Sample records for bordetella

  1. Characterization of fimbrial subunits from Bordetella species

    Mooi, F.R.; Heide, H.G.J. van der; Avest, A.R. ter; Welinder, K.G.; Livey, I.; Zeijst, B.A.M. van der; Gaastra, W.


    Using antisera raised against serotype 2 and 3 fimbrial subunits from Bordetella pertussis, serologically related polypeptides were detected in Bordetella bronchiseptica, Bordetella parapertussis and Bordetella avium strains. The two B. pertussis fimbrial subunits, and three of the serologically rel

  2. Laboratory Maintenance of Bordetella pertussis.

    Hulbert, Robin R; Cotter, Peggy A


    The causative agent of the respiratory disease whooping cough, Bordetella pertussis, is a nutritionally fastidious microorganism but can be grown with relative ease in research laboratories. Stainer-Scholte synthetic broth medium and Bordet-Gengou blood agar both support growth of B. pertussis and are commonly used. B. pertussis prefers aerobic conditions and a temperature range of 35 degrees to 37 degrees C. Appropriate laboratory safety protocols are required to prevent the generation of aerosols, which could potentially spread this highly infectious agent. PMID:19885941

  3. Development of a PCR assay for identification of Bordetella hinzii

    Bordetella hinzii infects primarily poultry and immunocompromised humans. Although initially thought to be nonpathogenic in poultry, it was recently shown that some strains cause disease in turkey poults indistinguishable from the clinical presentation of turkey coryza caused by Bordetella avium. ...

  4. Bordetella pertussis diagnosed by polymerase chain reaction

    Birkebaek, N H; Heron, I; Skjødt, K


    The object of this work was to test the polymerase chain reaction (PCR) for demonstration of Bordetella pertussis (BP) in nasopharyngeal secretions. The method was applied to patients with recently diagnosed pertussis, as verified by BP culture. In order to test the sensitivity and specificity of...

  5. The Bordetella bhu Locus Is Required for Heme Iron Utilization

    Vanderpool, Carin K.; Armstrong, Sandra K.


    Bordetella pertussis and Bordetella bronchiseptica are capable of obtaining iron from hemin and hemoglobin. Genes encoding a putative bacterial heme iron acquisition system (bhu, for Bordetella heme utilization) were identified in a B. pertussis genomic sequence database, and the corresponding DNA was isolated from a virulent strain of B. pertussis. A B. pertussis bhuR mutant, predicted to lack the heme outer membrane receptor, was generated by allelic exchange. In contrast to the wild-type s...

  6. Phenotypic variation and modulation in Bordetella bronchiseptica.

    Peppler, M S; Schrumpf, M E


    Most of the isolates of Bordetella bronchiseptica obtained by this laboratory possessed a characteristic colonial morphology when grown on Bordet- Gengou agar (BGA) at 37 degrees C. The colonies appeared domed (Dom+) with a smooth colonial surface (Scs+) and a clear zone of hemolysis ( Hly +). From these Dom+ Scs+ Hly + BGA colony types arose flat (Dom-), smooth colonial surface (Scs+) and nonhemolytic ( Hly -) variants at frequencies of 10(-2) to 10(-3). Isogenic pairs of Dom+ Scs+ Hly + and...

  7. A PCR assay for identification of Bordetella hinzii

    Bordetella hinzii infects primarily poultry and immunocompromised humans. Although initially thought to be nonpathogenic in poultry, it was recently shown that some strains cause disease in turkey poults indistinguishable from the clinical presentation of turkey coryza caused by Bordetella avium. B....

  8. Bordetella bronchiseptica and fatal pneumonia of dogs and cats

    Bordetella bronchiseptica frequently causes nonfatal tracheobronchitis, but its role in fatal pneumonia is less well-studied. The objectives of this study were to identify the frequency of Bordetella bronchiseptica infection in fatal cases of bronchopneumonia in dogs and cats and to compare the diag...

  9. BpsR Modulates Bordetella Biofilm Formation by Negatively Regulating the Expression of the Bps Polysaccharide

    Conover, Matt S.; Redfern, Crystal J.; Ganguly, Tridib; Sukumar, Neelima; Sloan, Gina; Mishra, Meenu; Deora, Rajendar


    Bordetella bacteria are Gram-negative respiratory pathogens of animals, birds, and humans. A hallmark feature of some Bordetella species is their ability to efficiently survive in the respiratory tract even after vaccination. Bordetella bronchiseptica and Bordetella pertussis form biofilms on abiotic surfaces and in the mouse respiratory tract. The Bps exopolysaccharide is one of the critical determinants for biofilm formation and the survival of Bordetella in the murine respiratory tract. In...

  10. Genomic island excisions in Bordetella petrii

    Levillain Erwan


    Full Text Available Abstract Background Among the members of the genus Bordetella B. petrii is unique, since it is the only species isolated from the environment, while the pathogenic Bordetellae are obligately associated with host organisms. Another feature distinguishing B. petrii from the other sequenced Bordetellae is the presence of a large number of mobile genetic elements including several large genomic regions with typical characteristics of genomic islands collectively known as integrative and conjugative elements (ICEs. These elements mainly encode accessory metabolic factors enabling this bacterium to grow on a large repertoire of aromatic compounds. Results During in vitro culture of Bordetella petrii colony variants appear frequently. We show that this variability can be attributed to the presence of a large number of metastable mobile genetic elements on its chromosome. In fact, the genome sequence of B. petrii revealed the presence of at least seven large genomic islands mostly encoding accessory metabolic functions involved in the degradation of aromatic compounds and detoxification of heavy metals. Four of these islands (termed GI1 to GI3 and GI6 are highly related to ICEclc of Pseudomonas knackmussii sp. strain B13. Here we present first data about the molecular characterization of these islands. We defined the exact borders of each island and we show that during standard culture of the bacteria these islands get excised from the chromosome. For all but one of these islands (GI5 we could detect circular intermediates. For the clc-like elements GI1 to GI3 of B. petrii we provide evidence that tandem insertion of these islands which all encode highly related integrases and attachment sites may also lead to incorporation of genomic DNA which originally was not part of the island and to the formation of huge composite islands. By integration of a tetracycline resistance cassette into GI3 we found this island to be rather unstable and to be lost from

  11. Epithelial cell invasion and survival of Bordetella bronchiseptica.

    SCHIPPER, H; Krohne, G F; R. Gross


    Wild-type Bordetella bronchiseptica and a bvg mutant strain were used for invasion and survival experiments in human Caco-2 and A549 epithelial cells. Both bacterial strains were able to enter and persist within the host cells for at least a week. A significant proportion of the bacteria from both B. bronchiseptica strains but not from Bordetella pertussis were found free in the cytoplasm, suggesting different invasion and survival strategies of the two species in epithelial cells.

  12. Extended incubation of culture plates improves recovery of Bordetella spp.

    Katzko, G; Hofmeister, M; Church, D.


    Extended incubation of culture plates was studied to see if the recovery of Bordetella spp. from nasopharyngeal swabs could be improved. Forty-eight Bordetella isolates were recovered from 103 children (overall positive-culture rate, 46.6%) who met the clinical case definition of pertussis. Seven of 44 (16%) B. pertussis isolates and 2 of 4 (50%) B. parapertussis isolates were recovered only after extended incubation of nasopharyngeal cultures up to 12 days.

  13. Prevalence of Bordetella pertussis and Bordetella parapertussis in Samples Submitted for RSV Screening

    Walsh, Paul


    Full Text Available BACKGROUND: The clinical presentation of Bordetella pertussis can overlap with that of respiratory syncytial virus (RSV; however, management differs.HYPOTHESIS: First, the prevalence of B. pertussis is less than 2% among patients screened for RSV, and second the prevalence of B. parapertussis is also less than 2% among these patients.METHODS: Nasal washings submitted to a clinical laboratory for RSV screening were tested for B. pertussis and B. parapertussis, using species-specific real-time polymerase chain reaction (PCR assays. These were optimized to target conserved regions within a complement gene and the CarB gene, respectively. A Bordetella spp. genus-specific real-time PCR assay was designed to detect the Bhur gene of B. pertussis, B. parapertussis, and B. bronchiseptica. RSV A and B subtypes were tested by reverse transcription-PCR.RESULTS: Four hundred and eighty-nine clinical samples were tested. There was insufficient material to complete testing for one B. pertussis, 10 RSV subtype A, and four RSV subtype B assays. Bordetella pertussis was detected in 3/488 (0.6% (95% CI 0.1% to 1.8%, while B. parapertussis was detected in 5/489 (1.0% (95% CI 0.3% to 2.4%. Dual infection of B. pertussis with RSV and of B. parapertussis with RSV occurred in two and in three cases respectively. RSV was detected by PCR in 127 (26.5%.CONCLUSION: The prevalence of B. pertussis in nasal washings submitted for RSV screening was less than 2%. The prevalence of parapertussis may be higher than 2%. RSV with B. pertussis and RSV with B. parapertussis coinfection do occur.

  14. Occurrence of Bordetella infection in pigs in northern India.

    Kumar, Sandeep; Singh, Bhoj R; Bhardwaj, Monika; Singh, Vidya


    Bordetella bronchiseptica infection causing atrophic rhinitis in pigs is reported from almost all countries. In the present study, occurrence of Bordetella infection in apparently healthy pigs was determined in 392 pigs sampled to collect 358 serum samples and 316 nasal swabs from Northern India by conventional bacterioscopy, detection of antigen with multiplex polymerase chain reaction (mPCR), and detection of antibodies with microagglutination test (MAT) and enzyme linked immune-sorbent assay (ELISA). Bordetella bronchiseptica could be isolated from six (1.92%) nasal swabs. Although isolates varied significantly in their antimicrobial sensitivity, they had similar plasmid profile. The genus specific and species specific amplicons were detected from 8.2% and 4.4% nasal swabs using mPCR with alc gene (genus specific) and fla gene and fim2 gene (species specific) primers, respectively. Observations revealed that there may be other bordetellae infecting pigs because about 50% of the samples positive using mPCR for genus specific amplicons failed to confirm presence of B. bronchiseptica. Of the pig sera tested with MAT and ELISA for Bordetella antibodies, 67.6% and 86.3% samples, respectively, were positive. For antigen detection mPCR was more sensitive than conventional bacterioscopy while for detection of antibodies neither of the two tests (MAT and ELISA) had specificity in relation to antigen detection. Study indicated high prevalence of infection in swine herds in Northern India. PMID:24688547

  15. Occurrence of Bordetella Infection in Pigs in Northern India

    Sandeep Kumar


    Full Text Available Bordetella bronchiseptica infection causing atrophic rhinitis in pigs is reported from almost all countries. In the present study, occurrence of Bordetella infection in apparently healthy pigs was determined in 392 pigs sampled to collect 358 serum samples and 316 nasal swabs from Northern India by conventional bacterioscopy, detection of antigen with multiplex polymerase chain reaction (mPCR, and detection of antibodies with microagglutination test (MAT and enzyme linked immune-sorbent assay (ELISA. Bordetella bronchiseptica could be isolated from six (1.92% nasal swabs. Although isolates varied significantly in their antimicrobial sensitivity, they had similar plasmid profile. The genus specific and species specific amplicons were detected from 8.2% and 4.4% nasal swabs using mPCR with alc gene (genus specific and fla gene and fim2 gene (species specific primers, respectively. Observations revealed that there may be other bordetellae infecting pigs because about 50% of the samples positive using mPCR for genus specific amplicons failed to confirm presence of B. bronchiseptica. Of the pig sera tested with MAT and ELISA for Bordetella antibodies, 67.6% and 86.3% samples, respectively, were positive. For antigen detection mPCR was more sensitive than conventional bacterioscopy while for detection of antibodies neither of the two tests (MAT and ELISA had specificity in relation to antigen detection. Study indicated high prevalence of infection in swine herds in Northern India.

  16. Strain-specific virulence of Bordetella hinzii in poultry

    Two species of Bordetella, B. avium and B. hinzii, are known to infect avian hosts. B. avium is the etiologic agent of turkey coryza, a disease of high morbidity. B. hinzii, though commonly acquired from the respiratory tracts of diseased poultry, has not been demonstrated to be pathogenic in eith...

  17. Bordetella bronchiseptica phase variation induced by crystal violet.

    Ishikawa, H.; Isayama, Y


    A method for effective induction of phase variation in Bordetella bronchiseptica by treatment with crystal violet (CV) is presented. When grown in CV-broth, phase I cells dissociated into three serial phases. Appearance of variant cells was observed simultaneously with the beginning of cell multiplication. The maximum effect of CV was obtained at a concentration of 8 micrograms/ml, when the proportion of variants in the population reached 100%. The main factors which affected phase variation ...

  18. Bordetella avium Antibiotic Resistance, Novel Enrichment Culture, and Antigenic Characterization

    Beach, Nathan M; Thompson, Seth; Mutnick, Rachel; Brown, Lisa; Kettig, Gina; Puffenbarger, Robyn; Miyamoto, David; Temple, Louise


    Bordetella avium continues to be an economic issue in the turkey industry as the causative agent of bordetellosis, which often leads to serious secondary infections. This study presents a broad characterization of the antibiotic resistance patterns in this diverse collection of B. avium strains collected over the past thirty years. In addition, the plasmid basis for the antibiotic resistance was characterized. The antibiotic resistance pattern allowed the development of a novel enrichment cul...

  19. Properties of dermonecrotic toxin prepared from sonic extracts Bordetella bronchiseptica.

    Kume, K.; Nakai, T.; Samejima, Y; Sugimoto, C


    A toxin with dermonecrotic activity (DNT) was purified from sonic extracts of Bordetella bronchiseptica L3 of pig origin at phase I by chromatographic and electrophoretic methods. The purification procedure was one developed for obtaining the Pasteurella multocida DNT from sonic extracts with some modifications. Dermonecrotizing activity of B. bronchiseptica-purified DNT was increased by 600-fold compared with that of the crude extract, and the average yield was about 3%. The toxin was homoge...

  20. Cilia-associated bacteria in fatal Bordetella bronchiseptica pneumonia of dogs and cats

    Bordetella bronchiseptica frequently causes nonfatal tracheobronchitis, but its role in fatal pneumonia is less well-studied. The objectives of this study were to identify the frequency of Bordetella bronchiseptica infection in fatal cases of bronchopneumonia in dogs and cats and to compare the diag...

  1. [Bordetella bronchiseptica recurrent bacteraemia in a patient with bone marrow transplantation].

    Echeverri-Toro, Lina; Arango, Andrés; Ospina, Sigifredo; Agudelo, Carlos


    We report a case of recurrent bacteraemia caused by Bordetella bronchiseptica in an immunocompromised patient with a history of allogenic bone marrow transplantation for myelodysplastic syndrome, who was admitted to hospital with febrile syndrome. Bordetella bronchiseptica is an uncommon human pathogen which mainly affects immunocompromised patients, being a rare cause of bacteraemia. PMID:26849691

  2. The missing link: Bordetella petrii is endowed with both the metabolic versatility of environmental bacteria and virulence traits of pathogenic Bordetellae

    Schneiker-Bekel Susanne


    Full Text Available Abstract Background Bordetella petrii is the only environmental species hitherto found among the otherwise host-restricted and pathogenic members of the genus Bordetella. Phylogenetically, it connects the pathogenic Bordetellae and environmental bacteria of the genera Achromobacter and Alcaligenes, which are opportunistic pathogens. B. petrii strains have been isolated from very different environmental niches, including river sediment, polluted soil, marine sponges and a grass root. Recently, clinical isolates associated with bone degenerative disease or cystic fibrosis have also been described. Results In this manuscript we present the results of the analysis of the completely annotated genome sequence of the B. petrii strain DSMZ12804. B. petrii has a mosaic genome of 5,287,950 bp harboring numerous mobile genetic elements, including seven large genomic islands. Four of them are highly related to the clc element of Pseudomonas knackmussii B13, which encodes genes involved in the degradation of aromatics. Though being an environmental isolate, the sequenced B. petrii strain also encodes proteins related to virulence factors of the pathogenic Bordetellae, including the filamentous hemagglutinin, which is a major colonization factor of B. pertussis, and the master virulence regulator BvgAS. However, it lacks all known toxins of the pathogenic Bordetellae. Conclusion The genomic analysis suggests that B. petrii represents an evolutionary link between free-living environmental bacteria and the host-restricted obligate pathogenic Bordetellae. Its remarkable metabolic versatility may enable B. petrii to thrive in very different ecological niches.

  3. Suppression of Platelet Aggregation by Bordetella pertussis Adenylate Cyclase Toxin

    Iwaki, Masaaki; Kamachi, Kazunari; Heveker, Nikolaus; Konda, Toshifumi


    The effect of Bordetella pertussis adenylate cyclase toxin (ACT) on platelet aggregation was investigated. This cell-invasive adenylate cyclase completely suppressed ADP (10 μM)-induced aggregation of rabbit platelets at 3 μg/ml and strongly suppressed thrombin (0.2 U/ml)-induced aggregation at 10 μg/ml. The suppression was accompanied by marked increase in platelet intracellular cyclic AMP (cAMP) content and was diminished by the anti-ACT monoclonal antibody B7E11. A catalytically inactive p...

  4. Polymorphisms influencing expression of dermonecrotic toxin in Bordetella bronchiseptica.

    Keisuke Okada

    Full Text Available Bordetella bronchiseptica is a pathogenic bacterium causing respiratory infections in a broad range of mammals. Recently, we determined the whole genome sequence of B. bronchiseptica S798 strain isolated from a pig infected with atrophic rhinitis and found four single-nucleotide polymorphisms (SNPs at positions -129, -72, +22, and +38 in the region upstream of dnt encoding dermonecrotic toxin (DNT, when compared with a rabbit isolate, RB50. DNT is known to be involved in turbinate atrophy observed in atrophic rhinitis. Immunoblotting, quantitative real-time PCR, and β-galactosidase reporter assay revealed that these SNPs resulted in the increased promoter activity of dnt and conferred the increased ability to produce DNT on the bacteria. Similar or identical SNPs were also found in other pig isolates kept in our laboratory, all of which produce a larger amount of DNT than RB50. Our analysis revealed that substitution of at least two of the four bases, at positions -72 and +22, influenced the promoter activity for dnt. These results imply that these SNPs are involved in the pathogenicity of bordetellae specific to pig diseases.

  5. Identification and characterization of iron-regulated Bordetella pertussis alcaligin siderophore biosynthesis genes.

    Kang, H.Y.; Brickman, T J; Beaumont, F C; Armstrong, S K


    Bordetella bronchiseptica mutants BRM1, BRM6, and BRM9 fail to produce the native dihydroxamate siderophore alcaligin. A 4.5-kb BamHI-Smal Bordetella pertussis genomic DNA fragment carried multiple genes required to restore alcaligin production to these siderophore-deficient mutants. Phenotypic complementation analysis using subclones of the 4.5-kb genomic region demonstrated that the closely linked BRM1 and BRM9 mutations were genetically separable from the BRM6 mutation, and both insertions...

  6. Real-time PCR-based detection of Bordetella pertussis and Bordetella parapertussis in an Irish paediatric population.

    Grogan, Juanita A


    Novel real-time PCR assays targeting the Bordetella pertussis insertion sequence IS481, the toxin promoter region and Bordetella parapertussis insertion sequence IS1001 were designed. PCR assays were capable of detecting ≤10 copies of target DNA per reaction, with an amplification efficiency of ≥90 %. From September 2003 to December 2009, per-nasal swabs and nasopharyngeal aspirates submitted for B. pertussis culture from patients ≤1 month to >15 years of age were examined by real-time PCR. Among 1324 patients, 76 (5.7 %) were B. pertussis culture positive and 145 (10.95 %) were B. pertussis PCR positive. Of the B. pertussis PCR-positive patients, 117 (81 %) were aged 6 months or less. A total of 1548 samples were examined, of which 87 (5.6 %) were culture positive for B. pertussis and 169 (10.92 %) were B. pertussis PCR positive. All culture-positive samples were PCR positive. Seven specimens (0.5 %) were B. parapertussis culture positive and 10 (0.8 %) were B. parapertussis PCR positive, with all culture-positive samples yielding PCR-positive results. A review of patient laboratory records showed that of the 1324 patients tested for pertussis 555 (42 %) had samples referred for respiratory syncytial virus (RSV) testing and 165 (30 %) were positive, as compared to 19.4 % of the total 5719 patients tested for RSV in this period. Analysis of the age distribution of RSV-positive patients identified that 129 (78 %) were aged 6 months or less, similar to the incidence observed for pertussis in that patient age group. In conclusion, the introduction of the real-time PCR assays for the routine detection of B. pertussis resulted in a 91 % increase in the detection of the organism as compared to microbiological culture. The incidence of infection with B. parapertussis is low while the incidence of RSV infection in infants suspected of having pertussis is high, with a similar age distribution to B. pertussis infection.

  7. Bordetella avium antibiotic resistance, novel enrichment culture, and antigenic characterization.

    Beach, Nathan M; Thompson, Seth; Mutnick, Rachel; Brown, Lisa; Kettig, Gina; Puffenbarger, Robyn; Stockwell, Stephanie B; Miyamoto, David; Temple, Louise


    Bordetella avium continues to be an economic issue in the turkey industry as the causative agent of bordetellosis, which often leads to serious secondary infections. This study presents a broad characterization of the antibiotic resistance patterns in this diverse collection of B. avium strains collected over the past thirty years. In addition, the plasmid basis for the antibiotic resistance was characterized. The antibiotic resistance pattern allowed the development of a novel enrichment culture method that was subsequently employed to gather new isolates from diseased turkeys and a healthy sawhet owl. While a healthy turkey flock was shown to seroconvert by four weeks-of-age, attempts to culture B. avium from healthy turkey poults were unsuccessful. Western blot of B. avium strains using pooled serum from diseased and healthy commercial turkey flocks revealed both antigenic similarities and differences between strains. In sum, the work documents the continued exposure of commercial turkey flocks to B. avium and the need for development of an effective, inexpensive vaccine to control spread of the disease. PMID:22721730

  8. New Data on Vaccine Antigen Deficient Bordetella pertussis Isolates

    Valérie Bouchez


    Full Text Available Evolution of Bordetella pertussis is driven by natural and vaccine pressures. Isolates circulating in regions with high vaccination coverage present multiple allelic and antigenic variations as compared to isolates collected before introduction of vaccination. Furthermore, during the last epidemics reported in regions using pertussis acellular vaccines, isolates deficient for vaccine antigens, such as pertactin (PRN, were reported to reach high proportions of circulating isolates. More sporadic filamentous hemagglutinin (FHA or pertussis toxin (PT deficient isolates were also collected. The whole genome of some recent French isolates, deficient or non-deficient in vaccine antigens, were analyzed. Transcription profiles of the expression of the main virulence factors were also compared. The invasive phenotype in an in vitro human tracheal epithelial (HTE cell model of infection was evaluated. Our genomic analysis focused on SNPs related to virulence genes known to be more likely to present allelic polymorphism. Transcriptomic data indicated that isolates circulating since the introduction of pertussis vaccines present lower transcription levels of the main virulence genes than the isolates of the pre-vaccine era. Furthermore, isolates not producing FHA present significantly higher expression levels of the entire set of genes tested. Finally, we observed that recent isolates are more invasive in HTE cells when compared to the reference strain, but no multiplication occurs within cells.

  9. Early Induction of Cytokines in Pigs Coinfected with Swine Influenza Virus and Bordetella bronchiseptica

    Respiratory disease is one of the most important health issues for the swine industry, and coinfection with two or more pathogens is a common occurrence. Bordetella bronchiseptica and swine influenza virus (SIV) are important and common respiratory pathogens of pigs. The effect of coinfection of S...

  10. Filamentous hemagglutinin of Bordetella pertussis: a key adhesin with immunomodulatory properties?

    Villarino Romero, Rodrigo; Osička, Radim; Šebo, Peter


    Roč. 9, č. 12 (2014), s. 1339-1360. ISSN 1746-0913 R&D Projects: GA ČR(CZ) P302/11/0580; GA ČR(CZ) GA13-14547S Institutional support: RVO:61388971 Keywords : Bordetella * adhesion * integrins * filamentous hemagglutinin Subject RIV: EE - Microbiology, Virology Impact factor: 4.275, year: 2014

  11. Severe infantile Bordetella pertussis pneumonia in monozygotic twins with a congenital C3 deficiency

    Kurvers, R.A.J.; Westra, D.; Heijst, A.F.J. van; Walk, T.L.M.; Warris, A.; Kar, N.C.A.J. van de


    Bordetella pertussis or whooping cough is a vaccine-preventable disease that still remains a serious infection in neonates and young infants. We describe two young infants, monozygotic twins, with a severe B. pertussis pneumonia of whom one needed extracorporeal membrane oxygenation. Diagnostic work

  12. Identification of Bordetella bronchseptica in fatal pneumonia of dogs and cats

    Infection with Bordetella bronchiseptica is a common cause of tracheobronchitis and upper respiratory disease in dogs and cats, but it can also lead to fatal pneumonia. Identification of this pathogen is important due the risk of transmission to other animals, availability of vaccines and potential...

  13. SNP-based typing: a useful tool to study Bordetella pertussis populations

    Gent, M. van; Bart, M.J.; Heide, H.G. van der; Heuvelman, K.J.; Kallonen, T.; He, Q.; Mertsola, J.; Advani, A.; Hallander, H.O.; Janssens, K.; Hermans, P.W.M.; Mooi, F.R.


    To monitor changes in Bordetella pertussis populations, mainly two typing methods are used; Pulsed-Field Gel Electrophoresis (PFGE) and Multiple-Locus Variable-Number Tandem Repeat Analysis (MLVA). In this study, a single nucleotide polymorphism (SNP) typing method, based on 87 SNPs, was developed a

  14. Attenuated Bordetella pertussis Vaccine Protects against Respiratory Syncytial Virus Disease via an IL-17-Dependent Mechanism

    Sawant, Devika; Schnoeller, Corinna; Roux, Xavier; Openshaw, Peter J.; Olszewska, Wieslawa; Locht, Camille; Raze, Dominique


    Rationale: We attenuated virulent Bordetella pertussis by genetically eliminating or detoxifying three major toxins. This strain, named BPZE1, is being developed as a possible live nasal vaccine for the prevention of whooping cough. It is immunogenic and safe when given intranasally in adult volunteers.

  15. The Bordetella pertussis protein Pertactin: role in immunity and immune evasion

    Hijnen, Marcel


    Pertussis is a highly contagious infectious disease of the respiratory tract which is caused by Bordetella pertussis. Before widespread introduction of vaccination against pertussis, almost every child contracted pertussis. The disease is most severe in neonates and children under the age of 1. Intr

  16. Bordetella bronchiseptica associated with pulmonary disease in mountain voles (Microtus montanus)

    Jensen, W.I.; Duncan, R.M.


    Bordetella bronchiseptica was isolated from the lungs of all of six mountain voles (Microtus montanus) found dead or dying of pulmonary infection near the Bear River Research Station in northern Utah in January, 1973. The possibility of concomitant viral or mycoplasmal infection was not ruled out.

  17. Evidence for horizontal gene transfer of two antigenically distinct O antigens in Bordetella bronchiseptica

    Antigenic variation is one mechanism pathogens use to avoid immune-mediated competition between closely related strains. Here, we show that two Bordetella bronchiseptica strains, RB50 and 1289, express two antigenically distinct O-antigen serotypes (O1 and O2 respectively). When 18 additional B. b...

  18. Canine distemper virus infection with secondary Bordetella bronchiseptica pneumonia in dogs Infecção pelo virus da cinomose com pneumonia secundária por Bordetella bronchiseptica em cães

    Selwyn Arlington Headley; Dominguita Lühers Graça; Mateus Matiuzzi da Costa; Agueda Castagna de Vargas


    Canine distemper virus infection and secondary Bordetella bronchiseptica pneumonia are described in mongrel dogs. Canine distemper was characterised by nonsuppurative demyelinating encephalitis with typical inclusion bodies in astrocytes. B. bronchiseptica was isolated from areas of purulent bronchopneumonia.São descritas as infecções simultâneas do vírus da cinomose canina e Bordetella bronchiseptica em caninos sem raça definida. As lesões de cinomose foram caracterizadas por encefalite desm...

  19. Bordetella pertussis acquires resistance to complement-mediated killing in vivo.

    Pishko, Elizabeth J; Betting, David J; Hutter, Christina S; Harvill, Eric T


    In order to initially colonize a host, bacteria must avoid various components of the innate immune system, one of which is complement. The genus Bordetella includes three closely related species that differ in their ability to resist complement-mediated killing. Bordetella parapertussis and Bordetella bronchiseptica resist killing in naïve serum, a characteristic that may aid in efficient respiratory tract colonization and has been attributed to expression of O antigen. Bordetella pertussis lacks O antigen and is sensitive to naïve serum in vitro, yet it also efficiently colonizes the respiratory tract. Based on these observations, we hypothesized that B. pertussis may have an alternate mechanism to resist complement in vivo. While a number of reports on serum sensitivity of the bordetellae have been published, we show here that serum concentration and growth conditions can greatly alter the observed level of sensitivity to complement and that all but one strain of B. pertussis observed were sensitive to some level of naïve serum in vitro, particularly when there was excess complement. However, B. pertussis rapidly acquires increased resistance in vivo to naïve serum that is specific to the alternative pathway. Resistance is not efficiently acquired by B. parapertussis and B. bronchiseptica mutants lacking O antigen. This B. pertussis-specific mechanism of complement resistance does not appear to be dependent on either brkA or other genes expressed specifically in the Bvg(+) phase. This in vivo acquisition of alternative pathway resistance suggests that there is a novel O antigen-independent method by which B. pertussis evades complement-mediated killing. PMID:12933835

  20. Extracellular DNA Is Essential for Maintaining Bordetella Biofilm Integrity on Abiotic Surfaces and in the Upper Respiratory Tract of Mice

    Conover, Matt S.; Mishra, Meenu; Deora, Rajendar


    Bacteria form complex and highly elaborate surface adherent communities known as biofilms which are held together by a self-produced extracellular matrix. We have previously shown that by adopting a biofilm mode of existence in vivo, the Gram negative bacterial pathogens Bordetella bronchiseptica and Bordetella pertussis are able to efficiently colonize and persist in the mammalian respiratory tract. In general, the bacterial biofilm matrix includes polysaccharides, proteins and extracellular...

