probability of wild boar being positive in the iELISA was also affected by age, age-by-sex interaction, sampling month, and the density of outdoor domestic pigs. A total of 104 bacterial isolates were obtained from wild boar, being all identified as B. suis biovar 2. DNA polymorphisms were similar to those found in domestic pigs. Conclusions In conclusion, brucellosis in wild boar is widespread in the Iberian Peninsula, thus representing an important threat for domestic pigs. By contrast, wild ruminants were not identified as a significant brucellosis reservoir for

  5. Characterisation of the genetic diversity of Brucella by multilocus sequencing

    Directory of Open Access Journals (Sweden)

    MacMillan Alastair P

    2007-04-01

    Full Text Available Abstract Background Brucella species include economically important zoonotic pathogens that can infect a wide range of animals. There are currently six classically recognised species of Brucella although, as yet unnamed, isolates from various marine mammal species have been reported. In order to investigate genetic relationships within the group and identify potential diagnostic markers we have sequenced multiple genetic loci from a large sample of Brucella isolates representing the known diversity of the genus. Results Nine discrete genomic loci corresponding to 4,396 bp of sequence were examined from 160 Brucella isolates. By assigning each distinct allele at a locus an arbitrary numerical designation the population was found to represent 27 distinct sequence types (STs. Diversity at each locus ranged from 1.03–2.45% while overall genetic diversity equated to 1.5%. Most loci examined represent housekeeping gene loci and, in all but one case, the ratio of non-synonymous to synonymous change was substantially Brucella species, B. abortus, B. melitensis, B. ovis and B. neotomae correspond to well-separated clusters. With the exception of biovar 5, B. suis isolates cluster together, although they form a more diverse group than other classical species with a number of distinct STs corresponding to the remaining four biovars. B. canis isolates are located on the same branch very closely related to, but distinguishable from, B. suis biovar 3 and 4 isolates. Marine mammal isolates represent a distinct, though rather weakly supported, cluster within which individual STs display one of three clear host preferences. Conclusion The sequence database provides a powerful dataset for addressing ongoing controversies in Brucella taxonomy and a tool for unambiguously placing atypical, phenotypically discordant or newly emerging Brucella isolates. Furthermore, by using the phylogenetic backbone described here, robust and rationally selected markers for use in

  6. Virulence factors and genetic variability of Staphylococcus aureus strains isolated from raw sheep's milk cheese.

    Science.gov (United States)

    Spanu, Vincenzo; Spanu, Carlo; Virdis, Salvatore; Cossu, Francesca; Scarano, Christian; De Santis, Enrico Pietro Luigi

    2012-02-01

    Contamination of dairy products with Staphylococcus aureus can be of animal or human origin. The host pathogen relationship is an important factor determining genetic polymorphism of the strains and their potential virulence. The aim of the present study was to carry out an extensive characterization of virulence factors and to study the genetic variability of S. aureus strains isolated from raw ewe's milk cheese. A total of 100 S. aureus strains isolated from cheese samples produced in 10 artisan cheese factories were analyzed for the presence of enterotoxins (sea-see) and enterotoxins-like genes (seh, sek, sel, sem, seo, sep), leukocidins, exfoliatins, haemolysins, toxic shock syndrome toxin 1 (TSST-1) and the accessory gene regulator alleles (agr). Strains were also typed using pulsed-field gel electrophoresis (PFGE). AMOVA analysis carried out on PFGE and PCR data showed that the major component explaining genetic distance between strains was the dairy of origin. Of the total isolates 81% had a pathogenicity profile ascribable to "animal" biovar while 16% could be related to "human" biovar. The biovar allowed to estimate the most likely origin of the contamination. Minimum inhibitory concentrations (MICs) of nine antimicrobial agents and the presence of the corresponding genes coding for antibiotic resistance was also investigated. 18 strains carrying blaZ gene showed resistance to ampicillin and penicillin and 6 strains carrying tetM gene were resistant to tetracycline. The presence of mecA gene and methicillin resistance, typical of strains of human origin, was never detected. The results obtained in the present study confirm that S. aureus contamination in artisan cheese production is mainly of animal origin. Copyright © 2011. Published by Elsevier B.V.

  7. [Standard algorithm of molecular typing of Yersinia pestis strains].

    Science.gov (United States)

    Eroshenko, G A; Odinokov, G N; Kukleva, L M; Pavlova, A I; Krasnov, Ia M; Shavina, N Iu; Guseva, N P; Vinogradova, N A; Kutyrev, V V

    2012-01-01

    Development of the standard algorithm of molecular typing of Yersinia pestis that ensures establishing of subspecies, biovar and focus membership of the studied isolate. Determination of the characteristic strain genotypes of plague infectious agent of main and nonmain subspecies from various natural foci of plague of the Russian Federation and the near abroad. Genotyping of 192 natural Y. pestis strains of main and nonmain subspecies was performed by using PCR methods, multilocus sequencing and multilocus analysis of variable tandem repeat number. A standard algorithm of molecular typing of plague infectious agent including several stages of Yersinia pestis differentiation by membership: in main and nonmain subspecies, various biovars of the main subspecies, specific subspecies; natural foci and geographic territories was developed. The algorithm is based on 3 typing methods--PCR, multilocus sequence typing and multilocus analysis of variable tandem repeat number using standard DNA targets--life support genes (terC, ilvN, inv, glpD, napA, rhaS and araC) and 7 loci of variable tandem repeats (ms01, ms04, ms06, ms07, ms46, ms62, ms70). The effectiveness of the developed algorithm is shown on the large number of natural Y. pestis strains. Characteristic sequence types of Y. pestis strains of various subspecies and biovars as well as MLVA7 genotypes of strains from natural foci of plague of the Russian Federation and the near abroad were established. The application of the developed algorithm will increase the effectiveness of epidemiologic monitoring of plague infectious agent, and analysis of epidemics and outbreaks of plague with establishing the source of origin of the strain and routes of introduction of the infection.

  8. Evaluation and selection of tandem repeat loci for a Brucella MLVA typing assay

    Directory of Open Access Journals (Sweden)

    Denoeud France

    2006-02-01

    Full Text Available Abstract Background The classification of Brucella into species and biovars relies on phenotypic characteristics and sometimes raises difficulties in the interpretation of the results due to an absence of standardization of the typing reagents. In addition, the resolution of this biotyping is moderate and requires the manipulation of the living agent. More efficient DNA-based methods are needed, and this work explores the suitability of multiple locus variable number tandem repeats analysis (MLVA for both typing and species identification. Results Eighty tandem repeat loci predicted to be polymorphic by genome sequence analysis of three available Brucella genome sequences were tested for polymorphism by genotyping 21 Brucella strains (18 reference strains representing the six 'classical' species and all biovars as well as 3 marine mammal strains currently recognized as members of two new species. The MLVA data efficiently cluster the strains as expected according to their species and biovar. For practical use, a subset of 15 loci preserving this clustering was selected and applied to the typing of 236 isolates. Using this MLVA-15 assay, the clusters generated correspond to the classical biotyping scheme of Brucella spp. The 15 markers have been divided into two groups, one comprising 8 user-friendly minisatellite markers with a good species identification capability (panel 1 and another complementary group of 7 microsatellite markers with higher discriminatory power (panel 2. Conclusion The MLVA-15 assay can be applied to large collections of Brucella strains with automated or manual procedures, and can be proposed as a complement, or even a substitute, of classical biotyping methods. This is facilitated by the fact that MLVA is based on non-infectious material (DNA whereas the biotyping procedure itself requires the manipulation of the living agent. The data produced can be queried on a dedicated MLVA web service site.

  9. Molecular epidemiology of Corynebacterium pseudotuberculosis isolated from horses in California.

    Science.gov (United States)

    Haas, Dionei J; Dorneles, Elaine M S; Spier, Sharon J; Carroll, Scott P; Edman, Judy; Azevedo, Vasco A; Heinemann, Marcos B; Lage, Andrey P

    2017-04-01

    Corynebacterium pseudotuberculosis biovar Equi is an important pathogen of horses. It is increasing in frequency in the United States, and is responsible for various clinical forms of infection, including external abscesses, internal abscesses of the abdominal or thoracic cavities, and ulcerative lymphangitis. The host/pathogen factors dictating the form or severity of infection are currently unknown. Our recent investigations have shown that genotyping C. pseudotuberculosis isolates using enterobacterial repetitive intergenic consensus (ERIC)-PCR is useful for understanding the evolutionary genetics of the species as well for molecular epidemiology studies. The aims of the present study were to assess (i) the genetic diversity of C. pseudotuberculosis strains isolated from horses in California, United States and (ii) the epidemiologic relationships among isolates. One hundred and seven C. pseudotuberculosis biovar Equi isolates from ninety-five horses, and two C. pseudotuberculosis biovar Ovis strains, C. pseudotuberculosis ATCC 19410 T type strain and C. pseudotuberculosis 1002 vaccine strain, were fingerprinted using the ERIC 1+2-PCR. C. pseudotuberculosis isolated from horses showed a high genetic diversity, clustering in twenty-seven genotypes with a diversity index of 0.91. Minimal spanning tree showed four major clonal complexes with a pattern of temporal clustering. Strains isolated from the same horse showed identical ERIC 1+2-PCR genotype, with the exception of two strains isolated from the same animal that showed distinct genotypes, suggesting a co-infection. We found no strong genetic signals related to clinical form (including internal versus external infections). However, temporal clustering of genotypes was observed. Copyright © 2016. Published by Elsevier B.V.

  10. Ureaplasma: Current perspectives

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    P Kokkayil

    2015-01-01

    Full Text Available Ureaplasma species are the most prevalent genital Mycoplasma isolated from the urogenital tract of both men and women. Ureaplasma has 14 known serotypes and is divided into two biovars- Ureaplasma parvum and Ureaplasma urealyticum. The organism has several genes coding for surface proteins, the most important being the gene encoding the Multiple Banded Antigen (MBA. The C-terminal domain of MBA is antigenic and elicits a host antibody response. Other virulence factors include phospholipases A and C, IgA protease and urease. Besides genital tract infections and infertility, Ureaplasma is also associated with adverse pregnancy outcomes and diseases in the newborn (chronic lung disease and retinopathy of prematurity. Infection produces cytokines in the amniotic fluid which initiates preterm labour. They have also been reported from renal stone and suppurative arthritis. Genital infections have also been reported with an increasing frequency in HIV-infected patients. Ureaplasma may be a candidate ′co factor′ in the pathogenesis of AIDS. Culture and polymerase chain reaction (PCR are the mainstay of diagnosis. Commercial assays are available with improved turnaround time. Micro broth dilution is routinely used to test antimicrobial susceptibility of isolates. The organisms are tested against azithromycin, josamycin, ofloxacin and doxycycline. Resistance to macrolides, tetracyclines and fluoroquinolones have been reported. The susceptibility pattern also varies among the biovars with biovar 2 maintaining higher sensitivity rates. Prompt diagnosis and initiation of appropriate antibiotic therapy is essential to prevent long term complications of Ureaplasma infections. After surveying PubMed literature using the terms ′Ureaplasma′, ′Ureaplasma urealyticum′ and ′Ureaplasma parvum′, relevant literature were selected to provide a concise review on the recent developments.

  11. Ureaplasma: current perspectives.

    Science.gov (United States)

    Kokkayil, P; Dhawan, B

    2015-01-01

    Ureaplasma species are the most prevalent genital Mycoplasma isolated from the urogenital tract of both men and women. Ureaplasma has 14 known serotypes and is divided into two biovars- Ureaplasma parvum and Ureaplasma urealyticum. The organism has several genes coding for surface proteins, the most important being the gene encoding the Multiple Banded Antigen (MBA). The C-terminal domain of MBA is antigenic and elicits a host antibody response. Other virulence factors include phospholipases A and C, IgA protease and urease. Besides genital tract infections and infertility, Ureaplasma is also associated with adverse pregnancy outcomes and diseases in the newborn (chronic lung disease and retinopathy of prematurity). Infection produces cytokines in the amniotic fluid which initiates preterm labour. They have also been reported from renal stone and suppurative arthritis. Genital infections have also been reported with an increasing frequency in HIV-infected patients. Ureaplasma may be a candidate 'co factor' in the pathogenesis of AIDS. Culture and polymerase chain reaction (PCR) are the mainstay of diagnosis. Commercial assays are available with improved turnaround time. Micro broth dilution is routinely used to test antimicrobial susceptibility of isolates. The organisms are tested against azithromycin, josamycin, ofloxacin and doxycycline. Resistance to macrolides, tetracyclines and fluoroquinolones have been reported. The susceptibility pattern also varies among the biovars with biovar 2 maintaining higher sensitivity rates. Prompt diagnosis and initiation of appropriate antibiotic therapy is essential to prevent long term complications of Ureaplasma infections. After surveying PubMed literature using the terms 'Ureaplasma', 'Ureaplasma urealyticum' and 'Ureaplasma parvum', relevant literature were selected to provide a concise review on the recent developments.

  12. Emended descriptions of the genus Micrococcus, Micrococcus luteus (Cohn 1872) and Micrococcus lylae (Kloos et al. 1974).

    Science.gov (United States)

    Wieser, Monika; Denner, Ewald B M; Kämpfer, Peter; Schumann, Peter; Tindall, Brian; Steiner, Ulrike; Vybiral, Dietmar; Lubitz, Werner; Maszenan, A M; Patel, B K C; Seviour, Robert J; Radax, Christian; Busse, Hans-Jürgen

    2002-03-01

    Nine yellow-pigmented, spherical bacterial strains isolated from a medieval wall painting (strain D7), from indoor air (strains 3, 6, 7, 13C2, 38, 83 and 118) and from an activated-sludge plant (strain Ballarat) were classified by a polyphasic approach. Analyses of the 16S rRNA gene sequences of three representatives (strains D7, 118 and Ballarat) indicated that they all belong to the genus Micrococcus. The three isolates shared the highest sequence similarities with Micrococcus luteus DSM 20030T (97.9-98%), Micrococcus antarcticus AS 1.2372T (97.9-98.3%) and Micrococcus lylae DSM 20315T (97.5-97.9%). DNA-DNA reassociation studies clearly demonstrated that all nine isolates belong to the species M. luteus. However, neither their chemotaxonomic features nor their physiological and biochemical properties were consistent with those of M. luteus DSM 20030T. In contrast to M. luteus DSM 20030T, all isolates investigated possessed MK-8(H2) as the major respiratory quinone, and strain Ballarat had an A4alpha peptidoglycan type. On the basis of analyses of their Fourier transform-infrared spectroscopy spectra, isolates D7, 3, 6, 7, 13C2, 38, 83 and 118 could be grouped into a single cluster separate from M. luteus DSM 20030T, strain Ballarat and M. lylae DSM 20315T. In addition, all these isolates could be distinguished from M. luteus DSM 20030T by their ability to assimilate D-maltose, D-trehalose, DL-3-hydroxybutyrate, DL-lactate, pyruvate and L-histidine and to hydrolyse casein. Strains D7, 3, 6, 7, 13C2, 38, 83 and 118 differed from both M. luteus DSM 20030T and strain Ballarat by their ability to assimilate acetate, L-phenylalanine, L-serine and phenylacetate. Furthermore, REP-PCR fingerprinting yielded one common band for these strains, whereas this band was not observed for M. luteus DSM 20030T, strain Ballarat or M. lylae DSM 20315T. On the basis of these data, the species M. luteus can be divided into three biovars that are distinguished by several chemotaxonomic

  13. Characteristics of β-lactamases and their genes (blaA and blaB in Yersinia intermedia and Y. frederiksenii

    Directory of Open Access Journals (Sweden)

    Sharma Sachin

    2007-04-01

    Full Text Available Abstract Background The presence of β-lactamases in Y. enterocolitica has been reported to vary with serovars, biovars and geographical origin of the isolates. An understanding of the β-lactamases in other related species is important for an overall perception of antibiotic resistance in yersiniae. The objective of this work was to study the characteristics of β-lactamases and their genes in strains of Y. intermedia and Y. frederiksenii, isolated from clinical and non-clinical sources in India. Results The enzymes, Bla-A (a constitutive class A penicillinase and Bla-B (an inducible class C cephalosporinase were found to be present in all the clinical and non-clinical strains of Y. intermedia and Y. frederiksenii by double disc diffusion method. The results showed differential expression of Bla-A as indicated by presence/absence of synergy whereas expression of Bla-B was quite consistent. The presence of these enzymes was also reflected in the high minimum inhibitory concentrations, MIC50 (126–1024 mg/L and MIC90 (256–1024 mg/L of β-lactam antibiotics against these species. Restriction fragment length polymorphism (RFLP revealed heterogeneity in both blaA and blaB genes of Y. intermedia and Y. frederiksenii. The blaA gene of Y. intermedia shared significant sequence identity (87–96% with blaA of Y. enterocolitica biovars 1A, 1B and 4. The sequence identity of blaA of Y. frederiksenii with these biovars was 77–79%. The sequence identity of blaB gene of Y. intermedia and Y. frederiksenii was more (85% with that of Y. enterocolitica biovars 1A, 1B and 2 compared to other species viz., Y. bercovieri, Y. aldovae and Y. ruckeri. Isoelectric focusing data further revealed that both Y. intermedia and Y. frederiksenii produced Bla-A (pI 8.7 and "Bla-B like" (pI 5.5–7.1 enzymes. Conclusion Both Y. intermedia and Y. frederiksenii showed presence of blaA and blaB genes and unequivocal expression of the two β-lactamases. Limited heterogeneity

  14. Tips and tricks for the assembly of a Corynebacterium pseudotuberculosis genome using a semiconductor sequencer

    DEFF Research Database (Denmark)

    Ramos, Rommel Thiago Jucá; Carneiro, Adriana Ribeiro; Soares, Siomar de Castro

    2013-01-01

    that enable reference-based assembly, such as the one used in the present study, Corynebacterium pseudotuberculosis biovar equi, which causes high economic losses in the US equine industry. The quality treatment strategy incorporated into the assembly pipeline enabled a 16-fold greater use of the sequencing...... was validated by comparative genomics with other species of the genus Corynebacterium. The present study presents a modus operandi that enables a greater and better use of data obtained from semiconductor sequencing for obtaining the complete genome from a prokaryotic microorganism, C. pseudotuberculosis, which...

  15. ORF Alignment: NC_003143 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_003143 gi|16124118 >1pp9A 5 435 24 460 3e-27 ... ref|YP_072312.1| putative insulina...y3837 ... [Yersinia pestis KIM] gb|AAS63519.1| putative insulinase ... family protease [Yersin...ia pestis biovar Medievalis str. ... 91001] ref|NP_994642.1| putative insulina...ypothetical protein [Yersinia pestis ... KIM] emb|CAC93452.1| putative insulinase family protease ... ... ... [Yersinia pestis CO92] ref|NP_407431.1| putative ... insulinase fa

  16. ORF Alignment: NC_004088 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_004088 gi|22127708 >1pp9A 5 435 24 460 3e-27 ... ref|YP_072312.1| putative insulina...y3837 ... [Yersinia pestis KIM] gb|AAS63519.1| putative insulinase ... family protease [Yersin...ia pestis biovar Medievalis str. ... 91001] ref|NP_994642.1| putative insulina...ypothetical protein [Yersinia pestis ... KIM] emb|CAC93452.1| putative insulinase family protease ... ... ... [Yersinia pestis CO92] ref|NP_407431.1| putative ... insulinase fa

  17. ORF Alignment: NC_006155 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_006155 gi|51598121 >1pp9A 5 435 24 460 3e-27 ... ref|YP_072312.1| putative insulina...y3837 ... [Yersinia pestis KIM] gb|AAS63519.1| putative insulinase ... family protease [Yersin...ia pestis biovar Medievalis str. ... 91001] ref|NP_994642.1| putative insulina...ypothetical protein [Yersinia pestis ... KIM] emb|CAC93452.1| putative insulinase family protease ... ... ... [Yersinia pestis CO92] ref|NP_407431.1| putative ... insulinase fa

  18. ORF Alignment: NC_005810 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_005810 gi|45443103 >1pp9A 5 435 24 460 3e-27 ... ref|YP_072312.1| putative insulina...y3837 ... [Yersinia pestis KIM] gb|AAS63519.1| putative insulinase ... family protease [Yersin...ia pestis biovar Medievalis str. ... 91001] ref|NP_994642.1| putative insulina...ypothetical protein [Yersinia pestis ... KIM] emb|CAC93452.1| putative insulinase family protease ... ... ... [Yersinia pestis CO92] ref|NP_407431.1| putative ... insulinase fa

  19. ORF Alignment: NC_005810 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_005810 gi|45441027 >1p71A 1 90 1 91 3e-24 ... ref|YP_069949.1| integration host fa...ctor beta-subunit [Yersinia pseudotuberculosis ... IP 32953] ref|NP_670080.1| integration host factor ... ... (IHF), beta subunit [Yersinia pestis KIM] ... gb|AAS61443.1| integration host factor beta-su...bunit ... [Yersinia pestis biovar Medievalis str. 91001] ... ref|NP_992566.1| integration...ration host factor (IHF), beta ... subunit [Yersinia pestis KIM] ref|NP_404986.1| ... integratio

  20. Genome-scale reconstruction of the metabolic network in Yersinia pestis CO92

    Science.gov (United States)

    Navid, Ali; Almaas, Eivind

    2007-03-01

    The gram-negative bacterium Yersinia pestis is the causative agent of bubonic plague. Using publicly available genomic, biochemical and physiological data, we have developed a constraint-based flux balance model of metabolism in the CO92 strain (biovar Orientalis) of this organism. The metabolic reactions were appropriately compartmentalized, and the model accounts for the exchange of metabolites, as well as the import of nutrients and export of waste products. We have characterized the metabolic capabilities and phenotypes of this organism, after comparing the model predictions with available experimental observations to evaluate accuracy and completeness. We have also begun preliminary studies into how cellular metabolism affects virulence.

  1. Mycobacteria of clinical interest

    International Nuclear Information System (INIS)

    Casal, M.

    1986-01-01

    This book is based upon a symposium on mycobacteria of clinical interest. Due to the multidisciplinary participation of, among others, microbiologists, clinicians, immunologists and epidemiologists, a very wide and thorough presentation of the present state of clinical research in this field is ensured. Topics of particular interest included in this volume were the new antimicrobial agents active against mycobacteria; new therapeutic possibilities; a system of rapid diagnosis of tuberculosis and mycobacteriosis; mycobacteriosis in AIDS; progress in immunopathology of tuberculosis and leprosy; progress in bacteriology and vaccination in leprosy; progress in immunological diagnosis and new epidemiological biovars of M. tuberculosis. (Auth.)

  2. ORF Alignment: NC_004310 [GENIUS II[Archive

    Lifescience Database Archive (English)

    Full Text Available NC_004310 gi|23502485 >1bxwA 4 170 45 261 5e-15 ... gb|AAN30527.1| outer membrane protein Omp3...1 [Brucella suis 1330] gb|AAS84575.1| ... Omp31b [Brucella cetaceae] gb|AAS84574.1| Omp31b ... ... ... [Brucella cetaceae] gb|AAS84573.1| Omp31b [Brucella ... cetaceae] gb|AAS84572.1| Omp31b [Br...ucella pinnipediae] ... gb|AAS84571.1| Omp31b [Brucella melitensis biovar ... ... Neotomae] ref|NP_698612.1| outer membrane protein Omp31 ... [Brucella suis 1330] ...

  3. A case of pharyngeal diphtheria in Germany, June 2015.

    Science.gov (United States)

    Berger, A; Meinel, D M; Schaffer, A; Ziegler, R; Pitteroff, J; Konrad, R; Sing, Andreas

    2016-10-01

    In June 2015, a 45-year-old man suffering from acute necrotic tonsillitis and throat phlegmon was hospitalized in Nuremberg, Germany. After emergency surgery the patient was initially treated with antibiotics. A throat swab grew a toxigenic Corynebacterium diphtheriae biovar mitis strain. The patient's vaccination status was not documented and the patient was tested serologically for anti-diphtheria antibodies showing no protective immunity. Extensive control investigations were performed by the local health department showing no likely source of his infection. No secondary cases were found and the patient completely recovered.

  4. [Content of salicylic and jasmonic acids in pea roots (Pisum sativum L.) at the initial stage of symbiotic or pathogenic interaction with bacteria of the family Rhizobiaceae].

    Science.gov (United States)

    Rudikovskaya, E G; Akimova, G P; Rudikovskii, A V; Katysheva, N B; Dudareva, L V

    2017-01-01

    A change in the contents of endogenous salicylic and jasmonic acids in the roots of the host plant at the preinfectious stage of interaction with symbiotic (Rhizobium leguminosarum) and pathogenic (Agrobacterium rizogenes) bacteria belonging for to the family Rhizobiaceae was studied. It was found that the jasmonic acid content increased 1.5–2 times 5 min after inoculation with these bacterial species. It was shown that dynamics of the change in the JA and SA contents depends on the type of infection. Thus, the JA content decreased in the case of pathogenesis, while the SA content increased. At the same time, an increased JA content was observed during symbiosis. The observed regularities could indicate the presence of different strategies of hormonal regulation for interaction with symbiotic and pathogenic bacteria belonging to the family Rhizobiaceae in peas plants.

  5. Response of Phaseolus vulgaris cv. Mantequilla to inoculation of Rhizobium native strains of the Ecuador in greenhouse native races of the Ecuador

    Directory of Open Access Journals (Sweden)

    Klever Iván Granda Mora

    2016-10-01

    Full Text Available The development of biofertilizers based on Rhizobium strains for common bean (Phaseolus vulgaris L. requires studies about bacterial interaction with target cultivars. For these reason, the aim of this paper was to determine the effect of the inoculation of Rhizobium native strains, isolated from southern Ecuador, on P. vulgaris cultivar Mantequilla. An assay was performed in greenhouse. It were evaluated the parameters of nodulation, biomass, nitrogen fixation and efficiency of the symbiosis. All inoculated strains were able to nodulate bean seedlings. The total number of nodules, nodular biomass and plant biomass, were favourably affected by inoculation of Rhizobium strains. The best results were obtained with R. mesoamericanum NAM1, R. leguminosarum COL 6 and R. etli PIN 1 strains. The experimental evidences shows the potential of native strains of Rhizobium for it use as biofertilizers because they are able to raise the rates of nitrogen fixation in common bean in southern Ecuador.

  6. Symbiotic Activity of Pea (Pisum sativum after Application of Nod Factors under Field Conditions

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    Anna Siczek

    2014-04-01

    Full Text Available Growth and symbiotic activity of legumes are mediated by Nod factors (LCO, lipo-chitooligosaccharides. To assess the effects of application of Nod factors on symbiotic activity and yield of pea, a two-year field experiment was conducted on a Haplic Luvisol developed from loess. Nod factors were isolated from Rhizobium leguminosarum bv. viciae strain GR09. Pea seeds were treated with the Nod factors (10−11 M or water (control before planting. Symbiotic activity was evaluated by measurements of nitrogenase activity (acetylene reduction assay, nodule number and mass, and top growth by shoot mass, leaf area, and seed and protein yield. Nod factors generally improved pea yield and nitrogenase activity in the relatively dry growing season 2012, but not in the wet growing season in 2013 due to different weather conditions.

  7. Structural Insight Into the Role of Mutual Polymorphism and Conservatism in the Contact Zone of the NFR5-K1 Heterodimer With the Nod Factor.

    Science.gov (United States)

    Igolkina, A A; Porozov, Yu B; Chizhevskaya, E P; Andronov, E E

    2018-01-01

    Sandwich-like docking configurations of the heterodimeric complex of NFR5 and K1 Vicia sativa receptor-like kinases together with the putative ligand, Nod factor (NF) of Rhizobium leguminosarum bv. viciae , were modeled and two of the most probable configurations were assessed through the analysis of the mutual polymorphisms and conservatism. We carried out this analysis based on the hypothesis that in a contact zone of two docked components (proteins or ligands) the population polymorphism or conservatism is mutual, i.e., the variation in one component has a reflected variation in the other component. The population material of 30 wild-growing V. sativa (leaf pieces) was collected from a large field (uncultivated for the past 25-years) and pooled; form this pool, 100 randomly selected cloned fragments of NFR5 gene and 100 of K1 gene were sequenced by the Sanger method. Congruence between population trees of NFR5 and K1 haplotypes allowed us to select two respective haplotypes, build their 3D structures, and perform protein-protein docking. In a separate simulation, the protein-ligand docking between NFR5 and NF was carried out. We merged the results of the two docking experiments and extracted NFR5-NF-K1 complexes, in which NF was located within the cavity between two receptors. Molecular dynamics simulations indicated two out of six complexes as stable. Regions of mutual polymorphism in the contact zone of one complex overlapped with known NF structural variations produced by R. leguminosarum bv. viciae . A total of 74% of the contact zone of another complex contained mutually polymorphic and conservative areas. Common traits of the obtained two stable structures allowed us to hypothesize the functional role of three-domain structure of plant LysM-RLKs in their heteromers.

  8. Some long term effects on land application of sewage sludge on soil fertility

    International Nuclear Information System (INIS)

    McGrath, S.P.; Chaudri, A.M.; Giller, K.E.

    1997-01-01

    Metals may affect the growth of plants, soil microbial activity, and soil fertility in the long term. Less is known of the adverse long-term effects of metals on soil microorganisms than on crop yields, since the effects of metals added to soils in sewage sludge are difficult to assess, and few long-term experiments exist. This paper reviews evidence from controlled long-term field experiments with sewage sludges in the UK, Sweden, Germany and the USA. Adverse effects on microbial activity and populations of cyanobacteria (blue-green algae), Rhizobium leguminosarum bv. trifolii, mycorrhiza, and total microbial biomass have been detected, in some cases below the European Community's maximum allowable concentration limits for metals in sludge-treated soils. For example, N 2 -fixation by free- living heterotrophic bacteria was found to be inhibited at concentrations (mg kg -1 ) of 127 Zn, 37 Cu, 21 Ni, 3.4 Cd, 52 Cr and 71 Pb. Fixation by free-living cyanobacteria was reduced by 50% at concentrations (mg kg -1 ) of 114 Zn, 33 Cu, 17 Ni, 2.9 Cd, 80 Cr and 40 Pb. Numbers of Rhizobium Leguminosarum bv. trifolii were decreased by several orders of magnitude at metal concentrations (mg kg -1 ) of 130-200 Zn, 27-48 Cu, 11-15 Ni, and 0.8-1.0 Cd. Important factors influencing the severity of toxicity are soil texture and pH; higher pH and clay and organic C contents decrease metal toxicity considerably. The evidence presented in this review of long-term field experiments suggests that adverse effects on microbial parameters occur at modest concentrations of metals in soils. It is concluded that prevention of adverse effects on soil microbial processes, and ultimately on soil fertility, should be a factor that influences soil-protection legislation. (author)

  9. Structural Insight Into the Role of Mutual Polymorphism and Conservatism in the Contact Zone of the NFR5–K1 Heterodimer With the Nod Factor

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    A. A. Igolkina

    2018-04-01

    Full Text Available Sandwich-like docking configurations of the heterodimeric complex of NFR5 and K1 Vicia sativa receptor-like kinases together with the putative ligand, Nod factor (NF of Rhizobium leguminosarum bv. viciae, were modeled and two of the most probable configurations were assessed through the analysis of the mutual polymorphisms and conservatism. We carried out this analysis based on the hypothesis that in a contact zone of two docked components (proteins or ligands the population polymorphism or conservatism is mutual, i.e., the variation in one component has a reflected variation in the other component. The population material of 30 wild-growing V. sativa (leaf pieces was collected from a large field (uncultivated for the past 25-years and pooled; form this pool, 100 randomly selected cloned fragments of NFR5 gene and 100 of K1 gene were sequenced by the Sanger method. Congruence between population trees of NFR5 and K1 haplotypes allowed us to select two respective haplotypes, build their 3D structures, and perform protein–protein docking. In a separate simulation, the protein-ligand docking between NFR5 and NF was carried out. We merged the results of the two docking experiments and extracted NFR5–NF–K1 complexes, in which NF was located within the cavity between two receptors. Molecular dynamics simulations indicated two out of six complexes as stable. Regions of mutual polymorphism in the contact zone of one complex overlapped with known NF structural variations produced by R. leguminosarum bv. viciae. A total of 74% of the contact zone of another complex contained mutually polymorphic and conservative areas. Common traits of the obtained two stable structures allowed us to hypothesize the functional role of three-domain structure of plant LysM-RLKs in their heteromers.

  10. [Methods for the detection of Agrobacterium from plant, soil and water samples].

    Science.gov (United States)

    Alippi, Adriana M; López, Ana C; Balatti, Pedro A

    2011-01-01

    The genus Agrobacterium includes phytopathogenic bacteria that induce the development of root crown galls and/or aerial galls at the base of the stem or hairy roots on more than 600 species of plants belonging to 90 dicotyledonous families and non-pathogenic species. These bacteria being natural soil inhabitants are particularly difficult to eradicate, which is a problem in nurseries where more than 80% of infections occur. Since early detection is crucial to avoid the inadvertent spread of the disease, the aim of this work was to develop sensitive and precise identification techniques by using a set of semi-selective and differential culture media in combination with a specific PCR to amplify a partial sequence derived from the virC operon, as well as a multiplex PCR on the basis of 23SrDNA sequences, and biological assays to identify and differentiate species and biovars of Agrobacterium obtained either from soil, water or plant samples. The combination of the different assays allowed us to reduce the number of false positive and negative results from bacteria isolated from any of the three types of samples. Therefore, the combination of multiplex PCR, specific PCR, isolations in semi-selective D1, D1-M and YEM-RCT media combined with bioassays on cut leaves of Kalanchoe and seedlings of California Wonder pepper cultivar constitute an accurate tool to detect species and biovars of Agrobacterium for diagnostic purposes.

  11. Evaluation of Cholera Toxin Expression in Acidic, Alkaline and Neutral Conditions

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    Narges Rahimi

    2015-02-01

    Full Text Available Background: Cholera is a severe disease which is caused by Vibrio cholerae and it is typically transmitted by either contaminated food or water particularly in developing countries. The most important virulence factor of this bacterium is an enterotoxin called cholera toxin which is a protein complex secreted by the Vibrio cholerae. Objectives: In this project, we determined the production of cholera toxin at different pH values. Materials and Methods: Two standard strain of Vibrio cholerae O1 biovar EL Tor N16961 and Vibrio cholerae O1 biovar Classic ATCC 14035 were used. After overnight cultivation of both the strains the total mRNA extracted and converted to total cDNA. Results: By Relative Real-Time PCR analysis the most cholera toxin production in classical and El Tor strains was at pH 8.5 and 8, respectively. Conclusions: Therefore, We may conclude that use of acidic diet will help in reduction of cholera toxin production.

  12. Vibrios among patients of good socioeconomic conditions during the cholera epidemic in Recife, Brazil Vibriões coléricos e não coléricos entre pacientes de boas condições sódo-econômicas durante a epidemia de coléra no Recife, Brasil

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    Vera Magalhães

    1993-08-01

    Full Text Available Between March and July, 1992, we screened for Vibrio all fecal samples submitted for bacteriologic diagnosis at a private clinical laboratory in Recife. Of 1435 cultures examined only 1 (0.07% was positive for V.cholerae 01, biovar Eltor, serovar Inaba, but 17 (1.2% yielded non-cholera Vibrio (V.cholerae non-01; V.fluvialis; V.furnissii, V.parahaemolyticus and Vibrio spp. Thus, V.cholerae 01, differently of other enteropathogenic vibrios, spared individuals of good socioeconomic conditions even during the cholera epidemic, which made hundreds of victims in the neighboring slums.Entre março e julho de 1992, pesquisou-se Vibrio em todos os espécimes fecais enviados para diagnóstico bacteriológico a um laboratório clínico privado do Recife. De 1435 culturas examinadas apenas 1 (0.07% foi positiva para V.cholerae 01, biovar Eltor, sorovar Inaba, porém 17 (1,2% forneceram outras espécies de Vibrio (V.cholerae nao-01; V.fluvialis; V.furnissii; V.parahaemolyticus e Vibrio spp. Portanto, V.cholerae 01, diferentemente de outros vibriões entero patogênicos, poupou indivíduos de boas condições sócio-econômicas, mesmo durante uma epidemia de cólera que atingiu centenas de pessoas nas favelas vizinhas.

  13. Swine brucellosis: current perspectives

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    Olsen SC

    2016-12-01

    Full Text Available SC Olsen, FM Tatum Infectious Bacterial Diseases of Livestock Research Unit, National Animal Disease Center, Agricultural Research Service, United States Department of Agriculture, Ames, IA, USA Abstract: Brucella suis is a significant zoonotic species that is present in domestic livestock and wildlife in many countries worldwide. Transmission from animal reservoirs is the source of human infection as human-to-human transmission is very rare. Although swine brucellosis causes economic losses in domestic livestock, preventing human infection is the primary reason for its emphasis in disease control programs. Although disease prevalence varies worldwide, in areas outside of Europe, swine brucellosis is predominantly caused by B. suis biovars 1 and 3. In Europe, swine are predominantly infected with biovar 2 which is much less pathogenic in humans. In many areas worldwide, feral or wild populations of swine are important reservoir hosts. Like other Brucella spp. in their natural host, B. suis has developed mechanisms to survive in an intracellular environment and evade immune detection. Limitations in sensitivity and specificity of current diagnostics require use at a herd level, rather for individual animals. There is currently no commercial vaccine approved for preventing brucellosis in swine. Although not feasible in all situations, whole-herd depopulation is the most effective regulatory mechanism to control swine brucellosis. Keywords: livestock, transmission, pathogenicity, vaccine, host, infection

  14. Ralstonia solanacearum – a New Threat to Potato Production in Serbia

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    Svetlana Milijašević-Marčić

    2013-12-01

    Full Text Available A survey of ware potatoes (a total of 1127 samples from localities in Serbia during two consecutive years resulted in detection and identification of R. solanacearum in 17 tuber samples. The monitoring detected the causal agent of bacterial wilt and brown rot of potato in three districts of Vojvodina province. In 2011, the infection by R. solanacearum was confirmed in 7 samples of ware potato tubers (varieties – Saturna, Pirol, Hermes, Panda in West Bačka and South Bačka Districts. In 2012, the infection by R. solanacearum was confirmed in 10 potato tuber samples (Lady Claire, Desiree, Panda, Red Fantasy and Vineta varieties from two districts: South Bačka and Central Banat. Bacterial strains obtained from positive samples were identified as R. solanacearum biovar 2 using PCR/RFLP analysis, pathogenicity test on tomato transplants, and nutritional, enzymatic and biovar determination tests. To our best knowledge, these are the only findings of R. solanacearum infection in ware potatoes in Serbia. R. solanacearum was not detected in tomato or any other host plant tested in this study. Furthermore, the bacterium was not found in any of the water samples tested, including those originating from areas in which the bacterium was found in ware potato samples.

  15. Anti-Brucella Antibodies in Moose (Alces alces gigas), Muskoxen (Ovibos moschatus), and Plains Bison (Bison bison bison) in Alaska, USA.

    Science.gov (United States)

    Nymo, Ingebjørg Helena; Beckmen, Kimberlee; Godfroid, Jacques

    2016-01-01

    We used an indirect enzyme-linked immunosorbent assay (iELISA) and the rose bengal test (RBT) to test for anti-Brucella antibodies in moose (Alces alces gigas), muskoxen (Ovibos moschatus), and plains bison (Bison bison bison) from various game management units (GMUs) in Alaska, US, sampled from 1982 to 2010. A portion of the sera had previously been tested with the standard plate test (SPT), the buffered Brucella antigen (BBA) card test, and the card test (CARD). No antibody-positive plains bison were identified. Anti-Brucella antibodies were detected in moose (iELISA, n=4/87; RBT, n=4/87; SPT, n=4/5; BBA, n=4/4) from GMU 23 captured in 1992, 1993, and 1995 and in muskoxen (iELISA, n=4/52; RBT, n=4/52; CARD, n=4/35) from GMUs 26A and 26B captured in 2004, 2006, and 2007. A negative effect of infection on the health of individuals of these species is probable. The presence of antibody-positive animals from 1992 to 2007 suggests presence of brucellae over time. The antibody-positive animals were found in northern Alaska, an area with a historically higher prevalence of Brucella-positive caribou, and a spillover of Brucella suis biovar 4 from caribou may have occurred. Brucella suis biovar 4 causes human brucellosis, and transmission from consumption of moose and muskoxen is possible.

