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Sample records for bordetella

  1. Characterization of fimbrial subunits from Bordetella species

    NARCIS (Netherlands)

    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.

    1987-01-01

    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.

    Science.gov (United States)

    Hulbert, Robin R; Cotter, Peggy A

    2009-11-01

    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

    Science.gov (United States)

    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

    DEFF Research Database (Denmark)

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

    1994-01-01

    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

    OpenAIRE

    Vanderpool, Carin K.; Armstrong, Sandra K.

    2001-01-01

    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.

    OpenAIRE

    Peppler, M S; Schrumpf, M E

    1984-01-01

    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

    Science.gov (United States)

    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

    Science.gov (United States)

    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

    OpenAIRE

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

    2012-01-01

    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

    Directory of Open Access Journals (Sweden)

    Levillain Erwan

    2009-07-01

    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.

    OpenAIRE

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

    1994-01-01

    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.

    OpenAIRE

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

    1996-01-01

    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

    Directory of Open Access Journals (Sweden)

    Walsh, Paul

    2008-08-01

    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.

    Science.gov (United States)

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

    2014-01-01

    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

    Directory of Open Access Journals (Sweden)

    Sandeep Kumar

    2014-01-01

    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

    Science.gov (United States)

    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.

    OpenAIRE

    Ishikawa, H.; Isayama, Y

    1986-01-01

    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

    OpenAIRE

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

    2012-01-01

    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.

    OpenAIRE

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

    1986-01-01

    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

    Science.gov (United States)

    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].

    Science.gov (United States)

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

    2015-09-01

    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

    Directory of Open Access Journals (Sweden)

    Schneiker-Bekel Susanne

    2008-09-01

    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

    OpenAIRE

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

    1999-01-01

    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.

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    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.

    OpenAIRE

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

    1996-01-01

    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.

    LENUS (Irish Health Repository)

    Grogan, Juanita A

    2011-06-01

    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.

    Science.gov (United States)

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

    2012-11-01

    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

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    Valérie Bouchez

    2015-09-01

    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

    Science.gov (United States)

    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?

    Czech Academy of Sciences Publication Activity Database

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

    2014-01-01

    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

    NARCIS (Netherlands)

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

    2014-01-01

    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

    Science.gov (United States)

    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

    NARCIS (Netherlands)

    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.

    2011-01-01

    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

    OpenAIRE

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

    2014-01-01

    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

    NARCIS (Netherlands)

    Hijnen, Marcel

    2006-01-01

    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)

    Science.gov (United States)

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

    1980-01-01

    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

    Science.gov (United States)

    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

    OpenAIRE

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

    1999-01-01

    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.

    Science.gov (United States)

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

    2003-09-01

    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

    OpenAIRE

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

    2011-01-01

    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

    Directory of Open Access Journals (Sweden)

    Patricia Tomé Sandoval

    2008-08-01

    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

    OpenAIRE

    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.

    2014-01-01

    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.

    Science.gov (United States)

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

    2012-12-01

    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

    OpenAIRE

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

    2012-01-01

    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

    OpenAIRE

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

    2013-01-01

    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.

    Directory of Open Access Journals (Sweden)

    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.

    OpenAIRE

    Buck, G E

    1996-01-01

    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

    International Nuclear Information System (INIS)

    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

    OpenAIRE

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

    2013-01-01

    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

    OpenAIRE

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

    2005-01-01

    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

    OpenAIRE

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

    2014-01-01

    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.

    Directory of Open Access Journals (Sweden)

    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.

    Science.gov (United States)

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

    1996-11-01

    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

    OpenAIRE

    Register, Karen B.; Yersin, Andrew G.

    2005-01-01

    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

    Directory of Open Access Journals (Sweden)

    Mohammad Rubayet Hasan

    2014-01-01

    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

    Science.gov (United States)

    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.

    Science.gov (United States)

    Helting, T B; Blackkolb, F

    1981-04-01

    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

    Directory of Open Access Journals (Sweden)

    Park Jihye

    2012-10-01

    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

    Directory of Open Access Journals (Sweden)

    Algecira N.

    1998-12-01

    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.

    Science.gov (United States)

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

    2011-12-01

    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

    Directory of Open Access Journals (Sweden)

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

    2012-10-01

    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.

    Science.gov (United States)

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

    2016-02-01

    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

    OpenAIRE

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

    2005-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2014-01-01

    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.

    OpenAIRE

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

    1989-01-01

    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.

    OpenAIRE

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

    1990-01-01

    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

    Directory of Open Access Journals (Sweden)

    Ossewaarde Jacobus M

    2011-01-01

    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.

    Science.gov (United States)

    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

    2015-04-01

    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

    Directory of Open Access Journals (Sweden)

    Hui Zhang

    2013-10-01

    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.

    Science.gov (United States)

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

    2014-04-01

    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.

    Science.gov (United States)

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

    2000-08-01

    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

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    Chattip Kurehong

    2015-04-01

    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

    Institute of Scientific and Technical Information of China (English)

    JI Quan-an

    2014-01-01

    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.

    Science.gov (United States)

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

    2016-06-01

    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.

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    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.

    OpenAIRE

    Ishikawa, H.; Isayama, Y

    1997-01-01

    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.

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    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.

    OpenAIRE

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

    1986-01-01

    [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

    Czech Academy of Sciences Publication Activity Database

    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.

    2007-01-01

    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

    Science.gov (United States)

    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

    Science.gov (United States)

    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

    Czech Academy of Sciences Publication Activity Database

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

    2015-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2005-01-01

    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

    Institute of Scientific and Technical Information of China (English)

    CHI Shen; SUN Yun; ZHANG Bao-yuan

    2008-01-01

    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.

    Science.gov (United States)

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

    1993-01-25

    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.

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    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.

    Science.gov (United States)

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

    2016-07-01

    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

    Directory of Open Access Journals (Sweden)

    Ernesto Marcos

    2010-01-01

    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

    Directory of Open Access Journals (Sweden)

    Slack Andrew T

    2006-03-01

    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

    Directory of Open Access Journals (Sweden)

    RODRIGO OSSES A

    2010-03-01

    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.

    OpenAIRE

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

    1990-01-01

    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.

    OpenAIRE

    Molina, N C; Parker, C D

    1990-01-01

    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].

    Science.gov (United States)

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

    2016-07-01

    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

    Directory of Open Access Journals (Sweden)

    K. Kamachi

    2015-11-01

    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

    Energy Technology Data Exchange (ETDEWEB)

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

    2014-10-10

    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.

    Science.gov (United States)

    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

    2014-01-01

    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.

    Science.gov (United States)

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

    2016-05-01

    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.

    Directory of Open Access Journals (Sweden)

    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

    Science.gov (United States)

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

    2016-01-01

    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.

    Directory of Open Access Journals (Sweden)

    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

    OpenAIRE

    Pradel, Elizabeth; Locht, Camille

    2001-01-01

    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.

    Directory of Open Access Journals (Sweden)

    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.

    Science.gov (United States)

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

    2016-06-01

    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

    International Nuclear Information System (INIS)

    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.

    Science.gov (United States)

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

    2013-06-01

    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

    Science.gov (United States)

    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

    2016-01-01

    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.

    Science.gov (United States)

    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

    2016-07-01

    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

    Czech Academy of Sciences Publication Activity Database

    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.

    Science.gov (United States)

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

    1992-12-01

    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.

    Science.gov (United States)

    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

    2002-06-21

    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.

    Science.gov (United States)

    Monack, D M; Falkow, S

    1993-11-01

    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.

    Science.gov (United States)

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

    2015-08-01

    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

    International Nuclear Information System (INIS)

    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.

    Directory of Open Access Journals (Sweden)

    Ladislav Bumba

    2010-05-01

    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

    Directory of Open Access Journals (Sweden)

    Yilian Plasencia,

    2000-11-01

    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.

    Science.gov (United States)

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

    2016-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2013-01-01

    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

    OpenAIRE

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

    2015-01-01

    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.

    Science.gov (United States)

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

    2016-03-01

    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

    Directory of Open Access Journals (Sweden)

    Sebastião Timo Iaria

    1977-09-01

    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

    Directory of Open Access Journals (Sweden)

    Tania Alen Coutinho

    2009-09-01

    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.

    Directory of Open Access Journals (Sweden)

    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

    Institute of Scientific and Technical Information of China (English)

    赵宁; 王芳; 恽时锋; 范志宇; 胡波

    2011-01-01

    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

    Czech Academy of Sciences Publication Activity Database

    Vojtová, Jana; Kamanová, Jana; Šebo, Peter

    2006-01-01

    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

  5. [PERSISTENCE OF BORDETELLA PERTUSSIS BACTERIA AND A POSSIBLE MECHANISM OF ITS FORMATION].

    Science.gov (United States)

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

    2015-01-01

    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

    Science.gov (United States)

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

    1993-07-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2009-01-01

    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.

    Science.gov (United States)

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

    2016-06-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2013-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2014-01-01

    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].

    Science.gov (United States)

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

    1985-09-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2003-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2015-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2010-01-01

    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

    NARCIS (Netherlands)

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

    2014-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2015-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2001-01-01

    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

    OpenAIRE

    Gaillard, María Emilia

    2009-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2006-01-01

    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

    Czech Academy of Sciences Publication Activity Database

    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.

    2005-01-01

    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

    Directory of Open Access Journals (Sweden)

    Valladares-Carranza B.

    2011-10-01

    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

    OpenAIRE

    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.

    2011-01-01

    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

    OpenAIRE

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

    2015-01-01

    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

    International Nuclear Information System (INIS)

    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

    Czech Academy of Sciences Publication Activity Database

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

    2015-01-01

    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+

    Czech Academy of Sciences Publication Activity Database

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

    2001-01-01

    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

    Science.gov (United States)

    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

    Czech Academy of Sciences Publication Activity Database

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

    2004-01-01

    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

    International Nuclear Information System (INIS)

    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

    OpenAIRE

    Asokanathan, Catpagavalli; Corbel, Michael; Xing, Dorothy

    2013-01-01

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Czech Academy of Sciences Publication Activity Database

    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

    2014-01-01

    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.

    Directory of Open Access Journals (Sweden)

    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

    OpenAIRE

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

    2013-01-01

    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

    Czech Academy of Sciences Publication Activity Database

    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

    Czech Academy of Sciences Publication Activity Database

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

    2008-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2015-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2010-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2006-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2015-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2012-01-01

    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?

    Science.gov (United States)

    2015-01-01

    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

    Science.gov (United States)

    2013-04-03

    ..., 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

    Lifescience Database Archive (English)

    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

    Science.gov (United States)

    2012-12-31

    ... 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

    Czech Academy of Sciences Publication Activity Database

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

    2006-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2013-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2000-01-01

    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

    Lifescience Database Archive (English)

    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.

    Science.gov (United States)

    2010-04-01

    ... 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

    Science.gov (United States)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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 t...id 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

    Lifescience Database Archive (English)

    Full Text Available :none) Ralstonia eutropha JMP134 chrom... 97 5e-19 AP006841_3502( AP006841 |pid:none) Bacteroides fragi...om... 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 stra...in 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

    Institute of Scientific and Technical Information of China (English)

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

    2015-01-01

    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

    Institute of Scientific and Technical Information of China (English)

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

    2015-01-01

    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

    Lifescience Database Archive (English)

    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=3.1.13.1; ... 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

    Directory of Open Access Journals (Sweden)

    Arbefeville S

    2015-09-01

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    NARCIS (Netherlands)

    Schure, R.M.

    2014-01-01

    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

    Index Scriptorium Estoniae

    Oona, Marje, 1963-

    2012-01-01

    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

    OpenAIRE

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

    2008-01-01

    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.

    Science.gov (United States)

    2010-04-01

    .... 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

    DEFF Research Database (Denmark)

    Claesson, Mogens Helweg

    2011-01-01

    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

    OpenAIRE

    MacLean, D. W.

    1982-01-01

    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.

    Science.gov (United States)

    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

    2014-03-01

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    OpenAIRE

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

    2000-01-01

    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

    OpenAIRE

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

    2012-01-01

    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

    Directory of Open Access Journals (Sweden)

    Workalemahu M. Berhanu

    2012-07-01

    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

    OpenAIRE

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

    1997-01-01

    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†

    OpenAIRE

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

    2009-01-01

    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

    Science.gov (United States)

    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

    2012-01-01

    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

    Czech Academy of Sciences Publication Activity Database

    Šebo, Peter; Osička, Radim; Mašín, Jiří

    2014-01-01

    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

    Czech Academy of Sciences Publication Activity Database

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

    2013-01-01

    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

    OpenAIRE

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

    2010-01-01

    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.

    Directory of Open Access Journals (Sweden)

    Tanizaki,Yoshiro

    1983-06-01

    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.

    OpenAIRE

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

    1983-01-01

    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.

    Science.gov (United States)

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

    2015-10-01

    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

    Directory of Open Access Journals (Sweden)

    Celso Pérez-Bolaños

    2011-01-01

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    DEFF Research Database (Denmark)

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

    2007-01-01

    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

    OpenAIRE

    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

    2014-01-01

    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.

    Science.gov (United States)

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

    2016-01-01

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    OpenAIRE

    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

    2008-01-01

    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.

    Directory of Open Access Journals (Sweden)

    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

    Directory of Open Access Journals (Sweden)

    Vicente Amato Neto

    1977-08-01

    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.

    Directory of Open Access Journals (Sweden)

    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.

    Science.gov (United States)

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

    2010-01-01

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    Full Text Available 7 G02257( G02257 ) NAD(P) transhydrogenase (B-specific) (EC 1.6.1.1)... 424 e-117 AM920436_1880( AM920436 |p...... 417 e-114 S54876( S54876 ) NAD(P) transhydrogenase (B-specific) (EC 1.6.1.1)... 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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    Full Text Available total letters Score E Sequences producing significant alignments: (bits) Value Contig-U14087-1 (Contig-U14087-1Q) /CSM_Contig/Conti...ne S-transferase Y-b subu... 55 2e-06 (Q9JHF7) RecName: Full=Glutathione-requiring prostaglandin D s...id:none) 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.

    Science.gov (United States)

    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

    2001-05-01

    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.

    Science.gov (United States)

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

    2014-01-01

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Energy Technology Data Exchange (ETDEWEB)

    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)

    2010-10-15

    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

    Lifescience Database Archive (English)

    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( ...ve DEAD box helicase p... 50 0.22 1 ( AY070052 ) Arabidopsis thaliana putati

  14. Dicty_cDB: Contig-U12487-1 [Dicty_cDB

    Lifescience Database Archive (English)

    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) Mic...us 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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Science.gov (United States)

    Hejtmancik, Kelly E.

    1987-01-01

    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.

    Science.gov (United States)

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

    2015-03-01

    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.

    Science.gov (United States)

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

    2015-09-01

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    Lifescience Database Archive (English)

    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

    DEFF Research Database (Denmark)

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

    2003-01-01

    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

    Lifescience Database Archive (English)

    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 discoide...um 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

    Directory of Open Access Journals (Sweden)

    Vicente Amato Neto

    1977-08-01

    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

    Lifescience Database Archive (English)

    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

    Directory of Open Access Journals (Sweden)

    Hiromi eSato

    2011-07-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2010-09-08

    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.

  11. Diverse nucleotide compositions and sequence fluctuation in Rubisco protein genes

    Science.gov (United States)

    Holden, Todd; Dehipawala, S.; Cheung, E.; Bienaime, R.; Ye, J.; Tremberger, G., Jr.; Schneider, P.; Lieberman, D.; Cheung, T.

    2011-10-01

    The Rubisco protein-enzyme is arguably the most abundance protein on Earth. The biology dogma of transcription and translation necessitates the study of the Rubisco genes and Rubisco-like genes in various species. Stronger correlation of fractal dimension of the atomic number fluctuation along a DNA sequence with Shannon entropy has been observed in the studied Rubisco-like gene sequences, suggesting a more diverse evolutionary pressure and constraints in the Rubisco sequences. The strategy of using metal for structural stabilization appears to be an ancient mechanism, with data from the porphobilinogen deaminase gene in Capsaspora owczarzaki and Monosiga brevicollis. Using the chi-square distance probability, our analysis supports the conjecture that the more ancient Rubisco-like sequence in Microcystis aeruginosa would have experienced very different evolutionary pressure and bio-chemical constraint as compared to Bordetella bronchiseptica, the two microbes occupying either end of the correlation graph. Our exploratory study would indicate that high fractal dimension Rubisco sequence would support high carbon dioxide rate via the Michaelis- Menten coefficient; with implication for the control of the whooping cough pathogen Bordetella bronchiseptica, a microbe containing a high fractal dimension Rubisco-like sequence (2.07). Using the internal comparison of chi-square distance probability for 16S rRNA (~ E-22) versus radiation repair Rec-A gene (~ E-05) in high GC content Deinococcus radiodurans, our analysis supports the conjecture that high GC content microbes containing Rubisco-like sequence are likely to include an extra-terrestrial origin, relative to Deinococcus radiodurans. Similar photosynthesis process that could utilize host star radiation would not compete with radiation resistant process from the biology dogma perspective in environments such as Mars and exoplanets.

  12. Antimicrobial susceptibility monitoring of bacterial pathogens isolated from respiratory tract infections in dogs and cats across Europe: ComPath results.

    Science.gov (United States)

    Morrissey, Ian; Moyaert, Hilde; de Jong, Anno; El Garch, Farid; Klein, Ulrich; Ludwig, Carolin; Thiry, Julien; Youala, Myriam

    2016-08-15

    ComPath is a pan-European resistance monitoring programme collecting bacterial pathogens from dogs and cats. We present data for respiratory tract infection (RTI) isolates collected between 2008 and 2010. Antimicrobial minimal inhibitory concentrations (MICs) were determined and susceptibility calculated following Clinical Laboratory Standards Institute (CLSI) standards for veterinary medicine. The main pathogen from dogs was Staphylococcus intermedius Group (49/215, 22.8%) which was >90% susceptible to most antimicrobials (including oxacillin - 93.9%; 3 isolates confirmed mecA-positive) but only 59.2%, 73.5% and 87.8% susceptible to tetracycline, chloramphenicol and penicillin. Bordetella bronchiseptica (48/215, 22.3%), streptococci (36/215, 16.7%), Escherichia coli (24/215, 11.2%) and Pasteurella multocida (23/215, 10.7%) were also found in dog RTI. There are no breakpoints for Bordetella bronchiseptica. Most streptococci were penicillin- chloramphenicol-, ampicillin- and pradofloxacin-susceptible. None were enrofloxacin-resistant but 6 isolates (16.7%) were of intermediate susceptibility. The least active agent against streptococci was tetracycline (47.2% susceptible). For E. coli, 37.5% were ampicillin-susceptible but 83.3% were amoxicillin/clavulanic acid-susceptible. Only chloramphenicol showed susceptibility>90% against E. coli, with 66.7% tetracycline-susceptible and 79.2% to 87.5% susceptibility to enrofloxacin, trimethoprim-sulfamethoxazole or pradofloxacin. P. multocida were susceptible to pradofloxacin (no other breakpoints are available). The main pathogen from cats was P. multocida (82/186, 44.1%), where only pradofloxacin has breakpoints (100% susceptible). Streptococci were also collected from cats (25/186, 13.4%) and were >90% susceptible to all antimicrobials except tetracycline (36% susceptible). Most susceptibility was calculated with human-derived breakpoints and some antimicrobials had no breakpoints. Therefore predictions of clinical utility

  13. Use of immunomodulators in infectious diseases of domestic animals/ Uso de imunomoduladores nas enfermidades infecciosas dos animais domésticos

    Directory of Open Access Journals (Sweden)

    Jane Megid

    2007-08-01

    Full Text Available Immunomodulators are substances that act in the immune system providing, increase of the organic answer against microorganisms, including virus, bacteria and protozoa, by inducing the production of interferon and its inducers. There are a lot of situations in veterinary medicine where it is usefull to potencialize the immune response of individuals, mainly when is desired to increase the resistance to infections and the treatment of immunossupressing or multifactorials infectious diseases. In veterinary medicine some of more used immunomodulators are interferons and interferon inducers, interleukines, Baccilus of Calmett-Guérin (BCG and its derivated, Propionibacterium acnes (Corynebacterium parvum, mixed bacterial vaccine, PIND-ORF, Phosprenyl, Quillja Saponis, Bordetella pertussis, avridine and the levamizole. The present work review the available scientific literature, regarding the use of different immunomodulators in the prophylaxis and in the therapeutics of infectious diseases in domestic animals.Imunomoduladores são substâncias que atuam no sistema imunológico conferindo aumento da resposta orgânica contra determinados microorganismos, incluindo vírus, bactérias e protozoários, mediante à produção de interferon e seus indutores. Existem muitas situações na medicina veterinária em que se torna desejável potencializar a resposta imune, principalmente quando se pretende aumentar a resistência às infecções e no tratamento de enfermidades imunossupressoras ou de doenças infecciosas multifatorias, ou seja, nas quais vários agentes estão envolvidos e devido a isso, dificilmente obtêm-se sucesso no emprego de tratamentos convencionais. Na medicina veterinária alguns imunomoduladores utilizados são interferons , interleucinas, Bacilo de Calmett-Guérin (BCG e seus derivados, Propionibacterium acnes (Corynebacterium parvum, vacina bacteriana mista, PIND-ORF, Phosprenyl, Quillaja saponis, Bordetella pertussis, avridina e

  14. Brucella abortus ornithine lipids are dispensable outer membrane components devoid of a marked pathogen-associated molecular pattern.

    Directory of Open Access Journals (Sweden)

    Leyre Palacios-Chaves

    Full Text Available The brucellae are α-Proteobacteria facultative intracellular parasites that cause an important zoonosis. These bacteria escape early detection by innate immunity, an ability associated to the absence of marked pathogen-associated molecular patterns in the cell envelope lipopolysaccharide, lipoproteins and flagellin. We show here that, in contrast to the outer membrane ornithine lipids (OL of other Gram negative bacteria, Brucella abortus OL lack a marked pathogen-associated molecular pattern activity. We identified two OL genes (olsB and olsA and by generating the corresponding mutants found that olsB deficient B. abortus did not synthesize OL or their lyso-OL precursors. Liposomes constructed with B. abortus OL did not trigger IL-6 or TNF-α release by macrophages whereas those constructed with Bordetella pertussis OL and the olsB mutant lipids as carriers were highly active. The OL deficiency in the olsB mutant did not promote proinflammatory responses or generated attenuation in mice. In addition, OL deficiency did not increase sensitivity to polymyxins, normal serum or complement consumption, or alter the permeability to antibiotics and dyes. Taken together, these observations indicate that OL have become dispensable in the extant brucellae and are consistent within the trend observed in α-Proteobacteria animal pathogens to reduce and eventually eliminate the envelope components susceptible of recognition by innate immunity.