  1. Bordetella pertussis en estudiantes adolescentes de la Ciudad de México Bordetella pertussis em estudantes adolescentes da Cidade do México Bordetella pertussis in adolescents students in Mexico City

    Patricia Tomé Sandoval


    Full Text Available OBJETIVO: Estimar la seroprevalencia a Bordetella pertussis en escolares y sus contactos escolares y familiares. MÉTODOS: Un total de 12.273 estudiantes de 12 a 15 años de edad, de 14 escuelas secundarias públicas de la Ciudad de México fueron estudiados durante los meses de Septiembre 2002 a Marzo 2003. Se tomó muestra de exudado nasofaríngeo en adolescentes con tos de más de 14 días de evolución. La infección fue confirmada por la técnica de reacción en cadena de polimerasa. Se realizó estudio de contactos escolares y familiares. RESULTADOS: La incidencia de tos fue de 5 para 1.000 estudiantes. De los 61 estudiantes con tos incluidos en la muestra, 20 (32,8% fueron positivos para Bordetella. De los 152 contactos escolares, 16 (10,6% resultaron positivos, y ocho tenían tos. Uno de esos contactos fue el director de una de las escuelas responsable de más del 60% de los casos positivos (12/20, quien también dio lecciones a diez de los estudiantes infectados. De los 29 familiares, ocho (27,6% fueron positivos, pertenecientes a tres familias. CONCLUSIONES: Los resultados muestran que la frecuencia de la enfermedad fue similar al comunicado en la población adolescente de otros países. Sin embargo, este trastorno no tiene necesariamente signos clínicos de la tos persistente y está sujeto a la existencia de infectados asintomáticos con Bordetella.OBJETIVO: Estimar a soroprevalência a Bordetella pertussis em escolares e seus contatos. MÉTODOS: Foram examinados 12.273 alunos entre 12 e 15 anos de idade, de 14 escolas secundárias públicas da Cidade do México, de setembro de 2002 a março de 2003. Amostras de exudado nasofaríngeo foram coletadas de adolescentes com tosse por mais de 14 dias. A infecção foi confirmada por reação em cadeia da polimerase. Todos os alunos e funcionários dos colégios dos casos confirmados por reação em cadeia da polimerase e seus familiares foram testados. RESULTADOS: A incidência de tosse

  2. Analysis of Bordetella pertussis clinical isolates circulating in European countries during the period 1998–2012

    Gent, M.; Heuvelman, C.J.; van der Heide, H G; Hallander, H. O.; Advani, A.; Guiso, N; Wirsing von Kőnig, C. H.; Vestrheim, D. F.; Dalby, T.; Fry, N. K.; Pierard, D; Detemmerman, L; Zavadilova, J.; Fabianova, K.; Logan, C.


    Despite more than 50 years of vaccination, pertussis is still an endemic disease, with regular epidemic outbreaks. With the exception of Poland, European countries have replaced whole-cell vaccines (WCVs) by acellular vaccines (ACVs) in the 1990s. Worldwide, antigenic divergence in vaccine antigens has been found between vaccine strains and circulating strains. In this work, 466 Bordetella pertussis isolates collected in the period 1998–2012 from 13 European countries were characterised by mu...

  3. Recovery of Bordetella pertussis from PCR-positive nasopharyngeal samples is dependent on bacterial load.

    Vestrheim, Didrik F; Steinbakk, Martin; Bjørnstad, Martha L; Moghaddam, Amir; Reinton, Nils; Dahl, Mette L; Grude, Nils; Sandven, Per


    Viable Bordetella pertussis isolates are essential for surveillance purposes. We performed culture of 223 PCR-positive nasopharyngeal samples. B. pertussis was recovered from 45 (20.2%) of the samples. Growth was associated with a high bacterial load, as determined by PCR. Culture from PCR-positive samples is a feasible approach to recover B. pertussis isolates, and culture can be limited to samples with a high bacterial load. PMID:23035189

  4. Recovery of Bordetella pertussis from PCR-Positive Nasopharyngeal Samples Is Dependent on Bacterial Load

    Vestrheim, Didrik F.; Steinbakk, Martin; Bjørnstad, Martha L.; Moghaddam, Amir; Reinton, Nils; Dahl, Mette L.; Grude, Nils; Sandven, Per


    Viable Bordetella pertussis isolates are essential for surveillance purposes. We performed culture of 223 PCR-positive nasopharyngeal samples. B. pertussis was recovered from 45 (20.2%) of the samples. Growth was associated with a high bacterial load, as determined by PCR. Culture from PCR-positive samples is a feasible approach to recover B. pertussis isolates, and culture can be limited to samples with a high bacterial load.

  5. Monoclonal Antibodies Directed Against the Outer Membrane Protein of Bordetella avium

    Liu, Guanhua; Liang, Manfei; Zuo, Xuemei; Zhao, Xue; Guo, Fanxia; Yang, Shifa; Zhu, Ruiliang


    Bordetella avium is the etiologic agent of coryza and rhinotracheitis in poultry. This respiratory disease is responsible for substantial economic losses in the poultry industry. Monoclonal antibodies (MAbs) were produced against the outer membrane proteins (OMPs) of B. avium isolated from diseased chickens. BALB/c mice were immunized with the extracted B. avium OMPs. Then the splenocytes from immunized mice and SP2/0 myeloma cells were fused using PEG 4000. Three stable hybridoma clones (des...

  6. Host Specificity of Ovine Bordetella parapertussis and the Role of Complement.

    Sara E Hester

    Full Text Available The classical bordetellae are comprised of three subspecies that differ from broad to very limited host specificity. Although several lineages appear to have specialized to particular host species, most retain the ability to colonize and grow in mice, providing a powerful common experimental model to study their differences. One of the subspecies, Bordetella parapertussis, is composed of two distinct clades that have specialized to different hosts: one to humans (Bpphu, and the other to sheep (Bppov. While Bpphu and the other classical bordetellae can efficiently colonize mice, Bppov strains are severely defective in their ability to colonize the murine respiratory tract. Bppov genomic analysis did not reveal the loss of adherence genes, but substantial mutations and deletions of multiple genes involved in the production of O-antigen, which is required to prevent complement deposition on B. bronchiseptica and Bpphu strains. Bppov lacks O-antigen and, like O-antigen mutants of other bordetellae, is highly sensitive to murine complement-mediated killing in vitro. Based on these results, we hypothesized that Bppov failed to colonize mice because of its sensitivity to murine complement. Consistent with this, the Bppov defect in the colonization of wild type mice was not observed in mice lacking the central complement component C3. Furthermore, Bppov strains were highly susceptible to killing by murine complement, but not by sheep complement. These data demonstrate that the failure of Bppov to colonize mice is due to sensitivity to murine, but not sheep, complement, providing a mechanistic example of how specialization that accompanies expansion in one host can limit host range.

  7. Detection of Bordetella pertussis by rapid-cycle PCR and colorimetric microwell hybridization.

    Buck, G E


    The use of rapid-cycle PCR combined with colorimetric microwell hybridization for detecting Bordetella pertussis was investigated. Rapid cycling was performed with an air thermocycler (model 1605; Idaho Technology, Idaho Falls, Idaho). Although the instrument was originally designed to be used with capillary tubes, an adapter that allows this instrument to be used with PCR tubes has recently been introduced. Because of the low heat capacity of air, the thermocycler has rapid transition rates ...

  8. Detection of IgG antibodies against Bordetella pertussis with 125I-protein A

    A method for the detection of IgG antibodies against Bordetella pertussis is described, based on the principle of 'sandwich' radioimmunoassay. 125I protein A is used as radioactive tracer. The influence of amounts of antigen, antibody, radioactive tracer, incubation time and temperature were tested and the optimal conditions for the assay are described. The procedure offers a simple, quick, and sensitive method for detecting antibodies against B. pertussis. Application and limitation of the test are discussed. (orig.)

  9. Bordetella pertussis adenylate cyclase toxin translocation across a tethered lipid bilayer

    Veneziano, Rémi; Rossi, Claire; Chenal, Alexandre; Devoisselle, Jean-Marie; Ladant, Daniel; Chopineau, Joel


    Many bacterial toxins can cross biological membranes to reach the cytosol of mammalian cells, although how they pass through a lipid bilayer remains largely unknown. Bordetella pertussis adenylate cyclase (CyaA) toxin delivers its catalytic domain directly across the cell membrane. To characterize this unique translocation process, we designed an in vitro assay based on a tethered lipid bilayer assembled over a biosensor surface derivatized with calmodulin, a natural activator of the toxin. C...

  10. Mutations in Cytochrome Assembly and Periplasmic Redox Pathways in Bordetella pertussis

    Feissner, Robert E.; Beckett, Caroline S.; Loughman, Jennifer A.; Kranz, Robert G.


    Transposon mutagenesis of Bordetella pertussis was used to discover mutations in the cytochrome c biogenesis pathway called system II. Using a tetramethyl-p-phenylenediamine cytochrome c oxidase screen, 27 oxidase-negative mutants were isolated and characterized. Nine mutants were still able to synthesize c-type cytochromes and possessed insertions in the genes for cytochrome c oxidase subunits (ctaC, -D, and -E), heme a biosynthesis (ctaB), assembly of cytochrome c oxidase (sco2), or ferroch...

  11. Bordetella Bronchiseptica in the Immunosuppressed Population – A Case Series and Review

    Yacoub, Abraham T.; Katayama, Mitsuya; Tran, JoAnn; Zadikany, Ronit; Kandula, Manasa; Greene, John


    Organisms that are not known to cause serious infection in the immunocompetent population can, in fact, cause devastating illness in immunosuppressed neutropenic populations especially those who are undergoing hematopoietic stem cell transplantation (HSCT), and solid organ transplantation or a history of malignancy. One organism of interest isolated from immunosuppressed patients at our institution was Bordetella bronchiseptica. It is known to cause respiratory tract disease in the animal pop...

  12. Extracellular DNA is essential for maintaining Bordetella biofilm integrity on abiotic surfaces and in the upper respiratory tract of mice.

    Matt S Conover

    Full Text Available Bacteria form complex and highly elaborate surface adherent communities known as biofilms which are held together by a self-produced extracellular matrix. We have previously shown that by adopting a biofilm mode of existence in vivo, the gram negative bacterial pathogens Bordetella bronchiseptica and Bordetella pertussis are able to efficiently colonize and persist in the mammalian respiratory tract. In general, the bacterial biofilm matrix includes polysaccharides, proteins and extracellular DNA (eDNA. In this report, we investigated the function of DNA in Bordetella biofilm development. We show that DNA is a significant component of Bordetella biofilm matrix. Addition of DNase I at the initiation of biofilm growth inhibited biofilm formation. Treatment of pre-established mature biofilms formed under both static and flow conditions with DNase I led to a disruption of the biofilm biomass. We next investigated whether eDNA played a role in biofilms formed in the mouse respiratory tract. DNase I treatment of nasal biofilms caused considerable dissolution of the biofilm biomass. In conclusion, these results suggest that eDNA is a crucial structural matrix component of both in vitro and in vivo formed Bordetella biofilms. This is the first evidence for the ability of DNase I to disrupt bacterial biofilms formed on host organs.

  13. Molecular analysis of the bvg-repressed urease of Bordetella bronchiseptica.

    McMillan, D J; Shojaei, M; Chhatwal, G S; Guzmán, C A; Walker, M J


    Bordetella bronchiseptica is a ureolytic mammalian respiratory pathogen. We have investigated the regulation of urease in B. bronchiseptica and the potential role of this enzyme in eukaryotic invasion and intracellular survival. Our results indicate urease is a bordetella virulence repressed gene. Urease activity in virulent B. bronchiseptica BB7865 is up-regulated from basal levels by 5 gl1 magnesium sulphate at 37 degrees C. At 30 degrees C, urease activity remained at basal levels, even in the presence on magnesium sulphate, suggesting a second temperature dependent mechanism of urease regulation was also operating. Urease was not inducible by 10 mM urea nor up-regulated in nitrogen limiting conditions. To evaluate the role of urease in intracellular invasion and survival urease-negative mutants of B. bronchiseptica BB7865 and B. bronchiseptica BB7866 were created by transposon mutagenesis, and compared to the urease-positive parental strains in a HeLa cell invasion assay. We demonstrate that increasing the concentration of urea in the assay increased survival of the urease-positive but not urease-negative strains after 24 h, suggesting that urease does have a role in intracellular survival. Partial DNA sequence analysis of an 11.0 kb EcoRI DNA fragment encoding urease activity revealed an open reading frame containing 50%, 45%, 45%, and 41% homology to the UreA urease subunit protein of Klebsiella aerogenes, Proteus vulgaris, Helicobacter pylori and Proteus mirabilis respectively. We also show Bordetella pertussis to contain sequences homologous with a DNA probe containing the gene encoding UreA of B. bronchiseptica indicating the possible presence of cryptic urease genes in this species. PMID:8938644

  14. Analytical Verification of a PCR Assay for Identification of Bordetella avium

    Register, Karen B.; Yersin, Andrew G.


    Bordetella avium is the etiologic agent of turkey coryza or bordetellosis, a respiratory disease responsible for substantial economic losses to the turkey industry. At present, identification of this bacterium relies on isolation and biochemical testing. Although a PCR for the detection of B. avium was proposed a number of years ago (P. H. Savelkoul, L. E. de Groot, C. Boersma, I. Livey, C. J. Duggleby, B. A. van der Zeijst, and W. Gaastra, Microb. Pathog. 15:207-215, 1993), lack of analytica...

  15. Evaluation of Amplification Targets for the Specific Detection of Bordetella pertussis Using Real-Time Polymerase Chain Reaction

    Mohammad Rubayet Hasan


    Full Text Available BACKGROUND: Bordetella pertussis infections continue to be a major public health challenge in Canada. Polymerase chain reaction (PCR assays to detect B pertussis are typically based on the multicopy insertion sequence IS481, which offers high sensitivity but lacks species specificity.

  16. Bordetella bronchiseptica Pneumonia in an Infant and Genetic Comparison of Clinical Isolates with Veterinary Kennel Cough Vaccines

    An infant with recurrent episodes of respiratory failure was diagnosed with pertussis based on immunofluorescence testing, but culture revealed macrolide-resistant Bordetella bronchiseptica. Genetic analysis demonstrated that the child was not infected with a kennel cough vaccine strain, although th...

  17. Extracted protective antigen of Bordetella pertussis. I. Preparation and properties of the solubilized surface of components.

    Helting, T B; Blackkolb, F


    Bordetella pertussis microorganisms were treated with several extracting agents followed by ultracentrifugation to remove particulate matter. Analysis of the resulting supernatants by SDS gel electrophoresis showed one major component after simple salt extraction, and much more complex, although consistent pattern following detergent treatment. The yield of the solubilized protein in detergent extracts exceeded by far the values recorded for salt extracts. In order to prevent irreversible precipitation of the solubilized proteins upon removal of the denaturing agent, a novel procedure was developed. After extraction with urea-salt, the solubilized material was absorbed on a mineral carrier prior to the separation of the denaturing agent. The resulting absorbed vaccine was highly potent in the mouse-protection test, whereas the toxic reactions, elicited upon injection into experimental animals, were reduced in the comparison to the starting material. This diminished reactogenic potential was accompanied by the partial loss of the leukocytosis-promiting factor, whose activity was greatly diminished by urea-salt at alkaline pH-values. The procedure described may be applied to large-scale processing of Bordetella persussis microorganisms. Clinical trials now in progress should confirm or rebut the thesis that increased tolerability of the product, inferred from animal experiments, is reflected by fewer adverse reactions in humans. In the former case, the detergent extract vaccine may constitute a realistic alternative to conventional whole-cell vaccines against whooping-cough. PMID:6266198

  18. Comparative genomics of the classical Bordetella subspecies: the evolution and exchange of virulence-associated diversity amongst closely related pathogens

    Park Jihye


    Full Text Available Abstract Background The classical Bordetella subspecies are phylogenetically closely related, yet differ in some of the most interesting and important characteristics of pathogens, such as host range, virulence and persistence. The compelling picture from previous comparisons of the three sequenced genomes was of genome degradation, with substantial loss of genome content (up to 24% associated with adaptation to humans. Results For a more comprehensive picture of lineage evolution, we employed comparative genomic and phylogenomic analyses using seven additional diverse, newly sequenced Bordetella isolates. Genome-wide single nucleotide polymorphism (SNP analysis supports a reevaluation of the phylogenetic relationships between the classical Bordetella subspecies, and suggests a closer link between ovine and human B. parapertussis lineages than has been previously proposed. Comparative analyses of genome content revealed that only 50% of the pan-genome is conserved in all strains, reflecting substantial diversity of genome content in these closely related pathogens that may relate to their different host ranges, virulence and persistence characteristics. Strikingly, these analyses suggest possible horizontal gene transfer (HGT events in multiple loci encoding virulence factors, including O-antigen and pertussis toxin (Ptx. Segments of the pertussis toxin locus (ptx and its secretion system locus (ptl appear to have been acquired by the classical Bordetella subspecies and are divergent in different lineages, suggesting functional divergence in the classical Bordetellae. Conclusions Together, these observations, especially in key virulence factors, reveal that multiple mechanisms, such as point mutations, gain or loss of genes, as well as HGTs, contribute to the substantial phenotypic diversity of these versatile subspecies in various hosts.

  19. Produccion de suspensiones de bordetella pertussis por fermentación

    Algecira N.


    Full Text Available En este trabajo se estudió la producción de suspensión de Bordetella pertussis por fermentación para obtener el ingrediente activo de la vacuna contra tosferina. Se probaron diferentes medios de cultivo para el proceso, seleccionando el medio Stainer-Scholte adicionado con 3 g/L de casaminoacidos, el cual permite obtener altas concentraciones de células y suspensiones de buena calidad. Se estudió también la cinética de consumo de glutamato de sodio, producción de biomasa y evolución del pH. El crecimiento fue descrito por un modelo logístico. Se compara la tecnología de cultivo estacionario con el cultivo en fermentador presentándose esta última como la mejor alternativa de producción.

  20. Bordetella bronchiseptica Pneumonia in an Extremely-Low-Birth-Weight Neonate.

    Ting, Yuk Joseph; Ho, Pak-Leung; Wong, Kar-Yin


    Bordetella bronchiseptica, a gram-negative coccobacillus, is a common veterinary pathogen. In both domestic and wild animals, this bacterium causes respiratory infections including infectious tracheobronchitis in dogs and atrophic rhinitis in swine. Human infections are rare and have been documented in immunocompromised hosts. Here, we describe an extremely-low-birth-weight infant with B. bronchiseptica pneumonia. This is the first report that describes the microorganism's responsibility in causing nosocomial infection in a preterm neonate. He recovered uneventfully after a course of meropenem. It is possible that the bacteria colonize the respiratory tracts of our health care workers or parents who may have had contact with pets and then transmitted the bacterium to our patient. Follow-up until 21 months of age showed normal growth and development. He did not suffer from any significant residual respiratory disease. PMID:23705092

  1. Comparative Serological and Random Amplified Polymorphic DNA Typing for Bordetella avium Isolates in China

    Ping-Ping Yang, Rong-De Ma1, Xue Zhao and Rui-Liang Zhu*


    Full Text Available To study the similarity among Bordetella avium isolates in China, antigens and diagnostic antiserum of 22 B. avium isolates were prepared for serotyping, and a set of 20 commercially available primers was screened out to identify suitable primers for random amplified polymorphic DNA fingerprinting (RAPD analysis in this study. Twenty-two B. avium isolates were divided into two serovars (A and B based on their reaction in the plate-agglutination test. Four primers R1, R2, R4 and R10 resulted in informative fingerprints and were used to evaluate the B. avium isolates. Based on their RAPD patterns, a dendrogram allowed the separation of the B. avium isolates into six genetic similarity clusters. However, no direct correlation was observed between serotypes and RAPD typing among the isolates.

  2. Outer membrane protein OmpQ of Bordetella bronchiseptica is required for mature biofilm formation.

    Cattelan, Natalia; Villalba, María Inés; Parisi, Gustavo; Arnal, Laura; Serra, Diego Omar; Aguilar, Mario; Yantorno, Osvaldo


    Bordetella bronchiseptica, an aerobic Gram-negative bacterium, is capable of colonizing the respiratory tract of diverse animals and chronically persists inside the hosts by forming biofilm. Most known virulence factors in Bordetella species are regulated by the BvgAS two-component transduction system. The Bvg-activated proteins play a critical role during host infection. OmpQ is an outer membrane porin protein which is expressed under BvgAS control. Here, we studied the contribution of OmpQ to the biofilm formation process by B. bronchiseptica. We found that the lack of expression of OmpQ did not affect the growth kinetics and final biomass of B. bronchiseptica under planktonic growth conditions. The ΔompQ mutant strain displayed no differences in attachment level and in early steps of biofilm formation. However, deletion of the ompQ gene attenuated the ability of B. bronchiseptica to form a mature biofilm. Analysis of ompQ gene expression during the biofilm formation process by B. bronchiseptica showed a dynamic expression pattern, with an increase of biofilm culture at 48 h. Moreover, we demonstrated that the addition of serum anti-OmpQ had the potential to reduce the biofilm biomass formation in a dose-dependent manner. In conclusion, we showed for the first time, to the best of our knowledge, evidence of the contribution of OmpQ to a process of importance for B. bronchiseptica pathobiology. Our results indicate that OmpQ plays a role during the biofilm development process, particularly at later stages of development, and that this porin could be a potential target for strategies of biofilm formation inhibition. PMID:26673448

  3. Bordetella pertussis, the Causative Agent of Whooping Cough, Evolved from a Distinct, Human-Associated Lineage of B. bronchiseptica

    Diavatopoulos, Dimitri A; Cummings, Craig A.; Schouls, Leo M.; Brinig, Mary M.; Relman, David A.; Mooi, Frits R.


    Bordetella pertussis, B. bronchiseptica, B. parapertussishu, and B. parapertussisov are closely related respiratory pathogens that infect mammalian species. B. pertussis and B. parapertussishu are exclusively human pathogens and cause whooping cough, or pertussis, a disease that has resurged despite vaccination. Although it most often infects animals, infrequently B. bronchiseptica is isolated from humans, and these infections are thought to be zoonotic. B. pertussis and B. parapertussishu ar...

  4. Amidate Prodrugs of 9-[2-(Phosphonomethoxy)Ethyl]Adenine as Inhibitors of Adenylate Cyclase Toxin from Bordetella pertussis

    Šmídková, Markéta; Dvořáková, Alexandra; Tloušťová, Eva; Česnek, Michal; Janeba, Zlatko; Mertlíková-Kaiserová, Helena


    Roč. 58, č. 2 (2014), s. 664-671. ISSN 0066-4804 R&D Projects: GA MV VG20102015046 Grant ostatní: OPPC(XE) CZ.2.16/3.1.00/24016 Institutional support: RVO:61388963 Keywords : Bordetella pertussis * adenylate cyclase toxin * ACT * inhibitors * PMEA * amidate prodrugs Subject RIV: CC - Organic Chemistry Impact factor: 4.476, year: 2014

  5. Identification of residues essential for catalysis and binding of calmodulin in Bordetella pertussis adenylate cyclase by site-directed mutagenesis.

    Glaser, P; Elmaoglou-Lazaridou, A; Krin, E.; Ladant, D.; Bârzu, O; Danchin, A


    In order to identify molecular features of the calmodulin (CaM) activated adenylate cyclase of Bordetella pertussis, a truncated cya gene was fused after the 459th codon in frame with the alpha-lacZ' gene fragment and expressed in Escherichia coli. The recombinant, 604 residue long protein was purified to homogeneity by ion-exchange and affinity chromatography. The kinetic parameters of the recombinant protein are very similar to that of adenylate cyclase purified from B.pertussis culture sup...

  6. Bordetella pertussis filamentous hemagglutinin: evaluation as a protective antigen and colonization factor in a mouse respiratory infection model.

    Kimura, A; Mountzouros, K T; Relman, D.A.; Falkow, S; Cowell, J L


    Filamentous hemagglutinin (FHA) is a cell surface protein of Bordetella pertussis which functions as an adhesin for this organism. It is a component of many new acellular pertussis vaccines. The proposed role of FHA in immunity to pertussis is based on animal studies which have produced some conflicting results. To clarify this situation, we reexamined the protective activity of FHA in an adult mouse respiratory infection model. Four-week-old BALB/c mice were immunized with one or two doses o...

  7. A real-time PCR assay with improved specificity for detection and discrimination of all clinically relevant Bordetella species by the presence and distribution of three Insertion Sequence elements

    Ossewaarde Jacobus M


    Full Text Available Abstract Background In Dutch laboratories molecular detection of B. pertussis and B. parapertussis is commonly based on insertion sequences IS481 and IS1001, respectively. Both IS elements are more widely spread among Bordetella species. Both Bordetella holmesii, and B. bronchiseptica can harbour IS481. Also, IS1001 is found among B. bronchiseptica. IS481, and IS1001 based PCR thus lacks specificity when used for detection of specific Bordetella spp. Findings We designed a PCR based on IS1002, another IS element that is present among Bordetella species, and exploited it as a template in combination with PCR for IS481, and IS1001. In combining the PCRs for IS481, IS1001, and IS1002, and including an inhibition control, we were able to detect and discriminate all clinically relevant Bordetella species. Conclusions We developed an improved PCR method for specific detection of B. pertussis, B. parapertussis, B. holmesii, and B. bronchiseptica.

  8. Analysis of Bordetella pertussis clinical isolates circulating in European countries during the period 1998-2012.

    van Gent, M; Heuvelman, C J; van der Heide, H G; Hallander, H O; Advani, A; Guiso, N; Wirsing von Kőnig, C H; Vestrheim, D F; Dalby, T; Fry, N K; Pierard, D; Detemmerman, L; Zavadilova, J; Fabianova, K; Logan, C; Habington, A; Byrne, M; Lutyńska, A; Mosiej, E; Pelaz, C; Gröndahl-Yli-Hannuksela, K; Barkoff, A M; Mertsola, J; Economopoulou, A; He, Q; Mooi, F R


    Despite more than 50 years of vaccination, pertussis is still an endemic disease, with regular epidemic outbreaks. With the exception of Poland, European countries have replaced whole-cell vaccines (WCVs) by acellular vaccines (ACVs) in the 1990s. Worldwide, antigenic divergence in vaccine antigens has been found between vaccine strains and circulating strains. In this work, 466 Bordetella pertussis isolates collected in the period 1998-2012 from 13 European countries were characterised by multi-locus antigen sequence typing (MAST) of the pertussis toxin promoter (ptxP) and of the genes coding for proteins used in the ACVs: pertussis toxin (Ptx), pertactin (Prn), type 2 fimbriae (Fim2) and type 3 fimbriae (Fim3). Isolates were further characterised by fimbrial serotyping, multi-locus variable-number tandem repeat analysis (MLVA) and pulsed-field gel electrophoresis (PFGE). The results showed a very similar B. pertussis population for 12 countries using ACVs, while Poland, which uses a WCV, was quite distinct, suggesting that ACVs and WCVs select for different B. pertussis populations. This study forms a baseline for future studies on the effect of vaccination programmes on B. pertussis populations. PMID:25527446

  9. Rapid and sensitive detection of Bordetella bronchiseptica by loop-mediated isothermal amplification (LAMP

    Hui Zhang


    Full Text Available Bordetella bronchiseptica causes acute and chronic respiratory infections in diverse animal species and occasionally in humans. In this study, we described the establishment of a simple, sensitive and cost-efficient loop-mediated isothermal amplification (LAMP assay for the detection of B. bronchiseptica. A set of primers towards a 235 bp region within the flagellum gene of B. bronchiseptica was designed with online software.. The specificity of the LAMP assay was examined by using 6 porcine pathogens and 100 nasal swabs collected from healthy pigs and suspect infected pigs. The results indicated that positive reactions were confirmed for all B. bronchiseptica and no cross-reactivity was observed from other non-B. bronchiseptica. In sensitivity evaluations, the technique successfully detected a serial dilutions of extracted B. bronchiseptica DNA with a detection limit of 9 copies, which was 10 times more sensitive than that of PCR. Compared with conventional PCR, the higher sensitivity of LAMP method and no need for the complex instrumentation make this LAMP assay a promising alternative for the diagnosis of B. bronchiseptica in rural areas and developing countries where there lacks of complex laboratory services.