  16. Pitfalls in the diagnosis and management of inguinal lymphogranuloma venereum: important lessons from a case series.

    Science.gov (United States)

    Oud, Emerentiana Veronica; de Vrieze, Nynke Hesselina Neeltje; de Meij, Arjan; de Vries, Henry John C

    2014-06-01

    Current lymphogranuloma venereum (LGV) guidelines mainly focus on anorectal infections. Inguinal LGV infections have been rare in the current epidemic among men who have sex with men (MSM), but might require a different approach not yet recommended in current guidelines for the treatment and diagnosis of LGV. We describe 4 inguinal LGV cases. Three MSM developed inguinal LGV infection several weeks after a previous consultation, of which two had received azithromycin after being notified for LGV. Three failed the recommended 21 days doxycycline treatment. These inguinal LGV cases highlight 3 pitfalls in the current standard management of LGV: (1) Urethral chlamydia infections in MSM can be caused by LGV biovars that in contrast to non-LGV biovars require prolonged antibiotic therapy. (2) The recommended one gram azithromycin contact treatment seems insufficient to prevent established infections. (3) Inguinal LGV may require prolonged courses of doxycycline, exceeding the currently advised 21 days regimen. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  17. Evaluation of the HOOF-Print assay for typing Brucella abortus strains isolated from cattle in the United States: results with four performance criteria

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    Ewalt Darla R

    2005-06-01

    Full Text Available Abstract Background A fundamental question that arises during epidemiological investigations of bacterial disease outbreaks is whether the outbreak strain is genetically related to a proposed index strain. Highly discriminating genetic markers for characterizing bacterial strains can help in clarifying the genetic relationships among strains. Under the auspices of the European Society of Clinical Microbiology and Infectious Diseases, the European Study Group for Epidemiological Markers (ESGEM established guidelines for evaluating the performance of typing systems based of a number of criteria. Recently, HOOF-Print genotype analysis, a new method for typing Brucella abortus strains based on hypervariability at eight tandem repeat loci, was described. This paper evaluates the HOOF-Print assay by four of the criteria set out by the ESGEM: typeability, reproducibility, power of discrimination, and concordance with other typing methods. Results The HOOF-Print Assay was evaluated with a test population composed of 97 unrelated field isolates and 6 common laboratory strains of B. abortus. Both typeability and reproducibility of the assay were excellent. Allele diversity and frequency varied widely among the eight loci, ranging from 1 to 13 alleles. The power of discrimination, measured by the Hunter-Gaston discrimination index (HGDI, varied by locus ranging from 0 to 0.89, where a maximal value of 1.0 indicates discrimination of all strains. The HGDI values calculated for subgroups sorted by biovar were similar to the values determined for the whole population. None of the individual loci achieved the recommended HGDI threshold of 0.95, but the HGDI of the composite profiles was 0.99 (93 unique genotypes from 97 field strains evaluated, well above the recommended threshold. By comparison, the HGDI value for biovar typing was 0.61 in a test population biased with disproportionate numbers of the less common biovars. Cluster analysis based on HOOF

  18. Ação de tinturas e óleos essenciais de plantas medicinais sobre o crestamento bacteriano comum do feijoeiro e na produção de proteínas de indução de resistência

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    Sandra Cristina Vigo

    2009-12-01

    Full Text Available A exploração da atividade biológica de compostos secundários presentes nas tinturas ou em óleos essenciais de plantas podem representar, ao lado da indução de resistência, mais uma forma potencial de controle de doenças em plantas cultivadas. O presente trabalho objetivou avaliar o potencial de tinturas de Lippia alba, Lippia sidoides, Mikania glomerata, Equisetum sp. e Hedera helix e óleos essenciais de Rosmarinus officinalis e Cinnamomum zeylanicum nas atividades in vitro, in vivo e na produção de proteínas na indução de resistência, em plantas de feijão vagem cultivar Bragança. Os resultados obtidos demonstraram que as tinturas de L. alba e L. sidoides e os óleos essenciais (R. officinalis e C. zeylanicum apresentaram atividade in vitro aos isolados de Xanthomonas axonopodis pv. phaseoli. Todas as tinturas ensaiadas apresentaram menores valores do progresso da doença (AACPD, em relação à testemunha, merecendo destaque a tintura de L. alba, que estavam correlacionadas com os maiores teores de polifenoloxidase, peroxidase e proteínas solúveis totais, evidenciando uma possível indução de resistência. Os óleos essenciais não apresentaram diferença na AACPD e nem na indução de proteínas.Additionally to resistance inducers, the exploitation of secondary compounds biological activity present in plants alcohol extracts or essential oils could potential way to control diseases in cultivated plants. This aimed to evaluate the potential of Lippia alba, Lippia sidoides, Mikania glomerata, Equisetum sp. and Hedera helix alcohol extracts and, Rosmarinus officinalis and Cinnamomum zeylanicum essential oils on in vitro and in vivo activity, and protein production on resistance induction in snap beans Bragança cultivar. Results showed in vitro activity against Xanthomonas axonopodis pv. phaseoli for L. alba and L. sidoides extracts, and essential oils. Although all alcohol extracts have showed the lowest area under the

  19. Evaluation of pyraclostrobin and acibenzolar-S-methyl on common bacterial blight of snap beanAvaliação de pyraclostrobin e acibenzolar-S-methyl sobre o crestamento bacteriano comum do feijão-vagem

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    Renata de Cássia Camara

    2012-04-01

    Full Text Available Assays were done under greenhouse conditions in order to evaluate the effect of pyraclostrobin (0.0375, 0.0750 and 0.150 mL.L-1 and acibenzolar-S-methyl (ASM (0.025 g.L-1 in common bacterial blight, using leaves of snap beans cultivar Bragança. These chemicals were sprayed at three different times: five days before; five days before + five days after; and five days after leaf inoculation with an isolate of Xanthomonas axonopodis pv. phaseoli. They were determinate the levels of polyphenoloxidase, peroxidase and total soluble proteins on inoculated and non-inoculated leaves of snap beans sprayed with pyraclostrobin (0.075 g.L-1 and ASM (0.025 g.L-1. All concentration of pyraclostrobin and ASM reduced the area under the disease progress curve (AUDPC on leaves of snap beans, and the least AUDPC value was observed when this products were sprayed five days before + five days after inoculation. Higher levels of polyphenoloxidase, peroxidase and the total soluble proteins were observed on leaves sprayed with pyraclostrobin or ASM.Ensaios foram conduzidos sob condições de casa-de-vegetação para avaliar o efeito de pyraclostrobin (0,0375; 0.0750 e 0,150 mL.L-1 e acibenzolar-S-methyl (ASM (0.025 g.L-1 sobre o crestamento bacteriano comum, em folhas de feijão-vagem cultivar Bragança. Os produtos foram pulverizados em três diferentes períodos: cinco dias antes, cinco dias antes + cinco dias após e cinco dias após a inoculação dos folíolos com um isolado de Xanthomonas axonopodis pv. phaseoli. Foram determinados os teores de polifenoloxidase, peroxidase e proteínas solúveis totais em folhas inoculadas e não-inoculadas de feijão-vagem pulverizadas com pyraclostrobin (0,075 g.L-1 e ASM (0,025 g.L-1. Todas as concentrações de pyraclostrobin e ASM reduziram os valores da área abaixo da curva do progresso da doença (AUPDC e o menor valor da AUPDC foi observado para a aplicação dos produtos cinco dias antes + cinco dias após a inocula

  20. A rapid minor groove binder PCR method for distinguishing the vaccine strain Brucella abortus 104M.

    Science.gov (United States)

    Nan, Wenlong; Qin, Lide; Wang, Yong; Zhang, Yueyong; Tan, Pengfei; Chen, Yuqi; Mao, Kairong; Chen, Yiping

    2018-01-24

    Brucellosis is a widespread zoonotic disease caused by Gram-negative Brucella bacteria. Immunisation with attenuated vaccine is an effective method of prevention, but it can interfere with diagnosis. Live, attenuated Brucella abortus strain 104M has been used for the prevention of human brucellosis in China since 1965. However, at present, no fast and reliable method exists that can distinguish this strain from field strains. Single nucleotide polymorphism (SNP)-based assays offer a new approach for such discrimination. SNP-based minor groove binder (MGB) and Cycleave assays have been used for rapid identification of four Brucella vaccine strains (B. abortus strains S19, A19 and RB51, and B. melitensis Rev1). The main objective of this study was to develop a PCR assay for rapid and specific detection of strain 104M. We developed a SNP-based MGB PCR assay that could successfully distinguish strain 104M from 18 representative strains of Brucella (B. abortus biovars 1, 2, 3, 4, 5, 6, 7 and 9, B. melitensis biovars 1, 2 and 3, B. suis biovars 1, 2, 3 and 4, B. canis, B. neotomae, and B. ovis), four Brucella vaccine strains (A19, S19, S2, M5), and 55 Brucella clinical field strains. The assay gave a negative reaction with four non-Brucella species (Escherichia coli, Pasteurella multocida, Streptococcus suis and Pseudomonas aeruginosa). The minimum sensitivity of the assay, evaluated using 10-fold dilutions of chromosomal DNA, was 220 fg for the 104M strain and 76 fg for the single non-104M Brucella strain tested (B. abortus A19). The assay was also reproducible (intra- and inter-assay coefficients of variation = 0.006-0.022 and 0.012-0.044, respectively). A SNP-based MGB PCR assay was developed that could straightforwardly and unambiguously distinguish B. abortus vaccine strain 104M from non-104M Brucella strains. Compared to the classical isolation and identification approaches of bacteriology, this real-time PCR assay has substantial advantages in terms of

  1. Effects of rhizobial bacteria on K, Ca and Na concentration of wheat (Triticum aestivum L. in saline soils

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    S homayoon

    2016-05-01

    Full Text Available Introduction Soil salinity is one of the major agricultural problems and it is limiting crop productivity in many parts of the cultivated areas all over the world. Saline soils are differentiated by the presence of great ratios of Na/Ca, Na/K, Ca2+, Mg2+, and Cl/NO3 (Gratan & Catherine, 1993 and high levels of neutral salts in the surface layers, which are resulting from the capillary action (Al-Falih, 2002. Osmotic stress occurs when soluble salts increase in the soils and then results in specific ion toxicity (Agarwal & Ahmad, 2010. Therefore, one of the most important side effects of salinity is nutritional disorders. High concentration of NaCl in the root medium usually reduces nutrients uptake and affects the transportation of potassium and calcium ions in plant. (Gratan & Catherine, 1993 reported that the salinity of soils changes ionic strength of the substrate and it can influence mineral nutrient uptake and translocation. Salinity also changes the mineral nutrient availability and disrupts the mineral relations of plants. Hence, the main purpose of this research is to evaluate the effects of rhizobial bacteria inoculation on K, Ca and Na concentration of wheat (Triticum aestivum L. in saline soils. Material and methods Soil sample was collected from Astan Ghodse Razavi farm, Mashhad Iran, and then was dried and passed through a 12-mesh (approximately 2 mm screen. Soil sample was divided into three parts and then was placed into three containers. Each container was watered by a different proportion of saline water (EC= 10 dS.m-1. Salinity of soils was regularly monitored until three salinities (2, 6 and 10 dS.m-1 came out. Then, a completely randomized design with a factorial arrangement was carried out in a greenhouse condition. The experimental factors included four levels of inoculation (Sinorhizobium meliloti, Bradyrhizobium japonicum and Rhizobium leguminosarum and control and three levels of soil salinity (2, 6 and 10 dS.m-1 with

  2. SEROLOGICAL DIAGNOSIS OF MODERN CHOLERA USING LIPOSOMAL ENTEROTOXIC DIAGNOSTICUM IN COMPLEMENT FIXATION TEST

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    I. V. Savelyeva

    2013-01-01

    Full Text Available Abstract. The possibility of serological diagnosis of cholera using cholera enterotoxic diagnostics kit in complement fixation test to detect anti-enterotoxic antibodies in sera of patients with cholera caused by hybrid variants of the El Tor biovar has been demonstrated. In patients with mild course of cholera anti-enterotoxic antibodies were detected in titres 1:50 and 1:200 in paired sera obtained on the 7th and 14th days of disease, respectively (fourfold titre increase. In patients with the course of medium severity 32-fold titre increase was recorded from the titre 1:100 in serum obtained on the fifth day of disease till the titre 1:3200 — on the twelfth day of disease. Antibodies titers reached 1:1600 and 1:800 were revealed in two medium course patients (adult and infant of 10 months on the sixth day of disease.

  3. Prevalence and resistance to antibiotics for Aeromonas species from retail fish in Malaysia.

    Science.gov (United States)

    Radu, Son; Ahmad, Noorlis; Ling, Foo Hooi; Reezal, Abdul

    2003-03-25

    A total of 87 market fish samples representing five types of fish were evaluated for the presence of Aeromonas spp. Of the samples examined, 69%, 55%, 11.5% and 2.3% harbored Aeromonas spp., A. veronii biovar sobria, A. hydrophila and A. caviae, respectively. The 60 isolated Aeromonas spp. strains were further examined for hemolytic activity, resistance to antimicrobial agents and presence of plasmids. Hemolytic activity varied widely among the isolated strains. Though all the isolates demonstrated resistance to three or more of the antibiotics tested, all were susceptible to ceptazidime. Thirty-four (56.7%) of the sixty isolates harbored plasmids, with sizes ranging from 2.3 to 15.7 kb. These results indicate that hemolytic, multiple antibiotic resistant and genetically diverse aeromonads are easily recovered from fish in this region.

  4. Persistent anthrax as a major driver of wildlife mortality in a tropical rainforest.

    Science.gov (United States)

    Hoffmann, Constanze; Zimmermann, Fee; Biek, Roman; Kuehl, Hjalmar; Nowak, Kathrin; Mundry, Roger; Agbor, Anthony; Angedakin, Samuel; Arandjelovic, Mimi; Blankenburg, Anja; Brazolla, Gregory; Corogenes, Katherine; Couacy-Hymann, Emmanuel; Deschner, Tobias; Dieguez, Paula; Dierks, Karsten; Düx, Ariane; Dupke, Susann; Eshuis, Henk; Formenty, Pierre; Yuh, Yisa Ginath; Goedmakers, Annemarie; Gogarten, Jan F; Granjon, Anne-Céline; McGraw, Scott; Grunow, Roland; Hart, John; Jones, Sorrel; Junker, Jessica; Kiang, John; Langergraber, Kevin; Lapuente, Juan; Lee, Kevin; Leendertz, Siv Aina; Léguillon, Floraine; Leinert, Vera; Löhrich, Therese; Marrocoli, Sergio; Mätz-Rensing, Kerstin; Meier, Amelia; Merkel, Kevin; Metzger, Sonja; Murai, Mizuki; Niedorf, Svenja; De Nys, Hélène; Sachse, Andreas; van Schijndel, Joost; Thiesen, Ulla; Ton, Els; Wu, Doris; Wieler, Lothar H; Boesch, Christophe; Klee, Silke R; Wittig, Roman M; Calvignac-Spencer, Sébastien; Leendertz, Fabian H

    2017-08-02

    Anthrax is a globally important animal disease and zoonosis. Despite this, our current knowledge of anthrax ecology is largely limited to arid ecosystems, where outbreaks are most commonly reported. Here we show that the dynamics of an anthrax-causing agent, Bacillus cereus biovar anthracis, in a tropical rainforest have severe consequences for local wildlife communities. Using data and samples collected over three decades, we show that rainforest anthrax is a persistent and widespread cause of death for a broad range of mammalian hosts. We predict that this pathogen will accelerate the decline and possibly result in the extirpation of local chimpanzee (Pan troglodytes verus) populations. We present the epidemiology of a cryptic pathogen and show that its presence has important implications for conservation.

  5. Molecular typing of isolates obtained from aborted foetuses in Brucella-free Holstein dairy cattle herd after immunisation with Brucella abortus RB51 vaccine in Egypt.

    Science.gov (United States)

    Wareth, Gamal; Melzer, Falk; Böttcher, Denny; El-Diasty, Mohamed; El-Beskawy, Mohamed; Rasheed, Nesma; Schmoock, Gernot; Roesler, Uwe; Sprague, Lisa D; Neubauer, Heinrich

    2016-12-01

    Bovine brucellosis is endemic in Egypt in spite of application of surveillance and control measures. An increase of abortions was reported in a Holstein dairy cattle herd with 600 animals in Damietta governorate in Egypt after immunisation with Brucella (B.) abortus RB51 vaccine. Twenty one (10.6%) of 197 vaccinated cows aborted after 3 months. All aborted cows had been tested seronegative for brucellosis in the past 3 years. B. abortus was isolated from four foetuses. Conventional biochemical and bacteriological identification and polymerase chain reaction (PCR) confirmed two B. abortus biovar (bv.) 1 smooth and two B. abortus rough strains. None of the B. abortus isolates were identified as RB51. Genotyping analysis by multiple locus of variable number tandem repeats analysis based on 16 markers (MLVA-16) revealed two different profiles with low genetic diversity. B. abortus bv1 was introduced in the herd and caused abortions. Copyright © 2016 Elsevier B.V. All rights reserved.

  6. Lactic Acid Bacteria from Kefir Increase Cytotoxicity of Natural Killer Cells to Tumor Cells.

    Science.gov (United States)

    Yamane, Takuya; Sakamoto, Tatsuji; Nakagaki, Takenori; Nakano, Yoshihisa

    2018-03-27

    The Japanese fermented beverage, homemade kefir, contains six lactic acid bacteria: Lactococcus. lactis subsp. Lactis , Lactococcus . lactis subsp. Cremoris , Lactococcus. Lactis subsp. Lactis biovar diacetylactis , Lactobacillus plantarum , Leuconostoc meseuteroides subsp. Cremoris and Lactobacillus casei . In this study, we found that a mixture of the six lactic acid bacteria from kefir increased the cytotoxicity of human natural killer KHYG-1 cells to human chronic myelogenous leukemia K562 cells and colorectal tumor HCT116 cells. Furthermore, levels of mRNA expression and secretion of IFN-γ (interferon gamma) increased in KHYG-1 cells that had been treated with the six lactic acid bacteria mixture from kefir. The results suggest that the six lactic acid bacteria mixture from kefir has strong effects on natural immunity and tumor cell cytotoxicity.

  7. Persistent anthrax as a major driver of wildlife mortality in a tropical rainforest

    Science.gov (United States)

    Hoffmann, Constanze; Zimmermann, Fee; Biek, Roman; Kuehl, Hjalmar; Nowak, Kathrin; Mundry, Roger; Agbor, Anthony; Angedakin, Samuel; Arandjelovic, Mimi; Blankenburg, Anja; Brazolla, Gregory; Corogenes, Katherine; Couacy-Hymann, Emmanuel; Deschner, Tobias; Dieguez, Paula; Dierks, Karsten; Düx, Ariane; Dupke, Susann; Eshuis, Henk; Formenty, Pierre; Yuh, Yisa Ginath; Goedmakers, Annemarie; Gogarten, Jan F.; Granjon, Anne-Céline; McGraw, Scott; Grunow, Roland; Hart, John; Jones, Sorrel; Junker, Jessica; Kiang, John; Langergraber, Kevin; Lapuente, Juan; Lee, Kevin; Leendertz, Siv Aina; Léguillon, Floraine; Leinert, Vera; Löhrich, Therese; Marrocoli, Sergio; Mätz-Rensing, Kerstin; Meier, Amelia; Merkel, Kevin; Metzger, Sonja; Murai, Mizuki; Niedorf, Svenja; de Nys, Hélène; Sachse, Andreas; van Schijndel, Joost; Thiesen, Ulla; Ton, Els; Wu, Doris; Wieler, Lothar H.; Boesch, Christophe; Klee, Silke R.; Wittig, Roman M.; Calvignac-Spencer, Sébastien; Leendertz, Fabian H.

    2017-08-01

    Anthrax is a globally important animal disease and zoonosis. Despite this, our current knowledge of anthrax ecology is largely limited to arid ecosystems, where outbreaks are most commonly reported. Here we show that the dynamics of an anthrax-causing agent, Bacillus cereus biovar anthracis, in a tropical rainforest have severe consequences for local wildlife communities. Using data and samples collected over three decades, we show that rainforest anthrax is a persistent and widespread cause of death for a broad range of mammalian hosts. We predict that this pathogen will accelerate the decline and possibly result in the extirpation of local chimpanzee (Pan troglodytes verus) populations. We present the epidemiology of a cryptic pathogen and show that its presence has important implications for conservation.

  8. Lactic Acid Bacteria from Kefir Increase Cytotoxicity of Natural Killer Cells to Tumor Cells

    Directory of Open Access Journals (Sweden)

    Takuya Yamane

    2018-03-01

    Full Text Available The Japanese fermented beverage, homemade kefir, contains six lactic acid bacteria: Lactococcus. lactis subsp. Lactis, Lactococcus. lactis subsp. Cremoris, Lactococcus. Lactis subsp. Lactis biovar diacetylactis, Lactobacillus plantarum, Leuconostoc meseuteroides subsp. Cremoris and Lactobacillus casei. In this study, we found that a mixture of the six lactic acid bacteria from kefir increased the cytotoxicity of human natural killer KHYG-1 cells to human chronic myelogenous leukemia K562 cells and colorectal tumor HCT116 cells. Furthermore, levels of mRNA expression and secretion of IFN-γ (interferon gamma increased in KHYG-1 cells that had been treated with the six lactic acid bacteria mixture from kefir. The results suggest that the six lactic acid bacteria mixture from kefir has strong effects on natural immunity and tumor cell cytotoxicity.

  9. Monitoring of phytopathogenic Ralstonia solanacearum cells using green fluorescent protein-expressing plasmid derived from bacteriophage phiRSS1.

    Science.gov (United States)

    Kawasaki, Takeru; Satsuma, Hideki; Fujie, Makoto; Usami, Shoji; Yamada, Takashi

    2007-12-01

    A green fluorescent protein (GFP)-expressing plasmid was constructed from a filamentous bacteriophage phiRSS1 that infects the phytopathogen Ralstonia solanacearum. This plasmid designated as pRSS12 (4.7 kbp in size) consists of an approximately 2248 bp region of the phiRSS1 RF DNA, including ORF1-ORF3 and the intergenic region (IG), and a Km cassette in addition to the GFP gene. It was easily introduced by electroporation and stably maintained even without selective pressure in strains of R. solanacearum of different races and biovars. Strong green fluorescence emitted from pRSS12-transformed bacterial cells was easily monitored in tomato tissues (stem, petiole, and root) after infection as well as from soil samples. These results suggest that pRSS12 can serve as an easy-to-use GFP-tagging tool for any given strain of R. solanacearum in cytological as well as field studies.

  10. Seroprevalence of Brucella spp. in Cattle, Molecular Characterization in Milk, and the Analysis of Associated Risk Factors with Seroprevalence in Humans, Egypt.

    Science.gov (United States)

    El-Diasty, Mohamed M; Ahmed, Heba A; Sayour, Ashraf E; El Hofy, Fatma I; Tahoun, Asmaa B M B; Shafik, Saleh M

    2016-12-01

    The objective of the present study was to estimate the seroprevalence of Brucella spp. in humans and cattle at Sharkia Governorate, Egypt. In addition, identification of Brucella spp. in milk samples by PCR and culture with the evaluation of the risk factors associated with Brucella spp. seroprevalence in humans were carried out. Overall, the seroprevalence of Brucella antibodies in the examined cattle was 23.8%, while in human participants it was 21%. The examination of 205 milk samples using PCR revealed that 6.3% were positive for B. abortus biovar 1 and the results were confirmed by culture methods. Multivariate logistic regression revealed that consumption of unpasteurized dairy products, occupational contact with animals, and knowledge about the disease are risk factors associated with infection in humans. This study documented the endemic status of brucellosis in Egypt. Hygienic measures and increased awareness about the disease are recommended to minimize the spread of infection from animals to humans.

  11. Cell-mediated immune responses differentiate infections with Brucella suis from Yersinia enterocolitica serotype O : 9 in pigs

    DEFF Research Database (Denmark)

    Riber, Ulla; Jungersen, Gregers

    2007-01-01

    Due to almost identical lipopolysaccharide (LPS) O-antigens, infections with Yersinia enterocolitica serotype 0:9 (YeO:9) cause false positive serological reactions (FPSR) in tests for Brucella and thus cause problems in National Brucella surveillance programs. As LPS are strong inducers...... of antibody responses it was hypothesized that cell-mediated immune responses to non-LPS antigens of the two bacteria can be used to separate immune responses to these two biologically very different infections. Following subclinical experimental infections with Brucella suis biovar 2, high interferon......-gamma (IFN-gamma) assay responses with a commercial Brucella melitensis antigen preparation (Brucellergene OCB) preceded the development of antibodies. High IFN-gamma responses in the seven B. suis inoculated pigs with serological evidence of infection were consistent throughout a 20-week postinoculation...

  12. Genomic comparisons of Brucella spp. and closely related bacteria using base compositional and proteome based methods

    DEFF Research Database (Denmark)

    Bohlin, Jon; Snipen, Lars; Cloeckaert, Axel

    2010-01-01

    BACKGROUND: Classification of bacteria within the genus Brucella has been difficult due in part to considerable genomic homogeneity between the different species and biovars, in spite of clear differences in phenotypes. Therefore, many different methods have been used to assess Brucella taxonomy....... In the current work, we examine 32 sequenced genomes from genus Brucella representing the six classical species, as well as more recently described species, using bioinformatical methods. Comparisons were made at the level of genomic DNA using oligonucleotide based methods (Markov chain based genomic signatures...... between the oligonucleotide based methods used. Whilst the Markov chain based genomic signatures grouped the different species in genus Brucella according to host preference, the codon and amino acid frequencies based methods reflected small differences between the Brucella species. Only minor differences...

  13. Diphtheria outbreak in Maranhão, Brazil: microbiological, clinical and epidemiological aspects.

    Science.gov (United States)

    Santos, L S; Sant'anna, L O; Ramos, J N; Ladeira, E M; Stavracakis-Peixoto, R; Borges, L L G; Santos, C S; Napoleão, F; Camello, T C F; Pereira, G A; Hirata, R; Vieira, V V; Cosme, L M S S; Sabbadini, P S; Mattos-Guaraldi, A L

    2015-03-01

    We describe microbiological, clinical and epidemiological aspects of a diphtheria outbreak that occurred in Maranhão, Brazil. The majority of the 27 confirmed cases occurred in partially (n = 16) or completely (n = 10) immunized children (n = 26). Clinical signs and characteristic symptoms of diphtheria such as cervical lymphadenopathy and pseudomembrane formation were absent in 48% and 7% of the cases, respectively. Complications such as paralysis of lower limbs were observed. Three cases resulted in death, two of them in completely immunized children. Microbiological analysis identified the isolates as Corynebacterium diphtheriae biovar intermedius with a predominant PFGE type. Most of them were toxigenic and some showed a decrease in penicillin G susceptibility. In conclusion, diphtheria remains endemic in Brazil. Health professionals need to be aware of the possibility of atypical cases of C. diphtheriae infection, including pharyngitis without pseudomembrane formation.

  14. Improvements in the Flavour of Soy Cheese

    Directory of Open Access Journals (Sweden)

    Naveed Ahmad

    2008-01-01

    Full Text Available A review of biochemical and technological similarities and dissimilarities between soy cheese and Cheddar cheese is presented to provide guidelines for the improvements in the flavour of soy cheese. Processing technology as well as the final product of soy cheese have many similarities with Cheddar in terms of appearance, texture, mouth feel, chemical nature, biochemical processes, etc. Soy protein has many useful amino acids like Asp, Ile, Leu, Met, Phe, Trp, Tyr, Val, etc., which are precursors of flavouring compounds and the right choice of microbial cultures is necessary to benefit from them. Using low levels of sodium chloride, without the use of ethanol, and introducing new milk cheese starter and non-starter cultures like Lactococcus lactis ssp. lactis (formerly L. lactis ssp. lactis biovar. diacetylactis, Lactobacillus helveticus, Lactobacillus casei, Streptococcus lactis var. maltigenes and Lactococcus lactis ssp. cremoris that enhance flavour will be helpful to improve the flavour of soy cheese.

  15. Characterisation of Yersinia Secretion Apparatus--Pathogenicity Island (Ysa-PI) of Yersinia enterocolitica 1B/O8 in Poland: an Idle Ysa is a Specific Hallmark of the Epidemic Sensu Stricto Strain.

    Science.gov (United States)

    Wołkowicz, Tomasz; Zacharczuk, Katarzyna; Rokosz-Chudziak, Natalia; Rastawicki, Waldemar; Gierczyński, Rafał

    2015-01-01

    Yersinia secretion apparatus (Ysa), the chromosomal type three secretion system (T3SS) is considered to contribute to virulence of high-pathogenicity Yersina enterocolitica biovar 1B. DNA-sequence of Ysa pathogenicity island was determined for clinical isolate DM0110 of Y enterocolitica 1B/08 with origin in Poland. We found a premature stop-codon in the regulatory gene ysrR (mutation at position 269). Altered ysrR was detected in all tested 78 isolates of Y enterocolitica 1B/O8 collected from clinical samples in Poland from 2004 to 2013. Since aberrations in YsrR are considered to inactivate Ysa, our findings may suggest Ysa is not indispensable for Y enterocolitica 1B/O8 to infect humans.

  16. T4-related bacteriophage LIMEstone isolates for the control of soft rot on potato caused by 'Dickeya solani'.

    Directory of Open Access Journals (Sweden)

    Evelien M Adriaenssens

    Full Text Available The bacterium 'Dickeya solani', an aggressive biovar 3 variant of Dickeya dianthicola, causes rotting and blackleg in potato. To control this pathogen using bacteriophage therapy, we isolated and characterized two closely related and specific bacteriophages, vB_DsoM_LIMEstone1 and vB_DsoM_LIMEstone2. The LIMEstone phages have a T4-related genome organization and share DNA similarity with Salmonella phage ViI. Microbiological and molecular characterization of the phages deemed them suitable and promising for use in phage therapy. The phages reduced disease incidence and severity on potato tubers in laboratory assays. In addition, in a field trial of potato tubers, when infected with 'Dickeya solani', the experimental phage treatment resulted in a higher yield. These results form the basis for the development of a bacteriophage-based biocontrol of potato plants and tubers as an alternative for the use of antibiotics.

  17. Ação antagônica de rizobactérias contra Phytophthora parasitica e p. citrophthora e seu efeito no desenvolvimento de plântulas de citros

    Directory of Open Access Journals (Sweden)

    AMORIM EDNA PEIXOTO DA ROCHA

    2002-01-01

    Full Text Available O antagonismo de Pseudomonas putida biovar A (C1-1B, P. putida biovar B (Santa Bárbara, P. fluorescens (C2-8C e RA2, Bacillus subtilis (OG e RC2 e Flavobacterium sp. (CIS/NA contra Phytophthora parasitica e P. citrophthora , agentes da podridão radicular dos citros, foi avaliado através da inibição do crescimento micelial (cultura pareada e redução na percentagem de infecção da doença em mudas de citros (tratamento de sementes com rizobactérias. Na seleção preliminar, 33 isolados bacterianos foram testados. Sementes de citros pré-germinadas foram tratadas por imersão nas suspensões das bactérias (10(9 ufc/ml, e plantadas em tubetes contendo solo natural infestado com o fitopatógeno (50 ml de suspensão/ kg de solo. A avaliação da percentagem de infecção foi efetuada após 15 dias. In vitro, os isolados bacterianos RC2, OG, CIS/NA e C1-1B foram os mais ativos inibidores do crescimento micelial de Phytophthora. Em condições de casa de vegetação, todos os isolados proporcionaram redução na percentagem de infecção da doença em todos os ensaios realizados. Promoção de crescimento de plantas foi verificada pela inoculação de plântulas com as linhagens OG, RC2, CiS/Na e C1-1B.

  18. Identification and Genetic Characterization of Ralstonia solanacearum Species Complex Isolates from Cucurbita maxima in China

    Directory of Open Access Journals (Sweden)

    Xiaoman She

    2017-10-01

    Full Text Available Ralstonia solanacearum species complex is a devastating phytopathogen with an unusually wide host range, and new host plants are continuously being discovered. In June 2016, a new bacterial wilt on Cucurbita maxima was observed in Guangdong province, China. Initially, in the adult plant stage, several leaves of each plant withered suddenly and drooped; the plant then wilted completely, and the color of their vasculature changed to dark brown, ultimately causing the entire plant to die. Creamy-whitish bacterial masses were observed to ooze from crosscut stems of these diseased plants. To develop control strategies for C. maxima bacterial wilt, the causative pathogenic isolates were identified and characterized. Twenty-four bacterial isolates were obtained from diseased C. maxima plants, and 16S rRNA gene sequencing and pathogenicity analysis results indicated that the pathogen of C. maxima bacterial wilt was Ralstonia solanacearum. The results from DNA-based analysis, host range determination and bacteriological identification confirmed that the 24 isolates belonged to R. solanacearum phylotype I, race 1, and eight of these isolates belonged to biovar 3, while 16 belonged to biovar 4. Based on the results of partial egl gene sequence analysis, the 24 isolates clustered into three egl- sequence type groups, sequevars 17, 45, and 56. Sequevar 56 is a new sequevar which is described for the first time in this paper. An assessment of the resistance of 21 pumpkin cultivars revealed that C. moschata cv. Xiangyu1 is resistant to strain RS378, C. moschata cv. Xiangmi is moderately resistant to strain RS378, and 19 other pumpkin cultivars, including four C. maxima cultivars and 15 C. moschata cultivars, are susceptible to strain RS378. To the best of our knowledge, this is the first report of C. maxima bacterial wilt caused by R. solanacearum race 1 in the world. Our results provide valuable information for the further development of control strategies

  19. Isolation and Characterization of Agrobacterium Strains from Soil: A Laboratory Capstone Experience

    Directory of Open Access Journals (Sweden)

    Kim R. Finer

    2016-12-01

    Full Text Available In this investigation, the students’ goal was to isolate and characterize Agrobacterium strains from soil. Following selection and enrichment on 1A-t medium, putative Agrobacterium isolates were characterized by Gram stain reaction and biochemical tests. Isolates were further evaluated using polymerase chain reaction (PCR with different primer sets designed to amplify specific regions of bacterial deoxyribonucleic acid (DNA. Primer sets included AGRH to identify isolates that were members of the Rhizobiaceae, BIOVAR1 primers to identify members of Agrobacterium biovar group I, and a third set, VIRG, to determine presence of virG (only present in pathogenic Agrobacterium strains. During the investigation, students applied previously learned techniques including serial dilution, use of selective/differential media, staining protocols, biochemical analysis, molecular analysis via PCR, and electrophoresis. Students also gained practical experience using photo documentation to record data for an eventual mock journal publication of the capstone laboratory experience. Pre- and post-evaluation of class content knowledge related to the techniques, protocols, and learning objectives of these laboratories revealed significant learning gains in the content areas of Agrobacterium–plant interactions (p ≤ 0.001 and molecular biology (p ≤ 0.01. The capstone journal assignment served as the assessment tool to evaluate mastery and application of laboratory technique, the ability to accurately collect and evaluate data, and critical thinking skills associated with experimental troubleshooting and extrapolation. Analysis of journal reports following the capstone experience showed significant improvement in assignment scores (p ≤ 0.0001 and attainment of capstone experience learning outcomes.

  20. Aeromonas spp. isolated from oysters (Crassostrea rhizophorea from a natural oyster bed, Ceará, Brazil Aeromonas spp. isoladas de ostras (Crassostrea rhizophorea coletadas em um criadouro natural, Ceará, Brazil

    Directory of Open Access Journals (Sweden)

    Norma S. Evangelista-Barreto

    2006-06-01

    Full Text Available Between April and October 2002, thirty fortnightly collections of oysters (Crassostrea rhizophorea from a natural oyster bed at the Cocó River estuary in the Sabiaguaba region (Fortaleza, Ceará, Brazil were carried out, aiming to isolate Aeromonas spp. strains. Oyster samples were submitted to the direct plating (DP and the presence/absence (P/A methods. Aeromonas were identified in 15 (50% samples analyzed by the DP method and in 13 (43% analyzed by the P/A method. A. caviae, A. eucrenophila, A. media, A. sobria, A. trota, A. veronii bv. sobria, A. veronii bv. veronii and Aeromonas sp. were isolated. The predominant species was A. veronii (both biovars, which was identified in 13 (43% samples, followed by A. media in 11 (37% and A. caviae in seven (23%. From the 59 strains identified, 28 (48% presented resistance to at least one of the eight antibiotics tested.Foram realizadas 30 coletas quinzenais, entre abril e outubro de 2002, de ostras (Crassostrea rhizophorea de um criadouro natural, no estuário do rio Cocó (Fortaleza/Ceará/Brasil, objetivando-se isolar cepas de Aeromonas spp. As amostras de ostras foram submetidas aos métodos de plaqueamento direto (PD e presença/ausência (P/A. Foram identificadas Aeromonas em 15 (50% amostras analisadas pelo método PD e em 13 (43% pelo método P/A. Foram isoladas: A. caviae, A. eucrenophila, A. media, A. sobria, A. trota, A. veronii bv. sobria, A. veronii bv. veronii e Aeromonas sp. A espécie predominate foi A. veronii (ambos biovars, identificada em 13 (43% amostras, seguida de A. media em 11 (37% e A. caviae em 7 (23%. Das 59 cepas identificadas, 28 (48% apresentaram resistência a pelo menos um, dos oitos antibióticos testados.

  1. Identification and Genetic Characterization of Ralstonia solanacearum Species Complex Isolates from Cucurbita maxima in China.

    Science.gov (United States)

    She, Xiaoman; Yu, Lin; Lan, Guobing; Tang, Yafei; He, Zifu

    2017-01-01

    Ralstonia solanacearum species complex is a devastating phytopathogen with an unusually wide host range, and new host plants are continuously being discovered. In June 2016, a new bacterial wilt on Cucurbita maxima was observed in Guangdong province, China. Initially, in the adult plant stage, several leaves of each plant withered suddenly and drooped; the plant then wilted completely, and the color of their vasculature changed to dark brown, ultimately causing the entire plant to die. Creamy-whitish bacterial masses were observed to ooze from crosscut stems of these diseased plants. To develop control strategies for C. maxima bacterial wilt, the causative pathogenic isolates were identified and characterized. Twenty-four bacterial isolates were obtained from diseased C. maxima plants, and 16S rRNA gene sequencing and pathogenicity analysis results indicated that the pathogen of C. maxima bacterial wilt was Ralstonia solanacearum . The results from DNA-based analysis, host range determination and bacteriological identification confirmed that the 24 isolates belonged to R. solanacearum phylotype I, race 1, and eight of these isolates belonged to biovar 3, while 16 belonged to biovar 4. Based on the results of partial egl gene sequence analysis, the 24 isolates clustered into three egl- sequence type groups, sequevars 17, 45, and 56. Sequevar 56 is a new sequevar which is described for the first time in this paper. An assessment of the resistance of 21 pumpkin cultivars revealed that C. moschata cv. Xiangyu1 is resistant to strain RS378, C. moschata cv. Xiangmi is moderately resistant to strain RS378, and 19 other pumpkin cultivars, including four C. maxima cultivars and 15 C. moschata cultivars, are susceptible to strain RS378. To the best of our knowledge, this is the first report of C. maxima bacterial wilt caused by R. solanacearum race 1 in the world. Our results provide valuable information for the further development of control strategies for C. maxima wilt

  2. Identification of a single-nucleotide insertion in the promoter region affecting the sodC promoter activity in Brucella neotomae.