  15. Formaldehyde-inactivated human enterovirus 71 vaccine is compatible for co-immunization with a commercial pentavalent vaccine.

    Science.gov (United States)

    Chen, Chun-Wei; Lee, Yi-Ping; Wang, Ya-Fang; Yu, Chun-Keung

    2011-03-24

    In this study we tested the effectiveness of a formaldehyde-inactivated EV71 vaccine and its compatibility for co-immunization with a pentavalent vaccine that contained inactivated poliovirus (PV) vaccine. The inactivated EV71 vaccine (C2 genogroup) elicited an antibody response which broadly neutralized homologous and heterologous genogroups, including B4, C4, and B5. Pups from vaccinated dams were resistant to the EV71 challenge and had a high survival rate and a low tissue viral burden when compared to those from non-vaccinated counterparts. Co-immunization with pentavalent and inactivated EV71 vaccines elicited antibodies against the major components of the pentavalent vaccine including the PV, Bordetella pertussis, Haemophilus influenzae type b, diphtheria toxoid, and tetanus toxoid at the same levels as in mice immunized with pentavalent vaccine alone. Likewise, EV71 neutralizing antibody titers were comparable between EV71-vaccinated mice and mice co-immunized with the two vaccines. These results indicate that formaldehyde-inactivated whole virus EV71 vaccine is feasible for designing multivalent vaccines. PMID:21315698

  16. Molecular surveillance of traditional and emerging pathogens associated with canine infectious respiratory disease.

    Science.gov (United States)

    Decaro, Nicola; Mari, Viviana; Larocca, Vittorio; Losurdo, Michele; Lanave, Gianvito; Lucente, Maria Stella; Corrente, Marialaura; Catella, Cristiana; Bo, Stefano; Elia, Gabriella; Torre, Giorgio; Grandolfo, Erika; Martella, Vito; Buonavoglia, Canio

    2016-08-30

    A molecular survey for traditional and emerging pathogens associated with canine infectious respiratory disease (CIRD) was conducted in Italy between 2011 and 2013 on a total of 138 dogs, including 78 early acute clinically ill CIRD animals, 22 non-clinical but exposed to clinically ill CIRD dogs and 38 CIRD convalescent dogs. The results showed that canine parainfluenza virus (CPIV) was the most commonly detected CIRD pathogen, followed by canine respiratory coronavirus (CRCoV), Bordetella bronchiseptica, Mycoplasma cynos, Mycoplasma canis and canine pneumovirus (CnPnV). Some classical CIRD agents, such as canine adenoviruses, canine distemper virus and canid herpesvirus 1, were not detected at all, as were not other emerging respiratory viruses (canine influenza virus, canine hepacivirus) and bacteria (Streptococcus equi subsp. zooepidemicus). Most severe forms of respiratory disease were observed in the presence of CPIV, CRCoV and M. cynos alone or in combination with other pathogens, whereas single CnPnV or M. canis infections were detected in dogs with no or very mild respiratory signs. Interestingly, only the association of M. cynos (alone or in combination with either CRCoV or M. canis) with severe clinical forms was statistically significant. The study, while confirming CPIV as the main responsible for CIRD occurrence, highlights the increasing role of recently discovered viruses, such as CRCoV and CnPnV, for which effective vaccines are not available in the market. PMID:27527760

  17. Age-related presence of selected viral and bacterial pathogens in paraffin-embedded lung samples of dogs with pneumonia.

    Science.gov (United States)

    Wöhrer, Daniela; Spergser, Joachim; Bagrinovschi, Gabriela; Möstl, Karin; Weissenböck, Herbert

    2016-03-01

    The aim of this retrospective study was to detect selected pathogens in pneumonic lung tissue of dogs of different age groups by immunohistochemistry (IHC), in situ hybridisation (ISH) or polymerase chain reaction (PCR) in order to get information about their involvement in pneumonia formation. In archived formalin-fixed and paraffin wax-embedded lung samples from 68 cases with the clinical and histologic diagnosis of pneumonia the histological pattern of pneumonia was re-evaluated and the samples were further investigated for the following infectious agents: canine distemper virus (CDV), canine adenovirus type 2 (CAV-2), canine respiratory coronavirus (CRCoV), Bordetella (B.) bronchiseptica, Pasteurella (P.) multocida, Mycoplasma spp., and Pneumocystis spp. In 47.1% of the samples at least one of the featured respiratory pathogens was detected. In 31.3% of these positive samples more than one pathogen could be found. The correct detection of CDV had been achieved in ten out of eleven positive cases (90.9%) upon initial investigation, but the presence of bacterial pathogens, like B. bronchiseptica (10 cases) and P. multocida (17 cases) had been missed in all but one case. While CDV and CRCoV infections were exclusively found in dogs younger than one year, the vast majority of infections with P. multocida and B. bronchiseptica were both common either in dogs younger than 4 months or older than one year. Thus, this retrospective approach yielded valuable data on the presence, absence and prevalence of certain respiratory pathogens in dogs with pneumonia. PMID:26919147

  18. Serum antibody responses to vaccinal antigens in lean and obese geriatric dogs.

    Science.gov (United States)

    Ellis, John; Gow, Sheryl; Rhodes, Carrie; Lacoste, Stacey; Kong, Lyndsay; Musil, Kristyna; Snead, Elisabeth

    2016-05-01

    The immune responses in control dogs [1 to 4 years of age, body condition score (BCS): 4 to 5 out of 9] were compared to those of aging dogs (based on breed and body size) either categorized as lean (BCS: 4 to 5 out of 9) or obese (BCS: 8 to 9 out of 9). Of interest were the serum titers to the following common agents found in vaccines, canine parainfluenza virus (CPIV), canine parvovirus (CPV), canine distemper virus (CDV), canine respiratory coronavirus (CRCoV), and Bordetella bronchiseptica. There were no statistical differences in the antibodies to CPIV, B. bronchispetica, and CRCoV, among the age/weight categories, nor among the age/weight categories and the time, in days, between the date of sample collection and the date of the last recorded vaccination for CPIV, B. bronchiseptica, CPV, and CDV. For CPV, the control dogs had significantly (P < 0.002) higher serum neutralization (SN) titers than the lean geriatric dogs and the obese geriatric dogs. For CDV SN titers, the only statistically significant (P = 0.01) difference was that the control dogs had higher SN titers than the lean geriatric dogs. PMID:27152043

  19. Comparison of point-of-care-compatible lysis methods for bacteria and viruses.

    Science.gov (United States)

    Heiniger, Erin K; Buser, Joshua R; Mireles, Lillian; Zhang, Xiaohong; Ladd, Paula D; Lutz, Barry R; Yager, Paul

    2016-09-01

    Nucleic acid sample preparation has been an especially challenging barrier to point-of-care nucleic acid amplification tests in low-resource settings. Here we provide a head-to-head comparison of methods for lysis of, and nucleic acid release from, several pathogenic bacteria and viruses-methods that are adaptable to point-of-care usage in low-resource settings. Digestion with achromopeptidase, a mixture of proteases and peptidoglycan-specific hydrolases, followed by thermal deactivation in a boiling water bath, effectively released amplifiable nucleic acid from Staphylococcus aureus, Bordetella pertussis, respiratory syncytial virus, and influenza virus. Achromopeptidase was functional after dehydration and reconstitution, even after eleven months of dry storage without refrigeration. Mechanical lysis methods proved to be effective against a hard-to-lyse Mycobacterium species, and a miniature bead-mill, the AudioLyse, is shown to be capable of releasing amplifiable DNA and RNA from this species. We conclude that point-of-care-compatible sample preparation methods for nucleic acid tests need not introduce amplification inhibitors, and can provide amplification-ready lysates from a wide range of bacterial and viral pathogens. PMID:27424294

  20. Polyethersulfone improves isothermal nucleic acid amplification compared to current paper-based diagnostics.

    Science.gov (United States)

    Linnes, J C; Rodriguez, N M; Liu, L; Klapperich, C M

    2016-04-01

    Devices based on rapid, paper-based, isothermal nucleic acid amplification techniques have recently emerged with the potential to fill a growing need for highly sensitive point-of-care diagnostics throughout the world. As this field develops, such devices will require optimized materials that promote amplification and sample preparation. Herein, we systematically investigated isothermal nucleic acid amplification in materials currently used in rapid diagnostics (cellulose paper, glass fiber, and nitrocellulose) and two additional porous membranes with upstream sample preparation capabilities (polyethersulfone and polycarbonate). We compared amplification efficiency from four separate DNA and RNA targets (Bordetella pertussis, Chlamydia trachomatis, Neisseria gonorrhoeae, and Influenza A H1N1) within these materials using two different isothermal amplification schemes, helicase dependent amplification (tHDA) and loop-mediated isothermal amplification (LAMP), and traditional PCR. We found that the current paper-based diagnostic membranes inhibited nucleic acid amplification when compared to membrane-free controls; however, polyethersulfone allowed for efficient amplification in both LAMP and tHDA reactions. Further, observing the performance of traditional PCR amplification within these membranes was not predicative of their effects on in situ LAMP and tHDA. Polyethersulfone is a new material for paper-based nucleic acid amplification, yet provides an optimal support for rapid molecular diagnostics for point-of-care applications. PMID:26906904

  1. Sequential unfolding of beta helical protein by single-molecule atomic force microscopy.

    Directory of Open Access Journals (Sweden)

    David Alsteens

    Full Text Available The parallel βhelix is a common fold among extracellular proteins, however its mechanical properties remain unexplored. In Gram-negative bacteria, extracellular proteins of diverse functions of the large 'TpsA' family all fold into long βhelices. Here, single-molecule atomic force microscopy and steered molecular dynamics simulations were combined to investigate the mechanical properties of a prototypic TpsA protein, FHA, the major adhesin of Bordetella pertussis. Strong extension forces were required to fully unfold this highly repetitive protein, and unfolding occurred along a stepwise, hierarchical process. Our analyses showed that the extremities of the βhelix unfold early, while central regions of the helix are more resistant to mechanical unfolding. In particular, a mechanically resistant subdomain conserved among TpsA proteins and critical for secretion was identified. This nucleus harbors structural elements packed against the βhelix that might contribute to stabilizing the N-terminal region of FHA. Hierarchical unfolding of the βhelix in response to a mechanical stress may maintain β-helical portions that can serve as templates for regaining the native structure after stress. The mechanical properties uncovered here might apply to many proteins with β-helical or related folds, both in prokaryotes and in eukaryotes, and play key roles in their structural integrity and functions.

  2. Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling.

    Science.gov (United States)

    Barbier, Mariette; Damron, F Heath

    2016-01-01

    Since their discovery, fluorescent proteins have been widely used to study protein function, localization or interaction, promoter activity and regulation, drug discovery or for non-invasive imaging. They have been extensively modified to improve brightness, stability, and oligomerization state. However, only a few studies have focused on understanding the dynamics of fluorescent proteins expression in bacteria. In this work, we developed a set plasmids encoding 12 fluorescent proteins for bacterial labeling to facilitate the study of pathogen-host interactions. These broad-spectrum plasmids can be used with a wide variety of Gram-negative microorganisms including Escherichia coli, Pseudomonas aeruginosa, Burkholderia cepacia, Bordetella bronchiseptica, Shigella flexneri or Klebsiella pneumoniae. For comparison, fluorescent protein expression and physical characteristics in Escherichia coli were analyzed using fluorescence microscopy, flow cytometry and in vivo imaging. Fluorescent proteins derived from the Aequorea Victoria family showed high photobleaching, while proteins form the Discosoma sp. and the Fungia coccina family were more photostable for microscopy applications. Only E2-Crimson, mCherry and mKeima were successfully detected for in vivo applications. Overall, E2-Crimson was the fastest maturing protein tested in E. coli with the best overall performance in the study parameters. This study provides a unified comparison and comprehensive characterization of fluorescent protein photostability, maturation and toxicity, and offers general recommendations on the optimal fluorescent proteins for in vitro and in vivo applications. PMID:26937640

  3. Bacterial outer membrane vesicles and vaccine applications.

    Science.gov (United States)

    Acevedo, Reinaldo; Fernández, Sonsire; Zayas, Caridad; Acosta, Armando; Sarmiento, Maria Elena; Ferro, Valerie A; Rosenqvist, Einar; Campa, Concepcion; Cardoso, Daniel; Garcia, Luis; Perez, Jose Luis

    2014-01-01

    Vaccines based on outer membrane vesicles (OMV) were developed more than 20 years ago against Neisseria meningitidis serogroup B. These nano-sized structures exhibit remarkable potential for immunomodulation of immune responses and delivery of meningococcal antigens or unrelated antigens incorporated into the vesicle structure. This paper reviews different applications in OMV Research and Development (R&D) and provides examples of OMV developed and evaluated at the Finlay Institute in Cuba. A Good Manufacturing Practice (GMP) process was developed at the Finlay Institute to produce OMV from N. meningitidis serogroup B (dOMVB) using detergent extraction. Subsequently, OMV from N. meningitidis, serogroup A (dOMVA), serogroup W (dOMVW), and serogroup X (dOMVX) were obtained using this process. More recently, the extraction process has also been applied effectively for obtaining OMV on a research scale from Vibrio cholerae (dOMVC), Bordetella pertussis (dOMVBP), Mycobacterium smegmatis (dOMVSM), and BCG (dOMVBCG). The immunogenicity of the OMV has been evaluated for specific antibody induction, and together with functional bactericidal and challenge assays in mice has shown their protective potential. dOMVB has been evaluated with non-neisserial antigens, including with a herpes virus type 2 glycoprotein, ovalbumin, and allergens. In conclusion, OMV are proving to be more versatile than first conceived and remain an important technology for development of vaccine candidates. PMID:24715891

  4. BACTERIAL OUTER MEMBRANE VESICLES AND VACCINE APPLICATIONS

    Directory of Open Access Journals (Sweden)

    Reinaldo eAcevedo

    2014-03-01

    Full Text Available Vaccines based on outer membrane vesicles (OMV were developed more than 20 years ago against Neisseria meningitidis serogroup B. These nano-sized structures exhibit remarkable potential for immunomodulation of immune responses and delivery of self meningococcal antigens or unrelated antigens incorporated into the vesicle structure. This paper reviews different applications in OMV Research and Development (R&D and provides examples of OMV developed and evaluated at the Finlay Institute in Cuba. A Good Manufacturing Practice (GMP process was developed at the Finlay Institute to produce OMV from N. meningitidis serogroup B (dOMVB using detergent extraction. Subsequently, OMV from N. meningitidis, serogroup A (dOMVA, serogroup W (dOMVW and serogroup X (dOMVX were obtained using this process. More recently, the extraction process has also been applied effectively for obtaining OMV on a research scale from Vibrio cholerae (dOMVC, Bordetella pertussis (dOMVBP, Mycobacterium smegmatis (dOMVSM and BCG (dOMVBCG. The immunogenicity of the OMV have been evaluated for specific antibody induction, and together with functional bactericidal and challenge assays in mice have shown their protective potential. dOMVB has been evaluated with non-self neisserial antigens, including with a herpes virus type 2 glycoprotein, ovalbumin and allergens. In conclusion, OMV are proving to be more versatile than first conceived and remain an important technology for development of vaccine candidates.

  5. Pertussis: Microbiology, Disease, Treatment, and Prevention.

    Science.gov (United States)

    Kilgore, Paul E; Salim, Abdulbaset M; Zervos, Marcus J; Schmitt, Heinz-Josef

    2016-07-01

    Pertussis is a severe respiratory infection caused by Bordetella pertussis, and in 2008, pertussis was associated with an estimated 16 million cases and 195,000 deaths globally. Sizeable outbreaks of pertussis have been reported over the past 5 years, and disease reemergence has been the focus of international attention to develop a deeper understanding of pathogen virulence and genetic evolution of B. pertussis strains. During the past 20 years, the scientific community has recognized pertussis among adults as well as infants and children. Increased recognition that older children and adolescents are at risk for disease and may transmit B. pertussis to younger siblings has underscored the need to better understand the role of innate, humoral, and cell-mediated immunity, including the role of waning immunity. Although recognition of adult pertussis has increased in tandem with a better understanding of B. pertussis pathogenesis, pertussis in neonates and adults can manifest with atypical clinical presentations. Such disease patterns make pertussis recognition difficult and lead to delays in treatment. Ongoing research using newer tools for molecular analysis holds promise for improved understanding of pertussis epidemiology, bacterial pathogenesis, bioinformatics, and immunology. Together, these advances provide a foundation for the development of new-generation diagnostics, therapeutics, and vaccines. PMID:27029594

  6. Isolation, molecular and biochemical characterization of oil degrading bacteria from contaminated soil at an oil refinery

    International Nuclear Information System (INIS)

    Biodegradation using microorganisms is considered to be cost-effective and environmentally friendly treatment of oil-contaminated sites. Oil-biodegrading bacterial strains were isolated, identified and characterized from oil contaminated soil samples at oil refinery in Zarqa (Jordan). Thirty four bacterial isolates were grown on mineral salt media supplemented with crude oil, but 16 showed positive biodegradation of diesel. All the 34 bacterial isolates were characterized at the molecular and bio-chemical levels, and showed positive polymerase chain reaction (PCR) amplification product size of 1500 bp when 16s rDNA bacterial universal primers were used. Eighteen bacterial isolates showed positive PCR amplification product size of 150 bp specific for the genus Pseudomonas and 3 bacterial isolates showed positive amplification product size of 1500 bp specific for the genus Acinetobacter. Biochemical and physiological characterization performed on the 34 bacterial isolates revealed the presence of oil biodegrading bacterial genera and species of Pseudomonas Acidovorans, P. aeruginosa, P. vesicularis, Acinetobacter calcoaceticus, Ac. lowffii, Micro-ococcus luteus, M. varians, M. lylae, M. roseus, Alcaligenes denitrificians, Bacillus megaterium, Comamonas sp., Moralxella sp., Bordetella sp., P. putida, P. stutzeri and P. mallei. (au)

  7. Nucleotidyl cyclase activity of particulate guanylyl cyclase A: comparison with particulate guanylyl cyclases E and F, soluble guanylyl cyclase and bacterial adenylyl cyclases CyaA and edema factor.

    Directory of Open Access Journals (Sweden)

    Kerstin Y Beste

    Full Text Available Guanylyl cyclases (GCs regulate many physiological processes by catalyzing the synthesis of the second messenger cGMP. The GC family consists of seven particulate GCs (pGCs and a nitric oxide-activated soluble GC (sGC. Rat sGC α1β1 possesses much broader substrate specificity than previously assumed. Moreover, the exotoxins CyaA from Bordetella pertussis and edema factor (EF from Bacillus anthracis possess nucleotidyl cyclase (NC activity. pGC-A is a natriuretic peptide-activated homodimer with two catalytic sites that act cooperatively. Here, we studied the NC activity of rat pGC-A in membranes of stably transfected HEK293 cells using a highly sensitive and specific HPLC-MS/MS technique. GTP and ITP were effective, and ATP and XTP were only poor, pGC-A substrates. In contrast to sGC, pGC-A did not use CTP and UTP as substrates. pGC-E and pGC-F expressed in bovine rod outer segment membranes used only GTP as substrate. In intact HEK293 cells, pGC-A generated only cGMP. In contrast to pGCs, EF and CyaA showed very broad substrate-specificity. In conclusion, NCs exhibit different substrate-specificities, arguing against substrate-leakiness of enzymes and pointing to distinct physiological functions of cyclic purine and pyrimidine nucleotides.

  8. Discovery of Novel Secreted Virulence Factors from Salmonella enterica Serovar Typhimurium by Proteomic Analysis of Culture Supernatants

    Energy Technology Data Exchange (ETDEWEB)

    Niemann, George; Brown, Roslyn N.; Gustin, Jean K.; Stufkens, Afke; Shaikh-Kidwai, Afshan S.; Li, Jie; McDermott, Jason E.; Brewer, Heather M.; Schepmoes, Athena A.; Smith, Richard D.; Adkins, Joshua N.; Heffron, Fred

    2011-01-01

    The intracellular pathogen Salmonella enterica serovar Typhimurium is a leading cause of acute gastroenteritis in the world. This pathogen has two type-III secretion systems (TTSS) necessary for virulence that are encoded in Salmonella pathogenicity islands 1 and 2 (SPI-1 and SPI-2) and are expressed during extracellular or intracellular infectious states, respectively, to deliver virulence factors (effectors) to the host cell cytoplasm. While many have been identified and at least partially characterized, the full repertoire of effectors has not been catalogued. In this mass spectrometry-based proteomics study, we identified effector proteins secreted under minimal acidic medium growth conditions that induced the SPI-2 TTSS and its effectors, and compared the secretome from the parent strain to the secretome from strains missing either essential (SsaK) or regulatory components (SsaL) of the SPI-2 secretion apparatus. We identified 75% of the known TTSS effector repertoire. Excluding translocon components, 95% of the known effectors were biased for identification in the ssaL mutant background, which demonstrated that SsaL regulates SPI-2 type III secretion. To confirm secretion to animal cells, we made translational fusions of several of the best candidates to the calmodulin-dependent adenylate cyclase of Bordetella pertussis and assayed cAMP levels of infected J774 macrophage-like cells. From these infected cells we identified six new TTSS effectors and two others that are secreted independent of TTSS. Our results substantiate reports of additional secretion systems encoded by Salmonella other than TTSS.