  10. Seroprevalence of Bordetella pertussis in the Mexican population: a cross-sectional study.

    Conde-Glez, C; Lazcano-Ponce, E; Rojas, R; DeAntonio, R; Romano-Mazzotti, L; Cervantes, Y; Ortega-Barría, E


    SUMMARY Serum samples collected during the National Health and Nutrition survey (ENSANUT 2006) were obtained from subjects aged 1-95 years (January-October 2010) and analysed to assess the seroprevalence of Bordetella pertussis (BP) in Mexico. Subjects' gender, age, geographical region and socioeconomic status were extracted from the survey and compiled into a subset database. A total of 3344 subjects (median age 29 years, range 1-95 years) were included in the analysis. Overall, BP seroprevalence was 47.4%. BP seroprevalence was significantly higher in males (53.4%, P = 0.0007) and highest in children (59.3%) decreasing with advancing age (P = 0.0008). BP seroprevalence was not significantly different between regions (P = 0.1918) and between subjects of socioeconomic status (P = 0.0808). Women, adolescents and young adults were identified as potential sources of infection to infants. Booster vaccination for adolescents and primary contacts (including mothers) for newborns and infants may provide an important public health intervention to reduce the disease burden. PMID:23734968

  11. Role of phosphoglucomutase of Bordetella bronchiseptica in lipopolysaccharide biosynthesis and virulence.

    West, N P; Jungnitz, H; Fitter, J T; McArthur, J D; Guzmán, C A; Walker, M J


    The phosphoglucomutase (PGM)-encoding gene of Bordetella bronchiseptica is required for lipopolysaccharide (LPS) biosynthesis. An insertion mutant of the wild-type B. bronchiseptica strain BB7865 which disrupted LPS biosynthesis was created and characterized (BB7865pgm). Genetic analysis of the mutated gene showed it shares high identity with PGM genes of various bacterial species and forms part of an operon which also encompasses the gene encoding phosphoglucose isomerase. Functional assays for PGM revealed that enzyme activity is expressed in both bvg-positive and bvg-negative strains of B. bronchiseptica and is substantially reduced in BB7865pgm. Complementation of the mutated PGM gene with that from BB7865 restored the wild-type condition for all phenotypes tested. The ability of the mutant BB7865pgm to survive within J774. A1 cells was significantly reduced at 2 h (40% reduction) and 24 h (56% reduction) postinfection. BB7865pgm was also significantly attenuated in its ability to survive in vivo following intranasal infection of mice, being effectively cleared from the lungs within 4 days, whereas the wild-type strain persisted at least 35 days. The activities of superoxide dismutase, urease, and acid phosphatase were unaffected in the PGM-deficient strain. In contrast, the inability to produce wild-type LPS resulted in a reduced bacterial resistance to oxidative stress and a higher susceptibility to the antimicrobial peptide cecropin P. PMID:10899872

  12. Membrane-Pore Forming Characteristics of the Bordetella pertussis CyaA-Hemolysin Domain

    Chattip Kurehong


    Full Text Available Previously, the 126-kDa Bordetella pertussis CyaA pore-forming/hemolysin (CyaA-Hly domain was shown to retain its hemolytic activity causing lysis of susceptible erythrocytes. Here, we have succeeded in producing, at large quantity and high purity, the His-tagged CyaA-Hly domain over-expressed in Escherichia coli as a soluble hemolytically-active form. Quantitative assays of hemolysis against sheep erythrocytes revealed that the purified CyaA-Hly domain could function cooperatively by forming an oligomeric pore in the target cell membrane with a Hill coefficient of ~3. When the CyaA-Hly toxin was incorporated into planar lipid bilayers (PLBs under symmetrical conditions at 1.0 M KCl, 10 mM HEPES buffer (pH 7.4, it produced a clearly resolved single channel with a maximum conductance of ~35 pS. PLB results also revealed that the CyaA-Hly induced channel was unidirectional and opened more frequently at higher negative membrane potentials. Altogether, our results first provide more insights into pore-forming characteristics of the CyaA-Hly domain as being the major pore-forming determinant of which the ability to induce such ion channels in receptor-free membranes could account for its cooperative hemolytic action on the target erythrocytes.

  13. Immunoproteomic Analysis ofBordetella bronchisepticaOuter Membrane Proteins and Identiifcation of New Immunogenic Proteins

    JI Quan-an


    Bordetella bronchiseptica is a Gram-negative pathogen that causes acute and chronic respiratory infection in a variety of animals. To identify useful antigen candidates for diagnosis and subunit vaccine ofB. bronchiseptica, immunoproteomic analysis was adopted to analyse outer membrane proteins of it. The outer membrane proteins extracted fromB. bronchiseptica were separated by two-dimensional gel electrophoresis and analyzed by Western blotting for their reactivity with the convalescent serum against two strains. Immunogenic proteins were identiifed by matrix-assisted laser desorption/ionization time of lfight-mass spectrometry (MALDI-TOF-MS), a total of 14 proteins are common immunoreactive proteins, of which 1 was known antigen and 13 were novel immunogenic proteins forB. bronchiseptica. Putative lipoprotein gene was cloned and recombinantly expressed. The recombinant protein induced high titer antibody, but showed low protective indices against challenges with HB (B. bronchiseptica strain isolated from a infected rabbit). The mortality of mice was 80% compared to 100% of positive controls. The identiifcation of these novel antigenic proteins is an important resource for further development of a new diagnostic test and vaccine for B. bronchiseptica.

  14. Comparative efficacy of intranasal and oral vaccines against Bordetella bronchiseptica in dogs.

    Ellis, J A; Gow, S P; Waldner, C L; Shields, S; Wappel, S; Bowers, A; Lacoste, S; Xu, Z; Ball, E


    In order to determine the comparative efficacy of vaccines administered intranasally or orally to protect puppies from disease subsequent to experimental infection with Bordetella bronchiseptica (Bb), a randomized controlled trial was performed using 48 approximately 8-week-old specific pathogen free, Bb naive Beagle puppies. Puppies were randomized into three groups and administered vaccines containing Bb intranasally or orally, or a placebo intranasally. Twenty-one days later, all dogs were challenge exposed via aerosol administration of Bb. Clinical signs, nasal bacterial shedding and immune responses were monitored for 28 days after challenge. Intranasally vaccinated puppies had significantly lower rates of coughing, nasal discharge, retching and sneezing (i.e. were less sick clinically) than control puppies. The distinction between the orally vaccinated puppies and the control puppies was less consistent. The orally vaccinated puppies had less coughing and less retching than the control puppies, but nasal discharge and sneezing did not differ from control animals. Orally vaccinated puppies had higher rates of coughing, nasal discharge, retching and sneezing than the intranasally vaccinated puppies. Although both intranasal and oral Bb vaccines stimulated immune responses associated with disease sparing following Bb infection, the intranasal route of delivery conferred superior clinical outcomes. The observed difference in clinical efficacy suggests the need to question the rationale for the use of currently available orally administered Bb vaccines. PMID:27256028

  15. Modulation of the NF-kappaB pathway by Bordetella pertussis filamentous hemagglutinin.

    Tzvia Abramson

    Full Text Available BACKGROUND: Filamentous hemagglutinin (FHA is a cell-associated and secreted adhesin produced by Bordetella pertussis with pro-apoptotic and pro-inflammatory activity in host cells. Given the importance of the NF-kappaB transcription factor family in these host cell responses, we examined the effect of FHA on NF-kappaB activation in macrophages and bronchial epithelial cells, both of which are relevant cell types during natural infection. METHODOLOGY/PRINCIPAL FINDINGS: Exposure to FHA of primary human monocytes and transformed U-937 macrophages, but not BEAS-2B epithelial cells, resulted in early activation of the NF-kappaB pathway, as manifested by the degradation of cytosolic IkappaB alpha, by NF-kappaB DNA binding, and by the subsequent secretion of NF-kappaB-regulated inflammatory cytokines. However, exposure of macrophages and human monocytes to FHA for two hours or more resulted in the accumulation of cytosolic IkappaB alpha, and the failure of TNF-alpha to activate NF-kappaB. Proteasome activity was attenuated following exposure of cells to FHA for 2 hours, as was the nuclear translocation of RelA in BEAS-2B cells. CONCLUSIONS: These results reveal a complex temporal dynamic, and suggest that despite short term effects to the contrary, longer exposures of host cells to this secreted adhesin may block NF-kappaB activation, and perhaps lead to a compromised immune response to this bacterial pathogen.

  16. Crystal violet staining of Bordetella bronchiseptica colonies for differentiation of phase-I strains from variant strains in degraded phases.

    Ishikawa, H.; Isayama, Y


    After 2 days of growth on Brain heart infusion agar (BHIA) at 38 degrees C, phase-I colonies and degraded-phase colonies of Bordetella bronchiseptica could be differentiated by their ability to take up crystal violet (CV). Phase-I colonies in X mode, but not colonies in degraded phases (phases II, III, and rough) bound CV. Phenotypically-altered C-mode colonies (grown at 32 degrees C or lower temperatures) also lacked this ability. CV staining offers an easy method for the recognition of diff...

  17. Molecular evolution of the two-component system BvgAS involved in virulence regulation in Bordetella.

    Julien Herrou

    Full Text Available The whooping cough agent Bordetella pertussis is closely related to Bordetella bronchiseptica, which is responsible for chronic respiratory infections in various mammals and is occasionally found in humans, and to Bordetella parapertussis, one lineage of which causes mild whooping cough in humans and the other ovine respiratory infections. All three species produce similar sets of virulence factors that are co-regulated by the two-component system BvgAS. We characterized the molecular diversity of BvgAS in Bordetella by sequencing the two genes from a large number of diverse isolates. The response regulator BvgA is virtually invariant, indicating strong functional constraints. In contrast, the multi-domain sensor kinase BvgS has evolved into two different types. The pertussis type is found in B. pertussis and in a lineage of essentially human-associated B. bronchiseptica, while the bronchiseptica type is associated with the majority of B. bronchiseptica and both ovine and human B. parapertussis. BvgS is monomorphic in B. pertussis, suggesting optimal adaptation or a recent population bottleneck. The degree of diversity of the bronchiseptica type BvgS is markedly different between domains, indicating distinct evolutionary pressures. Thus, absolute conservation of the putative solute-binding cavities of the two periplasmic Venus Fly Trap (VFT domains suggests that common signals are perceived in all three species, while the external surfaces of these domains vary more extensively. Co-evolution of the surfaces of the two VFT domains in each type and domain swapping experiments indicate that signal transduction in the periplasmic region may be type-specific. The two distinct evolutionary solutions for BvgS confirm that B. pertussis has emerged from a specific B. bronchiseptica lineage. The invariant regions of BvgS point to essential parts for its molecular mechanism, while the variable regions may indicate adaptations to different lifestyles. The

  18. Filamentous hemagglutinin has a major role in mediating adherence of Bordetella pertussis to human WiDr cells.

    Urisu, A; Cowell, J L; Manclark, C R


    [35S]methionine-labeled Bordetella pertussis adhered to monolayers of WiDr cells, an epitheliumlike cell line from a human intestinal carcinoma. Adherence was proportional to the density of the WiDr cells and to the concentration of B. pertussis in the assay. Adherence of virulent phase I strains Tohama phase I, 114, and BP338 was much greater than adherence of avirulent strains Tohama phase III and 423 phase IV. Mutants deficient in the production of the filamentous hemagglutinin (FHA) were ...

  19. Enhanced Ex Vivo Stimulation of Mycobacterium tuberculosis-Specific T Cells in Human Immunodeficiency Virus-Infected Persons via Antigen Delivery by the Bordetella pertusis Adenylate Cyclase Vector

    Connell, T. G.; Shey, M. S.; Seldon, R.; Rangaka, M. X.; van Cutsem, G.; Šimšová, Marcela; Marčeková, Zuzana; Šebo, Peter; Curtis, N.; Diwakar, L.; Meintjes, G. A.; Leclerc, C.; Wilkinson, R. J.; Wilkinson, K. A.


    Roč. 14, č. 7 (2007), s. 847-854. ISSN 1556-6811 R&D Projects: GA MŠk 2B06161 Institutional research plan: CEZ:AV0Z50200510 Keywords : mycobacterium tuberculosis * bordetella pertusis * human immunodeficiency virus Subject RIV: EE - Microbiology, Virology Impact factor: 1.995, year: 2007

  20. Expression of bvg-repressed genes in Bordetella pertussis is controlled by RisA through a novel c-di-GMP signaling pathway

    The BvgAS two component system of Bordetella pertussis controls virulence factor expression. In addition, BvgAS controls expression of the bvg-repressed genes through the action of the repressor, BvgR. The transcription factor RisA is inhibited by BvgR, and when BvgR is not expressed RisA induces th...

  1. Cyclic di-GMP regulation of the bvg-repressed genes and the orphan response regulator RisA in Bordetella pertussis

    Expression of Bordetella pertussis virulence factors is activated by the BvgAS two-component system. Under modulating growth conditions BvgAS indirectly represses another set of genes through the action of BvgR, a bvg-activated protein. BvgR blocks activation of the response regulator RisA which is ...

  2. Bisamidate Prodrugs of 2-Substituted 9-[2-(Phosphonomethoxy)ethyl]adenine (PMEA, adefovir) as Selective Inhibitors of Adenylate Cyclase Toxin from Bordetella pertussis

    Česnek, Michal; Jansa, Petr; Šmídková, Markéta; Mertlíková-Kaiserová, Helena; Dračínský, Martin; Brust, T. F.; Pávek, P.; Trejtnar, F.; Watts, V. J.; Janeba, Zlatko


    Roč. 10, č. 8 (2015), s. 1351-1364. ISSN 1860-7179 R&D Projects: GA MV VG20102015046 Institutional support: RVO:61388963 Keywords : adenylate cyclase toxin * bisamidates * Bordetella pertussis * nucleosides * phosphonates Subject RIV: CC - Organic Chemistry Impact factor: 2.968, year: 2014

  3. Acylation of Lysine 860 Allows Tight Binding and Cytotoxicity of Bordetella Adenylate Cyclase on CD1 1b-Expressing Cells

    Mašín, Jiří; Basler, Marek; Knapp, O.; El-Azami-El-Idrissi, M.; Maier, E.; Konopásek, I.; Benz, R.; Leclerc, C.; Šebo, Peter


    Roč. 44, - (2005), s. 12766-12759. ISSN 0006-2960 R&D Projects: GA AV ČR IAA5020406; GA MŠk 1M0506 Institutional research plan: CEZ:AV0Z50200510 Keywords : lysine 860 * bordetella Subject RIV: EE - Microbiology, Virology Impact factor: 3.848, year: 2005

  4. Research on pharmacological mechanism of the treatment of Asthma by oral Bordetella pertussis

    CHI Shen; SUN Yun; ZHANG Bao-yuan


    Objective To examine the effect of oral Bordetella pertussis on the asthma mice sensitized by ovalbumin (OVA), and explore the possible mechanism. Methods Culture the B. pertussis in Bordet-Gengou agar containing 25 % rabbit blood. Collect the bacteria and inactive them at 80 ℃ for 30 min to get whole killed B. pertussis. 32 BALB/C mice were randomly divided into control group, model-control group, model group and treatment group. The mice were sensitized and challenged with OVA to establish asthma model. Asthma mice in treatment group were orally administrated with B. pertussis 7 days before sensitization. The mice in control group and model-control group were challenged with saline. After 24 hours of last challenge, bronchoaveolar lavage fluid (BALF) and peripheral blood were collected. The total cells and eosinophils were counted in BALF. Results Compared with the control group (2.03±0.42, 0.33±0.82)× 105 mL-1 and model-control group (2.16±0.48,0.16±0.41)×105 mL-1, the total cells (10.13±1.33) ×105mL-1 and eosinophils (11.83±4.573)×105 mL-1 in BALF were more in asthma mice (P<0.01). The number of total cells (5.50±1.55)×105 mL-1 and eosinophils(0.66±0.82)×105 mL-1 in BALF were reduced in asthma mice treated with B. pertussis compared with asthma mice(P<0.01 ). Conclusions Oral B. pertussis can inhabit airway inflammation of asthma mice and has the potential of treating asthma.

  5. Cooperative phenomena in binding and activation of Bordetella pertussis adenylate cyclase by calmodulin.

    Bouhss, A; Krin, E; Munier, H; Gilles, A M; Danchin, A; Glaser, P; Bârzu, O


    The catalytic domain of Bordetella pertussis adenylate cyclase located within the first 400 amino acids of the protein can be cleaved by trypsin in two subdomains (T25 and T18) corresponding to ATP-(T25) and calmodulin (CaM)-(T18) binding sites. Reassociation of subdomains by CaM is a cooperative process, which is a unique case among CaM-activated enzymes. To understand better the molecular basis of this phenomenon, we used several approaches such as partial deletions of the adenylate cyclase gene, isolation of peptides of various size, and site-directed mutagenesis experiments. We found that a stretch of 72 amino acid residues overlapping the carboxyl terminus of T25 and the amino terminus of T18 accounts for 90% of the binding energy of adenylate cyclase-CaM complex. The hydrophobic "side" of the helical region situated around Trp242 plays a major role in the interaction of adenylate cyclase with CaM, whereas basic residues that alternate with acidic residues in bacterial enzyme play a much less important role. The amino-terminal half of the catalytic domain of adenylate cyclase contributes only 10% to the binding energy of CaM, whereas the last 130 amino acid residues are not at all involved in binding. However, these segments of adenylate cyclase might affect protein/protein interaction and catalysis by propagating conformational changes to the CaM-binding sequence which is located in the middle of the catalytic domain of bacterial enzyme. PMID:8420945

  6. Antimicrobial Susceptibility of Bordetella bronchiseptica Isolates from Swine and Companion Animals and Detection of Resistance Genes.

    Sandra Prüller

    Full Text Available Bordetella bronchiseptica causes infections of the respiratory tract in swine and other mammals and is a precursor for secondary infections with Pasteurella multocida. Treatment of B. bronchiseptica infections is conducted primarily with antimicrobial agents. Therefore it is essential to get an overview of the susceptibility status of these bacteria. The aim of this study was to comparatively analyse broth microdilution susceptibility testing according to CLSI recommendations with an incubation time of 16 to 20 hours and a longer incubation time of 24 hours, as recently proposed to obtain more homogenous MICs. Susceptibility testing against a panel of 22 antimicrobial agents and two fixed combinations was performed with 107 porcine isolates from different farms and regions in Germany and 43 isolates obtained from companion animals in Germany and other European countries. Isolates with increased MICs were investigated by PCR assays for the presence of resistance genes. For ampicillin, all 107 porcine isolates were classified as resistant, whereas only a single isolate was resistant to florfenicol. All isolates obtained from companion animals showed elevated MICs for β-lactam antibiotics and demonstrated an overall low susceptibility to cephalosporines. Extension of the incubation time resulted in 1-2 dilution steps higher MIC50 values of porcine isolates for seven antimicrobial agents tested, while isolates from companion animals exhibited twofold higher MIC50/90 values only for tetracycline and cefotaxime. For three antimicrobial agents, lower MIC50 and MIC90 values were detected for both, porcine and companion animal isolates. Among the 150 isolates tested, the resistance genes blaBOR-1 (n = 147, blaOXA-2, (n = 4, strA and strB (n = 17, sul1 (n = 10, sul2 (n = 73, dfrA7 (n = 3 and tet(A (n = 8 were detected and a plasmid localisation was identified for several of the resistance genes.

  7. Cilia-associated bacteria in fatal Bordetella bronchiseptica pneumonia of dogs and cats.

    Taha-Abdelaziz, Khaled; Bassel, Laura L; Harness, Melanie L; Clark, Mary Ellen; Register, Karen B; Caswell, Jeff L


    Bordetella bronchiseptica frequently causes nonfatal tracheobronchitis, but its role in fatal pneumonia is less recognized. Our study evaluated histologic identification of cilia-associated bacteria as a method for diagnosis of B. bronchiseptica pneumonia. Cases of fatal bronchopneumonia were studied retrospectively, excluding neonates and cases of aspiration pneumonia, minor lung lesions, or autolysis. The study population comprised 36 canine and 31 feline cases of bronchopneumonia. B. bronchiseptica was identified in 8 of 36 canine and 14 of 31 feline cases based on immunohistochemistry (IHC) using serum from a rabbit hyperimmunized with pertactin, PCR testing (Fla2/Fla12), and/or bacterial culture data when available. Of these, IHC was positive in 4 canine and 7 feline cases, PCR was positive in 8 canine and 14 feline cases, and B. bronchiseptica was isolated in 2 of 5 canine and 3 of 9 feline cases tested. Examination of histologic sections stained with hematoxylin and eosin revealed bronchial cilia-associated bacteria in 4 of 36 canine and 5 of 31 feline cases; these were all positive by IHC and PCR. The presence of cilia-associated bacteria had been noted in the pathology report for only 2 of these 9 cases. Thus, the presence of cilia-associated bacteria seems frequently overlooked by pathologists, but is a diagnostically significant feature of B. bronchiseptica pneumonia. A specific diagnosis of B. bronchiseptica pneumonia is important because it suggests primary or opportunistic bacterial pneumonia rather than aspiration pneumonia, and because of the risk of animal-to-animal transmission of B. bronchiseptica, the availability of vaccines for disease prevention, and the potential zoonotic risk to immunocompromised pet owners. PMID:27178716

  8. Expresión episomal de toxina de pertussis genéticamente inactivada en Bordetella pertussis

    Ernesto Marcos


    Full Text Available Bordetella pertussis es una bacteria Gram negativa, la cual constituye el agente etiologico de la tos ferina. La enfermedad se desencadena por el efecto conjunto de una serie de factores de virulencia expresados por la bacteria, los cuales se encuentran regulados por el sistema bvg. Uno de los factores de virulencia mas importantes es la toxina de pertussis, razon por la cual, se emplea de forma inactivada como el componente principal de las vacunas acelulares contra la enfermedad. La toxina de pertussis posee una estructura del tipo A-B compuesta por seis polipeptidos codificados en un operon unico. El polipeptido S1 constituye la subunidad enzimaticamente activa, la cual cataliza la transferencia de ADP-ribosa del NAD a la subunidad ALPHA de las proteinas G en celulas eucariotas, lo cual genera una serie de efectos biologicos dentro de los que se incluye: sensibilizacion a histamina, incremento de la secrecion de insulina y efectos inmunoestimuladores e inmunosupresores. El presente trabajo describe los procedimientos realizados para la obtencion de cepas de Bordetella pertussis productoras de elevadas concentraciones de toxina pertusica atenuada geneticamente. Para esto, se realizaron las sustituciones aminoacidicas Arg9 por Lys y Glu129 por Gly de la subunidad S1. El operon de la toxina de pertussis mutada se clono en un vector de amplio rango de hospedero bajo la regulacion de un promotor de expresion temprana (fhaB. Los clones obtenidos pudieran ser empleados como sistemas de expresion para produccion de vacunas acelulares en Cuba.

  9. Analysis of Bordetella pertussis pertactin and pertussis toxin types from Queensland, Australia, 1999–2003

    Slack Andrew T


    Full Text Available Abstract Background In Australia two acellular Bordetella pertussis vaccines have replaced the use of a whole cell vaccine. Both of the licensed acellular vaccines contain the following three components; pertussis toxoid, pertussis filamentous haemagglutinin and the 69 kDa pertactin adhesin. One vaccine also contains pertussis fimbriae 2 and 3. Various researchers have postulated that herd immunity due to high levels of pertussis vaccination might be influencing the makeup of endemic B. pertussis populations by selective pressure for strains possessing variants of these genes, in particular the pertactin gene type. Some publications have suggested that B. pertussis variants may be contributing to a reduced efficacy of the existing vaccines and a concomitant re-emergence of pertussis within vaccinated populations. This study was conducted to survey the pertactin and pertussis toxin subunit 1 types from B. pertussis isolates in Queensland, Australia following the introduction of acellular vaccines. Methods Forty-six B. pertussis isolates recovered from Queensland patients between 1999 and 2003 were examined by both DNA sequencing and LightCycler™ real time PCR to determine their pertactin and pertussis toxin subunit 1 genotypes. Results Pertactin typing showed that 38 isolates possessed the prn1 allele, 3 possessed the prn2 allele and 5 possessed the prn3 allele. All forty-six isolates possessed the pertussis toxin ptxS1A genotype. Amongst the circulating B. pertussis population in Queensland, 82.5% of the recovered clinical isolates therefore possessed the prn1/ptxS1A genotype. Conclusion The results of this study compared to historical research on Queensland isolates suggest that B. pertussis pertactin and pertussis toxin variants are not becoming more prevalent in Queensland since the introduction of the acellular vaccines. Current prevalences of pertactin variants are significantly different to that described in a number of other countries

  10. Infección por Bordetella pertussis: Una causa emergente de tos prolongada en adolescentes y adultos Bordetella pertussis infection: An emerging cause of prolonged cough illness in adolescents and adults



    Full Text Available La tos convulsiva o coqueluche está siendo reconocida cada vez con mayor frecuencia como causa de tos prolongada en adolescentes y adultos. La vacunación sistemática de la población pediátrica ha determinado un cambio en el perfl epidemiológico de la enfermedad, aumentando su prevalencia en la población adulta. Se presenta el caso clínico de una paciente de 45 años, fumadora, enfermera de unidad de hemodiálisis, que consulta por malestar general y tos seca de seis semanas de evolución. La radiografía de tórax era normal y la inmunofuorescencia directa de hisopado nasofaríngeo fue positiva para Bordetella pertussis. A propósito de este caso clínico, revisamos las principales causas de tos crónica: asma bronquial, enfermedad rinosinusal y refujo gastroesofágico; el cuadro clínico, evaluación diagnóstica y tratamiento de la infección por B. pertussis en población adulta.Whooping cough is increasingly recognized as a cause of prolonged cough illness in adolescents and adults. Systematic vaccination has changed its epidemiology, with the majority of cases now primarily affecting adolescents and adults. A 45-year-old female, active smoker, nurse, who works in a dialysis service, presented with a 6-week history of bothersome cough and malaise. Thorax x-ray was normal and direct immunofuorescence of nasopharyngeal swab was positive for Bordetella pertussis. This case illustrates pertussis infection in adulthood. We review the main causes of chronic cough in adults: asthma, chronic rhinosinusitis and gastroesophageal refux; the clinical features, prevalence, diagnostic tools, and management of adult patients with B. pertussis infection to increase awareness of this highly contagious disease.

  11. A new assay for invasion of HeLa 229 cells by Bordetella pertussis: effects of inhibitors, phenotypic modulation, and genetic alterations.

    Lee, C. K.; Roberts, A. L.; Finn, T M; Knapp, S; Mekalanos, J J


    Invasion and intracellular survival of Bordetella pertussis in HeLa 229 cells was studied by a new assay that utilizes polymyxin B instead of gentamicin to rapidly kill extracellular organisms. Invasion measured by this assay was time and temperature dependent and was inhibited by the microfilament drug cytochalasin D. The invasion process was also dependent on a functional vir locus (also known as bvg), the positive regulator of virulence gene expression in B. pertussis. Four spontaneous Vir...

  12. Murine antibody response to oral infection with live aroA recombinant Salmonella dublin vaccine strains expressing filamentous hemagglutinin antigen from Bordetella pertussis.

    Molina, N C; Parker, C D


    Two plasmids which express either nearly intact or truncated filamentous hemagglutinin (FHA) from Bordetella pertussis and which are marked with a tetracycline resistance (Tcr) gene were transformed into Salmonella dublin SL1438, an aroA deletion mutant intended for use as an attenuated oral vaccine against salmonellosis. These S. dublin recombinants, when fed to mice, induced serum immunoglobulin, immunoglobulin M (IgM), and sometimes IgA antibody responses to FHA and S. dublin. In addition,...

  13. [Serological evaluation of Bordetella pertussis infection in adults with prolonged cough].

    Sönmez, Cemile; Çöplü, Nilay; Gözalan, Ayşegül; Yılmaz, Ülkü; Bilekli, Selen; Demirci, Nilgün Yılmaz; Biber, Çiğdem; Erdoğan, Yurdanur; Esen, Berrin; Çöplü, Lütfi


    Pertussis is a vaccine-preventable disease that is transmitted from infected to susceptible individuals by respiratory route. Bordetella pertussis infection may occur at any age as neither vaccine nor natural infection induced immunity lasts life-long. This study was planned to demonstrate the serological evidence of infection among adults, to raise awareness among clinicians and to provide data for the development of strategies to protect vulnerable infants. A total of 538 patients (345 female, 193 male) ages between 18-87 years who had a complain of prolonged cough for more than two weeks were included in the study. Anti-pertussis toxin (PT) IgG and anti-filamentous hemagglutinin (FH) IgG levels from single serum samples were measured by an in-house ELISA test which was standardized and shown to be efficient previously. Anti-PT IgG antibody levels of ≥ 100 EU/ml were considered as acute/recent infection with B.pertussis. In our study, 9.7% (52/538) of the patients had high levels of anti-PT IgG (≥ 100 EU/ml) and among those patients 43 (43/52; 82.7%) also had high (≥ 100 EU/ml) anti-FHA IgG levels. There were no statistically significant differences in terms of age, gender, education level, DPT (diphtheria-pertussis-tetanus) vaccination history, smoking history or average daily cigarette consumption (p> 0.05) between the cases with high antibody levels (n= 52). When the symptoms and the presence of cases with high antibody levels were evaluated, it was detected that no one parameter was significantly different from others, except that 24.1% of the cases with inspiratory whooping had high anti-PT levels. There was also no statistically significant difference between high anti-PT levels ≥ 100 EU/ml and the patients with risk factors [smoking (21/200; 10.5%), presence of disease that cause chronic cough and/or drug usage (19/171; %11.1), and whole factors which cause chronic cough (32/306; %10.5)] and without risk factors (p= 0.581; p= 0.357; p= 0

  14. Laboratory-based surveillance of pertussis using multitarget real-time PCR in Japan: evidence for Bordetella pertussis infection in preteens and teens

    K. Kamachi


    Full Text Available Between January 2013 and December 2014, we conducted laboratory-based surveillance of pertussis using multitarget real-time PCR, which discriminates among Bordetella pertussis, Bordetella parapertussis, Bordetella holmesii and Mycoplasma pneumoniae. Of 355 patients clinically diagnosed with pertussis in Japan, B. pertussis, B. parapertussis and M. pneumoniae were detected in 26% (n = 94, 1.1% (n = 4 and 0.6% (n = 2, respectively, whereas B. holmesii was not detected. It was confirmed that B. parapertussis and M. pneumoniae are also responsible for causing pertussis-like illness. The positive rates for B. pertussis ranged from 16% to 49%, depending on age. Infants aged ≤ 3 months had the highest rate (49%, and children aged 1 to 4 years had the lowest rate (16%, p < 0.01 vs. infants aged ≤ 3 months. Persons aged 10 to 14 and 15 to 19 years also showed high positive rates (29% each; the positive rates were not statistically significant compared with that of infants aged ≤ 3 months (p ≥ 0.06. Our observations indicate that similar to infants, preteens and teens are at high risk of B. pertussis infection.

