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Sample records for adopt haemophilus influenzae

  1. About Haemophilus influenzae Disease

    Science.gov (United States)

    ... one that most people are familiar with is Haemophilus influenzae type b, or Hib. There’s a vaccine that can prevent disease caused by Hib, but not the other types of Haemophilus influenzae . Types of Infection Haemophilus influenzae Can Cause Infections ...

  2. Haemophilus Influenzae Type b

    Science.gov (United States)

    ... Issues Listen Español Text Size Email Print Share Haemophilus Influenzae type b Page Content Article Body If you’re like many parents, you may have been unfamiliar with Haemophilus influenzae type b (Hib) infections until your pediatrician recommended a vaccine ...

  3. Accelerating policy decisions to adopt haemophilus influenzae type B vaccine: a global, multivariable analysis.

    Directory of Open Access Journals (Sweden)

    Jessica C Shearer

    2010-03-01

    Full Text Available BACKGROUND: Adoption of new and underutilized vaccines by national immunization programs is an essential step towards reducing child mortality. Policy decisions to adopt new vaccines in high mortality countries often lag behind decisions in high-income countries. Using the case of Haemophilus influenzae type b (Hib vaccine, this paper endeavors to explain these delays through the analysis of country-level economic, epidemiological, programmatic and policy-related factors, as well as the role of the Global Alliance for Vaccines and Immunisation (GAVI Alliance. METHODS AND FINDINGS: Data for 147 countries from 1990 to 2007 were analyzed in accelerated failure time models to identify factors that are associated with the time to decision to adopt Hib vaccine. In multivariable models that control for Gross National Income, region, and burden of Hib disease, the receipt of GAVI support speeded the time to decision by a factor of 0.37 (95% CI 0.18-0.76, or 63%. The presence of two or more neighboring country adopters accelerated decisions to adopt by a factor of 0.50 (95% CI 0.33-0.75. For each 1% increase in vaccine price, decisions to adopt are delayed by a factor of 1.02 (95% CI 1.00-1.04. Global recommendations and local studies were not associated with time to decision. CONCLUSIONS: This study substantiates previous findings related to vaccine price and presents new evidence to suggest that GAVI eligibility is associated with accelerated decisions to adopt Hib vaccine. The influence of neighboring country decisions was also highly significant, suggesting that approaches to support the adoption of new vaccines should consider supply- and demand-side factors.

  4. Haemophilus influenzae biotype VIII.

    OpenAIRE

    Sottnek, F O; Albritton, W L

    1984-01-01

    Six Haemophilus influenzae strains could not be classified as biotypes I through VII. The strains were indole, urease, and ornithine decarboxylase negative. We propose that they be classified as biotype VIII, a previously unreported biotype.

  5. Haemophilus influenzae biotype VII.

    OpenAIRE

    Gratten, M

    1983-01-01

    A hitherto unreported biotype of Haemophilus influenzae is described. The isolate is noncapsulate and fails to decarboxylate ornithine or hydrolyze urea but is a strong indole producer. Its frequency is low. It is suggested that this newly recognized biotype of H. influenzae be designated biotype VII.

  6. Invasive Disease Caused by Nontypeable Haemophilus influenzae

    Science.gov (United States)

    de Jonge, Marien I.

    2015-01-01

    The incidence of severe Haemophilus influenza infections, such as sepsis and meningitis, has declined substantially since the introduction of the H. influenzae serotype b vaccine. However, the H. influenzae type b vaccine fails to protect against nontypeable H. influenzae strains, which have become increasingly frequent causes of invasive disease, especially among children and the elderly. We summarize recent literature supporting the emergence of invasive nontypeable H. influenzae and describe mechanisms that may explain its increasing prevalence over the past 2 decades. PMID:26407156

  7. NNDSS - Table II. Giardiasis to Haemophilus influenza

    Data.gov (United States)

    U.S. Department of Health & Human Services — NNDSS - Table II. Giardiasis to Haemophilus influenza - 2015.In this Table, provisional cases of selected notifiable diseases (≥1,000 cases reported during the...

  8. NNDSS - Table II. Giardiasis to Haemophilus influenza

    Data.gov (United States)

    U.S. Department of Health & Human Services — NNDSS - Table II. Giardiasis to Haemophilus influenza - 2014. In this Table, all conditions with a 5-year average annual national total of more than or equals 1,000...

  9. NNDSS - Table II. Giardiasis to Haemophilus influenza

    Data.gov (United States)

    U.S. Department of Health & Human Services — NNDSS - Table II. Giardiasis to Haemophilus influenza - 2016. In this Table, provisional* cases of selected†notifiable diseases (≥1,000 cases reported during...

  10. Protein sources of heme for Haemophilus influenzae.

    OpenAIRE

    Stull, T L

    1987-01-01

    Although Haemophilus influenzae requires heme for growth, the source of heme during invasive infections is not known. We compared heme, lactoperoxidase, catalase, cytochrome c, myoglobin, and hemoglobin as sources of heme for growth in defined media. The minimum concentration of heme permitting unrestricted growth of strain E1a, an H. influenzae type b isolate from cerebrospinal fluid, was 0.02 micrograms/ml. Using molar equivalents of heme as lactoperoxidase, catalase, cytochrome c, myoglobi...