    Directory of Open Access Journals (Sweden)

    Dina A Moustafa

    Full Text Available Brucella neotomae is not known to be associated with clinical disease in any host species. Previous research suggested that B. neotomae might not express detectable levels of Cu/Zn superoxide dismutase (SOD, a periplasmic enzyme known to be involved in protecting Brucella from oxidative bactericidal effects of host phagocytes. This study was undertaken to investigate the genetic basis for the disparity in SOD expression in B. neotomae. Our Western blot and SOD enzyme assay analyses indicated that B. neotomae does express SOD, but at a substantially reduced level. Nucleotide sequence analysis of region upstream to the sodC gene identified a single-nucleotide insertion in the potential promoter region. The same single-nucleotide insertion was also detected in the sodC promoter of B. suis strain Thomsen, belonging to biovar 2 in which SOD expression was undetectable previously. Examination of the sodC promoter activities using translational fusion constructs with E. coli β-galactosidase demonstrated that the B. neotomae and B. suis biovar 2 promoters were very weak in driving gene expression. Site-directed mutation studies indicated that the insertion of A in the B. neotomae sodC promoter reduced the promoter activity. Increasing the level of SOD expression in B. neotomae through complementation with B. abortus sodC gene did not alter the bacterial survival in J774A.1 macrophage-like cells and in tissues of BALB/c and C57BL/6 mice. These results for the first time demonstrate the occurrence of a single-nucleotide polymorphism affecting promoter function and gene expression in Brucella.

  3. A Novel PCR Assay for Detecting Brucella abortus and Brucella melitensis.

    Science.gov (United States)

    Alamian, Saeed; Esmaelizad, Majid; Zahraei, Taghi; Etemadi, Afshar; Mohammadi, Mohsen; Afshar, Davoud; Ghaderi, Soheila

    2017-02-01

    Brucellosis is a major zoonotic disease that poses a significant public health threat worldwide. The classical bacteriological detection process used to identify Brucella spp. is difficult and time-consuming. This study aimed to develop a novel molecular assay for detecting brucellosis. All complete sequences of chromosome 1 with 2.1-Mbp lengths were compared among all available Brucella sequences. A unique repeat sequence (URS) locus on chromosome 1 could differentiate Brucella abortus from Brucella melitensis . A primer set was designed to flank the unique locus. A total of 136 lymph nodes and blood samples were evaluated and classified by the URS-polymerase chain reaction (PCR) method in 2013-2014. Biochemical tests and bacteriophage typing as the golden standard indicated that all Brucella spp. isolates were B. melitensis biovar 1 and B. abortus biovar 3. The PCR results were the same as the bacteriological method for detecting Brucella spp. The sensitivity and specificity of the URS-PCR method make it suitable for detecting B. abortus and B. melitensis . Quick detection of B. abortus and B. melitensis can provide the most effective strategies for control of these bacteria. The advantage of this method over other presented methods is that both B. abortus and B. melitensis are detectable in a single test tube. Furthermore, this method covered 100% of all B. melitensis and B. abortus biotypes. The development of this URS-PCR method is the first step toward the development of a novel kit for the molecular identification of B. abortus and B. melitensis .

  4. Whole-genome analyses of speciation events in pathogenic Brucellae

    Energy Technology Data Exchange (ETDEWEB)

    Chain, Patrick S. G. [Lawrence Livermore National Laboratory (LLNL); Comerci, Diego J. [Universidad Nacional de General San Martin; Tolmasky, Marcelo E. [California State University; Larimer, Frank W [ORNL; Malfatti, Stephanie [Lawrence Livermore National Laboratory (LLNL); Vergez, Lisa [Lawrence Livermore National Laboratory (LLNL); Aguero, Fernan [Universidad Nacional de General San Martin; Land, Miriam L [ORNL; Ugalde, Rodolfo A. [Universidad Nacional de General San Martin; Garcia, Emilio [Lawrence Livermore National Laboratory (LLNL)

    2005-12-01

    Despite their high DNA identity and a proposal to group classical Brucella species as biovars of Brucella melitensis, the commonly recognized Brucella species can be distinguished by distinct biochemical and fatty acid characters, as well as by a marked host range (e.g., Brucella suis for swine, B. melitensis for sheep and goats, and Brucella abortus for cattle). Here we present the genome of B. abortus 2308, the virulent prototype biovar 1 strain, and its comparison to the two other human pathogenic Brucella species and to B. abortus field isolate 9-941. The global distribution of pseudogenes, deletions, and insertions supports previous indications that B. abortus and B. melitensis share a common ancestor that diverged from B. suis. With the exception of a dozen genes, the genetic complements of both B. abortus strains are identical, whereas the three species differ in gene content and pseudogenes. The pattern of species-specific gene inactivations affecting transcriptional regulators and outer membrane proteins suggests that these inactivations may play an important role in the establishment of host specificity and may have been a primary driver of speciation in the genus Brucella. Despite being nonmotile, the brucellae contain flagellum gene clusters and display species-specific flagellar gene inactivations, which lead to the putative generation of different versions of flagellum-derived structures and may contribute to differences in host specificity and virulence. Metabolic changes such as the lack of complete metabolic pathways for the synthesis of numerous compounds (e.g., glycogen, biotin, NAD, and choline) are consistent with adaptation of brucellae to an intracellular life-style.

  5. Pathology of brucellosis in bison from Yellowstone National Park

    Science.gov (United States)

    Rhyan, Jack C.; Gidlewski, T.; Roffe, T.J.; Aune, K.; Philo, L.M.; Ewalt, D.R.

    2001-01-01

    Between February 1995 and June 1999, specimens from seven aborted bison (Bison bison) fetuses or stillborn calves and their placentas, two additional placentas, three dead neonates, one 2-wk-old calf, and 35 juvenile and adult female bison from Yellowstone National Park (USA) were submitted for bacteriologic and histopathologic examination. One adult animal with a retained placenta had recently aborted. Serum samples from the 35 juvenile and adult bison were tested for Brucella spp. antibodies. Twenty-six bison, including the cow with the retained placenta, were seropositive, one was suspect, and eight were seronegative. Brucella abortus biovar 1 was isolated from three aborted fetuses and associated placentas, an additional placenta, the 2-wk-old calf, and 11 of the seropositive female bison including the animal that had recently aborted. Brucella abortus biovar 2 was isolated from one additional seropositive adult female bison. Brucella abortus was recovered from numerous tissue sites from the aborted fetuses, placentas and 2-wk-old calf. In the juvenile and adult bison, the organism was more frequently isolated from supramammary (83%), retropharyngeal (67%), and iliac (58%) lymph nodes than from other tissues cultured. Cultures from the seronegative and suspect bison were negative for B. abortus. Lesions in the B. abortus-infected, aborted placentas and fetuses consisted of necropurulent placentitis and mild bronchointerstitial pneumonia. The infected 2-wk-old calf had bronchointerstitial pneumonia, focal splenic infarction, and purulent nephritis. The recently-aborting bison cow had purulent endometritis and necropurulent placentitis. Immunohistochemical staining of tissues from the culture-positive aborted fetuses, placentas, 2-wk-old calf, and recently-aborting cow disclosed large numbers of B. abortus in placental trophoblasts and exudate, and fetal and calf lung. A similar study with the same tissue collection and culture protocol was done using six

  6. Whole-genome analyses of the speciation events in the pathogenic Brucellae

    Energy Technology Data Exchange (ETDEWEB)

    Chain, P; Comerci, D; Tolmasky, M; Larimer, F; Malfatti, S; Vergez, L; Aguero, F; Land, M; Ugalde, R; Garcia, E

    2005-07-14

    Despite their high DNA identity and a proposal to group classical Brucella species as biovars of B. melitensis, the commonly recognized Brucella species can be distinguished by distinct biochemical and fatty acid characters as well as by a marked host range (e.g. B. suis for swine, B. melitensis for sheep and goats, B. abortus for cattle). Here we present the genome of B. abortus 2308, the virulent prototype biovar 1 strain, and its comparison to the two other human pathogenic Brucellae species and to the B. abortus field isolate 9-941. The global distribution of pseudogenes, deletions and insertions support previous indications that B. abortus and B. melitensis share a common ancestor that diverged from B. suis. With the exception of a dozen genes, the genetic complement of both B. abortus strains is identical, whereas the three species differ in gene content and pseudogenes. The pattern of species-specific gene inactivations affecting transcriptional regulators and outer membrane proteins suggest that these inactivations may play an important role in the establishment of host-specificity and may have been a primary driver of speciation in the Brucellae. Despite being non-motile, the Brucellae contain flagellum gene clusters and display species-specific flagellar gene inactivations, which lead to the putative generation of different versions of flagellum-derived structures, and may contribute to differences in host-specificity and virulence. Metabolic changes such as the lack of complete metabolic pathways for the synthesis of numerous compounds (e.g. glycogen, biotin, NAD, and choline) are consistent with adaptation of Brucellae to an intracellular lifestyle.

  7. Phenotypic and molecular characterizations of Yersinia pestis isolates from Kazakhstan and adjacent regions.

    Science.gov (United States)

    Lowell, Jennifer L; Zhansarina, Aigul; Yockey, Brook; Meka-Mechenko, Tatyana; Stybayeva, Gulnaz; Atshabar, Bakyt; Nekrassova, Larissa; Tashmetov, Rinat; Kenghebaeva, Kuralai; Chu, May C; Kosoy, Michael; Antolin, Michael F; Gage, Kenneth L

    2007-01-01

    Recent interest in characterizing infectious agents associated with bioterrorism has resulted in the development of effective pathogen genotyping systems, but this information is rarely combined with phenotypic data. Yersinia pestis, the aetiological agent of plague, has been well defined genotypically on local and worldwide scales using multi-locus variable number tandem repeat analysis (MLVA), with emphasis on evolutionary patterns using old isolate collections from countries where Y. pestis has existed the longest. Worldwide MLVA studies are largely based on isolates that have been in long-term laboratory culture and storage, or on field material from parts of the world where Y. pestis has potentially circulated in nature for thousands of years. Diversity in these isolates suggests that they may no longer represent the wild-type organism phenotypically, including the possibility of altered pathogenicity. This study focused on the phenotypic and genotypic properties of 48 Y. pestis isolates collected from 10 plague foci in and bordering Kazakhstan. Phenotypic characterization was based on diagnostic tests typically performed in reference laboratories working with Y. pestis. MLVA was used to define the genotypic relationships between the central-Asian isolates and a group of North American isolates, and to examine Kazakh Y. pestis diversity according to predefined plague foci and on an intermediate geographical scale. Phenotypic properties revealed that a large portion of this collection lacks one or more plasmids necessary to complete the blocked flea/mammal transmission cycle, has lost Congo red binding capabilities (Pgm-), or both. MLVA analysis classified isolates into previously identified biovars, and in some cases groups of isolates collected within the same plague focus formed a clade. Overall, MLVA did not distinguish unique phylogeographical groups of Y. pestis isolates as defined by plague foci and indicated higher genetic diversity among older biovars.

  8. Clinical manifestations of bacteremia caused by Aeromonas species in southern Taiwan.

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    Hung-Jen Tang

    Full Text Available This study is conducted to investigate the clinical characteristics of patients with bacteremia caused by Aeromonas species.Patients with bacteremia caused by Aeromonas species during the period 2009 to 2013 were identified from a computerized database of a regional hospital in southern Taiwan. The medical records of these patients were retrospectively reviewed.A total of 91 patients with bacteremia due to Aeromonas species were identified. In addition to 16 (17.6% primary bacteremia, the most common source of secondary infection is peritonitis (n = 27, 29.7%, followed by biliary tract infection (n = 18, 19.8%, and SSTI (n = 12, 13.2%, pneumonia (n = 9, 9.9%, catheter-related bloodstream infection (n =  5, 5.5%, and genitourinary tract infection (n = 4, 4.4%. A. hydrophila (n = 35, 38.5% was the most common pathogen, followed by A. veronii biovar sobria (n = 31, 34.1%, A. caviae (n = 14, 15.4%, and A. veronii biovar veronii (n = 9, 9.9%. Forty-three (47.3% patients were classified as healthcare-associated infections (HCAI causes by Aeromonas species, and patients with HCAI were more likely to have cancer, and receive immunosuppressant than patients with community-acquired bacteremia. The overall outcomes, including rate of ICU admission, acute respiratory failure, and mortality were 33.3%, 28.6%, and 23.1%, respectively. Multivariate analysis showed that the in-hospital day mortality was significantly associated only with underlying cancer (P <.001, and initial shock (P <.001.Aeromonas species should be considered one of the causative pathogens of healthcare-associated bacteremia, especially in immunocompromised patients. In addition, it can be associated with high fatality. Cancer and initial shock were the poor prognostic factors.

  9. Genetic Diversity and Symbiotic Efficiency of Indigenous Common Bean Rhizobia in Croatia

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    Ines Pohajda

    2016-01-01

    Full Text Available Nodule bacteria (rhizobia in symbiotic associations with legumes enable considerable entries of biologically fixed nitrogen into soil. Efforts are therefore made to intensify the natural process of symbiotic nitrogen fixation by legume inoculation. Studies of field populationsof rhizobia open up the possibility to preserve and probably exploit some indigenous strains with hidden symbiotic or ecological potentials. The main aim of the present study is to determine genetic diversity of common bean rhizobia isolated from different field sites in central Croatia and to evaluate their symbiotic efficiency and compatibility with host plants. The isolation procedure revealed that most soil samples contained no indigenous common bean rhizobia. The results indicate that the cropping history had a significant impact on the presence of indigenous strains. Although all isolates were found to belong to species Rhizobium leguminosarum, significant genetic diversity at the strain level was determined. Application of both random amplifi cation of polymorphic DNA (RAPD and enterobacterial repetitive intergenic consensus–polymerase chain reaction (ERIC-PCR methods resulted in similar grouping of strains. Symbiotic efficiency of indigenous rhizobia as well as their compatibility with two commonly grown bean varieties were tested in field experiments. Application of indigenous rhizobial strains as inoculants resulted in significantly different values of nodulation, seed yield as well as plant nitrogen and seed protein contents. The most abundant nodulation and the highest plant nitrogen and protein contents were determined in plants inoculated with R. leguminosarum strains S17/2 and S21/6. Although, in general, the inoculation had a positive impact on seed yield, differences depending on the applied strain were not determined. The overall results show the high degree of symbiotic efficiency of the specific indigenous strain S21/6. These results indicate different

  10. Products of Dark CO2 Fixation in Pea Root Nodules Support Bacteroid Metabolism 1

    Science.gov (United States)

    Rosendahl, Lis; Vance, Carroll P.; Pedersen, Walther B.

    1990-01-01

    Products of the nodule cytosol in vivo dark [14C]CO2 fixation were detected in the plant cytosol as well as in the bacteroids of pea (Pisum sativum L. cv “Bodil”) nodules. The distribution of the metabolites of the dark CO2 fixation products was compared in effective (fix+) nodules infected by a wild-type Rhizobium leguminosarum (MNF 300), and ineffective (fix−) nodules of the R. leguminosarum mutant MNF 3080. The latter has a defect in the dicarboxylic acid transport system of the bacterial membrane. The 14C incorporation from [14C]CO2 was about threefold greater in the wild-type nodules than in the mutant nodules. Similarly, in wild-type nodules the in vitro phosphoenolpyruvate carboxylase activity was substantially greater than that of the mutant. Almost 90% of the 14C label in the cytosol was found in organic acids in both symbioses. Malate comprised about half of the total cytosol organic acid content on a molar basis, and more than 70% of the cytosol radioactivity in the organic acid fraction was detected in malate in both symbioses. Most of the remaining 14C was contained in the amino acid fraction of the cytosol in both symbioses. More than 70% of the 14C label found in the amino acids of the cytosol was incorporated in aspartate, which on a molar basis comprised only about 1% of the total amino acid pool in the cytosol. The extensive 14C labeling of malate and aspartate from nodule dark [14C]CO2 fixation is consistent with the role of phosphoenolpyruvate carboxlase in nodule dark CO2 fixation. Bacteroids from the effective wild-type symbiosis accumulated sevenfold more 14C than did the dicarboxylic acid transport defective bacteroids. The bacteroids of the effective MNF 300 symbiosis contained the largest proportion of the incorporated 14C in the organic acids, whereas ineffective MNF 3080 bacteroids mainly contained 14C in the amino acid fraction. In both symbioses a larger proportion of the bacteroid 14C label was detected in malate and aspartate

  11. Genetic Diversity and Symbiotic Efficiency of Indigenous Common Bean Rhizobia in Croatia.

    Science.gov (United States)

    Pohajda, Ines; Babić, Katarina Huić; Rajnović, Ivana; Kajić, Sanja; Sikora, Sanja

    2016-12-01

    Nodule bacteria (rhizobia) in symbiotic associations with legumes enable considerable entries of biologically fixed nitrogen into soil. Efforts are therefore made to intensify the natural process of symbiotic nitrogen fixation by legume inoculation. Studies of field populations of rhizobia open up the possibility to preserve and probably exploit some indigenous strains with hidden symbiotic or ecological potentials. The main aim of the present study is to determine genetic diversity of common bean rhizobia isolated from different field sites in central Croatia and to evaluate their symbiotic efficiency and compatibility with host plants. The isolation procedure revealed that most soil samples contained no indigenous common bean rhizobia. The results indicate that the cropping history had a significant impact on the presence of indigenous strains. Although all isolates were found to belong to species Rhizobium leguminosarum , significant genetic diversity at the strain level was determined. Application of both random amplification of polymorphic DNA (RAPD) and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC- -PCR) methods resulted in similar grouping of strains. Symbiotic efficiency of indigenous rhizobia as well as their compatibility with two commonly grown bean varieties were tested in field experiments. Application of indigenous rhizobial strains as inoculants resulted in significantly different values of nodulation, seed yield as well as plant nitrogen and seed protein contents. The most abundant nodulation and the highest plant nitrogen and protein contents were determined in plants inoculated with R. leguminosarum strains S 17/2 and S 21/6 . Although, in general, the inoculation had a positive impact on seed yield, differences depending on the applied strain were not determined. The overall results show the high degree of symbiotic efficiency of the specific indigenous strain S 21/6 . These results indicate different symbiotic

  12. Chemical characterization of bioactive compounds from the endophytic fungus Diaporthe helianthi isolated from Luehea divaricata

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    Vânia Specian

    2012-09-01

    Full Text Available Endophytic microorganisms, defined as fungi or bacteria that colonize the interior of plants without causing any immediate negative effects or damages, have reciprocal relationships with host plants. In some cases their presence is beneficial to the host due to the synthesis of bioactive compounds, among which several alcohols, esters, ketones and others that may react with other compounds and may be lethal to pathogenic microorganisms. Diaporthe helianthi (Phomopsis helianthi in its anamorphic phase is available worldwide, especially in Europe, Asia and America. Isolated in Europe as an agent of the sunflower stem cancer, it has also been endophytically isolated from tropical and temperate plants. A D. helianthi strain isolated from Luehea divaricata has been employed in current research. An investigation of the secondary metabolite from D. helianthi by CC and NMR of ¹H and 13C yielded the separation of 10 fractions and the identification of the phenolic compound 2(-4 hydroxyphenyl-ethanol (Tyrosol. Its antimicrobial reaction was tested and the ensuing antagonistic effects on the human pathogenic bacteria Enterococcus hirae, Escherichia coli, Micrococcus luteus, Salmonella typhi, Staphylococcus aureus, phytopathogenic Xanthomonas asc. phaseoli and phytopathogenic fungi were demonstrated. Results show that bioactive compounds and Tyrosol produced by D. helianthi have a biotechnological potential.

  13. Defense reactions of bean genotypes to bacterial pathogens in controlled conditions

    Science.gov (United States)

    Uysal, B.; Bastas, K. K.

    2018-03-01

    This study was focused on the role of antioxidant enzymes and total protein in imparting resistance against common bacterial blight caused by Xanthomonas axonopodis pv. phaseoli (Xap) and halo blight caused by Pseudomonas syringae pv. phaseolicola (Psp) in bean. Activities of Ascorbate peroxidase (APX), Catalase (CAT) and total protein were studied in resistant and susceptible bean genotypes. Five-day-old seedlings were inoculated with a bacterial suspension (108 CFU ml-1) and harvested at different time intervals (0, 12, 24 and 36 up to 72 h) under controlled growing conditions and assayed for antioxidant enzymes and total protein. Temporal increase of CAT, APX enzymes activities showed maximum activity at 12 h after both pathogens inoculation (hpi) in resistant cultivar, whereas in susceptible it increased at 72 h after both pathogens inoculation for CAT and 12, 24 h for APX enzymes. Maximum total protein activities were observed at 12 h and 24 h respectively after Xap, Psp inoculation (hpi) in resistant and maximum activities were observed at 24 h and 72 h respectively after Xap, Psp inoculation (hpi) in susceptible. Increase of antioxidant enzyme and total protein activities might be an important component in the defense strategy of resistance and susceptible bean genotypes against the bacterial infection. These findings suggest that disease protection is proportional to the amount of enhanced CAT, APX enzyme and total protein activity.

  14. Induced mutations for disease resistance and other agronomic characteristics in bean (Phaseolus vulgaris L.) and soybean (Glycine max (L.) Merrill)

    International Nuclear Information System (INIS)

    Tulmann Neto, A.; Menten, J.O.M.; Ando, A.; Alberini, J.; Peixoto, T.C.

    1988-01-01

    The present research project aims to induce mutations with resistance to Xanthomonas phaseoli (common blight) and golden mosaic virus (GMV) in bean and resistance to Phakopsora pachyrhizi (rust) and Brazilian bud blight in soybean. At the same time, other mutant types of interest were selected. Gamma rays and ethyl methane-sulphonate (EMS) were generally utilized as mutagenic agents and seeds of several cultivars from both crops were treated. The selection was made at the M 2 or M 3 generation, utilizing progeny or mixtures of seeds from bulk. Screening was carried out in the field, greenhouse or insectary (according to the disease). Priority was given to GMV in bean and about 235,850 plants were observed in the field and 67,500 in the insectary. Only one plant showing mild GMV symptoms was obtained. However, owing to negative pleiotropic effects, this mutant could not be used. Concerning the other diseases, there are some selected plants that still require better observation before reporting that progress has been made. With regard to other mutant types, earliness was obtained in soybean and a bush variety and an earlier mutant was selected in bean. This mutant has already been utilized by breeders in cross-breeding and is being multiplied to be experimentally utilized by farmers under special conditions of cultivation. In soybean, preliminary yield trials are under way, and include some of the early mutants obtained. (author). 26 refs, 20 tabs

  15. Non-host disease resistance response in pea (Pisum sativum) pods: Biochemical function of DRR206 and phytoalexin pathway localization.

    Science.gov (United States)

    Seneviratne, Herana Kamal; Dalisay, Doralyn S; Kim, Kye-Won; Moinuddin, Syed G A; Yang, Hong; Hartshorn, Christopher M; Davin, Laurence B; Lewis, Norman G

    2015-05-01

    Continually exposed to potential pathogens, vascular plants have evolved intricate defense mechanisms to recognize encroaching threats and defend themselves. They do so by inducing a set of defense responses that can help defeat and/or limit effects of invading pathogens, of which the non-host disease resistance response is the most common. In this regard, pea (Pisum sativum) pod tissue, when exposed to Fusarium solani f. sp. phaseoli spores, undergoes an inducible transcriptional activation of pathogenesis-related genes, and also produces (+)-pisatin, its major phytoalexin. One of the inducible pathogenesis-related genes is Disease Resistance Response-206 (DRR206), whose role in vivo was unknown. DRR206 is, however, related to the dirigent protein (DP) family. In this study, its biochemical function was investigated in planta, with the metabolite associated with its gene induction being pinoresinol monoglucoside. Interestingly, both pinoresinol monoglucoside and (+)-pisatin were co-localized in pea pod endocarp epidermal cells, as demonstrated using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging. In addition, endocarp epidermal cells are also the site for both chalcone synthase and DRR206 gene expression. Taken together, these data indicate that both (+)-pisatin and pinoresinol monoglucoside function in the overall phytoalexin responses. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Anatomy of a nonhost disease resistance response of pea to Fusarium solani: PR gene elicitation via DNase, chitosan and chromatin alterations.

    Directory of Open Access Journals (Sweden)

    Lee A Hadwiger

    2015-06-01

    Full Text Available Of the multiplicity of plant pathogens in nature, only a few are virulent on a given plant species. Conversely, plants develop a rapid nonhost resistance response to the majority of the pathogens. The anatomy of the nonhost resistance of pea endocarp tissue against a pathogen of bean, Fusarium solani f.sp. phaseoli (Fsph and the susceptibility of pea to F. solani f sp. pisi (Fspi has been described cytologically, biochemically and molecular-biologically. Cytological changes have been followed by electron microscope and stain differentiation under white and UV light. The induction of changes in transcription, protein synthesis, expression of pathogenesis-related (PR genes, and increases in metabolic pathways culminating in low molecular weight, antifungal compounds are described biochemically. Molecular changes initiated by fungal signals to host organelles, primarily to the chromatin within host nuclei, are identified according to source of the signal and the mechanisms utilized in activating defense genes. The functions of some PR genes are defined. A hypothesis based on this data is developed to explain both why fungal growth is suppressed in nonhost resistance and why growth can continue in a susceptible reaction.

  17. Anatomy of a nonhost disease resistance response of pea to Fusarium solani: PR gene elicitation via DNase, chitosan and chromatin alterations

    Science.gov (United States)

    Hadwiger, Lee A.

    2015-01-01

    Of the multiplicity of plant pathogens in nature, only a few are virulent on a given plant species. Conversely, plants develop a rapid “nonhost” resistance response to the majority of the pathogens. The anatomy of the nonhost resistance of pea endocarp tissue against a pathogen of bean, Fusarium solani f.sp. phaseoli (Fsph) and the susceptibility of pea to F. solani f sp. pisi (Fspi) has been described cytologically, biochemically and molecular-biologically. Cytological changes have been followed by electron microscope and stain differentiation under white and UV light. The induction of changes in transcription, protein synthesis, expression of pathogenesis-related (PR) genes, and increases in metabolic pathways culminating in low molecular weight, antifungal compounds are described biochemically. Molecular changes initiated by fungal signals to host organelles, primarily to chromatin within host nuclei, are identified according to source of the signal and the mechanisms utilized in activating defense genes. The functions of some PR genes are defined. A hypothesis based on this data is developed to explain both why fungal growth is suppressed in nonhost resistance and why growth can continue in a susceptible reaction. PMID:26124762

  18. Effect of participatory selection of varieties on the identification of outstanding common bean genotypes (Phaseolus vulgaris L.

    Directory of Open Access Journals (Sweden)

    Alexis Lamz Piedra

    2017-10-01

    Full Text Available Among the main factors affecting bean production is poor distribution of varieties for different environmental conditions in which its are grown. The aim of this work was to evaluate the impact of participatory selection of foreign genetic materials and national commercial and pre-commercial common bean in identifying genotypes for their outstanding performance and resistance to common bacteriosis (Xanthomonas campestris pv. phaseoli (Smith Dye (Xcp. In the "El Mulato" farm belonging to the Empowered Cooperative of Credit and Services (CCSF "Orlando Cuellar" in the municipality San José de las Lajas, Mayabeque, two experiments were conducted. In the first one, 15 genotypes were planted in 13 September 2014 (early season in experimental plots to develop a diversity Fair and evaluate the natural incidence of common bean bacteriosis. In the second experiment, they were sown on 25 December (late season the materials selected by farmers with superior agronomic performance (7 genotypes to validate the stability of its performance. Among the results, an effective range of 93,33 % between the selected materials and selective criteria that this diversity was identified were high performance, resistance to common bacteriosis and color of beans. It was found that the selection of the diversity of beans by farmers is not influenced by the origin of materials and participatory selection identified common bean genotypes with high yield potential and stability between planting seasons.

  19. SOJA IAC/IAS-5: CULTIVAR RESISTENTE AO CANCRO DA HASTE

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    MASCARENHAS HIPÓLITO ASSUNÇÃO ANTONIO

    1998-01-01

    Full Text Available Há aproximadamente oito anos, o cancro da haste (Phomopsis phaseoli f. sp. meridionalis/Diaporthe phaseolorum f. sp. meridionalis da soja tem sido o principal fator limitante da produtividade em áreas de manifestação da doença. O seu controle mediante cultivares resistentes é o método mais econômico. Num ensaio em casa de vegetação, visando avaliar o efeito de doses de potássio no controle do cancro da haste, no cultivar IAS-5, moderadamente resistente, verificou-se haver plantas altamente resistentes, comprovadas em três gerações, todas inoculadas com o fungo causador da doença. A importância da obtenção do IAS-5 e de sua multiplicação deve-se a ser o mais cultivado no Estado de São Paulo e apresentar resistência ao cancro da haste.

  20. Association of Effector Six6 with Vascular Wilt Symptoms Caused by Fusarium oxysporum on Soybean.

    Science.gov (United States)

    Lanubile, Alessandra; Ellis, Margaret L; Marocco, Adriano; Munkvold, Gary P

    2016-11-01

    The Fusarium oxysporum species complex (FOSC) is a widely distributed group of fungi that includes both pathogenic and nonpathogenic isolates. In a previous study, isolates within the FOSC collected primarily from soybean were assessed for the presence of 12 fungal effector genes. Although none of the assayed genes was significantly associated with wilt symptoms on soybean, the secreted in xylem 6 (Six6) gene was present only in three isolates, which all produced high levels of vascular wilt on soybean. In the current study, a collection of F. oxysporum isolates from soybean roots and F. oxysporum f. sp. phaseoli isolates from common bean was screened for the presence of the Six6 gene. Interestingly, all isolates for which the Six6 amplicon was generated caused wilt symptoms on soybean, and two-thirds of the isolates showed high levels of aggressiveness, indicating a positive association between the presence of the effector gene Six6 and induction of wilt symptoms. The expression profile of the Six6 gene analyzed by quantitative reverse-transcription polymerase chain reaction revealed an enhanced expression for the isolates that caused more severe wilt symptoms on soybean, as established by the greenhouse assay. These findings suggest the suitability of the Six6 gene as a possible locus for pathogenicity-based molecular diagnostics across the various formae speciales.

  1. Recombinant expression, purification and biochemical characterization of kievitone hydratase from Nectria haematococca.

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    Matthias Engleder

    Full Text Available Kievitone hydratase catalyzes the addition of water to the double bond of the prenyl moiety of plant isoflavonoid kievitone and, thereby, forms the tertiary alcohol hydroxy-kievitone. In nature, this conversion is associated with a defense mechanism of fungal pathogens against phytoalexins generated by host plants after infection. As of today, a gene sequence coding for kievitone hydratase activity has only been identified and characterized in Fusarium solani f. sp. phaseoli. Here, we report on the identification of a putative kievitone hydratase sequence in Nectria haematococca (NhKHS, the teleomorph state of F. solani, based on in silico sequence analyses. After heterologous expression of the enzyme in the methylotrophic yeast Pichia pastoris, we have confirmed its kievitone hydration activity and have assessed its biochemical properties and substrate specificity. Purified recombinant NhKHS is obviously a homodimeric glycoprotein. Due to its good activity for the readily available chalcone derivative xanthohumol (XN, this compound was selected as a model substrate for biochemical studies. The optimal pH and temperature for hydratase activity were 6.0 and 35°C, respectively, and apparent Vmax and Km values for hydration of XN were 7.16 μmol min-1 mg-1 and 0.98 ± 0.13 mM, respectively. Due to its catalytic properties and apparent substrate promiscuity, NhKHS is a promising enzyme for the biocatalytic production of tertiary alcohols.

  2. Hairy root transgene expression analysis of a secretory peroxidase (PvPOX1) from common bean infected by Fusarium wilt.

    Science.gov (United States)

    Xue, Renfeng; Wu, Xingbo; Wang, Yingjie; Zhuang, Yan; Chen, Jian; Wu, Jing; Ge, Weide; Wang, Lanfen; Wang, Shumin; Blair, Matthew W

    2017-07-01

    Plant peroxidases (POXs) are one of the most important redox enzymes in the defense responses. However, the large number of different plant POX genes makes it necessary to carefully confirm the function of each paralogous POX gene in specific tissues and disease interactions. Fusarium wilt is a devastating disease of common bean caused by Fusarium oxysporum f. sp. phaseoli. In this study, we evaluated a peroxidase gene, PvPOX1, from a resistant common bean genotype, CAAS260205 and provided direct evidence for PvPOX1's role in resistance by transforming the resistant allele into a susceptible common bean genotype, BRB130, via hairy root transformation using Agrobacterium rhizogenes. Analysis of PvPOX1 gene over-expressing hairy roots showed it increased resistance to Fusarium wilt both in the roots and the rest of transgenic plants. Meanwhile, the PvPOX1 expressive level, the peroxidase activity and hydrogen peroxide (H 2 O 2 ) accumulation were also enhanced in the interaction. The result showed that the PvPOX1 gene played an essential role in Fusarium wilt resistance through the occurrence of reactive oxygen species (ROS) induced hypersensitive response. Therefore, PvPOX1 expression was proven to be a valuable gene for further analysis which can strengthen host defense response against Fusarium wilt through a ROS activated resistance mechanism. Copyright © 2017 Elsevier B.V. All rights reserved.

  3. Common bean genotypes for agronomic and market-related traits in VCU trials

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    Alisson Fernando Chiorato

    2015-02-01

    Full Text Available Value for Cultivation and Use (VCU trials are undertaken when evaluating improved common bean (Phaseolus vulgaris L. lines, and knowledge of agronomic and market-related traits and disease reaction is instrumental in making cultivar recommendations. This study evaluates the yield, cooking time, grain color and reaction to anthracnose (Colletotrichum lindemuthianum, Fusarium wilt (Fusarium oxysporum f. sp. phaseoli and Curtobacterium wilt (Curtobacterium flaccumfaciens pv. flaccumfaciens of 25 common bean genotypes derived from the main common bean breeding programs in Brazil. Seventeen VCU trials were carried out in the rainy season, dry season and winter season from 2009 to 2011 in the state of São Paulo. Analyses of grain color and cooking time were initiated 60 days after harvest, and disease reaction analyses were performed in the laboratory under controlled conditions. In terms of yield, no genotype superior to the controls was observed for any of the seasons under consideration. Grains from the dry season exhibited better color, while the rainy season led to the shortest cooking times. The following genotypes BRS Esteio, BRS Esplendor and IAC Imperador were resistant to anthracnose, Fusarium wilt and Curtobacterium wilt and, in general, genotypes with lighter-colored grains were more susceptible to anthracnose and Fusarium wilt.

  4. Beginning to understand the role of sugar carriers in Colletotrichum lindemuthianum: the function of the gene mfs1.

    Science.gov (United States)

    Pereira, Monalessa Fábia; de Araújo Dos Santos, Carolina Maria; de Araújo, Elza Fernandes; de Queiroz, Marisa Vieira; Bazzolli, Denise Mara Soares

    2013-02-01

    Fungi of the Colletotrichum genus are among the most prominent phytopathogens that cause diseases with a considerable economic impact, such as anthracnose. The hemibiotrophic fungus Colletotrichum lindemuthianum (teleomorph Glomerella cingulata f. sp. phaseoli) is the causal agent of the anthracnose of the common bean; and similarly to other phytopathogens, it uses multiple strategies to gain access to different carbon sources from its host. In this study, we examine mfs1, a newly identified C. lindemuthianum hexose transporter. The mfs1 gene is expressed only during the necrotrophic phase of the fungus' interaction within the plant and allows it to utilize the available sugars during this phase. The deletion of mfs1 gene resulted in differential growth of the fungus in a medium that contained glucose, mannose or fructose as the only carbon source. This study is the first to describe a hexose transporter in the hemibiotrophic pathogen C. lindemuthianum and to demonstrate the central role of this protein in capturing carbon sources during the necrotrophic development of the plant/pathogen interaction.

  5. Anatomy of a nonhost disease resistance response of pea to Fusarium solani: PR gene elicitation via DNase, chitosan and chromatin alterations.

    Science.gov (United States)

    Hadwiger, Lee A

    2015-01-01

    Of the multiplicity of plant pathogens in nature, only a few are virulent on a given plant species. Conversely, plants develop a rapid "nonhost" resistance response to the majority of the pathogens. The anatomy of the nonhost resistance of pea endocarp tissue against a pathogen of bean, Fusarium solani f.sp. phaseoli (Fsph) and the susceptibility of pea to F. solani f sp. pisi (Fspi) has been described cytologically, biochemically and molecular-biologically. Cytological changes have been followed by electron microscope and stain differentiation under white and UV light. The induction of changes in transcription, protein synthesis, expression of pathogenesis-related (PR) genes, and increases in metabolic pathways culminating in low molecular weight, antifungal compounds are described biochemically. Molecular changes initiated by fungal signals to host organelles, primarily to chromatin within host nuclei, are identified according to source of the signal and the mechanisms utilized in activating defense genes. The functions of some PR genes are defined. A hypothesis based on this data is developed to explain both why fungal growth is suppressed in nonhost resistance and why growth can continue in a susceptible reaction.

  6. Potentiel infectieux des sols de cultures de tomate (Lycopersicon esculentum Mill. sous abri et incidence de l'âge de repiquage sur la vigueur des plants vis-à-vis de Pythium sp. à Songon-Dabou en Côte d'Ivoire

    Directory of Open Access Journals (Sweden)

    Koné, D.

    2008-01-01

    Full Text Available Potentiel Infectious of Soils Fields of Tomato (Lycopersicon esculentum Mill. in Greenhouse and the Age Incidence Facing Pythium sp. on the Plants at Songon-Dabou in Ivory Coast. Songon-Dabou area represents one of the spheres activities of tomato in Ivory Coast. The production of tomato is strongly influenced by some of viruses, bacteria and soil born fungi. A study was conducted on the soils of different fields with three varieties of tomato (Caraïbo, Mongal and Tropimech to identify the most susceptible variety to soil born fungi at Songon-Dabou. By baiting with 20 days old seedlings, 4 genus fungi of Pythium, Fusarium, Sclerotium and Macrophomina have been isolated. Pythium sp. presented the greatest attack (32% while Macrophomina phaseoli was isolated for the first time on market gardening in Ivory Coast. Tomato plants have been inoculated by Pythium at different stages obtained from seedling to 25 days old nursery seedlings. That experiment was repeated three times. This work was conducted in greenhouse during 26 days after transplantation of the plants in the pots. These results confirm that Pythium sp. is a fungus of damping off (100% and that age of transplantation reduces the rate of mortality (less than 20%.

  7. Chemical Composition and Antibacterial Activity of Essential Oils of Tagetes minuta (Asteraceae) against Selected Plant Pathogenic Bacteria

    Science.gov (United States)

    Wagacha, John M.; Dossaji, Saifuddin F.

    2016-01-01

    The objective of this study was to determine the chemical composition and antibacterial activity of essential oils (EOs) of Tagetes minuta against three phytopathogenic bacteria Pseudomonas savastanoi pv. phaseolicola, Xanthomonas axonopodis pv. phaseoli, and Xanthomonas axonopodis pv. manihotis. The essential oils were extracted using steam distillation method in a modified Clevenger-type apparatus while antibacterial activity of the EOs was evaluated by disc diffusion method. Gas chromatography coupled to mass spectrometry (GC/MS) was used for analysis of the chemical profile of the EOs. Twenty compounds corresponding to 96% of the total essential oils were identified with 70% and 30% of the identified components being monoterpenes and sesquiterpenes, respectively. The essential oils of T. minuta revealed promising antibacterial activities against the test pathogens with Pseudomonas savastanoi pv. phaseolicola being the most susceptible with mean inhibition zone diameters of 41.83 and 44.83 mm after 24 and 48 hours, respectively. The minimum inhibitory concentrations and minimum bactericidal concentrations of the EOs on the test bacteria were in the ranges of 24–48 mg/mL and 95–190 mg/mL, respectively. These findings provide a scientific basis for the use of T. minuta essential oils as a botanical pesticide for management of phytopathogenic bacteria. PMID:27721831

  8. Cloning and characterization of a galactitol 2-dehydrogenase from Rhizobium legumenosarum and its application in D-tagatose production.