  15. Mutation in the β-hairpin of the Bordetella pertussis adenylate cyclase toxin modulates N-lobe conformation in calmodulin

    Springer, Tzvia I.; Goebel, Erich; Hariraju, Dinesh [Department of Microbiology, Miami University, Oxford, OH 45056 (United States); Finley, Natosha L., E-mail: [Department of Microbiology, Miami University, Oxford, OH 45056 (United States); Cell, Molecular, and Structural Biology Program, Miami University, Oxford, OH 45056 (United States)


    Highlights: • Bordetella pertussis adenylate cyclase toxin modulates bi-lobal structure of CaM. • The structure and stability of the complex rely on intermolecular associations. • A novel mode of CaM-dependent activation of the adenylate cyclase toxin is proposed. - Abstract: Bordetella pertussis, causative agent of whooping cough, produces an adenylate cyclase toxin (CyaA) that is an important virulence factor. In the host cell, the adenylate cyclase domain of CyaA (CyaA-ACD) is activated upon association with calmodulin (CaM), an EF-hand protein comprised of N- and C-lobes (N-CaM and C-CaM, respectively) connected by a flexible tether. Maximal CyaA-ACD activation is achieved through its binding to both lobes of intact CaM, but the structural mechanisms remain unclear. No high-resolution structure of the intact CaM/CyaA-ACD complex is available, but crystal structures of isolated C-CaM bound to CyaA-ACD shed light on the molecular mechanism by which this lobe activates the toxin. Previous studies using molecular modeling, biochemical, and biophysical experiments demonstrate that CyaA-ACD’s β-hairpin participates in site-specific interactions with N-CaM. In this study, we utilize nuclear magnetic resonance (NMR) spectroscopy to probe the molecular association between intact CaM and CyaA-ACD. Our results indicate binding of CyaA-ACD to CaM induces large conformational perturbations mapping to C-CaM, while substantially smaller structural changes are localized primarily to helices I, II, and IV, and the metal-binding sites in N-CaM. Site-specific mutations in CyaA-ACD’s β-hairpin structurally modulate N-CaM, resulting in conformational perturbations in metal binding sites I and II, while no significant structural modifications are observed in C-CaM. Moreover, dynamic light scattering (DLS) analysis reveals that mutation of the β-hairpin results in a decreased hydrodynamic radius (R{sub h}) and reduced thermal stability in the mutant complex. Taken

  16. Investigations into the emergence of pertactin-deficient Bordetella pertussis isolates in six European countries, 1996 to 2012.

    Zeddeman, A; van Gent, M; Heuvelman, C J; van der Heide, H G; Bart, M J; Advani, A; Hallander, H O; Wirsing von Konig, C H; Riffelman, M; Storsaeter, J; Vestrheim, D F; Dalby, T; Krogfelt, K A; Fry, N K; Barkoff, A M; Mertsola, J; He, Q; Mooi, F


    Pathogen adaptation has been proposed to contribute to the resurgence of pertussis. A striking recent example is the emergence of isolates deficient in the vaccine component pertactin (Prn). This study explores the emergence of such Prn-deficient isolates in six European countries. During 2007 to 2009, 0/83 isolates from the Netherlands, 0/18 from the United Kingdom, 0/17 Finland, 0/23 Denmark, 4/99 Sweden and 5/20 from Norway of the isolates collected were Prn-deficient. In the Netherlands and Sweden, respectively 4/146 and 1/8 were observed in a later period (2010–12). The Prn-deficient isolates were genetically diverse and different mutations were found to inactivate the prn gene. These are indications that Prn-deficiency is subject to positive selective pressure. We hypothesise that the switch from whole cell to acellular pertussis vaccines has affected the balance between ‘costs and benefits’ of Prn production by Bordetella pertussis to the extent that isolates that do not produce Prn are able to expand. The absence of Prn-deficient isolates in some countries may point to ways to prevent or delay the spread of Prn-deficient strains. In order to substantiate this hypothesis, trends in the European B. pertussis population should be monitored continuously. PMID:25166348

  17. Plasmacytoid dendritic cell-derived IFNα modulates Th17 differentiation during early Bordetella pertussis infection in mice.

    Wu, V; Smith, A A; You, H; Nguyen, T A; Ferguson, R; Taylor, M; Park, J E; Llontop, P; Youngman, K R; Abramson, T


    Whooping cough is a highly contagious respiratory disease caused by Bordetella pertussis (B. pertussis). T helper 17 (Th17) cells have a central role in the resolution of the infection. Emerging studies document that type I interferons (IFNs) suppress Th17 differentiation and interleukin (IL)-17 responses in models of infection and chronic inflammation. As plasmacytoid dendritic cells (pDCs) are a major source of type I IFNs, we hypothesize that during B. pertussis infection in mice, pDC-derived IFNα inhibits a rapid increase in Th17 cells. We found that IFNα-secreting pDCs appear in the lungs during the early stages of infection, while a robust rise of Th17 cells in the lungs is detected at 15 days post-infection or later. The presence of IFNα led to reduced Th17 differentiation and proliferation in vitro. Furthermore, in vivo blocking of IFNα produced by pDCs during infection with B. pertussis infection resulted in early increase of Th17 frequency, inflammation, and reduced bacterial loads in the airways of infected mice. Taken together, the experiments reported here describe an inhibitory role for pDCs and pDC-derived IFNα in modulating Th17 responses during the early stages of B. pertussis infection, which may explain the prolonged nature of whooping cough. PMID:26462419

  18. Homologs of the LapD-LapG c-di-GMP Effector System Control Biofilm Formation by Bordetella bronchiseptica.

    Nicolás Ambrosis

    Full Text Available Biofilm formation is important for infection by many pathogens. Bordetella bronchiseptica causes respiratory tract infections in mammals and forms biofilm structures in nasal epithelium of infected mice. We previously demonstrated that cyclic di-GMP is involved in biofilm formation in B. bronchiseptica. In the present work, based on their previously reported function in Pseudomonas fluorescens, we identified three genes in the B. bronchiseptica genome likely involved in c-di-GMP-dependent biofilm formation: brtA, lapD and lapG. Genetic analysis confirmed a role for BrtA, LapD and LapG in biofilm formation using microtiter plate assays, as well as scanning electron and fluorescent microscopy to analyze the phenotypes of mutants lacking these proteins. In vitro and in vivo studies showed that the protease LapG of B. bronchiseptica cleaves the N-terminal domain of BrtA, as well as the LapA protein of P. fluorescens, indicating functional conservation between these species. Furthermore, while BrtA and LapG appear to have little or no impact on colonization in a mouse model of infection, a B. bronchiseptica strain lacking the LapG protease has a significantly higher rate of inducing a severe disease outcome compared to the wild type. These findings support a role for c-di-GMP acting through BrtA/LapD/LapG to modulate biofilm formation, as well as impact pathogenesis, by B. bronchiseptica.

  19. Homologs of the LapD-LapG c-di-GMP Effector System Control Biofilm Formation by Bordetella bronchiseptica

    Ambrosis, Nicolás; Boyd, Chelsea D.; O´Toole, George A.; Fernández, Julieta; Sisti, Federico


    Biofilm formation is important for infection by many pathogens. Bordetella bronchiseptica causes respiratory tract infections in mammals and forms biofilm structures in nasal epithelium of infected mice. We previously demonstrated that cyclic di-GMP is involved in biofilm formation in B. bronchiseptica. In the present work, based on their previously reported function in Pseudomonas fluorescens, we identified three genes in the B. bronchiseptica genome likely involved in c-di-GMP-dependent biofilm formation: brtA, lapD and lapG. Genetic analysis confirmed a role for BrtA, LapD and LapG in biofilm formation using microtiter plate assays, as well as scanning electron and fluorescent microscopy to analyze the phenotypes of mutants lacking these proteins. In vitro and in vivo studies showed that the protease LapG of B. bronchiseptica cleaves the N-terminal domain of BrtA, as well as the LapA protein of P. fluorescens, indicating functional conservation between these species. Furthermore, while BrtA and LapG appear to have little or no impact on colonization in a mouse model of infection, a B. bronchiseptica strain lacking the LapG protease has a significantly higher rate of inducing a severe disease outcome compared to the wild type. These findings support a role for c-di-GMP acting through BrtA/LapD/LapG to modulate biofilm formation, as well as impact pathogenesis, by B. bronchiseptica PMID:27380521

  20. A Type VI secretion system encoding locus is required for Bordetella bronchiseptica immunomodulation and persistence in vivo.

    Laura S Weyrich

    Full Text Available Type VI Secretion Systems (T6SSs have been identified in numerous gram-negative pathogens, but the lack of a natural host infection model has limited analysis of T6SS contributions to infection and pathogenesis. Here, we describe disruption of a gene within locus encoding a putative T6SS in Bordetella bronchiseptica strain RB50, a respiratory pathogen that circulates in a broad range of mammals, including humans, domestic animals, and mice. The 26 gene locus encoding the B. bronchiseptica T6SS contains apparent orthologs to all known core genes and possesses thirteen novel genes. By generating an in frame deletion of clpV, which encodes a putative ATPase required for some T6SS-dependent protein secretion, we observe that ClpV contributes to in vitro macrophage cytotoxicity while inducing several eukaryotic proteins associated with apoptosis. Additionally, ClpV is required for induction of IL-1β, IL-6, IL-17, and IL-10 production in J774 macrophages infected with RB50. During infections in wild type mice, we determined that ClpV contributes to altered cytokine production, increased pathology, delayed lower respiratory tract clearance, and long term nasal cavity persistence. Together, these results reveal a natural host infection system in which to interrogate T6SS contributions to immunomodulation and pathogenesis.

  1. Expression of the Putative Siderophore Receptor Gene bfrZ Is Controlled by the Extracytoplasmic-Function Sigma Factor BupI in Bordetella bronchiseptica

    Pradel, Elizabeth; Locht, Camille


    A new gene from Bordetella bronchiseptica, bfrZ encoding a putative siderophore receptor, was identified in a Fur-repressor titration assay. A bfrZ null mutant was constructed by allelic exchange. The protein profile of this mutant is similar to that of the wild-type parent strain. The BfrZ−-BfrZ+ isogenic pair was tested for utilization of 132 different siderophores as iron sources. None of these iron sources acted as a ligand for BfrZ. Translational bfrZ::phoA and transcriptional bfrZ::lacZ...

  2. Characterization of the N-terminal domain of BteA: a Bordetella type III secreted cytotoxic effector.

    Chen Guttman

    Full Text Available BteA, a 69-kDa cytotoxic protein, is a type III secretion system (T3SS effector in the classical Bordetella, the etiological agents of pertussis and related mammalian respiratory diseases. Currently there is limited information regarding the structure of BteA or its subdomains, and no insight as to the identity of its eukaryotic partners(s and their modes of interaction with BteA. The mechanisms that lead to BteA dependent cell death also remain elusive. The N-terminal domain of BteA is multifunctional, acting as a docking platform for its cognate chaperone (BtcA in the bacterium, and targeting the protein to lipid raft microdomains within the eukaryotic host cell. In this study we describe the biochemical and biophysical characteristics of this domain (BteA287 and determine its architecture. We characterize BteA287 as being a soluble and highly stable domain which is rich in alpha helical content. Nuclear magnetic resonance (NMR experiments combined with size exclusion and analytical ultracentrifugation measurements confirm these observations and reveal BteA287 to be monomeric in nature with a tendency to oligomerize at concentrations above 200 µM. Furthermore, diffusion-NMR demonstrated that the first 31 residues of BteA287 are responsible for the apparent aggregation behavior of BteA287. Light scattering analyses and small angle X-ray scattering experiments reveal a prolate ellipsoidal bi-pyramidal dumb-bell shape. Thus, our biophysical characterization is a first step towards structure determination of the BteA N-terminal domain.

  3. Immuno-enhancement of Taishan Pinus massoniana pollen polysaccharides on recombinant Bordetella avium ompA expressed in Pichia pastoris.

    Liu, Liping; Yu, Cuilian; Wang, Chuanwen; Shao, Mingxu; Yan, Zhengui; Jiang, Xiaodong; Chi, Shanshan; Wang, Zhen; Wei, Kai; Zhu, Ruiliang


    Bordetellosis, caused by Bordetella avium, continues to be an economic problem in the poultry industry of China. Vaccines with good protective ability are lacking. Thus, developing a novel vaccine against the B. avium infection is crucial. Here, we constructed a recombinant Pichia pastoris transformant capable of expressing the outer membrane protein A (ompA) of B. avium to prepare the recombinant ompA subunit vaccine and then evaluated its immune effects. To further investigate the immunomodulation effects of Taishan Pinus massoniana pollen polysaccharides (TPPPS) on this subunit vaccine, three concentrations (20, 40, and 60 mg/mL) of TPPPS were used as the adjuvants of the ompA subunit vaccine respectively. The conventional Freund's incomplete adjuvant served as the control of TPPPS. Chickens in different groups were separately vaccinated with these vaccines thrice. During the monitoring period, serum antibody titers, concentrations of serum IL-4, percentages of CD4(+) and CD8(+) T-lymphocytes in the peripheral blood, lymphocyte transformation rate, and protection rate were detected. Results showed that the pure ompA vaccine induced the production of anti-ompA antibody, the secretion of IL-4, the increase of CD4(+) T-lymphocytes counts and lymphocyte transformation rate in the peripheral blood. Moreover, the pure ompA vaccine provided a protection rate of 71.67% after the B. avium challenge. Notably, TPPPS adjuvant vaccines induced higher levels of immune responses than the pure ompA vaccine, and 60 mg/mL TPPPS adjuvant vaccine showed optimal immune effects and had a 91.67% protection rate. Our findings indicated that this recombinant B. avium ompA subunit vaccine combined with TPPPS had high immunostimulatory potential. Results provided a new perspective for B. avium subunit vaccine research. PMID:26975477

  4. Purification and assay of cell-invasive form of calmodulin-sensitive adenylyl cyclase from Bordetella pertussis

    An invasive form of the CaM-sensitive adenylyl cyclase from Bordetella pertussis can be isolated from bacterial culture supernatants. This isolation is achieved through the use of QAE-Sephadex anion-exchange chromatography. It has been demonstrated that the addition of exogenous Ca2+ to the anion-exchange gradient buffers will affect elution from the column and will thereby affect the isolation of invasive adenylyl cyclase. This is probably due to a Ca2(+)-dependent interaction of the catalytic subunit with another component in the culture supernatant. Two peaks of adenylyl cyclase activity are obtained. The Pk1 adenylyl cyclase preparation is able to cause significant increases in intracellular cAMP levels in animal cells. This increase occurs rapidly and in a dose-dependent manner in both N1E-115 mouse neuroblastoma cells and human erythrocytes. The Pk2 adenylyl cyclase has catalytic activity but is not cell invasive. This material can serve, therefore, as a control to ensure that the cAMP which is measured is, indeed, intracellular. A second control is to add exogenous CaM to the Pk1 adenylyl cyclase preparation. The 45-kDa catalytic subunit-CaM complex is not cell invasive. Although the mechanism for membrane translocation of the adenylyl cyclase is unknown, there is evidence that the adenylyl cyclase enters animal cells by a mechanism distinct from receptor-mediated endocytosis. Calmodulin-sensitive adenylyl cyclase activity can be removed from preparations of the adenylyl cyclase that have been subjected to SDS-polyacrylamide gel electrophoresis. This property of the enzyme has enabled purification of the catalytic subunit to apparent homogeneity. The purified catalytic subunit from culture supernatants has a predicted molecular weight of 45,000. This polypeptide interacts directly with Ca2+ and this interaction may be important for its invasion into animal cells

  5. ε-Caprolactam Utilization by Proteus sp. and Bordetella sp. Isolated From Solid Waste Dumpsites in Lagos State, Nigeria, First Report.

    Sanuth, Hassan Adeyemi; Yadav, Amit; Fagade, Obasola Ezekiel; Shouche, Yogesh


    The ε-caprolactam is the monomer of the synthetic non-degradable nylon-6 and often found as nonreactive component of nylon-6 manufacturing waste effluent. Environmental consequences of its toxicity to natural habitats and humans pose a global public concern. Soil samples were collected from three designated solid waste dumpsites, namely, Abule-Egba, Olusosun and Isheri-Igando in Lagos State, Nigeria. Sixteen bacteria isolated from these samples were found to utilize the ε-caprolactam as a sole source of carbon and nitrogen at concentration of ≤20 g l(-1). The isolates were characterized using their 16S rRNA gene sequence and showed similarity with Pseudomonas sp., Proteus sp., Providencia sp., Corynebacterium sp., Lysinibacillus sp., Leucobacter sp., Alcaligenes sp. and Bordetella sp. Their optimal growth conditions were found to be at temperature range of 30 to 35 °C and pH range of 7.0-7.5. High Performance liquid chromatography analysis of the ε-caprolactam from supernatant of growth medium revealed that these isolates have potential to remove 31.6-95.7 % of ε-caprolactam. To the best of our knowledge, this study is first to report the ability of Proteus sp. and Bordetella sp. for ε-caprolactam utilization. PMID:24426112

  6. Genome Structural Diversity among 31 Bordetella pertussis Isolates from Two Recent U.S. Whooping Cough Statewide Epidemics

    Bowden, Katherine E.; Weigand, Michael R.; Peng, Yanhui; Cassiday, Pamela K.; Sammons, Scott; Knipe, Kristen; Rowe, Lori A.; Loparev, Vladimir; Sheth, Mili; Weening, Keeley; Tondella, M. Lucia


    ABSTRACT During 2010 and 2012, California and Vermont, respectively, experienced statewide epidemics of pertussis with differences seen in the demographic affected, case clinical presentation, and molecular epidemiology of the circulating strains. To overcome limitations of the current molecular typing methods for pertussis, we utilized whole-genome sequencing to gain a broader understanding of how current circulating strains are causing large epidemics. Through the use of combined next-generation sequencing technologies, this study compared de novo, single-contig genome assemblies from 31 out of 33 Bordetella pertussis isolates collected during two separate pertussis statewide epidemics and 2 resequenced vaccine strains. Final genome architecture assemblies were verified with whole-genome optical mapping. Sixteen distinct genome rearrangement profiles were observed in epidemic isolate genomes, all of which were distinct from the genome structures of the two resequenced vaccine strains. These rearrangements appear to be mediated by repetitive sequence elements, such as high-copy-number mobile genetic elements and rRNA operons. Additionally, novel and previously identified single nucleotide polymorphisms were detected in 10 virulence-related genes in the epidemic isolates. Whole-genome variation analysis identified state-specific variants, and coding regions bearing nonsynonymous mutations were classified into functional annotated orthologous groups. Comprehensive studies on whole genomes are needed to understand the resurgence of pertussis and develop novel tools to better characterize the molecular epidemiology of evolving B. pertussis populations. IMPORTANCE Pertussis, or whooping cough, is the most poorly controlled vaccine-preventable bacterial disease in the United States, which has experienced a resurgence for more than a decade. Once viewed as a monomorphic pathogen, B. pertussis strains circulating during epidemics exhibit diversity visible on a genome

  7. A Bordetella pertussis proteoliposome induces protection in mice without affecting the immunogenicity of diphtheria and tetanus toxoids in a trivalent formulation.

    Castillo, Sonsire Fernández; Chovel, Mario Landys; Hernández, Niurka Gutiérrez; González, Lorena Corcho; Blanco, Amaya; Hernández, Daily Serrano; Medina, Mildrey Fariñas; Tito, Maydelis Álvarez; Quiñoy, José Luis Pérez


    In this study, a formulation of Bordetella pertussis proteoliposome (PLBp), diphtheria, and tetanus toxoids and alum (DT-PLBp) was evaluated as a trivalent vaccine candidate in BALB/c mice. Vaccine-induced protection was estimated using the intranasal challenge for pertussis and enzyme-linked immunosorbent assay fvto assess serological responses for diphtheria or tetanus. Both, diphtheria-tetanus-whole cell pertussis (DTP) and diphtheria-tetanus vaccines (DT) were used as controls. Animals immunized with DT-PLBp, PLBp alone, and DTP showed total reduction of CFU in lungs 7 days after intranasal challenge. Likewise, formulations DT-PLBp, DTP, and DT elicited antibody levels ≥2 IU/mL against tetanus and diphtheria, considered protective when neutralization tests are used. Overall, results showed that combination of PLBp with tetanus and diphtheria toxoids did not affect the immunogenicity of each antigen alone. PMID:27489808

  8. Bordetella protein toxins

    Mašín, Jiří; Šebo, Peter; Locht, C.

    New York : Elsevier, Academic Press, 2006, s. 291-309. ISBN 978-0-12-088445-2 R&D Projects: GA AV ČR IAA5020406; GA MŠk 1M0506 Institutional research plan: CEZ:AV0Z50200510 Keywords : pertussis toxin * adenylate cyclase toxin * dermonecrotic toxin Subject RIV: EE - Microbiology, Virology

  9. Cloning and sequencing of a gene encoding a 21-kilodalton outer membrane protein from Bordetella avium and expression of the gene in Salmonella typhimurium.

    Gentry-Weeks, C R; Hultsch, A L; Kelly, S M; Keith, J M; Curtiss, R


    Three gene libraries of Bordetella avium 197 DNA were prepared in Escherichia coli LE392 by using the cosmid vectors pCP13 and pYA2329, a derivative of pCP13 specifying spectinomycin resistance. The cosmid libraries were screened with convalescent-phase anti-B. avium turkey sera and polyclonal rabbit antisera against B. avium 197 outer membrane proteins. One E. coli recombinant clone produced a 56-kDa protein which reacted with convalescent-phase serum from a turkey infected with B. avium 197. In addition, five E. coli recombinant clones were identified which produced B. avium outer membrane proteins with molecular masses of 21, 38, 40, 43, and 48 kDa. At least one of these E. coli clones, which encoded the 21-kDa protein, reacted with both convalescent-phase turkey sera and antibody against B. avium 197 outer membrane proteins. The gene for the 21-kDa outer membrane protein was localized by Tn5seq1 mutagenesis, and the nucleotide sequence was determined by dideoxy sequencing. DNA sequence analysis of the 21-kDa protein revealed an open reading frame of 582 bases that resulted in a predicted protein of 194 amino acids. Comparison of the predicted amino acid sequence of the gene encoding the 21-kDa outer membrane protein with protein sequences in the National Biomedical Research Foundation protein sequence data base indicated significant homology to the OmpA proteins of Shigella dysenteriae, Enterobacter aerogenes, E. coli, and Salmonella typhimurium and to Neisseria gonorrhoeae outer membrane protein III, Haemophilus influenzae protein P6, and Pseudomonas aeruginosa porin protein F. The gene (ompA) encoding the B. avium 21-kDa protein hybridized with 4.1-kb DNA fragments from EcoRI-digested, chromosomal DNA of Bordetella pertussis and Bordetella bronchiseptica and with 6.0- and 3.2-kb DNA fragments from EcoRI-digested, chromosomal DNA of B. avium and B. avium-like DNA, respectively. A 6.75-kb DNA fragment encoding the B. avium 21-kDa protein was subcloned into the

  10. A direct pyrophosphatase-coupled assay provides new insights into the activation of the secreted adenylate cyclase from Bordetella pertussis by calmodulin.

    Lawrence, Anthony J; Coote, John G; Kazi, Yasmin F; Lawrence, Paul D; MacDonald-Fyall, Julia; Orr, Barbara M; Parton, Roger; Riehle, Mathis; Sinclair, James; Young, John; Price, Nicholas C


    Continuous recording of the activity of recombinant adenylate cyclase (CyaA) of Bordetella pertussis (EC ) by conductimetric determination of enzyme-coupled pyrophosphate cleavage has enabled us to define a number of novel features of the activation of this enzyme by calmodulin and establish conditions under which valid activation data can be obtained. Activation either in the presence or absence of calcium is characterized by a concentration-dependent lag phase. The rate of formation and breakdown of the activated complex can be determined from an analysis of the lag phase kinetics and is in good agreement with thermodynamic data obtained by measuring the dependence of activation on calmodulin concentration, which show that calcium increases k(on) by about 30-fold. The rate of breakdown of the activated complex, formed either in the presence or absence of calcium, has been determined by dilution experiments and has been shown to be independent of the presence of calcium. The coupled assay is established as a rapid, convenient and safe method which should be readily applicable to the continuous assays of most other enzymes that catalyze reactions in which inorganic pyrophosphate is liberated. PMID:11934879

  11. Cloning of Bordetella bronchiseptica urease genes and analysis of colonization by a urease-negative mutant strain in a guinea-pig model.

    Monack, D M; Falkow, S


    The genes encoding urease were cloned from Bordetella bronchiseptica and the 5.2 kb of DNA essential for expression analysed in a T7 RNA polymerase transcription-translation system. At least four polypeptides with predicted molecular weights of 69,000, 26,000, 12,200 and 11,000 were found. Partial DNA sequence of the gene encoding the 69,000 Da polypeptide revealed high amino acid identity to the alpha-subunit of Proteus mirabilis urease, UreC and jack bean urease. A stable, unmarked deletion was constructed in this gene to create a urease-negative mutant of B. bronchiseptica. To assess colonization in a guinea-pig model, the urease-negative strain was inoculated with the urease-positive parental strain in a mixed infection. The urease-negative strain out competed the urease-positive strain in the trachea, lungs and caecum. We demonstrate that urease is not essential for B. bronchiseptica colonization of the guinea-pig respiratory and digestive tracts. PMID:7968532

  12. A versatile, non genetically modified organism (GMO)-based strategy for controlling low-producer mutants in Bordetella pertussis cultures using antigenic modulation.

    Goffin, Philippe; Slock, Thomas; Smessaert, Vincent; De Rop, Philippe; Dehottay, Philippe


    The uncontrolled presence of non-producer mutants negatively affects bioprocesses. In Bordetella pertussis cultures, avirulent mutants emerge spontaneously and accumulate. We characterized the dynamics of accumulation using high-throughput growth assays and competition experiments between virulent and avirulent (bvg(-) ) isolates. A fitness advantage of bvg(-) cells was identified as the main driver for bvg(-) accumulation under conditions of high virulence factor production. Conversely, under conditions that reduce their expression (antigenic modulation), bvg(-) takeover could be avoided. A control strategy was derived, which consists in applying modulating conditions whenever virulence factor production is not required. It has a wide range of applications, from routine laboratory operations to vaccine manufacturing, where pertussis toxin yields were increased 1.4-fold by performing early pre-culture steps in modulating conditions. Because it only requires subtle modifications of the culture medium and does not involve genetic modifications, this strategy is applicable to any B. pertussis isolate, and should facilitate regulatory acceptance of process changes for vaccine production. Strategies based on the same concept, could be derived for other industrially relevant micro-organisms. This study illustrates how a sound scientific understanding of physiological principles can be turned into a practical application for the bioprocess industry, in alignment with Quality by Design principles. PMID:26014907

  13. Crystallization and preliminary X-ray diffraction analysis of two extracytoplasmic solute receptors of the DctP family from Bordetella pertussis

    Sample preparation, crystallization and preliminary X-ray analysis are reported for two B. pertussis extracytoplasmic solute receptors. DctP6 and DctP7 are two Bordetella pertussis proteins which belong to the extracytoplasmic solute receptors (ESR) superfamily. ESRs are involved in the transport of substrates from the periplasm to the cytosol of Gram-negative bacteria. DctP6 and DctP7 have been crystallized and diffraction data were collected using a synchrotron-radiation source. DctP6 crystallized in space group P41212, with unit-cell parameters a = 108.39, b = 108.39, c = 63.09 Å, while selenomethionyl-derivatized DctP7 crystallized in space group P212121, with unit-cell parameters a = 64.87, b = 149.83, c = 170.65 Å. The three-dimensional structure of DctP7 will be determined by single-wavelength anomalous diffraction, while the DctP6 structure will be solved by molecular-replacement methods

  14. Bordetella adenylate cyclase toxin mobilizes its beta2 integrin receptor into lipid rafts to accomplish translocation across target cell membrane in two steps.

    Ladislav Bumba


    Full Text Available Bordetella adenylate cyclase toxin (CyaA binds the alpha(Mbeta(2 integrin (CD11b/CD18, Mac-1, or CR3 of myeloid phagocytes and delivers into their cytosol an adenylate cyclase (AC enzyme that converts ATP into the key signaling molecule cAMP. We show that penetration of the AC domain across cell membrane proceeds in two steps. It starts by membrane insertion of a toxin 'translocation intermediate', which can be 'locked' in the membrane by the 3D1 antibody blocking AC domain translocation. Insertion of the 'intermediate' permeabilizes cells for influx of extracellular calcium ions and thus activates calpain-mediated cleavage of the talin tether. Recruitment of the integrin-CyaA complex into lipid rafts follows and the cholesterol-rich lipid environment promotes translocation of the AC domain across cell membrane. AC translocation into cells was inhibited upon raft disruption by cholesterol depletion, or when CyaA mobilization into rafts was blocked by inhibition of talin processing. Furthermore, CyaA mutants unable to mobilize calcium into cells failed to relocate into lipid rafts, and failed to translocate the AC domain across cell membrane, unless rescued by Ca(2+ influx promoted in trans by ionomycin or another CyaA protein. Hence, by mobilizing calcium ions into phagocytes, the 'translocation intermediate' promotes toxin piggybacking on integrin into lipid rafts and enables AC enzyme delivery into host cytosol.