  11. Population Structure in Nontypeable Haemophilus influenzae

    OpenAIRE

    LaCross, Nathan C.; Marrs, Carl F.; Gilsdorf, Janet R.

    2012-01-01

    Nontypeable Haemophilus influenzae (NTHi) frequently colonize the human pharynx asymptomatically, and are an important cause of otitis media in children. Past studies have identified typeable H. influenzae as being clonal, but the population structure of NTHi has not been extensively characterized. The research presented here investigated the diversity and population structure in a well-characterized collection of NTHi isolated from the middle ears of children with otitis media or the pharyng...

  12. Plasmid recombination in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.

    1982-01-01

    DNA recombination in exponential phase and competent Haemophilus influenzae was measured by an electron microscopic assay that relies on the conversion of plasmid RSF0885 monomers into multimeric forms. Dimer circles were present at a frequency of 2% in plasmid preparations from competent Rd (wild-type) cells; multimers were present at a frequency of 0.2% in preparations from exponential phase cells. Thus, plasmid recombination was stimulated in competent cells. Multimer formation occurred efficiently in cells of the transformation defective mutant rec2, implying that the rec2 gene product is not required for plasmid recombination. However, the absence of multimer plasmids in preparations from competent cells of the transformation defective mutant rec1 suggests that the rec1 gene product is required. Digestion of purified plasmids with restriction endonuclease PvuII, which makes a single cut in the monomer, revealed the presence of recombination intermediates composed of two linear plasmids joined to form two pairs of arms resembling the Greek letter chi. Length measurements of these arms taken from a population of recombination intermediates gave evidence that the plasmids were joined at sites of homology. The distributions of individual DNA strands, at the intersections of the four arms, could be resolved in some recombination intermediates and were of two types. The first type of junction appeared as a single-stranded arm appended to each corner. The second type of junction consisted of a single strand of DNA linking the two linear plasmids at a site of homology. The single-stranded linker was frequently situated at the edge of a short gap on one of the plasmids in the pair. The fine structures of the recombinational joints have been interpreted in terms of previously proposed models of recombination.

  13. Invasive Disease Caused by Nontypeable Haemophilus Influenzae

    Centers for Disease Control (CDC) Podcasts

    2015-11-12

    Dr. Elizabeth Briere discusses Nontypeable Haemophilus influenzae which causes a variety of infections in children and adults.  Created: 11/12/2015 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID).   Date Released: 11/17/2015.

  14. Transformation of Haemophilus influenzae by recombinant molecules

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, J.K.; McCarthy, D.R.; Clayton, N.L.

    1981-01-01

    A gene library of Haemophilus influenzae DNA has been constructed by ligating chromosomal DNA from cells resistant to a number of antibiotics, together with DNA of the H. influenzae plasmid RSF0885. Before ligation both DNAs were cut with the enzyme PvuII. The ligated DNA was allowed to enter competent H. influenzae sensitive to the antibiotics and selection was made for resistance to ampicillin, conferred by the plasmid RSF0885 DNA. Plasmids conferring resistance to various other antibiotics, as well as to ampicillin, have been obtained by this procedure and subsequent selection for chromosomal markers.

  15. Haemophilus influenzae Type b (Hib) vaccine - what you need to know

    Science.gov (United States)

    ... taken in its entirety from the CDC Hib (Haemophilus Influenzae Type b) Vaccine Information Statement (VIS): www.cdc. ... statements/hib.pdf . CDC review information for Hib (Haemophilus Influenzae Type b) VIS: Page last reviewed: April 2, ...

  16. Long-term mortality in children diagnosed with Haemophilus influenzae meningitis: a Danish nationwide cohort study

    DEFF Research Database (Denmark)

    Roed, Casper; Engsig, Frederik Neess; Omland, Lars Haukali;

    2011-01-01

    The long-term mortality in children diagnosed with Haemophilus influenzae meningitis is poorly documented.......The long-term mortality in children diagnosed with Haemophilus influenzae meningitis is poorly documented....

  17. Identifying Haemophilus haemolyticus and Haemophilus influenzae by SYBR Green real-time PCR.

    Science.gov (United States)

    Latham, Roger; Zhang, Bowen; Tristram, Stephen

    2015-05-01

    SYBR Green real time PCR assays for protein D (hpd), fuculose kinase (fucK) and [Cu, Zn]-superoxide dismutase (sodC) were designed for use in an algorithm for the identification of Haemophilus influenzae and H. haemolyticus. When tested on 127 H. influenzae and 60 H. haemolyticus all isolates were identified correctly. PMID:25753676

  18. Evaluation of new biomarker genes for differentiating Haemophilus influenzae from Haemophilus haemolyticus.

    Science.gov (United States)

    Theodore, M Jordan; Anderson, Raydel D; Wang, Xin; Katz, Lee S; Vuong, Jeni T; Bell, Melissa E; Juni, Billie A; Lowther, Sara A; Lynfield, Ruth; MacNeil, Jessica R; Mayer, Leonard W