    Science.gov (United States)

    Jagtap, Sujit Sadashiv; Singh, Ranjitha; Kang, Yun Chan; Zhao, Huimin; Lee, Jung-Kul

    2014-05-10

    Galactitol 2-dehydrogenase (GDH) belongs to the protein subfamily of short-chain dehydrogenases/reductases and can be used to produce optically pure building blocks and for the bioconversion of bioactive compounds. An NAD(+)-dependent GDH from Rhizobium leguminosarum bv. viciae 3841 (RlGDH) was cloned and overexpressed in Escherichia coli. The RlGDH protein was purified as an active soluble form using His-tag affinity chromatography. The molecular mass of the purified enzyme was estimated to be 28kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and 114kDa by gel filtration chromatography, suggesting that the enzyme is a homotetramer. The enzyme has an optimal pH and temperature of 9.5 and 35°C, respectively. The purified recombinant RlGDH catalyzed the oxidation of a wide range of substrates, including polyvalent aliphatic alcohols and polyols, to the corresponding ketones and ketoses. Among various polyols, galactitol was the preferred substrate of RlGDH with a Km of 8.8mM, kcat of 835min(-1) and a kcat/Km of 94.9min(-1)mM(-1). Although GDHs have been characterized from a few other sources, RlGDH is distinguished from other GDHs by its higher specific activity for galactitol and broad substrate spectrum, making RlGDH a good choice for practical applications. Copyright © 2014 Elsevier Inc. All rights reserved.

  9. Biologic fixation of nitrogen in irradiated rhizobium strips; Fixacao biologica do nitrogenio em estirpes de rizobianas irradiadas

    Energy Technology Data Exchange (ETDEWEB)

    Caribe, Rebeka Alves; Colaco, Waldeciro [Pernambuco Univ., Recife, PE (Brazil). Dept. de Energia Nuclear

    2002-07-01

    Native Rhizobium sp. and Bradyrhizobium sp. isolates from the root nodules of bean and cowpea were selected. Six isolates, and the SEMIA 4077 (R. leguminosarum bv. phaseolus) and SEMIA 6145 (Bradyrhizobium sp) strains used as references, were irradiated with ultraviolet light (R-uv) and gamma rays (R-{gamma}). The D{sub 37} values for the rhizobial strain SEMIA 4077 were 43 J.m{sup -2} (UV) and 32 Gy (R-{gamma}) and for the SEMIA 6145 were 45 J.m{sup -2} (UV) and 35 Gy (R-gamma). Through a greenhouse experiment the irradiated isolates were inoculated on bean (P. vulgaris L., cv. Princesa) and on cowpea [Vigna unguiculata, (L.) Walp, cv. IPA-206] seedlings, in an attempt to evaluate the sensitivity of the host plants, and possible effects on their nodulation. Differences in responses to nodulation due to the effect of irradiation were observed for the isolates tested. Significantly differences were observed only for nodules dry matter yield of the IPA-206 cultivar. Gamma irradiated treatment were statistically superior to treatments with ultraviolet light in relation. (author)

  10. Estimation of the effect of radionuclide contamination on Vicia sativa L. induction of chlorophyll fluorescence parameters using "Floratest" optical biosensor

    Science.gov (United States)

    Ruban, Yu.; Illienko, V.; Nesterova, N.; Pareniuk, O.; Shavanova, K.

    2017-12-01

    The presented research was aimed to determine the parameters of chlorophyll fluorescence (IChH) curve induction for Vicia sativa L. that were grown on radionuclide contaminated soils by using "Floratest" fluorometer. Plants were inoculated with 5 species of bacteria that might potentially block radionuclide uptake (Agrobacterium radiobacter IMBB-7246, Azotobacter chroococcum UKMB-6082, A. chroococcum UKMB-6003, Bacillus megaterium UKMB-5724, Rhizobium leguminosarum bv. viceae) and grown in sod-podzolic, chernozem and peat-bog soils, contaminated with 137Cs (4000±340 Bq/kg). As a result of research, it was determined that the most stressful factors for vetch plants are combination of soil radionuclide and presence of Bacillus megaterium UKM B-5724, as the number of inactive chlorophyll increased. In addition, the vetch plants significantly increased fixed level of fluorescence (Fst) under the influence of radioactive contamination in presence of Bacillus megaterium UKM B-5724, indicating inhibition of photosynthetic reactions. Other bacteria showed radioprotective properties in almost all types of soil.

  11. A novel type of DNA-binding protein interacts with a conserved sequence in an early nodulin ENOD12 promoter

    DEFF Research Database (Denmark)

    Christiansen, H; Hansen, A C; Vijn, I

    1996-01-01

    The pea genes PsENOD12A and PsENOD12B are expressed in the root hairs shortly after infection with the nitrogen-fixing bacterium Rhizobium leguminosarum bv. viciae or after application of purified Nod factors. A 199 bp promoter fragment of the PsENOD12B gene contains sufficient information for Nod...... factor-induced tissue-specific expression. We have isolated a Vicia sativa cDNA encoding a 1641 amino acid protein, ENBP1, that interacts with the 199 bp ENOD12 promoter. Two different DNA-binding domains were identified in ENBP1. A domain containing six AT-hooks interacts specifically with an AT...... of the ENBP1 transcript in cells expressing ENOD12 strongly suggest that ENBP1 is a transcription factor involved in the regulation of ENOD12. Finally, the C-terminal region of ENBP1 shows strong homology to a protein from rat that is specifically expressed in testis tissue. Udgivelsesdato: 1996-Dec...

  12. Draft Genome Sequence of the Nitrogen-Fixing Rhizobium sullae Type Strain IS123T Focusing on the Key Genes for Symbiosis with its Host Hedysarum coronarium L.

    Directory of Open Access Journals (Sweden)

    Gaurav Sablok

    2017-07-01

    Full Text Available The prominent feature of rhizobia is their molecular dialogue with plant hosts. Such interaction is enabled by the presence of a series of symbiotic genes encoding for the synthesis and export of signals triggering organogenetic and physiological responses in the plant. The genome of the Rhizobium sullae type strain IS123T nodulating the legume Hedysarum coronarium, was sequenced and resulted in 317 scaffolds for a total assembled size of 7,889,576 bp. Its features were compared with those of genomes from rhizobia representing an increasing gradient of taxonomical distance, from a conspecific isolate (Rhizobium sullae WSM1592, to two congeneric cases (Rhizobium leguminosarum bv. viciae and Rhizobium etli and up to different genera within the legume-nodulating taxa. The host plant is of agricultural importance, but, unlike the majority of other domesticated plant species, it is able to survive quite well in the wild. Data showed that that the type strain of R. sullae, isolated from a wild host specimen, is endowed with a richer array of symbiotic genes in comparison to other strains, species or genera of rhizobia that were rescued from domesticated plant ecotypes. The analysis revealed that the bacterium by itself is incapable of surviving in the extreme conditions that its host plant can tolerate. When exposed to drought or alkaline condition, the bacterium depends on its host to survive. Data are consistent with the view of the plant phenotype as the primary factor enabling symbiotic nitrogen fixing bacteria to survive in otherwise limiting environments.

  13. Effect of exogenous supply of boron on nodule development in pea (pisum sativum L.)

    International Nuclear Information System (INIS)

    Qasim, M.; Khan, Z.U.D.; Syed, H.R.; Mehmood, F.

    2011-01-01

    Exogenous supply of boron was evaluated on the seeds of Pisum sativum L. inoculated with Rhizobium leguminosarum and grown in aqua-culture. The liquid nutrition medium contained six boron concentrations (control, 1, 1.86, 2.86, 3.86, and 4.86 mg L/sup -1/ ). The results obtained from the present work showed that number of nodules, size of nodules and weight of boron treatment. Toxic effect of high concentration of boron (4.86 mg L/sup -1/ ) nodules exhibit great improvement in 2.86 mg L /sup-1/ was also recorded. There was significant reduction as compare to control in nodule weight, size and numbers. This fact becomes clear while observing the nodules growing in the boron free culture, which did not develop extensively. The comparison of the transverse sections of root-nodule area, from nutrient solutions with various boron concentrations showed that in the absence of boron, there was a considerable hypertrophy in cambial cells and a frequent disintegration of phloem and ground tissue along with xylem. The disintegration of the tissue seems to be linked with nodule bacteria. The nodular bacteria are considered to assume a parasitic habit in the absence of boron. (author)

  14. VARIATION IN RHIZOBIUM GROWTH DUE TO SEED AND ROOT EXUDATES RELEASED FROM GAMMA IRRADIATED GLYCINE MAX SEEDS

    International Nuclear Information System (INIS)

    KAMEL, H.A.; ASKER, M.M.S

    2008-01-01

    In this study, seeds of Glycine max Giza 122 were irradiated with gamma rays from 60 Co source at various doses (10 to 200 Gy), sterilized and soaked into an aerated solution of CaSO 4 (1 mmol and pH 6.5). The capacities of the released seed exudates (SEs) and root exudates (REs) to promote Rhizobium leguminosarum growth were investigated as well as biochemical analysis of the exudates was carried out. SE of both control and gamma irradiated seeds resulted in a higher Rhizobium population and polysaccharide production than RE. Relative to control, the highly effective doses in Rhizobium growth and polysaccharide production were 25 and 200 Gy; the former was a promoter while the later was an inhibitor. HPLC analysis of soluble carbohydrates revealed the presence of glucose (Glu), rhamnose (Rha) and fructose (Fru) in the SE and RE. Protein content in SE was lower than that in RE; the highest values were due to 10 Gy and 25 Gy in SE and RE, respectively. Free amino acids content in SE was increased up to 25 Gy then decreased while RE was increased by increasing gamma doses from 10 to 200 Gy

  15. Effect of inoculation and nitrogen top-dressing on yields and fodder value of winter pea cv. Maksimirski ozimi in triticale cv. Clercal mixture

    Directory of Open Access Journals (Sweden)

    Darko Uher

    2006-07-01

    Full Text Available Two year field trials (1999-2001 were aimed to determine the inoculation seed winter pea and nitrogen top-dressing effect on number and active nodules of pea root and also on the green mass and dry matter yield of winter pea cv. Maksimirski ozimi and triticale cv. Clercal mixture. Immediately before sowing the inoculation of pea seeds was done by the indigenous variety of Rhizobium leguminosarum bv. viciae which belongs to the collection of Department of Microbiology at the Faculty of Agriculture University of Zagreb. The highest total nodule number on pea root (134 was determined on the inoculated variant as well as active nodule (120. Average mixture green mass yields were ranging from 20.9 t ha-1 (control up to 30.1 t ha-1 (inoculation + nitrogen top-dressing. Total dry matter yields were ranging from 3.71 t ha-1 (control up to 5.36 t ha-1 (nitrogen top-dressing. Pea crude protein yields in 2001 were ranging from 500 kg ha-1 (nitrogen top-dressing up to 642 kg ha-1 (inoculation and for triticale, those values were from 322 kg ha-1 (control up to 498 kg ha-1 (nitrogen top-dressing. Total mixture crude proteins yields were from 845 kg ha-1 (control up to 998 kg ha-1 (nitrogen top-dressing.

  16. Effect of inoculation and nitrogen top-dressing on yields and fodder value of winter pea cv. Maksimirski ozimi in wheat cv. Sana mixture

    Directory of Open Access Journals (Sweden)

    Darko Uher

    2007-06-01

    Full Text Available Two year field trials (1999 - 2001 were performed to determine theinoculation seed winter pea and nitrogen top-dressing effect on number and active nodules of pea root and also on the green mass and dry matter yield of winter pea cv. Maksimirski ozimi and wheat cv. Sana mixture. Immediately before sowing the inoculation of pea seeds was accomplished by the indigenous variety of Rhizobium leguminosarum bv. viciae which belongs to the collection of Department of Microbiology at the Faculty of Agriculture University of Zagreb. The highest total nodule number on pea root (159 was determined on the inoculated variant as well as active nodule 144. Average mixture green mass yield were ranging from 24,65 t ha-1 (control up to 35,50 t ha-1 (inoculation. Total dry matter yields were ranging from 3,93 t ha-1 (control up to 5,66 t ha-1 (inoculation. Yields crude proteins pea in 2001 were ranging from 692 kg ha-1 (control up to 1058 kg ha-1 (inoculation and for wheat, those values ranged from 199 kg ha-1 (control up to 454 kg ha-1 (nitrogen top-dressing. Total crude proteins mixture yields were in range from 891 kg ha-1 (control up to 1360 kg ha-1 (inoculation.

  17. Analysis of rhizobial endosymbionts of Vicia, Lathyrus and Trifolium species used to maintain mountain firewalls in Sierra Nevada National Park (South Spain).

    Science.gov (United States)

    Villadas, Pablo J; Lasa, Ana V; Martínez-Hidalgo, Pilar; Flores-Félix, José David; Martínez-Molina, Eustoquio; Toro, Nicolás; Velázquez, Encarna; Fernández-López, Manuel

    2017-03-01

    Forest fires lead to the annual disappearance of many natural formations that require the creation of firewall areas. They can be maintained by enriching their pastures with attractive plants for grazing livestock, mainly legumes, which have a high protein content and low dependence on N fertilizers due to their ability to establish nitrogen-fixing symbiosis with rhizobia. In this study, the rhizobia isolated from the nodules of six legumes from the genera Vicia, Lathyrus and Trifolium were analysed in a firewall zone established in Lanjarón (Granada) close to the Sierra Nevada National Park (Spain). The results showed a high genetic diversity of the isolated strains that had 3, 16, 14 and 13 different types of rrs, recA, atpD and glnII genes, respectively. All strains were phylogenetically close to the species from the Rhizobium leguminosarum group, although they were not identified as any of them. The isolated strains belonged to the symbiovars viciae and trifolii but high phylogenetic diversity was found within both symbiovars, since there were 16 and 14 nodC gene types, respectively. Some of these strains clustered with strains isolated in other countries and continents, but others formed atpD, recA, glnII and nodC clusters and lineages only found to date in this study. Copyright © 2016 Elsevier GmbH. All rights reserved.

  18. Symbiont effect of Rhizobium bacteria and vesicular arbuscular mycorrhizal fungi on Pisum sativum in recultivated mine spoils

    Energy Technology Data Exchange (ETDEWEB)

    Biro, B.; Voros, I.; Kovespechy, K.; Szegi, J. (Hungarian Academy of Sciences, Budapest (Hungary). Soil Science and Agricultural Chemistry Research Institute)

    The frequency (F%) of spontaneous endomycorrhizal fungal infection (AMF) and the root modulation of Pisum sativum L, was studied after 8 and 15 years of recultivation in 4 soils (andesitic tuff yellow sand, yellow clay, and grey clay) disturbed by mining activities. The effects of Rhizobium inoculation and the interaction of both microsymbionts with plant production were also examined along with humus content and the humus stability coefficient, in the following variations: control, NPK fertilizer, NPK+lignite, NPK+straw and sewage sludge. Dump spoils originating from deep geological layers were poor in organic materials. After 12 years of recultivation, the humus content increased significantly. No such increase was noted in grey clay and the natural, brown forest soil used as an undisturbed (control) sample. The degradation of soils by mining brings about a decrease in the rhizobial and mycorrhizal population, so the number of spontaneous Rhizobium nodules is relatively low and does not influence the yield of peas. Inoculation with a selected effective strain of Rhizobium leguminosarum bv. viceae, however, enhanced dry matter production in these microbiologically degraded soils. Although the frequency of AMF infection was also higher after rhizobial inoculation, no positive correlation was found between dry matter production of peas and the F% of AMF. Spoils treated with lignite, straw, and sewage sludge had an especially high level of spontaneous mycorrhizal populations, indicating that organic materials had a favorable effect on the recultivation processes and fertility of mine spoils.

  19. Diversity and characterization of ramie-degumming strains

    Directory of Open Access Journals (Sweden)

    Shengwen Duan

    2012-04-01

    Full Text Available Ramie (Boehmeria nivea and Boehmeria tenacissima is a widely used fiber crop. Traditional water retting or chemical boiling method performed in order to extract ramie fiber seriously pollute the environment and severely damage the fiber, so biological method is the general trend of the fiber-extracting industry. Some strains (687, involving 26 genera and 43 species, were collected from the three samples, which produce hydrolyzed circles in the selective culture medium in order to detect the degumming effect and to compare the enzyme activity. Among these strains, 13 of them did not produce cellulase and had a ramie decreasing weight rate above 25 %, which were regarded as efficient ramie-degumming strains named from R1 to R13. R1 to R13 belonged to Amycolata autotrobutylicun, Bacillus subtilis, Clostridium acetobutylicum, Bacillus subtilis, Rhizobium leguminosarum, Bacteroides finegoldii, Streptomyces lividans, Bacillus amyloliquefaciens, Clostridium acetobutylicum, Pseudomonas brassicacearum, Bacillus pumilus, Bacillus licheniformis, Pectobacterium wasabiae respectively. Bacteroides sp., Rhizobium sp. and Pseudomonas sp. were firstly reported to be used in ramie-degumming. At the same time, the pectinase was the key enzyme in the ramie-degumming process.

  20. Ratiometric Gas Reporting: A Nondisruptive Approach To Monitor Gene Expression in Soils.

    Science.gov (United States)

    Cheng, Hsiao-Ying; Masiello, Caroline A; Del Valle, Ilenne; Gao, Xiaodong; Bennett, George N; Silberg, Jonathan J

    2018-03-16

    Fluorescent proteins are ubiquitous tools that are used to monitor the dynamic functions of natural and synthetic genetic circuits. However, these visual reporters can only be used in transparent settings, a limitation that complicates nondisruptive measurements of gene expression within many matrices, such as soils and sediments. We describe a new ratiometric gas reporting method for nondisruptively monitoring gene expression within hard-to-image environmental matrices. With this approach, C 2 H 4 is continuously synthesized by ethylene forming enzyme to provide information on viable cell number, and CH 3 Br is conditionally synthesized by placing a methyl halide transferase gene under the control of a conditional promoter. We show that ratiometric gas reporting enables the creation of Escherichia coli biosensors that report on acylhomoserine lactone (AHL) autoinducers used for quorum sensing by Gram-negative bacteria. Using these biosensors, we find that an agricultural soil decreases the bioavailable concentration of a long-chain AHL up to 100-fold. We also demonstrate that these biosensors can be used in soil to nondisruptively monitor AHLs synthesized by Rhizobium leguminosarum and degraded by Bacillus thuringiensis. Finally, we show that this new reporting approach can be used in Shewanella oneidensis, a bacterium that lives in sediments.

  1. Importance of winter pea cv. Maksimirski ozimi in production of the milk on family farms in Croatia

    Directory of Open Access Journals (Sweden)

    Darko Uher

    2008-12-01

    Full Text Available Two year field trials (1999-2001 were carried out to determine the effect of seed winter pea inoculation and nitrogen top-dressing on number and nodule dry weight g/plant of pea root and also on the yield of winter pea cv. Maksimirski ozimi and triticale cv. Clercal mixture. Just before sowing the inoculation of pea seeds was performed by the variety of Rhizobium leguminosarum bv. viciae which is part of the microbial collection of the Department of Microbiology at the Faculty of Agriculture University of Zagreb. The highest total nodule number on pea root (28 nodule/plant was determined on the inoculated variant 2 as well as nodule dry weight (0,175 g/plant. Average pea seed yield were ranging from 1327 kg ha-1 (control up to 1825 kg ha-1 (inoculation. Average triticale grain yield were ranging from 2375 kg ha-1 (control up to 3345 kg ha-1 (nitrogen top-dressing. Average total grain yield of winter peas in mixture triticale were ranging from 3702 kg ha-1 (control up to 5045 kg ha-1 (nitrogen top-dressing. This paper and given results are a humble contribution to the research of pea growth in the Republic of Croatia.

  2. Symbiotic effectiveness and phylogeny of rhizobia isolated from faba bean (Vicia faba L.) in Sichuan hilly areas, China.

    Science.gov (United States)

    Xu, Kai Wei; Zou, Lan; Penttinen, Petri; Wang, Ke; Heng, Nan Nan; Zhang, Xiao Ping; Chen, Qiang; Zhao, Ke; Chen, Yuan Xue

    2015-10-01

    A total of 54 rhizobial strains were isolated from faba bean root nodules in 21 counties of Sichuan hilly areas in China, and their symbiotic effectiveness, genetic diversity and phylogeny were assessed. Only six strains increased the shoot dry mass of the host plant significantly (P ≤ 0.05). Based on the cluster analysis of combined 16S rDNA and intergenic spacer region (IGS) PCR-RFLP, the strains were divided into 31 genotypes in 11 groups, indicating a high degree of genetic diversity among the strains. The sequence analysis of three housekeeping genes (atpD, glnII and recA) and 16S rDNA indicated that the strains represented two R. leguminosarum, two Rhizobium spp., R. mesosinicum, Agrobacterium sp. and A. tumefaciens. The strains representing four Rhizobium species were divided into two distinct nodC and nifH genotypes. However, the phylogeny of housekeeping genes and symbiotic genes was not congruent, implying that the strains had been shaped by vertical evolution of the housekeeping genes and lateral evolution of the symbiotic genes. Copyright © 2015 Elsevier GmbH. All rights reserved.

  3. Infestation of Phaseolus Vulgaris (L) by the beanfly Ophiomyia Spp. (Diptera: Agromyzidae) and Its Management by Cultural Practices

    International Nuclear Information System (INIS)

    Kayitare, Joseph Sibomana

    1993-04-01

    weed-free, and weeding three weeks after plant emergence, provided better grain yield. Plant spacing studies showed an increase of bean fly infestation in wider spacings. The optimal plant density of 222,222/ha (30x15 cm) seemed to give the least bean fly infestation and a high grain yield. Early planting with the first rains reduced bean fly infestation and gave better grain yield. Two species of bean fly, Ophiomyia phaseoli Tryon and O. spencerella Greathead, were identified in Oyugis, where O. Spencerella was more predominant. Parasitoids regulated bean fly population in nature. Six species of parasitoids were identified, with Opius phaseoli being the dominant species. They were host density-dependent and reduced bean fly population by 14-24%, The experimental results indicate that the use of nitrogen and phosphorus fertilizers, mono cropping system, optimal plant density of 222,222 plants/ha and early planting time were good cultural practices that can form a part of an integrated pest management (IPM) strategy to reduce bean fly infestation with consequent increase in bean production. (au)

  4. Severidade da mela da soja causada por Rhizoctonia solani AG-1 IA em função de doses de potássio Severity of hte foliar blight of the soylean caused by Rhizoctonia solani AG-1 IA infunction of doses of potassium

    Directory of Open Access Journals (Sweden)

    Marco Antonio Basseto

    2007-03-01

    Full Text Available O fungo Rhizoctonia solani pertencente ao grupo de anastomose 1 IA (AG-1 IA é um dos patógenos mais importantes afetando a cultura da soja no Brasil. Este fungo causa queima da folha e/ou mela em soja, para a qual medidas de manejo cultural são consideradas alternativas importantes para controle antes do estabelecimento da doença. Há evidências de que a adubação potássica diminui substancialmente a severidade dos sintomas de várias doenças da soja como a queima foliar (Cercospora kikuchii, a seca da haste e da vagem (Phomopsis phaseoli var. sojae e o cancro da haste (Diaporthe phaseolorum f. sp. meridionalis. Apesar das evidências do efeito do potássio no controle de várias doenças da soja, não há informação na literatura sobre o efeito desse nutriente no controle da mela. A hipótese testada foi que a mela da soja pode ser controlada através de incrementos na adubação potássica. De maneira geral, concluiu-se que, sob condições de casa de vegetação, o incremento de K no solo não resultou no controle da mela da soja. É necessário, entretanto, confirmar esta observação conduzindo-se experimentos sob condições de campo, podendo-se incluir a avaliação do efeito da doença sob aspectos da produção.The fungus Rhizoctonia solani, belonging to anastomosis group 1IA (AG-1 IA is one of the most important pathogens affecting soybean in Brazil. This fungus causes aerial or foliar blight of soybean, and cultural measures are thought as important choices for the control before the establishment of the disease. Based on evidences that potassium amendments can substantially reduce the severity of several soybean diseases such as Cercospora leaf blight (Cercospora kikuchii, pod and stem blight (Phomopsis phaseoli var. sojae and stem canker (Diaporthe phaseolorum f. sp. meridionalis. Despite all evidence, there is no information in the literature about the effect of potassium controlling the soybean foliar blight. The

  5. Métodos para la detección de Agrobacterium a partir de muestras de material vegetal, suelo y agua Methods for the detection of Agrobacterium from plant, soil and water samples

    Directory of Open Access Journals (Sweden)

    Adriana M Alippi

    2011-12-01

    Full Text Available El género Agrobacterium incluye especies ftopatógenas que inducen la formación de agallas en el cuello o la proliferación de raíces en cabellera en más de 600 especies de dicotiledóneas, y especies no patógenas cuyo hábitat natural es el suelo. Como no es posible erradicar a las especies patógenas y habida cuenta de que más del 80 % de las infecciones puede provenir de viveros, es importante evitar la diseminación de la enfermedad. Por ello, el objetivo de este trabajo ha sido desarrollar técnicas sensibles y precisas que, aisladamente o combinadas, permitan detectar la presencia de especies y biovares de Agrobacterium a partir de muestras de material vegetal, suelo y agua. Se comprobó que con la estrategia combinada de realizar aislamientos en los medios semiselectivos D1, D1-M y YEM-RCT; PCR multiplex sobre el gen 23S ADNr; PCR específca sobre los genes virC1 y virC2 y bioensayos en plántulas de pimiento cv. California Wonder y en hojas cortadas de kalanchoe, se reduce la posibilidad de obtener falsos negativos y/o falsos positivos. Por lo expuesto, esta combinación de técnicas constituye una herramienta adecuada para el diagnóstico de cepas patógenas de Agrobacterium a partir de distintos tipos de muestras.The genus Agrobacterium includes phytopathogenic bacteria that induce the development of root crown galls and/or aerial galls at the base of the stem or hairy roots on more than 600 species of plants belonging to 90 dicotyledonous families and non-pathogenic species. These bacteria being natural soil inhabitants are particularly diffcult to eradicate, which is a problem in nurseries where more than 80% of infections occur. Since early detection is crucial to avoid the inadvertent spread of the disease, the aim of this work was to develop sensitive and precise identifcation techniques by using a set of semi-selective and differential culture media in combination with a specifc PCR to amplify a partial sequence derived

  6. Mutation breeding in Brazil

    Energy Technology Data Exchange (ETDEWEB)

    Neto, A T; Menten, J O.M. [Centro de Energia Nuclear na Agricultura, Piracicaba (Brazil); Ando, A

    1980-03-01

    How mutation induction is used for plant breeding in Brazil is reported. For upland rice, the combined treatment with gamma-ray and mutagens (ethylene imine or ethylmethane sulfonate) has been used on the variety, Dourado Precoce, and some mutants with shortculm length and/or earliness without altering the productivity have been obtained. A project on the quantitative and qualitative protein improvement in upland rice was also started in 1979. In corn, the effect of gamma-irradiation on heterosis has been analyzed, and it was found that the single hybrids from two parental lines derived from irradiated seeds had increased ear productivity. For beans (Phaseolus yulgaris), gamma-irradiation and chemical mutagens have been used to induce the mutants with different seed color, disease resistance to golden mosaic virus and Xanthomonas phaseoli, earliness, high productivity and high protein content. Some mutants with partly improved characters have been obtained in these experiments. Two varieties of wheat tolerant to aluminum toxicity have been obtained, but the one showed high lodging due to its unfavorable plant height, and the other was highly susceptible to culm rust. Therefore, irradiation experiments have been started to improve these characters. The projects involving the use of gamma-irradiation have been tested to obtain the mutant lines insensitive to photoperiod and resistant to bud-blight in soybean, the mutant lines resistant to mosaic virus in papaya, the photoperiod-insensitive mutants in sorghum, the mosaic virus resistant and non-flowering mutants in sugar cane, and the Fusarium and nematode-resistant mutants in black pepper.

  7. MOLECULAR CHARACTERIZATION AND IDENTIFICATION OF BIOCONTROL ISOLATES OF TRICHODERMA HARZIANUM FROM EMBU DISTRICT, KENYA

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    Elizabeth M Siameto

    2010-10-01

    Full Text Available Species in the genus Trichoderma are important as commercial source of several enzymes, biofungicides, and growth promoters. The most common biological control agents of the genus are strains of T. harzianum, T .viride and T. viriens. In this study, sixteen selected isolates of T. harzianum from different land use types in Embu, Kenya were tested for anatogonistic action against five soil borne phytopathogenic fungi (Rhizoctonia solani, Pythium sp, Fusarium graminearum, F. oxysporum f. sp phaseoli and F. oxysporum f. sp Lycopersici using dual culture assay and through production of non-volatile inhibitors. Seven isolates were further characterized using RAPD-PCR procedure to determine genetic variability. All T. harzianum isolates had considerable antagonistic effect on mycelial growth of the pathogens in dual cultures compared to the controls. Maximum inhibition occurred in Pythium sp-055E interaction (73%.The culture filtrates obtained from Czapek’s liquid medium reduced the dry weight (mg of the mycelia significantly while those from the potato dextrose broth showed minimum inhibition growth. Pythium sp was inhibited the most compared to other pathogens. Genetic similarities generated using Jaccard’s coefficient of similarity ranged from 0.231 between isolates 055E and 011E to 0.857 between isolates 010E and 015E. The technique of RAPD was efficient in demonstrating the DNA polymorphism in the isolates of T. harzianum tested showing intraspecific genetic variability. Since all T. harzianum isolates evaluated were effective in controlling colony growth of the soil borne pathogens both in dual cultures and in culture filtrates they should be tried as a broad spectrum biological control agent in the greenhouse and under field conditions.

  8. The FTF gene family regulates virulence and expression of SIX effectors in Fusarium oxysporum.

    Science.gov (United States)

    Niño-Sánchez, Jonathan; Casado-Del Castillo, Virginia; Tello, Vega; De Vega-Bartol, José J; Ramos, Brisa; Sukno, Serenella A; Díaz Mínguez, José María

    2016-09-01

    The FTF (Fusarium transcription factor) gene family comprises a single copy gene, FTF2, which is present in all the filamentous ascomycetes analysed, and several copies of a close relative, FTF1, which is exclusive to Fusarium oxysporum. An RNA-mediated gene silencing system was developed to target mRNA produced by all the FTF genes, and tested in two formae speciales: F. oxysporum f. sp. phaseoli (whose host is common bean) and F. oxysporum f. sp. lycopersici (whose host is tomato). Quantification of the mRNA levels showed knockdown of FTF1 and FTF2 in randomly isolated transformants of both formae speciales. The attenuation of FTF expression resulted in a marked reduction in virulence, a reduced expression of several SIX (Secreted In Xylem) genes, the best studied family of effectors in F. oxysporum, and lower levels of SGE1 (Six Gene Expression 1) mRNA, the presumptive regulator of SIX expression. Moreover, the knockdown mutants showed a pattern of colonization of the host plant similar to that displayed by strains devoid of FTF1 copies (weakly virulent strains). Gene knockout of FTF2 also resulted in a reduction in virulence, but to a lesser extent. These results demonstrate the role of the FTF gene expansion, mostly the FTF1 paralogues, as a regulator of virulence in F. oxysporum and suggest that the control of effector expression is the mechanism involved. © 2016 The Authors Molecular Plant Pathology Published by British Society for Plant Pathology and John Wiley & Sons Ltd.

  9. Microflora en semillas de frijol

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    Jos\\u00E9 B. Membre\\u00F1o

    2001-01-01

    Full Text Available Microflora en semillas de frijol (Phaseolus vulgaris L.. Se estudió la microflora bacteriana presente en semillas de frijol y su relación con Xanthomonas campestris pv. phaseoli (Xcp, en 118 genotipos procedentes de VIDAC-98, INTA- Nicaragua, TARS-USDA e Isabela-P.R. Se utilizaron cinco métodos de aislamiento: semilla desinfectada con hipoclorito de sodio, semilla en caldo nutritivo refrigerada por una hora, dispersión de 0,1 ml de suspensión de semillas en medio sólido, siembra líquida de 1 ml de suspensión y semilla en caldo nutritivo, agitado y refrigerado por 24 horas. Se aislaron 104 colonias amarillas de 41 genotipos. Treinta y seis colonias fueron KOH positivo (Gram negativo, 68 negativo (Gram positivo y 34 hidrolizaron almidón. Las colonias de pigmentación amarilla resultaron no patogénicas bajo condiciones de invernadero. Estas se identificaron con el sistema BIOLOG como: Pantoea agglomerans (25, Xanthomonas campestris (2, Enterobacter agglomerans (2, Sphingomonas paucimobilis (2, Pseudomonas fluorescens y Flavimonas oryzihabitans. En adición, los genotipos portaron colonias con pigmentación distinta a la amarilla. En las pruebas de antagonismo se identificaron colonias con actividad de deoxyribonucleasa y de antibiosis a Xcp. De éstas, 15 colonias inhibieron a Xcp significativamente. Se identificaron los hongos Rhizoctonia solani, Penicillium spp., Fusarium spp., Aspergillus flavus, Rhizopus nigricans y Macrophomina phaseolina en un 52,9 % del total de genotipos evaluados

  10. QTL and candidate genes associated with common bacterial blight resistance in the common bean cultivar Longyundou 5 from China

    Institute of Scientific and Technical Information of China (English)

    Jifeng Zhu; Jing Wu; Lanfen Wang; Matthew W. Blair; Zhendong Zhu; Shumin Wang

    2016-01-01

    Common bacterial blight (CBB), caused by Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans (Xff), is a worldwide disease of common bean (Phaseolus vulgaris L.). Longyundou 5, a Chinese cultivar in the Mesoamerican gene pool of common bean, displays resistance to the Xff strain XSC3-1. To identify the genetic mechanisms behind this resistance, we crossed Long 5 with a susceptible genotype to develop a mapping population of F2 plants. Plant resistance to CBB was identified at 14 and 21 days after inoculation with Xff strain XSC3-1. A major QTL at 14 and 21 days after inoculation was mapped on chromosome Pv10 with LOD scores of 6.41 and 5.35, respectively. This locus was associated with SAP6, a previously-identified and much-used dominant marker, but in a 4.2 cM interval between new codominant markers BMp10s174 and BMp10s244. Ten candidate genes were found between markers BMp10s174 and BMp10s244 on chromosome Pv10 and could encode defense response proteins responding to CBB pathogens. Four pairs each of epistatic QTL for CBB resistance were detected at 14 and 21 days after inoculation. Phenotypic variation explained by the epistatic QTL ranged from 7.19%to 12.15%and 7.72%to 8.80%at 14 and 21 days after inoculation, respectively. These results confirmed the importance of epistasis in CBB resistance in common bean. The adjacent markers found may be more efficient for marker assisted selection in common bean breeding for CBB resistance owing to their closer linkage to the target QTL.

  11. QTL and candidate genes associated with common bacterial blight resistance in the common bean cultivar Longyundou 5 from China

    Institute of Scientific and Technical Information of China (English)

    Jifeng; Zhu; Jing; Wu; Lanfen; Wang; Matthew; W.Blair; Zhendong; Zhu; Shumin; Wang

    2016-01-01

    Common bacterial blight(CBB), caused by Xanthomonas axonopodis pv. phaseoli and Xanthomonas fuscans subsp. fuscans(Xff), is a worldwide disease of common bean(Phaseolus vulgaris L.).Longyundou 5, a Chinese cultivar in the Mesoamerican gene pool of common bean, displays resistance to the Xff strain XSC3-1. To identify the genetic mechanisms behind this resistance,we crossed Long 5 with a susceptible genotype to develop a mapping population of F2 plants.Plant resistance to CBB was identified at 14 and 21 days after inoculation with Xff strain XSC3-1.A major QTL at 14 and 21 days after inoculation was mapped on chromosome Pv10 with LOD scores of 6.41 and 5.35, respectively. This locus was associated with SAP6, a previouslyidentified and much-used dominant marker, but in a 4.2 cM interval between new codominant markers BMp10s174 and BMp10s244. Ten candidate genes were found between markers BMp10s174 and BMp10s244 on chromosome Pv10 and could encode defense response proteins responding to CBB pathogens. Four pairs each of epistatic QTL for CBB resistance were detected at 14 and 21 days after inoculation. Phenotypic variation explained by the epistatic QTL ranged from 7.19% to 12.15% and 7.72% to 8.80% at 14 and 21 days after inoculation, respectively. These results confirmed the importance of epistasis in CBB resistance in common bean. The adjacent markers found may be more efficient for marker assisted selection in common bean breeding for CBB resistance owing to their closer linkage to the target QTL.

  12. Verrucosispora sonchi sp. nov., a novel endophytic actinobacterium isolated from the leaves of common sowthistle (Sonchus oleraceus L.).

    Science.gov (United States)

    Ma, Zhaoxu; Zhao, Shanshan; Cao, Tingting; Liu, Chongxi; Huang, Ying; Gao, Yuhang; Yan, Kai; Xiang, Wensheng; Wang, Xiangjing

    2016-12-01

    A novel actinobacterium, designated strain NEAU-QY3T, was isolated from the leaves of Sonchus oleraceus L. and examined using a polyphasic taxonomic approach. The organism formed single spores with smooth surface on substrate mycelia. Phylogenetic analysis based on the 16S rRNA gene sequence indicated that the strain had a close association with the genus Verrucosispora and shared the highest sequence similarity with Verrucosispora qiuiae RtIII47T (99.17 %), an association that was supported by a bootstrap value of 94 % in the neighbour-joining tree and also recovered with the maximum-likelihood algorithm. The strain also showed high 16S rRNA gene sequence similarities to Xiangella phaseoli NEAU-J5T (98.78 %), Jishengella endophytica 202201T (98.51 %), Micromonospora eburnea LK2-10T (98.28 %), Verrucosispora lutea YIM 013T (98.23 %) and Salinispora pacifica CNR-114T (98.23 %). Furthermore, phylogenetic analysis based on the gyrB gene sequences supported the conclusion that strain NEAU-QY3T should be assigned to the genus Verrucosispora. However, the DNA-DNA hybridization relatedness values between strain NEAU-QY3T and V. qiuiae RtIII47T and V. lutea YIM 013T were below 70 %. With reference to phenotypic characteristics, phylogenetic data and DNA-DNA hybridization results, strain NEAU-QY3T was readily distinguished from its most closely related strains and classified as a new species, for which the name Verrucosispora sonchi sp. nov. is proposed. The type strain is NEAU-QY3T (=CGMCC 4.7312T=DSM 101530T).

  13. A Comparison of the Molecular Organization of Genomic Regions Associated with Resistance to Common Bacterial Blight in Two Phaseolus vulgaris Genotypes

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    Gregory E. Perry

    2013-08-01

    Full Text Available Resistance to common bacterial blight, caused by Xanthomonas axonopodis pv. phaseoli, in Phaseolus vulgaris is conditioned by several loci on different chromosomes. Previous studies with OAC-Rex, a CBB-resistant, white bean variety of Mesoamerican origin, identified two resistance loci associated with the molecular markers Pv-CTT001 and SU91, on chromosome 4 and 8, respectively. Resistance to CBB is assumed to be derived from an interspecific cross with Phaseolus acutifolius in the pedigree of OAC-Rex. Our current whole genome sequencing effort with OAC-Rex provided the opportunity to compare its genome in the regions associated with CBB resistance with the v1.0 release of the P. vulgaris line G19833, which is a large seeded bean of Andean origin, and (assumed to be CBB susceptible.. In addition, the genomic regions containing SAP6, a marker associated with P. vulgaris-derived CBB-resistance on chromosome 10, were compared. These analyses indicated that gene content was highly conserved between G19833 and OAC-Rex across the regions examined (>80%. However, fifty-nine genes unique to OAC Rex were identified, with resistance gene homologues making up the largest category (10 genes identified. Two unique genes in OAC-Rex located within the SU91 resistance QTL have homology to P. acutifolius ESTs and may be potential sources of CBB resistance. As the genomic sequence assembly of OAC-Rex is completed, we expect that further comparisons between it and the G19833 genome will lead to a greater understanding of CBB resistance in bean.