  9. Swine infectious agents in Tayassu pecari and Pecari tajacu tissue samples from Brazil.

    Science.gov (United States)

    de Castro, Alessandra Marnie Martins Gomes; Brombila, Talita; Bersano, Josete Garcia; Soares, Herbert Sousa; Silva, Sheila Oliveira de Souza; Minervino, Antonio Humberto Hamad; Ogata, Renato Akio; Gennari, Solange Maria; Richtzenhain, Leonardo Jose

    2014-04-01

    Peccaries and pigs, Tayassuidae and Suidae respectively, diverged approximately one million years ago from a common ancestor. Because these families share some pathogens, peccaries can act as reservoirs of infectious pathogens for domestic and wild swine. We evaluated the presence of swine infectious agents in the spleen and lung tissues of white-lipped peccaries (WLP; Tayassu pecari) and collared peccaries (CP; Pecari tajacu) in Brazil. Samples from 10 adult CP and three WLP, which had been hunted by locals or hit by motor vehicles, were obtained from two free-ranging Brazilian populations. The samples were tested by PCR for Mycoplasma hyopneumoniae, Bordetella bronchiseptica, Pasteurella multocida, porcine circovirus 2 (PCV2), Suid herpesvirus 1 (SuHV-1), and porcine parvovirus (PPV). Positive samples were sequenced. Both species were negative for PPV and B. bronchiseptica and positive for PCV2 and SuHV-1. The lungs of two animals were positive for M. hyopneumoniae and P. multocida. This report is the first demonstration of PCV2 and SuHV-1 swine viruses and of M. hyopneumoniae and P. multocida bacteria in peccaries. One factor contributing to this detection was access to tissue samples, which is uncommon. The role of these infectious agents in peccaries is unknown and further epidemiologic studies should be performed. This study identified several infectious agents in peccaries and highlighted the importance of the tissue type used to detect pathogens. PMID:24484498

  10. Antimicrobial, antitumor and brine shrimp lethality assay of Ranunculus arvensis L. extracts.

    Science.gov (United States)

    Bhatti, Muhammad Zeeshan; Ali, Amjad; Saeed, Asma; Saeed, Ahmad; Malik, Salman Akbar

    2015-05-01

    To investigate the antitumor activity, brine shrimp lethality assay, antibacterial and antifungal activity of Methanol Extract (ME), Water Extract (WE), Acetone Extract (AE), Chloroform Extract (CE), Methanol-Water Extract (MWE), Methanol-Acetone Extract (MAE), Methanol-Chloroform Extract (MCE) of Ranunculus arvensis (L.). Antitumor activity was evaluated with Agrobacterium tumefaciens (At10) induced potato disc assay. Cytotoxicity was evaluated with brine shrimp lethality assay. Antibacterial activity was evaluated with six bacterial strains including Escherichia coli, Enterobacter aerogenes, Bordetella bronchiseptica, Klebsiella pneumoniae, Micrococcus luteus and Streptococcus anginosus and antifungal screening was done against five fungal strains including Aspergillus niger, A. flavus, A. fumigates, Fusarium solani and Mucor species by using disc diffusion method. Best antitumor activity was obtained with ME and WE, having highest IC50 values 20.27 ± 1.62 and 93.01 ± 1.33μg/disc. Brine shrimp lethality assay showed LC50 values of AE, MAE and ME were obtained as 384.66 ± 9.42μg/ml, 724.11 ± 8.01μg/ml and 978.7 ±8.01 μg/ml respectively. WE of R. arvensis revealed weak antimicrobial result against the tested microorganisms. On the other hand, the antifungal activity of the plant extracts was found to be insignificant. These findings demonstrate that extracts of R. arvensis possesses significant antitumor activity. Further extensive study is necessary to assess the therapeutic potential of the plant. PMID:26004705

  11. Role of Atypical Pathogens and the Antibiotic Prescription Pattern in Acute Bronchitis: A Multicenter Study in Korea.

    Science.gov (United States)

    Park, Sunghoon; Oh, Kil Chan; Kim, Ki-Seong; Song, Kyu-Tae; Yoo, Kwang Ha; Shim, Yun Su; Lee, Young Ju; Lee, Myung Goo; Yun, Jang Uk; Kim, Hyun Su; Kim, Yee Hyung; Lee, Won Jun; Kim, Do Il; Cha, Hyung Gun; Lee, Jae-Myung; Seo, Jung San; Jung, Ki-Suck

    2015-10-01

    The role of atypical bacteria and the effect of antibiotic treatments in acute bronchitis are still not clear. This study was conducted at 22 hospitals (17 primary care clinics and 5 university hospitals) in Korea. Outpatients (aged ≥ 18 yr) who had an acute illness with a new cough and sputum (≤ 30 days) were enrolled in 2013. Multiplex real-time polymerase chain reaction (RT-PCR) was used to detect five atypical bacteria. A total of 435 patients were diagnosed as having acute bronchitis (vs. probable pneumonia, n = 75), and 1.8% (n = 8) were positive for atypical pathogens (Bordetella pertussis, n = 3; B. parapertussis, n = 0; Mycoplasma pneumoniae, n = 1; Chlamydophila pneumoniae, n = 3; Legionella pneumophila, n = 1). Among clinical symptoms and signs, only post-tussive vomiting was more frequent in patients with atypical pathogens than those without (P = 0.024). In all, 72.2% of the enrolled patients received antibiotic treatment at their first visits, and β-lactams (29.4%) and quinolones (20.5%) were the most commonly prescribed agents. In conclusion, our study demonstrates that the incidence of atypical pathogens is low in patients with acute bronchitis, and the rate of antibiotic prescriptions is high. PMID:26425041

  12. The spread of non-OIE-listed avian diseases through international trade of chicken meat: an assessment of the risks to New Zealand.

    Science.gov (United States)

    Cobb, S P; Smith, H

    2015-12-01

    Twelve avian diseases are listed by the World Organisation for Animal Health (OIE), although more than 100 infectious diseases have been described in commercial poultry. This article summarises a recent assessment of the biosecurity risks posed by non-listed avian diseases associated with imports of chilled or frozen chicken meat and meat products into New Zealand. Following the guidelines described in Chapter 2.1 of the OIE Terrestrial Animal Health Code, avian adenovirus splenomegaly virus, avian paramyxovirus-2 (APMV-2), Bordetella avium, Mycoplasma spp., Ureaplasma spp., Ornithobacterium rhinotracheale, Riemerella anatipestifer, and Salmonella arizonae have been identified as hazards. However, of all the non-listed avian diseases discussed here, only APMV-2 and S. arizonae are assessed as being risks associated with the commercial import of chicken meat into New Zealand. Specific control measures may have to be implemented to mitigate such risks. This conclusion is likely to reflect both the high-health status of New Zealand poultry and the threat posed by these infectious agents to New Zealand's unique population of native psittacine species. PMID:27044152

  13. Enhanced crude oil biodegradation in soil via biostimulation.

    Science.gov (United States)

    Al-Saleh, Esmaeil; Hassan, Ali

    2016-08-01

    Research on feasible methods for the enhancement of bioremediation in soil contaminated by crude oil is vital in oil-exporting countries such as Kuwait, where crude oil is a major pollutant and the environment is hostile to biodegradation. This study investigated the possibility of enhancing crude oil bioremediation by supplementing soil with cost-effective organic materials derived from two widespread locally grown trees, Conocarpus and Tamarix. Amendments in soils increased the counts of soil microbiota by up to 98% and enhanced their activity by up to 95.5%. The increase in the biodegradation of crude oil (75%) and high levels of alkB expression substantiated the efficiency of the proposed amendment technology for the bioremediation of hydrocarbon-contaminated sites. The identification of crude-oil-degrading bacteria revealed the dominance of the genus Microbacterium (39.6%), Sphingopyxis soli (19.3%), and Bordetella petrii (19.6%) in unamended, Conocarpus-amended, and Tamarix-amended contaminated soils, respectively. Although soil amendments favored the growth of Gram-negative bacteria and reduced bacterial diversity, the structures of bacterial communities were not significantly altered. PMID:26854134

  14. BACTERIAL FLORA OF HATCHERY ENVIRONMENT AND THEIR IN-VITRO SUSCEPTIBILITY TO ANTIMICROBIAL AGENTS

    Directory of Open Access Journals (Sweden)

    F. M. Khan, H. Afzal and F. Deeba

    2003-04-01

    Full Text Available Four hatcheries, located in and around Faisalabad, were sampled a day before hatch out in six batches for environmental bacterial flora. Hatchery air, egg-shell surface, surfaces of selected locations and water supply samples were taken for this purpose. The percent (relative occurrence of various bacterial species recovered from hatchery environment revealed that Bacillus subtilis was the predominant isolate (26.93%. followed by Escherichia coli (24.08%, Staphylococcus epidermidis (16.32%, Staphylococcus aureus (8.16%, Paratyphoid salmonellae (6.93%, Pseudomonas aeruginosa (4.48%, Citrobacter jreundii (4.08%, Enterococcus faecalis (3.26%, Klebsiella pneumoniae (3.26%, Bordetella avium (1.63% and Proteus vulgaris (0.81%. In second part of the study, bacterial isolates were subjected to in-vitro antibiotic sensitivity to 8 antibiotics of common poultry use. It was found that 98.92, 79.56. 65.59, 61.29, 61.29, 61.29, 53.76 and 38.70 percent of bacterial isolates were sensitive to Norfloxacin, Gentamicin, Neomycin, Chloramphenicol, Doxycycline, Flumequine, Erythromycin, and Ampicillin, respectively. In the final part of the study, bacterial isolates were tested for resistance to 3 commerical hatchery disinfectants (TH4®, Aldekol Des® 0.2, and Bromosept 10% soln. ®. Only 3.22% of the isolates showed resistance at manufacturer's recommended dilution (MRD levels while 11.82% of the isolates showed resistance at concentrations below the MRD levels.

  15. Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling.

    Directory of Open Access Journals (Sweden)

    Mariette Barbier

    Full Text Available Since their discovery, fluorescent proteins have been widely used to study protein function, localization or interaction, promoter activity and regulation, drug discovery or for non-invasive imaging. They have been extensively modified to improve brightness, stability, and oligomerization state. However, only a few studies have focused on understanding the dynamics of fluorescent proteins expression in bacteria. In this work, we developed a set plasmids encoding 12 fluorescent proteins for bacterial labeling to facilitate the study of pathogen-host interactions. These broad-spectrum plasmids can be used with a wide variety of Gram-negative microorganisms including Escherichia coli, Pseudomonas aeruginosa, Burkholderia cepacia, Bordetella bronchiseptica, Shigella flexneri or Klebsiella pneumoniae. For comparison, fluorescent protein expression and physical characteristics in Escherichia coli were analyzed using fluorescence microscopy, flow cytometry and in vivo imaging. Fluorescent proteins derived from the Aequorea Victoria family showed high photobleaching, while proteins form the Discosoma sp. and the Fungia coccina family were more photostable for microscopy applications. Only E2-Crimson, mCherry and mKeima were successfully detected for in vivo applications. Overall, E2-Crimson was the fastest maturing protein tested in E. coli with the best overall performance in the study parameters. This study provides a unified comparison and comprehensive characterization of fluorescent protein photostability, maturation and toxicity, and offers general recommendations on the optimal fluorescent proteins for in vitro and in vivo applications.

  16. Negatively charged residues of the segment linking the enzyme and cytolysin moieties restrict the membrane-permeabilizing capacity of adenylate cyclase toxin

    Science.gov (United States)

    Masin, Jiri; Osickova, Adriana; Sukova, Anna; Fiser, Radovan; Halada, Petr; Bumba, Ladislav; Linhartova, Irena; Osicka, Radim; Sebo, Peter

    2016-01-01

    The whooping cough agent, Bordetella pertussis, secretes an adenylate cyclase toxin-hemolysin (CyaA) that plays a crucial role in host respiratory tract colonization. CyaA targets CR3-expressing cells and disrupts their bactericidal functions by delivering into their cytosol an adenylate cyclase enzyme that converts intracellular ATP to cAMP. In parallel, the hydrophobic domain of CyaA forms cation-selective pores that permeabilize cell membrane. The invasive AC and pore-forming domains of CyaA are linked by a segment that is unique in the RTX cytolysin family. We used mass spectrometry and circular dichroism to show that the linker segment forms α-helical structures that penetrate into lipid bilayer. Replacement of the positively charged arginine residues, proposed to be involved in target membrane destabilization by the linker segment, reduced the capacity of the toxin to translocate the AC domain across cell membrane. Substitutions of negatively charged residues then revealed that two clusters of negative charges within the linker segment control the size and the propensity of CyaA pore formation, thereby restricting the cell-permeabilizing capacity of CyaA. The ‘AC to Hly-linking segment’ thus appears to account for the smaller size and modest cell-permeabilizing capacity of CyaA pores, as compared to typical RTX hemolysins. PMID:27581058

  17. The cis-state of an azobenzene photoswitch is stabilized through specific interactions with a protein surface.

    Science.gov (United States)

    Korbus, Michael; Backé, Sarah; Meyer-Almes, Franz-Josef

    2015-03-01

    The photocontrol of protein function like enzyme activity has been the subject of many investigations to enable reversible and spatiotemporally defined cascading biochemical reactions without the need for separation in miniaturized and parallelized assay setups for academic and industrial applications. A photoswitchable amidohydrolase variant from Bordetella/Alcaligenes with the longest reported half-life (approximately 30 h) for the cis-state of the attached azobenzene group was chosen as a model system to dissect the underlying mechanism and molecular interactions that caused the enormous deceleration of the thermal cis-to-trans relaxation of the azobenzene photoswitch. A systematic site-directed mutagenesis study on the basis of molecular dynamics simulation data was employed to investigate enzyme and thermal cis-to-trans relaxation kinetics in dependence on selected amino acid substitution, which revealed a prominent histidine and a hydrophobic cluster as molecular determinants for the stabilization of the cis-isomer of the attached azobenzene moiety on the protein surface. The nature of the involved interactions consists of polar, hydrophobic, and possibly aromatic Π-Π contributions. The elucidated principles behind the stabilization of the cis-state of azobenzene derivatives on a protein surface can be exploited to design improved biologically inspired photoswitches. Moreover, the findings open the door to highly long-lived cis-states of azobenzene groups yielding improved bistable photoswitches that can be controlled by single light-pulses rather than continuous irradiation with UV light that causes potential photodamage to the employed biomolecules. PMID:25664524

  18. MICROBIAL POPULATION OF HOT SPRING WATERS IN ESKİŞEHİR/TURKEY

    Directory of Open Access Journals (Sweden)

    Nalan YILMAZ SARIÖZLÜ

    2012-02-01

    Full Text Available In order to investigate and find out the bacterial community of hot spring waters in Eskişehir, Turkey, 7 hot spring water samples were collected from 7 different hot springs. All samples were inoculated using four different media (nutrient agar, water yeast extract agar, trypticase soy agar, starch casein agar. After incubation at 50 ºC for 14 days, all bacterial colonies were counted and purified. Gram reaction, catalase and oxidase properties of all isolates were determined and investigated by BIOLOG, VITEK and automated ribotyping system (RiboPrinter. The resistance of these bacteriawas examined against ampiciline, gentamisine, trimethoprime-sulphamethoxazole and tetracycline. As a result, heat resistant pathogenic microorganisms in addition to human normal flora were determined in hot spring waters (43-50 ºC in investigated area. Ten different species belong to 6 genera were identified as Alysiella filiformis, Bordetella bronchiseptica, B. pertussis, Molexalla caprae, M. caviae, M. cuniculi, M. phenylpyruvica, Roseomonas fauriae, Delftia acidovorans and Pseudomonas taetrolens.

  19. Treatment with Vitamin D/MOG Association Suppresses Experimental Autoimmune Encephalomyelitis.

    Directory of Open Access Journals (Sweden)

    Fernanda Chiuso-Minicucci

    Full Text Available Experimental autoimmune encephalomyelitis (EAE is an animal model to study multiple sclerosis (MS. Considering the tolerogenic effects of active vitamin D, we evaluated the therapeutic effect of myelin oligodendrocyte glycoprotein (MOG associated with active vitamin D in EAE development. EAE was induced in female C57BL/6 mice by immunization with MOG emulsified with Complete Freund's Adjuvant plus Mycobacterium tuberculosis. Animals also received two intraperitoneal doses of Bordetella pertussis toxin. One day after immunization, mice were treated with 0,1 μg of 1α,25-dihydroxyvitamin D3 (1,25(OH2D3 every other day during 15 days (on days 1, 3, 5, 7, 9, 11, 13 and 15. MOG (150 μg was co-administered on days 3 and 11. The administration of 1,25(OH2D3 or MOG determined significant reduction in EAE incidence and in clinical scores. When MOG was associated with 1,25(OH2D3 the animals did not develop EAE. Spleen and central nervous system (CNS cell cultures from this group produced less IL-6 and IL-17 upon stimulation with MOG in comparison to the EAE control group. In addition, this treatment inhibited dendritic cells maturation in the spleen and reduced inflammatory infiltration in the CNS. The association of MOG with 1,25(OH2D3 was able to control EAE development.

  20. Isolation and characterization of facultative mixotrophic ammonia-oxidizing bacteria from constructed wetlands

    Institute of Scientific and Technical Information of China (English)

    Soulwène Kouki; Neila Saidi; Fadhel M'hiri; Houda Nasr; Hanène Cherif; Hadda Ouzari; Abdermaceur Hassen

    2011-01-01

    Autotrophic ammonia-oxidizing bacteria (AOB) have been widely studied in constructed wetlands systems,while mixotrophic AOB have been less thoroughly examined.Heterotrophic bacteria were isolated from wastewater and rhizospheres of macrophytes of constructed wetlands,and then cultivated in a mixotrophic medium containing ammonium and acetic acid.A molecular characterization was accomplished using ITS-PCR amplification,and phylogenetic analysis based on 16S rRNA gene sequences.Results showed the presence of 35 bacteria,among 400 initially heterotrophic isolates,that were able to remove ammonia.These 35 isolates were classified into 10 genetically different groups based on ITS pattern.Then,a collection of 10 isolates were selected because of their relatively high ammonia removal efficiencies (ARE ≥ 80%) and their phylogenetic diversity.In conditions of mixotrophy,these strains were shown to be able to grow (increase of optical density OD660 during incubation with assimilation of nitrogen into cellular biomass) and to oxidize ammonia (important ammonia oxidation efficiencies,AOE between 79% and 87%).Among these facultative mixotrophic AOB,four isolates were genetically related to Firmicutes (Bacillus and Exiguobacterium),three isolates were affiliated to Actinobacteria (Arthrobacter) and three other isolates were associated with Proteobacteria (Pseudomonas,Ochrobactrum and Bordetella).

  1. The potential of adjuvants to improve immune responses against TdaP vaccines: A preclinical evaluation of MF59 and monophosphoryl lipid A.

    Science.gov (United States)

    Agnolon, Valentina; Bruno, Cristina; Leuzzi, Rosanna; Galletti, Bruno; D'Oro, Ugo; Pizza, Mariagrazia; Seubert, Anja; O'Hagan, Derek T; Baudner, Barbara C

    2015-08-15

    The successful approach of combining diphtheria, tetanus and pertussis antigens into a single vaccine has become a cornerstone of immunization programs. Yet, even if vaccination coverage is high, a resurgence of pertussis has been reported in many countries suggesting current vaccines may not provide adequate protection. To induce better tailored and more durable immune responses against pertussis vaccines different approaches have been proposed, including the use of novel adjuvants. Licensed aP vaccines contain aluminum salts, which mainly stimulate humoral immune responses and might not be ideal for protecting against Bordetella pertussis infection. Adjuvants inducing more balanced T-helper profiles or even Th1-prone responses might be more adequate. In this study, two adjuvants already approved for human use have been tested: MF59 emulsion and the combination of aluminum hydroxide with the Toll-Like Receptor 4 agonist MPLA. Adjuvanticity was evaluated in a mouse model using a TdaP vaccine containing three B. pertussis antigens: genetically detoxified pertussis toxin (PT-9K/129G), filamentous hemagglutinin (FHA) and pertactin (PRN) The physico-chemical compatibility of TdaP antigens with the proposed adjuvants, together with a quicker onset and changed quality of the antibody responses, fully supports the replacement of aluminum salts with a new adjuvant to enhance aP vaccines immunogenicity. PMID:26149936

  2. Dysregulated microRNA Expression in Serum of Non-Vaccinated Children with Varicella

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    Yuhua Qi

    2014-04-01

    Full Text Available Circulating microRNAs (miRNAs may play an important role in pathogen-host interactions and can serve as molecular markers for the detection of infectious diseases. To date, the relationship between circulating miRNAs and varicella-zoster virus (VZV caused varicella has not been reported. Using TaqMan Low-Density Array (TLDA analysis, expression levels of miRNAs in serum samples from 29 patients with varicella and 60 patients with Bordetella pertussis (BP, measles virus (MEV and enterovirus (EV were analyzed. The array results showed that 247 miRNAs were differentially expressed in sera of the varicella patients compared with healthy controls (215 up-regulated and 32 down-regulated. Through the following qRT-PCR confirmation and receiver operational characteristic (ROC curve analysis, five miRNAs (miR-197, miR-629, miR-363, miR-132 and miR-122 were shown to distinguish varicella patients from healthy controls and other microbial infections with moderate sensitivity and specificity. A number of significantly enriched pathways regulated by these circulating miRNAs were predicted, and some of them were involved in inflammatory response, nervous system and respiratory system development. Our results, for the first time, revealed that a number of miRNAs were differentially expressed during VZV infection, and these five serum miRNAs have great potential to serve as biomarkers for the diagnosis of VZV infection in varicella patients.

  3. Quantitative Mass Spectrometry for Bacterial Protein Toxins — A Sensitive, Specific, High-Throughput Tool for Detection and Diagnosis

    Directory of Open Access Journals (Sweden)

    Suzanne Kalb

    2011-03-01

    Full Text Available Matrix-assisted laser-desorption time-of-flight (MALDI-TOF mass spectrometry (MS is a valuable high-throughput tool for peptide analysis. Liquid chromatography electrospray ionization (LC-ESI tandem-MS provides sensitive and specific quantification of small molecules and peptides. The high analytic power of MS coupled with high-specificity substrates is ideally suited for detection and quantification of bacterial enzymatic activities. As specific examples of the MS applications in disease diagnosis and select agent detection, we describe recent advances in the analyses of two high profile protein toxin groups, the Bacillus anthracis toxins and the Clostridium botulinum neurotoxins. The two binary toxins produced by B. anthracis consist of protective antigen (PA which combines with lethal factor (LF and edema factor (EF, forming lethal toxin and edema toxin respectively. LF is a zinc-dependent endoprotease which hydrolyzes specific proteins involved in inflammation and immunity. EF is an adenylyl cyclase which converts ATP to cyclic-AMP. Toxin-specific enzyme activity for a strategically designed substrate, amplifies reaction products which are detected by MALDI-TOF-MS and LC-ESI-MS/MS. Pre-concentration/purification with toxin specific monoclonal antibodies provides additional specificity. These combined technologies have achieved high specificity, ultrasensitive detection and quantification of the anthrax toxins. We also describe potential applications to diseases of high public health impact, including Clostridium difficile glucosylating toxins and the Bordetella pertussis adenylyl cyclase.