    2012-04-01

    PCR detecting the protein D (hpd) and fuculose kinase (fucK) genes showed high sensitivity and specificity for identifying Haemophilus influenzae and differentiating it from H. haemolyticus. Phylogenetic analysis using the 16S rRNA gene demonstrated two distinct groups for H. influenzae and H. haemolyticus. PMID:22301020

  19. Evaluation of New Biomarker Genes for Differentiating Haemophilus influenzae from Haemophilus haemolyticus

    OpenAIRE

    Theodore, M. Jordan; Anderson, Raydel D.; Wang, Xin; Katz, Lee S.; Vuong, Jeni T.; Bell, Melissa E.; Juni, Billie A.; Sara A Lowther; Lynfield, Ruth; MacNeil, Jessica R.; Leonard W. Mayer

    2012-01-01

    PCR detecting the protein D (hpd) and fuculose kinase (fucK) genes showed high sensitivity and specificity for identifying Haemophilus influenzae and differentiating it from H. haemolyticus. Phylogenetic analysis using the 16S rRNA gene demonstrated two distinct groups for H. influenzae and H. haemolyticus.

  20. Plasmid containing a DNA ligase gene from Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.; Griffin, K.; Setlow, J.K.

    1984-05-01

    A ligase gene from Haemophilus influenzae was cloned into the shuttle vector pDM2. Although the plasmid did not affect X-ray sensitivity, it caused an increase in UV sensitivity of the wild-type but not excision-defective H. influenzae and a decrease in UV sensitivity of the rec-1 mutant. 14 references, 2 figures.

  1. Plasmid cloning vehicle for Haemophilus influenzae and Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.; Clayton, N.L.; Setlow, J.K.

    1982-09-01

    A new plasmid cloning vehicle (pDM2) was used to introduce a library of Haemophilus influenzae chromosomal fragments into H. influenzae. Transformants of the higly recombination-defective rec-1 mutant were more likely to contain exclusively recombinant plasmids after exposure to ligated DNA mixtures than was the wild type. pDM2 could replicate in Escherichia coli K-12.

  2. [Two cases of invasive Haemophilus influenzae type f infection

    DEFF Research Database (Denmark)

    Nielsen, J.D.; Lind, J.W.; Bruun, B.

    2009-01-01

    Two cases of invasive Haemophilus influenzae type f infection are presented: a three-week-old boy with meningitis and a 62-year-old woman with arthritis and bacteremia. Since 1993 vaccination against H. influenzae type b (Hib) has been offered to Danish children. The result has been a remarkable...

  3. To tilfaelde af invasive infektioner med Haemophilus influenzae type f

    DEFF Research Database (Denmark)

    Lind, Jens Wentzel; Bruun, Britta; Nielsen, Jette Dettmann;

    2009-01-01

    Two cases of invasive Haemophilus influenzae type f infection are presented: a three-week-old boy with meningitis and a 62-year-old woman with arthritis and bacteremia. Since 1993 vaccination against H. influenzae type b (Hib) has been offered to Danish children. The result has been a remarkable...

  4. Haemophilus haemolyticus: A Human Respiratory Tract Commensal to Be Distinguished from Haemophilus influenzae

    DEFF Research Database (Denmark)

    Murphy, T.F.; Brauer, A.L.; Sethi, S.;

    2007-01-01

    Background. Haemophilus influenzae is a common pathogen in adults with chronic obstructive pulmonary disease (COPD). In a prospective study, selected isolates of apparent H. influenzae had an altered phenotype. We tested the hypothesis that these variant strains were genetically different from.......Results. Variant strains were determined to be Haemophilus haemolyticus by means of 4 independent methods. Analysis of 490 apparent H. influenzae strains, identified by standard methods, revealed that 39.5% of sputum isolates and 27.3% of nasopharyngeal isolates were H. haemolyticus. Isolates obtained from...... typical H. influenzae.Methods. A prospective study of adults with COPD was conducted. Strains of apparent H. influenzae obtained from a range of clinical sources were evaluated by ribosomal DNA sequence analysis, multilocus sequence analysis, DNA-DNA hybridization, and sequencing of the conserved P6 gene...

  5. Transcriptional Profile of Haemophilus influenzae: Effects of Iron and Heme

    OpenAIRE

    Whitby, Paul W.; VanWagoner, Timothy M.; Seale, Thomas W.; Morton, Daniel J; Stull, Terrence L.

    2006-01-01

    Haemophilus influenzae requires either heme or a porphyrin and iron source for growth. Microarray studies of H. influenzae strain Rd KW20 identified 162 iron/heme-regulated genes, representing ∼10% of the genome, with ≥1.5-fold changes in transcription in response to iron/heme availability in vitro. Eighty genes were preferentially expressed under iron/heme restriction; 82 genes were preferentially expressed under iron/heme-replete conditions.

  6. [Severe Haemophilus influenzae b infection in healthy male adult

    DEFF Research Database (Denmark)

    Vilmar, A.C.; Gjorup, I.; David, Kim Peter

    2008-01-01

    Haemophilus influenzae b (Hib) can be the cause of serious infections, and is mainly observed affecting children and immuno-compromised patients. We report a case of a healthy 49-year old male with a severe Hib infection complicated by septicaemia, meningitis and anuria. The risk of invasive Hib...