  14. Utilización del medio Mrs-s en el aislamiento de bacterias lácticas mesofilas en leche de cabra Utilización del medio Mrs-s en el aislamiento de bacterias lacticas mesofilas en leche de cabra

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    Fortes F. Celia L. de Luces

    1990-12-01

    Full Text Available Con el fin de evaluar el MRS-S (Sorbato al 0.10% en el aislamiento de bacterias lácticas se cultivaron muestras de leche de cabra cruda en MRS-S y PCA en profundidad y se incubaron en aerobiósis a 320C durante 48 horas. Los cocos gram positivos, catalasa negativos que crecieron en MRS-S se aislaron y sometieron a caracterización preliminar a través del crecimiento en agar MRS-S, MRS-T (tetraciclina 0.20 µg/ml, N-L (bacterias aromáticas, reducción de la leche tornasolada a 40 y 21oC y crecimiento a 45 y 10oC.Las cepas seleccionadas se sometieron a caracterización fisiológica y Bioquímica. El medio MRS-S mostró ser efectivo con un porcentaje de inhibición de la flora indeseable del 86.56%, y adecuado por el aislamiento de Lactococcus. De acuerdo con los perfiles taxonómicos se consiguió aislar de un total de 156 colonias dos Lactococcus lactis subsp. lactis y un Lactococcus lactis biovar diacetilactis.With the objective of evaluating the MRS-S (0.10% of sorbate in the isolation of lactic bacteria samples of raw goat milk were cultivated in MRS-S and PCA in deep and incubated in aerobic conditions for 48 hours al 32oC. Gram positive coccus, negative catalase which grew in MRS-S were isolated and preliminarly characterized through the growing process in agar MRS-S, MRS-T (O.20ltg/ml tetracycline, N-L (aromatic bacteria, litmus milk reduction at 40 and 210C and growing at 45 and 10oC. Selected strains were subject to the physiological and biochemical characterization. MRS-S media showed to be effective with an 86.56% of inhibition for indesirable bacteria and adequated for Lactococcus isolation. Related to taxonomic profiles from 156 colonies were isolated two lactococcus lactis subsp. lactis and one Lactococcus luctis biovar diacetilactis.

  15. Physicochemical and microbiological study of “shmen”, a traditional butter made from camel milk in the Sahara (Algeria: isolation and identification of lactic acid bacteria and yeasts

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    Mourad, Kacem

    2006-06-01

    Full Text Available Microorganisms (aerobic bacteria, coliforms, lactic acid bacteria, psychrotrophs, lipolytic bacteria and yeasts were isolated from 20 samples of shmen, a traditional clarified butter made from sour camel milk in the Algerian Sahara. The values of pH, titratable acidity, NaCl, total solid, moisture, and fat content ranged from : 3.11-4.97, 0.19-0.36%, 1.04-2.15%, 64.03-65.11%, 34.40-34.99%, and 49.90-56% respectively. A total of 181 isolates of lactic acid bacteria were identified as Lactobacillus plantarum (40 strains, Lactobacillus delbrueckii ssp. bulgaricus (35 strains, Lactococcus lactis ssp. lactis biovar diacetylacti (22 strains, Lactococcus lactis ssp. cremoris (18 strains, Lactobacillus paracasei ssp. paracasei (10 strains, Leuconostoc pseudomesenteroides (9 strains and Leuconostoc gelidum (12 strains Enterococcus faecium (35 strains. Yeasts were isolated from all samples (55 isolates. Of these, 40 were identified as Saccharomyces cerevisiae and 15 isolates were identified as Saccharomyces sp.Se aislaron los microorganismos (bacterias aeróbicas, coliformes, bacterias acido lácticas, bacterias lipolíticas y levaduras de 20 muestras de “shmen”, una matequilla tradicional del Sahara argelino hecha a partir de leche de camella. Los valores de pH, acidez, libre, Nacl, solidos totales, humedad y grasa oscilaron entre 3,11-4,97, 0,19-0,36%, 1.04-2,15%, 64,03-65,11%, 34,40-34,99% y 49,90-56,00%, respectivamente. Entre los 181 cultivos puros de bacterias lácticas se identificaron Lactobacillus plantarum (40 cepas, Lactobacillus delbrueckii ssp. bulgaricus (35 cepas, Lactococcus lactis ssp. lactis biovar diacetylacti (22 cepas, Lactococcus lactis ssp. cremoris (18 cepas, Lactobacillus paracasei ssp. paracasei (10 cepas, Leuconostoc pseudomesenteroides (9 cepas and Leuconostoc gelidum (12cepas Enterococcus faecium (35 cepas. Asimismo, se detectaron levaduras en todas las muestras (55 cultivos puros. De estos, 40 se identificaron como

  16. Tipificación molecular de Brucella abortus cepa 19 y su aplicación al control de biológicos Molecular characterization of Brucella abortus strain 19 and its application for controlling biologics

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    M.E. Pavan

    2005-09-01

    Full Text Available Brucella abortus es el agente etiológico de la brucelosis bovina. La cepa 19, utilizada en la elaboración de vacunas, puede ser identificada a través de una deleción en la región eri asociada con la sensibilidad al eritritol. Se optimizó un ensayo de PCR para caracterizar específicamente esta cepa. El método que describimos es un procedimiento rápido para identificar B. abortus y simultáneamente diferenciar la cepa 19 de otras cepas de B. abortus biovar 1. Hemos aplicado este ensayo para la detección de la cepa 19 en vacunas contra la brucelosis bovina elaboradas en Argentina. Los resultados indican que este método podría ser útil para el seguimiento de las cepas madres y semillas utilizadas en la producción industrial de esta vacuna. Esta metodología también contribuiría a la reducción del riesgo de la infección adquirida en el laboratorio y podría aplicarse como prueba de rutina para confirmar la presencia de B. abortus en vacunas no relacionadas.Brucella abortus is the etiological agent of bovine brucellosis. The strain 19 used in vaccine elaboration can be identified through a deletion in the eri region associated with its susceptibility to erythritol. We optimized a PCR assay for specific characterization of this strain. The method described here is a rapid procedure that enables identification of B. abortus, and simultaneous differentiation of the strain 19 from other B. abortus biovar 1 strains. We applied the assay to detect the strain 19 in vaccines against B. abortus produced in Argentina. The results show this method could be used to follow vaccine seed cultures of this strain. The methodology could also contribute to reduce the risk of a laboratory-acquired infection and could be of great help as a routine test for confirmation of B. abortus in non related vaccines.

  17. Yersinia pestis Caf1 Protein: Effect of Sequence Polymorphism on Intrinsic Disorder Propensity, Serological Cross-Reactivity and Cross-Protectivity of Isoforms.

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    Pavel Kh Kopylov

    Full Text Available Yersinia pestis Caf1 is a multifunctional protein responsible for antiphagocytic activity and is a key protective antigen. It is generally conserved between globally distributed Y. pestis strains, but Y. pestis subsp. microtus biovar caucasica strains circulating within populations of common voles in Georgia and Armenia were reported to carry a single substitution of alanine to serine. We investigated polymorphism of the Caf1 sequences among other Y. pestis subsp. microtus strains, which have a limited virulence in guinea pigs and in humans. Sequencing of caf1 genes from 119 Y. pestis strains belonging to different biovars within subsp. microtus showed that the Caf1 proteins exist in three isoforms, the global type Caf1NT1 (Ala48 Phe117, type Caf1NT2 (Ser48 Phe117 found in Transcaucasian-highland and Pre-Araks natural plague foci #4-7, and a novel Caf1NT3 type (Ala48 Val117 endemic in Dagestan-highland natural plague focus #39. Both minor types are the progenies of the global isoform. In this report, Caf1 polymorphism was analyzed by comparing predicted intrinsic disorder propensities and potential protein-protein interactivities of the three Caf1 isoforms. The analysis revealed that these properties of Caf1 protein are minimally affected by its polymorphism. All protein isoforms could be equally detected by an immunochromatography test for plague at the lowest protein concentration tested (1.0 ng/mL, which is the detection limit. When compared to the classic Caf1NT1 isoform, the endemic Caf1NT2 or Caf1NT3 had lower immunoreactivity in ELISA and lower indices of self- and cross-protection. Despite a visible reduction in cross-protection between all Caf1 isoforms, our data suggest that polymorphism in the caf1 gene may not allow the carriers of Caf1NT2 or Caf1NT3 variants escaping from the Caf1NT1-mediated immunity to plague in the case of a low-dose flea-borne infection.

  18. Prophage Rs551 and Its Repressor Gene orf14 Reduce Virulence and Increase Competitive Fitness of Its Ralstonia solanacearum Carrier Strain UW551.

    Science.gov (United States)

    Ahmad, Abdelmonim Ali; Stulberg, Michael J; Huang, Qi

    2017-01-01

    We previously characterized a filamentous lysogenic bacteriophage, ϕRs551, isolated directly from the race 3 biovar 2 phylotype IIB sequevar 1 strain UW551 of Ralstonia solanacearum grown under normal culture conditions. The genome of ϕRs551 was identified with 100% identity in the deposited genomes of 11 race 3 biovar 2 phylotype IIB sequevar 1 strains of R. solanacearum , indicating evolutionary and biological importance, and ORF14 of ϕRs551 was annotated as a putative type-2 repressor. In this study, we determined the effect of the prophage and its ORF14 on the virulence and competitive fitness of its carrier strain UW551 by deleting the orf14 gene only (the UW551 orf14 mutant), and nine of the prophage's 14 genes including orf14 and six out of seven structural genes (the UW551 prophage mutant), respectively, from the genome of UW551. The two mutants were increased in extracellular polysaccharide production, twitching motility, expression of targeted virulence and virulence regulatory genes ( pilT, egl, pehC, hrPB, and phcA ), and virulence, suggesting that the virulence of UW551 was negatively regulated by ϕRs551, at least partially through ORF14. Interestingly, we found that the wt ϕRs551-carrying strain UW551 of R. solanacearum significantly outcompeted the wt strain RUN302 which lacks the prophage in tomato plants co-inoculated with the two strains. When each of the two mutant strains was co-inoculated with RUN302, however, the mutants were significantly out-competed by RUN302 for the same colonization site. Our results suggest that ecologically, ϕRs551 may play an important role by regulating the virulence of and offering a competitive fitness advantage to its carrier bacterial strain for persistence of the bacterium in the environment, which in turn prolongs the symbiotic relationship between the phage ϕRs551 and the R. solanacearum strain UW551. Our study is the first toward a better understanding of the co-existence between a lysogenic phage and

  19. Volatile Organic Compounds in Naturally Fermented Milk and Milk Fermented Using Yeasts, Lactic Acid Bacteria and Their Combinations As Starter Cultures

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    Bennie C. Viljoen

    2007-01-01

    Full Text Available The volatile organic compounds present in 18 Zimbabwean naturally fermented milk (amasi samples and those produced by various yeasts, lactic acid bacteria (LAB and yeast/ LAB combinations were determined using headspace gas chromatography. The yeast strains used were: Candida kefyr 23, C. lipolytica 57, Saccharomyces cerevisiae 71, C. lusitaniae 68, C. tropicalis 78, C. lusitaniae 63, C. colliculosa 41, S. dairenensis 32, and Dekkera bruxellensis 43, and were coded Y1 to Y9, respectively. The LAB strains used were Lactococcus lactis subsp. lactis Lc39, L. lactis subsp. lactis Lc261, Lactobacillus paracasei Lb11, and L. lactis subsp. lactis biovar. diacetylactis C1, and were coded B1 to B4, respectively. Some of the volatile organic compounds found in amasi were acetaldehyde, ethanol, acetone, 2-methyl propanal, 2-methyl-1-propanol and 3-methyl-1-butanol. However, the levels of volatile organic compounds in the naturally fermented milk (NFM samples varied from one sample to another, with acetaldehyde ranging from 0.1–18.4 ppm, 3-methyl butanal from <0.1–0.47 ppm and ethanol from 39.3–656 ppm. The LAB/C. kefyr 23 (B/Y1 co-cultures produced significantly (p<0.05 higher levels of acetaldehyde and ethanol than the levels found in the NFM. The acetaldehyde levels in the B/Y1 samples ranged from 26.7–87.7 ppm, with L. lactis subsp. lactis biovar. diacetylactis C1 (B4 producing the highest level of acetaldehyde in combination with C. kefyr 23 (Y1. Using principal component analysis (PCA, most of the NFM samples were grouped together with single and co-cultures of Lc261, Lb11 and the non-lactose fermenting yeasts, mainly because of the low levels of ethanol and similar levels of 3-methyl butanal. Chromatograms of amasi showed prominent peak of methyl aldehydes and their alcohols including 3-methyl-butanal and 3-methyl-butanol, suggesting that these compounds are important attributes of Zimbabwean naturally fermented milk.

  20. Prophage Rs551 and Its Repressor Gene orf14 Reduce Virulence and Increase Competitive Fitness of Its Ralstonia solanacearum Carrier Strain UW551

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    Abdelmonim Ali Ahmad

    2017-12-01

    Full Text Available We previously characterized a filamentous lysogenic bacteriophage, ϕRs551, isolated directly from the race 3 biovar 2 phylotype IIB sequevar 1 strain UW551 of Ralstonia solanacearum grown under normal culture conditions. The genome of ϕRs551 was identified with 100% identity in the deposited genomes of 11 race 3 biovar 2 phylotype IIB sequevar 1 strains of R. solanacearum, indicating evolutionary and biological importance, and ORF14 of ϕRs551 was annotated as a putative type-2 repressor. In this study, we determined the effect of the prophage and its ORF14 on the virulence and competitive fitness of its carrier strain UW551 by deleting the orf14 gene only (the UW551 orf14 mutant, and nine of the prophage’s 14 genes including orf14 and six out of seven structural genes (the UW551 prophage mutant, respectively, from the genome of UW551. The two mutants were increased in extracellular polysaccharide production, twitching motility, expression of targeted virulence and virulence regulatory genes (pilT, egl, pehC, hrPB, and phcA, and virulence, suggesting that the virulence of UW551 was negatively regulated by ϕRs551, at least partially through ORF14. Interestingly, we found that the wt ϕRs551-carrying strain UW551 of R. solanacearum significantly outcompeted the wt strain RUN302 which lacks the prophage in tomato plants co-inoculated with the two strains. When each of the two mutant strains was co-inoculated with RUN302, however, the mutants were significantly out-competed by RUN302 for the same colonization site. Our results suggest that ecologically, ϕRs551 may play an important role by regulating the virulence of and offering a competitive fitness advantage to its carrier bacterial strain for persistence of the bacterium in the environment, which in turn prolongs the symbiotic relationship between the phage ϕRs551 and the R. solanacearum strain UW551. Our study is the first toward a better understanding of the co-existence between a

  1. Prophage Rs551 and Its Repressor Gene orf14 Reduce Virulence and Increase Competitive Fitness of Its Ralstonia solanacearum Carrier Strain UW551

    Science.gov (United States)

    Ahmad, Abdelmonim Ali; Stulberg, Michael J.; Huang, Qi

    2017-01-01

    We previously characterized a filamentous lysogenic bacteriophage, ϕRs551, isolated directly from the race 3 biovar 2 phylotype IIB sequevar 1 strain UW551 of Ralstonia solanacearum grown under normal culture conditions. The genome of ϕRs551 was identified with 100% identity in the deposited genomes of 11 race 3 biovar 2 phylotype IIB sequevar 1 strains of R. solanacearum, indicating evolutionary and biological importance, and ORF14 of ϕRs551 was annotated as a putative type-2 repressor. In this study, we determined the effect of the prophage and its ORF14 on the virulence and competitive fitness of its carrier strain UW551 by deleting the orf14 gene only (the UW551 orf14 mutant), and nine of the prophage’s 14 genes including orf14 and six out of seven structural genes (the UW551 prophage mutant), respectively, from the genome of UW551. The two mutants were increased in extracellular polysaccharide production, twitching motility, expression of targeted virulence and virulence regulatory genes (pilT, egl, pehC, hrPB, and phcA), and virulence, suggesting that the virulence of UW551 was negatively regulated by ϕRs551, at least partially through ORF14. Interestingly, we found that the wt ϕRs551-carrying strain UW551 of R. solanacearum significantly outcompeted the wt strain RUN302 which lacks the prophage in tomato plants co-inoculated with the two strains. When each of the two mutant strains was co-inoculated with RUN302, however, the mutants were significantly out-competed by RUN302 for the same colonization site. Our results suggest that ecologically, ϕRs551 may play an important role by regulating the virulence of and offering a competitive fitness advantage to its carrier bacterial strain for persistence of the bacterium in the environment, which in turn prolongs the symbiotic relationship between the phage ϕRs551 and the R. solanacearum strain UW551. Our study is the first toward a better understanding of the co-existence between a lysogenic phage and

  2. Determination of Molecular Genotyping of Ureaplasma SPP in Women with Genital Infections by 16S–23S rDNA PCR-RFLP Method

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    R. Mirnejad

    2011-04-01

    Full Text Available Introduction & Objective: So far, despite the wide range of methods such as analytic methods used for differentiation of Mycoplasma, the diagnosis of Mycoplasma species is still difficult. Generally the low-level discriminatory power of serological methods because of the rapid changes in size and phase of the dominant antigens in the immune cell surface of Mycoplasmas greatly limits their applicability to the typing of Mycoplasmas. On the contrary,molecular methods do not suffer from these drawbacks and can be used for typing of Mycoplasmas. The aim of this investigation was molecular identification and genotyping of ureaplasma SPP in women with genital infections by 16S–23S rDNA PCR-RFLP.Materials & Methods: Genital swabs were taken from 210 patients who referred to gynecology clinic of Rasool hospital in Tehran, Iran during December 2007 until June 2008. The swabs suspended in PBS, were immediately transferred to laboratory .Following DNA extraction, PCR assay was performed using a genus specific primer pair. These primer sets amplified a 559 bp fragment for Ureaplasma Spp. Samples containing bands of the expected size for Ureaplasma strains were subjected to digestion with different restriction endonuclease enzymes (AluI, Taq I, CacI8, BbsI, EcoRI. Results: Of the 210 samples, Ureaplasma Spp was isolated from 93 patients (44.3% by PCR and 69 samples by culture. In the present study only Biovar 1 (Ureaplasma parvum was isolated from clinical specimens and the results were confirmed using a cutting enzyme TaqI (enzyme specific species of ureaplasma SPP. The results of this analysis using PCR-RFLP and sequencing showed that all had the same genotype and shared identical sequence with the genome sequence of serovar 3 Ureaplasma parvum.Conclusion: Ureaplasma parvum is generally isolated from the genital samples. In this study all isolates were identical and no difference was found among the enzyme patterns of the bacteria after PCR-RFLP .So

  3. Efeito do boro na nodulação da ervilha cultivada em solos de várzea Boron effects on nodule development in pea plants cultivated in lowland soils

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    Watson Rogério de Azevedo

    2002-08-01

    Full Text Available O objetivo deste trabalho foi avaliar o efeito do boro na nodulação da ervilha (Pisum sativum L. cv. Torta de Flor Roxa, cultivada em solos de várzea em condições de casa de vegetação, entre maio e julho de 1998. Amostras de Neossolo Flúvico, Gleissolo Háplico, Gleissolo Melânico e Organossolo Mésico artificialmente drenado foram coletadas na camada de 0-20 cm. Essas amostras foram previamente cultivadas com rabanete, e receberam calcário, P, K, S, Cu e Zn e seis doses de B (0,0, 0,25, 0,5, 1,5, 3,0 e 6,0 mg dm-3. Por ocasião do cultivo da ervilha aplicou-se apenas K (100 mg dm-3. O delineamento experimental foi inteiramente casualizado, em esquema fatorial 4x6, com três repetições. Foram cultivadas três plantas por vaso, cujas sementes, na época da semeadura, foram submetidas à inoculação com estirpes de Rhizobium leguminosarum (BR 618 e BR 619. Para verificação da existência de estirpes nativas de rizóbio, cultivaram-se sem inoculação nas sementes, três vasos, de cada solo, que haviam recebido a mesma dose de calcário, a mesma adubação básica e 0,5 mg dm-3 de boro. Aos 45 dias após o plantio, em pleno florescimento, colheu-se o experimento. Não foram observadas estirpes nativas nos solos avaliados. Nos solos Gleissolo Háplico e Organossolo Mésico artificialmente drenado, as doses de B influenciaram a nodulação e a atividade da nitrogenase.The objective of this work was to evaluate the effect of boron on nodule development in pea Pisum sativum L. plants cultivated in lowland soils under greenhouse conditions from May to July, 1998. Samples of Alluvial Soil, Low Humic Gley, Humic Gley and artificially drained Bog Soil were collected from a layer 0-20 cm deep. The samples were previously cultivated with radish plants that received lime, P, K, S, Cu and Zn and doses of B (0, 0.25, 0.5, 1.5, 3.0 and 6.0 mg dm-3. Before pea cultivation only K was applied (100 mg dm-3. The experimental design was totally

  4. Genetic and phenotypic diversity of Rhizobium isolates from Southern Ecuador

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    Roldán Torres-Gutiérrez

    Full Text Available ABSTRACT Rhizobium-legume symbioses play relevant roles in agriculture but have not been well studied in Ecuador. The aim of this study was to characterize the genetic and phenotypic diversity of Rhizobium isolates associated with Phaseolus vulgaris from southern Ecuador. Morpho-cultural characterization, biochemical tests and physiological analyses were conducted to authenticate and determine the diversity of bacteria Rhizobium-like isolates. The genetic diversity of the isolates was determined by molecular techniques, which consisted of bacteria DNA extraction and amplification and sequencing of the 16S rRNA gene. The nodulation parameters and nitrogen fixation for P. vulgaris under greenhouse conditions were also assessed to determine the phenotypic diversity among isolates. Furthermore, bacteria indole-acetic-acid production was evaluated by the colorimetric method. Morpho-cultural and biochemical characteristic assessments demonstrated that Rhizobium-like bacteria was associated with the P. vulgaris nodules. The diversity among the isolates, as determined by physiological analyses, revealed the potential of several isolates to grow at different pH values, salinity conditions and temperatures. Partial sequencing of the 16S rRNA gene identified the Rhizobium genus in every sampling site. From a total of 20 aligned sequences, nine species of Rhizobium were identified. Nodule formation and biomass, as well as nitrogen fixation, showed an increase in plant phenotypic parameters, which could be influenced by IAA production, especially for the strains R. mesoamericanum NAM1 and R. leguminosarum bv. viciae COL6. These results demonstrated the efficiency of native symbiotic diazotrophic strains inoculants for legume production. This work can serve as the basis for additional studies of native Rhizobium strains and to help spread the use of biofertilizers in Ecuadorian fields.

  5. Candidatus Frankia Datiscae Dg1, the Actinobacterial Microsymbiont of Datisca glomerata, Expresses the Canonical nod Genes nodABC in Symbiosis with Its Host Plant

    Science.gov (United States)

    Persson, Tomas; Battenberg, Kai; Demina, Irina V.; Vigil-Stenman, Theoden; Vanden Heuvel, Brian; Pujic, Petar; Facciotti, Marc T.; Wilbanks, Elizabeth G.; O'Brien, Anna; Fournier, Pascale; Cruz Hernandez, Maria Antonia; Mendoza Herrera, Alberto; Médigue, Claudine; Normand, Philippe; Pawlowski, Katharina; Berry, Alison M.

    2015-01-01

    Frankia strains are nitrogen-fixing soil actinobacteria that can form root symbioses with actinorhizal plants. Phylogenetically, symbiotic frankiae can be divided into three clusters, and this division also corresponds to host specificity groups. The strains of cluster II which form symbioses with actinorhizal Rosales and Cucurbitales, thus displaying a broad host range, show suprisingly low genetic diversity and to date can not be cultured. The genome of the first representative of this cluster, Candidatus Frankia datiscae Dg1 (Dg1), a microsymbiont of Datisca glomerata, was recently sequenced. A phylogenetic analysis of 50 different housekeeping genes of Dg1 and three published Frankia genomes showed that cluster II is basal among the symbiotic Frankia clusters. Detailed analysis showed that nodules of D. glomerata, independent of the origin of the inoculum, contain several closely related cluster II Frankia operational taxonomic units. Actinorhizal plants and legumes both belong to the nitrogen-fixing plant clade, and bacterial signaling in both groups involves the common symbiotic pathway also used by arbuscular mycorrhizal fungi. However, so far, no molecules resembling rhizobial Nod factors could be isolated from Frankia cultures. Alone among Frankia genomes available to date, the genome of Dg1 contains the canonical nod genes nodA, nodB and nodC known from rhizobia, and these genes are arranged in two operons which are expressed in D. glomerata nodules. Furthermore, Frankia Dg1 nodC was able to partially complement a Rhizobium leguminosarum A34 nodC::Tn5 mutant. Phylogenetic analysis showed that Dg1 Nod proteins are positioned at the root of both α- and β-rhizobial NodABC proteins. NodA-like acyl transferases were found across the phylum Actinobacteria, but among Proteobacteria only in nodulators. Taken together, our evidence indicates an Actinobacterial origin of rhizobial Nod factors. PMID:26020781

  6. Bacterial exopolysaccharides as a modern biotechnological tool for modification of fungal laccase properties and metal ion binding.

    Science.gov (United States)

    Osińska-Jaroszuk, Monika; Jaszek, Magdalena; Starosielec, Magdalena; Sulej, Justyna; Matuszewska, Anna; Janczarek, Monika; Bancerz, Renata; Wydrych, Jerzy; Wiater, Adrian; Jarosz-Wilkołazka, Anna

    2018-03-26

    Four bacterial EPSs extracted from Rhizobium leguminosarum bv. trifolii Rt24.2, Sinorhizobium meliloti Rm1021, Bradyrhizobium japonicum USDA110, and Bradyrhizobium elkanii USDA76 were determined towards their metal ion adsorption properties and possible modification of Cerrena unicolor laccase properties. The highest magnesium and iron ion-sorption capacity (~ 42 and ~ 14.5%, respectively) was observed for EPS isolated from B. japonicum USDA110. An evident influence of EPSs on the stability of laccase compared to the control values (without EPSs) was shown after 30-day incubation at 25 °C. The residual activity of laccases was obtained in the presence of Rh76EPS and Rh1021EPS, i.e., 49.5 and 41.5% of the initial catalytic activity, respectively. This result was confirmed by native PAGE electrophoresis. The EPS effect on laccase stability at different pH (from 3.8 to 7.0) was also estimated. The most significant changes at the optimum pH value (pH 5.8) was observed in samples of laccase stabilized by Rh76EPS and Rh1021EPS. Cyclic voltamperometry was used for analysis of electrochemical parameters of laccase stabilized by bacterial EPS and immobilized on single-walled carbon nanotubes (SWCNTs) with aryl residues. Laccases with Rh76EPS and Rh1021EPS had an evident shift of the value of the redox potential compared to the control without EPS addition. In conclusion, the results obtained in this work present a new potential use of bacterial EPSs as a metal-binding component and a modulator of laccase properties especially stability of enzyme activity, which can be a very effective tool in biotechnology and industrial applications.

  7. Integrated management of foot rot of lentil using biocontrol agents under field condition.

    Science.gov (United States)

    Hannan, M A; Hasan, M M; Hossain, I; Rahman, S M E; Ismail, Alhazmi Mohammed; Oh, Deog-Hwan

    2012-07-01

    The efficacy of cowdung, Bangladesh Institute of Nuclear Agriculture (BINA)-biofertilizer, and Bangladesh Agricultural University (BAU)-biofungicide, alone or in combination, was evaluated for controlling foot rot disease of lentil. The results exhibited that BINA-biofertilizer and BAUbiofungicide (peat soil-based Rhizobium leguminosarum and black gram bran-based Trichoderma harzianum) are compatible and have combined effects in controlling the pathogenic fungi Fusarium oxysporum and Sclerotium rolfsii, which cause the root rot of lentil. Cowdung mixing with soil (at 5 t/ha) during final land preparation and seed coating with BINA-biofertilizer and BAU-biofungicide (at 2.5% of seed weight) before sowing recorded 81.50% field emergence of lentil, which showed up to 19.85% higher field emergence over the control. Post-emergence deaths of plants due to foot rot disease were significantly reduced after combined seed treatment with BINA-biofertilizer and BAU-biofungicide. Among the treatments used, only BAU-biofungicide as the seed treating agent resulted in higher plant stand (84.82%). Use of BINA-biofertilizer and BAU-biofungicide as seed treating biocontrol agents and application of cowdung in the soil as an organic source of nutrient resulted in higher shoot and root lengths, and dry shoot and root weights of lentil. BINA-biofertilizer significantly increased the number of nodules per plant and nodules weight of lentil. Seeds treating with BAUbiofungicide and BINA-biofertilizer and soil amendment with cowdung increased the biomass production of lentil up to 75.56% over the control.

  8. Rhizobium Impacts on Seed Productivity, Quality, and Protection of Pisum sativum upon Disease Stress Caused by Didymella pinodes: Phenotypic, Proteomic, and Metabolomic Traits

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    Nima Ranjbar Sistani

    2017-11-01

    Full Text Available In field peas, ascochyta blight is one of the most common fungal diseases caused by Didymella pinodes. Despite the high diversity of pea cultivars, only little resistance has been developed until to date, still leading to significant losses in grain yield. Rhizobia as plant growth promoting endosymbionts are the main partners for establishment of symbiosis with pea plants. The key role of Rhizobium as an effective nitrogen source for legumes seed quality and quantity improvement is in line with sustainable agriculture and food security programs. Besides these growth promoting effects, Rhizobium symbiosis has been shown to have a priming impact on the plants immune system that enhances resistance against environmental perturbations. This is the first integrative study that investigates the effect of Rhizobium leguminosarum bv. viceae (Rlv on phenotypic seed quality, quantity and fungal disease in pot grown pea (Pisum sativum cultivars with two different resistance levels against D. pinodes through metabolomics and proteomics analyses. In addition, the pathogen effects on seed quantity components and quality are assessed at morphological and molecular level. Rhizobium inoculation decreased disease severity by significant reduction of seed infection level. Rhizobium symbiont enhanced yield through increased seed fresh and dry weights based on better seed filling. Rhizobium inoculation also induced changes in seed proteome and metabolome involved in enhanced P. sativum resistance level against D. pinodes. Besides increased redox and cell wall adjustments light is shed on the role of late embryogenesis abundant proteins and metabolites such as the seed triterpenoid Soyasapogenol. The results of this study open new insights into the significance of symbiotic Rhizobium interactions for crop yield, health and seed quality enhancement and reveal new metabolite candidates involved in pathogen resistance.

  9. In-situ Monitoring of Plant-microbe Communication to Understand the Influence of Soil Properties on Symbiotic Biological Nitrogen Fixation

    Science.gov (United States)

    Webster, T.; Del Valle, I.; Cheng, H. Y.; Silberg, J. J.; Masiello, C. A.; Lehmann, J.

    2016-12-01

    Plant-microbe signaling is important for many symbiotic and pathogenic interactions. While this signaling often occurs in soils, very little research has evaluated the role that the soil mineral and organic matter matrix plays in plant-microbe communication. One hurdle to these studies is the lack of simple tools for evaluating how soil mineral phases and organic matter influence the availability of plant-produced flavonoids that initiate the symbiosis between nitrogen-fixing bacteria and legumes. Because of their range of hydrophobic and electrostatic properties, flavonoids represent an informative class of signaling molecules. In this presentation, we will describe studies examining the bioavailable concentrations of flavonoids in soils using traditional techniques, such as high-pressure liquid chromatography and fluorescent microbial biosensors. Additionally, we will describe our progress developing a Rhizobium leguminosarum reporter that can be deployed into soils to report on flavonoid levels. This new microbial reporter is designed so that Rhizobium only generates a volatile gas signal when it encounters a defined concentration of flavonoids. By monitoring the output of this biosensor using gas chromatography-mass spectrometry during real time during soil incubations, we are working to establish the impact of soil organic matter, pH, and mineral phases on the reception of these signaling molecules. We expect that the findings from these studies will be useful for recommending soil management strategies that can enhance the communication between legumes and nitrogen fixing bacteria. This research highlights the importance of studying the role of soil as a mediator of plant-microbe communication.

  10. Importance of new winter pea genotyp in production of the milk on family farms

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    Gordana Županac

    2009-12-01

    Full Text Available Forage pea (Pisum sativum L. is becoming more represented gorage leguminoza on the fields Republic of Croatia. Three year field trials (2003-2005 were carried out to determine the effect of seed winter pea inoculation and nitrogen top-dressing on productivity of new winter pea genotype G3 in production of milk on family farms. Just before sowing the inoculation of pea seed was performed by the variety of Rhizobium leguminosarum bv. viciae 1001 which is part of the microbiological collection of the Department of Microbiology at the Faculty of Agriculture University of Zagreb. The results of the research showed that the highest total nodule number on pea root (39.7 nodule/plant as well as nodule dry matter weight (0.203 g/plant was determined on the inoculated variant. Average highest yield of winter pea dry matter was, once more, determined on the inoculated variant (4.33 t ha-1. Total dry matter yield of winter pea and wheat mixture were ranging from 8.92 t ha-1 (control up to 10.64 t ha-1 (nitrogen top-dressing. Average highest yield of winter pea crude protein was, once more, determined on the inoculated variant (266 kg ha-1 in 2003, (672 kg ha-1 in 2004 and (853 kg ha-1 in 2005. The conclusion of this research is that the highest dry matter yield (4.33 t ha-1 and crude protein yield was obtained with the inoculation of new genotype winter pea G3.

  11. Microbial consortium role in processing liquid waste of vegetables in Keputran Market Surabaya as organic liquid fertilizer ferti-plus

    Science.gov (United States)

    Rizqi, Fauziah; Supriyanto, Agus; Lestari, Intan; Lita Indri D., L.; Elmi Irmayanti, A.; Rahmaniyah, Fadilatur

    2016-03-01

    Many activities in this market is directly proportional to increase production of vegetables waste, especially surabaya. Therefore, in this study aims to utilize liquid waste of vegetables into liquid organic fertilizer by mixing microbial consorsium. The microbial consorsium consist of Azotobacter chrococcum, Azospirillum brasilense, Rhizobium leguminosarum, Bacillus subtilis, Bacillus megaterium, Pseudomonas putida, and Pseudomonas fluorescens. Ttreatment of microbial concentrations (5%, 10%, 15%) and the length of the incubation period (7 days, 14 days, 21 days) used in this research. The parameters used are: C/N ratio, levels of CNP, and BOD value. This study uses a standard organic fertilizer value according SNI19-7030-2004, The results show the value of C/N ratio comply with the ISO standards. C levels showed an increase during the incubation period but not compare with standards. N levels that compare with standards are microbial treatment in all group concentration except control group with an incubation period of 21 days is > 7. P levels compare with the existing standards in the group of microbe concentration of 10% and 15% during the incubation period. The value of the initial BOD liquid waste of vegetable is 790.25 mg / L, this value indicates that the waste should not go into the water body. Accordingly, the results of this study can not be used as a liquid organic fertilizer, but potentially if it is used as a natural career or build natural soil. The Building natural soil is defined as the natural ingredients that can be used to improve soil properties.

  12. [The development of reagents set in the format of DNA-chip for genetic typing of strains of Vibrio cholerae].

    Science.gov (United States)

    Pudova, E A; Markelov, M L; Dedkov, V G; Tchekanova, T A; Sadjin, A I; Kirdiyashkina, N P; Bekova, M V; Deviyatkin, A A

    2014-05-01

    The necessity of development of methods of genic diagnostic of cholera is conditioned by continuation of the Seventh pandemic of cholera, taxonomic variability of strains of Vibrio cholerae involved into pandemic and also permanent danger of delivery of disease to the territory of the Russian Federation. The methods of genic diagnostic of cholera make it possible in a comparatively short time to maximally minutely characterize strains isolated from patients or their environment. The article presents information about working out reagents set for genetic typing of agents of cholera using DNA-chip. The makeup of DNA-chip included oligonucleotide probes making possible to differentiate strains of V. cholerae on serogroups and biovars and to determine their pathogenicity. The single DNA-chip makes it possible to genetically type up to 12 samples concurrently. At that, duration of analysis without accounting stage of DNA separation makes up to 5 hours. In the progress of work, 23 cholera and non-cholera strains were analyzed. The full compliance of DNA-chip typing results to previously known characteristics of strains. Hence, there is a reason to consider availability of further development of reagents set and possibility of its further application in laboratories of regional level and reference centers.

  13. Corynebacterium diphtheriae in a free-roaming red fox: case report and historical review on diphtheria in animals.

    Science.gov (United States)

    Sing, Andreas; Konrad, Regina; Meinel, Dominik M; Mauder, Norman; Schwabe, Ingo; Sting, Reinhard

    2016-08-01

    Corynebacterium diphtheriae, the classical causative agent of diphtheria, is considered to be nearly restricted to humans. Here we report the first finding of a non-toxigenic C. diphtheriae biovar belfanti strain in a free-roaming wild animal. The strain obtained from the subcutis and mammary gland of a dead red fox (Vulpes vulpes) was characterized by biochemical and molecular methods including MALDI-TOF and Multi Locus Sequence Typing. Since C. diphtheriae infections of animals, usually with close contact to humans, are reported only very rarely, an intense review comprising also scientific literature from the beginning of the 20th century was performed. Besides the present case, only 11 previously reported C. diphtheriae animal infections could be verified using current scientific criteria. Our report is the first on the isolation of C. diphtheriae from a wildlife animal without any previous human contact. In contrast, the very few unambiguous publications on C. diphtheriae in animals referred to livestock or pet animals with close human contact. C. diphtheriae carriage in animals has to be considered as an exceptionally rare event.

  14. [ON THE ORIGIN OF HYPERVIRULENCE OF THE CAUSATIVE AGENT OF PLAGUE].

    Science.gov (United States)

    Anisimov, N V; Kislichkina, A A; Platonov, M E; Evseeva, V V; Kadnikova, L A; Lipatnikova, N A; Bogun, A G; Dentovskaya, S V; Anisimov, A P

    2016-01-01

    The attempt to combine Yersinia pseudotuberculosis and Yersinia pestis into one species has been unsupported by microbiologists due to the specific features of the epidemiology and clinical presentations of their induced diseases and to basic differences in their virulence. Pseudotuberculosis is predominantly a relatively mild human intestinal infection transmitted through contaminated food and plague is an acute generalized disease with high mortality, which is most frequently transmitted by the bites of infected fleas. Y. pestis hypervirulence, the ability of single bacteria to ensure the development of predagonal bacteriemia in rodents, which is sufficient to contaminate the fleas, is one of the main events during pathogen adaptation to a new ecological niche. By analyzing the data of molecular typing of the representative kits of naturally occurring Y. pestis isolates, the authois consider the issues of formation of intraspecies groups with universal hypervirulence, as well as biovars that are highly virulent only to their major host. A strategy for searching for selective virulence factors, the potential molecular targets for vaccination and etiotropic treatment of plague, is discussed.

  15. [Mexican phenotype and genotype Vibrio cholerae 01].

    Science.gov (United States)

    Giono, S; Gutiérrez Cogno, L; Rodríguez Angeles, G; del Rio Zolezzi, A; Valdespino González, J L; Sepúlveda Amor, J

    1995-01-01

    This paper presents the phenotypical and genotypical characterization of 26922 Vibrio cholerae 01 strains isolated in Mexico from 1991 to 1993. All strains isolated were El Tor biovar. Strains were sensitive to antibiotics excluding furazolidone, streptomycin and sulfisoxasole to which we found resistance in 97% and we are using this characteristic as epidemiological markers. We detected a marked change in frequency of Inaba serotype from 1991, when it was dominant, with 99.5%, until 1992 when Ogawa serotype turned to be dominant with 95% of isolates. All Vibrio cholerae 01 strains, except one Ogawa strain, were to igenic, and V. choleraeno 01 were not toxigenic by ELISA, PCR and cell culture tests. Dominant ribotype was 5, but we found some strains with 6a pattern and two with ribotype 12. We are searching for ribotype 2 among hemolytic strains in order to learn if there is any relation to Gulf Coast strains prevalent in the USA, but until now we have not found any V. cholerae ribotype 2 in our isolates. Even if rapid tests are recommended for immediate diagnosis of cholera, it is necessary to continue bacterial isolation in order to have strains for phenotyping and genotyping studies that may support epidemiological analysis.