  4. Pharmacophore Selection and Redesign of Non-nucleotide Inhibitors of Anthrax Edema Factor

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    Maria Estrella Jimenez

    2012-11-01

    Full Text Available Antibiotic treatment may fail to protect individuals, if not started early enough, after infection with Bacillus anthracis, due to the continuing activity of toxins that the bacterium produces. Stable and easily stored inhibitors of the edema factor toxin (EF, an adenylyl cyclase, could save lives in the event of an outbreak, due to natural causes or a bioweapon attack. The toxin’s basic activity is to convert ATP to cAMP, and it is thus in principle a simple phosphatase, which means that many mammalian enzymes, including intracellular adenylcyclases, may have a similar activity. While nucleotide based inhibitors, similar to its natural substrate, ATP, were identified early, these compounds had low activity and specificity for EF. We used a combined structural and computational approach to choose small organic molecules in large, web-based compound libraries that would, based on docking scores, bind to residues within the substrate binding pocket of EF. A family of fluorenone-based inhibitors was identified that inhibited the release of cAMP from cells treated with EF. The lead inhibitor was also shown to inhibit the diarrhea caused by enterotoxigenic E. coli (ETEC in a murine model, perhaps by serving as a quorum sensor. These inhibitors are now being tested for their ability to inhibit Anthrax infection in animal models and may have use against other pathogens that produce toxins similar to EF, such as Bordetella pertussis or Vibrio cholera.

  5. Evaluation of adjuvant effects of fucoidan for improving vaccine efficacy.

    Science.gov (United States)

    Kim, Su-Yeon; Joo, Hong-Gu

    2015-01-01

    Fucoidan is a sulfated polysaccharide derived from brown seaweed, including Fucus vesiculosus. This compound is known to have immunostimulatory effects on various types of immune cells including macrophages and dendritic cells. A recent study described the application of fucoidan as a vaccine adjuvant. Vaccination is regarded as the most efficient prophylactic method for preventing harmful or epidemic diseases. To increase vaccine efficacy, effective adjuvants are needed. In the present study, we determined whether fucoidan can function as an adjuvant using vaccine antigens. Flow cytometric analysis revealed that fucoidan increases the expression of the activation markers major histocompatibility complex class II, cluster of differentiation (CD)25, and CD69 in spleen cells. In combination with Bordetella bronchiseptica antigen, fucoidan increased the viability and tumor necrosis factor-α production of spleen cells. Furthermore, fucoidan increased the in vivo production of antigen-specific antibodies in mice inoculated with Mycoplasma hyopneumoniae antigen. Overall, this study has provided valuable information about the use of fucoidan as a vaccine adjuvant. PMID:25549218

  6. Limitaciones del ensayo de toxicidad específica para el componente pertussis de células completas

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    Vicente Perdomo

    2008-01-01

    Full Text Available Las vacunas que contienen células inactivadas de Bordetella pertussis se han utilizado con efectividad en los Programas Nacionales de Inmunización de todo el mundo. Pese a su reconocida eficacia, ellas se caracterizan por su elevada reactogenicidad, atribuible a la presencia de componentes como toxina pertussis y endotoxinas. Paramonitorear la seguridad de estas vacunas existe el ensayo de ganancia en peso en ratones, el cual ha sido criticado por su inespecificidad, poca sensibilidad y alta variabilidad. Basado en lo anterior, el Laboratorio Nacional de Biológicos del Centro Estatal para el Control de de la Calidad de los Medicamentos en Cuba, decidió evaluar la relevancia de esta prueba para la liberación de los lotes de la vacuna DPT. Para ello se estimó la sensibilidad del método para detectar diferentes concentraciones de endotoxinas y toxina pertussis, así como la variabilidad entre ensayos. Los resultados de este trabajo mostraron que sólo altas concentraciones de endotoxinas y toxina pertussis, muy superiores a las habituales en las vacunas DPT, provocan una disminución de la ganancia en peso promedio y un fallo en la especificación de la prueba. Este elemento y la inherente variabilidad de este método resultaron claves en la decisión de no utilizarlo para la liberación nacional de lotes de la vacuna DPT.

  7. Limitaciones del ensayo de toxicidad específica para el componente pertussis de células completas

    Directory of Open Access Journals (Sweden)

    Mario Landys Chovel

    2008-12-01

    Full Text Available Las vacunas que contienen células inactivadas de Bordetella pertussis se han utilizado con efectividad en los Programas Nacionales de Inmunización de todo el mundo. Pese a su reconocida eficacia, ellas se caracterizan por su elevada reactogenicidad, atribuible a la presencia de componentes como toxina pertussis y endotoxinas. Para monitorear la seguridad de estas vacunas existe el ensayo de ganancia en peso en ratones, el cual ha sido criticado por su inespecificidad, poca sensibilidad y alta variabilidad. Basado en lo anterior, el Laboratorio Nacional de Biológicos del Centro Estatal para el Control de de la Calidad de los Medicamentos en Cuba, decidió evaluar la relevancia de esta prueba para la liberación de los lotes de la vacuna DPT. Para ello se estimó la sensibilidad del método para detectar diferentes concentraciones de endotoxinas y toxina pertussis, así como la variabilidad entre ensayos. Los resultados de este trabajo mostraron que sólo altas concentraciones de endotoxinas y toxina pertussis, muy superiores a las habituales en las vacunas DPT, provocan una disminución de la ganancia en peso promedio y un fallo en la especificación de la prueba. Este elemento y la inherente variabilidad de este método resultaron claves en la decisión de no utilizarlo para la liberación nacional de lotes de la vacuna DPT.

  8. A retrospective study of acute pertussis in Hasan Sadikin Hospital-Indonesia

    Institute of Scientific and Technical Information of China (English)

    Heda Melinda Nataprawira; Evelyn Phangkawira

    2015-01-01

    Objective: To describe the representation of pertussis diagnosis in children. Methods: A retrospective observational study was performed on pediatric pertussis and pertussis-like syndrome registry for children <14 years of age documented from October 2008 to December 2014 in Hasan Sadikin Hospital, Indonesia. Demographic data, signs and symptoms at presentation, case definition (probable, confirmed), possible pertussis contact, pertussis vaccination status, results of Bordetella pertussis (B. pertussis) culture, complications, and outcome were recorded. Results:Sixty-one probable and two confirmed pertussis were documented. Male and female ratio was 1:1, mostly presented with shortness of breath, 24 (38%) subjects had posttussive vomiting, 10 (16%) had whooping-cough. Ten patients (16%) were reported to have adult possible pertussis contact. Only 2 infants had previous pertussis vaccination. All subjects presented in the second week of illness were all diagnosed as bronchopneumonia but two. The mean age was 6 months, ranging from 0−50 months. One subject required mechanical ventilation. B. pertussis culture was performed only in 35 (56%) subjects but positive only in two. There were no fatal cases, 55 (87%) including the subject who need mechanical ventilation had good outcome. Conclusions: Mostly patients were admitted on paroxysmal phase when no more active B. pertussis could be found from nasopharyngeal secret. A rigorous history taking particularly excessive cough, posttussive vomitting, and pertussis vaccination status need to be taken into account.

  9. Response of soybean rhizosphere communities to human hygiene water addition as determined by community level physiological profiling (CLPP) and terminal restriction fragment length polymorphism (TRFLP) analysis

    Science.gov (United States)

    Kerkhof, L.; Santoro, M.; Garland, J.

    2000-01-01

    In this report, we describe an experiment conducted at Kennedy Space Center in the biomass production chamber (BPC) using soybean plants for purification and processing of human hygiene water. Specifically, we tested whether it was possible to detect changes in the root-associated bacterial assemblage of the plants and ultimately to identify the specific microorganism(s) which differed when plants were exposed to hygiene water and other hydroponic media. Plants were grown in hydroponics media corresponding to four different treatments: control (Hoagland's solution), artificial gray water (Hoagland's+surfactant), filtered gray water collected from human subjects on site, and unfiltered gray water. Differences in rhizosphere microbial populations in all experimental treatments were observed when compared to the control treatment using both community level physiological profiles (BIOLOG) and molecular fingerprinting of 16S rRNA genes by terminal restriction fragment length polymorphism analysis (TRFLP). Furthermore, screening of a clonal library of 16S rRNA genes by TRFLP yielded nearly full length SSU genes associated with the various treatments. Most 16S rRNA genes were affiliated with the Klebsiella, Pseudomonas, Variovorax, Burkholderia, Bordetella and Isosphaera groups. This molecular approach demonstrated the ability to rapidly detect and identify microorganisms unique to experimental treatments and provides a means to fingerprint microbial communities in the biosystems being developed at NASA for optimizing advanced life support operations.

  10. A fully integrated microfluidic genetic analysis system with sample-in-answer-out capability.

    Science.gov (United States)

    Easley, Christopher J; Karlinsey, James M; Bienvenue, Joan M; Legendre, Lindsay A; Roper, Michael G; Feldman, Sanford H; Hughes, Molly A; Hewlett, Erik L; Merkel, Tod J; Ferrance, Jerome P; Landers, James P

    2006-12-19

    We describe a microfluidic genetic analysis system that represents a previously undescribed integrated microfluidic device capable of accepting whole blood as a crude biological sample with the endpoint generation of a genetic profile. Upon loading the sample, the glass microfluidic genetic analysis system device carries out on-chip DNA purification and PCR-based amplification, followed by separation and detection in a manner that allows for microliter samples to be screened for infectious pathogens with sample-in-answer-out results in pump delivers sample/reagents to the chip for nucleic acid purification from a biological sample. Elastomeric membrane valving isolates each distinct functional region of the device and, together with resistive flow, directs purified DNA and PCR reagents from the extraction domain into a 550-nl chamber for rapid target sequence PCR amplification. Repeated pressure-based injections of nanoliter aliquots of amplicon (along with the DNA sizing standard) allow electrophoretic separation and detection to provide DNA fragment size information. The presence of Bacillus anthracis (anthrax) in 750 nl of whole blood from living asymptomatic infected mice and of Bordetella pertussis in 1 microl of nasal aspirate from a patient suspected of having whooping cough are confirmed by the resultant genetic profile. PMID:17159153

  11. Pertussis outbreak in Papua New Guinea: the challenges of response in a remote geo-topographical setting

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    William Lagani

    2012-10-01

    Full Text Available Introduction: A large outbreak of pertussis was detected during March 2011 in Goilala, a remote district of the Central Province in Papua New Guinea, characterized by rugged topography with no road access from the provincial headquarters. This outbreak investigation highlights the difficulties in reporting and responding to outbreaks in these settings.Method: The suspected pertussis cases, reported by health workers from the Ononge health centre area, were investigated and confirmed for the presence of Bordetella pertussis DNA using the polymerase chain reaction (PCR method.Results: There were 205 suspected pertussis cases, with a case-fatality rate (CFR of 3%. All cases were unvaccinated. The Central Province conducted a response vaccination programme providing 65% of children less than five years of age with diphtheria–pertussis-tetanus-HepB-Hib vaccine at a cost of US$ 12.62 per child.Discussion: The incurred cost of vaccination in response to this outbreak was much higher than the US$ 3.80 per child for routine outreach patrol. To prevent further outbreaks of vaccine-preventable diseases in these areas, local health centres must ensure routine vaccination is strengthened through the “Reaching Every District” initiative of the National Department of Health.

  12. Photolabeling of Glu-129 of the S-1 subunit of pertussis toxin with NAD

    Energy Technology Data Exchange (ETDEWEB)

    Barbieri, J.T.; Mende-Mueller, L.M.; Rappuoli, R.; Collier, R.J. (Medical College of Wisconsin, Milwaukee (USA))

    1989-11-01

    UV irradiation was shown to induce efficient transfer of radiolabel from nicotinamide-labeled NAD to a recombinant protein (C180 peptide) containing the catalytic region of the S-1 subunit of pertussis toxin. Incorporation of label from (3H-nicotinamide)NAD was efficient (0.5 to 0.6 mol/mol of protein) relative to incorporation from (32P-adenylate)NAD (0.2 mol/mol of protein). Label from (3H-nicotinamide)NAD was specifically associated with Glu-129. Replacement of Glu-129 with glycine or aspartic acid made the protein refractory to photolabeling with (3H-nicotinamide)NAD, whereas replacement of a nearby glutamic acid, Glu-139, with serine did not. Photolabeling of the C180 peptide with NAD is similar to that observed with diphtheria toxin and exotoxin A of Pseudomonas aeruginosa, in which the nicotinamide portion of NAD is transferred to Glu-148 and Glu-553, respectively, in the two toxins. These results implicate Glu-129 of the S-1 subunit as an active-site residue and a potentially important site for genetic modification of pertussis toxin for development of an acellular vaccine against Bordetella pertussis.

  13. DNA-binding studies and biological activities of new nitrosubstituted acyl thioureas

    Science.gov (United States)

    Tahir, Shaista; Badshah, Amin; Hussain, Raja Azadar; Tahir, Muhammad Nawaz; Tabassum, Saira; Patujo, Jahangir Ali; Rauf, Muhammad Khawar

    2015-11-01

    Four new nitrosubstituted acylthioureas i.e. 1-acetyl-3-(4-nitrophenyl)thiourea (TU1), 1-acetyl-3-(2-methyl-4-nitrophenyl)thiourea (TU2), 1-acetyl-3-(2-methoxy-4-nitrophenyl)thiourea (TU3) and 1-acetyl-3-(4-chloro-3-nitrophenyl)thiourea (TU4) have been synthesized and characterized (by C13 and H1 nuclear magnetic resonance, Fourier transform infrared spectroscopy and single crystal X-ray diffraction). As a preliminary investigation of the anti-cancer potencies of the said compounds, DNA interaction studies have been carried out using cyclic voltammetry and UV-vis spectroscopy along with verification from computational studies. The drug-DNA binding constants are found to be in the order, KTU3 9.04 × 106 M-1 > KTU4 8.57 × 106 M-1 > KTU2 6.05 × 106 M-1 > KTU1 1.16 × 106 M-1. Furthermore, the antioxidant, cytotoxic, antibacterial and antifungal activities have been carried out against DPPH (1,1-diphenyl-2-dipicrylhydrazyl), Brine shrimp eggs, gram positive (Micrococcus luteus, Staphylococcus aureus) and gram negative (Bordetella bronchiseptica, Salmonella typhimurium, Enterobacter aerogens) and fungal cultures (Aspergillus fumigatus, Mucor species, Aspergillus niger, Aspergillus flavus) respectively.

  14. Complex structure of a bacterial class 2 histone deacetylase homologue with a trifluoromethylketone inhibitor

    International Nuclear Information System (INIS)

    The crystal structure of HDAH FB188 in complex with a trifluoromethylketone at 2.2 Å resolution is reported and compared to a previously determined inhibitor complex. Histone deacetylases (HDACs) have emerged as attractive targets in anticancer drug development. To date, a number of HDAC inhibitors have been developed and most of them are hydroxamic acid derivatives, typified by suberoylanilide hydroxamic acid (SAHA). Not surprisingly, structural information that can greatly enhance the design of novel HDAC inhibitors is so far only available for hydroxamic acids in complex with HDAC or HDAC-like enzymes. Here, the first structure of an enzyme complex with a nonhydroxamate HDAC inhibitor is presented. The structure of the trifluoromethyl ketone inhibitor 9,9,9-trifluoro-8-oxo-N-phenylnonanamide in complex with bacterial FB188 HDAH (histone deacetylase-like amidohydrolase from Bordetella/Alcaligenes strain FB188) has been determined. HDAH reveals high sequential and functional homology to human class 2 HDACs and a high structural homology to human class 1 HDACs. Comparison with the structure of HDAH in complex with SAHA reveals that the two inhibitors superimpose well. However, significant differences in binding to the active site of HDAH were observed. In the presented structure the O atom of the trifluoromethyl ketone moiety is within binding distance of the Zn atom of the enzyme and the F atoms participate in interactions with the enzyme, thereby involving more amino acids in enzyme–inhibitor binding

  15. Calcium-Driven Folding of RTX Domain β-Rolls Ratchets Translocation of RTX Proteins through Type I Secretion Ducts.

    Science.gov (United States)

    Bumba, Ladislav; Masin, Jiri; Macek, Pavel; Wald, Tomas; Motlova, Lucia; Bibova, Ilona; Klimova, Nela; Bednarova, Lucie; Veverka, Vaclav; Kachala, Michael; Svergun, Dmitri I; Barinka, Cyril; Sebo, Peter

    2016-04-01

    Calcium-binding RTX proteins are equipped with C-terminal secretion signals and translocate from the Ca(2+)-depleted cytosol of Gram-negative bacteria directly into the Ca(2+)-rich external milieu, passing through the "channel-tunnel" ducts of type I secretion systems (T1SSs). Using Bordetella pertussis adenylate cyclase toxin, we solved the structure of an essential C-terminal assembly that caps the RTX domains of RTX family leukotoxins. This is shown to scaffold directional Ca(2+)-dependent folding of the carboxy-proximal RTX repeat blocks into β-rolls. The resulting intramolecular Brownian ratchets then prevent backsliding of translocating RTX proteins in the T1SS conduits and thereby accelerate excretion of very large RTX leukotoxins from bacterial cells by a vectorial "push-ratchet" mechanism. Successive Ca(2+)-dependent and cosecretional acquisition of a functional RTX toxin structure in the course of T1SS-mediated translocation, through RTX domain folding from the C-terminal cap toward the N terminus, sets a paradigm that opens for design of virulence inhibitors of major pathogens. PMID:27058787

  16. Epidemiologic evaluation of multiple respiratory pathogens in cats in animal shelters.

    Science.gov (United States)

    Bannasch, Michael J; Foley, Janet E

    2005-04-01

    Upper respiratory tract infection (URI) propagates readily within cats in shelters and often results in euthanasia of affected cats. In a case-control evaluation of 573 cats in eight shelters in California in 2001 and 2002, the prevalence of feline calicivirus (FCV) was from 13 to 36%, feline herpesvirus (FHV) was from 3 to 38%, and prevalence of Bordetella bronchiseptica, Chlamydophila felis, and Mycoplasma species was from 2 to 14%. Cats with URI tended to be housed in isolation, dehydrated, and younger than cats without URI, and infected with FHV, Mycoplasma species, FCV, or C felis. Shelters differed in the prevalence of pathogens and many cats appeared positive for infection after about 1 week of sheltering. It is helpful for shelters to understand the risk factors associated with URI in order to evaluate the costs and benefits of treatment and improve their procedures to decrease the incidence of URI within their facilities. Antiherpetics and antimycoplasmal drugs may be beneficial for individual animal care. Results document the utility of comprehensive URI surveillance and herd management for specific pathogens typical in that shelter. PMID:15771947

  17. Complex structure of a bacterial class 2 histone deacetylase homologue with a trifluoromethylketone inhibitor

    Energy Technology Data Exchange (ETDEWEB)

    Nielsen, Tine Kragh [Abteilung für Molekulare Strukturbiologie, Institut für Mikrobiologie und Genetik and GZMB, Justus-von-Liebig Weg 11, 37077 Göttingen (Germany); Hildmann, Christian; Riester, Daniel; Wegener, Dennis; Schwienhorst, Andreas [Abteilung für Molekulare Genetik und Präparative Molekularbiologie, Institut für Mikrobiologie und Genetik, Grisebachstrasse 8, 37077 Göttingen (Germany); Ficner, Ralf, E-mail: rficner@gwdg.de [Abteilung für Molekulare Strukturbiologie, Institut für Mikrobiologie und Genetik and GZMB, Justus-von-Liebig Weg 11, 37077 Göttingen (Germany)

    2007-04-01

    The crystal structure of HDAH FB188 in complex with a trifluoromethylketone at 2.2 Å resolution is reported and compared to a previously determined inhibitor complex. Histone deacetylases (HDACs) have emerged as attractive targets in anticancer drug development. To date, a number of HDAC inhibitors have been developed and most of them are hydroxamic acid derivatives, typified by suberoylanilide hydroxamic acid (SAHA). Not surprisingly, structural information that can greatly enhance the design of novel HDAC inhibitors is so far only available for hydroxamic acids in complex with HDAC or HDAC-like enzymes. Here, the first structure of an enzyme complex with a nonhydroxamate HDAC inhibitor is presented. The structure of the trifluoromethyl ketone inhibitor 9,9,9-trifluoro-8-oxo-N-phenylnonanamide in complex with bacterial FB188 HDAH (histone deacetylase-like amidohydrolase from Bordetella/Alcaligenes strain FB188) has been determined. HDAH reveals high sequential and functional homology to human class 2 HDACs and a high structural homology to human class 1 HDACs. Comparison with the structure of HDAH in complex with SAHA reveals that the two inhibitors superimpose well. However, significant differences in binding to the active site of HDAH were observed. In the presented structure the O atom of the trifluoromethyl ketone moiety is within binding distance of the Zn atom of the enzyme and the F atoms participate in interactions with the enzyme, thereby involving more amino acids in enzyme–inhibitor binding.