  7. Heat sensitivity of Haemophilus influenzae containing defective prophage

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, J.K.

    1977-12-01

    Strains of Haemophilus influenzae that carry a defective prophage are more sensitive to heat than is a strain that does not, even in the presence of a rec-I mutation, which normally renders prophage noninducible. The prophage of HPlcl, a nondefective phage, does not affect the heat sensitivity.

  8. Haemophilus influenzae triggers autophagy in HEp-2 cells.

    Science.gov (United States)

    Espinoza-Mellado, María del Rosario; Reyes-Picaso, Carolina; Garcés-Pérez, Miriam S; Jardón-Serrano, Cynthia V; López-Villegas, Edgar O; Giono-Cerezo, Silvia

    2016-03-01

    The MAP-LC3 system regulates the intracellular formation of autophagy-associated vacuoles. These vacuoles contain the LC3 protein; thus it has been utilized as a marker to identify autophagosomes. The aim of our study was to investigate whether Haemophilus influenzae strains and their supernatants could activate autophagy in human larynx carcinoma cell line (HEp-2). We demonstrate that higher expression of the LC3B-II protein was induced, particularly by nontypeable Haemophilus influenzae (NTHi) 49766 and by supernatants, containing <50 kDa proteins, of both strains. Ultrastructural studies demonstrate vacuoles with a double membrane and/or membrane material inside, showing similar features to those of autophagic vacuoles. Together, our findings demonstrate that H. influenzae strains and their supernatants trigger an autophagic process.

  9. Haemophilus influenzae type f meningitis in a previously healthy boy

    DEFF Research Database (Denmark)

    Ronit, Andreas; Berg, Ronan M G; Bruunsgaard, Helle;

    2013-01-01

    Non-serotype b strains of Haemophilus influenzae are extremely rare causes of acute bacterial meningitis in immunocompetent individuals. We report a case of acute bacterial meningitis in a 14-year-old boy, who was previously healthy and had been immunised against H influenzae serotype b (Hib). Th......). The causative pathogen was identified as H influenzae serotype f (Hif), and was successfully treated with ceftriaxone. An immunological evaluation revealed transient low levels of immunoglobulins but no apparent immunodeficiency was found 2 years after the clinical insult....

  10. Molecular basis of antimicrobial resistance in non-typable Haemophilus influenzae.

    Science.gov (United States)

    Sánchez, L; Leranoz, S; Puig, M; Lorén, J G; Nikaido, H; Viñas, M

    1997-09-01

    Strains of the facultative anaerobe Haemophilus influenzae, both type b and non typable strains, are frequently multiresistant. The measurement of the antibiotic permeability of Haemophilus influenzae outer membrane (OM) shows that antibiotics can cross through the OM easily. Thus, enzymatic activity or efflux pumps could be responsible for multiresistance. An efflux system closely related to AcrAB of Escherichia coli is present in Haemophilus influenzae. However, their role in multiresistance seems irrelevant. Classical mechanisms such as plasmid exchange seems to be playing a major role in the multidrug resistance in Haemophilus influenzae.

  11. Non-Type B Haemophilus Influenzae Meningitis: A Case Report

    Directory of Open Access Journals (Sweden)

    Fatma Deniz Aygun

    2016-02-01

    Full Text Available Haemophilus influenza is one of the most common cause of bacterial meningitis in children. H.influenzae, especially type b (Hib serotype causes invasive infections in children under five years of age. The widespread use of Hib conjugate vaccines has led to a dramatic decline in the incidence of invasive Hib infections. But, the invasive diseases are still reported, particularly nontypeable H. influenzae (noncapsulated remain as an important pathogen. However, there is no evidence that nontypeable H. influenzae infections have increased in frequency. Nontypeable H. Ižnfluenzae serotype is encountered as a cause of acute bacterial meningitis among all ages. In this paper, we present to draw attention to the causative bacterium, in a case of bacterial meningitis caused by nontypeable H. influenzae infection in a child immunized with Hib vaccine.

  12. Meningitis y artritis por Haemophilus influenzae en un adulto

    Directory of Open Access Journals (Sweden)

    Javier Molina

    1988-02-01

    Full Text Available Tradicionalmente el Haemophilus influenzae ha sido considerado un germen causante de infecciones en niños; en adultos se lo ha relacionado con Infecciones respiratorias, pero en los últimos tiempos se han descrito en ellos infecciones severas cuando hay algunos factores predisponentes. Se describe un paciente drogadicto de 30 años con cuadro de meningitis y artritis y prueba de látex y cultivo de LCR positivos para HaemophiIus influenzae, quien recibió tratamiento con ampicilina, 2 gramos Intravenosos cada 4 horas y evolucionó a la mejoría sin secuelas. Se plantea la necesidad de tener en cuenta al Haemophilus influenzae como patógeno del adulto y más en aquellas personas con factores predisponentes.