  15. Conservación por congelación de Bordetella pertussis y Corynebacterium diphtheriae, empleados en la producción de vacunas para uso humano

    Yilian Plasencia,


    Full Text Available En el presente estudio se evaluó el método de congelación a –70ºC para la preservación de Bordetella pertussis y Corynebacterium diphtheriae. Para verificar el sustento de los cultivos se realizó un adecuado control de calidad, que incluyó comprobación de pureza, viabilidad y estabilidad de las propiedades de interés. En este trabajo se probaron diferentes formulaciones. Se seleccionó la que arrojó los mejores resultados y se realizó un estudio de mantenimiento de las características evaluadas durante el tiempo. Para medir determinados parámetros se realizaron procesos a escala industrial, empleándose para esto un biorreactor Chemap de 35 L. Se tomaron como referencia los valores obtenidos por las cepas conservadas por liofilización. De esta forma se buscaron alternativas y soluciones a problemas presentados en su conservación. Los resultados obtenidos sugieren la posible inclusión en el Programa de Mantenimiento establecido.

  16. Association between Pneumocystis spp. and co-infections with Bordetella bronchiseptica, Mycoplasma hyopneumoniae and Pasteurella multocida in Austrian pigs with pneumonia.

    Kureljušić, B; Weissenbacher-Lang, C; Nedorost, N; Stixenberger, D; Weissenböck, H


    In this retrospective study, 218 pig lung tissue samples were analyzed to examine a possible association between Pneumocystis spp. using in situ hybridization, Bordetella bronchiseptica (B.b.) using immunohistochemistry (IHC), Mycoplasma hyopneumoniae (M.h.) by quantitative PCR, and Pasteurella multocida (P.m.; IHC). Compared to the bacterial agents (B.b., 5%; M.h., 30%; P.m., 23%), Pneumocystis occurred with a higher prevalence (51%). Co-infections with two or three pathogens were present in 28% of the examined cases. Those of Pneumocystis and M.h. were most commonly seen, followed by Pneumocystis and P.m. and M.h. and P.m. Histologically, interstitial pneumonia was found in both the Pneumocystis positive lungs and lungs with a mild M.h. infection. The B.b. and P.m. positive lungs were mainly associated with suppurative bronchopneumonia and severe M.h. cases with fibrinous or fibrino-haemorrhagic pneumonia. In suckling piglets, the number of samples positive for Pneumocystis predominated, whereas samples from fattening pigs were mainly positive for bacteria or Pneumocystis and bacteria. PMID:26654847

  17. Differences in Purinergic Amplification of Osmotic Cell Lysis by the Pore-Forming RTX Toxins Bordetella pertussis CyaA and Actinobacillus pleuropneumoniae ApxIA: the Role of Pore Size

    Mašín, Jiří; Fišer, Radovan; Linhartová, Irena; Osička, Radim; Bumba, Ladislav; Hewlett, E. L.; Benz, R.; Šebo, Peter


    Roč. 81, č. 12 (2013), s. 4571-4582. ISSN 0019-9567 R&D Projects: GA ČR GAP302/12/0460; GA ČR(CZ) GAP302/11/0580; GA ČR(CZ) GAP207/11/0717; GA AV ČR IAA500200914; GA ČR GA13-14547S Institutional support: RVO:61388971 Keywords : Bordetella pertussis * Actinobacillus pleuropneumoniae * E-coli Subject RIV: EE - Microbiology, Virology Impact factor: 4.156, year: 2013

  18. Tosse convulsa em Portugal: análise retrospetiva de casos clínicos suspeitos de infeção por Bordetella pertussis no período 2010-2014

    Santos, Maria Augusta; Pereira, Brigida; Furtado, Cristina


    Objetivo: Analisar retrospetivamente os resultados laboratoriais dos casos clínicos suspeitos de tosse convulsa enviados ao Laboratório Nacional de Referência de Bordetella pertussis do INSA no Porto para confirmação laboratorial no período 2010-2014, tendo como finalidade alertar não só para a necessidade de conhecer-se melhor a real incidência da tosse convulsa nos grupos etários com idade superior a 13 anos, bem como de caracterizar geneticamente as estirpes circulantes em Portuga...

  19. cAMP signalling of Bordetella adenylate cyclase toxin through the SHP-1 phosphatase activates the BimEL-Bax pro-apoptotic cascade in phagocytes.

    Ahmad, Jawid Nazir; Cerny, Ondrej; Linhartova, Irena; Masin, Jiri; Osicka, Radim; Sebo, Peter


    The adenylate cyclase toxin-hemolysin (CyaA, ACT or AC-Hly) plays a key role in virulence of Bordetella pertussis. CyaA penetrates myeloid cells expressing the complement receptor 3 (αM β2 integrin CD11b/CD18) and subverts bactericidal capacities of neutrophils and macrophages by catalysing unregulated conversion of cytosolic ATP to the key signalling molecule adenosine 3',5'-cyclic monophosphate (cAMP). We show that the signalling of CyaA-produced cAMP hijacks, by an as yet unknown mechanism, the activity of the tyrosine phosphatase SHP-1 and activates the pro-apoptotic BimEL-Bax cascade. Mitochondrial hyperpolarization occurred in human THP-1 macrophages within 10 min of exposure to low CyaA concentrations (e.g. 20 ng ml(-1) ) and was accompanied by accumulation of BimEL and association of the pro-apoptotic factor Bax with mitochondria. BimEL accumulation required cAMP/protein kinase A signalling, depended on SHP-1 activity and was selectively inhibited upon small interfering RNA knockdown of SHP-1 but not of the SHP-2 phosphatase. Moreover, signalling of CyaA-produced cAMP inhibited the AKT/protein kinase B pro-survival cascade, enhancing activity of the FoxO3a transcription factor and inducing Bim transcription. Synergy of FoxO3a activation with SHP-1 hijacking thus enables the toxin to rapidly trigger a persistent accumulation of BimEL, thereby activating the pro-apoptotic programme of macrophages and subverting the innate immunity of the host. PMID:26334669

  20. Pesquisa de antigenos aglutinantes "major" 1, 2 e 3 em cepas de Bordetella pertussis, isoladas de crianças com coqueluche atendidas no Hospital de Isolamento Emílio Ribas de São Paulo, Brasil Determination of 1, 2 and 3 major antigens in Bordetella pertussis strains isolated from Brazilian children with whooping-cough

    Sebastião Timo Iaria


    Full Text Available Em 30 cepas de Bordetella pertussis isoladas de crianças com coqueluche, atendidas no Hospital de Isolamento Emílio Ribas de São Paulo, foram pesquisados os antígenos aglutinantes ''major" 1, 2 e 3. Levando-se em conta a presença combinada dos três antígenos, as provas de soro-aglutinação rápida em lamina revelaram que 25 (83,3% cepas possuiam os fatores 1, 2 e 3, enquanto que 3 (10,0% e 2 (6,7% foram positivas, somente, para 1, 2 e 1, 3, respectivamente. Os resultados foram discutidos, considerando-se a importância deste antígeno no preparo de vacinas.The presence of major antigens, 1, 2 and 3 were determined in 30 strains of B. pertussis isolated from children with whooping-cough hospitalized at the Hospital Emílio Ribas, São Paulo Brazil. The method used was the slide-agglutination test. Tests showed that 25(83.3% of strains were positives for factors 1, 2 and 3. Factores 1 and 3 alone were present in 3 (10% of strains and 1 and 2 alone in 2 (6.7%.

  1. Performance of transport and selective media for swine Bordetella bronchiseptica recovery and it comparison to polymerase chain reaction detection Desempenho de meios de transporte e seletivo na recuperação de Bordetella bronchiseptica de suínos e sua comparação à detecção pela reação em cadeia pela polimerase

    Tania Alen Coutinho


    Full Text Available Three comparative assays were performed seeking to improve the sensitivity of the diagnosis of Bordetella bronchiseptica infection analyzing swine nasal swabs. An initial assay compared the recovery of B. bronchiseptica from swabs simultaneously inoculated with B. bronchiseptica and some interfering bacteria, immersed into three transport formulations (Amies with charcoal, trypticase soy broth and phosphate buffer according to Soerensen supplemented with 5% of bovine fetal serum and submitted to different temperatures (10ºC and 27ºC and periods of incubation (24, 72 and 120 hours. A subsequent assay compared three selective media (MacConkey agar, modified selective medium G20G and a ceftiofur medium for their recovery capabilities from clinical specimens. One last assay compared the polymerase chain reaction to the three selective media. In the first assay, the recovery of B. bronchiseptica from transport systems was better at 27ºC and the three formulations had good performances at this temperature, but the collection of qualitative and quantitative analysis indicated the advantage of Amies medium for nasal swabs transportation. The second assay indicated that MacConkey agar and modified G20G had similar results and were superior to the ceftiofur medium. In the final assay, polymerase chain reaction presented superior capability of B. bronchiseptica detection to culture procedures.Três ensaios comparativos foram feitos com o objetivo de aperfeiçoar a sensibilidade do diagnóstico da infecção pela Bordetella bronchiseptica a partir de suabes nasais de leitões. O experimento inicial comparou a recuperação de B. bronchiseptica a partir de suabes, simultaneamente inoculados com B. bronchiseptica e algumas bactérias interferentes, imersos em três formulações para transporte (meio Amies com carvão, caldo tripticaseína de soja e tampão de fosfatos segundo Soerensen suplementado com 5% de soro fetal bovino e submetidos a diferentes

  2. Ca2+ influx and tyrosine kinases trigger Bordetella adenylate cyclase toxin (ACT endocytosis. Cell physiology and expression of the CD11b/CD18 integrin major determinants of the entry route.

    Kepa B Uribe

    Full Text Available Humans infected with Bordetella pertussis, the whooping cough bacterium, show evidences of impaired host defenses. This pathogenic bacterium produces a unique adenylate cyclase toxin (ACT which enters human phagocytes and catalyzes the unregulated formation of cAMP, hampering important bactericidal functions of these immune cells that eventually cause cell death by apoptosis and/or necrosis. Additionally, ACT permeabilizes cells through pore formation in the target cell membrane. Recently, we demonstrated that ACT is internalised into macrophages together with other membrane components, such as the integrin CD11b/CD18 (CR3, its receptor in these immune cells, and GM1. The goal of this study was to determine whether ACT uptake is restricted to receptor-bearing macrophages or on the contrary may also take place into cells devoid of receptor and gain more insights on the signalling involved. Here, we show that ACT is rapidly eliminated from the cell membrane of either CR3-positive as negative cells, though through different entry routes, which depends in part, on the target cell physiology and characteristics. ACT-induced Ca(2+ influx and activation of non-receptor Tyr kinases into the target cell appear to be common master denominators in the different endocytic strategies activated by this toxin. Very importantly, we show that, upon incubation with ACT, target cells are capable of repairing the cell membrane, which suggests the mounting of an anti-toxin cell repair-response, very likely involving the toxin elimination from the cell surface.

  3. 兔支气管败血波氏杆菌DNT蛋白的分段表达及其免疫保护性研究%The expression and immunoprotection of the truncated dermonecrotic toxin of rabbit Bordetella bronchiseptica

    赵宁; 王芳; 恽时锋; 范志宇; 胡波


    Dermonecrotic toxin (DNT) is identified as one of the main virulence factor of Bordetella bronchiseptica. To study the immunoprotection of the binding and catalytic regions of DNT, the N-terminal 1,612 bp (DNT1) and the C-terminal 973 bp (DNT3) fragments of the DNT were cloned into the pET-28a(+) and expressed in E. Coli BL21(DE3), respectively. The expression of recombinant protein DNT1 and DNT3 were identified by SDS-PAGE and western blot analysis. High level antibody against DNT was induced in the mice immunized with the recombinant proteins, and all the mice in immunized groups were survived when challenged intraperitoneally with virulent B. Bronchiseptic strain BJL0504 at 2 dosages of LD50, while the mice mortility were 60% in control group. Furthermore, the bacteria in lung was eliminated completely from the immunned mice post challenging intranasally with B. Bronchiseptic (1.07 × 108 cfu/mL) at 9 days, on the contrary, the bactera counts in mice lung were up to 1.2 × 105 cfu in control group. These results shown that the DNT1 and DNT3 had strong immunoprotection against B. Bronchiseptic infection and could be used as a subunit vaccine against B. Bronchiseptic.%皮肤坏死毒素(DNT)是支气管败血波氏杆菌(Bb)的主要毒力因子之一.为研究DNT结合位点和催化位点的免疫保护性,本研究分别将其核苷酸序列N-端的1 612bp片段(DNT1)和C-端的973bp片段(DNT3)克隆到原核表达载体pET-28a(+)中,经IPTG诱导后在E.coli BL21 (DE3)中获得表达.SDS-PAGE和western blot检测表明重组蛋白DNT1、DNT3均具有良好的反应原性.在主动免疫保护试验中,重组蛋白免疫组小鼠均能够产生较高的DNT抗体水平;当使用2 LD50的Bb强毒株BJL0504进行腹腔攻毒后,其存活率为100%,而阴性对照组则为40%;当使用1.07×108 cfu/mL Bb强毒株BJL0504进行滴鼻攻毒后第9d时,重组蛋白免疫组小鼠已基本清除肺脏内的Bb菌,而阴性对照组小鼠

  4. Bordetella adenylate cyclase toxin: a swift saboteur of host defense

    Vojtová, Jana; Kamanová, Jana; Šebo, Peter


    Roč. 9, - (2006), s. 1-7. ISSN 1369-5274 R&D Projects: GA AV ČR IAA5020406; GA MŠk 1M0506 Institutional research plan: CEZ:AV0Z50200510 Keywords : cyaa * scanning electron microscopy * cyclase toxin Subject RIV: EE - Microbiology, Virology Impact factor: 7.445, year: 2006


    Karataev, G I; Sinyashina, L N; Medkova, A Yu; Semin, E G


    A growth of pertussis morbidity is observed in many countries of the world against the background of mass vaccindtion. Forms of the disease course have changed. Atypical forms of pertussis occur predominately in adolescents and adults. Asymptomatic carriage of the causative agent has been established. Infection of infants with. BordetelIa pertussis bacteria in more than 90% of cases occurs from parents and relatives. A prolonged persistence of the causative agent has been identified. Morbidity increase in developed countries is associated with the use of acellular vaccines, that do not protect from the infection, but reduce severity of the disease. A change of genotypes of the circulating bacteria strains is observed ubiquitously. Formation of a persistent form of B. pertussis is possible due to a reversible integration of IS-elements into bvgAS operon and other virulence genes. The results of studies of invasion and survival of B. pertussis bacteria in eukaryotic cells, a change in B. pertussis bacteria population after experimental infection of laboratory mice and monkeys are presented, accumulation of avirulent insertion Bvg mutants of B. pertussis was detected. The data obtained are in accordance with the results of analysis of causative agent population in patients with typical and atypical forms of pertussis in humans. More than 50% of the population of B. pertussis bacteria in practically healthy carriers was shown to be presented by avirulent insertion Bvg mutants. B. pertussis virulence reducing as a result of inactivation of single or several virulence genes probably provide long-term persistence of bacteria in host organism and formation of apparently healthy vehicles. Follow-up studies on that front would help to formulate new attitudes to preventive measures of pertussis and lead to development of fundamentally new pharmaceuticals (vaccines) preventing formation of bacterial persistence. PMID:26951000

  6. Analusis by 252Cf plasma desorption mass spectrometry of Bordetella pertussis endotoxin after nitrous deamination

    Deprun, C.; Karibian, D.; Caroff, M.


    Endotoxic lipopolysaccharides (LPSs) are the major components of Gram-negative bacterial outer membrane. Like many amphipathic molecules, they pose problems of heterogeneity, purity, solubility, and aggregation. Nevertheless, PDMS has recently have been applied to unmodified endotoxins composed of LPS having uip to five sugar units in their saccharide chain. The B. Pertussis LPSs, most of which have a dodecasaccharide domain, ahve been analysed by classical methods and the masses of the separate lipid and saccharide domains determined after rupture of the bond linking them. However, the acid treatment employed for these and most chemical analyses can also modify structures in the vicinity of the bond. In order to investigate this biologically-important region, the endotoxin was treated to nitrous deamination, which shortens the saccharide chain to five sugars, but preserves the acid-labile region of the LPS. The PDM spectrum of this derivative, which required new conditions for its desorption, confirmed the structure analysis and demonstrated the presence of at least four molecular species.

  7. Oligomerization is involved in pore formation by Bordetella adenylate cyclase toxin

    Vojtová, Jana; Basler, Marek; Osička, Radim; Knapp, O.; Maier, E.; Černý, J.; Benada, Oldřich; Benz, R.; Šebo, Peter


    Roč. 23, - (2009), s. 2831-2843. ISSN 0892-6638 R&D Projects: GA AV ČR IAA500200914; GA MŠk 1M0506 Grant ostatní: -(XE) LSHB-CT-2003-503582 THERAVAC Institutional research plan: CEZ:AV0Z50200510 Keywords : blue native electrophoresis * planar lipid bilayer membranes * pore-forming activity Subject RIV: EE - Microbiology, Virology Impact factor: 6.401, year: 2009

  8. Bordetella pertussis epidemiology and evolution in the light of pertussis resurgence.

    Sealey, Katie L; Belcher, Thomas; Preston, Andrew


    Whooping cough, or pertussis, is resurgent in many countries world-wide. This is linked to switching from the use of whole cell vaccines to acellular vaccines in developed countries. Current evidence suggests that this has resulted in the earlier waning of vaccine-induced immunity, an increase in asymptomatic infection with concomitant increases in transmission and increased selection pressure for Bordetellapertussis variants that are better able to evade vaccine-mediated immunity than older isolates. This review discusses recent findings in B. pertussis epidemiology and evolution in the light of pertussis resurgence, and highlights the important role for genomics-based studies in monitoring B. pertussis adaptation. PMID:26932577

  9. The RNA Chaperone Hfq Is Required for Virulence of Bordetella pertussis

    Bíbová, Ilona; Škopová, Karolína; Mašín, Jiří; Černý, Ondřej; Hot, D.; Šebo, Peter; Večerek, Branislav


    Roč. 81, č. 11 (2013), s. 4081-4090. ISSN 0019-9567 R&D Projects: GA ČR(CZ) GAP302/11/1940; GA ČR GAP302/12/0460 Institutional support: RVO:61388971 Keywords : ADENYLATE-CYCLASE TOXIN * ISLET-ACTIVATING PROTEIN * ESCHERICHIA-COLI HFQ Subject RIV: EC - Immunology Impact factor: 4.156, year: 2013

  10. Quantification of potassium levels in cells treated with Bordetella adenylate cyclase toxin

    Wald, Tomáš; Petry-Podgorska, Inga; Fišer, Radovan; Matoušek, Tomáš; Dědina, Jiří; Osička, Radim; Šebo, Peter; Mašín, Jiří


    Roč. 450, APR 2014 (2014), s. 57-62. ISSN 0003-2697 R&D Projects: GA ČR(CZ) GAP302/11/0580; GA ČR GA13-14547S; GA ČR GAP302/12/0460 Institutional support: RVO:61388971 ; RVO:68081715 Keywords : Potassium * Adenylate cyclase toxin * RTX Subject RIV: CE - Biochemistry Impact factor: 2.219, year: 2014

  11. [Cryofractographic study of intercellular junctions in the populations of agar-cultivated Bordetella pertussis].

    Vysotskiĭ, V V; Vaisman, I Sh; Efimova, O G; Chemurzieva, N V


    The characteristic feature of replicas obtained from the freeze-fractures of B. pertussis unfixed cultures developing on casein charcoal agar for 1-7 days is the associative growth of highly polymorphic cells, ensured by the ramified system of intercellular connections (IC) formed by the derivatives of the outer layers of the cell wall. This proves that the associative location of bacterial cells, linked by numerous IC, in the preparation is not the artefact appearing in the process of their chemical fixation. In replicas obtained from the freeze-fractures of B. pertussis cultures, previously fixed with glutaraldehyde, osmic acid and uranyl acetate, oval cells with the cytoplasm having a relatively homogeneous structure and with the smoothed-out three-layer cell wall prevail. As a rule, IC are limited to the sites of direct contacts between individual cells. PMID:2866645

  12. Internation of Bordetella pertussis Adenylate Cyclase with CD11b/CD18

    El-Azami-El-Idrisi, M.; Bauche, C.; Loucká, Jiřina; Osička, Radim; Šebo, Peter; Ladant, D.; Leclerc, C.


    Roč. 278, č. 40 (2003), s. 38514-38521. ISSN 0021-9258 R&D Projects: GA AV ČR IPP1050128; GA ČR GA310/01/0934; GA AV ČR IAA5020907 Grant ostatní: GA by National Institutes of Health Grant(XX) 55000334; GA QLK2-CT-1999(XX) 00556 Institutional research plan: CEZ:AV0Z5020903 Keywords : cyaa * rtx * cd11b Subject RIV: EE - Microbiology, Virology Impact factor: 6.482, year: 2003

  13. Interaction of Bordetella adenylate cyclase toxin with complement receptor 3 involves multivalent glycan binding

    Hasan, Shakir; Osičková, Adriana; Bumba, Ladislav; Novák, Petr; Šebo, Peter; Osička, Radim


    Roč. 589, č. 3 (2015), s. 374-379. ISSN 0014-5793 R&D Projects: GA ČR(CZ) GAP302/11/0580; GA ČR(CZ) GA15-09157S; GA ČR(CZ) GA15-11851S Institutional support: RVO:61388971 Keywords : Adenylate cyclase toxin * CD11b/CD18 * Complement receptor type 3 Subject RIV: CE - Biochemistry Impact factor: 3.169, year: 2014

  14. Inflammasome Activation by Adenylate Cyclase Toxin Directs Th17 Responses and Protection against Bordetella pertussis

    Dunne, A.; Ross, P. J.; Pospíšilová, Eva; Mašín, Jiří; Meaney, A.; Sutton, C. E.; Iwakura, Y.; Tschopp, J.; Šebo, Peter; Mills, K. H. G.


    Roč. 187, č. 3 (2010), s. 1711-1719. ISSN 0022-1767 R&D Projects: GA ČR GA310/08/0447; GA AV ČR IAA500200914 Institutional research plan: CEZ:AV0Z50200510 Keywords : ADAPTIVE IMMUNE-RESPONSES * IL-17-PRODUCING T-CELLS * HOST-DEFENSE Subject RIV: EC - Immunology Impact factor: 5.745, year: 2010

  15. Bordetella pertussis: an underreported pathogen in pediatric respiratory infections, a prospective cohort study

    Brink, van den G.; Wishaupt, J.O.; Douma, J.C.; Hartwig, N.G.; Versteegh, F.G.A.


    Background: The incidence of pertussis has been increasing worldwide. In the Netherlands, the seroprevalence has risen higher than the reported cases, suggesting that laboratory tests for pertussis are considered infrequently and that even more pertussis cases are missed. The objective of our study

  16. Bordetella adenylate cyclase toxin is a unique ligand of the integrin complement receptor 3

    Osička, Radim; Osičková, Adriana; Hasan, Shakir; Bumba, Ladislav; Černý, Jiří; Šebo, Peter


    Roč. 4, DEC 9 (2015). ISSN 2050-084X R&D Projects: GA ČR(CZ) GAP302/11/0580; GA ČR(CZ) GA15-11851S; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388971 ; RVO:86652036 Keywords : E. coli * adenylate cyclase toxin * biochemistry Subject RIV: CE - Biochemistry Impact factor: 9.322, year: 2014

  17. Acylation of lysine 983 is sufficient for toxin activity of Bordetella pertussis adenylate cyclase

    Basar, T.; Havlíček, Vladimír; Bezoušková, Silvia; Hackett, M.; Šebo, Peter


    Roč. 276, č. 1 (2001), s. 348-354. ISSN 0021-9258 R&D Projects: GA ČR GA310/98/0432; GA ČR GV310/96/K102; GA AV ČR IAA5020907; GA MŠk ME 167; GA MŠk VS96149 Institutional research plan: CEZ:A53/98:Z5-020-9ii Subject RIV: EE - Microbiology, Virology Impact factor: 7.258, year: 2001

  18. Genómica funcional de Bordetella pertussis, implicancias sobre una enfermedad considerada reemergente

    Gaillard, María Emilia


    Objetivos generales de la tesis: Con el desarrollo de esta propuesta se espera contribuir al conocimiento que sirva de base para el diseño de una vacuna más efectiva contra pertussis, no sólo en términos generales, sino en lo que se refiere a su efectividad en Argentina, determinando la definición de la/s cepas / componentes a incluir en una nueva formulación. Objetivos específicos de la tesis: En este marco conceptual y tomando como hipótesis la divergencia de la población bacterian...

  19. Bordetella adenylate cyclase toxin induces a cascade of morphological changes of sheep erythrocytes and localizes into clusters in erythrocyte membranes

    Vojtová, Jana; Kofroňová, Olga; Šebo, Peter; Benada, Oldřich


    Roč. 69, - (2006), s. 119-129. ISSN 1059-910X R&D Projects: GA AV ČR IAA5020406 Institutional research plan: CEZ:AV0Z50200510 Keywords : cyaa * scanning electron microscopy * transmission electron microscopy Subject RIV: EE - Microbiology, Virology Impact factor: 1.680, year: 2006

  20. Efficient Ex Vivo Stimulation of Mycobacterium tuberculosis-Specific T Cells by Genetically Detoxified Bordetella pertussis Adenylate Cyclase Antigen Toxodids

    Wilkinson, K. A.; Šimšová, Marcela; Schölvinck, E.; Šebo, Peter; Leclerc, C.; Voredermeier, H. M.; Dickson, S. J.; Brown, J. R.; Davidson, R. N.; Pasvol, G.; Levin, M.; Wilkinson, R. J.


    Roč. 73, č. 5 (2005), s. 2991-2998. ISSN 0019-9567 R&D Projects: GA AV ČR IBS5020311; GA ČR GA310/01/0934 Institutional research plan: CEZ:AV0Z50200510 Keywords : mycobacterium tuberculosis * t cell * CyaA Subject RIV: EE - Microbiology, Virology Impact factor: 3.933, year: 2005

  1. Bordetella bronchiseptica como un riesgo importante de salud publica. Estudio clínico patológico en conejos

    Valladares-Carranza B.


    Full Text Available ResumenB. bronchiseptica es reconocida como un patógeno primario inicial del tracto respiratorio en animales domésticos, puede provocar tos de las perreras (perro, respiración ruidosa (en conejos y rinitis atrófica (en el cerdo.summary

  2. Bordetella bronchiseptica como un riesgo importante de salud publica. Estudio clínico patológico en conejos

    Valladares-Carranza B.; Ortega-Santana C.; Velazquez-Ordoñez V; Zamora-Espinosa J.L.; Peñuelas-Rivas C.G.; Castro-Maruri J.; Talavera-Rojas M.; Alonso-Fresan M.U.; Zaragoza-Bastida A.


    ResumenB. bronchiseptica es reconocida como un patógeno primario inicial del tracto respiratorio en animales domésticos, puede provocar tos de las perreras (perro), respiración ruidosa (en conejos) y rinitis atrófica (en el cerdo).summary

  3. A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease

    Ivanov, Yury V.; Shariat, Nikki; Karen B Register; Linz, Bodo; Rivera, Israel; Hu, Kai; Dudley, Edward G.; Harvill, Eric T.


    Background Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated genes (cas) are widely distributed among bacteria. These systems provide adaptive immunity against mobile genetic elements specified by the spacer sequences stored within the CRISPR. Methods The CRISPR-Cas system has been identified using Basic Local Alignment Search Tool (BLAST) against other sequenced and annotated genomes and confirmed via CRISPRfinder program. Using Polymerase Chain Reactio...

  4. Crystallization and preliminary X-ray diffraction analysis of the peptidylprolyl isomerase Par27 of Bordetella pertussis

    Par27 from B. pertussis, the prototype of a new group of parvulins has been crystallized in two different crystal forms. Proteins with both peptidylprolyl isomerase (PPIase) and chaperone activities play a crucial role in protein folding in the periplasm of Gram-negative bacteria. Few such proteins have been structurally characterized and to date only the crystal structure of SurA from Escherichia coli has been reported. Par27, the prototype of a new group of parvulins, has recently been identified. Par27 exhibits both chaperone and PPIase activities in vitro and is the first identified parvulin protein that forms dimers in solution. Par27 has been expressed in E. coli. The protein was purified using affinity and gel-filtration chromatographic techniques and crystallized in two different crystal forms. Form A, which belongs to space group P2 (unit-cell parameters a = 42.2, b = 142.8, c = 56.0 Å, β = 95.1°), diffracts to 2.8 Å resolution, while form B, which belongs to space group C222 (unit-cell parameters a = 54.6, b = 214.1, c = 57.8 Å), diffracts to 2.2 Å resolution. Preliminary diffraction data analysis agreed with the presence of one monomer in the asymmetric unit of the orthorhombic crystal form and two in the monoclinic form

  5. Transcriptional profiling of Bordetella pertussis reveals requirement of RNA chaperone Hfq for Type III secretion system functionality

    Bíbová, Ilona; Hot, D.; Keidel, Kristina; Amman, F.; Slupek, S.; Černý, Ondřej; Gross, R.; Večerek, Branislav


    Roč. 12, č. 2 (2015), s. 175-185. ISSN 1547-6286 R&D Projects: GA ČR(CZ) GAP302/11/1940; GA MŠk(CZ) EE2.3.20.0055; GA MŠk(CZ) EE2.3.30.0003; GA MŠk(CZ) 7AMB14AR028; GA MŠk(CZ) ED1.1.00/02.0109 Institutional support: RVO:61388971 Keywords : Bsp22 * Hfq * infection Subject RIV: CE - Biochemistry Impact factor: 4.974, year: 2014

  6. Mass spectrometric analysis of recombinant adenylate cyclase toxin from Bordetella pertussis strain 18323/pHSP9+

    Havlíček, Vladimír; Higgins, L.; Chen, W.; Halada, Petr; Šebo, Peter; Sakamoto, H.; Hackett, M.