  18. Mechanisms of infection in the respiratory tract.

    Science.gov (United States)

    Baskerville, A

    1981-12-01

    Related to its potential vulnerability the respiratory tract has a very complex and effective defence apparatus. The interaction between these defence mechanisms and certain characteristics of aetiological agents results in a pattern in which initial infections by these agents tend to occur at specific sites in the tract. Infections in which the primary portal of entry is in the upper respiratory tract include Bordetella bronchiseptica and Haemophilus spp in pigs; Pasteurella spp in cattle, sheep, pigs; Mycoplasma spp in cattle, sheep, pigs and poultry; equine herpesvirus 1 in horses; infectious bovine rhinotracheitis in cattle; parainfluenza 3 in cattle and sheep; infectious laryngo-tracheitis and infectious bronchitis in poultry; feline viral rhinotracheitis and calicivirus in cats; Aujeszky's disease virus and swine influenza in pigs; and equine influenza in horses. Infections in which the primary portal of entry is in the lower respiratory tract include Aspergillus fumigatus in poultry and mammals, respiratory syncytial virus in cattle, distemper virus in dogs and adenovirus in cattle and dogs. A fuller understanding of the interactions between an agent and the host at the point of entry would make it much easier to develop effective vaccines and therapeutic agents. PMID:16030806

  19. 猪小肠抗菌肽Cecropin P1对猪呼吸道菌抑菌活性的研究%RESEARCH ON ANTIMICROBIAL ACTIVITY OF CECROPIN P1 TO PORCINE RESPIRATORY BACTERIA

    Institute of Scientific and Technical Information of China (English)

    李伦锋; 马志永; 魏建超; 谢春阳; 彭帅; 李蓓蓓; 刘珂; 邵东华; 邱亚峰; 史子学

    2016-01-01

    Antibacterial peptide cecropin P1 was originally isolated from the small intestines of the pigs. In the present study, the recombinant cecropin P1 produced in pichia pastoris was tested for its antibacterial activity against seven species of common swine respiratory bacteria including Streptococcussuis, Staphylococcusaureus, Pasteurella multocida, Actinobacilluspleuropneumoniae, swine Bordetella bronchiseptica, swine enteropathogenic E.coli and Streptococcus pyogenes. The recombinant cecropin P1 showed different degrees of bacteriostasis for these seven respiratory bacteria as measured by MIC (minimal inhibitory concentration). The most significant bacteristasis was observed for Streptococcus pyogenes, Pasteurella multocida and Staphylococcus aureus with MIC of 49.5 ug/mL, and moderate antibacterial activity for Sreptococcus suis and E.coli. In contrast, the recombinant cecropin P1 had the least bacteriostasis for swine Bordetella bronchiseptica as the MIC was 398 ug/mL. Currently, the vigorous use of antibiotics has caused the bacteria to have developed the high-level resistance. The availability of the recombinant cecropin P1 is of great significance as it possesses wide antimicrobial spectrum, toxin-free nature, high stability, less resistance and no residue in the body.%以猪链球菌、金黄色葡萄球菌、多杀性巴氏杆菌、猪胸膜肺炎放线杆菌、猪支气管败血波氏杆菌、猪致病性大肠杆菌、化脓性链球菌7株猪呼吸道常见菌为实验菌,检测猪小肠抗菌肽对常见的猪呼吸道菌的抑菌效果。结果显示,猪小肠抗菌肽对这7株呼吸道菌均有不同程度的抑菌作用,对化脓性链球菌、多杀性巴氏杆菌、金黄色葡萄球菌的抑菌作用最为明显,最小抑菌浓度均为49.5μg/mL,对猪链球菌和大肠杆菌也有较为明显的抑菌活性,而对猪支气管败血波氏杆菌的最小抑菌浓度为398μg/mL。当前在抗生素大量使用导致细菌产生广泛

  20. Pneumonia enzoótica em javalis (Sus scrofa Enzootic pneumonia in wild boars (Sus scrofa

    Directory of Open Access Journals (Sweden)

    Roselene Ecco

    2009-06-01

    Full Text Available O objetivo desse estudo foi descrever o quadro clínico e epidemiológico, os achados patológicos, bacteriológicos e imuno-histoquímicos de um surto de pneumonia em uma granja de Javalis do Distrito Federal, Brasil. Em um período de cinco meses, morreram 90 javalis. Desses, 63 tinham lesões pulmonares. Clinicamente apresentavam atraso no desenvolvimento corporal, diminuição do apetite, letargia, tosse e dificuldade respiratória, principalmente quando movimentados. Constatou-se elevação da temperatura, 40ºC em média. Na auscultação, havia crepitações e estertores pulmonares de intensidade moderada. As alterações macroscópicas nos pulmões analisados eram típicas de broncopneumonia lobular. As lesões caracterizavam-se por consolidação crânio-ventral na maioria dos pulmões. A coloração variava de difusamente vermelho-escuro a um padrão mosaico (lóbulos vermelho-escuros intercalados por lóbulos cinzas ou difusamente acinzentados. Na maioria dos pulmões observou-se exsudato mucopurulento na luz dos brônquios e fluindo do parênquima. Histologicamente, as alterações eram de broncopneumonia purulenta e histiocitária com focos de necrose. Em alguns animais havia também hiperplasia do BALT e, na maioria dos animais, infiltração linfocítica perivascular e peribronquial. Bordetella bronchiseptica e Streptococcus spp. foram as principais bactérias isoladas. A imuno-histoquímica demonstrou a bactéria Mycoplasma hyopneumoniae no epitélio bronquiolar e bronquial e o DNA desta bactéria foi detectado pela PCR. Este é o primeiro relato de broncopneumonia em Javalis associado à infecção por M. hyopneumoniae.The aim of this paper is to describe the clinical, epidemiological, pathological, bacteriological and immunohistochemical aspects of a pneumonia outbreak in a wild pig farm in the Distrito Federal, Brazil. Ninety wild pigs died in a period of five months, and 63 of these had pulmonary lesions. Clinically, the pigs

  1. Effects of aerosolized ketamine on the level of nitric oxide and nitric oxide synthetase in the lung tissue of rat with asthma

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Objective: To explore the effects of aerosolized ketamine on the level of nitric oxide and nitric oxide synthetase in the lung tissue in rat asthma model. Methods: Forty SD rats were randomly assigned to five groups: control group (group N), asthma model group (group A), two pretreated groups of different concentrations of ketamine (group K1, K2)and dexamethasone group(group D) with eight rats in each group. The rats in group A were sensitized by injection of ovalbumin (OA) together with aluminum hydroxide and bordetella pertussis as adjuvants. Two weeks after the sensitization, aerosolized OA was used to cause asthma. The rats in group K1 and K2 were sensitized with OA as group A , and then exposed to aerosol of ketamine , with the concentration of 25 g/L and 50 g/L respectively. Before using aerosolized OA, the rats in group D were exposed to aerosol of 0.01% dexamethasone . The level of NO2-/NO3- in lung tissues, inducible nitric oxide synthetase(iNOS) and constitute nitric oxide synthetase(cNOS) was measured in all groups. Results: The level of NO2-/NO3- and the activity of iNOS in lung tissues in group A were signiticantly higher than those in the other groups. The iNOS activity and the level of NO2-/NO3- in lung tissues were highly positively correlated. Conclusion: NO can induce airway hyperreactivity that may worsen asthma. Aerosolized ketamine can decrease the iNOS expression and reduce the level of NO in the lung tissue in rat asthma model.

  2. Structural and Functional Studies of WlbA: A Dehydrogenase Involved in the Biosynthesis of 2,3-Diacetamido-2,3-dideoxy-d-mannuronic Acid

    Energy Technology Data Exchange (ETDEWEB)

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

    2010-09-08

    2,3-Diacetamido-2,3-dideoxy-D-mannuronic acid (ManNAc3NAcA) is an unusual dideoxy sugar first identified nearly 30 years ago in the lipopolysaccharide of Pseudomonas aeruginosa O:3a,d. It has since been observed in other organisms, including Bordetella pertussis, the causative agent of whooping cough. Five enzymes are required for the biosynthesis of UDP-ManNAc3NAcA starting from UDP-N-acetyl-D-glucosamine. Here we describe a structural study of WlbA, the NAD-dependent dehydrogenase that catalyzes the second step in the pathway, namely, the oxidation of the C-3{prime} hydroxyl group on the UDP-linked sugar to a keto moiety and the reduction of NAD{sup +} to NADH. This enzyme has been shown to use {alpha}-ketoglutarate as an oxidant to regenerate the oxidized dinucleotide. For this investigation, three different crystal structures were determined: the enzyme with bound NAD(H), the enzyme in a complex with NAD(H) and {alpha}-ketoglutarate, and the enzyme in a complex with NAD(H) and its substrate (UDP-N-acetyl-D-glucosaminuronic acid). The tetrameric enzyme assumes an unusual quaternary structure with the dinucleotides positioned quite closely to one another. Both {alpha}-ketoglutarate and the UDP-linked sugar bind in the WlbA active site with their carbon atoms (C-2 and C-3{prime}, respectively) abutting the re face of the cofactor. They are positioned {approx}3 {angstrom} from the nicotinamide C-4. The UDP-linked sugar substrate adopts a highly unusual curved conformation when bound in the WlbA active site cleft. Lys 101 and His 185 most likely play key roles in catalysis.

  3. How long do nosocomial pathogens persist on inanimate surfaces? A systematic review

    Directory of Open Access Journals (Sweden)

    Schwebke Ingeborg

    2006-08-01

    Full Text Available Abstract Background Inanimate surfaces have often been described as the source for outbreaks of nosocomial infections. The aim of this review is to summarize data on the persistence of different nosocomial pathogens on inanimate surfaces. Methods The literature was systematically reviewed in MedLine without language restrictions. In addition, cited articles in a report were assessed and standard textbooks on the topic were reviewed. All reports with experimental evidence on the duration of persistence of a nosocomial pathogen on any type of surface were included. Results Most gram-positive bacteria, such as Enterococcus spp. (including VRE, Staphylococcus aureus (including MRSA, or Streptococcus pyogenes, survive for months on dry surfaces. Many gram-negative species, such as Acinetobacter spp., Escherichia coli, Klebsiella spp., Pseudomonas aeruginosa, Serratia marcescens, or Shigella spp., can also survive for months. A few others, such as Bordetella pertussis, Haemophilus influenzae, Proteus vulgaris, or Vibrio cholerae, however, persist only for days. Mycobacteria, including Mycobacterium tuberculosis, and spore-forming bacteria, including Clostridium difficile, can also survive for months on surfaces. Candida albicans as the most important nosocomial fungal pathogen can survive up to 4 months on surfaces. Persistence of other yeasts, such as Torulopsis glabrata, was described to be similar (5 months or shorter (Candida parapsilosis, 14 days. Most viruses from the respiratory tract, such as corona, coxsackie, influenza, SARS or rhino virus, can persist on surfaces for a few days. Viruses from the gastrointestinal tract, such as astrovirus, HAV, polio- or rota virus, persist for approximately 2 months. Blood-borne viruses, such as HBV or HIV, can persist for more than one week. Herpes viruses, such as CMV or HSV type 1 and 2, have been shown to persist from only a few hours up to 7 days. Conclusion The most common nosocomial pathogens may

  4. European Sero-Epidemiology Network 2: standardisation of immunoassay results for pertussis requires homogeneity in the antigenic preparations.

    Science.gov (United States)

    Giammanco, Anna; Nardone, Antony; Pebody, Richard; Kafatos, George; Andrews, Nick; Chiarini, Alfredo; Taormina, Susanna; de Ory, Fernando; Prosenc, Katarina; Krize, Bohumir; Hallander, Hans; Ljungman, Margaretha; Marva, Esther; Tsakris, Athanassios; O'Flanagan, Darina; Schneider, François; Griskevicius, Algirdas; Vranckx, Robert; Karacs, Ildiko

    2008-08-18

    A standardisation process, already developed during the earlier European Sero-Epidemiology Network (ESEN) project, was employed with a more robust algorithm to harmonise results of pertussis serological assays performed in 12 European and non-European countries. Initially, results from each country's own assay were compared with those obtained at the reference laboratory by means of an in-house pertussis toxin (PT)-based ELISA: seven countries used in-house or commercial PT-ELISAs; the other countries used assays based on Bordetella pertussis whole cell extracts (WCE) (three countries) or on combined PT-FHA (filamentous haemagglutinin) antigenic preparations (two countries). The WCE assays, although admitted for diagnostic purposes, confirmed their low correlation with the PT-ELISAs and their results could not be used for standardisation; the PT-FHA ELISAs gave results that were suitable for standardisation in one country but unsatisfactory in the other; the use of purified PT in serological assays confirmed its better reliability than other preparations and all PT-ELISAs results could be calibrated against those of the reference centre. In the standardisation process two high-titre cut-offs indicative of likelihood of recent infection (from within 4 weeks of disease onset up to 1 year after) were included for evaluations as they are suggested to be more useful, for the sero-epidemiological assays of immunity to pertussis, than the cut-off of protection, commonly employed, but still not defined for pertussis. Providing PT-ELISAs are used, standardisation of pertussis assay results is always possible and, when standardisation is performed, evaluation and comparison of the impact of different interventions can be also allowed, by measuring at the distribution of high antibody titres in the populations. PMID:18602434

  5. Altered serum microRNAs as biomarkers for the early diagnosis of pulmonary tuberculosis infection

    Directory of Open Access Journals (Sweden)

    Qi Yuhua

    2012-12-01

    Full Text Available Abstract Background Pulmonary tuberculosis (TB is a highly lethal infectious disease and early diagnosis of TB is critical for the control of disease progression. The objective of this study was to profile a panel of serum microRNAs (miRNAs as potential biomarkers for the early diagnosis of pulmonary TB infection. Methods Using TaqMan Low-Density Array (TLDA analysis followed by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR validation, expression levels of miRNAs in serum samples from 30 patients with active tuberculosis and 60 patients with Bordetella pertussis (BP, varicella-zoster virus (VZV and enterovirus (EV were analyzed. Results The Low-Density Array data showed that 97 miRNAs were differentially expressed in pulmonary TB patient sera compared with healthy controls (90 up-regulated and 7 down-regulated. Following qRT-PCR confirmation and receiver operational curve (ROC analysis, three miRNAs (miR-361-5p, miR-889 and miR-576-3p were shown to distinguish TB infected patients from healthy controls and other microbial infections with moderate sensitivity and specificity (area under curve (AUC value range, 0.711-0.848. Multiple logistic regression analysis of a combination of these three miRNAs showed an enhanced ability to discriminate between these two groups with an AUC value of 0.863. Conclusions Our study suggests that altered levels of serum miRNAs have great potential to serve as non-invasive biomarkers for early detection of pulmonary TB infection.

  6. Guanosine 5'-triphosphate binding protein (G/sub i/) and two additional pertussis toxin substrates associated with muscarinic receptors in rat heart myocytes: characterization and age dependency

    International Nuclear Information System (INIS)

    The coupling of muscarinic receptors with G-proteins was investigated in cultured myocytes prepared from the hearts of newborn rats. The coupling was investigated in both young (5 days after plating) and aged (14 days after plating) cultures, in view of the completely different effects of 5'-guanylyl imidodiphosphate [Gpp(NH)p] on muscarinic agonist binding to homogenates from young vs aged cultures. Pretreatment of cultures from both ages by Bordetella pertussis toxin (IAP) was found to eliminate any Gpp(NH)p effect on carbamylcholine binding. IAP by itself induced a rightward shift in the carbamylcholine competition curve in homogenates from aged cultures, but no such effect was observed in homogenates from young cultures. IAP-catalyzed [32P]ADP-ribosylation of membrane preparations from young and aged cultures revealed major differences between them. Young cultures exhibited a major IAP substrate at 40 kDa, which was also recognized by anti-α/sub i/ antibodies, and two novel IAP substrates at 28 and 42 kDa, which were weakly ADP-ribosylated by the toxin and were not recognized with either anti-α/sub i/ or anti-α0 antibodies. In aged cultures, only the 40-kDa band (ribosylated to a lower degree) was detected. The parallel age-dependent changes in the three IAP substrates (28, 40, and 42 kDa) and in the interactions of the G-protein(s) with the muscarinic receptors strongly suggest close association between the two phenomena. All of these age-dependent changes in the G-protein related parameters were prevented by phosphatidylcholine-liposome treatment of the aged cultures. The role of the membrane lipid composition in these phenomena is discussed

  7. In vivo evaluation of adeno-associated virus gene transfer in airways of mice with acute or chronic respiratory infection.

    Science.gov (United States)

    Myint, Melissa; Limberis, Maria P; Bell, Peter; Somanathan, Suryanarayan; Haczku, Angela; Wilson, James M; Diamond, Scott L

    2014-11-01

    Patients with cystic fibrosis (CF) often suffer chronic lung infection with concomitant inflammation, a setting that may reduce the efficacy of gene transfer. While gene therapy development for CF often involves viral-based vectors, little is known about gene transfer in the context of an infected airway. In this study, three mouse models were established to evaluate adeno-associated virus (AAV) gene transfer in such an environment. Bordetella bronchiseptica RB50 was used in a chronic, nonlethal respiratory infection in C57BL/6 mice. An inoculum of ∼10(5) CFU allowed B. bronchiseptica RB50 to persist in the upper and lower respiratory tracts for at least 21 days. In this infection model, administration of an AAV vector on day 2 resulted in 2.8-fold reduction of reporter gene expression compared with that observed in uninfected controls. Postponement of AAV administration to day 14 resulted in an even greater (eightfold) reduction of reporter gene expression, when compared with uninfected controls. In another infection model, Pseudomonas aeruginosa PAO1 was used to infect surfactant protein D (SP-D) or surfactant protein A (SP-A) knockout (KO) mice. With an inoculum of ∼10(5) CFU, infection persisted for 2 days in the nasal cavity of either mouse model. Reporter gene expression was approximately ∼2.5-fold lower compared with uninfected mice. In the SP-D KO model, postponement of AAV administration to day 9 postinfection resulted in only a two fold reduction in reporter gene expression, when compared with expression seen in uninfected controls. These results confirm that respiratory infections, both ongoing and recently resolved, decrease the efficacy of AAV-mediated gene transfer. PMID:25144316

  8. Susceptibility of canine and feline bacterial pathogens to pradofloxacin and comparison with other fluoroquinolones approved for companion animals.

    Science.gov (United States)

    Schink, Anne-Kathrin; Kadlec, Kristina; Hauschild, Tomasz; Brenner Michael, Geovana; Dörner, Julia C; Ludwig, Carolin; Werckenthin, Christiane; Hehnen, Hans-Robert; Stephan, Bernd; Schwarz, Stefan

    2013-02-22

    In this study, 908 bacterial pathogens from defined infections of dogs and cats were tested for their susceptibility to the novel fluoroquinolone pradofloxacin, which was approved in 2011 for use in cats and dogs. Most of the bacteria tested (Staphylococcus aureus, Staphylococcus pseudintermedius, Escherichia coli, β-haemolytic streptococci, Pasteurella multocida and Bordetella bronchiseptica) exhibited low pradofloxacin MIC(90) values of ≤ 0.25 μg/ml. Solely Proteus spp. and Pseudomonas aeruginosa had higher MIC(90) values of ≥ 4 μg/ml. Only six (3.4%) of 177 S. pseudintermedius and 12 (5.3%) of 227 E. coli isolates showed pradofloxacin MICs of ≥ 2 μg/ml. Analysis of the quinolone resistance determining regions of the target genes identified double mutations in GyrA that resulted in amino acid exchanges S83L+D87N or S83L+D87Y and single or double mutations in ParC that resulted in amino acid exchanges S80I or S80I+E84G in all 12 E. coli isolates. The six S. pseudintermedius isolates exhibited amino acid exchanges S84L or E88K in GyrA and S80I in GrlA. Comparative analysis of the MICs of pradofloxacin and the MICs determined for enrofloxacin and its main metabolite ciprofloxacin, but also marbofloxacin, orbifloxacin, difloxacin and ibafloxacin was conducted for the target pathogens S. pseudintermedius, E. coli and P. multocida. This comparison confirmed that pradofloxacin MICs were significantly lower than those of the other tested fluoroquinolones. PMID:22939523

  9. 鸡组织中硫酸粘杆菌素残留的微生物学检测方法研究%Determination of Colistin Sulphate Residues in Chicken Tissues by Microbiolgical Method

    Institute of Scientific and Technical Information of China (English)

    温芳; 肖希龙

    2006-01-01

    在国内首次以支气管炎博代特氏菌(Bordetella bronchiseptica)ATCC4617作为检测菌株,用PBS提取鸡肌肉组织,用5%硫酸提取鸡肝脏和肾脏组织中的硫酸粘杆菌素,应用一剂量法设计原理制备肌肉、肝脏和肾脏组织标准曲线.硫酸粘杆菌素检测限肌肉组织为0.15μg/g,肝脏和肾脏组织均为0.20μg/g,低于我国农业部和欧盟规定的粘杆菌素最高残留限量.肌肉、肝脏和肾脏组织标准曲线在0.025~6.4μg/mL范围内线性关系良好,r分别为0.997 0、0.999 4、0.999 0,n=9.以高、低(300mg/L、60 mg/L按效价计)两个剂量给鸡饮水给药,肌肉高、低剂量组均未检测出硫酸粘杆菌素残留,肝脏和肾脏低剂量组未检测出硫酸粘杆菌素残留,高剂量组0 h有微量残留.

  10. Lipid Requirements for the Enzymatic Activity of MraY Translocases and in Vitro Reconstitution of the Lipid II Synthesis Pathway.

    Science.gov (United States)

    Henrich, Erik; Ma, Yi; Engels, Ina; Münch, Daniela; Otten, Christian; Schneider, Tanja; Henrichfreise, Beate; Sahl, Hans-Georg; Dötsch, Volker; Bernhard, Frank

    2016-01-29

    Screening of new compounds directed against key protein targets must continually keep pace with emerging antibiotic resistances. Although periplasmic enzymes of bacterial cell wall biosynthesis have been among the first drug targets, compounds directed against the membrane-integrated catalysts are hardly available. A promising future target is the integral membrane protein MraY catalyzing the first membrane associated step within the cytoplasmic pathway of bacterial peptidoglycan biosynthesis. However, the expression of most MraY homologues in cellular expression systems is challenging and limits biochemical analysis. We report the efficient production of MraY homologues from various human pathogens by synthetic cell-free expression approaches and their subsequent characterization. MraY homologues originating from Bordetella pertussis, Helicobacter pylori, Chlamydia pneumoniae, Borrelia burgdorferi, and Escherichia coli as well as Bacillus subtilis were co-translationally solubilized using either detergent micelles or preformed nanodiscs assembled with defined membranes. All MraY enzymes originating from Gram-negative bacteria were sensitive to detergents and required nanodiscs containing negatively charged lipids for obtaining a stable and functionally folded conformation. In contrast, the Gram-positive B. subtilis MraY not only tolerates detergent but is also less specific for its lipid environment. The MraY·nanodisc complexes were able to reconstitute a complete in vitro lipid I and lipid II forming pipeline in combination with the cell-free expressed soluble enzymes MurA-F and with the membrane-associated protein MurG. As a proof of principle for future screening platforms, we demonstrate the inhibition of the in vitro lipid II biosynthesis with the specific inhibitors fosfomycin, feglymycin, and tunicamycin. PMID:26620564

  11. Structure of the N-terminal fragment of Escherichia coli Lon protease

    International Nuclear Information System (INIS)

    The medium-resolution structure of the N-terminal fragment of E. coli Lon protease shows that this part of the enzyme consists of two compact domains and a very long α-helix. The structure of a recombinant construct consisting of residues 1–245 of Escherichia coli Lon protease, the prototypical member of the A-type Lon family, is reported. This construct encompasses all or most of the N-terminal domain of the enzyme. The structure was solved by SeMet SAD to 2.6 Å resolution utilizing trigonal crystals that contained one molecule in the asymmetric unit. The molecule consists of two compact subdomains and a very long C-terminal α-helix. The structure of the first subdomain (residues 1–117), which consists mostly of β-strands, is similar to that of the shorter fragment previously expressed and crystallized, whereas the second subdomain is almost entirely helical. The fold and spatial relationship of the two subdomains, with the exception of the C-terminal helix, closely resemble the structure of BPP1347, a 203-amino-acid protein of unknown function from Bordetella parapertussis, and more distantly several other proteins. It was not possible to refine the structure to satisfactory convergence; however, since almost all of the Se atoms could be located on the basis of their anomalous scattering the correctness of the overall structure is not in question. The structure reported here was also compared with the structures of the putative substrate-binding domains of several proteins, showing topological similarities that should help in defining the binding sites used by Lon substrates

  12. Identification of Diversity-Generating Retroelements in Human Microbiomes

    Directory of Open Access Journals (Sweden)

    Yuzhen Ye

    2014-08-01

    Full Text Available Diversity-generating retroelements (DGRs are a unique family of retroelements that confer selective advantages to their hosts by accelerating the evolution of target genes through a specialized, error-prone, reverse transcription process. First identified in a Bordetella phage (BPP-1, which mediates the phage tropism specificity by generating variability in an involved gene, DGRs were predicted to be present in a larger collection of viral and bacterial species. A minimal DGR system is comprised of a reverse transcriptase (RTase gene, a template sequence (TR and a variable region (VR within a target gene. We developed a computational tool, DGRscan, to allow either de novo identification (based on the prediction of potential template-variable region pairs or similarity-based searches of DGR systems using known template sequences as the reference. The application of DGRscan to the human microbiome project (HMP datasets resulted in the identification of 271 non-redundant DGR systems, doubling the size of the collection of known DGR systems. We further identified a large number of putative target genes (651, which share no more than 90% sequence identity at the amino acid level that are potentially under diversification by the DGR systems. Our study provides the first survey of the DGR systems in the human microbiome, showing that the DGR systems are frequently found in human-associated bacterial communities, although they are of low incidence in individual genomes. Our study also provides functional clues for a large number of genes (reverse transcriptases and target genes that were previously annotated as proteins of unknown functions or nonspecific functions.