    Haemophilus influenzae has traditionally been considered as an infectious agent that predominantly affects children; instead, in adults It has been Linked either to respiratory infections or to gevere infections occurring when predisposing factors are present. We describe a 30 year-old drug adict patient that presented with meningitis and arthritis; both latex test and cerebrospinal fluid culture were positive for Haemophilus influenzae. He was treated with ampicilin 2 gm, I. V. every four hours and improved without sequelae. This microorganism must be considered among those affecting adult patients specially when predisposing factors for infection are present.

  13. Probable acute disseminated encephalomyelitis due to Haemophilus influenzae meningitis

    OpenAIRE

    Beleza, P; M. Ribeiro; Pereira, J.; Jordão, MJ; Almeida, F.

    2008-01-01

    We report the case of a 17-year-old male on long-term steroid therapy for minimal lesion glomerulopathy who, after an upper respiratory infection, presented with Haemophilus influenzae type b meningitis. Twenty-four hours later he developed depression of consciousness which progressed to coma and left hemiparesis. Brain magnetic resonance imaging (MRI) revealed multiple lesions (hyperintense on T2 and slightly hypointense on Tl) involving mainly white matter suggestive of inflammation. MRI fe...

  14. Inducible repair system in Haemophilus influenzae unaccompanied by mutation. [uv

    Energy Technology Data Exchange (ETDEWEB)

    Notani, N.K.; Setlow, J.K.

    1980-07-01

    Weigle reactivation of ultraviolet-irradiated HPlc1 phage was observed after ultraviolet or mitomycin C treatment of Haemophilus influenzae cells. The amount of reactivation was considerably increased when the treated cells were incubated in growth medium before infection. The presence of chloramphenicol during this incubation abolished the reactivation. No mutation of this phage accompanied the reactivation. When cells were treated so as to produce a maximal reactivation of phage, neither reactivation nor mutation of cells was observed. It is concluded that H. influenzae has an inducible repair system that is not accompanied by mutation.

  15. Prevalence of genetic differences in phosphorylcholine expression between nontypeable Haemophilus influenzae and Haemophilus haemolyticus

    Directory of Open Access Journals (Sweden)

    Marrs Carl F

    2010-11-01

    Full Text Available Abstract Background Although non-typeable (NT Haemophilus influenzae and Haemophilus haemolyticus are closely related human commensals, H. haemolyticus is non-pathogenic while NT H. influenzae is an important cause of respiratory tract infections. Phase-variable phosphorylcholine (ChoP modification of lipooligosaccharide (LOS is a NT H. influenzae virulence factor that, paradoxically, may also promote complement activation by binding C-reactive protein (CRP. CRP is known to bind more to ChoP positioned distally than proximally in LOS, and the position of ChoP within LOS is dictated by specific licD alleles (designated here as licDI, licDIII, and licDIV that are present in a lic1 locus. The lic1 locus contains the licA-licD genes, and ChoP-host interactions may also be influenced by a second lic1 locus that allows for dual ChoP substitutions in the same strain, or by the number of licA gene tetranucleotide repeats (5'-CAAT-3' that reflect phase-variation mutation rates. Results Using dot-blot hybridization, 92% of 88 NT H. influenzae and 42.6% of 109 H. haemolyticus strains possessed a lic1 locus. Eight percent of NT H. influenzae and none of the H. haemolyticus strains possessed dual copies of lic1. The licDIII and licDIV gene alleles were distributed similarly (18-22% among the NT H. influenzae and H. haemolyticus strains while licDI alleles were present in 45.5% of NT H. influenzae but in less than 1% of H. haemolyticus strains (P H. influenzae had an average of 26.8 tetranucleotide repeats in licA compared to14.8 repeats in H. haemolyticus (P H. influenzae strains that possessed a licDIII allele had increased numbers of repeats compared to NT H. influenzae with other licD alleles (P Conclusions These data demonstrate that genetic similarities and differences of ChoP expression exist between NT H. influenzae and H. haemolyticus and strengthen the hypothesis that, at the population level, these differences may, in part, provide an advantage in

  16. Low occurrence of 'non-haemolytic Haemophilus haemolyticus' misidentified as Haemophilus influenzae in cystic fibrosis respiratory specimens, and frequent recurrence of persistent H. influenzae clones despite antimicrobial treatment.

    Science.gov (United States)

    Fenger, Mette G; Ridderberg, Winnie; Olesen, Hanne V; Nørskov-Lauritsen, Niels

    2012-12-01

    Non-influenzae commensal Haemophilus species of low pathogenicity may be difficult to discriminate from Haemophilus influenzae. We investigated the level of misidentifications in respiratory specimens from cystic fibrosis patients and evaluated the colonisation dynamics of genuine H. influenzae isolates. One hundred and ninety-two presumptive H. influenzae isolates were re-examined by assessment of marker genes sodC and fucK, and isolates with aberrant genotypes were subjected to multilocus sequence typing. Misidentifications (3%) were mainly caused by failure to identify porphyrin-synthesising strains, and only a single strain (0.5%) could be classified as 'non-haemolytic Haemophilus haemolyticus'. Sequential isolates of confirmed H. influenzae isolates from individual patients were typed by pulsed-field gel electrophoresis. Despite the routine prescription of antimicrobial therapy, the majority of H. influenzae isolates were identical with at least one of the strains cultured from the two preceding positive samples from the same patient. PMID:23177564