    Roč. 36, - (2001), s. 384-391. ISSN 1076-5174 R&D Projects: GA AV ČR IAA5020907; GA MŠk ME 167; GA MŠk VS96141 Institutional research plan: CEZ:A53/98:Z5-020-9ii Subject RIV: EE - Microbiology, Virology Impact factor: 2.685, year: 2001

  7. A newly discovered Bordetella species carries a transcriptionally active CRISPR-Cas with a small Cas9 endonuclease

    The Cas9 endonuclease of the Type II-a clustered regularly interspersed short palindromic repeats (CRISPR), of Streptococcus pyogenes (SpCas9) has been adapted as a widely used tool for genome editing and genome engineering. Herein, we describe a gene encoding a novel Cas9 ortholog (BpsuCas9) and th...

  8. The adenylate cyclase toxin from Bordetella pertussis - a novel promising vehicke fer antigen delivery to dendritic cells

    Šimšová, Marcela; Šebo, Peter; Leclerc, C.


    Roč. 293, - (2004), s. 571-576. ISSN 1438-4221 R&D Projects: GA ČR GA310/01/0934; GA AV ČR IAA5020907 Grant ostatní: GA QLK2-CT-1999(XX) 00556 Keywords : cyaa * cellular immune response * antigen delivery Subject RIV: EE - Microbiology, Virology Impact factor: 2.919, year: 2004

  9. Bulk protein biosynthesis of the spleen and some splenic cell populations after induction of splenomegaly by application of Bordetella pertussis

    Autoradiographic studies and liquid scintillation counting were carried out in female NMRI mice just reaching maturity. All animals had received a single injection, either of bovine serum albumin (BSA) or of pertussis organism (PO) or BSA + PO. The animals were sacrificed 4 d and 10 d after this pretreatment. 2 h before decapitation, a single dose of 3H-l phenyl alamine was applied intraperitoneally. The following results were obtained: The splenic index (splenic weight in mg/mouse weight in g) increased as a result of splenomegaly caused by PO. Morphometric data suggested an enlarged cell and nuclear area with enhanced cellular amino acid turnover and migration of RNP-containing matter into the nucleus, especially in the megakaryocytes and in lymphocytoid blastic cells. Incorporation of 3H-l-phenylalanine per unit of dry weight of the spleen is slowed down during the experiment while amiro acid incorporation by the total spleen increases with PO-induced splenomegaly. Incorporation of amino acid per unit of dry weight is constant in all experimental and control animals. The increased amino acid incorporation in lymphocytoid blastic cells is probably caused by the immunological situations during the experiment. An explanation of total cell increase and cell increase of megakaryocytic splenic cells is attempted. (orig./MG)

  10. A CpG-containing oligodeoxynucleotide adjuvant for acellular pertussis vaccine improves the protective response against Bordetella pertussis

    Asokanathan, Catpagavalli; Corbel, Michael; Xing, Dorothy


    We investigated the adjuvant effect of CpG ODN alone or in combination with aluminum hydroxide on the immune response to the three main antigens presented in current acellular pertussis vaccines: pertussis toxoid, filamentous haemagglutinin and pertactin. The development of protection in mice was investigated for the intra-peritoneal and intra-nasal immunisation routes. The results showed that CpG ODN alone, or in combination with aluminum hydroxide, gave enhancement in anti-pertussis toxin, ...

  11. Disease: H01077 [KEGG MEDICUS

    Full Text Available ts or patients with underlying disease. Infectious disease Bordetella hinzii Amoxic... the genus Bordetella that is isolated from poultry with respiratory disease. B. hinzii may cause disease in immunocompromised patien

  12. Disease: H01083 [KEGG MEDICUS

    Full Text Available 73090 (description, env_factor) Meis JF, van Griethuijsen AJ, Muytjens HL Bordetella bronchiseptica bronchitis...env_factor) Papasian CJ, Downs NJ, Talley RL, Romberger DJ, Hodges GR Bordetella bronchiseptica bronchitis

  13. Disease: H01066 [KEGG MEDICUS

    Full Text Available rriott D Bordetella petrii from a clinical sample in Australia: isolation and mol...lar pathogenesis, epidemiology, and clinical manifestations of respiratory infections due to Bordetella pertussis and other Bordetella subspecies. Clin Microbiol Rev 18:326-82 (2005) ...

  14. Bordetella Adenylate Cyclase Toxin Differentially Modulates Toll-Like Receptor-Stimulated Activation, Migration and T Cell Stimulatory Capacity of Dendritic Cells

    Adkins, Irena; Kamanová, Jana; Kocourková, A.; Švédová, Martina; Tomala, Jakub; Janová, H.; Mašín, Jiří; Chládková, Barbara; Bumba, Ladislav; Kovář, Marek; Ross, P. J.; Tučková, Ludmila; Spíšek, R.; Mills, K. H. G.; Šebo, Peter


    Roč. 9, č. 8 (2014). E-ISSN 1932-6203 R&D Projects: GA ČR GA310/08/0447; GA ČR GP310/09/P582; GA ČR GAP301/11/0325; GA MŠk 1M0506 Institutional support: RVO:61388971 Keywords : RESPIRATORY-INFECTION * INTERLEUKIN-10 PRODUCTION * PROTECTIVE IMMUNITY Subject RIV: EE - Microbiology, Virology Impact factor: 3.234, year: 2014

  15. Bordetella pertussis commits human dendritic cells to promote a Th1/Th17 response through the activity of adenylate cyclase toxin and MAPK-pathways.

    Giorgio Fedele

    Full Text Available The complex pathology of B. pertussis infection is due to multiple virulence factors having disparate effects on different cell types. We focused our investigation on the ability of B. pertussis to modulate host immunity, in particular on the role played by adenylate cyclase toxin (CyaA, an important virulence factor of B. pertussis. As a tool, we used human monocyte derived dendritic cells (MDDC, an ex vivo model useful for the evaluation of the regulatory potential of DC on T cell immune responses. The work compared MDDC functions after encounter with wild-type B. pertussis (BpWT or a mutant lacking CyaA (BpCyaA-, or the BpCyaA- strain supplemented with either the fully functional CyaA or a derivative, CyaA*, lacking adenylate cyclase activity. As a first step, MDDC maturation, cytokine production, and modulation of T helper cell polarization were evaluated. As a second step, engagement of Toll-like receptors (TLR 2 and TLR4 by B. pertussis and the signaling events connected to this were analyzed. These approaches allowed us to demonstrate that CyaA expressed by B. pertussis strongly interferes with DC functions, by reducing the expression of phenotypic markers and immunomodulatory cytokines, and blocking IL-12p70 production. B. pertussis-treated MDDC promoted a mixed Th1/Th17 polarization, and the activity of CyaA altered the Th1/Th17 balance, enhancing Th17 and limiting Th1 expansion. We also demonstrated that Th1 effectors are induced by B. pertussis-MDDC in the absence of IL-12p70 through an ERK1/2 dependent mechanism, and that p38 MAPK is essential for MDDC-driven Th17 expansion. The data suggest that CyaA mediates an escape strategy for the bacterium, since it reduces Th1 immunity and increases Th17 responses thought to be responsible, when the response is exacerbated, for enhanced lung inflammation and injury.

  16. ε-Caprolactam Utilization by Proteus sp. and Bordetella sp. Isolated From Solid Waste Dumpsites in Lagos State, Nigeria, First Report

    Sanuth, Hassan Adeyemi; Yadav, Amit; FAGADE, Obasola Ezekiel; Shouche, Yogesh


    The ε-caprolactam is the monomer of the synthetic non-degradable nylon-6 and often found as nonreactive component of nylon-6 manufacturing waste effluent. Environmental consequences of its toxicity to natural habitats and humans pose a global public concern. Soil samples were collected from three designated solid waste dumpsites, namely, Abule-Egba, Olusosun and Isheri-Igando in Lagos State, Nigeria. Sixteen bacteria isolated from these samples were found to utilize the ε-caprolactam as a sol...

  17. Synthesis of acyclic nucleoside phosphonates bearing (N-methyl)anthraniloyl substituent as potential inhibitors of adenylate cyclase toxin from Bordetella Pertussis

    Břehová, Petra; Šmídková, Markéta; Mertlíková-Kaiserová, Helena; Dračínský, Martin; Janeba, Zlatko

    Praha: Czech Chemical Society, 2015. s. 61. [Liblice 2015. Advances in Organic , Bioorganic and Pharmaceutical Chemistry /50./. 06.11.2015-08.11.2015, Olomouc] R&D Projects: GA MV VG20102015046 Institutional support: RVO:61388963 Keywords : acyclic nucleoside phosphonates * adenylate cyclase toxin * prodrugs Subject RIV: CC - Organic Chemistry

  18. Complete protection against P. berghei malaria upon heterologous prime/boost immunization against circumsporozoite protein employing Salmonella type III secretion system and Bordetella adenylate cyclase toxoid

    Tartz, S.; Rüssmann, H.; Kamanová, Jana; Šebo, Peter; Sturm, A.; Heussler, V.; Fleischer, B.; Jacobs, T.


    Roč. 26, č. 47 (2008), s. 5935-5943. ISSN 0264-410X R&D Projects: GA MŠk 2B06161 Institutional research plan: CEZ:AV0Z50200510 Keywords : circumsporozoite protein * vaccine * salmonella Subject RIV: EE - Microbiology, Virology Impact factor: 3.298, year: 2008

  19. Bordetella pertussis Adenylate Cyclase Toxin Blocks Induction of Bactericidal Nitric Oxide in Macrophages through cAMP-Dependent Activation of the SHP-1 Phosphatase

    Černý, Ondřej; Kamanová, Jana; Mašín, Jiří; Bíbová, Ilona; Škopová, Karolína; Šebo, Peter


    Roč. 194, č. 10 (2015), s. 4901-4913. ISSN 0022-1767 R&D Projects: GA ČR GAP302/12/0460; GA ČR GA13-14547S Institutional support: RVO:61388971 Keywords : CYCLIC-AMP * MURINE MACROPHAGES * IFN-GAMMA Subject RIV: EE - Microbiology, Virology Impact factor: 4.922, year: 2014

  20. Bordetella Adenylate Cyclase Toxin Mobilizes Its beta(2) Integrin Receptor into Lipid Rafts to Accomplish Translocation across Target Cell Membrane in Two Steps

    Bumba, Ladislav; Mašín, Jiří; Fišer, R.; Šebo, Peter


    Roč. 6, č. 5 (2010), s. 1-15. ISSN 1553-7366 R&D Projects: GA ČR GP310/07/P115; GA MŠk 1M0506; GA AV ČR IAA500200914; GA MŠk 2B06161; GA ČR GA310/08/0447 Institutional research plan: CEZ:AV0Z50200510; CEZ:AV0Z50520701 Keywords : SHEEP ERYTHROCYTES * BACILLUS -ANTHRACIS * ESCHERICHIA-COLI Subject RIV: EE - Microbiology, Virology Impact factor: 9.079, year: 2010

  1. Prime/boost immunotherapy of HPV16-induced tumors with E7 protein delivered by Bordetella adenylate cyclase and modified vaccinia virus Ankara

    Macková, J.; Stasíková, J.; Kutinová, L.; Mašín, Jiří; Hainz, P.; Šimšová, Marcela; Gabriel, P.; Šebo, Peter; Němečková, P.


    Roč. 55, - (2006), s. 39-46. ISSN 0340-7004 R&D Projects: GA AV ČR IBS5020311; GA ČR GA310/04/0004; GA MZd NR8004 Grant ostatní: GA MZd NC6570 Institutional research plan: CEZ:AV0Z50200510 Keywords : vaccine * hpv-e7 * vaccinia virus Subject RIV: EE - Microbiology, Virology Impact factor: 4.313, year: 2006

  2. Bordetella adenylate cyclase toxin: a unique combination of a pore-forming moiety with a cell-invading adenylate cyclase enzyme

    Mašín, Jiří; Osička, Radim; Bumba, Ladislav; Šebo, Peter


    Roč. 73, č. 8 (2015). ISSN 2049-632X R&D Projects: GA ČR GAP302/12/0460; GA ČR GA15-09157S; GA ČR(CZ) GA15-11851S Institutional support: RVO:61388971 Keywords : adenylate cyclase toxin * membrane penetration * pore-formation Subject RIV: EE - Microbiology, Virology Impact factor: 2.403, year: 2014

  3. Delivery of Large Heterologous Polypeptides across the Cytoplasmic Membrane of Antigen-Presenting Cells by the Bordetella RTX Hemolysin Moiety Lacking the Adenylyl Cyclase Domain

    Holubová, Jana; Kamanová, Jana; Jelínek, J.; Tomala, Jakub; Mašín, Jiří; Kosová, Martina; Staněk, Ondřej; Bumba, Ladislav; Michálek, J.; Kovář, Marek; Šebo, Peter


    Roč. 80, č. 3 (2012), s. 1181-1192. ISSN 0019-9567 R&D Projects: GA AV ČR IAA500200914; GA ČR(CZ) GAP207/11/0717; GA ČR GAP301/11/0325; GA MŠk 1M0506; GA MŠk 2B06161 Institutional research plan: CEZ:AV0Z50200510 Keywords : MHC CLASS-I * ESCHERICHIA-COLI * PRESENTATION PATHWAY Subject RIV: EE - Microbiology, Virology Impact factor: 4.074, year: 2012

  4. Cross-reactions in IgM ELISA tests to Legionella pneumophila sg1 and Bordetella pertussis among children suspected of legionellosis; potential impact of vaccination against pertussis?


    The objective of this study was preliminary evaluation of IgM cross-reaction in sera collected from children hospitalized because of suspected legionellosis. Sera with positive IgM results to L. pneumophila sgs1-7, B. pertussis or with simultaneous detection of IgM antibodies to L. pneumophila sgs1-7 and B. pertussis, or IgM to L. pneumophila sgs1-7 and M. pneumoniae in routine tests, were selected. In total, an adapted pre-absorption test was used for the serological confirmation of legionellosis in the sera of 19 children suspected of legionellosis, and also in 3 adult persons with confirmed Legionnaires’ disease. Sera were pre-absorbed with antigens of L. pneumophila sg1, B. pertussis or both, and tested by ELISA tests. The reduction of IgM antibody level by pre-absorption with antigen/antigens was determined. Reduction of anti-Lpsgs1-7 IgM by pre-absorption with L.pneumophila sg1 antigen ranged from 1.5 to 80, and reduction of anti-Bp IgM by pre-absorption with B. pertussis ranged from 2.0 to 23.8. Reduction by both antigens varied depending on the age of the patients: among children <4 yrs.old, the reduction of anti-B. pertussis IgM by both antigens was higher than for B. pertussis antigen alone. Based on the high difference (≥ 2 times) between reduction by L.pneumophila sg1 and by B. pertussis antigen, legionellosis was confirmed in 8/19 children. The majority of them also indicated IgM positive/borderline results for B. pertussis or M.pneumoniae in routine ELISA tests. As a preliminary, we posed a hypothesis of a potential impact of an anti-pertussis vaccination on the results obtained in anti-L. pneumophila ELISA IgM tests among young children. PMID:26557032

  5. 78 FR 19986 - New Animal Drugs; Enrofloxacin; Tilmicosin; Tylosin


    ..., Pasteurella multocida, Haemophilus parasuis, Streptococcus suis, Bordetella bronchiseptica, and Mycoplasma hyopneumoniae. (B) For the treatment and control of swine respiratory disease (SRD) associated...

  6. NCBI nr-aa BLAST: CBRC-STRI-01-2656 [SEVENS

    Full Text Available CBRC-STRI-01-2656 ref|NP_880118.1| negative regulator of flagellin synthesis ... [Bordetella pertuss ... I] emb|CAE41662.1| negative regulator of flagellin synthesis ... [Bordetella pertussis Tohama I] NP_880118.1 9.2 40 ...

  7. 77 FR 76862 - New Animal Drugs; Enrofloxacin; Melengestrol; Meloxicam; Pradofloxacin; Tylosin


    ... parasuis, Streptococcus suis, Bordetella bronchiseptica, and Mycoplasma hyopneumoniae. * * * * * PART 529.... respiratory disease Shawnee Mission, associated with KS 66201. Bordetella bronchiseptica and Mycoplasma hyopneumoniae. 200-534 Huvepharma AD, 5th TYLOVET 100 Original approval as a 558.342 Yes CE \\1\\ Floor,...

  8. Immunization with a Circumsporozoite Epitope Fused to Bordetella pertussis Adenylate Cyclase in Conjunction with Cytotoxic T-Lymphocyte-Associated Antigen 4 Blockade Confers Protection against Plasmodium berghei Liver-Stage Malaria

    Tartz, S.; Kamanová, Jana; Šimšová, Marcela; Šebo, Peter; Bolte, S.; Heussler, V.; Fleischer, B.; Jacobs, T.


    Roč. 74, č. 4 (2006), s. 2277-2285. ISSN 0019-9567 R&D Projects: GA AV ČR IBS5020311 Institutional research plan: CEZ:AV0Z50200510 Keywords : plasmodium berghei * immunity * malaria Subject RIV: EE - Microbiology, Virology Impact factor: 4.004, year: 2006

  9. The Bordetella pertussis Type III Secretion System Tip Complex Protein Bsp22 Is Not a Protective Antigen and Fails To Elicit Serum Antibody Responses during Infection of Humans and Mice

    Villarino Romero, Rodrigo; Bíbová, Ilona; Černý, Ondřej; Večerek, Branislav; Wald, Tomáš; Benada, Oldřich; Zavadilová, J.; Osička, Radim; Šebo, Peter


    Roč. 81, č. 8 (2013), s. 2761-2767. ISSN 0019-9567 R&D Projects: GA ČR(CZ) GAP302/11/0580; GA ČR(CZ) GAP302/11/1940 Institutional support: RVO:61388971 Keywords : ADENYLATE CYCLASE-HEMOLYSIN * T-CELL EPITOPES * IMMUNE-RESPONSES Subject RIV: EC - Immunology Impact factor: 4.156, year: 2013

  10. Acylation of conserved lysine 983 is sufficient for activity of Bordetella adenylate cyclase toxin: substitutions of alanine 140 in the acyltransferase CyaC modulate the selection of toxin acylation sites

    Basar, T.; Havlíček, Vladimír; Bezoušková, Silvia; Higgins, L.; Hackett, M.; Šebo, Peter


    Roč. 94, č. 8 (2000), s. 568. ISSN 0009-2770. [Biochemický sjezd /17./. 07.09.2000-10.09.2000, Prague] R&D Projects: GA ČR GA310/98/0432; GA AV ČR IAA5020907; GA MŠk VS96149; GA MŠk ME 167 Subject RIV: CE - Biochemistry

  11. Dicty_cDB: Contig-U08276-1 [Dicty_cDB

    Full Text Available s vulgatus ATCC 8482,... 68 5e-10 CP000937_1762( CP000937 |pid:none) Francisella philomiragia subsp.... 68 5...ncisella philomiragia subsp.... 68 7e-10 CP000387_341( CP000387 |pid...:none) Bordetella bronchiseptica strain... 66 3e-09 AY184800_1( AY184800 |pid:none) Chlam...:none) Streptococcus uberis 0140J comp... 65 3e-09 AM902716_1980( AM902716 |pid:none) Bordetella petrii stra...5 6e-09 AE015928_2229( AE015928 |pid:none) Bacteroides thetaiotaomicron VP... 65 6e-09 BX640414_260( BX640414 |pid:none) Bordetella

  12. 21 CFR 558.630 - Tylosin and sulfamethazine.


    ... bronchiseptica rhinitis; prevention of swine dysentery associated with Brachyspira hyodysenteriae; and control of... presence of atrophic rhinitis; lowering the incidence and severity of Bordetella bronchiseptica rhinitis; prevention of swine dysentery (vibrionic); control of swine pneumonias caused by bacterial...

  13. Detection of respiratory pathogens in aerosols from acutely infected pigs

    Infectious agents that cause respiratory disease in pigs include porcine reproductive respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), swine influenza virus (SIV), porcine respiratory corona virus (PRCV), Mycoplasma hyopneumoniae, and Bordetella bronchiseptica. The objective of...

  14. GenBank blastx search result: AK288959 [KOME

    Full Text Available AK288959 J090084E19 DQ402420.1 DQ402420 Bordetella holmesii molecular ... chaperone (dnaJ) gene, par ... tial cds; molecular ... chaperone (dnaK) and putative thioredoxin genes, c ...

  15. GenBank blastx search result: AK288072 [KOME

    Full Text Available AK288072 J075161I05 DQ402420.1 DQ402420 Bordetella holmesii molecular ... chaperone (dnaJ) gene, par ... tial cds; molecular ... chaperone (dnaK) and putative thioredoxin genes, c ...

  16. AcEST: DK946704 [AcEST

    Full Text Available 197 >sp|Q7WHN1|SYH_BORBR Histidyl-tRNA synthetase OS=Bordetella bronchiseptica GN=his...YMU02A01NGRL0013_H09 206 Adiantum capillus-veneris mRNA. clone: YMU02A01NGRL0013_H09. 5' end seq...on Adiantum capillus-veneris mRNA. clone: YMU02A01NGRL0013_H09. 5' end sequence. Accession DK946704 Tissue Q7W6P7 Definition sp|Q7W6P7|SYH_BORPA Histidyl-tRNA synthetase OS=Bordetella parapertussis Align length 3...nt alignments: (bits) Value sp|Q7W6P7|SYH_BORPA Histidyl-tRNA synthetase OS=Bordetella parap... 29 8.3 sp|Q7WHN1|SYH_BORBR His

  17. Dicty_cDB: Contig-U15255-1 [Dicty_cDB

    Full Text Available :none) Ralstonia eutropha JMP134 chrom... 97 5e-19 AP006841_3502( AP006841 |pid:none) Bacteroides 90 6e-17 CP000447_2048( CP000447 |pid:none) Shewanella frigidimarina NCIMB ... 90 6e-17 CS360325_1( CS360325 |pid... 7e-14 BX640416_135( BX640416 |pid:none) Bordetella pertussis strain Toha... 79 2e-13 CP001614_1734( CP001614 |pid...:none) Teredinibacter turnerae T7901, ... 78 3e-13 AM902716_379( AM902716 |pid:none) Bordetella petrii DSM 128... 77 4e-13 BX640429_59( BX640429 |pid:none) Bordetella parapertussis stra

  18. Characterization of two Achromobacter xylosoxidans isolates from patients with pertussis-like symptoms

    Fiorella; Orellana-Peralta; Michelle; Jacinto; Maria; J.Pons; Cláudia; Gomes; Carlos; Bada; Isabel; Reyes; Juana; del; Valle; Mendoza; Joaquim; Ruiz


    Objective:To characterize two Achromobaeter xylosoxidans recovered from 2 patients diagnosed with pertussis during a Bordetella pertussis surveillance program.Methods:Nasopharyngeal swabs from 2 children under 1 year of age with clinical suspicion of pertussis were analyzed by culture and PCR.Results:Two Achromobaeter xylosoxidans A8,closely related to Bordetella spp.were recovered from 2 patients diagnosed of pertussis,both carrying the ptxA gene and IS418 the pertussis toxin encoding gene.Subsequently,antibiotic susceptibility was evaluated by disk-diffusion method and by PCR.Conclusions:Although more detailed studies are needed,the present data highlight the possibility that Achromobaeter xylosoxidans.closely related Bordetella pertussis microorganisms and not covered under the vaccine umbrella,might also result in cases of whooping cough.Thereby further surveillance is necessary to determine the extension and relevance of their pathogenic role in order to discriminate their real public health implication.

  19. Characterization of two Achromobacter xylosoxidans isolates from patients with pertussis-like symptoms

    Fiorella Orellana-Peralta; Michelle Jacinto; Maria J Pons; Cludia Gomes; Carlos Bada; Isabel Reyes; Juana del Valle Mendoza; Joaquim Ruiz


    Objective:To characterize two Achromobacter xylosoxidans recovered from 2 patients diagnosed with pertussis during a Bordetella pertussis surveillance program. Methods:Nasopharyngeal swabs from 2 children under 1 year of age with clinical suspicion of pertussis were analyzed by culture and PCR. Results:Two Achromobacter xylosoxidans A8, closely related to Bordetella spp. were recovered from 2 patients diagnosed of pertussis, both carrying the ptxA gene and IS418 the pertussis toxin encoding gene. Subsequently, antibiotic susceptibility was evaluated by disk-diffusion method and by PCR. Conclusions:Although more detailed studies are needed, the present data highlight the possibility that Achromobacter xylosoxidans, closely related Bordetella pertussis microorganisms and not covered under the vaccine umbrella, might also result in cases of whooping cough. Thereby further surveillance is necessary to determine the extension and relevance of their pathogenic role in order to discriminate their real public health implication.

  20. Dicty_cDB: Contig-U11233-1 [Dicty_cDB

    Full Text Available 017332 |pid:none) Mycoplasma hyopneumoniae 232, com... 88 1e-15 BX640430_239( BX640430 |pid:none) Bordetella parapertussis strain... ... 88 1e-15 BX640421_259( BX640421 |pid:none) Bordetella pertussis strain Toha... 88 ...C=; ... 60 4e-07 AY596297_1840( AY596297 |pid:none) Haloarcula marismortui...sfrfidkvnre*rlqilnhhk*rfsrfigfrcr*erfil*ypmsy ydnplhvts*iflirfiti*rf*slwfgh*ylhsfhitnsslsryysssfisfcnwysis fikfre*nhfsinre**fps*ngsir...**syc*nl c****ss*skikn*lykskyfirti**y****fr*yfksrk*y**inflylifk*ifks* ktt*kyk

  1. Optimizing polymerase chain reaction testing for the diagnosis of pertussis: current perspectives

    Arbefeville S


    Full Text Available Sophie Arbefeville, Patricia Ferrieri Department of Laboratory Medicine and Pathology, University of Minnesota Medical School, Minneapolis, MN, USA Abstract: Nucleic acid testing has revolutionized the diagnosis of pertussis in the clinical microbiology laboratory and has become the main avenue of testing for pertussis infection. Real-time polymerase chain reaction (RT-PCR is an important tool for timely diagnosis of pertussis and is more sensitive than culture. The most commonly amplified targets are the insertion-sequence (IS genes, which are found in multiple copies in the genome of Bordetella species. Some strains of Bordetella pertussis have more than 200 copies of IS481 in their genome. This high number of repeats allows RT-PCR assays to be very sensitive and makes nucleic acid testing two to three times more sensitive than culture. Despite these advantages, RT-PCR can give inaccurate results due to contamination or lack of specificity. Contamination can easily happen during specimen collection, DNA extraction, or nucleic acid amplification steps. To avoid contamination, laboratories need to have quality controls and good workflows in place. The poor specificity of the nucleic acid assays amplifying the IS genes is because they are found in various Bordetella species and, thus, not unique to a specific species. Bordetella holmesii, a more recently described Bordetella species found to be responsible for respiratory symptoms similar to pertussis in adolescents and adults, can be misidentified as B. pertussis in RT-PCR assays that amplify only the IS481 target. Use of multiple targets may improve specificity of RT-PCR assays for pertussis. In the past few years, the US Food and Drug Administration has cleared three commercial assays for the detection of B. pertussis in respiratory specimens. Several commercial assays and analyte-specific reagents, which are not US Food and Drug Administration cleared, are available for the detection of one

  2. Dicty_cDB: Contig-U10024-1 [Dicty_cDB

    Full Text Available ndocina ymp, compl... 67 1e-09 CP000937_1274( CP000937 |pid:none) Francisella philomiragia subsp.... 67 1e-0.... 67 2e-09 AY445932_1( AY445932 |pid:none) Ctenopharyngodon idella disulfide ... ... CK721933 ) tad67f10.y2 Hydra EST -Kiel 1 Hydra magnipapillat... 72 5e-08 1 ( CJ416909 ) Molgula...(P6692... 67 2e-09 BX640446_284( BX640446 |pid:none) Bordetella bronchiseptica strain... 67 2e-09 AE015451_504( AE015451 |pid...6 3e-09 AM902716_1980( AM902716 |pid:none) Bordetella petrii strain DSM 12... 66

  3. Dicty_cDB: Contig-U15792-1 [Dicty_cDB

    Full Text Available baltica OS155, compl... 249 3e-64 AE014299_1859( AE014299 |pid:none) Shewanella oneidensis MR-1, com... 2... loihica PV-4, comple... 242 3e-62 CP000447_2708( CP000447 |pid:none) Shewanella frigidima... bronchiseptica strain ... 213 2e-53 BX640436_178( BX640436 |pid:none) Bordetella...s fragilis NCTC 9343,... 192 4e-47 AP006841_1611( AP006841 |pid:none) Bacteroides fragi...rensis subsp. novicida U112, comp... 52 0.069 1 ( CP000437 ) Francisella tularensis subsp. hola

  4. Disease: H01084 [KEGG MEDICUS

    Full Text Available e cause of bacteremia, pertussis-like respiratory tract infection, and endocarditis predominantly in patients... FR, Bruisten S, Linde I, Reubsaet F, Heuvelman K, van der Lee S, J King A Characterization of Bordetella holmesii isolates from pati...ents with pertussis-like illness in the Netherlands. FEM

  5. Pertussis specific T-cell immunity in Dutch children: Differences after whole-cell versus acellular vaccination

    Schure, R.M.