  13. The Campylobacter jejuni CiaC virulence protein is secreted from the flagellum and delivered to the cytosol of host cells

    Directory of Open Access Journals (Sweden)

    Jason eNeal-McKinney

    2012-03-01

    Full Text Available Campylobacter jejuni is a leading cause of bacterial gastroenteritis worldwide. Acute C. jejuni-mediated disease (campylobacteriosis involves C. jejuni invasion of host epithelial cells using a set of virulence proteins known as the Campylobacter invasion antigens (Cia. The genes encoding the Cia proteins are up-regulated upon co-culture of C. jejuni with epithelial cells. One of the Cia proteins, CiaC, is required for maximal invasion of host cells by C. jejuni. Previous work has also revealed that CiaC is, in part, responsible for host cell cytoskeletal rearrangements that result in membrane ruffling. This study was performed to test the hypothesis that CiaC is delivered to the cytosol of host cells. To detect the delivery of CiaC into cultured epithelial cells, we used the adenylate cyclase domain (ACD of Bordetella pertussis CyaA as a reporter. In this study, we found that export and delivery of the C. jejuni Cia proteins into human INT 407 epithelial cells required a functional flagellar hook complex composed of FlgE, FlgK, and FlgL. Assays performed with bacterial culture supernatants supported the hypothesis that CiaC delivery requires bacteria-host cell contact. We also found that that CiaC was delivered to host cells by cell-associated (bound bacteria, as judged by experiments performed with inhibitors that specifically target the cell signaling pathways utilized by C. jejuni for cell invasion. Interestingly, the C. jejuni flgL mutant, which is incapable of exporting and delivering the Cia proteins, did not induce INT 407 cell membrane ruffles. Complementation of the flgL mutant with plasmid-encoded flgL restored the motility and membrane ruffling. These data support the hypothesis that the C. jejuni Cia proteins, which are exported from the flagellum, are delivered to the cytosol of host cells.

  14. Calcium influx rescues adenylate cyclase-hemolysin from rapid cell membrane removal and enables phagocyte permeabilization by toxin pores.

    Directory of Open Access Journals (Sweden)

    Radovan Fiser

    Full Text Available Bordetella adenylate cyclase toxin-hemolysin (CyaA penetrates the cytoplasmic membrane of phagocytes and employs two distinct conformers to exert its multiple activities. One conformer forms cation-selective pores that permeabilize phagocyte membrane for efflux of cytosolic potassium. The other conformer conducts extracellular calcium ions across cytoplasmic membrane of cells, relocates into lipid rafts, translocates the adenylate cyclase enzyme (AC domain into cells and converts cytosolic ATP to cAMP. We show that the calcium-conducting activity of CyaA controls the path and kinetics of endocytic removal of toxin pores from phagocyte membrane. The enzymatically inactive but calcium-conducting CyaA-AC⁻ toxoid was endocytosed via a clathrin-dependent pathway. In contrast, a doubly mutated (E570K+E581P toxoid, unable to conduct Ca²⁺ into cells, was rapidly internalized by membrane macropinocytosis, unless rescued by Ca²⁺ influx promoted in trans by ionomycin or intact toxoid. Moreover, a fully pore-forming CyaA-ΔAC hemolysin failed to permeabilize phagocytes, unless endocytic removal of its pores from cell membrane was decelerated through Ca²⁺ influx promoted by molecules locked in a Ca²⁺-conducting conformation by the 3D1 antibody. Inhibition of endocytosis also enabled the native B. pertussis-produced CyaA to induce lysis of J774A.1 macrophages at concentrations starting from 100 ng/ml. Hence, by mediating calcium influx into cells, the translocating conformer of CyaA controls the removal of bystander toxin pores from phagocyte membrane. This triggers a positive feedback loop of exacerbated cell permeabilization, where the efflux of cellular potassium yields further decreased toxin pore removal from cell membrane and this further enhances cell permeabilization and potassium efflux.

  15. A Study on Co-Metabolic Biodegradation of Phenol and TCE by Mixed Culture PCL%高效降解菌剂PCL对苯酚及TCE的共代谢降解性能研究

    Institute of Scientific and Technical Information of China (English)

    张施阳

    2016-01-01

    从长期受氯代烃和酚类污染的土壤中驯化筛选得到能以苯酚为共代谢基质降解TCE的混合菌群PCL,其主要由博代氏杆菌(Bordetella sp.)和中华根瘤菌(Sinorhizobium sp.)组成.采用摇瓶振荡培养方法,研究了不同环境条件对混合菌群PCL共代谢苯酚和TCE效能的影响.结果表明,在30℃、pH 7.5条件下,200 mg/L苯酚能被PCL在4d内代谢完全,并且50 mg/L TCE在8d内的降解率能达到85.6%,以8mmol/L H2O2作为氧源,能使TCE降解率提高到91.18%.混合菌群PCL对苯酚和TCE的降解动力学可用Haldane方程来描述,利用非线性拟合求得各自对应的动力学参数k、km、ki分别为0.749/d、46.67 mg/L、386.16 mg/L和0.1332/d、31.15mg/L、0.044 5 mg/L,且拟合效果很好.

  16. Microbes bind complement inhibitor factor H via a common site.

    Science.gov (United States)

    Meri, T; Amdahl, H; Lehtinen, M J; Hyvärinen, S; McDowell, J V; Bhattacharjee, A; Meri, S; Marconi, R; Goldman, A; Jokiranta, T S

    2013-01-01

    To cause infections microbes need to evade host defense systems, one of these being the evolutionarily old and important arm of innate immunity, the alternative pathway of complement. It can attack all kinds of targets and is tightly controlled in plasma and on host cells by plasma complement regulator factor H (FH). FH binds simultaneously to host cell surface structures such as heparin or glycosaminoglycans via domain 20 and to the main complement opsonin C3b via domain 19. Many pathogenic microbes protect themselves from complement by recruiting host FH. We analyzed how and why different microbes bind FH via domains 19-20 (FH19-20). We used a selection of FH19-20 point mutants to reveal the binding sites of several microbial proteins and whole microbes (Haemophilus influenzae, Bordetella pertussis, Pseudomonas aeruginosa, Streptococcus pneumonia, Candida albicans, Borrelia burgdorferi, and Borrelia hermsii). We show that all studied microbes use the same binding region located on one side of domain 20. Binding of FH to the microbial proteins was inhibited with heparin showing that the common microbial binding site overlaps with the heparin site needed for efficient binding of FH to host cells. Surprisingly, the microbial proteins enhanced binding of FH19-20 to C3b and down-regulation of complement activation. We show that this is caused by formation of a tripartite complex between the microbial protein, FH, and C3b. In this study we reveal that seven microbes representing different phyla utilize a common binding site on the domain 20 of FH for complement evasion. Binding via this site not only mimics the glycosaminoglycans of the host cells, but also enhances function of FH on the microbial surfaces via the novel mechanism of tripartite complex formation. This is a unique example of convergent evolution resulting in enhanced immune evasion of important pathogens via utilization of a "superevasion site." PMID:23637600

  17. Assembly and function of type III secretory systems.

    Science.gov (United States)

    Cornelis, G R; Van Gijsegem, F

    2000-01-01

    Type III secretion systems allow Yersinia spp., Salmonella spp., Shigella spp., Bordetella spp., and Pseudomonas aeruginosa and enteropathogenic Escherichia coli adhering at the surface of a eukaryotic cell to inject bacterial proteins across the two bacterial membranes and the eukaryotic cell membrane to destroy or subvert the target cell. These systems consist of a secretion apparatus, made of approximately 25 proteins, and an array of proteins released by this apparatus. Some of these released proteins are "effectors," which are delivered into the cytosol of the target cell, whereas the others are "translocators," which help the effectors to cross the membrane of the eukaryotic cell. Most of the effectors act on the cytoskeleton or on intracellular-signaling cascades. A protein injected by the enteropathogenic E. coli serves as a membrane receptor for the docking of the bacterium itself at the surface of the cell. Type III secretion systems also occur in plant pathogens where they are involved both in causing disease in susceptible hosts and in eliciting the so-called hypersensitive response in resistant or nonhost plants. They consist of 15-20 Hrp proteins building a secretion apparatus and two groups of effectors: harpins and avirulence proteins. Harpins are presumably secreted in the extracellular compartment, whereas avirulence proteins are thought to be targeted into plant cells. Although a coherent picture is clearly emerging, basic questions remain to be answered. In particular, little is known about how the type III apparatus fits together to deliver proteins in animal cells. It is even more mysterious for plant cells where a thick wall has to be crossed. In spite of these haunting questions, type III secretion appears as a fascinating trans-kingdom communication device. PMID:11018143

  18. Aerosolized Bacillus anthracis infection in New Zealand white rabbits: natural history and intravenous levofloxacin treatment.

    Science.gov (United States)

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

    2010-12-01

    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 LD(50) 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 preceded both pyrexia and inversion of the heterophil:lymphocyte ratio, an indicator of infection. Antigenemia was determined within 1 h by electrochemiluminescence immunoassay, compared with the 24-h traditional culture needed for bacteremia determination. Rabbits appeared clinically normal until shortly before succumbing to anthrax approximately 47 h after challenge or approximately 22 h after antigenemia, which suggests a relatively narrow therapeutic window of opportunity. To evaluate the therapeutic rabbit model, B. anthracis-exposed rabbits were treated (after determination of antigenemia and later confirmed to be bacteremic) intravenously with the fluoroquinolone antibiotic levofloxacin for 5 d at a total daily dose of 25 or 12.5 mg/kg, resulting in nearly 90% and 70% survival, respectively, to the study end (28 d after challenge). The peak level for 12.5 mg/kg was equivalent to that observed for a 500-mg daily levofloxacin dose in humans. These results suggest that intravenous levofloxacin is an effective therapeutic against inhalational anthrax. Taken together, our findings indicate that antigenemia is a viable and early biomarker for B. anthracis infection that can be used as a treatment trigger to allow for timely intervention against this highly pathogenic disease. PMID:21262133

  19. Avaliação da contaminação microbiana e de parâmetros ambientais – temperatura, ventilação e umidade - na Biblioteca Central da Universidade Federal de Santa Maria/RS: acervos da Coleção Teses e Coletânea UFSM

    Directory of Open Access Journals (Sweden)

    Lizandra Veleda Arabidian

    2014-12-01

    Full Text Available Este artigo analisará um estudo da biodeterioração em dois acervos que fazem parte do patrimônio documental da UFSM. O estudo enfatiza a importância da preservação documental, uma das funções fundamentais para a conservação dos acervos de bibliotecas. A pesquisa tem foco nos problemas ambientais avaliados, que são os microrganismos presentes no ambiente e as condições de umidade, iluminação, ventilação e temperatura no período de um ano. O diagnóstico foi realizado através da observação direta dos acervos, permitindo detectar as necessidades para conservação e preservação destes ambientes através de um plano de ações. Para a realização deste trabalho, foram estudados referenciais teóricos, levantamentos fotográficos, assim como a coleta e identificação de agentes microbiológicos, das condições de umidade, iluminação, ventilação e temperatura no ambiente. Os resultados identificaram as bactérias Bacillus sp., Serratia plymuthica, Acinetobacter sp., Bordetella trematum, Pseudomonas sp., Enterobacter agglomerans, Moraxella sp., Micrococcus, Staphylococcus coagulase negativa, Staphylococcus epidermidis, Aeromonas sp., Enterococcus sp., Cedecea lapagei, Nocardia sp., Pseudomonas aeruginosa e fungos Penicillium sp., Fusarium sp.e Cladosporium sp., considerados microrganismos patogênicos ou alergênicos. Na análise dos dados verificou-se que há a necessidade de usar recursos para a correção de índices elevados de umidade e temperatura no ambiente da Biblioteca da UFSM.

  20. Protein chainmail variants in dsDNA viruses

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    Z. Hong Zhou

    2015-06-01

    Full Text Available First discovered in bacteriophage HK97, biological chainmail is a highly stable system formed by concatenated protein rings. Each subunit of the ring contains the HK97-like fold, which is characterized by its submarine-like shape with a 5-stranded β sheet in the axial (A domain, spine helix in the peripheral (P domain, and an extended (E loop. HK97 capsid consists of covalently-linked copies of just one HK97-like fold protein and represents the most effective strategy to form highly stable chainmail needed for dsDNA genome encapsidation. Recently, near-atomic resolution structures enabled by cryo electron microscopy (cryoEM have revealed a range of other, more complex variants of this strategy for constructing dsDNA viruses. The first strategy, exemplified by P22-like phages, is the attachment of an insertional (I domain to the core 5-stranded β sheet of the HK97-like fold. The atomic models of the Bordetella phage BPP-1 showcases an alternative topology of the classic HK97 topology of the HK97-like fold, as well as the second strategy for constructing stable capsids, where an auxiliary jellyroll protein dimer serves to cement the non-covalent chainmail formed by capsid protein subunits. The third strategy, found in lambda-like phages, uses auxiliary protein trimers to stabilize the underlying non-covalent chainmail near the 3-fold axis. Herpesviruses represent highly complex viruses that use a combination of these strategies, resulting in four-level hierarchical organization including a non-covalent chainmail formed by the HK97-like fold domain found in the floor region. A thorough understanding of these structures should help unlock the enigma of the emergence and evolution of dsDNA viruses and inform bioengineering efforts based on these viruses.

  1. Rediscovering Pertussis

    Science.gov (United States)

    Zlamy, Manuela

    2016-01-01

    Pertussis, caused by Bordetella (B.) pertussis, a Gram-negative bacterium, is a highly contagious airway infection. Especially in infants, pertussis remains a major health concern. Acute infection with B. pertussis can cause severe illness characterized by severe respiratory failure, pulmonary hypertension, leucocytosis, and death. Over the past years, rising incidence rates of intensive care treatment in young infants were described. Due to several virulence factors (pertussis toxin, tracheal cytotoxin, adenylate cyclase toxin, filamentous hemagglutinin, and lipooligosaccharide) that promote bacterial adhesion and invasion, B. pertussis creates a unique niche for colonization within the human respiratory tract. The resulting long-term infection is mainly caused by the ability of B. pertussis to interfere with the host’s innate and adaptive immune system. Although pertussis is a vaccine-preventable disease, it has persisted in vaccinated populations. Epidemiological data reported a worldwide increase in pertussis incidence among children during the past years. Either acellular pertussis (aP) vaccines or whole-cell vaccines are worldwide used. Recent studies did not detect any differences according to pertussis incidence when comparing the different vaccines used. Most of the currently used aP vaccines protect against acute infections for a period of 6–8 years. The resurgence of pertussis may be due to the lack of herd immunity caused by missing booster immunizations among adolescents and adults, low vaccine coverages in some geographic areas, and genetic changes of different B. pertussis strains. Due to the rising incidence of pertussis, probable solution strategies are discussed. Cocooning strategies (vaccination of close contact persons) and immunizations during pregnancy appear to be an approach to reduce neonatal contagiousness. During the past years, studies focused on the pathway of the immune modulation done by B. pertussis to provide a basis for the

  2. Sequence-Based Screening for Rare Enzymes: New Insights into the World of AMDases Reveal a Conserved Motif and 58 Novel Enzymes Clustering in Eight Distinct Families.

    Science.gov (United States)

    Maimanakos, Janine; Chow, Jennifer; Gaßmeyer, Sarah K; Güllert, Simon; Busch, Florian; Kourist, Robert; Streit, Wolfgang R

    2016-01-01

    Arylmalonate Decarboxylases (AMDases, EC 4.1.1.76) are very rare and mostly underexplored enzymes. Currently only four known and biochemically characterized representatives exist. However, their ability to decarboxylate α-disubstituted malonic acid derivatives to optically pure products without cofactors makes them attractive and promising candidates for the use as biocatalysts in industrial processes. Until now, AMDases could not be separated from other members of the aspartate/glutamate racemase superfamily based on their gene sequences. Within this work, a search algorithm was developed that enables a reliable prediction of AMDase activity for potential candidates. Based on specific sequence patterns and screening methods 58 novel AMDase candidate genes could be identified in this work. Thereby, AMDases with the conserved sequence pattern of Bordetella bronchiseptica's prototype appeared to be limited to the classes of Alpha-, Beta-, and Gamma-proteobacteria. Amino acid homologies and comparison of gene surrounding sequences enabled the classification of eight enzyme clusters. Particularly striking is the accumulation of genes coding for different transporters of the tripartite tricarboxylate transporters family, TRAP transporters and ABC transporters as well as genes coding for mandelate racemases/muconate lactonizing enzymes that might be involved in substrate uptake or degradation of AMDase products. Further, three novel AMDases were characterized which showed a high enantiomeric excess (>99%) of the (R)-enantiomer of flurbiprofen. These are the recombinant AmdA and AmdV from Variovorax sp. strains HH01 and HH02, originated from soil, and AmdP from Polymorphum gilvum found by a data base search. Altogether our findings give new insights into the class of AMDases and reveal many previously unknown enzyme candidates with high potential for bioindustrial processes. PMID:27610105

  3. Proteomic analysis of growth phase-dependent expression of Legionella pneumophila proteins which involves regulation of bacterial virulence traits.

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    Tsuyoshi Hayashi

    Full Text Available Legionella pneumophila, which is a causative pathogen of Legionnaires' disease, expresses its virulent traits in response to growth conditions. In particular, it is known to become virulent at a post-exponential phase in vitro culture. In this study, we performed a proteomic analysis of differences in expression between the exponential phase and post-exponential phase to identify candidates associated with L. pneumophila virulence using 2-Dimentional Fluorescence Difference Gel Electrophoresis (2D-DIGE combined with Matrix-Assisted Laser Desorption/Ionization-Mass Spectrometry (MALDI-TOF-MS. Of 68 identified proteins that significantly differed in expression between the two growth phases, 64 were up-regulated at a post-exponential phase. The up-regulated proteins included enzymes related to glycolysis, ketone body biogenesis and poly-3-hydroxybutyrate (PHB biogenesis, suggesting that L. pneumophila may utilize sugars and lipids as energy sources, when amino acids become scarce. Proteins related to motility (flagella components and twitching motility-associated proteins were also up-regulated, predicting that they enhance infectivity of the bacteria in host cells under certain conditions. Furthermore, 9 up-regulated proteins of unknown function were found. Two of them were identified as novel bacterial factors associated with hemolysis of sheep red blood cells (SRBCs. Another 2 were found to be translocated into macrophages via the Icm/Dot type IV secretion apparatus as effector candidates in a reporter assay with Bordetella pertussis adenylate cyclase. The study will be helpful for virulent analysis of L. pneumophila from the viewpoint of physiological or metabolic modulation dependent on growth phase.