  17. A comparative study of preservation and storage of Haemophilus influenzae

    Directory of Open Access Journals (Sweden)

    Olga C Aulet de Saab

    2001-05-01

    Full Text Available The aim of this study was to compare the efficacy of conservation by freezing the strains of Haemophilus influenzae at -20ºC and -70ºC. Skim milk supplemented with glucose, yeast extract and glycerol allowed highest viability of H. influenzae both at -20ºC and -70ºC from the media analyzed. Trypticase soy broth and brain heart infusion broth supplemented with glycerol, allowed excellent recovery. Use of cotton swaps as supporting material, with or without addition of cryoprotective agents, did not modify H. influenzae viability after six months of storage. Concentration of the initial inoculum positively affected viability when stored at -20ºC. Initial concentration did not influence survival after storage at -70ºC. Thawing at room temperature should not exceed 3 h as to get highest survival percentage.

  18. Transformation of Haemophilus influenzae by plasmid RSF0885

    Energy Technology Data Exchange (ETDEWEB)

    Notani, N.K.; Setlow, J.K.; McCarthy, D.; Clayton, N.L.

    1981-12-01

    Plasmid RSF0885, which conferred ampicillin resistance, transformed competent Haemophilus influenzae cells with low efficiency (maximun, less than 0.01%). As judged by competition experiments and uptake of radioactivity, plasmid RSF0885 deoxyribonucleic acid was taken up into competent H. influenzae cells several orders of magnitude less efficiently than H. influenzae chromosomal deoxyribonucleic acid. Plasmid RSF0885 transformed cells with even lower efficiency than could be accounted for by the low uptake. Transformation was not affected by rec-1 and rec-2 mutations in the recipient, and strains cured of the plasmid did not show increased transformation. Plasmid molecules cut once with a restriction enzyme that made blunt ends did not transform. Transformation was favored by the closed circular form of the plasmid.

  19. Airway dysbiosis: Haemophilus influenzae and Tropheryma in poorly controlled asthma.

    Science.gov (United States)

    Simpson, Jodie L; Daly, Joshua; Baines, Katherine J; Yang, Ian A; Upham, John W; Reynolds, Paul N; Hodge, Sandra; James, Alan L; Hugenholtz, Philip; Willner, Dana; Gibson, Peter G

    2016-03-01

    Asthma is a chronic inflammatory disorder of the airways where bacteria may act as protagonists of chronic inflammation. Little is known about the relation of airway inflammation to the presence of specific bacterial taxa. We sought to describe the sputum microbiome in adults with poorly controlled asthma.DNA was extracted from induced sputum and microbial communities were profiled using 16S rRNA pyrosequencing. Bacterial species were characterised, and the relationship between microbial populations, asthma inflammatory subtypes and other covariates was explored. Real-time PCR was used to identify Tropheryma whipplei and Haemophilus influenzae in sputum.Adults with neutrophilic asthma had reduced bacterial diversity and species richness. Tropheryma was identified and confirmed with real-time PCR in 12 (40%) participants. Haemophilus occurred most often in a group of younger atopic males with an increased proportion of neutrophils. PCR confirmed the presence of H. influenzae in 35 (76%) participants with poorly controlled asthma.There are phenotype-specific alterations to the airway microbiome in asthma. Reduced bacterial diversity combined with a high prevalence of H. influenzae was observed in neutrophilic asthma, whereas eosinophilic asthma had abundant T. whipplei.

  20. Delineation of the species Haemophilus influenzae by phenotype, multilocus sequence phylogeny, and detection of marker genes

    DEFF Research Database (Denmark)

    Nørskov-Lauritsen, Niels; Overballe, MD; Kilian, Mogens

    2009-01-01

    To obtain more information on the much-debated definition of prokaryotic species, we investigated the borders of Haemophilus influenzae by comparative analysis of H. influenzae reference strains with closely related bacteria including strains assigned to Haemophilus haemolyticus, cryptic...... genospecies biotype IV, and the never formally validated species "Haemophilus intermedius". Multilocus sequence phylogeny based on six housekeeping genes separated a cluster encompassing the type and the reference strains of H. influenzae from 31 more distantly related strains. Comparison of 16S rRNA gene...... branching cluster, intermingled with strains of "H. intermedius" and cryptic genospecies biotype IV. Although H. influenzae is phenotypically more homogenous than some other Haemophilus species, the genetic diversity and multicluster structure of strains traditionally associated with H. influenzae make...

  1. Recurrent Posttraumatic Meningitis due to Nontypable Haemophilus influenzae: Case Report and Review of the Literature

    DEFF Research Database (Denmark)

    Kunze, W; Müller, L; Kilian, Mogens;

    2007-01-01

    We report a case of relapsing Haemophilus influenzae meningitis in a boy at the age of nearly 3 years and 4.2 years who had been successfully vaccinated against H. influenzae serotype b (Hib). The pathogen was a nonencapsulated (nontypable) H. influenzae strain of biotypes III and VI, respectively...