    Bordetella pertussis is the causative bacteria of whooping cough. Whooping cough is a highly contagious infection, which is characterized by coughing with whooping and post-tussive vomiting. In particular, infants under 6 months of age who have not been fully vaccinated, are at risk for serious comp

  6. Läkaköha - aktuaalne uurimisteema / Marje Oona

    Oona, Marje, 1963-


    TÜ peremeditsiini õppetooli töötajate poolt algatatud uurimistööst, mille eesmärgiks on uurida Bordetella spp. infektsioonide epidemioloogiat, molekulaargeneetikat ja kliinilisi eripärasid ning selgitada läkaköha sagedasema diagnsimise põhjusi Eestis

  7. Detection of respiratory pathogens in air samples from acutely infected pigs

    Hermann, Joseph R.; Brockmeier, Susan L.; Yoon, Kyoung-Jin; Zimmerman, Jeffrey J.


    Pathogens causing significant respiratory disease in growing pigs include Porcine reproductive and respiratory syndrome virus, Porcine circovirus 2, swine influenza virus, porcine respiratory coronavirus, Mycoplasma hyopneumoniae, and Bordetella bronchiseptica. The objective of this research was to characterize the respiratory excretion of these pathogens by acutely infected pigs. Pigs were inoculated under experimental conditions with 1 pathogen. Samples were collected from the upper respira...

  8. 21 CFR 522.2630 - Tulathromycin.


    .... multocida, Bordetella bronchiseptica, Haemophilus parasuis, and Mycoplasma hyopneumoniae; and for the control of SRD associated with A. pleuropneumoniae, P. multocida, and M. hyopneumoniae in groups of pigs..., Histophilus somni, and Mycoplasma bovis. For the control of respiratory disease in cattle at high risk...

  9. Immunological Links to Nonspecific Effects of DTwP and BCG Vaccines on Infant Mortality

    Claesson, Mogens Helweg


    A number of mainly observational studies suggest that many African females below the age of one year die each year from the nonspecific effects of vaccination with diphtheria-tetanus toxoids and killed (whole-cell) Bordetella pertussis (DTwP). In contrast, similar studies suggest that many African...

  10. Adults with pertussis

    MacLean, D. W.


    Eighty adults were diagnosed in one general practice as having infection due to Bordetella pertussis, type 1.3, during a period of 30 months. Their clinical presentation and progress is recorded. A plea is made for attention to be paid to this infection in adults.

  11. Whooping cough in South-East Romania: a 1-year study.

    Dinu, Sorin; Guillot, Sophie; Dragomirescu, Cristiana Cerasella; Brun, Delphine; Lazăr, Stefan; Vancea, Geta; Ionescu, Biatrice Mariana; Gherman, Mariana Felicia; Bjerkestrand, Andreea-Florina-Dana; Ungureanu, Vasilica; Guiso, Nicole; Damian, Maria


    The incidence of whooping cough in Romania is substantially underestimated, and, as noted by the health authorities, this is mostly due to the lack of both awareness and biological diagnosis. We conducted a 1-year study in Bucharest in order to assess the circulation of Bordetella pertussis, the main etiological agent of whooping cough. Fifty-one subjects suspected of whooping cough were enrolled. Culture, real-time PCR, and enzyme-linked immunosorbent assay were used for laboratory diagnosis. Whooping cough patients (63%) were distributed among all age groups, and most were unvaccinated, incompletely vaccinated, or had been vaccinated more than 5 years previously. Bordetella holmesii DNA was detected in 22% of the bordetellosis cases; these patients included adults; teenagers; and, surprisingly, young children. B. pertussis isolates were similar to the clinical isolates currently circulating elsewhere in Europe. One isolate does not express pertactin, an antigen included in some acellular pertussis vaccines. PMID:24355701

  12. Dicty_cDB: Contig-U06085-1 [Dicty_cDB

    Full Text Available 009 CP000937_1795( CP000937 |pid:none) Francisella philomiragia subsp.... 44 0.01...-10 AC116305_14( AC116305 |pid:none) Dictyostelium discoideum chromoso... 67 2e-10 CR628336_2207( CR628336 |pid:none) Legionella... tdn operon, pa... 40 0.12 BX640415_112( BX640415 |pid:none) Bordetella pertussis strain Toha... 40 0.15 AL161577_17( AL161577 |pid...te AToL-Lep-ID... 40 0.15 BX640445_2( BX640445 |pid:none) Bordetella...=GMP synthase [glutamine-hydrolyzing]; ... 34 8.4 CP000238_500( CP000238 |pid:none) Baumannia cicadellinicola

  13. Dicty_cDB: Contig-U14933-1 [Dicty_cDB

    Full Text Available :none) Saccharophagus degradans 2-40, ... 140 2e-55 CP000444_1874( CP000444 |pid:none) Shewanella...e-55 AP006841_4096( AP006841 |pid:none) Bacteroides fragilis YCH46 DNA,... 148 3e-55 AP008955_517( AP008955 |pid:none) Brevibacill...:none) Psychrobacter arcticus 273-4, com... 168 2e-72 CP000447_3258( CP000447 |pid:none) Shewanella frigid...:none) Acinetobacter baumannii ACICU, ... 169 2e-70 BX640429_39( BX640429 |pid:none) Bordetella parapertussis stra...tella pertussis strain Toha... 149 5e-70 AM902716_2813( AM902716 |pid:none) Bordetella petrii stra

  14. Synergistic Epithelial Responses to Endotoxin and a Naturally Occurring Muramyl Peptide

    Flak, Tod A.; Heiss, Linda N.; Engle, Jacquelyn T.; Goldman, William E


    We have investigated the synergistic interactions of a naturally occurring peptidoglycan fragment (muramyl peptide) and bacterial endotoxin in the induction of inflammatory processes within respiratory epithelial cells, at the levels of both signal transduction events and ultimate cellular metabolic effects. The source of the muramyl peptide is Bordetella pertussis, the causative agent of the respiratory disease pertussis. During log-phase growth, B. pertussis releases the muramyl peptide tra...

  15. The Effects of Adjuvants on Autoimmune Responses Against Testicular Antigens in Mice

    MUSHA, Muhetaerjiang; Hirai, Shuichi; Naito, Munekazu; Terayama, Hayato; Qu, Ning; Hatayama, Naoyuki; Itoh, Masahiro


    Abstract Experimental autoimmune orchitis (EAO) is a model of immunologic male infertility and pathologically characterized by lymphocytic inflammation, which causes breakdown of the testicular immune privilege with spermatogenic disturbance. Generally, murine EAO is induced by immunization with testicular homogenate (TH) from the testes of donor mice + complete Freund's adjuvant (CFA) + Bordetella pertussigens (BP), and it has been considered that treatment with these two adjuvants is requir...

  16. Similarity analysis, synthesis, and bioassay of antibacterial cyclic peptidomimetics

    Workalemahu M. Berhanu


    Full Text Available The chemical similarity of antibacterial cyclic peptides and peptidomimetics was studied in order to identify new promising cyclic scaffolds. A large descriptor space coupled with cluster analysis was employed to digitize known antibacterial structures and to gauge the potential of new peptidomimetic macrocycles, which were conveniently synthesized by acylbenzotriazole methodology. Some of the synthesized compounds were tested against an array of microorganisms and showed antibacterial activity against Bordetella bronchistepica, Micrococcus luteus, and Salmonella typhimurium.

  17. The Lymphocytosis-Promoting Agent Pertussis Toxin Affects Virus Burden and Lymphocyte Distribution in the SIV-Infected Rhesus Macaque

    Pauza, C. David; Hinds, Paul W.; Yin, Cheng; McKechnie, Timothy S.; Hinds, Sarah B; Salvato, Maria S.


    Pertussis toxin from the gram-negative bacterium Bordetella pertussis is an ADP-ribosylase that modifies Gi proteins in mammalian lymphocytes and inhibits their capacity to traffic from blood into lymphoid tissues. We used this compound to induce lymphocytosis in rhesus macaques and to study its effects on SIV infection. Pertussis toxin injected at 25 μg/kg induced a transient lymphocytosis that peaked 3–8 days after administration and caused a rapid, transient decrease in the frequency of in...

  18. Annotating Enzymes of Uncertain Function: The Deacylation of d-Amino Acids by Members of the Amidohydrolase Superfamily†

    Cummings, Jennifer; Fedorov, Alexander A.; Xu, Chengfu; Brown, Shoshana; Fedorov, Elena; Patricia C Babbitt; Almo, Steven C.; Raushel, Frank M.


    The catalytic activities of three members of the amidohydrolase superfamily were discovered using amino acid substrate libraries. Bb3285 from Bordetella bronchiseptica, Gox1177 from Gluconobacter oxydans, and Sco4986 from Streptomyces coelicolor are currently annotated as d-aminoacylases or N-acetyl-d-glutamate deacetylases. These three enzymes are 22−34% identical to one another in amino acid sequence. Substrate libraries containing nearly all combinations of N-formyl-d-Xaa, N-acetyl-d-Xaa, ...

  19. Similarity analysis, synthesis, and bioassay of antibacterial cyclic peptidomimetics

    Berhanu, Workalemahu M; Ibrahim, Mohamed A; Pillai, Girinath G; Oliferenko, Alexander A; Khelashvili, Levan; Jabeen, Farukh; Mirza, Bushra; Ansari, Farzana Latif; ul-Haq, Ihsan; El-Feky, Said A


    Summary The chemical similarity of antibacterial cyclic peptides and peptidomimetics was studied in order to identify new promising cyclic scaffolds. A large descriptor space coupled with cluster analysis was employed to digitize known antibacterial structures and to gauge the potential of new peptidomimetic macrocycles, which were conveniently synthesized by acylbenzotriazole methodology. Some of the synthesized compounds were tested against an array of microorganisms and showed antibacterial activity against Bordetella bronchistepica, Micrococcus luteus, and Salmonella typhimurium. PMID:23019443

  20. Adenylate cyclase toxin-hemolysin relevance for pertussis vaccines

    Šebo, Peter; Osička, Radim; Mašín, Jiří


    Roč. 13, č. 10 (2014), s. 1215-1227. ISSN 1476-0584 R&D Projects: GA ČR GA13-14547S; GA ČR(CZ) GAP302/11/0580; GA ČR GAP302/12/0460 Institutional support: RVO:61388971 Keywords : adenylate cyclase toxin * antigen delivery * Bordetella pertussis Subject RIV: EE - Microbiology, Virology Impact factor: 4.210, year: 2014

  1. Heterosubtypic protection against influenza A induced by adenylate cyclase toxoids delivering conserved HA2 subunit of hemagglutinin

    Staneková, Z.; Adkins, Irena; Kosová, Martina; Janulíková, J.; Šebo, Peter; Varečková, E.


    Roč. 97, č. 1 (2013), s. 24-35. ISSN 0166-3542 R&D Projects: GA ČR GA310/08/0447; GA ČR GP310/09/P582 Institutional support: RVO:61388971 Keywords : Bordetella adenylate cyclase toxoid * Influenza A infection * Cross-protection Subject RIV: FR - Pharmacology ; Medidal Chemistry Impact factor: 3.434, year: 2013

  2. Aerosolized Bacillus anthracis Infection in New Zealand White Rabbits: Natural History and Intravenous Levofloxacin Treatment

    Yee, Steven B.; Hatkin, Joshua M; Dyer, David N; Orr, Steven A.; Pitt, M. Louise M.


    The natural history for inhalational Bacillus anthracis (Ames strain) exposure in New Zealand white rabbits was investigated to better identify potential, early biomarkers of anthrax. Twelve SPF Bordetella-free rabbits were exposed to 150 LD50 aerosolized B. anthracis spores, and clinical signs, body temperature, complete blood count, bacteremia, and presence of protective antigen in the blood (that is, antigenemia) were examined. The development of antigenemia and bacteremia coincided and pr...

  3. Inhibitory effect of the CA2+ antagonist nifedipine on histamine release from rat peritoneal mast cells.



    Full Text Available 45Ca uptake and histamine release was examined in mast cells from rats sensitized with ovalbumin and Bordetella Bertussis as an adjuvant. The uptake of 45Ca by the mast cells was significantly increased by stimulation with ovalbumin as was the release of histamine from the mast cells. Nifedipine, a calcium antagonist, inhibited the increase in both 45Ca uptake and histamine release stimulated by ovalbumin, though the effect on 45Ca uptake was stronger than that on histamine release.

  4. Inhibitory effect of the CA2+ antagonist nifedipine on histamine release from rat peritoneal mast cells.

    Tanizaki,Yoshiro; Komagoe,Haruki; Sudo,Michiyasu; Ohtani,Jun; Kimura,Ikuro; Akagi,Katsumi; Townley, Robert G.


    45Ca uptake and histamine release was examined in mast cells from rats sensitized with ovalbumin and Bordetella Bertussis as an adjuvant. The uptake of 45Ca by the mast cells was significantly increased by stimulation with ovalbumin as was the release of histamine from the mast cells. Nifedipine, a calcium antagonist, inhibited the increase in both 45Ca uptake and histamine release stimulated by ovalbumin, though the effect on 45Ca uptake was stronger than that on histamine release.

  5. Food provisioning alters infection dynamics in populations of a wild rodent.

    Forbes, Kristian M; Henttonen, Heikki; Hirvelä-Koski, Varpu; Kipar, Anja; Mappes, Tapio; Stuart, Peter; Huitu, Otso


    While pathogens are often assumed to limit the growth of wildlife populations, experimental evidence for their effects is rare. A lack of food resources has been suggested to enhance the negative effects of pathogen infection on host populations, but this theory has received little investigation. We conducted a replicated two-factor enclosure experiment, with introduction of the bacterium Bordetella bronchiseptica and food supplementation, to evaluate the individual and interactive effects of pathogen infection and food availability on vole populations during a boreal winter. We show that prior to bacteria introduction, vole populations were limited by food availability. Bordetella bronchiseptica introduction then reduced population growth and abundance, but contrary to predictions, primarily in food supplemented populations. Infection prevalence and pathological changes in vole lungs were most common in food supplemented populations, and are likely to have resulted from increased congregation and bacteria transmission around feeding stations. Bordetella bronchiseptica-infected lungs often showed protozoan co-infection (consistent with Hepatozoon erhardovae), together with more severe inflammatory changes. Using a multidisciplinary approach, this study demonstrates a complex picture of interactions and underlying mechanisms, leading to population-level effects. Our results highlight the potential for food provisioning to markedly influence disease processes in wildlife mammal populations. PMID:26446813

  6. Epidemiology, reemergence of pertussis and vaccine development in Latin America: an overview

    Celso Pérez-Bolaños


    Full Text Available Pertussis o tosferina es una enfermedad bacteriana aguda del tracto respiratorio, causada principalmente por Bordetella pertussis y en menor medida por Bordetella parapertussis. Bordetella pertussis ocupa el quinto lugar en la lista de muertes atribuidas a enfermedades prevenibles por vacunas en niños menores de cinco años en todo el mundo. Se ha reportado que provoca morbilidad y mortalidad significativa, tanto en países desarrollados como en países en desarrollo. La enfermedad es más severa en niños pequeños, pero su prevalencia ha sido observada en todo el mundo en todos los grupos de edad, aún después del desarrollo de vacunas a partir de células completas contra pertussis en los años cuarenta del pasado siglo. Desde la última década ha sido reportada una re-emergencia de pertussis en muchos países desarrollados, incluidos aquellos con una elevada cobertura de vacunación por años. Varios factores pudieran provocar la re-emergencia de pertussis, por ejemplo, una mayor conciencia del problema, un mejor diagnóstico mediante la implementación de técnicas de PCR, disminución de la cobertura de vacunación, la utilización de vacunas de baja protección, baja inmunidad inducida por vacunas y adaptación del patógeno. Este trabajo revisa el estado actual de la epidemiología y la re-emergencia de la enfermedad en América Latina y enfoca la situación actual en Cuba, para dar un panorama de la aplicación de estrategias novedosas en el desarrollo de vacunas futuras, así como medidas generales, recomendadas para el tratamiento de la enfermedad.

  7. Dicty_cDB: Contig-U08273-1 [Dicty_cDB

    Full Text Available as vkfeqnienpfsfkhikilssleelqelpdtnkviltssqdletgfsrelfiqwcsdpkt Frame B: fi*fdginyqiysiiwskr*ittmlfirn**flyiirlwskl*frlfiirt...d:none) Bordetella parapertussis strain ... 59 2e-08 AE008384_695( AE008384 |pid:none) Methanosarcina mazei strain... CR628336 |pid:none) Legionella pneumophila str. Par... 50 4e-06 AY596297_1159( AY596297 |pid:none) Haloarcula marismortui... 58 2e-12 CP000852_446( CP000852 |pid:none) Caldivirga maquilingensis IC-167... 63 2e-12 AP009049_2585( AP00...none) Burkholderia thailandensis E264 ... 46 0.003 CP000387_1483( CP000387 |pid:none) Streptococcus sanguini

  8. Dicty_cDB: Contig-U14924-1 [Dicty_cDB

    Full Text Available hia sp. CCS1, complete g... 603 e-171 CP000140_158( CP000140 |pid:none) Parabacteroides distasonis ATCC ... ...e PB90-1, complet... 446 e-123 (Q8GBW6) RecName: Full=Methylmalonyl-CoA carboxyltransferas...e) Nocardia farcinica IFM 10152 DN... 346 2e-93 A48665( A48665 ) methylmalonyl-CoA carboxyltransferas...e) Burkholderia pseudomallei 668 c... 223 1e-56 CU914168_292( CU914168 |pid:none) Ralstonia solanacearum stra...e) Acidovorax citrulli AAC00-1, co... 220 1e-55 BX640436_177( BX640436 |pid:none) Bordetella parapertussis stra

  9. Forekomst af resistente bakterier og forbrug af antibiotika til hunde

    Pedersen, Karl; Pedersen, Kristina; Jensen, Helene; Finster, Kai; Jensen, Vibeke Frøkjær; Heuer, Ole E.


    Forekomsten af antibiotikaresistens i forskellige patogene bakterier fra hunde blev bestemt og resultaterne sammenholdt med forbrug af antibiotika til hunde i Danmark. I undersøgelsen indgik isolater af Staphylococcus intermedius (n=201), Streptococcus canis (n=37), Pseudomonas aeruginosa (n=39......), Pasteurella multocida (n=25), Bordetella bronchiseptica (n=14), Proteus spp. (n=29), og E. coli (n=449). I undersøgelsen anvendtes data fra VetStat databasen. Størstedelen af de antibiotika, der bruges til hunde er bredspektrede. Penicilliner med udvidet spektrum, cephalosporiner samt sulphonamider...

  10. Epidemiological Aspects of Pertussis among Adults and Adolescents in a Korean Outpatient Setting: A Multicenter, PCR-Based Study

    Park, Sunghoon; Lee, Sun Hwa; Seo, Ki-Hyun; SHIN, KYEONG-CHEOL; Park, Yong Bum; Lee, Myung Goo; Yoo, Kwang Ha; Kim, Hui Jung; Park, Jae Seuk; Cho, Jae Hwa; Ko, Yongchun; Lee, Soo-Keol; Cheon, Ki Tae; Kim, Do Il; Ha, Jun Wook


    Epidemiological data of Bordetella pertussis infection among adolescents and adults are limited in Korea. Patients (≥ 11 yr of age) with a bothersome cough for less than 30 days were enrolled during a 1-yr period at 22 hospitals in Korea. Nasopharyngeal swabs were collected for polymerase chain reaction (PCR) and for bacteriologic culture. In total, 490 patients were finally enrolled, and 34 (6.9%) patients tested positive for B. pertussis; cough duration (14.0 days [7.0-21.0 days]) and age d...

  11. Deciphering Parameter Sensitivity in the BvgAS Signal Transduction.

    Mapder, Tarunendu; Talukder, Srijeeta; Chattopadhyay, Sudip; Banik, Suman K


    To understand the switching of different phenotypic phases of Bordetella pertussis, we propose an optimized mathematical framework for signal transduction through BvgAS two-component system. The response of the network output to the sensory input has been demonstrated in steady state. An analysis in terms of local sensitivity amplification characterizes the nature of the molecular switch. The sensitivity analysis of the model parameters within the framework of various correlation coefficients helps to decipher the contribution of the modular structure in signal propagation. Once classified, the model parameters are tuned to generate the behavior of some novel strains using simulated annealing, a stochastic optimization technique. PMID:26812153

  12. Dicty_cDB: Contig-U13065-1 [Dicty_cDB

    Full Text Available :none) Salmonella enterica subsp. ente... 50 5e-05 CR626927_476( CR626927 |pid:none) Bacteroides fragi...ium chrysogenum Wisconsi... 54 5e-06 AP006841_1248( AP006841 |pid:none) Bacteroides fragilis YC...:none) Ostreococcus lucimarinus CCE9901... 53 1e-05 AM902716_720( AM902716 |pid:none) Bordetella petrii stra...veticus DPC 457... 50 5e-05 CR626927_1013( CR626927 |pid:none) Bacteroides fragilis NCTC 9343,... 50... avium 197N complete ... 50 5e-05 AP006841_1145( AP006841 |pid:none) Bacteroides fragilis YCH46 DNA

  13. Dicty_cDB: Contig-U10300-1 [Dicty_cDB

    Full Text Available ns ORS 2... 116 1e-24 BX640416_10( BX640416 |pid:none) Bordetella pertussis strain Toham... 114 ...:none) Gluconobacter oxydans 621H, com... 93 2e-17 CP000447_3010( CP000447 |pid:none) Shewanella frigid...00447_2198( CP000447 |pid:none) Shewanella frigidimarina NCIMB ... 75 4e-12 CP001103_1911( CP001103 |pid:non...:none) Yersinia pseudotuberculosis strain... 69 2e-10 AE017354_2270( AE017354 |pid:none) Legionella pneumophila...:none) Bacteroides fragilis YCH46 DNA,... 69 3e-10 AY306007_1( AY306007 |pid:none) Da

  14. Dicty_cDB: Contig-U10823-1 [Dicty_cDB

    Full Text Available enterica subsp. enteri... 80 2e-13 CR626927_476( CR626927 |pid:none) Bacteroides fragilis NCTC 9343, ... 7...4e-13 AP006841_551( AP006841 |pid:none) Bacteroides fragilis YCH46 DNA, ... 79 4e-13 BA000023_2698( BA000023 |pid...:none) Saccharopolyspora erythraea NRR... 70 2e-10 AP006841_1248( AP006841 |pid:none) Bacteroides fragi...7 2e-09 AM902716_720( AM902716 |pid:none) Bordetella petrii strain DSM 128... 67 ...tum walsbyi DSM 16790 ... 62 9e-08 CR626927_1013( CR626927 |pid:none) Bacteroides fragilis NCTC 9343,...

  15. Dicty_cDB: Contig-U15213-1 [Dicty_cDB

    Full Text Available rochlorococcus marinus str. NAT... 95 4e-18 CP000937_1559( CP000937 |pid:none) Francisella philomiragi...:none) Methylobacterium nodulans ORS 2... 88 4e-16 AP006841_3263( AP006841 |pid:none) Bacteroides fragili...:none) Shewanella denitrificans OS217,... 105 4e-21 CP000115_205( CP000115 |pid:none) Nitrobacter winogra...:none) Lactobacillus gasseri ATCC 33323... 100 9e-20 AE014299_2458( AE014299 |pid:none) Shewanella oneide... sp. JS666, complete... 99 3e-19 AM902716_1733( AM902716 |pid:none) Bordetella petrii strain DSM 12... 99 3e

  16. Dicty_cDB: Contig-U13788-1 [Dicty_cDB

    Full Text Available id ... 44 0.001 AY466441_16( AY466441 |pid:none) Saccharopolyspora spinosa NRLL 1...32... 108 4e-23 AE017343_266( AE017343 |pid:none) Cryptococcus neoformans var. neo... 108 4e-23 CU928179_8( ...4 AE010299_2239( AE010299 |pid:none) Methanosarcina acetivorans str.... 77 2e-13 BX294147_33( BX294147 |pid:...001195 |pid:none) Rhizobium leguminosarum bv. trif... 42 0.009 CP000159_172( CP00...none) Bordetella avium 197N complete ... 38 0.096 AE010299_4447( AE010299 |pid:none) Methanosarcina acetivor

  17. Triazenos e atividade antibacteriana Triazenes and antibacterial activity

    Manfredo Hörner; Vinícius Feltrin Giglio; Aline Joana Rolina Wohlmuth Alves dos Santos; André Bilibio Westphalen; Bernardo Almeida Iglesias; Paulo Roberto Martins; Carlos Henrique do Amaral; Tiago Mozaquatro Michelot; Luiz Gustavo Brenner Reetz; Cláudia de Mello Bertoncheli; Gustavo Luiz Paraginski; Rosmari Horner


    Quinze compostos triazenos foram estudados quanto à atividade antibacteriana pela metodologia de microdiluição em caldo. A Concentração Inibitória Mínima (CIM) e a Concentração Bactericida Mínima (CBM) foram pesquisadas frente a três bactérias padrão (E. coli ATCC 25922, Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 27853) e frente a cepas hospitalares (Acinetobacter baumannii, Acinetobacter lwoffii, Ralstonia pickettii, Bordetella bronchiseptica, Micrococcus sp., Enterococcus...

  18. BtcA, A class IA type III chaperone, interacts with the BteA N-terminal domain through a globular/non-globular mechanism.

    Chen Guttman

    Full Text Available Bordetella pertussis, the etiological agent of "whooping cough" disease, utilizes the type III secretion system (T3SS to deliver a 69 kDa cytotoxic effector protein, BteA, directly into the host cells. As with other T3SS effectors, prior to its secretion BteA binds BtcA, a 13.9 kDa protein predicted to act as a T3SS class IA chaperone. While this interaction had been characterized for such effector-chaperone pairs in other pathogens, it has yet to be fully investigated in Bordetella. Here we provide the first biochemical proof that BtcA is indeed a class IA chaperone, responsible for the binding of BteA's N-terminal domain. We bring forth extensive evidence that BtcA binds its substrate effector through a dual-interface binding mechanism comprising of non-globular and bi-globular interactions at a moderate micromolar level binding affinity. We demonstrate that the non-globular interactions involve the first 31 N-terminal residues of BteA287 and their removal leads to destabilization of the effector-chaperone complex and lower binding affinities to BtcA. These findings represent an important first step towards a molecular understanding of BteA secretion and cell entry.

  19. Avaliação sorológica de vacinações preventivas da difteria, do tétano e da coqueluche, efetuadas em crianças prematuras

    Vicente Amato Neto


    Full Text Available Vinte crianças prematuras receberam, no primeiro ano de vida. vacinas que habitualmente fazem parte do esquema básico de imunizações ativas. Em amostra de soro obtida quando elas atingiram a idade de 12 meses, foram dosados os teores de antitoxina diftérica, de antitoxina tetânica e de aglutininas anti Bordetella pertussis. Valores plenamente satisfatórios de anticorpos relativos à difteria e ao tétano puderam ser encontrados e, quanto à coqueluche, nunca notaram os Autores ausência de aglutininas, mas conclusão mais decisiva não ocorreu, em virtude da falta de melhor conhecimento da cifra indicativa de proteção. O estudd em questão representa subsídio no sentido de arrefecer o temor e o cepticismo, bastante divulgados, acerca da vacinação de prematuros.Twenty premature-born children received, during their first year of life, vaccines routinely apptied as part of a basic immunization schedule. Sera obtained at the age of 12 months were titered for antibodies against diphteria, tetanus and pertussis. Values considered protective were observed for diphteria and tetanus. Anti - Bordetella pertussis agglutinins were always present, however, in the absence of a consensus as to what are protective levels, no conclusion could be drawn. The present study contributes towards erasing the prejudice and scepticism concerning the immunization of the premature-born.

  20. Genome implosion elicits host-confinement in Alcaligenaceae: evidence from the comparative genomics of Tetrathiobacter kashmirensis, a pathogen in the making.