  4. Rediscovering Pertussis.

    Science.gov (United States)

    Zlamy, Manuela

    2016-01-01

    Pertussis, caused by Bordetella (B.) pertussis, a Gram-negative bacterium, is a highly contagious airway infection. Especially in infants, pertussis remains a major health concern. Acute infection with B. pertussis can cause severe illness characterized by severe respiratory failure, pulmonary hypertension, leucocytosis, and death. Over the past years, rising incidence rates of intensive care treatment in young infants were described. Due to several virulence factors (pertussis toxin, tracheal cytotoxin, adenylate cyclase toxin, filamentous hemagglutinin, and lipooligosaccharide) that promote bacterial adhesion and invasion, B. pertussis creates a unique niche for colonization within the human respiratory tract. The resulting long-term infection is mainly caused by the ability of B. pertussis to interfere with the host's innate and adaptive immune system. Although pertussis is a vaccine-preventable disease, it has persisted in vaccinated populations. Epidemiological data reported a worldwide increase in pertussis incidence among children during the past years. Either acellular pertussis (aP) vaccines or whole-cell vaccines are worldwide used. Recent studies did not detect any differences according to pertussis incidence when comparing the different vaccines used. Most of the currently used aP vaccines protect against acute infections for a period of 6-8 years. The resurgence of pertussis may be due to the lack of herd immunity caused by missing booster immunizations among adolescents and adults, low vaccine coverages in some geographic areas, and genetic changes of different B. pertussis strains. Due to the rising incidence of pertussis, probable solution strategies are discussed. Cocooning strategies (vaccination of close contact persons) and immunizations during pregnancy appear to be an approach to reduce neonatal contagiousness. During the past years, studies focused on the pathway of the immune modulation done by B. pertussis to provide a basis for the

  5. Protecting Newborns Against Pertussis: Treatment and Prevention Strategies.

    Science.gov (United States)

    Salim, Abdulbaset M; Liang, Yan; Kilgore, Paul E

    2015-12-01

    Pertussis is a potentially severe respiratory disease, which affects all age groups from young infants to older adults and is responsible for an estimated 195,000 deaths occurred globally in 2008. Active research is ongoing to better understand the pathogenesis, immunology, and diagnosis of pertussis. For diagnosis, molecular assays (e.g., polymerase chain reaction) for detection of Bordetella pertussis have become more widely available and support improved outbreak detection. In children, pertussis vaccines have been incorporated into routine immunization schedules and deployed for pertussis outbreak control. Lower levels of vaccine coverage are now being observed in communities where vaccine hesitancy is rising. Additionally, recognition that newborn babies are at risk of pertussis in the USA and UK has led to recommendations to immunize pregnant women. Among adolescents and older adults in the USA, Tetanus Toxoid, Reduced Diphtheria Toxoid and Acellular pertussis (Tdap) Vaccines are recommended, but substantial individual- and system-level barriers exist that will make achieving national Healthy People 2020 targets for immunization challenging. Current antimicrobial regimens for pertussis are focused on reducing the severity of disease, reducing rates of sequelae, and minimizing transmission of infection to susceptible individuals. Continued surveillance for pertussis will be important to identify opportunities for reducing young infants' exposure and reducing the impact of outbreaks among school-aged children. Laboratory-based surveillance for newly emerging strains of B. pertussis will be important to identify strains that may evade protection elicited by currently available vaccines. Efforts to develop new-generation pertussis vaccines should be considered now in anticipation of vaccine development programs, which may require ten or more years to deliver a licensed vaccine. PMID:26542059

  6. La vacunación contra pertussis: Estado actual y perspectivas futuras

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    Celso Pérez-Bolaños

    2012-01-01

    Full Text Available La tosferina o pertussis es una enfermedad producida por la bacteria Gram negativa Bordetella pertussis y constituye actualmente un problema de salud a escala mundial, siendo la población infantil la más vulnerable dentro de los grupos de riesgo. Suele ser fatal en niños menores de un año, debido a que la tos paroxística característica bloquea las vías respiratorias, lo cual provoca la asfixia y la muerte. Adicionalmente, se observan eventos adversos o complicaciones serias de la enfermedad tales como neumonías bacterianas secundarias, epilepsias, encefalopatía, apnea e hipertensión pulmonar. En la actualidad se aplican programas extensivos de vacunación contra pertussis en todo el mundo, de modo que la enfermedad está considerada como relativamente bien controlada. Según informes de la Organización Mundial de la Salud, cerca del 82 % de toda la población infantil mundial recibió las tres dosis de la vacuna de pertussis en 2008. La vacunación global contra pertussis en ese año evitó cerca de 687 000 muertes por la enfermedad. Sin embargo, se ha observado que ocurre un resurgimiento de ella a escala mundial, así como una prevalencia en zonas del mundo donde la vacunación es pobre. El presente trabajo tuvo como objetivo presentar una revisión acerca de los problemas inherentes a la vacunación contra esta enfermedad en el contexto actual de su resurgimiento a escala universal, la forma de enfocarlos y resolverlos, para su aplicación en Cuba en caso necesario. La vacunación contra pertussis necesita de nuevos enfoques dirigidos a la eliminación de un flagelo que afecta primordialmente a la población infantil, que es el futuro de la humanidad. La solución del problema parece ser la búsqueda de nuevas vacunas más eficaces y la extensión de los esquemas de vacunación a la población de adolescentes y adultos que rodean a los infantes, así como a las mujeres en estado de gestación y a los recién nacidos.

  7. Immunization: a key to primary health care.

    Science.gov (United States)

    1983-01-01

    Focus of this discussion is on some of the problems enountered by national immunization programs and on the technology that is available now or that will be in the near future to help solve these problems. 4 basic aspects of immunization services are examined: the safety, effectiveness, and stability of vaccines; the cold chain, i.e., the transportation, storage, and handling of heat-sensitives vaccines from manufacturer to health worker in the field; vaccination equipment and sterilization for correct administration of immunization; and program management--schedules, records, training, resource allocation. The section devoted to vaccines focuses on immunization against 6 of (diphtheria, whooping cough, tetanus, measles, polio, and tuberculosis) against 6 of the major killers of children in developing countries: BCG, DPT (diphtheria-pertussis-tetanus), measles and poli vaccines, and tetanus toxiod. The bacillus of Calmette and Guerline (BCG) is considered a very safe vaccine. Questions about the effectiveness of BCG in preventing tuberculosis have been raised throughout its 60-year history. Different studies have produced conflicting results, some showing BCG to be highly effective and others showing no positive effect. Diphtheria toxioid, a very safe and relatively stable vaccine, is very effective in protecting against the development of diphtheria. Live attenuated measles virus vaccine is a safe, highly effective vaccine, but it requires careful handling and storage to prevent damage due to excessive heat or light exposure. The vaccine used for pertussis (whooping cough) is a saline suspension of killed Bordetella pertussis bacteria. The vaccine usually is administered as part of the triple DPT vaccine. Concerns about its safety have led to greatly reduced levels of use in some European countries in recent years. Its effectiveness also has been questioned. 2 types of polio vaccine are available: a live, attenuated vaccine given orally (Sabin) and a killed or

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

    Energy Technology Data Exchange (ETDEWEB)

    Cummings, J.; Fedorov, A; Xu, C; Brown, S; Fedorov, E; Babbitt, P; Almo, S; Raushel, F

    2009-01-01

    The catalytic activities of three members of the amidohydrolase superfamily were discovered using amino acid substrate libraries. Bb3285 from Bordetella bronchiseptica, Gox1177 from Gluconobacter oxidans, 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, N-succinyl-d-Xaa, and l-Xaa-d-Xaa were used to establish the substrate profiles for these enzymes. It was demonstrated that Bb3285 is restricted to the hydrolysis of N-acyl-substituted derivatives of d-glutamate. The best substrates for this enzyme are N-formyl-d-glutamate (k{sub cat}/K{sub m} = 5.8 x 10{sup 6} M{sup -1} s{sup -1}), N-acetyl-d-glutamate (k{sub cat}/K{sub m} = 5.2 x 10{sup 6} M{sup -1} s{sup -1}), and l-methionine-d-glutamate (k{sub cat}/K{sub m} = 3.4 x 10{sup 5} M{sup -1} s{sup -1}). Gox1177 and Sco4986 preferentially hydrolyze N-acyl-substituted derivatives of hydrophobic d-amino acids. The best substrates for Gox1177 are N-acetyl-d-leucine (k{sub cat}/K{sub m} = 3.2 x 104 M{sup -1} s-1), N-acetyl-d-tryptophan (kcat/Km = 4.1 x 104 M-1 s-1), and l-tyrosine-d-leucine (kcat/Km = 1.5 x 104 M-1 s-1). A fourth protein, Bb2785 from B. bronchiseptica, did not have d-aminoacylase activity. The best substrates for Sco4986 are N-acetyl-d-phenylalanine and N-acetyl-d-tryptophan. The three-dimensional structures of Bb3285 in the presence of the product acetate or a potent mimic of the tetrahedral intermediate were determined by X-ray diffraction methods. The side chain of the d-glutamate moiety of the inhibitor is ion-paired to Arg-295, while the {alpha}-carboxylate is ion-paired with Lys-250 and Arg-376. These results have revealed the chemical and structural determinants for substrate specificity in this protein. Bioinformatic analyses of an additional {approx}250

  9. Annotating enzymes of uncertain function: the deacylation of D-amino acids by members of the amidohydrolase superfamily.

    Science.gov (United States)

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

    2009-07-14

    The catalytic activities of three members of the amidohydrolase superfamily were discovered using amino acid substrate libraries. Bb3285 from Bordetella bronchiseptica, Gox1177 from Gluconobacter oxidans, 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, N-succinyl-d-Xaa, and l-Xaa-d-Xaa were used to establish the substrate profiles for these enzymes. It was demonstrated that Bb3285 is restricted to the hydrolysis of N-acyl-substituted derivatives of d-glutamate. The best substrates for this enzyme are N-formyl-d-glutamate (k(cat)/K(m) = 5.8 x 10(6) M(-1) s(-1)), N-acetyl-d-glutamate (k(cat)/K(m) = 5.2 x 10(6) M(-1) s(-1)), and l-methionine-d-glutamate (k(cat)/K(m) = 3.4 x 10(5) M(-1) s(-1)). Gox1177 and Sco4986 preferentially hydrolyze N-acyl-substituted derivatives of hydrophobic d-amino acids. The best substrates for Gox1177 are N-acetyl-d-leucine (k(cat)/K(m) = 3.2 x 10(4) M(-1) s(-1)), N-acetyl-d-tryptophan (k(cat)/K(m) = 4.1 x 10(4) M(-1) s(-1)), and l-tyrosine-d-leucine (k(cat)/K(m) = 1.5 x 10(4) M(-1) s(-1)). A fourth protein, Bb2785 from B. bronchiseptica, did not have d-aminoacylase activity. The best substrates for Sco4986 are N-acetyl-d-phenylalanine and N-acetyl-d-tryptophan. The three-dimensional structures of Bb3285 in the presence of the product acetate or a potent mimic of the tetrahedral intermediate were determined by X-ray diffraction methods. The side chain of the d-glutamate moiety of the inhibitor is ion-paired to Arg-295, while the alpha-carboxylate is ion-paired with Lys-250 and Arg-376. These results have revealed the chemical and structural determinants for substrate specificity in this protein. Bioinformatic analyses of an additional approximately 250 sequences identified as members of this group

  10. Memory cell generation ablated by soluble protein antigen by means of effects on T- and B-lymphocyte compartments.

    Science.gov (United States)

    Karvelas, M; Nossal, G J

    1992-04-01

    Adult C57BL/6 mice were injected with 100 micrograms of soluble, freshly deaggregated human serum albumin (HSA) to produce partial immunologic tolerance. Uninjected normal control (N) mice contain only approximately 100 B cells in their spleens with the capacity to (i) be activated in vitro into clonal proliferation by Escherichia coli lipopolysaccharide plus interleukins 2, 4, and 5, (ii) form IgG1 as well as IgM antibody, and (iii) display specificity for HSA when only IgG1 is allowed to score in an enzyme-linked immunosorbent assay (ELISA). Such N mice generate approximately 50,000 clonable anti-HSA IgG1 antibody-forming cell precursors in their spleens after T-dependent immunization with HSA absorbed onto alum and given with Bordetella pertussis adjuvant. Mice preinjected with soluble HSA (TOL) generate far fewer anti-HSA IgG1 antibody-forming cell precursors, termed anti-HSA memory cells. Splenocytes were transferred from N or TOL mice into lethally irradiated syngeneic recipients together with syngeneic bone marrow. Whereas N splenocytes generated plentiful memory cells within 2 weeks in antigenically challenged recipients, TOL splenocytes did not. Work with Ly-5 congenic mice ruled out memory cell generation from either the host or the bone marrow inoculum within this limited time. N T cells plus TOL B cells showed consistently lowered memory cell generation. TOL T cells plus N B cells showed an even greater lowering of adoptive memory cell generation. Thus the lowered response capacity of TOL mice resided in the T- and B-cell compartments. Attempts to show a suppressor component within the T-cell population were inconclusive, but a profound defect in capacity to respond to HSA in vitro was exhibited by the CD4+ T cells of TOL mice. B lymphocytes were harvested from T-dependently immunized mice 5 days after challenge, incubated with soluble HSA for 18 hr, and then adoptively transferred together with N T cells. The recently activated B cells were not

  11. Desarrollo de vacunas antipertúsicas via genómica-proteómica

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    Celso Pérez-Bolaños

    2013-01-01

    Full Text Available Las vacunas convencionales se basan en la aplicación de diversos métodos bioquímicos, microbiológicos e inmunológicos sobre los agentes causales de las enfermedades. A pesar del éxito alcanzado en el control y prevención vacunal de muchas patologías infecciosas, en otros casos los resultados no han sido fructíferos, de modo que se impone explorar nuevos enfoques y posibilidades. El desarrollo y auge de la biología molecular representa una revolución en el diseño de nuevas vacunas contra una gran variedad de microorganismos patógenos, causantes de enfermedades que afectan al hombre y a los animales. A partir del conocimiento de las secuencias genómicas completas de esos microorganismos, en combinación con nuevas tecnologías avanzadas tales como la bioinformática, los micromatrices de ADN y la proteómica, ha sido posible identificar nuevos antígenos importantes para el desarrollo de nuevos preparados vacunales. La combinación genómica-proteómica ha reportado grandes avances en el descubrimiento de sitios "diana" de vacunas contra bacterias patógenas. La vacuna ideal debe ser patógeno - específica y portadora de antígenos conservados en todas las cepas, de modo que posea una cobertura universal, con el mínimo número de antígenos. Apoyándose en estos conocimientos es posible predecir a priori todos los antígenos in silico, sin necesidad de crecer el microorganismo in vitro. Este nuevo enfoque ha sido aplicado con éxito en la identificación de nuevos antígenos que pueden ser evaluados de manera perspectiva para fines vacunales en Bordetella pertussis, lo que evidentemente representa un serio esfuerzo a escala global para enfrentar la resurgencia de la tosferina o pertussis.

  12. Vaccine-Mediated Activation of Human TLR4 Is Affected by Modulation of Culture Conditions during Whole-Cell Pertussis Vaccine Preparation.

    Science.gov (United States)

    Hoonakker, Marieke E; Verhagen, Lisa M; Pupo, Elder; de Haan, Alex; Metz, Bernard; Hendriksen, Coenraad F M; Han, Wanda G H; Sloots, Arjen

    2016-01-01

    The potency of whole-cell pertussis (wP) vaccines is still determined by an intracerebral mouse protection test. To allow development of suitable in vitro alternatives to this test, insight into relevant parameters to monitor the consistency of vaccine quality is essential. To this end, a panel of experimental wP vaccines of varying quality was prepared by sulfate-mediated suppression of the BvgASR master virulence regulatory system of Bordetella pertussis during cultivation. This system regulates the transcription of a range of virulence proteins, many of which are considered important for the induction of effective host immunity. The protein compositions and in vivo potencies of the vaccines were BvgASR dependent, with the vaccine containing the highest amount of virulence proteins having the highest in vivo potency. Here, the capacities of these vaccines to stimulate human Toll-like receptors (hTLR) 2 and 4 and the role these receptors play in wP vaccine-mediated activation of antigen-presenting cells in vitro were studied. Prolonged BvgASR suppression was associated with a decreased capacity of vaccines to activate hTLR4. In contrast, no significant differences in hTLR2 activation were observed. Similarly, vaccine-induced activation of MonoMac-6 and monocyte-derived dendritic cells was strongest with the highest potency vaccine. Blocking of TLR2 and TLR4 showed that differences in antigen-presenting cell activation could be largely attributed to vaccine-dependent variation in hTLR4 signalling. Interestingly, this BvgASR-dependent decrease in hTLR4 activation coincided with a reduction in GlcN-modified lipopolysaccharides in these vaccines. Accordingly, expression of the lgmA-C genes, required for this glucosamine modification, was significantly reduced in bacteria exposed to sulfate. Together, these findings demonstrate that the BvgASR status of bacteria during wP vaccine preparation is critical for their hTLR4 activation capacity and suggest that including

  13. Improved detection of extended spectrum beta-lactamase (ESBL-producing Escherichia coli in input and output samples of German biogas plants by a selective pre-enrichment procedure.

    Directory of Open Access Journals (Sweden)

    Thorsten Schauss

    Full Text Available The presence of extended-spectrum beta-lactamase (ESBL-producing Escherichia coli was investigated in input (manure from livestock husbandry and output samples of six German biogas plants in 2012 (one sampling per biogas plant and two German biogas plants investigated in an annual cycle four times in 2013/2014. ESBL-producing Escherichia coli were cultured by direct plating on CHROMagar ESBL from input samples in the range of 100 to 104 colony forming units (CFU per g dry weight but not from output sample. This initially indicated a complete elimination of ESBL-producing E. coli by the biogas plant process. Detected non target bacteria were assigned to the genera Acinetobacter, Pseudomonas, Bordetella, Achromobacter, Castellaniella, and Ochrobactrum. A selective pre-enrichment procedure increased the detection efficiency of ESBL-producing E. coli in input samples and enabled the detection in five of eight analyzed output samples. In total 119 ESBL-producing E. coli were isolated from input and 46 from output samples. Most of the E. coli isolates carried CTX-M-type and/or TEM-type beta lactamases (94%, few SHV-type beta lactamase (6%. Sixty-four blaCTX-M genes were characterized more detailed and assigned mainly to CTX-M-groups 1 (85% and 9 (13%, and one to group 2. Phylogenetic grouping of 80 E. coli isolates showed that most were assigned to group A (71% and B1 (27%, only one to group D (2%. Genomic fingerprinting and multilocus sequence typing (MLST showed a high clonal diversity with 41 BOX-types and 19 ST-types. The two most common ST-types were ST410 and ST1210. Antimicrobial susceptibility testing of 46 selected ESBL-producing E. coli revealed that several isolates were additionally resistant to other veterinary relevant antibiotics and some grew on CHROMagar STEC but shiga-like toxine (SLT genes were not detected. Resistance to carbapenems was not detected. In summary the study showed for the first time the presence of ESBL-producing E

  14. Pertussis toxin analog with reduced enzymatic and biological activities is a protective immunogen.

    Science.gov (United States)

    Kimura, A; Mountzouros, K T; Schad, P A; Cieplak, W; Cowell, J L

    1990-01-01

    Bordetella pertussis TOX3201 has a 12-base-pair insertion in the S1 subunit gene of pertussis toxin (PTX), which encodes for a 4-amino-acid insertion between residues 107 and 108 of the mature S1 subunit (Black et al., Science 240:656-659, 1988). This mutant strain has been shown to secrete a holotoxin analog of PTX, designated CRM3201, with reduced ADP-ribosyltransferase activity. In the present study, we evaluated the biochemical, biological, and immunoprotective activities of purified CRM3201. Assay of enzymatic activities showed that CRM3201 had 20 to 30% of the ADP-ribosyltransferase activity and 55 to 60% of the NAD glycohydrolase activity of native PTX. CRM3201, however, had only 2 to 6% of the activity of PTX in clustering CHO cells, promoting leukocytosis, inducing histamine sensitization, and potentiating an anaphylactic response to bovine serum albumin. In contrast, activities associated with the B oligomer (binding to fetuin, hemagglutination of goose erythrocytes, and lymphocyte mitogen activity) were comparable to those of native PTX. Injection of BALB/c mice with CRM3201 mixed with Al(OH)3 elicited high titers of antibody to PTX (as measured by enzyme-linked immunosorbent assay), which neutralized a leukocytosis-promoting dose of PTX in these mice and neutralized PTX in a CHO cell assay. Passive transfer of the anti-CRM3201 antibody protected 20-day-old Swiss-Webster mice against a lethal aerosol challenge with B. pertussis 18323. Active immunization with CRM3201 significantly reduced lung colonization in adult BALB/c mice with a B. pertussis respiratory infection. These results demonstrate (i) that the reduced ADP-ribosyltransferase activity of CRM3201 is associated with reductions in certain biological and toxic activities of PTX (the enzymatic and biological activities are not, however, totally concordant); (ii) that CRM3201 possesses a functional B oligomer; and (iii) that CRM3201 can induce toxin-neutralizing antibodies which protect mice

  15. Infecções por vírus respiratórios em lactentes com suspeita clínica de coqueluche

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    Angela E. Ferronato

    2013-12-01

    Full Text Available OBJETIVO: avaliar a frequência das infecções por vírus respiratórios em lactentes hospitalizados com suspeita clínica de coqueluche e analisar suas características admissionais e evolutivas. MÉODOS: foi realizado um estudo de coorte histórica, em um serviço sentinela para coqueluche, no qual a pesquisa de vírus respiratórios também foi rotineira para os lactentes hospitalizados com problemas respiratórios. Foram incluídos todos os lactentes submetidos à notificação compulsória de suspeita de coqueluche. Foram realizadas pesquisas para Bordetela pertussis - BP (PCR/cultura e vírus respiratórios - VR (imunofluorescência. Foram excluídos os pacientes que haviam recebido macrolídeos previamente à internação. Os dados clínicos foram obtidos dos prontuários. RESULTADOS: dentre os 67 pacientes analisados, a pesquisa para BP foi positiva em 44% e para VR em 26%. Não houve identificação etiológica em 35% e em 5% houve codetecção de VR e BP. Todos os pacientes apresentaram características demográficas semelhantes. A presença de tosse seguida de guincho inspiratório ou cianose foi um forte preditor de coqueluche, assim como, leucocitose e linfocitose evidentes. Coriza e dispneia foram mais frequentes nas infecções virais. Houve suspensão do uso de macrolídeos em 40% dos pacientes com pesquisa positiva para VR e negativa para BP. CONCLUSÃO: os resultados sugerem que lactentes hospitalizados com suspeita de coqueluche podem apresentar infecção viral e a pesquisa etiológica pode possibilitar a redução do uso de macrolídeos em alguns casos. No entanto, salienta-se que o diagnóstico etiológico de infecção por vírus respiratórios, por si só, não exclui a possibilidade de infecção por Bordetella pertussis.

  16. Combination of pneumococcal surface protein A (PspA with whole cell pertussis vaccine increases protection against pneumococcal challenge in mice.