  2. Epidemiology of Haemophilus influenzae bacteremia: A multi-national population-based assessment

    DEFF Research Database (Denmark)

    Laupland, Kevin B; Schønheyder, Henrik C; Østergaard, Christian;

    2011-01-01

    OBJECTIVES: Haemophilus influenzae is an important cause of invasive infection but contemporary data in non-selected populations is limited. METHODS: Population-based surveillance for Haemophilus influenzae bacteremia was conducted in seven regions in Australia, Canada, and Denmark during 2000...... independently associated with death at 30-days in logistic regression analysis included male gender, hospital-onset disease, older age, and lower respiratory tract, central nervous system, or unknown focus of infection. CONCLUSIONS: Haemophilus influenzae is an important cause of morbidity and mortality......-2008. RESULTS: The overall annual incidence rate was 1.31 per 100,000 population and type specific rates were 0.08 for H. influenzae serotype b (Hib), 0.22 for H. influenzae serotypes a, c-f (Hiac-f), and 0.98 per 100,000 for non-typeable H. influenzae (NTHi). Very young and old patients were at highest risk...

  3. Haemophilus Influenzae Type b (Hib) Vaccine: What You Need to Know

    Science.gov (United States)

    VACCINE INFORMATION STATEMENT Hib Vaccine ( Haemophilus Influenzae Type b) What You Need to Know Many Vaccine Information Statements are available in Spanish and other languages. See www. immunize. org/ vis ...

  4. Phosphorylcholine allows for evasion of bactericidal antibody by Haemophilus influenzae.

    Directory of Open Access Journals (Sweden)

    Sarah E Clark

    Full Text Available The human pathogen Haemophilus influenzae has the ability to quickly adapt to different host environments through phase variation of multiple structures on its lipooligosaccharide (LPS, including phosphorylcholine (ChoP. During colonization with H. influenzae, there is a selection for ChoP+ phase variants. In a murine model of nasopharyngeal colonization, this selection is lost in the absence of adaptive immunity. Based on previous data highlighting the importance of natural antibody in limiting H. influenzae colonization, the effect of ChoP expression on antibody binding and its bactericidal activity was investigated. Flow cytometric analysis revealed that ChoP+ phase variants had decreased binding of antibody to LPS epitopes compared to ChoP- phase variants. This difference in antibody binding correlated with increased survival of ChoP+ phase variants in the presence of antibody-dependent, complement-mediated killing. ChoP+ phase variants were also more resistant to trypsin digestion, suggesting a general effect on the physical properties of the outer membrane. Moreover, ChoP-mediated protection against antibody binding correlated with increased resilience of outer membrane integrity. Collectively, these data suggest that ChoP expression provides a selective advantage during colonization through ChoP-mediated effects on the accessibility of bactericidal antibody to the cell surface.

  5. Probable acute disseminated encephalomyelitis due to Haemophilus influenzae meningitis.

    Science.gov (United States)

    Beleza, Pedro; Ribeiro, Manuel; Pereira, João; Ferreira, Carla; Jordão, Maria José; Almeida, Fátima

    2008-05-01

    We report the case of a 17-year-old male on long-term steroid therapy for minimal lesion glomerulopathy who, after an upper respiratory infection, presented with Haemophilus influenzae type b meningitis. Twenty-four hours later he developed depression of consciousness which progressed to coma and left hemiparesis. Brain magnetic resonance imaging (MRI) revealed multiple lesions (hyperintense on T2 and slightly hypointense on Tl) involving mainly white matter suggestive of inflammation. MRI features were compatible with acute disseminated encephalomyelitis (ADEM), although a differential diagnosis included cerebritis or vasculitis, secondary to bacterial meningitis. The patient was treated with high-dose steroids which resulted in a gradual improvement followed by complete clinical recovery. We propose a diagnosis of ADEM was the best diagnosis because of the radiological features and response to steroids. The occurrence of ADEM associated with acute meningitis, however rare, represents an important diagnostic challenge for the clinician. PMID:18355336

  6. Prophage induction and inactivation by uv light. [Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Barnhart, B.J.; Cox, S.H.; Jett, J.H.

    1976-06-01

    Analysis of the induction curves for uv light-irradiated Haemophilus influenzae lysogens and the distribution of pyrimidine dimers in a repair-deficient lysogen suggests that one dimer per prophage-size segment of the host bacterial chromosome is necessary as a preinduction event. The close correlations obtained prompted a renewed consideration of the possibility that direct prophage induction occurs when one dimer is stabilized within the prophage genome. The host excision-repair system apparently functions to reduce the probability of stabilizing within the prophage those dimers that are necessary for induction and inactivation. The presence of the inducible defective prophage in strain Rd depresses the inducibility of prophage HP1c1.