  16. Expedient synthesis of C-aryl carbohydrates by consecutive biocatalytic benzoin and aldol reactions.

    Science.gov (United States)

    Hernández, Karel; Parella, Teodor; Joglar, Jesús; Bujons, Jordi; Pohl, Martina; Clapés, Pere

    2015-02-16

    The introduction of aromatic residues connected by a C-C bond into the non-reducing end of carbohydrates is highly significant for the development of innovative structures with improved binding affinity and selectivity (e.g., C-aril-sLex). In this work, an expedient asymmetric "de novo" synthetic route to new aryl carbohydrate derivatives based on two sequential stereoselectively biocatalytic carboligation reactions is presented. First, the benzoin reaction of aromatic aldehydes to dimethoxyacetaldehyde is conducted, catalyzed by benzaldehyde lyase from Pseudomonas fluorescens biovar I. Then, the α-hydroxyketones formed are reduced by using NaBH4 yielding the anti diol. After acetal hydrolysis, the aldol addition of dihydroxyacetone, hydroxyacetone, or glycolaldehyde catalyzed by the stereocomplementary D-fructose-6-phosphate aldolase and L-rhamnulose-1-phosphate aldolase is performed. Both aldolases accept unphosphorylated donor substrates, avoiding the need of handling the phosphate group that the dihydroxyacetone phosphate-dependent aldolases require. In this way, 6-C-aryl-L-sorbose, 6-C-aryl-L-fructose, 6-C-aryl-L-tagatose, and 5-C-aryl-L-xylose derivatives are prepared by using this methodology. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Rapid and specific identification of Brucella abortus using the loop-mediated isothermal amplification (LAMP) assay.

    Science.gov (United States)

    Kang, Sung-Il; Her, Moon; Kim, Ji-Yeon; Lee, Jin Ju; Lee, Kichan; Sung, So-Ra; Jung, Suk Chan

    2015-06-01

    A rapid and accurate diagnosis of brucellosis is required to reduce and prevent the spread of disease among animals and the risk of transfer to humans. In this study, a Brucella abortus-specific (Ba) LAMP assay was developed, that had six primers designed from the BruAb2_0168 region of chromosome I. The specificity of this LAMP assay was confirmed with Brucella reference strains, B. abortus vaccine strains, B. abortus isolates and phylogenetically or serologically related strains. The detection limit of target DNA was up to 20 fg/μl within 60 min. The sensitivity of the new LAMP assay was equal to or slightly higher than other PCR based assays. Moreover, this Ba-LAMP assay could specifically amplify all B. abortus biovars compared to previous PCR assays. To our knowledge, this is the first report of specific detection of B. abortus using a LAMP assay. The Ba-LAMP assay can offer a rapid, sensitive and accurate diagnosis of bovine brucellosis in the field. Copyright © 2015 Elsevier Ltd. All rights reserved.

  18. Brucella abortusΔcydCΔcydD and ΔcydCΔpurD double-mutants are highly attenuated and confer long-term protective immunity against virulent Brucella abortus.

    Science.gov (United States)

    Truong, Quang Lam; Cho, Youngjae; Park, Soyeon; Kim, Kiju; Hahn, Tae-Wook

    2016-01-04

    We constructed double deletion (ΔcydCΔcydD and ΔcydCΔpurD) mutants from virulent Brucella abortus biovar 1 field isolate (BA15) by deleting the genes encoding an ATP-binding cassette-type transporter (cydC and cydD genes) and a phosphoribosylamine-glycine ligase (purD). Both BA15ΔcydCΔcydD and BA15ΔcydCΔpurD double-mutants exhibited significant attenuation of virulence when assayed in murine macrophages or in BALB/c mice. Both double-mutants were readily cleared from spleens by 4 weeks post-inoculation even when inoculated at the dose of 10(8) CFU per mouse. Moreover, the inoculated mice showed no splenomegaly, which indicates that the mutants are highly attenuated. Importantly, the attenuation of in vitro and in vivo growth did not impair the ability of these mutants to confer long-term protective immunity in mice against challenge with B. abortus strain 2308. Vaccination of mice with either mutant induced humoral and cell-mediated immune responses, and provided significantly better protection than commercial B. abortus strain RB51 vaccine. These results suggest that highly attenuated BA15ΔcydCΔcydD and BA15ΔcydCΔpurD mutants can be used effectively as potential live vaccine candidates against bovine brucellosis. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

  19. Investigations on brucellosis in Egyptian Baladi Does with emphasis on evaluation of diagnostic techniques.

    Science.gov (United States)

    el-Razik, K A Abd; Desouky, H M; Ahmed, W M

    2007-01-15

    Investigations were carried out on caprine brucellosis in a costal area in Egypt. A total number of 577 Baladi Does was examined for Brucella infection using different serological tests. Specimens were taken from seropositive obligatory slaughtered Does (No = 33) for Brucella isolation, histopathological examination, Polymerase Chain Reaction (PCR) assay and determination of serum copper (Cu), zinc(Zn) and iron (Fe) concentrations. Results indicated that the incidence of brucellosis was 3.0-5.0%, by using the different serological tests. Buffered Acidified Plate Antigen Test (BAPAT) is of the highest sensitivity followed by Rose Bengal Plate Test (RBPT), L-ELISA, Complement Fixation Test (CFT), P-ELISA, Rivanol test (RVT) and Tube Agglutination Test (TAT). In seropostive Does, Brucella melitensis biovar 3 was isolated from 78.78% and PCR yielded expected products in 81.81%. Moreover, granulomatous endometritis, lymphocytic mastitis and lymphoid depletion in both lymph nodes and spleen were evident together with significant (p<0.001) decreases in serum Cu, Zn and Fe concentrations. In conclusion, more attention should be paid to goat in brucellosis epidemiology in the application of national program of brucella control and eradication.

  20. Strain-related acid production by oral streptococci

    DEFF Research Database (Denmark)

    de Soet, JJ; Nyvad, Bente; Kilian, Mogens

    2000-01-01

    Acid production, in particular at low pH, is thought to be an important ecological determinant in dental caries. The aim of the present study was to determine the acid producing capability at different pH levels of 47 streptococcal strains, representing 9 species, isolated from human dental plaque....... The bacteria were grown until mid log-phase under anaerobic conditions and acid production was measured in a pH-stat system at pH 7.0, 6.0, 5.5 and 5.0. At all pH values, the mean velocity of acid production (V(ap)) by Streptococcus mutans and S. sobrinus was significantly higher (p... that of the other oral streptococci, including S. mitis, S. oralis, S. gordonii, S. sanguis, S. intermedius, S. anginosus, S. constellatus, and S. vestibularis. However, the V(ap) of some strains of S. mitis biovar 1 and S. oralis, particularly at pH values of 7.0 and 6.0, exceeded that of some strains of S. mutans...

  1. Validation of the multiplex PCR for identification of Brucella spp.

    Directory of Open Access Journals (Sweden)

    Lívia de Lima Orzil

    2016-05-01

    Full Text Available ABSTRACT: A multiplex PCR technique for detection of Brucella spp. in samples of bacterial suspension was validated as a complementary tool in the diagnosis of the disease. This technique allows the characterization of the agent without performing biochemical tests, which greatly reduces the time for a final diagnosis, and provides more security for the analyst by reducing the time of exposure to microorganisms. The validation was performed in accordance with the Manual of Diagnostic Tests from OIE (2008 and following the requirements present in the ABNT NBR ISO/IEC 17025:2005. The mPCR validated in this study identified the different species of Brucella ( Brucella abortus , B. suis , B. ovis e B. melitensis of bacterial suspension obtained from the slaughterhouse samples, as well as distinguished the biovars (1, 2 e 4; 3b, 5, 6 e 9 of B. abortus in grouped form and differentiated the field strains from vaccine strains, as a quick, useful and less expensive technique in diagnosis of brucellosis in Brazil.

  2. Analysis of temperature-dependent changes in the metabolism of Yersinia pestis.

    Science.gov (United States)

    Navid, Ali; Almaas, Eivind

    2008-03-01

    The gram-negative bacterium Yersinia pestis is the aetiological agent of bubonic plague, a zoonotic infection that occurs through the bite of a flea. It has long been known that Y. pestis has different metabolic needs upon transition from the flea gut environment (26 C) to that of a mammalian host (37 C). To study this and other outstanding questions about metabolic function of Y. pestis, we used the available genomic, biochemical and physiological data to develop a constraint-based flux balance model of metabolism in the avirulent 91001 strain (biovar Mediaevalis) of this organism. Utilizing two sets of whole-genome DNA microarray expression data, we examined the system level changes that occur when Y. pestis acclimatizes to temperature shifts. Our results point to fundamental changes in its oxidative metabolism of sugars and use of amino acids, in particular that of arginine. This behavior is indicative of an inefficient metabolism that could be caused by adaptation to life in a nutrient rich environment.

  3. Typing methods for the plague pathogen, Yersinia pestis.

    Science.gov (United States)

    Lindler, Luther E

    2009-01-01

    Phenotypic and genotypic methodologies have been used to differentiate the etiological agent of plague, Yersinia pestis. Historically, phenotypic methods were used to place isolates into one of three biovars based on nitrate reduction and glycerol fermentation. Classification of Y. pestis into genetic subtypes is problematic due to the relative monomorphic nature of the pathogen. Resolution into groups is dependent on the number and types of loci used in the analysis. The last 5-10 years of research and analysis in the field of Y. pestis genotyping have resulted in a recognition by Western scientists that two basic types of Y. pestis exist. One type, considered to be classic strains that are able to cause human plague transmitted by the normal flea vector, is termed epidemic strains. The other type does not typically cause human infections by normal routes of infection, but is virulent for rodents and is termed endemic strains. Previous classification schemes used outside the Western hemisphere referred to these latter strains as Pestoides varieties of Y. pestis. Recent molecular analysis has definitely shown that both endemic and epidemic strains arose independently from a common Yersinia pseudotuberculosis ancestor. Currently, 11 major groups of Y. pestis are defined globally.

  4. Targeted enrichment of ancient pathogens yielding the pPCP1 plasmid of Yersinia pestis from victims of the Black Death.

    Science.gov (United States)

    Schuenemann, Verena J; Bos, Kirsten; DeWitte, Sharon; Schmedes, Sarah; Jamieson, Joslyn; Mittnik, Alissa; Forrest, Stephen; Coombes, Brian K; Wood, James W; Earn, David J D; White, William; Krause, Johannes; Poinar, Hendrik N

    2011-09-20

    Although investigations of medieval plague victims have identified Yersinia pestis as the putative etiologic agent of the pandemic, methodological limitations have prevented large-scale genomic investigations to evaluate changes in the pathogen's virulence over time. We screened over 100 skeletal remains from Black Death victims of the East Smithfield mass burial site (1348-1350, London, England). Recent methods of DNA enrichment coupled with high-throughput DNA sequencing subsequently permitted reconstruction of ten full human mitochondrial genomes (16 kb each) and the full pPCP1 (9.6 kb) virulence-associated plasmid at high coverage. Comparisons of molecular damage profiles between endogenous human and Y. pestis DNA confirmed its authenticity as an ancient pathogen, thus representing the longest contiguous genomic sequence for an ancient pathogen to date. Comparison of our reconstructed plasmid against modern Y. pestis shows identity with several isolates matching the Medievalis biovar; however, our chromosomal sequences indicate the victims were infected with a Y. pestis variant that has not been previously reported. Our data reveal that the Black Death in medieval Europe was caused by a variant of Y. pestis that may no longer exist, and genetic data carried on its pPCP1 plasmid were not responsible for the purported epidemiological differences between ancient and modern forms of Y. pestis infections.

  5. Populations of predators and parasitoids of Bemisia tabaci (Hemiptera: Aleyrodidae) after the application of eight biorational insecticides in vegetable crops.

    Science.gov (United States)

    Simmons, Alvin M; Shaaban, Abd-Rabou

    2011-08-01

    The sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), is an important pest of vegetables and many other crops worldwide. Eight biorational insecticides (based on oil, plant derivatives, insect growth regulator and fungus) were evaluated in the field for their influence on populations of six natural enemies of B. tabaci. Natural populations of two predators [Chrysoperla carnea Stephen (Neuroptera: Chrysopidae) and Orius spp. (Hemiptera: Anthocoridae)] and two genera of parasitoids [Encarsia spp. and Eretmocerus spp. (Hymenoptera: Aphelinidae)] were evaluated in eggplant (Solanum melongena L.). Also, augmented field populations of three predators [C. carnea, Coccinella undecimpunctata L. (Coleoptera: Coccinellidae) and Macrolophus caliginosus (Wagner) (Hemiptera: Miridae)] were evaluated in cabbage (Brassica oleracea var. capitata L.), cucumber (Cucumis sativus L.) and squash (Cucurbita pepo L.). Regardless of natural enemy or crop, jojoba oil, Biovar and Neemix had the least effect on abundance of the natural enemies in comparison with the other insecticides during a 14 day evaluation period. Conversely, Admiral, KZ oil, Mesrona oil, Mesrona oil + sulfur and natural oil had a high detrimental effect on abundance of the natural enemies. These results demonstrate the differential effects of biorational insecticides for whitefly control on predators and parasitoids in the field. This article is a US Government work and is in the public domain in the USA. Published 2011 by John Wiley & Sons, Ltd.

  6. An overview on avian influenza

    Directory of Open Access Journals (Sweden)

    Nelson Rodrigo da Silva Martins

    2012-06-01

    Full Text Available Avian influenza (AI is considered an exotic disease in the Brazilian poultry industry, according to the National Avian Health Program (PNSA, with permanent monitoring of domestic, exotic and native avian species. Brazil presents privileged environmental conditions of reduced risk. In addition, all commercial poultry and conservation holdings are registered in state or national inventories and geographically located (GPS for health control. Poultry health standards are adopted for the conformity to the international market, mostly for the intensified poultry destined for exportation, but also for companion exotic and native conservation facilities. Guidelines for monitoring and the diagnosis of AI are published by the PNSA and follow the standards proposed by the international health code (World Organization for Animal Health, Organization International des Epizooties - OIE and insure the free of status for avian influenza virus (AIV of LPAIV-low pathogenicity AIV and HPAIV-high pathogenicity AIV. In addition, the infections by mesogenic and velogenic Newcastle disease virus, Mycoplasma gallisepticum, M. synoviae and M. meleagridis, Salmonella enteric subspecies enterica serovar Gallinarum biovars Gallinarum and Pullorum are eradicated from reproduction. Controlled infections by S.enterica subspecies enterica serovars Enteritidis and Typhimurium are monitored for breeders. The vaccination of chickens in ovo or at hatch against Marek's disease is mandatory. Broiler production is an indoor activity, confinement which insures biosecurity, with safe distances from the potential AIV reservoir avian species. Worldwide HPAIV H5N1 notifications to the OIE, in March 2011, included 51 countries.

  7. A vegan or vegetarian diet substantially alters the human colonic faecal microbiota.

    Science.gov (United States)

    Zimmer, J; Lange, B; Frick, J-S; Sauer, H; Zimmermann, K; Schwiertz, A; Rusch, K; Klosterhalfen, S; Enck, P

    2012-01-01

    Consisting of ≈10(14) microbial cells, the intestinal microbiota represents the largest and the most complex microbial community inhabiting the human body. However, the influence of regular diets on the microbiota is widely unknown. We examined faecal samples of vegetarians (n=144), vegans (n=105) and an equal number of control subjects consuming ordinary omnivorous diet who were matched for age and gender. We used classical bacteriological isolation, identification and enumeration of the main anaerobic and aerobic bacterial genera and computed absolute and relative numbers that were compared between groups. Total counts of Bacteroides spp., Bifidobacterium spp., Escherichia coli and Enterobacteriaceae spp. were significantly lower (P=0.001, P=0.002, P=0.006 and P=0.008, respectively) in vegan samples than in controls, whereas others (E. coli biovars, Klebsiella spp., Enterobacter spp., other Enterobacteriaceae, Enterococcus spp., Lactobacillus spp., Citrobacter spp. and Clostridium spp.) were not. Subjects on a vegetarian diet ranked between vegans and controls. The total microbial count did not differ between the groups. In addition, subjects on a vegan or vegetarian diet showed significantly (P=0.0001) lower stool pH than did controls, and stool pH and counts of E. coli and Enterobacteriaceae were significantly correlated across all subgroups. Maintaining a strict vegan or vegetarian diet results in a significant shift in the microbiota while total cell numbers remain unaltered.

  8. Thermostable cross-protective subunit vaccine against Brucella species.

    Science.gov (United States)

    Cherwonogrodzky, John W; Barabé, Nicole D; Grigat, Michelle L; Lee, William E; Poirier, Robert T; Jager, Scott J; Berger, Bradley J

    2014-12-01

    A subunit vaccine candidate was produced from Brucella suis 145 (biovar 4; expressing both the A antigen of Brucella abortus and the M antigen of Brucella melitensis). The preparation consisted mostly of polysaccharide (PS; >90% [wt/wt]; both cell-associated PS and exo-PS were combined) and a small amount of protein (1 to 3%) with no apparent nucleic acids. Vaccinated mice were protected (these had a statistically significant reduction in bacterial colonization compared to that of unvaccinated controls) when challenged with representative strains of three Brucella species most pathogenic for humans, i.e., B. abortus, B. melitensis, and B. suis. As little as 1 ng of the vaccine, without added adjuvant, protected mice against B. suis 145 infection (5 × 10(5) CFU), and a single injection of 1 μg of this subunit vaccine protected mice from B. suis 145 challenge for at least 14 months. A single immunization induced a serum IgG response to Brucella antigens that remained elevated for up to 9 weeks. The use of heat (i.e., boiling-water bath, autoclaving) in the vaccine preparation showed that it was thermostable. This method also ensured safety and security. The vaccine produced was immunogenic and highly protective against multiple strains of Brucella and represents a promising candidate for further evaluation. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  9. Molecular epidemiological investigation of Brucella melitensis circulating in Mongolia by MLVA16.

    Science.gov (United States)

    Kang, Sung-Il; Her, Moon; Erdenebaataar, Janchivdorj; Vanaabaatar, Batbaatar; Cho, Hyorim; Sung, So-Ra; Lee, Jin Ju; Jung, Suk Chan; Park, Yong Ho; Kim, Ji-Yeon

    2017-02-01

    Mongolia has a high incidence of brucellosis in human and animals due to livestock husbandry. To investigate the genetic characteristics of Mongolian B. melitensis, an MLVA (multi-locus variable-number tandem-repeat analysis)-16 assay was performed with 94 B. melitensis isolates. They were identified as B. melitensis biovar (bv.) 1 (67), 3 (10) and Rev. 1 vaccine strains (17) using a classical biotyping and multiplex PCR. In genotyping, three human isolates were grouped at 2 genotypes with sheep isolates, and it implies that B. melitensis are cross-infected between human and livestock. In the parsimony analysis, Mongolian B. melitensis isolates had high genetic similarity with Chinese strains, likely due to the geographical proximity, clustered distinctively as compared with other foreign isolates. B. melitensis Rev. 1 vaccine strains were divided into 4 genotypes with 92% similarity. In the analysis of Rev.1 strains, the risk of mutation of vaccine strain might not be overlooked. Animal quarantines should be strengthened to prevent the spread of Brucella species among adjacent countries. Copyright © 2016 Elsevier Ltd. All rights reserved.

  10. Molecular strain typing of Brucella abortus isolates from Italy by two VNTR allele sizing technologies.

    Science.gov (United States)

    De Santis, Riccardo; Ancora, Massimo; De Massis, Fabrizio; Ciammaruconi, Andrea; Zilli, Katiuscia; Di Giannatale, Elisabetta; Pittiglio, Valentina; Fillo, Silvia; Lista, Florigio

    2013-10-01

    Brucellosis, one of the most important re-emerging zoonoses in many countries, is caused by bacteria belonging to the genus Brucella. Furthermore these bacteria represent potential biological warfare agents and the identification of species and biovars of field strains may be crucial for tracing back source of infection, allowing to discriminate naturally occurring outbreaks instead of bioterrorist events. In the last years, multiple-locus variable-number tandem repeat analysis (MLVA) has been proposed as complement of the classical biotyping methods and it has been applied for genotyping large collections of Brucella spp. At present, the MLVA band profiles may be resolved by automated or manual procedures. The Lab on a chip technology represents a valid alternative to standard genotyping techniques (as agarose gel electrophoresis) and it has been previously used for Brucella genotyping. Recently, a new high-throughput genotyping analysis system based on capillary gel electrophoresis, the QIAxcel, has been described. The aim of the study was to evaluate the ability of two DNA sizing equipments, the QIAxcel System and the Lab chip GX, to correctly call alleles at the sixteen loci including one frequently used MLVA assay for Brucella genotyping. The results confirmed that these technologies represent a meaningful advancement in high-throughput Brucella genotyping. Considering the accuracy required to confidently resolve loci discrimination, QIAxcel shows a better ability to measure VNTR allele sizes compared to LabChip GX.

  11. A serological survey for brucellosis in reindeer in Finnmark county, northern Norway

    Directory of Open Access Journals (Sweden)

    Kjetil Åsbakk

    1999-04-01

    Full Text Available During September-December, 1990 to 1994, serum samples from a total of 5792 semi-domesticated reindeer (Rangifer tarandm tarandm from Finnmark county, northern Norway, were screened for brucellosis on an indirect ELISA. There were no serologically positive animals. Twenty six of the animals had levels of antibodies detectable on the ELISA and were classed as suspicious, but the ELISA optical density readings were low compared to the readings for reindeer that were both culture positive and seropositive for Brucella suis biovar 4. When assayed on the standard tube agglutination test (STAT, all the 26 animals were seronegative. When absorbed with cells of Yersinia enterocolitica 0-9, the antibody detectable on the ELISA could be removed to a great extent from most of the sera, indicating previous or ongoing exposure to bacteria serologically cross-reacting with Brucella in these animals. We concluded that brucellosis was not present among reindeer in Finnmark during this study. This is supported by the absence of any reports of brucellosis among reindeer in Norway.

  12. Different region analysis for genotyping Yersinia pestis isolates from China.

    Directory of Open Access Journals (Sweden)

    Yanjun Li

    Full Text Available BACKGROUND: DFR (different region analysis has been developed for typing Yesinia pestis in our previous study, and in this study, we extended this method by using 23 DFRs to investigate 909 Chinese Y. pestis strains for validating DFR-based genotyping method and better understanding adaptive microevolution of Y. pestis. METHODOLOGY/PRINCIPAL FINDINGS: On the basis of PCR and Bionumerics data analysis, 909 Y. pestis strains were genotyped into 32 genomovars according to their DFR profiles. New terms, Major genomovar and Minor genomovar, were coined for illustrating evolutionary relationship between Y. pestis strains from different plague foci and different hosts. In silico DFR profiling of the completed or draft genomes shed lights on the evolutionary scenario of Y. pestis from Y. pseudotuberculosis. Notably, several sequenced Y. pestis strains share the same DFR profiles with Chinese strains, providing data for revealing the global plague foci expansion. CONCLUSIONS/SIGNIFICANCE: Distribution of Y. pestis genomovars is plague focus-specific. Microevolution of biovar Orientalis was deduced according to DFR profiles. DFR analysis turns to be an efficient and inexpensive method to portrait the genome plasticity of Y. pestis based on horizontal gene transfer (HGT. DFR analysis can also be used as a tool in comparative and evolutionary genomic research for other bacteria with similar genome plasticity.

  13. Molecular diversity of leuconostoc mesenteroides and leuconostoc citreum isolated from traditional french cheeses as revealed by RAPD fingerprinting, 16S rDNA sequencing and 16S rDNA fragment amplification.

    Science.gov (United States)

    Cibik, R; Lepage, E; Talliez, P

    2000-06-01

    For a long time, the identification of the Leuconostoc species has been limited by a lack of accurate biochemical and physiological tests. Here, we use a combination of RAPD, 16S rDNA sequencing, and 16S rDNA fragment amplification with specific primers to classify different leuconostocs at the species and strain level. We analysed the molecular diversity of a collection of 221 strains mainly isolated from traditional French cheeses. The majority of the strains were classified as Leuconostoc mesenteroides (83.7%) or Leuconostoc citreum (14%) using molecular techniques. Despite their presence in French cheeses, the role of L. citreum in traditional technologies has not been determined, probably because of the lack of strain identification criteria. Only one strain of Leuconostoc lactis and Leuconostoc fallax were identified in this collection, and no Weissella paramesenteroides strain was found. However, dextran negative variants of L. mesenteroides, phenotypically misclassified as W. paramesenteroides, were present. The molecular techniques used did not allow us to separate strains of the three L. mesenteroides subspecies (mesenteroides, dextranicum and cremoris). In accordance with previously published results, our findings suggest that these subspecies may be classified as biovars. Correlation found between phenotypes dextranicum and mesenteroides of L. mesenteroides and cheese technology characteristics suggests that certain strains may be better adapted to particular technological environments.

  14. First report of Corynebacterium pseudotuberculosis from caseous lymphadenitis lesions in Black Alentejano pig (Sus scrofa domesticus).

    Science.gov (United States)

    Oliveira, Manuela; Barroco, Cynthia; Mottola, Carla; Santos, Raquel; Lemsaddek, Abdelhak; Tavares, Luis; Semedo-Lemsaddek, Teresa

    2014-09-21

    Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis, a common disease in small ruminant populations throughout the world and responsible for a significant economic impact for producers. To our knowledge, this is the first characterization of C. pseudotuberculosis from caseous lymphadenitis lesions in Black Alentejano pig (Sus scrofa domesticus). In this study, phenotypic and genotypic identification methods allocated the swine isolates in C. pseudotuberculosis biovar ovis. The vast majority of the isolates were able to produce phospholipase D and were susceptible to most of the antimicrobial compounds tested. Macrorestriction patterns obtained by Pulsed Field Gel Electrophoresis (PFGE) grouped the C. pseudotuberculosis in two clusters with a high similarity index, which reveals their clonal relatedness. Furthermore, swine isolates were compared with C. pseudotuberculosis from caprines and PFGE patterns also showed high similarity, suggesting the prevalence of dominant clones and a potential cross-dissemination between these two animal hosts. This work represents the first report of Corynebacterium pseudotuberculosis from caseous lymphadenitis lesions in Black Alentejano pig and alerts for the importance of the establishment of suitable control and sanitary management practices to control the infection and avoid further dissemination of this important pathogen to other animal hosts.

  15. Identifikasi Brucella abortus Isolat Lokal dengan Brucella abortus Strain Specific-Polymerase Chain Reaction (IDENTIFICATION OF LOCAL ISOLATES OF BRUCELLA ABORTUS USING BRUCELLA ABORTUS STRAIN SPECIFIC-POLYMERASE CHAIN REACTION ASSAY

    Directory of Open Access Journals (Sweden)

    Susan Maphilindawati Noor

    2014-10-01

    Full Text Available Brucella abortus Strain Specific-Polymerase Chain Reaction (BaSS-PCR is a single multiplex PCRtechnique which able to identify and differentiate between Brucella abortus field strains (biovar 1, 2, and4, B. abortus vaccine strains, Brucella species, and non-Brucella species. In this study, BaSS-PCR wasapplied to identify local isolates of B. abortus in order to investigate the B. abortus strains that infectedcattle in Indonesia. Fifty local strains of B.abortus isolated from infected cattle in Java (Jakarta andBandung, South Sulawesi (Maros, East Nusa Tenggara (Kupang and Belu were used in this study. TheDNA bands were observed by agarose gel in the presence of ethidium bromide. Identification was performedbased on the size and number of DNA products amplified by PCR from each isolates. The results showedthat the 50 isolates were of B. abortus field strains. This finding showed that the cause of bovine brucellosisin Indonesia is B. abortus field strains.

  16. 16S rRNA and Omp31 Gene Based Molecular Characterization of Field Strains of B. melitensis from Aborted Foetus of Goats in India

    Science.gov (United States)

    Singh, Ajay; Gupta, Vivek Kumar; Kumar, Amit; Singh, Vikas Kumar; Nayakwadi, Shivasharanappa

    2013-01-01

    Brucellosis is a reemerging infectious zoonotic disease of worldwide importance. In human, it is mainly caused by Brucella melitensis, a natural pathogen for goats. In India, a large number of goats are reared in semi-intensive to intensive system within the close vicinity of human being. At present, there is no vaccination and control strategy for caprine brucellosis in the country. Thus, to formulate an effective control strategy, the status of etiological agent is essential. To cope up with these, the present study was conducted to isolate and identify the prevalent Brucella species in caprine brucellosis in India. The 30 samples (fetal membrane, fetal stomach content and vaginal swabs) collected throughout India from the aborted fetus of goats revealed the isolation of 05 isolates all belonging to Brucella melitensis biovars 3. All the isolates produced amplification products of 1412 and 720 bp in polymerase chain reaction with genus and species specific 16S rRNA and omp31 gene based primers, respectively. Moreover, the amplification of omp31 gene in all the isolates confirmed the presence of immuno dominant outer membrane protein (31 kDa omp) in all the field isolates of B. melitensis in aborted foetus of goats in India. These findings can support the development of omp31 based specific serodiagnostic test as well as vaccine for the control of caprine brucellosis in India. PMID:24453799

  17. Investigation of the bacteriophage community in induced lysates of undefined mesophilic mixed-strain DL-cultures using classical and metagenomic approaches.

    Science.gov (United States)

    Muhammed, Musemma K; Olsen, Mette L; Kot, Witold; Neve, Horst; Castro-Mejía, Josué L; Janzen, Thomas; Hansen, Lars H; Nielsen, Dennis S; Sørensen, Søren J; Heller, Knut J; Vogensen, Finn K

    2018-05-02

    To investigate the notion that starter cultures can be a reservoir of bacteriophages (phages) in the dairy environment, strains of three DL-starters (undefined mesophilic mixed-strain starters containing Lactococcus lactis subsp. lactis biovar. diacetylactis and Leuconostoc species) were selected and induced by mitomycin C, and the whole starters were induced spontaneously as well as by mitomycin C. Frequency of induction of 17%, 26% and 12% was estimated among the isolates of the three starters, with majority of the induced phages mostly showing morphological similarity to known P335 phages, and with a fraction of them showing atypical features. Sequences of P335 quasi-species phages were found to be the most frequent entities in almost all metaviromes derived from the induced lysates. However, sequences of Sk1virus phages (previously 936 phages) were emerged as the predominant entities following spontaneous induction of one of the starters, suggesting a phage-carrier state. Sequences of other phages such as 949, 1706, C2virus (previously c2 phages) and Leuconostoc species could also be observed but with a lower relative frequency. Taken together, the majority of the P335 quasi-species phages could represent the induced viral community of the starters and the remaining phage groups mainly represent the background ambient viral community. Copyright © 2018 Elsevier B.V. All rights reserved.

  18. Human brucellosis in France in the 21st century: Results from national surveillance 2004-2013.

    Science.gov (United States)

    Mailles, A; Garin-Bastuji, B; Lavigne, J P; Jay, M; Sotto, A; Maurin, M; Pelloux, I; O'Callaghan, D; Mick, V; Vaillant, V; De Valk, H

    2016-12-01

    Brucellosis is a bacterial zoonotic disease mainly transmitted to humans by ruminants. In France, brucellosis has disappeared from ruminants herds. Human brucellosis surveillance is performed through mandatory notification and the national reference center. We report the results of human brucellosis surveillance from 2004 to 2013 with regards to epidemiological, clinical and microbiological data. A total of 250 cases were notified, making an annual incidence of 0.3 cases per million inhabitants. Brucella melitensis biovar 3 was the most frequently identified bacterium (79% of isolated strains). In total, 213 (85%) cases had been contaminated abroad in endemic countries. In 2012, an episode of re-emergence of brucellosis in cattle occurred in Haute-Savoie, in the French Alps, and was responsible for 2 human cases. Brucellosis has become a disease of travelers in France. However, maintaining a stringent epidemiological surveillance is necessary to be able to early detect any local re-emergence in humans or animals. The multidisciplinary surveillance was implemented in France years ago and is a successful example of the One Health Concept. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  19. Brucellosis Outbreak in Children and Adults in Two Areas in Israel

    Science.gov (United States)

    Megged, Orli; Chazan, Bibiana; Ganem, Atef; Ayoub, Abeer; Yanovskay, Anna; Sakran, Waheeb; Miron, Dan; Dror-Cohen, Ahuva; Kennes, Yoram; Berdenstein, Svetlana; Glikman, Daniel

    2016-01-01

    Two parallel outbreaks of Brucella melitensis infection occurred in 2014 in two geographical areas in Israel. In two medical centers in northern Israel and one medical center in Jerusalem, 102 patients (58 children, 47 adults) were diagnosed with brucellosis. Most patients (N = 76, 72%) were Muslim Arabs, 28 (27%) were Druze, and one was Jewish. The source of infection was often traced to cheese from the Palestinian Authority. Biovar-1 was evident in 98% in northern Israel but only in 42% in Jerusalem. Most common manifestations were fever (82%) and osteoarticular symptoms (49%). The major differences between the geographic areas were ethnicity and duration until diagnosis. Compared with adults, children had higher rates of hospitalization (93% versus 64%, P = 0.001), osteoarticular symptoms (60% versus 36%, P = 0.05), elevated alanine aminotransferase (12% versus 0%, P = 0.01), and lower C-reactive protein (2.28 ± 2.08 versus 5.57 ± 6.3l mg/dL, P = 0.001). Two unrelated brucellosis outbreaks occurred in 2014 in two different geographic areas of Israel and were limited to sections of the Arab and Druze populations. Most of the demographic and clinical aspects of patients were not affected by geographic variability. Clinical and laboratory differences were found between children and adults emphasizing the nonuniformity of the disease in different age groups. Effective control of unpasteurized dairy foods, health education programs, and improved regional cooperation are required to control brucellosis in Israel. PMID:27114301

  20. The formation of microorganism communities in the soil under the effect of chitosan and runner bean (Phaseolus coccineus L. cultivation

    Directory of Open Access Journals (Sweden)

    Danuta Pięta

    2013-12-01

    .sp. phaseoli- bean pathogen were found. The healthiness of plants grown in soil treated with chitosan was significantly better as compared to the control. The populations of antagonistic microorganisms formed in the soil in these treatments probably limited the growth of pathogenic fungus.

  1. Efeito da solarização sobre propriedades físicas, químicas e biológicas de solos

    Directory of Open Access Journals (Sweden)

    R. Ghini

    2003-02-01

    Full Text Available A solarização é um método de desinfestação que consiste na cobertura do solo com um filme de polietileno transparente, durante o período de intensa radiação solar, e atua por meio do aumento da temperatura do solo. Quatro ensaios foram realizados no estado de São Paulo, nos municípios de Mogi das Cruzes, Jarinu, Piracicaba e Itatiba, nos anos de 2000 e 2001, com o objetivo de avaliar os efeitos da solarização nas propriedades físicas, químicas e biológicas dos solos. A solarização reduziu significativamente a resistência à penetração dos solos nos ensaios de Jarinu, Piracicaba e Itatiba. Em Jarinu, oito meses após a retirada do plástico, as diferenças entre os tratamentos permaneceram. Por outro lado, em Mogi das Cruzes, onde o ensaio foi instalado em solo turfoso, a solarização causou aumento na resistência na camada de 2,5 a 5 cm de profundidade. Nos ensaios de Piracicaba e Jarinu, foram feitas avaliações de macro, microporosidade, porosidade total e densidade, não tendo os tratamentos diferido entre si, porém houve uma tendência de redução na densidade dos solos solarizados. A atividade microbiana, avaliada pela hidrólise de diacetato de fluoresceína, foi reduzida pela solarização. A supressividade a Fusarium oxysporum f. sp. phaseoli foi avaliada in vitro, pela colonização de amostras de solo usando um isolado marcado com resistência a benomyl, no ensaio de Mogi das Cruzes. A solarização reduziu a recuperação do patógeno, evidenciando um aumento da supressividade. Nos solos solarizados, houve aumento significativo dos teores de N-NH4+ em todos os experimentos, Mn em três, N-NO3-, Mg2+ e saturação por bases em dois e K+ em um experimento. Ocorreu redução dos teores de Cu, Fe e H + Al em dois experimentos e Zn em um ensaio. Segundo os resultados, a solarização promoveu alterações nas propriedades físicas, químicas e biológicas dos solos, melhorando a estrutura, liberando nutrientes e

  2. Evaluación agronómica de líneas avanzadas de fríjol voluble Phaseolus vulgaris L. en Paipa, Boyacá

    Directory of Open Access Journals (Sweden)

    Diana Katherine Ríos

    2014-06-01

    Full Text Available En Boyacá la oferta de germoplasma de frijol mejorado es baja. El objetivo de este estudio fue evaluar el comportamiento agronómico de 18 líneas avanzadas volubles de frijol, procedentes del CIAT. Se utilizó el diseño bloques completos al azar, con tres repeticiones, incluyendo Bola Roja, Bolívar y Cargamanto como testigos. Se evaluó días a floración, vigor de crecimiento, días a madurez, incidencia de enfermedades, número de vainas/planta, número de granos/vaina, peso de 100 granos y rendimiento. Los genotipos MAC27, MAC13, MBC26, MBC28 y MBC37 no se diferenciaron del Cargamanto pero sí mostraron ser más precoces que Bola Roja y Bolívar. En días a madurez MAC27 y MBC39 fueron más tempranos que los testigos. Todos los genotipos se comportaron como resistentes a enfermedades, excepto MBC30 que resultó susceptible a Uromyces phaseoli y MBC33 a Phoma exigua. En número de vainas por planta, número de granos por vaina, peso de 100 semillas y rendimiento se destacaron MAC27 y LAS399, sin diferencia significativa frente a Cargamanto pero si superando a Bola Roja y Bolívar. Pese al efecto de las heladas en la fase final del ciclo, LAS399, MAC27 y Cargamanto produjeron rendimientos de 1300, 1038 y 950 kg/ha, respectivamente, mostrando las ventajas de su precocidad, en comparación a Bola Roja y Bolívar que son tardíos. Se concluye que LAS399 y MAC27, por su alta precocidad y resistencia a patógenos, son las líneas más promisorias para continuar la evaluación respecto a la posibilidad de ampliar la oferta de germoplasma de frijol voluble en Boyacá.

  3. Complete genome sequencing of Agrobacterium sp. H13-3, the former Rhizobium lupini H13-3, reveals a tripartite genome consisting of a circular and a linear chromosome and an accessory plasmid but lacking a tumor-inducing Ti-plasmid.

    Science.gov (United States)

    Wibberg, Daniel; Blom, Jochen; Jaenicke, Sebastian; Kollin, Florian; Rupp, Oliver; Scharf, Birgit; Schneiker-Bekel, Susanne; Sczcepanowski, Rafael; Goesmann, Alexander; Setubal, Joao Carlos; Schmitt, Rüdiger; Pühler, Alfred; Schlüter, Andreas

    2011-08-20

    Agrobacterium sp. H13-3, formerly known as Rhizobium lupini H13-3, is a soil bacterium that was isolated from the rhizosphere of Lupinus luteus. The isolate has been established as a model system for studying novel features of flagellum structure, motility and chemotaxis within the family Rhizobiaceae. The complete genome sequence of Agrobacterium sp. H13-3 has been established and the genome structure and phylogenetic assignment of the organism was analysed. For de novo sequencing of the Agrobacterium sp. H13-3 genome, a combined strategy comprising 454-pyrosequencing on the Genome Sequencer FLX platform and PCR-based amplicon sequencing for gap closure was applied. The finished genome consists of three replicons and comprises 5,573,770 bases. Based on phylogenetic analyses, the isolate could be assigned to the genus Agrobacterium biovar I and represents a genomic species G1 strain within this biovariety. The highly conserved circular chromosome (2.82 Mb) of Agrobacterium sp. H13-3 mainly encodes housekeeping functions characteristic for an aerobic, heterotrophic bacterium. Agrobacterium sp. H13-3 is a motile bacterium driven by the rotation of several complex flagella. Its behaviour towards external stimuli is regulated by a large chemotaxis regulon and a total of 17 chemoreceptors. Comparable to the genome of Agrobacterium tumefaciens C58, Agrobacterium sp. H13-3 possesses a linear chromosome (2.15 Mb) that is related to its reference replicon and features chromosomal and plasmid-like properties. The accessory plasmid pAspH13-3a (0.6 Mb) is only distantly related to the plasmid pAtC58 of A. tumefaciens C58 and shows a mosaic structure. A tumor-inducing Ti-plasmid is missing in the sequenced strain H13-3 indicating that it is a non-virulent isolate. Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Linear Chromosome-generating System of Agrobacterium tumefaciens C58: Protelomerase Generates and Protects Hairpin Ends

    Energy Technology Data Exchange (ETDEWEB)

    Huang, Wai Mun; DaGloria, Jeanne; Fox, Heather; Ruan, Qiurong; Tillou, John; Shi, Ke; Aihara, Hideki; Aron, John; Casjens, Sherwood (Utah); (UMM)

    2012-09-05

    Agrobacterium tumefaciens C58, the pathogenic bacteria that causes crown gall disease in plants, harbors one circular and one linear chromosome and two circular plasmids. The telomeres of its unusual linear chromosome are covalently closed hairpins. The circular and linear chromosomes co-segregate and are stably maintained in the organism. We have determined the sequence of the two ends of the linear chromosome thus completing the previously published genome sequence of A. tumefaciens C58. We found that the telomeres carry nearly identical 25-bp sequences at the hairpin ends that are related by dyad symmetry. We further showed that its Atu2523 gene encodes a protelomerase (resolvase) and that the purified enzyme can generate the linear chromosomal closed hairpin ends in a sequence-specific manner. Agrobacterium protelomerase, whose presence is apparently limited to biovar 1 strains, acts via a cleavage-and-religation mechanism by making a pair of transient staggered nicks invariably at 6-bp spacing as the reaction intermediate. The enzyme can be significantly shortened at both the N and C termini and still maintain its enzymatic activity. Although the full-length enzyme can uniquely bind to its product telomeres, the N-terminal truncations cannot. The target site can also be shortened from the native 50-bp inverted repeat to 26 bp; thus, the Agrobacterium hairpin-generating system represents the most compact activity of all hairpin linear chromosome- and plasmid-generating systems to date. The biochemical analyses of the protelomerase reactions further revealed that the tip of the hairpin telomere may be unusually polymorphically capable of accommodating any nucleotide.