    Wriddhiman Ghosh

    Full Text Available This study elucidates the genomic basis of the evolution of pathogens alongside free-living organisms within the family Alcaligenaceae of Betaproteobacteria. Towards that end, the complete genome sequence of the sulfur-chemolithoautotroph Tetrathiobacter kashmirensis WT001(T was determined and compared with the soil isolate Achromobacter xylosoxidans A8 and the two pathogens Bordetella bronchiseptica RB50 and Taylorella equigenitalis MCE9. All analyses comprehensively indicated that the RB50 and MCE9 genomes were almost the subsets of A8 and WT001(T, respectively. In the immediate evolutionary past Achromobacter and Bordetella shared a common ancestor, which was distinct from the other contemporary stock that gave rise to Tetrathiobacter and Taylorella. The Achromobacter-Bordetella precursor, after diverging from the family ancestor, evolved through extensive genome inflation, subsequent to which the two genera separated via differential gene losses and acquisitions. Tetrathiobacter, meanwhile, retained the core characteristics of the family ancestor, and Taylorella underwent massive genome degeneration to reach an evolutionary dead-end. Interestingly, the WT001(T genome, despite its conserved architecture, had only 85% coding density, besides which 578 out of its 4452 protein-coding sequences were found to be pseudogenized. Translational impairment of several DNA repair-recombination genes in the first place seemed to have ushered the rampant and indiscriminate frame-shift mutations across the WT001(T genome. Presumably, this strain has just come out of a recent evolutionary bottleneck, representing a unique transition state where genome self-degeneration has started comprehensively but selective host-confinement has not yet set in. In the light of this evolutionary link, host-adaptation of Taylorella clearly appears to be the aftereffect of genome implosion in another member of the same bottleneck. Remarkably again, potent virulence factors

  1. Cough and dyspnoea of an asthmatic patient at Mt. Kilimanjaro: a difficult differential diagnosis.

    Goebbels, K; Gieseler, U; Schöffl, Volker; Küpper, Thomas


    This case highlights the difficulties associated with the differential diagnosis of pulmonary symptoms in patients with pre-existing diseases in extreme environmental conditions. A 58-year-old man with child-onset allergic asthma developed dyspnoea and an acute non-productive cough during a trekking expedition on Mt. Kilimanjaro (5895m) in Tanzania. The symptoms were believed initially to be linked to the high altitude exposure (high altitude pulmonary oedema (HAPE) or high altitude cough) or his pre-existing asthma. However, he was later diagnosed correctly with a reinfection of Bordetella pertussis. Pertussis is a highly communicable disease with potentially serious medical consequences that could have affected all of the expedition members. The effectiveness of a pertussis vaccine declines 4-12 years after the vaccination. Thus, it is suggested that the status of immunisation against pertussis should be checked along with those of other infections prior to travel. PMID:20188301

  2. Dicty_cDB: VHO891 [Dicty_cDB

    Full Text Available ces producing significant alignments: (bits) Value (Q54X49) RecName: Full=5-methyltetrahydropteroyltriglutama...te--ho... 479 e-134 (Q2KYF3) RecName: Full=5-methyltetrahydropteroyltriglutamate...--ho... 308 2e-82 (Q7W791) RecName: Full=5-methyltetrahydropteroyltriglutamate--ho... 304 2e-81 BX640431_54(... BX640431 |pid:none) Bordetella parapertussis strain 1... 304 2e-81 (Q7VVU3) RecName: Full=5-methyltetrahydropteroyltriglutama...te--ho... 304 2e-81 (A9N9C7) RecName: Full=5-methyltetrahydropteroyltriglutamate--ho... 303 4e-81 (B6J3R8) RecNam

  3. Dicty_cDB: Contig-U00515-1 [Dicty_cDB

    Full Text Available ) Cryptococcus neoformans var. neo... 43 0.029 AY194224_1( AY194224 |pid:none) Aedes aegypti strain...a genomic DNA ch... 55 6e-06 BX640431_139( BX640431 |pid:none) Bordetella parapertussis strain...8_635( CU914168 |pid:none) Ralstonia solanacearum strain IP... 39 0.33 CP000687_334( CP000687 |pid:none) Actinobacillus pleuropne.... Toulou... 39 0.56 AJ496288_1( AJ496288 |pid:none) Bartonella tribocorum succinate de... 39 0.56 (Q7S3C9) RecName: Full=Kynurenine...) Ralstonia solanacearum strain Mo... 37 1.2 CP000750_2785( CP000750 |pid:none) Kine

  4. Dicty_cDB: Contig-U12088-1 [Dicty_cDB

    Full Text Available 7 G02257( G02257 ) NAD(P) transhydrogenase (B-specific) (EC 424 e-117 AM920436_1880( AM920436 |p...... 417 e-114 S54876( S54876 ) NAD(P) transhydrogenase (B-specific) (EC 414 e-114 AK088044_...enase (B-specific) (EC... 296 3e-78 AM421808_905( AM421808 |pid:none) Neisseria meningitidis serogro...44( CP000510 |pid:none) Psychromonas ingrahamii 37, comp... 280 2e-73 CP000115_989( CP000115 |pid:none) Nitrobacter winogra...6 3e-72 BX640430_209( BX640430 |pid:none) Bordetella parapertussis strain ... 275 5e-72 AB3550( AB3550 ) NAD(P) transhydrog

  5. Dicty_cDB: Contig-U16436-1 [Dicty_cDB

    Full Text Available 5 e-114 CU633870_115( CU633870 |pid:none) Podospora anserina genomic DNA c... 344 e-114 AJ621287_1( AJ621287...015928 |pid:none) Bacteroides thetaiotaomicron VP... 39 0.90 CP000099_229( CP000099 |pid:none) Methanosarcina...nkkkkkkgrppp Translated Amino Acid sequence (All Frames) Frame A: LIFFFLININKFIIIEIKIFYFL...098096_5( EU098096 |pid:none) Blumeria graminis f. sp. hordei Av... 325 2e-99 A67854_1( A67854 |pid:none) Sequence 26 from Paten...0615_1648( CP000615 |pid:none) Burkholderia vietnamiensis G4 c... 271 8e-71 AM902716_277( AM902716 |pid:none) Bordetella petrii stra

  6. Dicty_cDB: Contig-U14087-1 [Dicty_cDB

    Full Text Available total letters Score E Sequences producing significant alignments: (bits) Value Contig-U14087-1 (Contig-U14087-1Q) /CSM_Contig/ S-transferase Y-b subu... 55 2e-06 (Q9JHF7) RecName: Full=Glutathione-requiring prostaglandin D Bordetella petrii strain DSM 12... 54 6e-06 ( O73888 ) RecName: Full=Glutathione-requiring prostaglandin...( AX886166 |pid:none) Sequence 2029 from Patent EP1033401. 49 3e-04 DQ789057_1( DQ789057 |pid:none) Ciona intestinalis prostaglandin... WO20080... 46 0.002 DQ789056_1( DQ789056 |pid:none) Ciona intestinalis prostaglandin D... 46 0

  7. Disodium cromoglycate inhibits production of immunoglobulin E.

    Seo, S B; Park, S J; Park, S T; Cho, C C; Park, B H; Lee, S J; Kim, H M; Kajiuchi, T; Shin, T Y


    Disodium cromoglycate (DSCG) has been shown to inhibit the release of mediators from mast cells. In the present study, the effect of DSCG on active anaphylactic reaction was studied in mice. DSCG dose-dependently inhibited the active systemic anaphylactic reaction and serum immunoglobulin (Ig)E production induced by immunization with ovalbumin, Bordetella pertussis toxin and aluminum hydroxide gel. DSCG strongly inhibited IL-4-dependent IgE production by lipopolysaccharide-stimulated murine whole spleen cells. In the case of U266 human IgE-bearing B cells, DSCG also showed an inhibitory effect on the IgE production. These results suggest that DSCG has an anti-anaphylactic activity by inhibition of IgE production from B cells. PMID:11417850

  8. Pertussis in young infants: clinical presentation, course and prevention.

    O'Riordan, A; Cleary, J; Cunney, R; Nicholson, A J


    Pertussis is a highly contagious disease caused by the Gram negative aerobic coccobacillus, Bordetella pertussis. It may present with severe symptoms and complications in infants and can pose a diagnostic challenge. This is a vaccine preventable illness covered by the Irish Childhood Immunisation Schedule. In 2011, a retrospective review was conducted of the records of infants, under six months, with a confirmed diagnosis of pertussis, presenting to Temple Street Children's University Hospital (TSCUH). A summery of notifications of pertussis nationally, from 2001 to 2012, was also examined as part of the study. This found that the rate of reported cases of pertussis has been increasing in Ireland. This national increase corresponds with a rising number of cases identified at TSCUH. Patients commonly presented severely ill with cyanosis and apnoea, on a background of prolonged cough. We found that pertussis was diagnosed rapidly in most cases however in all cases there was a delay to commencement of appropriate macrolide therapy. PMID:25226721

  9. AcEST: BP918529 [AcEST

    Full Text Available 29p OS=Yarrowia lipolytica GN=YA... 33 9.8 tr|Q2UKB0|Q2UKB0_ASPOR Histone acetyltransferase...YMU001_000114_E12. (549 letters) Database: uniprot_sprot.fasta 412,525 sequences; 148,809,765 total letters ...phosphoglycerate-dependent phosphoglycerate mutase OS=Bordetella bronchiseptica GN=gpmA PE=3 SV=1 Length = 2...DDIVGVNIPTGQP 211 >sp|Q9JI69|ORC1_CRIGR Origin recognition complex subunit 1 OS=Cricetulus griseus GN=ORC1L ... YMU001_000114_E12. (549 letters) Database: uniprot_trembl.fasta 7,341,751 sequences; 2,391,615,440 total le

  10. Dicty_cDB: Contig-U13365-1 [Dicty_cDB

    Full Text Available 666( CU928168 |pid:none) Kluyveromyces thermotolerans str... 41 0.015 FM992695_586( FM992695 |pid:none) Candida dubliniens...28168_242( CU928168 |pid:none) Kluyveromyces thermotolerans str... 83 3e-15 CR382130_959( CR382130 |pid:none...agilis YCH46 DNA,... 50 3e-05 CU928167_24( CU928167 |pid:none) Kluyveromyces thermotolerans stra... 49 4e-05...cillus cereus subsp. cytotoxi... 32 6.9 AB209036_1( AB209036 |pid:none) Homo sapiens mRNA for prot...34 1.4 AM902716_4268( AM902716 |pid:none) Bordetella petrii strain DSM 12... 34 1.4 BX293980_487( BX293980 |pid:none) Mycoplasma myco

  11. Dicty_cDB: Contig-U12565-1 [Dicty_cDB

    Full Text Available l... 360 1e-97 FN392320_765( FN392320 |pid:none) Pichia pastoris GS115 chromo...5 3 ( EJ802965 ) 1093017441017 Global-Ocean-Sampling_GS-30-02-01-1... 46 3e-05 3 ( CP000236 ) Ehrlichia chaffeensis str. Arkansas...0.001 1 ( CR767821 ) Ehrlichia ruminantium strain Welgevonden, complet... 58 0.001 1 ( AC178441 ) Strongyloc...S9; se... 221 1e-55 CU928164_4106( CU928164 |pid:none) Escherichia coli IAI39 chromoso... 221 1e-55 (P0AB77)...M167904 |pid:none) Bordetella avium 197N complete ... 185 6e-45 CU928145_796( CU928145 |pid:none) Escherichia coli 55989 chromo

  12. Dark fermentative hydrogen production by defined mixed microbial cultures immobilized on ligno-cellulosic waste materials

    Patel, Sanjay K.S. [Microbial Biotechnology and Genomics, Institute of Genomics and Integrative Biology (IGIB), CSIR, Delhi University Campus, Mall Road, Delhi 110007 (India); Department of Biotechnology, University of Pune, Pune 411007 (India); Purohit, Hemant J. [Environmental Genomics Unit, National Environmental Engineering Research Institute (NEERI), CSIR, Nehru Marg, Nagpur 440020 (India); Kalia, Vipin C. [Microbial Biotechnology and Genomics, Institute of Genomics and Integrative Biology (IGIB), CSIR, Delhi University Campus, Mall Road, Delhi 110007 (India)


    Mixed microbial cultures (MMCs) based on 11 isolates belonging to Bacillus spp. (Firmicutes), Bordetella avium, Enterobacter aerogenes and Proteus mirabilis (Proteobacteria) were employed to produce hydrogen (H{sub 2}) under dark fermentative conditions. Under daily fed culture conditions (hydraulic retention time of 2 days), MMC6 and MMC4, immobilized on ligno-cellulosic wastes - banana leaves and coconut coir evolved 300-330 mL H{sub 2}/day. Here, H{sub 2} constituted 58-62% of the total biogas evolved. It amounted to a H{sub 2} yield of 1.54-1.65 mol/mol glucose utilized over a period of 60 days of fermentation. The involvement of various Bacillus spp. -Bacillus sp., Bacillus cereus, Bacillus megaterium, Bacillus pumilus and Bacillus thuringiensis as components of the defined MMCs for H{sub 2} production has been reported here for the first time. (author)

  13. Dicty_cDB: Contig-U12542-1 [Dicty_cDB

    Full Text Available _2922( CP000891 |pid:none) Shewanella baltica OS195, compl... 58 9e-07 CP000115_313( CP000115 |pid:none) Nitrobacter winogra...plet... 57 1e-06 BC161525_1( BC161525 |pid:none) Xenopus tropicalis hypothetical pr... 57 1e-06 CP000259_135...e-06 BX640437_205( BX640437 |pid:none) Bordetella bronchiseptica strain... 56 3e-06 (Q73EU1) RecName: Full=D...IA... 56 3e-06 CP000090_896( CP000090 |pid:none) Ralstonia eutropha JMP134 chromo... 56 3e-06 CU928174_244( DEAD box helicase p... 50 0.22 1 ( AY070052 ) Arabidopsis thaliana putati

  14. Dicty_cDB: Contig-U12487-1 [Dicty_cDB

    Full Text Available . 4b F... 87 1e-15 CP000232_1858( CP000232 |pid:none) Moorella thermoacetica ATCC 390... 87 2e-15 (Q03AH0...mple... 83 3e-14 BX640444_79( BX640444 |pid:none) Bordetella bronchiseptica strain ... 83 3e-14 AM778892_7( AM778892 |pid:none) SB, c... 81 1e-13 CP000232_1860( CP000232 |pid:none) Moorella thermoacetica ATCC 3...s 2336, complet... 81 1e-13 CR382131_686( CR382131 |pid:none) Yarrowia lipolytica strain CLIB1... 81 1... c... 76 2e-12 AM260479_419( AM260479 |pid:none) Ralstonia eutropha H16 chromosom

  15. Dicty_cDB: Contig-U13863-1 [Dicty_cDB

    Full Text Available :none) Francisella philomiragia subsp. ... 49 2e-07 CP001107_52( CP001107 |pid..... 48 0.35 2 ( AC116305 ) Dictyostelium discoideum chromosome 2 map 1005175... 34 0.37 10 ( AP009180 ) Candidatus Carsonella...e-07 ( P42975 ) RecName: Full=Bifunctional protein birA; Includes: Re... 44 2e-07 AE014299_212( AE014299 |pid:none) Shewanella oneide... 43 8e-06 CP000447_132( CP000447 |pid:none) Shewanella frigidimarina NCIMB 4... 44 8e-06 CP000559_54( CP000559 |pid...AM902716_4830( AM902716 |pid:none) Bordetella petrii strain DSM 12... 50 1e-04 CP

  16. Dicty_cDB: Contig-U08344-1 [Dicty_cDB

    Full Text Available 001 3 ( EF452805 ) Rasbora daniconius NADH dehydrogenase subunit 4 (... 56 0.001 1 ( BN001180 ) TPA: Hydra magnipapillata mitochondr...aplotype ... 79 6e-14 CP000524_743( CP000524 |pid:none) Bartonella bacilliformis KC583, ... 68 7e-14 T13881( T13881 ) NADH2 dehydr...s taitungensis mitochondrial ... 51 3e-12 BX640433_157( BX640433 |pid:none) Bordetella parapertussis strain ...64 5e-12 AY832174_1( AY832174 |pid:none) Amborella trichopoda NADH dehydrog... 60 5e-12 CP000767_146( CP000767 |pid...:none) Candidatus Hamiltonella defensa... 57 8e-12 CP001130_749( CP001130 |pid:none) Hydrogen

  17. Dicty_cDB: Contig-U10246-1 [Dicty_cDB

    Full Text Available :none) Nocardioides sp. JS614, complet... 174 6e-42 CP000937_1433( CP000937 |pid:none) Francisella philomiragi...37. 40 1e-08 4 ( AL419645 ) T3 end of clone XAX0AA002A05 of library XAX0AA fr... 48 2e-05 2 ( CP000447 ) Shewanella frigid...hromos... 177 4e-43 AE014299_851( AE014299 |pid:none) Shewanella oneidensis MR-1, comp... 177 4e-43 CP000109_626( CP000109 |pid...:none) Shewanella frigidimarina NCIMB 4... 176 1e-42 CP000789_3430( CP000789 |pid:none) Vibrio ...640411_154( BX640411 |pid:none) Bordetella pertussis strain Toha... 141 4e-32 CP000449_150( CP000449 |pid:no

  18. Dicty_cDB: Contig-U14398-1 [Dicty_cDB

    Full Text Available mRNA for Pl10-relate... 126 4e-28 CP001359_3124( CP001359 |pid:none) Anaeromyxobacter dehalogenan...rica EST, 5' end sequence, clone ... 62 5e-09 2 ( CU432015 ) Clytia hemisphaerica 5-PRIME EST from clo...X640420 |pid:none) Bordetella pertussis strain Toha... 106 2e-29 CP000378_1692( CP000378 |pid:none) Burkholderia cenocep...4_905( CP000544 |pid:none) Halorhodospira halophila SL1, co... 110 2e-31 BT066681_1( BT066681 |pid:none) Zea...teromonas haloplanktis ... 112 7e-31 CP001277_275( CP001277 |pid:none) Candidatus Hamiltonella

  19. Expansion of space station diagnostic capability to include serological identification of viral and bacterial infections

    Hejtmancik, Kelly E.


    It is necessary that an adequate microbiology capability be provided as part of the Health Maintenance Facility (HMF) to support expected microbial disease events during long periods of space flight. The applications of morphological and biochemical studies to confirm the presence of certain bacterial and fungal disease agents are currently available and under consideration. This confirmation would be greatly facilitated through employment of serological methods to aid in the identification for not only bacterial and fungal agents, but viruses as well. A number of serological approached were considered, particularly the use of Enzyme Linked Immunosorbent Assays (ELISAs), which could be utilized during space flight conditions. A solid phase, membrane supported ELISA for the detection of Bordetella pertussis was developed to show a potential model system that would meet the HMF requirements and specifications for the future space station. A second model system for the detection of Legionella pneumophilia, an expected bacterial disease agent, is currently under investigation.

  20. Carrot cells: a pioneering platform for biopharmaceuticals production.

    Rosales-Mendoza, Sergio; Tello-Olea, Marlene Anahí


    Carrot (Daucus carota L.) is of importance in the molecular farming field as it constitutes the first plant species approved to produce biopharmaceuticals for human use. In this review, features that make carrot an advantageous species in the molecular farming field are analyzed and a description of the developments achieved with this crop thus far is presented. A guide for genetic transformation procedures is also included. The state of the art comprises ten vaccine prototypes against Measles virus, Hepatitis B virus, Human immunodeficiency virus, Yersinia pestis, Chlamydia trachomatis, Mycobacterium tuberculosis, enterotoxigenic Escherichia coli, Corynebacterium diphtheria/Clostridium tetani/Bordetella pertussis, and Helicobacter pylori; as well as the case of the glucocerebrosidase, an enzyme used for replacement therapy, and other therapeutics. Perspectives for these developments are envisioned and innovations are proposed such as the use of transplastomic technologies-, hairy roots-, and viral expression-based systems to improve yields and develop new products derived from this advantageous plant species. PMID:25572939

  1. Tracking Pertussis and Evaluating Control Measures through Enhanced Pertussis Surveillance, Emerging Infections Program, United States.

    Skoff, Tami H; Baumbach, Joan; Cieslak, Paul R


    Despite high coverage with pertussis-containing vaccines, pertussis remains endemic to the United States. There have been increases in reported cases in recent years, punctuated by striking epidemics and shifting epidemiology, both of which raise questions about current policies regarding its prevention and control. Limited data on pertussis reported through the National Notifiable Disease Surveillance System have proved insufficient to answer these questions. To address shortcomings of national pertussis data, the Emerging Infections Program at the US Centers for Disease Control and Prevention launched Enhanced Pertussis Surveillance (EPS), which is characterized by systematic case ascertainment, augmented data collection, and collection of Bordetella pertussis isolates. Data collected through EPS have been instrumental in understanding the rapidly evolving epidemiology and molecular epidemiology of pertussis and have contributed essential information regarding pertussis vaccines. EPS also serves as a platform for conducting critical and timely evaluations of pertussis prevention and control strategies, including targeting of vaccinations and antimicrobial prophylaxis. PMID:26291475

  2. Dicty_cDB: Contig-U00735-1 [Dicty_cDB

    Full Text Available equence 11 from Patent WO03040681. 49 2e-04 CU207211_886( CU207211 |pid:none) Herminiimonas arsenicoxydans c..., clone: mib07054, 3' end, expre... 44 6.7 1 ( BG148484 ) uu79b10.y1 Soares_mouse_NMGB_bcell Mus musculus c....IK... 44 6.7 1 ( AA276535 ) vc44g11.r1 Soares mouse 3NbMS Mus musculus cDNA c... 44 6.7 1 ( AA184659 ) mt58d08.r1 Soares_thymus...ulfovibrio desulfuricans sub... 229 4e-59 AP010656_521( AP010656 |pid:none) Candidatus Azobacteroides pse..._1499( AP009384 |pid:none) Azorhizobium caulinodans ORS 57... 56 1e-06 BX640415_270( BX640415 |pid:none) Bordetella pertus

  3. Dicty_cDB: Contig-U06930-1 [Dicty_cDB

    Full Text Available 5e-36 BX640424_184( BX640424 |pid:none) Bordetella parapertussis strain ... 121 2e-26 AM270285_2( AM270285 |pid:none) Asper...gkiiikiiiiii*kckk*nyqyhhliilkmqlkklewqkmhgiqgmqklfhlht vwiqngeigmnllmvehkllnf*krngkkrlnid*lkscghfklvvielpsdlhmngki* mvigldhmemkig...P001117_8( CP001117 |pid:none) Deinococcus deserti VCD115 plasmid... 191 2e-47 CP000580_5560( CP000580 |pid:none) Mycobacter...YN57) RecName: Full=DNA-directed RNA polymerase subunit beta;... 33 5.9 CP000903_542( CP000903 |pid:none) Bacillus wei...llate St... 42 5e-04 2 ( AZ523555 ) 221PbA02 Pb MBN #21 Plasmodium berghei genomic 3'.

  4. Dicty_cDB: Contig-U15022-1 [Dicty_cDB

    Full Text Available la DC2201 DNA, ... 124 2e-51 AP006618_632( AP006618 |pid:none) Nocardia farcinica IFM 10152 DNA... 115 2e-51...ite gut library Reticulitermes flav... 48 0.005 2 ( BC051239 ) Xenopus laevis hypothetical protein...ia enterocolitica subsp. ... 119 6e-54 BX640422_257( BX640422 |pid:none) Bordetella pertussis strain...00096_1320( CP000096 |pid:none) Pelodictyon luteolum DSM 273, c... 128 3e-48 CP000521_3687( CP000521 |pid:none) Acinetobacter bauma...) Pseudomonas aeruginosa PA7, com... 111 5e-46 CP000089_3304( CP000089 |pid:none) Dechloromonas aromatica RCB, co

  5. Functional coupling between heterologously expressed dopamine D(2) receptors and KCNQ channels

    Ljungstrom, Trine; Grunnet, Morten; Jensen, Bo Skaaning;


    Activation of KCNQ potassium channels by stimulation of co-expressed dopamine D(2) receptors was studied electrophysiologically in Xenopus laevis oocytes and in mammalian cells. To address the specificity of the interaction between D(2)-like receptors and KCNQ channels, combinations of KCNQ1......-5 channels and D(2)-like receptors (D(2L), D(3), and D(4)) were investigated in Xenopus oocytes. Activation of either receptor with the selective D(2)-like receptor agonist quinpirole (100 nM) stimulated all the KCNQ currents, independently of the subunit combination, indicating a common pathway of receptor......-channel interaction. The KCNQ4 current was investigated in further detail and was increased by 19.9+/-1.6% ( n=20) by D(2L) receptor stimulation. The effect could be mimicked by injection of GTPgammaS and prevented by injection of Bordetella pertussis toxin, indicating that channel stimulation was mediated via a G...

  6. Dicty_cDB: Contig-U11939-1 [Dicty_cDB

    Full Text Available ffktirg*ylkkkkkkkk*q*ircqhryg nfckikkkk Frame B: ftinkqkhkl*yyyekhifflff*nllkkiivff*kkkkk*nwkmkikignilknvvilv ifslfiskinsevvkpnpak...7.7 AB376036_1( AB376036 |pid:none) Tanakia lanceolata V2R gene for vo... 35 7.7 AM167904_1899( AM167904 |pid:none) Bordetella...cant alignments: (bits) Value N ( BJ424967 ) Dictyostelium discoideum cDNA clone:ddv54n05, 5' ...... 1185 0.0 1 ( DQ447637 ) Dictyostelium discoideum GluPR (GluPR) mRNA, comp... 936 0.0 2 ( BJ440362 ) Dictyostelium cDNA clone:ddv43o09, 3' ... 922 0.0 2 ( BJ438653 ) Dictyostelium discoide

  7. Avaliação sorológica de vacinações preventivas da difteria, do tétano e da coqueluche, efetuadas em crianças prematuras

    Vicente Amato Neto


    Full Text Available Vinte crianças prematuras receberam, no primeiro ano de vida. vacinas que habitualmente fazem parte do esquema básico de imunizações ativas. Em amostra de soro obtida quando elas atingiram a idade de 12 meses, foram dosados os teores de antitoxina diftérica, de antitoxina tetânica e de aglutininas anti Bordetella pertussis. Valores plenamente satisfatórios de anticorpos relativos à difteria e ao tétano puderam ser encontrados e, quanto à coqueluche, nunca notaram os Autores ausência de aglutininas, mas conclusão mais decisiva não ocorreu, em virtude da falta de melhor conhecimento da cifra indicativa de proteção. O estudd em questão representa subsídio no sentido de arrefecer o temor e o cepticismo, bastante divulgados, acerca da vacinação de prematuros.

  8. AcEST: BP915876 [AcEST

    Full Text Available QPAGKTI 271 L+P +T+ Sbjct: 878 HLEPKFRTL 886 >sp|Q08530|BTR_BORPE Transcriptional regulatory protein btr OS=Bordetella pertussis...-1 OS=Bos taurus GN=DSC1 PE=1 SV=1 32 0.99 sp|Q08530|BTR_BORPE Transcriptional regulatory protein btr OS=Bo... 31 1.7 sp...otein database search programs, Nucleic Acids Res. 25:3389-3402. Query= BP915876|Adiantum capillus-veneris...YMU001_000079_A02 337 Adiantum capillus-veneris mRNA. clone: YMU001_000079_A02. BP9...15876 - Show BP915876 Clone id YMU001_000079_A02 Library YMU01 Length 337 Definition Adiantum capillus-veneris

  9. Multi-functional characteristics of the Pseudomonas aeruginosa type III needle-tip protein, PcrV; comparison to orthologs in other gram negative bacteria

    Hiromi eSato


    Full Text Available Pseudomonas aeruginosa possesses a type III secretion system (T3SS to intoxicate host cells and evade innate immunity. This virulence-related machinery consists of a molecular syringe and needle assembled on the bacterial surface, which allows delivery of T3 effector proteins into infected cells. To accomplish a one-step effector translocation, a tip protein is required at the top end of the T3 needle structure. Strains lacking expression of the functional tip protein fail to intoxicate host cells.P. aeruginosa encodes a T3S that is highly homologous to the proteins encoded by Yersinia species. The needle tip proteins of Yersinia, LcrV, and P. aeruginosa, PcrV, share 37% identity and 65% similarity. Other known tip proteins are AcrV (Aeromonas, IpaD (Shigella, SipD (Salmonella, BipD (Burkholderia, EspA (EPEC, EHEC, Bsp22 (Bordetella, with additional proteins identified from various Gram negative species, such as Vibrio and Bordetella. The tip proteins can serve as a protective antigen or may be critical for sensing host cells and evading innate immune responses. Recognition of the host microenvironment transcriptionally activates synthesis of T3SS components. The machinery appears to be mechanically controlled by the assemblage of specific junctions within the apparatus. These junctions include the tip and base of the T3 apparatus, the needle proteins and components within the bacterial cytoplasm. The tip proteins likely have chaperone functions for translocon proteins, allowing the proper assembly of translocation channels in the host membrane and completing vectorial delivery of effector proteins into the host cytoplasm. Multifunctional features of the needle-tip proteins appear to be intricately controlled. In this review, we highlight the functional aspects and complex controls of T3 needle-tip proteins with particular emphasis on PcrV and LcrV.

  10. Molecular Structure of WlbB, a Bacterial N-Acetyltransferase Involved in the Biosynthesis of 2,3-Diacetamido-2,3-dideoxy-d-mannuronic Acid

    Thoden, James B.; Holden, Hazel M. (UW)


    The pathogenic bacteria Pseudomonas aeruginosa and Bordetella pertussis contain in their outer membranes the rare sugar 2,3-diacetamido-2,3-dideoxy-D-mannuronic acid. Five enzymes are required for the biosynthesis of this sugar starting from UDP-N-acetylglucosamine. One of these, referred to as WlbB, is an N-acetyltransferase that converts UDP-2-acetamido-3-amino-2,3-dideoxy-D-glucuronic acid (UDP-GlcNAc3NA) to UDP-2,3-diacetamido-2,3-dideoxy-D-glucuronic acid (UDP-GlcNAc3NAcA). Here we report the three-dimensional structure of WlbB from Bordetella petrii. For this analysis, two ternary structures were determined to 1.43 {angstrom} resolution: one in which the protein was complexed with acetyl-CoA and UDP and the second in which the protein contained bound CoA and UDP-GlcNAc3NA. WlbB adopts a trimeric quaternary structure and belongs to the L{beta}H superfamily of N-acyltransferases. Each subunit contains 27 {beta}-strands, 23 of which form the canonical left-handed {beta}-helix. There are only two hydrogen bonds that occur between the protein and the GlcNAc3NA moiety, one between O{sup {delta}1} of Asn 84 and the sugar C-3{prime} amino group and the second between the backbone amide group of Arg 94 and the sugar C-5{prime} carboxylate. The sugar C-3{prime} amino group is ideally positioned in the active site to attack the si face of acetyl-CoA. Given that there are no protein side chains that can function as general bases within the GlcNAc3NA binding pocket, a reaction mechanism is proposed for WlbB whereby the sulfur of CoA ultimately functions as the proton acceptor required for catalysis.