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    Maria Leonor S Oliveira

    Full Text Available Streptococcus pneumoniae is the leading cause of respiratory acute infections around the world. In Latin America, approximately 20,000 children under 5 years of age die of pneumococcal diseases annually. Pneumococcal surface protein A (PspA is among the best-characterized pneumococcal antigens that confer protection in animal models of pneumococcal infections and, as such, is a good alternative for the currently available conjugated vaccines. Efficient immune responses directed to PspA in animal models have already been described. Nevertheless, few low cost adjuvants for a subunit pneumococcal vaccine have been proposed to date. Here, we have tested the adjuvant properties of the whole cell Bordetella pertussis vaccine (wP that is currently part of the DTP (diphtheria-tetanus-pertussis vaccine administrated to children in several countries, as an adjuvant to PspA. Nasal immunization of BALB/c mice with a combination of PspA5 and wP or wP(low--a new generation vaccine that contains low levels of B. pertussis LPS--conferred protection against a respiratory lethal challenge with S. pneumoniae. Both PspA5-wP and PspA5-wP(low vaccines induced high levels of systemic and mucosal antibodies against PspA5, with similar profile, indicating no essential requirement for B. pertussis LPS in the adjuvant properties of wP. Accordingly, nasal immunization of C3H/HeJ mice with PspA5-wP conferred protection against the pneumococcal challenge, thus ruling out a role for TLR4 responses in the adjuvant activity and the protection mechanisms triggered by the vaccines. The high levels of anti-PspA5 antibodies correlated with increased cross-reactivity against PspAs from different clades and also reflected in cross-protection. In addition, passive immunization experiments indicated that antibodies played an important role in protection in this model. Finally, subcutaneous immunization with a combination of PspA5 with DTP(low protected mice against challenge with two

  17. Clinical presentation of pertussis in fully immunized children in Lithuania

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    Bernatoniene Genovaite

    2005-05-01

    Full Text Available Abstract Background In Lithuania, the vaccination coverage against pertussis is high. Nevertheless, there is a significant increase in pertussis cases in fully immunized children. The aim of our study was to determine the frequency of classical symptoms of laboratory confirmed pertussis and describe its epidemiology in children fully vaccinated against pertussis. Methods From May to December 2001, 70 children aged 1 month to 15 years, suffering from prolonged cough were investigated in the Centre of Paediatrics, Vilnius University Children's Hospital. The collected information included personal data, vaccination history, clinical symptoms of the current illness, and treatment before hospitalization. At the admission to the hospital blood samples were taken from all studied children for Bordetella pertussis IgM and IgA. Results A total of 53 (75.7% of the 70 recruited patients with prolonged cough showed laboratory evidence of pertussis. 32 of them were fully vaccinated with whole cell pertussis vaccine (DTP. The age of fully vaccinated patients varied from 4 to 15 years (average 10.9 ± 3.1; median 11. The time period between the last vaccination dose (fourth and the clinical manifestation of pertussis was 2.6–13 years (average 8.9 ± 3.0; median 9. More than half of the children before the beginning of pertussis were in contact with persons suffering from long lasting cough illness in the family, school or day-care center. The mean duration from onset of pertussis symptoms until hospitalization was 61.4 ± 68.3 days (range, 7 to 270 days; median 30. For 11 patients who had had two episodes (waves of coughing, the median duration of cough was 90 days, and for 21 with one episode 30 days (p Conclusion Fully vaccinated children fell ill with pertussis at the median of 11 years old, 9 years following pertussis vaccination. More than half of the children could catch pertussis at home, at school or day-care center. Clinical picture of pertussis in

  18. Effect of different detoxification procedures on the residual pertussis toxin activities in vaccines.

    Science.gov (United States)

    Yuen, Chun-Ting; Asokanathan, Catpagavalli; Cook, Sarah; Lin, Naomi; Xing, Dorothy

    2016-04-19

    Pertussis toxin (PTx) is a major virulence factor produced by Bordetella pertussis and its detoxified form is one of the major protective antigens in vaccines against whooping cough. Ideally, PTx in the vaccine should be completely detoxified while still preserving immunogenicity. However, this may not always be the case. Due to multilevel reaction mechanisms of chemical detoxification that act on different molecular sites and with different production processes, it is difficult to define a molecular characteristic of a pertussis toxoid. PTx has two functional distinctive domains: the ADP-ribosyltransferase enzymatic subunit S1 (A-protomer) and the host cell binding carbohydrate-binding subunits S2-5 (B-oligomer); and in this study, we investigated the effect of different detoxification processes on these two functional activities of the residual PTx in toxoids and vaccines currently marketed worldwide using a recently developed in vitro biochemical assay system. The patho-physiological activities in these samples were also estimated using the in vivo official histamine sensitisation tests. Different types of vaccines, detoxified by formaldehyde, glutaraldehyde or by both, have different residual functional and individual baseline activities. Of the vaccines tested, PT toxoid detoxified by formaldehyde had the lowest residual PTx ADP-ribosyltransferase activity. The carbohydrate binding results detected by anti-PTx polyclonal (pAb) and anti-PTx subunits monoclonal antibodies (mAb) showed specific binding profiles for toxoids and vaccines produced from different detoxification methods. In addition, we also demonstrated that using pAb or mAb S2/3 as detection antibodies would give a better differential difference between these vaccine lots than using mAbs S1 or S4. In summary, we showed for the first time that by measuring the activities of the two functional domains of PTx, we could characterise pertussis toxoids prepared from different chemical detoxification

  19. Incidence and reproduction numbers of pertussis: estimates from serological and social contact data in five European countries.

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    Mirjam Kretzschmar

    2010-06-01

    Full Text Available BACKGROUND: Despite large-scale vaccination programmes, pertussis has remained endemic in all European countries and has been on the rise in many countries in the last decade. One of the reasons that have been discussed for the failure of vaccination to eliminate the disease is continued circulation of the pathogen Bordetella pertussis by mostly asymptomatic and mild infections in adolescents and adults. To understand the impact of asymptomatic and undiagnosed infection on the transmission dynamics of pertussis we analysed serological data from five European countries in combination with information about social contact patterns from five of those countries to estimate incidence and reproduction numbers. METHODS AND FINDINGS: We compared two different methods for estimating incidence from individual data on IgG pertussis toxin (PT titres. One method combines the cross-sectional surveys of titres with longitudinal information about the distribution of amplitude and decay rate of titres in a back-calculation approach. The second method uses age-dependent contact matrices and cross-sectional surveys of IgG PT titres to estimate a next generation matrix for pertussis transmission among age groups. The next generation approach allows for computation of basic reproduction numbers for five European countries. Our main findings are that the seroincidence of infections as estimated with the first method in all countries lies between 1% and 6% per annum with a peak in the adolescent age groups and a second lower peak in young adults. The incidence of infections as estimated by the second method lies slightly lower with ranges between 1% and 4% per annum. There is a remarkably good agreement of the results obtained with the two methods. The basic reproduction numbers are similar across countries at around 5.5. CONCLUSIONS: Vaccination with currently used vaccines cannot prevent continued circulation and reinfection with pertussis, but has shifted the bulk

  20. Regulation of sigma S degradation in Salmonella enterica var typhimurium: in vivo interactions between sigma S, the response regulator MviA(RssB) and ClpX.

    Science.gov (United States)

    Moreno, M; Audia, J P; Bearson, S M; Webb, C; Foster, J W

    2000-04-01

    The alternate sigma factor sigmaS plays an important role in the survival of Salmonella typhimurium following sudden encounters with a variety of stress conditions. The level of sigmaS is very low in rapidly growing cells but dramatically increases as those cells encounter environmental stress or enter into stationary phase. This increase is due in large measure to the stabilization of sigmaS protein against degradation by the ClpXP protease. The MviA protein, also known as RssB or SprE in Escherichia coli, is a putative member of a two component signal transduction system that plays a central role in facilitating sigmaS degradation by ClpXP. In contrast to most two-component systems, MviA does not appear to regulate gene expression but is believed to interact directly with sigmaS and somehow facilitate degradation. We now provide evidence that MviA(RssB) directly interacts both with sigmaS and ClpX in vivo, presumably enabling presentation of sigmaS to the ClpP protease. Interactions were demonstrated using a bacterial two-hybrid system in which sigmaS, MviA, and ClpX were fused to separate moieties of Bordetella pertussis CyaA (adenylate cyclase). Paired hybrid plasmids containing Cya'-MviA/RpoS-'Cya or Cya'-MviA/ClpX-'Cya successfully reconstituted adenylate cyclase activity in both S. typhimurium and E. coli. However, no direct interactions were detected between ClpX and RpoS. A second series of experiments has indicated that the interaction between MviA and sigmaS requires the N-terminus but not the C-terminus of MviA. Cellular levels of MviA appear to be very low in the cell based on lacZ fusion, Western blot and Northern blot analyses suggesting a catalytic role for MviA in sigmaS degradation. Mutagenesis of MviA residue D58, a canonical residue subject to phosphorylation in many two-component systems, decreased the ability of MviA to facilitate sigmaS turnover in vivo confirming that phosphorylation of MviA increases MviA activity. PMID:10939250

  1. [Bacteriological and virological status in upper respiratory tract infections of cats (cat common cold complex)].

    Science.gov (United States)

    Adler, Kerstin; Radeloff, Isabel; Stephan, Bernd; Greife, Heinrich; Hellmann, Klaus

    2007-01-01

    Between October 2002 and January 2005,460 bacteriological samples from cats with an acute upper respiratory tract infection were analysed in clinical field studies in two accredited laboratories in Germany. Oropharyngeal swabs were taken from these cats and sent to the laboratories for routine diagnostics. In the swab samples of 460 cats 382 bacteria strains were isolated. The following bacteria were isolated most frequently: Pasteurella spp. (32.5 %), Staphylococcus spp. (18.5 %), Escherichia coli (17.0 %), Streptococcus spp. (9.1 %), Pseudomonas spp. (6.9 %) and Klebsiella spp. (3.0 %). Bordetella bronchiseptica was found in 0.4 % of the animals To evaluate possible regional and time influences, the animals were split into three populations: 1: Germany, laboratory A; 2: Germany, laboratory B; 3: France and Belgium, laboratory B. In population 1 an 2 Pasteurella spp. were found most frequently with 42.2 % and 36.5 %, respectively. The second most frequently isolated bacterial species were Staphylococcus spp. with 14.1 % and 21.4 % and E. coli with 13.6 % and 17.5 % respectively. In population 3 Staphylococcus spp., E. coli (20 % each) and Pasteurella spp. (18.5 %) were isolated at almost the same frequency. Virological parameter were additionally analysed in 328 cats (population 2 and 3). Serum samples were analysed for antibodies specific for Feline Calicivirus (FCV) and Feline Immunodeficiency Virus (FIV) and for Feline Leukaemia Virus (FeLV) antigen. Oropharyngeal swabs were analysed for Feline Herpesvirus (FHV) by using PCR. Calicivirus-specific antibodies were found in 99.6 % of the cats of population 2 and in 100 % of the animals in population 3. Herpesvirus was detected in 15.3 % and 23.3 % of the cats, respectively. FeLV-Antigen was found in 0.4 % of the animals in population 2 and in 10.1 % of the cats in population 3, while FIV-antibodies were identified in 8.7 % of the animals of population 2 and in 6.1 % of the cats of population 3. In total FHV was

  2. Effects of realgar on stress proteins, inflammatory mediators, and complement in brain tissue and serum of rats with inflammatory brain injury

    Institute of Scientific and Technical Information of China (English)

    Yishan Tang; Ningsheng Wang; Yinqing Zhang; Shaomei Ye; Weiping Ou

    2008-01-01

    BACKGROUND: The Chinese herbal compound realgar exerts detoxification effects as an adjuvant. It is suggested that realgar exerts detoxification via the following pathways: in the pathological state, realgar corrects the oxidative stress state by increasing stress levels, activating some endogenous protective factors and antagonizing the excessive release of inflammatory factors, as well as inhibiting complement activation.OBJECTIVE: To observe the changes in stress proteins, inflammatory mediators, and complement in the brain tissue and serum of rats with inflammatory brain injury, which have been treated with thc Chinese herbal compound Angong Niuhuang, and to compare the efficacy of Angong Niuhuang with that of realgar,to verify the mechanism of action of realgar.DESIGN, TIME AND SETTING: Randomized, controlled, cytological experiment, performed in the Institute of Clinical Pharmacology, Guangzhou University of Traditional Chinese Medicine in March 2006.MATERIALS: Thirty-six healthy, male, Sprague Dawley rats received 250 U/kg Bordetella pertussis via the common carotid artery within 15 seconds to induce inflammatory brain injury. Reagents and kits were as follows: Realgar and Angong Niuhuang powder (Foshan Second Pharmaceutical Factory, China), Bordetella pertussis diagnostic antigen (National Institute for the Control of Pharmaceutical and Biological Products,China), heat shock protein 70 (HSP70) enzyme-labeled immunosorbent assay (ELISA) kit (Stressgen, USA),tumor necrosis factor-α (TNF-α) ELISA kit (Biosource, USA), nitric oxide synthase (NOS) kit,Coomassie brilliant blue protein kit (Nanjing Jiancheng Bioengineering Co.,Ltd., China), and complements C3 and C4 (Shanghai Kehua Dongling Diagnositic Products Co.,Ltd., China),METHODS: Thirty-six rats were randomly and evenly divided into the following six groups: normal control,model, high-, middle-, and low-dose realgar-treated, and Angong Niuhuang-treated groups. At one hour prior to establishing the model

  3. Primera incursión en la obtención de curieles libre de patógenos específicos en Cuba

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    Bárbara O González Navarro

    2014-06-01

    Full Text Available Título corto: Primera incursión en la obtención de curieles SPF en CubaTítulo en ingles: First foray into the production of specific pathogen-free guinea pigs in CubaResumen: Resultados confiables y económicos solo son obtenidos cuando los animales de experimentación son aislados de factores ambientales y biológicos, implantándose en el biomodelo una microbiota normal, lejos de la presencia de microorganismos patógenos. El objetivo de la investigación fue obtener curieles libre de patógenos específicos por cesárea aséptica, mantenidos en aisladores y alimentados con dietas estériles. Se realizaron 26 histerectomías. Los animales fueron alimentados con una fórmula modificada (L-477 en forma de papilla hasta los 21 días y permanentemente después la C-484 sólida y granulada, esterilizadas a 121oC/20 minutos o a 1,5Mrad. Además fueron suplementados con vitamina C y B1. El forraje o heno fue consumido a partir de la primera generación. La microbiota gastrointestinal se administró por vía oral en 0,5 ml de una dilución de 10-6/g de contenido de la porción final del íleon, ciego y principio del colon de curieles, a las 24 y 48 horas del nacimiento. Se utilizó para el monitoreo microbiológico caldo Tioglicolato, caldo Triptona Soya y caldo Saboraud incubados aeróbicamente a temperatura de 55, 37y 25oC respectivamente. Se obtuvieron 51 neonatos. La mortalidad más alta se registró entre los primeros 10 días de edad (58,8%. Se lograron 12 animales (3 machos y 9 hembras, 6 de las hembras se reprodujeron aproximadamente a los 9 meses de edad, lográndose 11 crías por parto normal. La metodología aplicada permitió obtener curieles libres de Salmonella sp, Pasteurella sp, Streptococcus del tipo A y C, Bordetella bronchiseptica, Toxoplasma gondii, Virus Sendai y parásitos internos y externos.Palabras clave: curieles, libre de patógenos específicos, aisladores, histerectomía y dietas estériles.Abstract: Reliable and

  4. Vacina acelular contra pertússis para adolescentes Acellular pertussis vaccine for adolescents

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    Aroldo P. de Carvalho

    2006-07-01

    Full Text Available OBJETIVOS: A utilização da vacina de células inteiras contra coqueluche levou a uma redução significativa na incidência da enfermidade na criança. Essa mudança no perfil epidemiológico resultou em aumento no número de casos em adolescentes e adultos, conseqüente à perda da imunidade conferida pela doença ou por vacina após cerca de 10 anos, e em lactentes não imunizados ou incompletamente imunizados. O licenciamento da vacina tríplice bacteriana contra difteria, tétano e coqueluche acelular, com formulação específica para maiores de 10 anos de idade (dTpa, apontou para a possibilidade do controle da coqueluche na população das faixas etárias mais acometidas nos últimos anos. FONTE DE DADOS: As informações foram coletadas na base de dados MEDLINE. A pesquisa foi limitada ao período compreendido entre janeiro de 1995 a janeiro de 2006. SÍNTESE DOS DADOS: Estão licenciadas em alguns países duas vacinas dTpa para a faixa etária maior de 10 anos de idade, uma delas contendo cinco componentes imunogênicos da Bordetella pertussis: toxina pertússis, hemaglutinina filamentosa, fimbrias 2 e 3 e pertactina, e a outra contendo três componentes: pertactina, hemaglutinina filamentosa e toxina pertússis inativada, sendo esta a única apresentação licenciada até o momento no Brasil. Embora a composição das duas vacinas seja diferente, os estudos mostram que a imunogenicidade e a eficácia são semelhantes. Entretanto, alguns autores enfatizam que existem dificuldades para a realização de uma avaliação mais precisa da resposta imunológica à vacina e sua duração. Vários países já recomendam de rotina o uso da vacina dTpa para adolescentes. O Canadá ampliou a população alvo até 54 anos de idade. A orientação é de que esse grupo receba uma dose da vacina como reforço do esquema básico de imunização. Isso é fundamentado em resultados de estudos que mostram que a duração da imunidade induzida pela

  5. Triazenos e atividade antibacteriana Triazenes and antibacterial activity

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    Manfredo Hörner

    2008-09-01

    Full Text Available 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 sp., Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Bacillus cereus, Corynebacterium sp., Rhodococcus sp., Salmonella sp., Serratia marcescens, Morganella morganii, Enterobacter cloacae, Shigella flexneri, Shigella sonnei, Shigella sp., Klebsiella pneumoniae, ESBL Klebsiella oxytoca 14, ESBL Klebsiella pneumoniae 23, ESBL Klebsiella pneumoniae 24, ESBL Klebsiella pneumoniae 25, ESBL Escherichia coli 26, ESBL Klebsiella pneumoniae 27, ESBL Klebsiella pneumoniae 31, ESBL Escherichia coli 32, ESBL Klebsiella pneumoniae 37 e ESBL Escherichia coli 38. A maior atividade foi evidenciada para o composto 1-metil-3-(p-carboxifeniltriazeno 1-óxido (2 contra Streptococcus agalactiae (CIM =16 µg/mL e CBM = 32 µg/mL. Os compostos 1-fenil-3-(4-nitrofeniltriazeno-1-óxido (9, 1-(4-nitrofenil-3-(4-carboxifeniltriazeno (10 e 1-(4-acetilaminofenil-3-(4-carboxifeniltriazeno (11 apresentaram CIMs de 32 a 64 µg/mL frente a S. edipermidis, S. saprophyticus, Corynebacterium sp. e E. cloacae. Os compostos 1-metil-3-feniltriazeno-1-óxido (1 , bis-1,3-(4-acetiloximatriazeno (3, bis-1,3 (4-acetilfeniltriazeno (4, 1-(2-fluorfenil-3-(4-acetilfeniltriazeno (5, 1,3-(3-hidroxi-difeniltriazenido(piridil(bis-oxo-vanádio (12, 1-(3-nitrofenil-3-feniltriazeno (14, 1-(4-nitrofenil-3-benziltriazeno (15 apresentaram CIM = 128 µg/mL frente a S. aureus ATCC 25923, P. aeruginosa ATCC 27853, A. lwoffii, Micrococcus sp., S. epidermidis, S

  6. 兔多杀性巴氏杆菌外膜蛋白A(OmpA)重组蛋白单克隆抗体的制备及潜在应用%Preparation and Potential Application of Monoclonal Antibody against Pasteurella multocida Outer Membrane Protein A (OmpA) Recombinant Protein

    Institute of Scientific and Technical Information of China (English)

    刘燕; 庞安娜; 韦强; 肖琛闻; 鲍国连; 季权安; 钱微

    2012-01-01

    The aim of this study was to prepare monoclonal antibody (McAb)against Pasteurella multocida. The DNA fragment encoding the mature domain of P. multocida outer membrane protein A (OmpA) was amplified from the genomic DNA and sub-cloned into pET28a (+) expression vector, 37.6 kD rOmpA fusion protein was expressed mainly as an insoluble protein, optimal sohibilization of the recombinant protein was obtained using 8 mol/L urea in lysis buffer. BALB/c mice (Mus musculus) were subcutaneously injected with 100 μg of P. multocida OmpA emulsified by equivolumminal freund's complete adjuvant at the age of 6-8 weeks. Thereafter they were boosted two times with 200 μg of P. multocida OmpA emulsified by Freund's incomplete adjuvant at intervals of three weeks. The spleen cells of BALB/ c mice immunized with recombinant Pm OmpA were collected and infused with SP2/ 0 cell. Sebsequently four hybridoma cell strains were obtained by indirect enzyme linked immunosorbent assay (ELISA). The ELISA titers of antibodies in culture supernatant were 1:128,1:128,1:256 and 1:128, respectively, and ascites titers were 1:6 400,1:6 400,1: 12 800 and 1:6 400, respectively. The McAbs did not cross-react with other gram-negative and gram-positive bacterial pathogens, including E. coli, Bordetella bmnchiseptka, Pseudomonas aeruginosa and slaphylococcus. High titer McAbs were secreted from the hybridoma cells after repeat freezing. The result of Western blotting assay showed that the four Mabs could react with Pm OmpA protein specifically. ELISA test revealed that the 2A2 McAb belonged to the subtype of IgG2b, with a concentration was 130 u-g/mL after protein A affinity purification. The purified 2A2 McAb was selected by Western blot and IFA assays. The result indicated that the McAb could react with the Pm isolate strain. The success of this study has built up a solid base for developing a novel diagnostic methodology to the Pasteurella multocida infection in rabbits.%制

  7. Expression of myelin basic protein in rat optic nerve with experimental allergic encephalomyelitis%实验性变态反应性脑脊髓炎大鼠视神经内髓鞘碱性蛋白表达的变化

    Institute of Scientific and Technical Information of China (English)

    曹小鹏; 高晓唯; 曹芃; 雷英; 刘李平

    2012-01-01

    的延长,MBP/β-actin值在免疫组大鼠视神经组织中的表达逐渐减少,免疫后12d,M BP/β-actin值小于正常对照组,差异有统计学意义( t=4.639,P<0.05),免疫后18 d MBP/β-actin值最低,与正常对照组比较差异有统计学意义(t=8.427,P<0.01).结论 EAE大鼠视神经组织存在MBP的降解,提示视神经炎是一种视神经的原发性脱髓鞘病变.%Background Optic neuritis is closely associated with multiple sclerosis (MS).Its pathogenesis is uncompletely clear,and less basic researches are carried out at home and abroad. Objective This study was to reveal the expression of myelin basic protein (MBP) in the optic nerve of rat with experimental allergic encephalomyelitis (EAE) and to provide a theoretical evidence for the research of the relationship of optic neuritis with MS. Methods Fifty clean Wistar rats were randomized into the control group and immune 8,12,18 and 25 days groups.Myelencephalon was collected from 5 guinea pigs to prepare the homogenate and mixed with the isovolumetric complete Freud' s adjuvant (CFA).The 0.5 ml mixed antigen emulsifier was subcutancously injected into the 4 maps together with Bordetella pertussis 0.2 ml under the cutancous of dorsalis pedis at 0 and 48 hours to induce the EAE.Behavior of the rats was evaluated to score the neurological function.The optical nerve sections were prepared 8,12,18 and 25 days after immunology for the histopathological examination,and immunochemistry and Western blot were used to detect the expression of MBP in optic nerve.The use of the animals complied with the Regulation for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission. Results The disorder of motor nerve was seen 12 days following the immune,and the clinical neural functional scores were significantly higher 12 day and peaked on 18 days myelination and then gradually reduced.The histopathological examination showed that the irregular alignment