  5. Single acquisition of protelomerase gave rise to speciation of a large and diverse clade within the Agrobacterium/Rhizobium supercluster characterized by the presence of a linear chromid.

    Science.gov (United States)

    Ramírez-Bahena, Martha H; Vial, Ludovic; Lassalle, Florent; Diel, Benjamin; Chapulliot, David; Daubin, Vincent; Nesme, Xavier; Muller, Daniel

    2014-04-01

    Linear chromosomes are atypical in bacteria and likely a secondary trait derived from ancestral circular molecules. Within the Rhizobiaceae family, whose genome contains at least two chromosomes, a particularity of Agrobacterium fabrum (formerly A. tumefaciens) secondary chromosome (chromid) is to be linear and hairpin-ended thanks to the TelA protelomerase. Linear topology and telA distributions within this bacterial family was screened by pulse field gel electrophoresis and PCR. In A. rubi, A. larrymoorei, Rhizobium skierniewicense, A. viscosum, Agrobacterium sp. NCPPB 1650, and every genomospecies of the biovar 1/A. tumefaciens species complex (including R. pusense, A. radiobacter, A. fabrum, R. nepotum plus seven other unnamed genomospecies), linear chromid topologies were retrieved concomitantly with telA presence, whereas the remote species A. vitis, Allorhizobium undicola, Rhizobium rhizogenes and Ensifer meliloti harbored a circular chromid as well as no telA gene. Moreover, the telA phylogeny is congruent with that of recA used as a marker gene of the Agrobacterium phylogeny. Collectively, these findings strongly suggest that single acquisition of telA by an ancestor was the founding event of a large and diverse clade characterized by the presence of a linear chromid. This clade, characterized by unusual genome architecture, appears to be a relevant candidate to serve as a basis for a possible redefinition of the controversial Agrobacterium genus. In this respect, investigating telA in sequenced genomes allows to both ascertain the place of concerned strains into Agrobacterium spp. and their actual assignation to species/genomospecies in this genus. Copyright © 2014 Elsevier Inc. All rights reserved.

  6. Characterization and protective property of Brucella abortus cydC and looP mutants.

    Science.gov (United States)

    Truong, Quang Lam; Cho, Youngjae; Barate, Abhijit Kashinath; Kim, Suk; Hahn, Tae-Wook

    2014-11-01

    Brucella abortus readily multiplies in professional or nonprofessional phagocytes in vitro and is highly virulent in mice. Isogenic mutants of B. abortus biovar 1 strain IVKB9007 lacking the ATP/GDP-binding protein motif A (P-loop) (named looP; designated here the IVKB9007 looP::Tn5 mutant) and the ATP-binding/permease protein (cydC; designated here the IVKB9007 cydC::Tn5 mutant) were identified and characterized by transposon mutagenesis using the mini-Tn5Km2 transposon. Both mutants were found to be virtually incapable of intracellular replication in both murine macrophages (RAW264.7) and the HeLa cell line, and their virulence was significantly impaired in BALB/c mice. Respective complementation of the IVKB9007 looP::Tn5 and IVKB9007 cydC::Tn5 mutants restored their ability to survive in vitro and in vivo to a level comparable with that of the wild type. These findings indicate that the cydC and looP genes play important roles in the virulence of B. abortus. In addition, intraperitoneal immunization of mice with a dose of the live IVKB9007 looP::Tn5 and IVKB9007 cydC::Tn5 mutants provided a high degree of protection against challenge with pathogenic B. abortus strain 544. Both mutants should be evaluated further as a live attenuated vaccine against bovine brucellosis for their ability to stimulate a protective immune response. Copyright © 2014, American Society for Microbiology. All Rights Reserved.

  7. Review of Brucellosis in Nepal

    Science.gov (United States)

    Acharya, Krishna Prasad; Niroula, Nirajan; Kaphle, Krishna

    2016-01-01

    OBJECTIVES The aim of this paper is to evaluate the current status of the disease, the mechanism of infection, and pathogenesis, its zoonotic potential, diagnostic advances, treatment regimens, and the preventive measures that can be adopted in managing human brucellosis in under-developed countries such as Nepal. METHODS We performed a systematic review of all the available literture through Google Scholar, PubMed, Gideon Informatics, World Health Organization and other legitimate sources. Other secondary informations were collected from the government agencies such as department of livestock services and Ministry of Health. The obtained information was then re-analysed and summarized. RESULTS Few publications have addressed brucellosis in Nepal and most of those publications have focused on bovine brucellosis with sparse information available on brucellosis in humans and small ruminants. Brucella abortus is the most predominant causative agent followed by B. suis. B. abortus is predominant in cattle accounting for a substantial portion of bovine abortion in the country. Lack of awareness, unhealthy food habit, traditional husbandry practices, and a lack of surveillance and immunization have been the major factors in maintaining a vicious cycle of propagation of the disease in human and animals. Unfortunately, nothing has been done to identify the species of Brucella at the biovar level. CONCLUSIONS Although brucellosis has been reported to be endemic in Nepal, neither the distribution nor the economic and public health impact of this disease is well characterized. Robust and well-designed nationwide survey is warranted to assess the prevalence and distribution of disease in livestock and humans. Such data would facilitate the design of appropriate control programmes. PMID:27703129

  8. Genome-scale reconstruction of the metabolic network in Yersinia pestis, strain 91001

    Energy Technology Data Exchange (ETDEWEB)

    Navid, A; Almaas, E

    2009-01-13

    The gram-negative bacterium Yersinia pestis, the aetiological agent of bubonic plague, is one the deadliest pathogens known to man. Despite its historical reputation, plague is a modern disease which annually afflicts thousands of people. Public safety considerations greatly limit clinical experimentation on this organism and thus development of theoretical tools to analyze the capabilities of this pathogen is of utmost importance. Here, we report the first genome-scale metabolic model of Yersinia pestis biovar Mediaevalis based both on its recently annotated genome, and physiological and biochemical data from literature. Our model demonstrates excellent agreement with Y. pestis known metabolic needs and capabilities. Since Y. pestis is a meiotrophic organism, we have developed CryptFind, a systematic approach to identify all candidate cryptic genes responsible for known and theoretical meiotrophic phenomena. In addition to uncovering every known cryptic gene for Y. pestis, our analysis of the rhamnose fermentation pathway suggests that betB is the responsible cryptic gene. Despite all of our medical advances, we still do not have a vaccine for bubonic plague. Recent discoveries of antibiotic resistant strains of Yersinia pestis coupled with the threat of plague being used as a bioterrorism weapon compel us to develop new tools for studying the physiology of this deadly pathogen. Using our theoretical model, we can study the cell's phenotypic behavior under different circumstances and identify metabolic weaknesses which may be harnessed for the development of therapeutics. Additionally, the automatic identification of cryptic genes expands the usage of genomic data for pharmaceutical purposes.

  9. Efficacy of standard therapies against Ureaplasma species and persistence among men with non-gonococcal urethritis enrolled in a randomized controlled trial

    Science.gov (United States)

    Khosropour, Christine M.; Manhart, Lisa E.; Gillespie, Catherine W.; Lowens, M. Sylvan; Golden, Matthew R.; Jensen, Nicole L.; Kenny, George E.; Totten, Patricia A.

    2015-01-01

    Objective U. urealyticum biovar 2 (UU-2) but not U. parvum (UP) has been associated with non-gonococcal urethritis (NGU), but little is known about species-specific responses to standard therapies. We examined species-specific treatment outcomes and followed men with treatment failure for 9 weeks. Methods From May 2007-July 2011, men aged ≥16 attending an STD clinic in Seattle, Washington with NGU (urethral discharge or urethral symptoms plus ≥5 PMNs/HPF) were enrolled in a double-blind, randomized trial. Participants received active azithromycin (1g) + placebo doxycycline or active doxycycline (100mg bid × 7d) + placebo azithromycin. Ureaplasmas were detected in culture followed by species-specific PCR. Outcomes were assessed at 3, 6, and 9 weeks. At 3 weeks, men with persistent Ureaplasmas received “reverse therapy” (e.g., active doxycycline if they first received active azithromycin). At 6 weeks, persistently-positive men received moxifloxacin (400mg × 7d). Results Of 490 men, 107 (22%) and 60 (12%) were infected with UU-2 and UP, respectively, and returned at 3 weeks. Persistent infection was similar for UU-2-infected men initially treated with azithromycin or doxycycline (25% vs. 31%, P=0.53), but differed somewhat for men with UP (45% vs. 24%; P=0.11). At 6 weeks, 57% of UU-2-infected and 63% of UP-infected men who received both drugs had persistent infection. Failure after moxifloxacin occurred in 30% and 36%, respectively. Persistent detection of UU-2 or UP was not associated with signs/symptoms of NGU. Conclusion Persistent infection after treatment with doxycycline, azithromycin, and moxifloxacin was common for UU and UP, but not associated with persistent urethritis. PMID:25616607

  10. Suboptimal adherence to doxycycline and treatment outcomes among men with non-gonococcal urethritis: a prospective cohort study.

    Science.gov (United States)

    Khosropour, Christine M; Manhart, Lisa E; Colombara, Danny V; Gillespie, Catherine W; Lowens, M Sylvan; Totten, Patricia A; Golden, Matthew R; Simoni, Jane

    2014-02-01

    Doxycycline, one of two recommended therapies for non-gonococcal urethritis (NGU), consists of a 7-day course of therapy (100 mg BID). Since suboptimal adherence may contribute to poor treatment outcomes, we examined the association between self-reported imperfect adherence to doxycycline and clinical and microbiologic failure among men with NGU. Men aged ≥16 years with NGU attending a Seattle, WA, sexually transmitted diseases clinic were enrolled in a double-blind, parallel-group superiority trial from January 2007 to July 2011. Men were randomised to active doxycycline/placebo azithromycin or placebo doxycycline/active azithromycin. Imperfect adherence was defined as missing ≥1 dose in 7 days. Urine was tested for Chlamydia trachomatis (CT), Mycoplasma genitalium (MG), and Ureaplasma urealyticum-biovar 2 (UU-2) using nucleic acid amplification tests. Clinical failure (symptoms and ≥5 PMNs/HPF or discharge) and microbiologic failure (positive tests for CT, MG, and/or UU-2) were determined after 3 weeks. 184 men with NGU were randomised to active doxycycline and provided data on adherence. Baseline prevalence of CT, MG and UU-2 was 26%, 13% and 27%, respectively. 28% of men reported imperfect adherence, and this was associated with microbiologic failure among men with CT (aRR=9.33; 95% CI 1.00 to 89.2) and UU-2 (aRR=3.08; 95% CI 1.31 to 7.26) but not MG. Imperfect adherence was not significantly associated with clinical failure overall or for any specific pathogens, but it was more common among imperfectly adherent men with CT (aRR=2.63; 0.93-7.41, p=0.07). Adherence may be important for microbiologic cure of select pathogens. Factors other than adherence should be considered for CT-negative men with persistent NGU.

  11. Standard treatment regimens for nongonococcal urethritis have similar but declining cure rates: a randomized controlled trial.

    Science.gov (United States)

    Manhart, Lisa E; Gillespie, Catherine W; Lowens, M Sylvan; Khosropour, Christine M; Colombara, Danny V; Golden, Matthew R; Hakhu, Navneet R; Thomas, Katherine K; Hughes, James P; Jensen, Nicole L; Totten, Patricia A

    2013-04-01

    Azithromycin or doxycycline is recommended for nongonococcal urethritis (NGU); recent evidence suggests their efficacy has declined. We compared azithromycin and doxycycline in men with NGU, hypothesizing that azithromycin was more effective than doxycycline. From January 2007 to July 2011, English-speaking males ≥16 years, attending a sexually transmitted diseases clinic in Seattle, Washington, with NGU (visible urethral discharge or ≥5 polymorphonuclear leukocytes per high-power field [PMNs/HPF]) were eligible for this double-blind, parallel-group superiority trial. Participants received active azithromycin (1 g) + placebo doxycycline or active doxycycline (100 mg twice daily for 7 days) + placebo azithromycin. Urine was tested for Chlamydia trachomatis (CT), Mycoplasma genitalium (MG), Ureaplasma urealyticum biovar 2 (UU-2), and Trichomonas vaginalis (TV) using nucleic acid amplification tests. Clinical cure (urethral symptoms and absence of discharge) and microbiologic cure (negative tests for CT, MG, and/or UU-2) were determined after 3 weeks. Of 606 men, 304 were randomized to azithromycin and 302 to doxycycline; CT, MG, TV, and UU-2 were detected in 24%, 13%, 2%, and 23%, respectively. In modified intent-to-treat analyses, 172 of 216 (80%; 95% confidence interval [CI], 74%-85%) receiving azithromycin and 157 of 206 (76%; 95% CI, 70%-82%) receiving doxycycline experienced clinical cure (P = .40). In pathogen-specific analyses, clinical cure did not differ by arm, nor did microbiologic cure differ for CT (86% vs 90%, P = .56), MG (40% vs 30%, P = .41), or UU-2 (75% vs 70%, P = .50). No unexpected adverse events occurred. Clinical and microbiologic cure rates for NGU were somewhat low and there was no significant difference between azithromycin and doxycycline. Mycoplasma genitalium treatment failure was extremely common. Clinical Trials Registration.NCT00358462.

  12. Sub-optimal adherence to doxycycline and treatment outcomes among men with non-gonococcal urethritis: a prospective cohort study

    Science.gov (United States)

    Khosropour, Christine M.; Manhart, Lisa E.; Colombara, Danny V.; Gillespie, Catherine W.; Lowens, M. Sylvan; Totten, Patricia A.; Golden, Matthew R.; Simoni, Jane

    2014-01-01

    Objective Doxycycline, one of two recommended therapies for non-gonococcal urethritis (NGU), consists of a seven-day course of therapy (100mg BID). Since suboptimal adherence may contribute to poor treatment outcomes, we examined the association between self-reported imperfect adherence to doxycycline and clinical and microbiologic failure among men with NGU. Methods Men aged ≥16 years with NGU attending a Seattle, WA sexually transmitted diseases clinic were enrolled in a double-blind, parallel-group superiority trial from January 2007 to July 2011. Men were randomized to active doxycycline/placebo azithromycin or placebo doxycycline/active azithromycin. Imperfect adherence was defined as missing ≥1 dose in 7 days. Urine was tested for Chlamydia trachomatis (CT), Mycoplasma genitalium (MG), and Ureaplasma urealyticum-biovar2 (UU-2) using nucleic acid amplification tests. Clinical failure (symptoms and ≥5 PMNs/HPF or discharge) and microbiologic failure (positive tests for CT, MG, and/or UU-2) were determined after 3-weeks. Results 184 men with NGU were randomized to active doxycycline and provided data on adherence. Baseline prevalence of CT, MG, and UU-2 was 26%, 13%, and 27%, respectively. 28% of men reported imperfect adherence and this was associated with microbiologic failure among men with CT (aRR=9.33; 95% CI=1.00–89.2) and UU-2 (aRR=3.08; 95% CI=1.31–7.26) but not MG. Imperfect adherence was not significantly associated with clinical failure overall or for any specific pathogens, but it was more common among imperfectly adherent men with CT (aRR=2.63; 0.93–7.41, p=0.07). Conclusions Adherence may be important for microbiologic cure of select pathogens. Factors other than adherence should be considered for CT-negative men with persistent NGU. PMID:24106340

  13. Attenuated enzootic (pestoides) isolates of Yersinia pestis express active aspartase.

    Science.gov (United States)

    Bearden, Scott W; Sexton, Christopher; Pare, Joshua; Fowler, Janet M; Arvidson, Cindy G; Yerman, Lyudmyla; Viola, Ronald E; Brubaker, Robert R

    2009-01-01

    It is established that Yersinia pestis, the causative agent of bubonic plague, recently evolved from enteropathogenic Yersinia pseudotuberculosis by undergoing chromosomal degeneration while acquiring two unique plasmids that facilitate tissue invasion (pPCP) and dissemination by fleabite (pMT). Thereafter, plague bacilli spread from central Asia to sylvatic foci throughout the world. These epidemic isolates exhibit a broad host range including man as opposed to enzootic (pestoides) variants that remain in ancient reservoirs where infection is limited to muroid rodents. Cells of Y. pseudotuberculosis are known to express glucose-6-phosphate dehydrogenase (Zwf) and aspartase (AspA); these activities are not detectable in epidemic Y. pestis due to missense mutations (substitution of proline for serine at amino position 155 of Zwf and leucine for valine at position 363 of AspA). In this study, functional Zwf was found in pestoides strains E, F and G but not seven other enzootic isolates; enzymic activity was associated with retention of serine at amino acid position 155. Essentially, full AspA activity occurred in pestoides isolates where valine (pestoides A, B, C and D) or serine (pestoides E, F, G and I) occupied position 363. Reduced activity occurred in strains Angola and A16, which contained phenylalanine at this position. The kcat but not Km of purified AspA from strain Angola was significantly reduced. In this context, aspA of the recently described attenuated enzootic microtus biovar encodes active valine at position 363, further indicating that functional AspA is a biomarker for avirulence of Y. pestis in man.

  14. Antibody Reactivity to Omp31 from Brucella melitensis in Human and Animal Infections by Smooth and Rough Brucellae

    Science.gov (United States)

    Cassataro, Juliana; Pasquevich, Karina; Bruno, Laura; Wallach, Jorge C.; Fossati, Carlos A.; Baldi, Pablo C.

    2004-01-01

    Group 3 of outer membrane proteins (OMPs) of Brucella includes Omp25 and Omp31, which share 34% identity. Omp25 is highly conserved in Brucella species, and Omp31 is present in all Brucella species, except Brucella abortus. Antibodies to Brucella melitensis Omp31 have been sought only in infected sheep, and Western blotting of sera from infected sheep did not reveal anti-Omp31 reactivity. We obtained recombinant purified Omp31 (B. melitensis) and tested its recognition by sera from humans and animals suffering from brucellosis by an indirect enzyme-linked immunosorbent assay (ELISA). Serum samples from 74 patients, 57 sheep, and 47 dogs were analyzed; brucellosis was confirmed by bacteriological isolation in all ovine and canine cases and 31 human cases of brucellosis. Thirty-five patients (47%) were positive for antibodies to Omp31, including seven cases of Brucella suis infection, two cases of B. abortus infection, and three cases of B. melitensis infection. Of 39 sheep naturally infected with B. melitensis (biovars 1 and 3), 23 (59%) were positive for antibodies to Omp31. Anti-Omp31 antibodies were also detected in 12 of 18 rams (67%) in which Brucella ovis was isolated from semen. Antibodies to Omp31 were also found in 41 (87%) of the 47 dogs, including 13 with recent infection. These results suggest that an indirect ELISA using recombinant purified Omp31 from B. melitensis would be of limited value for the diagnosis of human and animal brucellosis. Nevertheless, the potential usefulness of this antigen in combination with other recombinant proteins from Brucella should not be dismissed.   PMID:14715555

  15. Penicillin kills Chlamydia following the fusion of bacteria with lysosomes and prevents genital inflammatory lesions in C. muridarum-infected mice.

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    Maud Dumoux

    Full Text Available The obligate intracellular bacterium Chlamydia exists as two distinct forms. Elementary bodies (EBs are infectious and extra-cellular, whereas reticulate bodies (RBs replicate within a specialized intracellular compartment termed an 'inclusion'. Alternative persistent intra-cellular forms can be induced in culture by diverse stimuli such as IFNγ or adenosine/EHNA. They do not grow or divide but revive upon withdrawal of the stimulus and are implicated in several widespread human diseases through ill-defined in vivo mechanisms. β-Lactam antibiotics have also been claimed to induce persistence in vitro. The present report shows that upon penicillin G (pG treatment, inclusions grow as fast as those in infected control cells. After removal of pG, Chlamydia do not revert to RBs. These effects are independent of host cell type, serovar, biovar and species of Chlamydia. Time-course experiments demonstrated that only RBs were susceptible to pG. pG-treated bacteria lost their control over host cell apoptotic pathways and no longer expressed pre-16S rRNA, in contrast to persistent bacteria induced with adenosine/EHNA. Confocal and live-video microscopy showed that bacteria within the inclusion fused with lysosomal compartments in pG-treated cells. That leads to recruitment of cathepsin D as early as 3 h post pG treatment, an event preceding bacterial death by several hours. These data demonstrate that pG treatment of cultured cells infected with Chlamydia results in the degradation of the bacteria. In addition we show that pG is significantly more efficient than doxycycline at preventing genital inflammatory lesions in C. muridarum-C57Bl/6 infected mice. These in vivo results support the physiological relevance of our findings and their potential therapeutic applications.

  16. Novel plasmids and resistance phenotypes in Yersinia pestis: unique plasmid inventory of strain Java 9 mediates high levels of arsenic resistance.

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    Eppinger, Mark; Radnedge, Lyndsay; Andersen, Gary; Vietri, Nicholas; Severson, Grant; Mou, Sherry; Ravel, Jacques; Worsham, Patricia L

    2012-01-01

    Growing evidence suggests that the plasmid repertoire of Yersinia pestis is not restricted to the three classical virulence plasmids. The Java 9 strain of Y. pestis is a biovar Orientalis isolate obtained from a rat in Indonesia. Although it lacks the Y. pestis-specific plasmid pMT, which encodes the F1 capsule, it retains virulence in mouse and non-human primate animal models. While comparing diverse Y. pestis strains using subtractive hybridization, we identified sequences in Java 9 that were homologous to a Y. enterocolitica strain carrying the transposon Tn2502, which is known to encode arsenic resistance. Here we demonstrate that Java 9 exhibits high levels of arsenic and arsenite resistance mediated by a novel promiscuous class II transposon, named Tn2503. Arsenic resistance was self-transmissible from Java 9 to other Y. pestis strains via conjugation. Genomic analysis of the atypical plasmid inventory of Java 9 identified pCD and pPCP plasmids of atypical size and two previously uncharacterized cryptic plasmids. Unlike the Tn2502-mediated arsenic resistance encoded on the Y. enterocolitica virulence plasmid; the resistance loci in Java 9 are found on all four indigenous plasmids, including the two novel cryptic plasmids. This unique mobilome introduces more than 105 genes into the species gene pool. The majority of these are encoded by the two entirely novel self-transmissible plasmids, which show partial homology and synteny to other enterics. In contrast to the reductive evolution in Y. pestis, this study underlines the major impact of a dynamic mobilome and lateral acquisition in the genome evolution of the plague bacterium.

  17. Novel plasmids and resistance phenotypes in Yersinia pestis: unique plasmid inventory of strain Java 9 mediates high levels of arsenic resistance.

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    Mark Eppinger

    Full Text Available Growing evidence suggests that the plasmid repertoire of Yersinia pestis is not restricted to the three classical virulence plasmids. The Java 9 strain of Y. pestis is a biovar Orientalis isolate obtained from a rat in Indonesia. Although it lacks the Y. pestis-specific plasmid pMT, which encodes the F1 capsule, it retains virulence in mouse and non-human primate animal models. While comparing diverse Y. pestis strains using subtractive hybridization, we identified sequences in Java 9 that were homologous to a Y. enterocolitica strain carrying the transposon Tn2502, which is known to encode arsenic resistance. Here we demonstrate that Java 9 exhibits high levels of arsenic and arsenite resistance mediated by a novel promiscuous class II transposon, named Tn2503. Arsenic resistance was self-transmissible from Java 9 to other Y. pestis strains via conjugation. Genomic analysis of the atypical plasmid inventory of Java 9 identified pCD and pPCP plasmids of atypical size and two previously uncharacterized cryptic plasmids. Unlike the Tn2502-mediated arsenic resistance encoded on the Y. enterocolitica virulence plasmid; the resistance loci in Java 9 are found on all four indigenous plasmids, including the two novel cryptic plasmids. This unique mobilome introduces more than 105 genes into the species gene pool. The majority of these are encoded by the two entirely novel self-transmissible plasmids, which show partial homology and synteny to other enterics. In contrast to the reductive evolution in Y. pestis, this study underlines the major impact of a dynamic mobilome and lateral acquisition in the genome evolution of the plague bacterium.

  18. Epidemiology of Brucella infection in the human, livestock and wildlife interface in the Katavi-Rukwa ecosystem, Tanzania.

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    Assenga, Justine A; Matemba, Lucas E; Muller, Shabani K; Malakalinga, Joseph J; Kazwala, Rudovick R

    2015-08-08

    Brucellosis is a zoonosis of public health importance worldwide. In Tanzania, the disease is underreported due to insufficient awareness, inadequate diagnostic protocols, including lack of appropriate reagents for diagnosis. Livestock and wildlife are considered potential sources of infection to humans; however, the role played by these carriers in the epidemiology of the disease in the ecosystems in Tanzania is not fully understood. The objective of this study was to establish the prevalence of anti-Brucella antibodies in humans, wildlife and livestock; and molecular prevalence of Brucella spp in cattle and goats in the Katavi- Rukwa ecosystem. Anti-Brucella antibodies were detected in humans at 0.6 % (95 % CI: 0.1, 2.1 %); cattle at 6.8 % (95 % CI: 5.4, 8.5 %), goats at 1.6 % (95 % CI: 0.4, 4.1 %) and buffaloes at 7.9 % (95 % CI: 1.7, 21.4 %). One of the two sampled lions tested positive. Cattle had a significantly higher prevalence of anti-Brucella antibodies as compared to goats (P Brucella infection. Eight (3.5 %) out of 231 milk samples tested were positive for Brucella spp on Polymerase Chain Reaction (PCR), and Brucella abortus biovar 1 was detected in cattle milk. However, no Brucella spp were detected in goat milk. This study has shown the presence of anti- Brucella antibodies in humans, livestock, and wildlife in the Katavi- Rukwa ecosystem. Transmission of the infection between wildlife, livestock and humans is likely to continue due to increasing human activities in the human wildlife interface. This information is an important contribution to public health policy development in the human wildlife interface of the Katavi- Rukwa ecosystem.

  19. Different resistance patterns of reference and field strains of Brucella abortus

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    Karina L. Miranda

    2015-03-01

    Full Text Available The aim of this study was to evaluate the growth of the B. abortus reference strains and field isolates on media containing different inhibitor agents. Reference strains were seeded on tryptose agar containing: i-erythritol (1.0 mg/mL, fuchsin (20 μg/mL and 80 μg/mL, thionin (2.5 μg/mL and 10 μg/mL, rifampicin (200 μg/mL and safranin O (200 μg/mL. Field isolates were tested only on media containing i-erythritol, rifampicin and thionin. Furthermore, each suspension was also inoculated on tryptose agar incubated in air, to test its ability to grow without CO2. Sensitivity to fuchsin was similar among reference strains evaluated. Growth of S19, 544 and 2308 but not RB51 were inhibited on media containing rifampicin. Medium with safranin O showed no inhibition for RB51, 544 and 2308, but it partially inhibited the S19 growth as well as medium containing i-erythritol. Treatment/control growth ratio for 2308 on tryptose agar containing thionin (2.5 μg/mL was approximatelly 1.0, whereas S19 and RB51 showed 0.85 and 0.89 ratios, respectively. Growth of 544, S19 and RB51 but not 2308 was completely inhibited on medium with thionin (10 μg/mL. All field strains grew on medium containing i-erythritol, but were completelly inhibited by rifampicin. With exception of A1 (B. abortus biovar 3 all field isolates grew on medium with thionin, although some strains showed a treatment/control growth ratio of 0.75–0.80 (10 μg/mL. These results showed that tryptose agar with thionin, i-erythritol or rifampicin could be useful for differentiating vaccine, challenge and field strains of B. abortus.

  20. Molecular and Epidemiological Review of Toxigenic Diphtheria Infections in England between 2007 and 2013

    Science.gov (United States)

    Both, Leonard; Collins, Sarah; de Zoysa, Aruni; White, Joanne; Mandal, Sema

    2014-01-01

    Human infections caused by toxigenic corynebacteria occur sporadically across Europe. In this report, we undertook the epidemiological and molecular characterization of all toxigenic corynebacterium strains isolated in England between January 2007 and December 2013. Epidemiological aspects include case demographics, risk factors, clinical presentation, treatment, and outcome. Molecular characterization was performed using multilocus sequence typing (MLST) alongside traditional phenotypic methods. In total, there were 20 cases of toxigenic corynebacteria; 12 (60.0%) were caused by Corynebacterium ulcerans, where animal contact was the predominant risk factor. The remaining eight (40.0%) were caused by Corynebacterium diphtheriae strains; six were biovar mitis, which were associated with recent travel abroad. Adults 45 years and older were particularly affected (55.0%; 11/20), and typical symptoms included sore throat and fever. Respiratory diphtheria with the absence of a pharyngeal membrane was the most common presentation (50.0%; 10/20). None of the eight C. diphtheriae cases were fully immunized. Diphtheria antitoxin was issued in two (9.5%) cases; both survived. Two (9.5%) cases died, one due to a C. diphtheriae infection and one due to C. ulcerans. MLST demonstrated that the majority (87.5%; 7/8) of C. diphtheriae strains represented new sequence types (STs). By adapting several primer sequences, the MLST genes in C. ulcerans were also amplified, thereby providing the basis for extension of the MLST scheme, which is currently restricted to C. diphtheriae. Despite high population immunity, occasional toxigenic corynebacterium strains are identified in England and continued surveillance is required. PMID:25502525

  1. Characterization of Probiotic Fermented Milk Prepared by Different Inoculation Size of Mesophilic and Thermophilic Lactic Acid Bacteria

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    Sara Nasiri Boosjin

    2016-10-01

    Full Text Available Background and Objectives: Importance of development of novel probiotic fermented milk and challenge made for its acceptability is well known. In this research, the impact of different inoculation sizes of yogurt and DL-type starter culture (mesophilic and thermophilic LAB on titratable acidity, viscosity, sensorial and microbial properties of fermented milk was investigated; and finally, probiotic Langfil was produced.Materials and Methods: Fermented milk produced by 1, 2 and 3% v v-1 inocula consisting thermophilic: mesophilic starter cultures 10:90 (Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. cremoris, Lactococcus lactis subsp. lactis, Lactococcus lactis subsp. lactis biovar. diacetylactis and Leuconostoc mesenteroides subsp. cremoris. Streptococcus thermophilus and Lactobacillus delbrueckii subsp. Bulgaricus were analyzed for determination of titratable acidity, viscosity, viability of mesophilic starter cultures and sensory properties on days 5, 10, and 15 of storage at 4°C. Then, the most suitable treatments were selected for the producing probiotic Langfil, containing probiotic starter culture (2% v v-1 inoculums with equal ratio of Lactobacillus acidophilus and Bifidobacterium bifidum. Lactococcus lactis and L. cremoris were counted on M17 agar, while Leuconostoc and Lactobacillus were counted aerobically on tomato juice agar and MRS bile agar, respectively. Bifidobacterium was cultured anaerobically on MRS bile agar. Sensory evaluation was carried out by ten trained panelists, based on a nine-point hedonic scale during the cold storage.Results and Conclusion: According to results, the best organoleptic properties were achieved in the product prepared with 2% the mesophilic and thermophilic starter cultures and 2% probiotic. This product had a high viscosity. An Iranian probiotic Langfil with desired properties was produced using the best treatment prepared.Conflict of interests: The authors declare no conflict of

  2. The Vibrio cholerae var regulon encodes a metallo-β-lactamase and an antibiotic efflux pump, which are regulated by VarR, a LysR-type transcription factor.

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    Hong-Ting Victor Lin

    Full Text Available The genome sequence of V. cholerae O1 Biovar Eltor strain N16961 has revealed a putative antibiotic resistance (var regulon that is predicted to encode a transcriptional activator (VarR, which is divergently transcribed relative to the putative resistance genes for both a metallo-β-lactamase (VarG and an antibiotic efflux-pump (VarABCDEF. We sought to test whether these genes could confer antibiotic resistance and are organised as a regulon under the control of VarR. VarG was overexpressed and purified and shown to have β-lactamase activity against penicillins, cephalosporins and carbapenems, having the highest activity against meropenem. The expression of VarABCDEF in the Escherichia coli (ΔacrAB strain KAM3 conferred resistance to a range of drugs, but most significant resistance was to the macrolide spiramycin. A gel-shift analysis was used to determine if VarR bound to the promoter regions of the resistance genes. Consistent with the regulation of these resistance genes, VarR binds to three distinct intergenic regions, varRG, varGA and varBC located upstream and adjacent to varG, varA and varC, respectively. VarR can act as a repressor at the varRG promoter region; whilst this repression was relieved upon addition of β-lactams, these did not dissociate the VarR/varRG-DNA complex, indicating that the de-repression of varR by β-lactams is indirect. Considering that the genomic arrangement of VarR-VarG is strikingly similar to that of AmpR-AmpC system, it is possible that V. cholerae has evolved a system for resistance to the newer β-lactams that would prove more beneficial to the bacterium in light of current selective pressures.

  3. Yersinia pestis lineages in Mongolia.

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    Julia M Riehm

    Full Text Available BACKGROUND: Whole genome sequencing allowed the development of a number of high resolution sequence based typing tools for Yersinia (Y. pestis. The application of these methods on isolates from most known foci worldwide and in particular from China and the Former Soviet Union has dramatically improved our understanding of the population structure of this species. In the current view, Y. pestis including the non or moderate human pathogen Y. pestis subspecies microtus emerged from Yersinia pseudotuberculosis about 2,600 to 28,600 years ago in central Asia. The majority of central Asia natural foci have been investigated. However these investigations included only few strains from Mongolia. METHODOLOGY/PRINCIPAL FINDINGS: Clustered Regularly Interspaced Short Prokaryotic Repeats (CRISPR analysis and Multiple-locus variable number of tandem repeats (VNTR analysis (MLVA with 25 loci was performed on 100 Y. pestis strains, isolated from 37 sampling areas in Mongolia. The resulting data were compared with previously published data from more than 500 plague strains, 130 of which had also been previously genotyped by single nucleotide polymorphism (SNP analysis. The comparison revealed six main clusters including the three microtus biovars Ulegeica, Altaica, and Xilingolensis. The largest cluster comprises 78 isolates, with unique and new genotypes seen so far in Mongolia only. Typing of selected isolates by key SNPs was used to robustly assign the corresponding clusters to previously defined SNP branches. CONCLUSIONS/SIGNIFICANCE: We show that Mongolia hosts the most recent microtus clade (Ulegeica. Interestingly no representatives of the ancestral Y. pestis subspecies pestis nodes previously identified in North-western China were identified in this study. This observation suggests that the subsequent evolution steps within Y. pestis pestis did not occur in Mongolia. Rather, Mongolia was most likely re-colonized by more recent clades coming back from

  4. Rapid detection and strain typing of Chlamydia trachomatis using a highly multiplexed microfluidic PCR assay.

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    Rosemary S Turingan

    Full Text Available Nucleic acid amplification tests (NAATs are recommended by the CDC for detection of Chlamydia trachomatis (Ct urogenital infections. Current commercial NAATs require technical expertise and sophisticated laboratory infrastructure, are time-consuming and expensive, and do not differentiate the lymphogranuloma venereum (LGV strains that require a longer duration of treatment than non-LGV strains. The multiplexed microfluidic PCR-based assay presented in this work simultaneously interrogates 13 loci to detect Ct and identify LGV and non-LGV strain-types. Based on amplified fragment length polymorphisms, the assay differentiates LGV, ocular, urogenital, and proctocolitis clades, and also serovars L1, L2, and L3 within the LGV group. The assay was evaluated in a blinded fashion using 95 clinical swabs, with 76 previously reported as urogenital Ct-positive samples and typed by ompA genotyping and/or Multi-Locus Sequence Typing. Results of the 13-plex assay showed that 51 samples fell within urogenital clade 2 or 4, 24 samples showed both clade 2 and 4 signatures, indicating possible mixed infection, gene rearrangement, or inter-clade recombination, and one sample was a noninvasive trachoma biovar (either a clade 3 or 4. The remaining 19 blinded samples were correctly identified as LGV clade 1 (3, ocular clade 3 (4, or as negatives (12. To date, no NAAT assay can provide a point-of-care applicable turnaround time for Ct detection while identifying clinically significant Ct strain types to inform appropriate treatment. Coupled with rapid DNA processing of clinical swabs (approximately 60 minutes from swab-in to result-out, the assay has significant potential as a rapid POC diagnostic for Ct infections.

  5. High bacterial loads of Ureaplasma may be associated with non-specific cervicitis.

    Science.gov (United States)

    Liu, Lu; Cao, Guojun; Zhao, Zhen; Zhao, Fang; Huang, Yanqun

    2014-09-01

    Ureaplasma parvum and Ureaplasma urealyticum are commonly found in the cervix of women with non-chlamydial and non-gonococcal cervicitis or non-specific cervicitis (NSC). However their contribution to the aetiology of NSC is controversial. U. parvum and U. urealyticum were identified and quantified in cervical swabs collected from 155 women with NSC and 312 controls without NSC, using real-time PCR. The relative bacterial quantification was then calculated using the Ureaplasma copy number divided by the number of host cells; this is important for the correction of bias linked to the number of cells harvested in different swabs. Ureaplasma was detected in 58.7% (91/155) of NSC patients: U. parvum in 30.3%, U. urealyticum in 16.1%, and mixed infection in 12.3%. It was also detected in 54.5% (170/312) of controls: U. parvum in 33.0%, U. urealyticum in 11.5%, and mixed infection in 9.9%. There were no significant differences for U. parvum, U. urealyticum, or mixed infection between the 2 groups (p > 0.05). However, both biovars were present at higher concentrations in NSC patients than in controls (p 10 copies/1000 cells as a reference, the positive rate of U. parvum in NSC patients was 16.1%, significantly higher than that in controls at 5.1% (relative risk 3.145, p Ureaplasma can adhere to host cells, colonize, internalize, and subsequently produce pathological lesions. A high density of Ureaplasma in the cervix may be associated with the aetiology of NSC.

  6. Analysis of mutations in DNA gyrase and topoisomerase IV of Ureaplasma urealyticum and Ureaplasma parvum serovars resistant to fluoroquinolones.

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    Piccinelli, Giorgio; Gargiulo, Franco; Biscaro, Valeria; Caccuri, Francesca; Caruso, Arnaldo; De Francesco, Maria Antonia

    2017-01-01

    This study aims to determine the prevalence of fluoroquinolone resistance of Ureaplasma biovars and serovars isolated from urogenital clinical samples and determine the underlying molecular mechanism for quinolone resistance for all resistant isolates. Of 105 samples confirmed as positive for U. urealyticum/U. parvum, 85 were resistant to quinolones by the Mycoplasma-IST2 kit. However, only 43 out of 85 quinolone resistant isolates had amino acid substitutions in GyrA, GyrB, ParC and ParE proteins underlining that this assay have mis-identified as fluoroquinolone resistant 42 isolates. The known ParC E87K and ParC S83L mutations were found in 1 and 10 isolates, respectively. An original mutation of ureaplasmal ParC (E87Q, 1 isolate) was found. Furthermore, we found a ParE R448K mutation in one isolate, already described. Among the additional alterations detected, the most prevalent mutation found was L176F in GyrA protein in 18 isolates with single infection and in 3 isolates with mixed ureaplasma infections. Mutations in GyrB (E502Q, 4 isolates), ParE (Q412K, Q412P, Q412T, 3 independent isolates), whose role is unknown, were also found. Other sporadic mutations in the four genes were identified. This investigation is the result of monitoring the data for molecular fluoroquinone resistance in Ureaplasma spp. in Italy. Resulting that this acquired resistance is high and that continued local epidemiological studies are essential to monitor and document their antimicrobial resistance trends. Copyright © 2016 Elsevier B.V. All rights reserved.

  7. Re-evaluation of the taxonomy of the Mitis group of the genus Streptococcus based on whole genome phylogenetic analyses, and proposed reclassification of Streptococcus dentisani as Streptococcus oralis subsp. dentisani comb. nov., Streptococcus tigurinus as Streptococcus oralis subsp. tigurinus comb. nov., and Streptococcus oligofermentans as a later synonym of Streptococcus cristatus.

    Science.gov (United States)

    Jensen, Anders; Scholz, Christian F P; Kilian, Mogens

    2016-11-01

    The Mitis group of the genus Streptococcus currently comprises 20 species with validly published names, including the pathogen S. pneumoniae. They have been the subject of much taxonomic confusion, due to phenotypic overlap and genetic heterogeneity, which has hampered a full appreciation of their clinical significance. The purpose of this study was to critically re-examine the taxonomy of the Mitis group using 195 publicly available genomes, including designated type strains for phylogenetic analyses based on core genomes, multilocus sequences and 16S rRNA gene sequences, combined with estimates of average nucleotide identity (ANI) and in silico and in vitro analyses of specific phenotypic characteristics. Our core genomic phylogenetic analyses revealed distinct clades that, to some extent, and from the clustering of type strains represent known species. However, many of the genomes have been incorrectly identified adding to the current confusion. Furthermore, our data show that 16S rRNA gene sequences and ANI are unsuitable for identifying and circumscribing new species of the Mitis group of the genus Streptococci. Based on the clustering patterns resulting from core genome phylogenetic analysis, we conclude that S. oligofermentans is a later synonym of S. cristatus. The recently described strains of the species Streptococcus dentisani includes one previously referred to as 'S. mitis biovar 2'. Together with S. oralis, S. dentisani and S. tigurinus form subclusters within a coherent phylogenetic clade. We propose that the species S. oralis consists of three subspecies: S. oralis subsp. oralis subsp. nov., S. oralis subsp. tigurinus comb. nov., and S. oralis subsp. dentisani comb. nov.

  8. A Bacterial Glycoengineered Antigen for Improved Serodiagnosis of Porcine Brucellosis.

    Science.gov (United States)

    Cortina, María E; Balzano, Rodrigo E; Rey Serantes, Diego A; Caillava, Ana J; Elena, Sebastián; Ferreira, A C; Nicola, Ana M; Ugalde, Juan E; Comerci, Diego J; Ciocchini, Andrés E

    2016-06-01

    Brucellosis is a highly zoonotic disease that affects animals and human beings. Brucella suis is the etiological agent of porcine brucellosis and one of the major human brucellosis pathogens. Laboratory diagnosis of porcine brucellosis mainly relies on serological tests, and it has been widely demonstrated that serological assays based on the detection of anti O-polysaccharide antibodies are the most sensitive tests. Here, we validate a recombinant glycoprotein antigen, an N-formylperosamine O-polysaccharide-protein conjugate (OAg-AcrA), for diagnosis of porcine brucellosis. An indirect immunoassay based on the detection of anti-O-polysaccharide IgG antibodies was developed coupling OAg-AcrA to enzyme-linked immunosorbent assay plates (glyco-iELISA). To validate the assay, 563 serum samples obtained from experimentally infected and immunized pigs, as well as animals naturally infected with B. suis biovar 1 or 2, were tested. A receiver operating characteristic (ROC) analysis was performed, and based on this analysis, the optimum cutoff value was 0.56 (relative reactivity), which resulted in a diagnostic sensitivity and specificity of 100% and 99.7%, respectively. A cutoff value of 0.78 resulted in a test sensitivity of 98.4% and a test specificity of 100%. Overall, our results demonstrate that the glyco-iELISA is highly accurate for diagnosis of porcine brucellosis, improving the diagnostic performance of current serological tests. The recombinant glycoprotein OAg-AcrA can be produced in large homogeneous batches in a standardized way, making it an ideal candidate for further validation as a universal antigen for diagnosis of "smooth" brucellosis in animals and humans. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  9. Cellulose Synthesis in Agrobacterium tumefaciens

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    Alan R. White; Ann G. Matthysse

    2004-07-31

    We have cloned the celC gene and its homologue from E. coli, yhjM, in an expression vector and expressed the both genes in E. coli; we have determined that the YhjM protein is able to complement in vitro cellulose synthesis by extracts of A. tumefaciens celC mutants, we have purified the YhjM protein product and are currently examining its enzymatic activity; we have examined whole cell extracts of CelC and various other cellulose mutants and wild type bacteria for the presence of cellulose oligomers and cellulose; we have examined the ability of extracts of wild type and cellulose mutants including CelC to incorporate UDP-14C-glucose into cellulose and into water-soluble, ethanol-insoluble oligosaccharides; we have made mutants which synthesize greater amounts of cellulose than the wild type; and we have examined the role of cellulose in the formation of biofilms by A. tumefaciens. In addition we have examined the ability of a putative cellulose synthase gene from the tunicate Ciona savignyi to complement an A. tumefaciens celA mutant. The greatest difference between our knowledge of bacterial cellulose synthesis when we started this project and current knowledge is that in 1999 when we wrote the original grant very few bacteria were known to synthesize cellulose and genes involved in this synthesis were sequenced only from Acetobacter species, A. tumefaciens and Rhizobium leguminosarum. Currently many bacteria are known to synthesize cellulose and genes that may be involved have been sequenced from more than 10 species of bacteria. This additional information has raised the possibility of attempting to use genes from one bacterium to complement mutants in another bacterium. This will enable us to examine the question of which genes are responsible for the three dimensional structure of cellulose (since this differs among bacterial species) and also to examine the interactions between the various proteins required for cellulose synthesis. We have carried out one

  10. Solubilisation of inorganic phosphates by inoculant strains from tropical legumes

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    Leandro Marciano Marra

    2011-10-01

    Full Text Available Microbial solubilisation of low soluble inorganic phosphates is an important process contributing for the phosphorus available to plants in tropical soils. This study evaluates the ability of inoculant strains for tropical legumes to solubilise inorganic phosphates of low solubility that are found in tropical soils. Seven strains of Leguminosae nodulating bacteria (LNB were compared with one another and with a non-nodulating positive control, Burkholderia cepacia (LMG 1222T. Four of the strains are used as inoculants for cowpeas (Vigna unguiculata (Bradyrhizobium sp. UFLA 03-84; Bradyrhizobium elkani INPA 03-11B and Bradyrhizobium japonicum BR3267 or for common beans (Phaseolus vulgaris (Rhizobium tropici CIAT 899T. Rhizobium etli UFLA 02-100 and Rhizobium leguminosarum 316C10a are also efficient nodulators of beans and Cupriavidus taiwanensis LMG 19424T nodulates on Mimosa pudica. Two experiments, with solid and liquid media, were performed to determine whether the strains were able to solubilise CaHPO4, Al(H2PO43 or FePO4.2H2O. On solid GELP medium none of the strains dissolved FePO4.2H2O, but LMG 1222, UFLA 03-84 and CIAT 899 solubilised CaHPO4 particularly well. These strains, along with LMG 19424 and BR 3267, were also able to increase the solubility of Al(H2PO43. In liquid GELP medium, LMG 1222 solubilised all phosphate sources, but no legume nodulating strain could increase the solubility of Al(H2PO43. The strains CIAT 899 and UFLA 02-100 were the only legume nodulating bacteria able to solubilise the other phosphate sources in liquid media, dissolving both CaHPO4 and FePO4.2H2O. There was a negative correlation between the pH of the culture medium and the concentration of soluble phosphate when the phosphorus source was CaHPO4 or FePO4.2H2O. The contribution of these strains to increasing the phosphorus nutrition of legumes and non-legume plant species should be investigated further by in vivo experiments.

  11. Descrição, produtividade e estabilidade da cultivar de soja IAC-23, resistente a insetos Description, yield and stability of early soybean insect resistant cultivar IAC-23

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    Manoel Albino Coelho de Miranda

    2003-01-01

    Ribeirão Preto (1996/97; and Campinas (1998/99. Conclusive trials were conducted in randomized blocks with four replications. This cultivar shows long juveniIe period, flowering time at 43 days, and complete life cicle around 106 days after seed germination, with plant height about 67 cm. The average yield was 3017 kg.ha-1. Plants show brown pubescence, yellow seeds with brown hilum. The new cultivar is resistant to: bacterial pustule (Xanthomonas campestris pv. glycines, wildfire (Pseudomonas syringae pv. tabaci, stem canker (Phomopsis phaseoli f.sp. meridionalis and soybean mosaic birus (SMV. IAC-23 productivity, large stability and resistance levels to leaf and pod feeders insects is similar to that observed for IAC-17, and superior to that of IAS-5. The cultivar IAC-23 is then recommended for cultivation in the State of São Paulo and in similar environmental conditions.

  12. Efficacy of standard therapies against Ureaplasma species and persistence among men with non-gonococcal urethritis enrolled in a randomised controlled trial.

    Science.gov (United States)

    Khosropour, Christine M; Manhart, Lisa E; Gillespie, Catherine W; Lowens, M Sylvan; Golden, Matthew R; Jensen, Nicole L; Kenny, George E; Totten, Patricia A

    2015-08-01

    Ureaplasma urealyticum biovar 2 (UU-2), but not Ureaplasma parvum (UP), has been associated with non-gonococcal urethritis (NGU), but little is known about species-specific responses to standard therapies. We examined species-specific treatment outcomes and followed men with treatment failure for 9 weeks. From May 2007 to July 2011, men aged ≥16 attending a sexually transmitted disease (STD) clinic in Seattle, Washington, with NGU (urethral discharge or urethral symptoms plus ≥5 polymorphonuclear leucocytes /high-powered field) enrolled in a double-blind, randomised trial. Participants received active azithromycin (1 g) + placebo doxycycline or active doxycycline (100 mg twice a day ×7 days) + placebo azithromycin. Ureaplasma were detected in culture followed by species-specific PCR. Outcomes were assessed at 3, 6 and 9 weeks. At 3 weeks, men with persistent Ureaplasma detection received 'reverse therapy' (e.g., active doxycycline if they first received active azithromycin). At 6 weeks, persistently positive men received moxifloxacin (400 mg×7 days). Of 490 men, 107 (22%) and 60 (12%) were infected with UU-2 and UP, respectively, and returned at 3 weeks. Persistent detection was similar for UU-2-infected men initially treated with azithromycin or doxycycline (25% vs. 31%; p=0.53), but differed somewhat for men with UP (45% vs. 24%; p=0.11). At 6 weeks, 57% of UU-2-infected and 63% of UP-infected men who received both drugs had persistent detection. Failure after moxifloxacin occurred in 30% and 36%, respectively. Persistent detection of UU-2 or UP was not associated with signs/symptoms of NGU. Persistent detection after treatment with doxycycline, azithromycin and moxifloxacin was common for UU and UP, but not associated with persistent urethritis. NCT00358462. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  13. Bacterial Vaginosis–Associated Bacteria in Men: Association of Leptotrichia/Sneathia spp. With Nongonococcal Urethritis

    Science.gov (United States)

    Manhart, Lisa E.; Khosropour, Christine M.; Liu, Congzhu; Gillespie, Catherine W.; Depner, Kevin; Fiedler, Tina; Marrazzo, Jeanne M.; Fredricks, David N.

    2014-01-01

    Background Approximately 45% of nongonococcal urethritis cases have no identified etiology. Novel bacteria recently associated with bacterial vaginosis (BV) in women may be involved. We evaluated the association of idiopathic nongonococcal urethritis and 5 newly described BV-associated bacteria (BVAB). Methods Heterosexual men 16 years or older attending a sexually transmitted disease clinic in Seattle, Washington, from May 2007 to July 2011 and negative for Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Mycoplasma genitalium, and Ureaplasma urealyticum–biovar2 were eligible. Cases had urethral discharge or 5 or more polymorphonuclear leukocytes per high-power field in urethral exudates. Controls had no urethral discharge and less than 5 polymorphonuclear leukocytes per high-power field. Urine was tested for Atopobium spp., BVAB-2, BVAB-3, Megasphaera spp., and Leptotrichia/Sneathia spp. using quantitative taxon-directed polymerase chain reaction. Results Cases (n = 157) and controls (n = 102) were of similar age, education, and income, and most were white. Leptotrichia/Sneathia spp. was significantly associated with urethritis (24/157 [15.3%] vs. 6/102 [5.9%], P = 0.03). BVAB-2 was more common in cases than in controls (7/157 [4.5%] vs. 1/102 [1.0%], P = 0.15), and BVAB-3 (n = 2) and Megasphaera spp. (n = 1) were only detected in men with urethritis, but these bacteria were found only in men who also had Leptotrichia/Sneathia spp. Atopobium spp. was not associated with urethritis. The quantity of bacteria did not differ between cases and controls. Among treated cases, doxycycline was more effective than azithromycin for clinical cure of men with Leptotrichia/Sneathia spp. (9/10 vs. 7/12, P = 0.16) and BVAB-2 (3/3 vs. 0/3, P = 0.10). Conclusions Leptotrichia/Sneathia spp. may be urethral pathogens or contribute to a pathogenic microbiota that can also include BVAB-2, BVAB-3, and Megasphaera spp. Doxycycline may be more effective than

  14. [Plague in Zaire].

    Science.gov (United States)

    Janssens, P G; Pattyn, S R

    1994-01-01

    Two endemic foci of plague have been discovered in Zaïre, the first in the Ituri in 1928, the other in North-Kivu in 1938. They are situated in the region of the great East-African Rift and are adjacent to the Ugandan focus, identified in 1877. A strict surveillance of these endemic foci makes it possible to state that, between 1928 and 1959, 632 cases of plague have been diagnosed in the Ituri, or 20 a year, and 190 in the N-Kivu, or 8 a year. Since then several flare ups have been notified. This situation is very remote from the "black death" concept. Yersinia pestis presents, besides its bipolar staining, many other characteristics such as the indispensable presence of iron to produce virulence, or the fermentation of glycerine and reduction of nitrates as parameters for the identification of 3 biovars, corresponding with a specific geographic distribution: antiqua, medievalis, orientalis or maritima. The antigenic structure has been discussed and also the role of plasmids. Plague is a disease of rats, a variegated gathering of rodents with different degrees of tolerance and sensitiveness to Y.pestis, living in a frail equilibrium. The multimammate houserat was in the Ituri the principal agent until the black rat Rattus rattus invaded the region and a new balance came into being. The frequent changes in taxonomy of Mastomys caused uncertainties. The transmission is due to fleas subject to a blocking of their ventriculum by Y.pestis. Fleas play an active part in the process. Man is only a casual intruder. The pathogenicity is related to its invasiveness and its intracellular localization in macrophages and other R.E. cells, in which Y.pestis can survive. The bubo is characteristic of the disease. In Zaïre a septicaemic tendency has been observed, with a possible involvement of the C.N.S. and of the lungs. The latter may produce among the surrounding relatives primary pneumonic plague. The clinical diagnosis ought to be confirmed by bacteriologic investigation

  15. Bacterial vaginosis-associated bacteria in men: association of Leptotrichia/Sneathia spp. with nongonococcal urethritis.

    Science.gov (United States)

    Manhart, Lisa E; Khosropour, Christine M; Liu, Congzhu; Gillespie, Catherine W; Depner, Kevin; Fiedler, Tina; Marrazzo, Jeanne M; Fredricks, David N

    2013-12-01

    Approximately 45% of nongonococcal urethritis cases have no identified etiology. Novel bacteria recently associated with bacterial vaginosis (BV) in women may be involved. We evaluated the association of idiopathic nongonococcal urethritis and 5 newly described BV-associated bacteria (BVAB). Heterosexual men 16 years or older attending a sexually transmitted disease clinic in Seattle, Washington, from May 2007 to July 2011 and negative for Chlamydia trachomatis, Neisseria gonorrhoeae, Trichomonas vaginalis, Mycoplasma genitalium, and Ureaplasma urealyticum-biovar2 were eligible. Cases had urethral discharge or 5 or more polymorphonuclear leukocytes per high-power field in urethral exudates. Controls had no urethral discharge and less than 5 polymorphonuclear leukocytes per high-power field. Urine was tested for Atopobium spp., BVAB-2, BVAB-3, Megasphaera spp., and Leptotrichia/Sneathia spp. using quantitative taxon-directed polymerase chain reaction. Cases (n = 157) and controls (n = 102) were of similar age, education, and income, and most were white. Leptotrichia/Sneathia spp. was significantly associated with urethritis (24/157 [15.3%] vs. 6/102 [5.9%], P = 0.03). BVAB-2 was more common in cases than in controls (7/157 [4.5%] vs. 1/102 [1.0%], P = 0.15), and BVAB-3 (n = 2) and Megasphaera spp. (n = 1) were only detected in men with urethritis, but these bacteria were found only in men who also had Leptotrichia/Sneathia spp. Atopobium spp. was not associated with urethritis. The quantity of bacteria did not differ between cases and controls. Among treated cases, doxycycline was more effective than azithromycin for clinical cure of men with Leptotrichia/Sneathia spp. (9/10 vs. 7/12, P = 0.16) and BVAB-2 (3/3 vs. 0/3, P = 0.10). Leptotrichia/Sneathia spp. may be urethral pathogens or contribute to a pathogenic microbiota that can also include BVAB-2, BVAB-3, and Megasphaera spp. Doxycycline may be more effective than azithromycin against these newly identified

  16. Metagenomic Analysis of Dairy Bacteriophages: Extraction Method and Pilot Study on Whey Samples Derived from Using Undefined and Defined Mesophilic Starter Cultures.

    Science.gov (United States)

    Muhammed, Musemma K; Kot, Witold; Neve, Horst; Mahony, Jennifer; Castro-Mejía, Josué L; Krych, Lukasz; Hansen, Lars H; Nielsen, Dennis S; Sørensen, Søren J; Heller, Knut J; van Sinderen, Douwe; Vogensen, Finn K

    2017-10-01

    Despite being potentially highly useful for characterizing the biodiversity of phages, metagenomic studies are currently not available for dairy bacteriophages, partly due to the lack of a standard procedure for phage extraction. We optimized an extraction method that allows the removal of the bulk protein from whey and milk samples with losses of less than 50% of spiked phages. The protocol was applied to extract phages from whey in order to test the notion that members of Lactococcus lactis 936 (now Sk1virus ), P335, c2 (now C2virus ) and Leuconostoc phage groups are the most frequently encountered in the dairy environment. The relative abundance and diversity of phages in eight and four whey mixtures from dairies using undefined mesophilic mixed-strain cultures containing Lactococcus lactis subsp. lactis biovar diacetylactis and Leuconostoc species (i.e., DL starter cultures) and defined cultures, respectively, were assessed. Results obtained from transmission electron microscopy and high-throughput sequence analyses revealed the dominance of Lc. lactis 936 phages (order Caudovirales , family Siphoviridae ) in dairies using undefined DL starter cultures and Lc. lactis c2 phages (order Caudovirales , family Siphoviridae ) in dairies using defined cultures. The 936 and Leuconostoc phages demonstrated limited diversity. Possible coinduction of temperate P335 prophages and satellite phages in one of the whey mixtures was also observed. IMPORTANCE The method optimized in this study could provide an important basis for understanding the dynamics of the phage community (abundance, development, diversity, evolution, etc.) in dairies with different sizes, locations, and production strategies. It may also enable the discovery of previously unknown phages, which is crucial for the development of rapid molecular biology-based methods for phage burden surveillance systems. The dominance of only a few phage groups in the dairy environment signifies the depth of knowledge

  17. Brucella papionis sp. nov., isolated from baboons (Papio spp.).

    Science.gov (United States)

    Whatmore, Adrian M; Davison, Nicholas; Cloeckaert, Axel; Al Dahouk, Sascha; Zygmunt, Michel S; Brew, Simon D; Perrett, Lorraine L; Koylass, Mark S; Vergnaud, Gilles; Quance, Christine; Scholz, Holger C; Dick, Edward J; Hubbard, Gene; Schlabritz-Loutsevitch, Natalia E

    2014-12-01

    Two Gram-negative, non-motile, non-spore-forming coccoid bacteria (strains F8/08-60(T) and F8/08-61) isolated from clinical specimens obtained from baboons (Papio spp.) that had delivered stillborn offspring were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA gene sequence similarities, both strains, which possessed identical sequences, were assigned to the genus Brucella. This placement was confirmed by extended multilocus sequence analysis (MLSA), where both strains possessed identical sequences, and whole-genome sequencing of a representative isolate. All of the above analyses suggested that the two strains represent a novel lineage within the genus Brucella. The strains also possessed a unique profile when subjected to the phenotyping approach classically used to separate species of the genus Brucella, reacting only with Brucella A monospecific antiserum, being sensitive to the dyes thionin and fuchsin, being lysed by bacteriophage Wb, Bk2 and Fi phage at routine test dilution (RTD) but only partially sensitive to bacteriophage Tb, and with no requirement for CO2 and no production of H2S but strong urease activity. Biochemical profiling revealed a pattern of enzyme activity and metabolic capabilities distinct from existing species of the genus Brucella. Molecular analysis of the omp2 locus genes showed that both strains had a novel combination of two highly similar omp2b gene copies. The two strains shared a unique fingerprint profile of the multiple-copy Brucella-specific element IS711. Like MLSA, a multilocus variable number of tandem repeat analysis (MLVA) showed that the isolates clustered together very closely, but represent a distinct group within the genus Brucella. Isolates F8/08-60(T) and F8/08-61 could be distinguished clearly from all known species of the genus Brucella and their biovars by both phenotypic and molecular properties. Therefore, by applying the species concept for the genus Brucella suggested by the ICSP

  18. Brucella microti sp. nov., isolated from the common vole Microtus arvalis.

    Science.gov (United States)

    Scholz, Holger C; Hubalek, Zdenek; Sedlácek, Ivo; Vergnaud, Gilles; Tomaso, Herbert; Al Dahouk, Sascha; Melzer, Falk; Kämpfer, Peter; Neubauer, Heinrich; Cloeckaert, Axel; Maquart, Marianne; Zygmunt, Michel S; Whatmore, Adrian M; Falsen, Enevold; Bahn, Peter; Göllner, Cornelia; Pfeffer, Martin; Huber, Birgit; Busse, Hans-Jürgen; Nöckler, Karsten

    2008-02-01

    Two Gram-negative, non-motile, non-spore-forming, coccoid bacteria (strains CCM 4915(T) and CCM 4916), isolated from clinical specimens of the common vole Microtus arvalis during an epizootic in the Czech Republic in 2001, were subjected to a polyphasic taxonomic study. On the basis of 16S rRNA (rrs) and recA gene sequence similarities, both isolates were allocated to the genus Brucella. Affiliation to Brucella was confirmed by DNA-DNA hybridization studies. Both strains reacted equally with Brucella M-monospecific antiserum and were lysed by the bacteriophages Tb, Wb, F1 and F25. Biochemical profiling revealed a high degree of enzyme activity and metabolic capabilities not observed in other Brucella species. The omp2a and omp2b genes of isolates CCM 4915(T) and CCM 4916 were indistinguishable. Whereas omp2a was identical to omp2a of brucellae from certain pinniped marine mammals, omp2b clustered with omp2b of terrestrial brucellae. Analysis of the bp26 gene downstream region identified strains CCM 4915(T) and CCM 4916 as Brucella of terrestrial origin. Both strains harboured five to six copies of the insertion element IS711, displaying a unique banding pattern as determined by Southern blotting. In comparative multilocus VNTR (variable-number tandem-repeat) analysis (MLVA) with 296 different genotypes, the two isolates grouped together, but formed a separate cluster within the genus Brucella. Multilocus sequence typing (MLST) analysis using nine different loci also placed the two isolates separately from other brucellae. In the IS711-based AMOS PCR, a 1900 bp fragment was generated with the Brucella ovis-specific primers, revealing that the insertion element had integrated between a putative membrane protein and cboL, encoding a methyltransferase, an integration site not observed in other brucellae. Isolates CCM 4915(T) and CCM 4916 could be clearly distinguished from all known Brucella species and their biovars by means of both their phenotypic and molecular

  19. Adaptation of Microstix®-Candida Slide-test for Diagnosis of Bovine Mastitis Due to Anascogenic Yeasts

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    Przemysław Dudko

    2010-01-01

    Full Text Available Although a large diversity of mastitis-causing bacteria (150 species, subspecies or biovars have been documented, an adequate and simple mycological screening for yeast-related mastitis has not been developed. Since yeast incubation normally lasts longer than the majority of bacteriological ones, usually a treatment is administered before the results of microbial cultures are obtained. Therefore, a simple test has been needed for detection of yeast-related mastitis, the results of which can be read in parallel to bacteriological screening. Application of Microstix®-Candida assay was first checked for its specificity in standard Candida albicans (Oxford strain cultures (phase I and then in the other 36 strains of yeasts isolated from clinical mastitis cases during field investigations (second phase. Next, a test was implemented for identification of pathogens in 9 cohorts (1,200 cows from red-white Polish- Friesian breed in which the dynamics of mastitis were traced by bacteriological and mycological assays along year-long observations. The data obtained showed that apart from frequent infections (~50-70% proved by using pure microbial cultures, also mixed (bacterial-fungal infections (0-25% appeared as well as negative sieves (in the range of 0-45%. Mycological assays revealed a significant fraction of mixed infections during both lactation and the dry period. Strepto-staphylococcal infection was thus identified, however, its proportion strongly decreased after the introduction of antibiotics (after sensitivity to microbial agents was determined. Microbial monitoring was improved in these herds by parallel application of bacteriological and mycological (particularly Microstix®- Candida tests. Microstix®-assay appears to be useful for mycological diagnosis in field conditions of infected (mastitis herds. Its advantages include easy performance and short incubation time that is only 24 h at 37 °C (72-96 h at room temperature. This allowed

  20. Characterization of Zur-dependent genes and direct Zur targets in Yersinia pestis

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    Wang Xiaoyi

    2009-06-01

    Full Text Available Abstract Background The zinc uptake regulator Zur is a Zn2+-sensing metalloregulatory protein involved in the maintenance of bacterial zinc homeostasis. Up to now, regulation of zinc homeostasis by Zur is poorly understood in Y. pestis. Results We constructed a zur null mutant of Y. pestis biovar microtus strain 201. Microarray expression analysis disclosed a set of 154 Zur-dependent genes of Y. pestis upon exposure to zinc rich condition. Real-time reverse transcription (RT-PCR was subsequently used to validate the microarray data. Based on the 154 Zur-dependent genes, predicted regulatory Zur motifs were used to screen for potential direct Zur targets including three putative operons znuA, znuCB and ykgM-RpmJ2. The LacZ reporter fusion analysis verified that Zur greatly repressed the promoter activity of the above three operons. The subsequent electrophoretic mobility shift assay (EMSA demonstrated that a purified Zur protein was able to bind to the promoter regions of the above three operons. The DNase I footprinting was used to identify the Zur binding sites for the above three operons, verifying the Zur box sequence as predicted previously in γ-Proteobacteria. The primer extension assay was further used to determine the transcription start sites for the above three operons and to localize the -10 and -35 elements. Zur binding sites overlapped the -10 sequence of its target promoters, which was consistent with the previous observation that Zur binding would block the entry of the RNA polymerase to repress the transcription of its target genes. Conclusion Zur as a repressor directly controls the transcription of znuA, znuCB and ykgM-RpmJ2 in Y. pestis by employing a conserved mechanism of Zur-promoter DNA association as observed in γ-Proteobacteria. Zur contributes to zinc homeostasis in Y. pestis likely through transcriptional repression of the high-affinity zinc uptake system ZnuACB and two alternative ribosomal proteins YkgM and RpmJ2.

  1. MLVA-16 typing of 295 marine mammal Brucella isolates from different animal and geographic origins identifies 7 major groups within Brucella ceti and Brucella pinnipedialis

    Directory of Open Access Journals (Sweden)

    Jacques Isabelle

    2009-07-01

    Full Text Available Abstract Background Since 1994, Brucella strains have been isolated from a wide range of marine mammals. They are currently recognized as two new Brucella species, B. pinnipedialis for the pinniped isolates and B. ceti for the cetacean isolates in agreement with host preference and specific phenotypic and molecular markers. In order to investigate the genetic relationships within the marine mammal Brucella isolates and with reference to terrestrial mammal Brucella isolates, we applied in this study the Multiple Loci VNTR (Variable Number of Tandem Repeats Analysis (MLVA approach. A previously published assay comprising 16 loci (MLVA-16 that has been shown to be highly relevant and efficient for typing and clustering Brucella strains from animal and human origin was used. Results 294 marine mammal Brucella strains collected in European waters from 173 animals and a human isolate from New Zealand presumably from marine origin were investigated by MLVA-16. Marine mammal Brucella isolates were shown to be different from the recognized terrestrial mammal Brucella species and biovars and corresponded to 3 major related groups, one specific of the B. ceti strains, one of the B. pinnipedialis strains and the last composed of the human isolate. In the B. ceti group, 3 subclusters were identified, distinguishing a cluster of dolphin, minke whale and porpoise isolates and two clusters mostly composed of dolphin isolates. These results were in accordance with published analyses using other phenotypic or molecular approaches, or different panels of VNTR loci. The B. pinnipedialis group could be similarly subdivided in 3 subclusters, one composed exclusively of isolates from hooded seals (Cystophora cristata and the two others comprising other seal species isolates. Conclusion The clustering analysis of a large collection of marine mammal Brucella isolates from European waters significantly strengthens the current view of the population structure of these two

  2. Eficiência e competitividade de variantes espontâneos isolados de estirpes de Bradyrhizobium spp recomendadas para a cultura da soja (Glycine max Effectiveness and competitiveness of spontaneous mutants isolated from Bradyrhizobium spp strains recommended for soybean crop (Glycine max

    Directory of Open Access Journals (Sweden)

    Fabíola Gomes de Carvalho

    2005-12-01

    Full Text Available O cultivo sucessivo de soja inoculada numa mesma área proporcionou a adaptação de uma população de rizóbios, que podem não ser tão eficientes quanto à capacidade de fixação de N2, mas apresentam alta competitividade, dificultando a introdução de novas estirpes mais eficientes. Com a finalidade de avaliar o desempenho simbiótico (eficiência e competitividade de variantes espontâneos isolados de estirpes de B. japonicum (SEMIA 5079 e SEMIA 5080 e B. elkanii (SEMIA 587 e SEMIA 5019, realizou-se um experimento em casa de vegetação onde os variantes foram inoculados isoladamente e em diferentes combinações entre os variantes e uma estirpe comprovadamente mais competitiva (SEMIA 587 ou SEMIA 5019 a partir da adição de inóculos mistos (1/1; v/v no cultivar de soja BR-16. Por meio da avaliação das variáveis analisadas (nodulação, produção de matéria de seca da parte aérea, N total acumulado na parte aérea e ocupação nodular, foi possível constatar que o determinante da maior eficiência em tratamentos co-inoculados não foi a ocupação nodular de determinada estirpe ou variante presente no inóculo, mas, sim, o tipo de interação (sinérgica ou antagônica predominante no tratamento co-inoculado e que é possível selecionar variantes eficientes e competitivos para a cultura da soja a partir de estirpes parentais que já apresentam características desejáveis para utilização em inoculantes comerciais.The continuous cultivation of inoculated soybean in the same area can determine the soil colonization with a rhizobia population presenting low nitrogen fixation effectiveness. This fact can be a problem for the establishment of a more effective population. A greenhouse experiment was carried out to evaluate the symbiotic effectiveness and competitiveness of spontaneous mutants isolated from B. japonicum (SEMIA 5079 and SEMIA 5080 and B. elkanii (SEMIA 587 and SEMIA 5019 strains. The soybean biovar BR 16 was

  3. Epidemiological profile of reproductive loss in dairy cattle

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    Raul Costa Mascarenhas Santana

    2012-12-01

    -reactive with these titles. Between animals positive to the test, the lowest title observed was 1:25, while the highest obtained was 1:400. Viral neutralization tests demonstrated that 26.8% of the animals had titers greater or equal to 1:256, and among the animals with a history of abortions or stillbirths, 38.1% and 55.56% were serum reactive in the range considered, respectively. Between positive animals for serum neutralization technique, the lower titers observed were 1:2, while the highest obtained was equal to or greater than 1:1024. Only 5.56% of the animals studied had titers equal to or greater than 1:200 in the microscopic agglutination test for diagnosis of Leptospira spp. Only a single animal with a history of abortion did not presented negative serology (title 1:100. No animal with a history of stillbirth was presented in the reagent test. The prevalent biovar in reactive animals was Pomona (40%, Hardjo (30%, Tarassovi (20% and Wollfi (10%. There weren’t positive reactions to the tamponated acidified antigen test for diagnosis of bovine brucellosis. The study suggests the possibility of concurrent infections by other agents causing reproductive failure in dairy cattle.

  4. Molecular and phylogenetic analyses of Salmonella Gallinarum trace the origin and diversification of recent outbreaks of fowl typhoid in poultry farms.

    Science.gov (United States)

    De Carli, Silvia; Gräf, Tiago; Kipper, Diéssy; Lehmann, Fernanda Kieling Moreira; Zanetti, Nathalie; Siqueira, Franciele Maboni; Cibulski, Samuel; Fonseca, André Salvador Kazantzi; Ikuta, Nilo; Lunge, Vagner Ricardo

    2017-12-01

    Fowl typhoid (FT) and pullorum disease (PD) are two important poultry infections caused by Salmonella enterica subsp. enterica serotype Gallinarum (S. Gallinarum). S. Gallinarum strains are adapted to birds and classified into biovars Gallinarum (bvGA) and Pullorum (bvPU) as they are the causative agent of FT and PD, respectively. In Brazil, FT/PD outbreaks have been reported along the last 50 years, but there was a recent increase of FT field reports with the suspicion it could be due to virulence reversion of the attenuated live vaccine SG9R. In this study, we applied molecular biology assays and phylogenetic methods to detect and investigate S. Gallinarum isolates from commercial poultry flocks in order to understand the evolutionary history and origin of the recent FT outbreaks in Brazil. S. Gallinarum isolates were obtained from thirteen different poultry flocks with clinical signs of FT/PD from 2013 to 2015. These isolates were serotyped, tested with three specific PCR (for the detection of bvGA, bvPU and live vaccine strain SG9R) and submitted to sequencing of a variable genome region (ISR analysis). The complete genome of one bvGA strain (BR_RS12) was also compared to other S. Gallinarum complete genomes (including other two Brazilian ones: bvGA 287/91 and bvPU FCVA198). PCR detected all thirteen isolates as S. Gallinarum (eight bvGA and five bvPU), none positive for SG9R strain. ISR analysis revealed that all eight bvGA isolates showed exactly the same nucleotide sequences with 100% similarity to reference strains, while two patterns were observed for bvPU. Genome phylogeny demonstrated distinct clades for bvGA and bvPU, with the bvGA clade showing a clear subdivision including three genomes: SG9R vaccine, the respective SG9 parent strain and one SG9R revertant field isolate (MB4523). The evolutionary rate of the total S. Gallinarum genome was calculated at 6.15×10 -7 substitutions/site/year, with 2.8 observed substitutions per year per genome (1 SNP per

  5. Effect of simultaneous application of mycorrhiza with compost, vermicompost and sulfural geranole on some quantitative and qualitative characteristics of sesame (Sesamum indicum L. in a low input cropping system

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    P rezvani moghaddam

    2016-03-01

    quantitative and qualitative characteristics of plant, moreover, decreased environment risks of chemical inputs and ensure sustainability of production in long time by this approach. Acknowledgments We wish to thank Vice President for Research and Technology, Ferdowsi University of Mashhad, Iran for the financial support of the project. Keywords: Ecological inputs, Glomus mosseae, Health of food, Medicinal crops References Hawkes, C.V., Hartley, I.P., Ineson, P., and Fitter, A.H. 2008. Soil temperature affects carbon allocation within arbuscular mycorrhizal networks and carbon transport from plant to fungus. Global Change Biology 14: 1181-1190. Motta, S.R., and Maggiore, T. 2013. Evaluation of nitrogen management in maize cultivation grown on soil amended with sewage sludge and urea. European Journal of Agronomy 45: 59-67. Raja Sekar, K., and Karmegan, N. 2010. Earthworm casts as an alternate carrier material for biofertilizers: Assessment of endurance and viability of Azotobacter chroococcum, Bacillus megaterium and Rhizobium leguminosarum. Scientia Horticulturae 124: 286-289.

  6. 块根块茎类作物内生固氮菌分离鉴定、系统发育与促生特性%Isolation, Identification, Phylogeny and Growth Promoting Characteristics of Endophytic Diazotrophs from Tuber and Root Crops

    Institute of Scientific and Technical Information of China (English)

    李艳星; 郭平毅; 孙建光

    2017-01-01

    . phosphorivorans, Flavimonas oryzihabitans, Flavobacterium oncorhynchi, Microbacterium hydrocarbonoxydans, Microbacterium phyllosphaerae, Micrococcus endophyticus, Paenibacillus barengoltzii, Pae. cineris, Pae. glycanilyticus, Pae. lautus, Pae. tundrae, Pantoea agglomerans, Pan. anthophila, Pan. dispersa, Pan. rodasii, Pseudomonas azotoformans, Ps. beteli, Ps. brassicacearum, Ps. geniculata, Ps. hunanensis, Ps. koreensis, Ps. parafulva, Ps. seleniipraecipitans, Ps. simiae, Ps. syringae, Rahnella aquatilis, Rhizobium leguminosarum, Rh. massiliae, Rh. radiobacter, Sphingobacterium canadense, Sp. faecium, Staphylococcus sciuri, Stenotrophomonas rhizophila, Variovorax paradoxus. This result showed the biodiversity of endophytic diazotrophs from tuber and root crops. Of the 219 endophytic diazotrophs, 77 strains are identified as 23 species of Bacillus, and 29 strains are identified as 10 species of Pseudomonas. This makes up 106 strains of 33 species, in percentages of 48.40% and 41.77% of the 219 endophytic diazotrophs and 79 identified species. Indicating that Bacillus and Pseudomonas are dominant populations of endophytic diazotrophs from tuber and root crops. Conducted with 79 representatives of the 219 strains, PGP test showed that 8.86% strains showed ACC deaminase activity ranging from 0.026 to 13.76 μmol α-ketobutyrate·mg-1 protein·h-1, 64.56% strains showed IAA production ranging from 0.34 to 28.99 μg·mL-1, and 6.33%-13.92% strains showed antagonistic against phytopathogen Fusarium sporotrichioides ACCC37402, Fusarium xysporum MLS1 and Fusarium xysporum ACCC37438 with antifungal indexes of 41% to 63%. [Conclusion] Large number of endophytic diazotrophs habitat in the tuber and root of normally growing tuber and root crops. Endophytic diazotrophs from tuber and root crops phylogenetically belong 79 species of 24 genera showing wide distribution and huge biodiversity. Bacillus and Pseudomonas are dominant populations of endophytic diazotrophs from tuber and root crops