  7. Is Haemophilus influenzae better satellite for Enterococcus faecalis?

    Directory of Open Access Journals (Sweden)

    Firat Zafer Mengeloglu

    2013-09-01

    Full Text Available Background — Haemophilus influenzae can grow on blood agar media with Staphylococcus aureus which can provide factor V as it is called “Satellite phenomenon”. Objectives — In this study we tested and compared three different beta-haemolytic genus including three Staphylococcus aureus, three coagulase-negative staphylococci, and two Enterococcus faecalis strains in order to determine an alternative microorganism to be used for satellite test to identify H. influenzae conventionally. Materials and Methods — We used suspensions of H. influenzae in two different tribudities as 0.5 and 4 McFarland for each strain. Five totally-blinded reviewers examined the test results and scored both the colony sizes of H. influenzae and the diameter of the growth-zone. The sum of the scores for the colony sizes and the growth-zones were determined as “total diagnostic score” (TDS as being between 0-6 points for each test. Results — A total of 320 test scores were analysed. The mean TDS of E. faecalis group was significantly higher than the other groups (p<0.001. In the S. aureus group, 23 (19.2% tests had 0 points as TDS; but in enterococci group no isolates had lower scores than 3 points. In enterococci group, the rate of isolates which had 5 or 6 points was 77.5% (62/80; but in S. aureus group no isolate had higher than 4 points. Conclusions — Our study shows that using a beta-haemolytic E. faecalis strain will provide significantly more accurate results and will significantly reduce false-negative results for satellite test instead of S. aureus, which is particularly proposed to be used.

  8. Selective media for recovery of Haemophilus influenzae from specimens contaminated with upper respiratory tract microbial flora.

    OpenAIRE

    Chapin, K. C.; Doern, G V

    1983-01-01

    Isolation of Haemophilus influenzae from specimens contaminated with upper respiratory tract microbial flora was attempted with three different media: enriched chocolate agar, chocolate agar plus vancomycin, and chocolate agar plus vancomycin, bacitracin, and clindamycin. Recovery rates of H. influenzae from 852 pediatric pharyngeal swab specimens were 6.0, 28.5, and 59.9%, respectively.

  9. Immunogenicity and Efficacy of Different Haemophilus influenzae type b Vaccines

    Directory of Open Access Journals (Sweden)

    Mojgani, N.

    2014-11-01

    Full Text Available Haemophilus influenzae, a major cause of meningitis in young children leading to death and other neurological sequelae. The disease leaves 15 to 35% of the survivors with permanent disabilities, such as, mental retardation or deafness. Despite the availability of new and more powerful antibiotics children with Hib meningitis still suffer from high mortality or morbidity. The emergence of multiresistant Hib strains causes increasing difficulties in selecting proper antibiotics for the treatment. Since 1970, the capsular polysaccharide polyribosylribitol phosphate (PRP in H. influenzae b has been the target for vaccine development. The first Hib polysaccharide vaccine licensed in 1985, proved immunogenic in human adults, but failed to elicit an immune response in children under 2 years of age who were at greatest risk of developing the invasive Hib infection. These factors led to one of the most exciting advances in pediatrics, the development of Hib conjugate vaccines. Unlike most other vaccines for preventing a particular disease which are generally similar for all types, the specific characteristics of the available Hib conjugate vaccines licensed vary from each other in structure and immunological properties. In this review the immunogenicity and efficacy of Hib vaccines including a PRP vaccine; b Conjugate vaccines; and c Combination vaccines is evaluated.

  10. Molecular Surveillance of True Nontypeable Haemophilus influenzae: An Evaluation of PCR Screening Assays

    OpenAIRE

    Binks, Michael J; Beth Temple; Lea-Ann Kirkham; Wiertsema, Selma P; Dunne, Eileen M.; Richmond, Peter C.; Marsh, Robyn L; Leach, Amanda J; Smith-Vaughan, Heidi C

    2012-01-01

    BACKGROUND: Unambiguous identification of nontypeable Haemophilus influenzae (NTHi) is not possible by conventional microbiology. Molecular characterisation of phenotypically defined NTHi isolates suggests that up to 40% are Haemophilus haemolyticus (Hh); however, the genetic similarity of NTHi and Hh limits the power of simple molecular techniques such as PCR for species discrimination. METHODOLOGY/PRINCIPAL FINDINGS: Here we assess the ability of previously published and novel PCR-based ass...

  11. Biofilm-specific extracellular matrix proteins of nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Wu, Siva; Baum, Marc M; Kerwin, James; Guerrero, Debbie; Webster, Simon; Schaudinn, Christoph; VanderVelde, David; Webster, Paul

    2014-12-01

    Nontypeable Haemophilus influenzae (NTHi), a human respiratory tract pathogen, can form colony biofilms in vitro. Bacterial cells and the amorphous extracellular matrix (ECM) constituting the biofilm can be separated using sonication. The ECM from 24- and 96-h NTHi biofilms contained polysaccharides and proteinaceous components as detected by nuclear magnetic resonance (NMR) and Fourier transform infrared spectroscopy (FTIR) spectroscopy. More conventional chemical assays on the biofilm ECM confirmed the presence of these components and also DNA. Proteomics revealed eighteen proteins present in biofilm ECM that were not detected in planktonic bacteria. One ECM protein was unique to 24-h biofilms, two were found only in 96-h biofilms, and fifteen were present in the ECM of both 24- and 96-h NTHi biofilms. All proteins identified were either associated with bacterial membranes or cytoplasmic proteins. Immunocytochemistry showed two of the identified proteins, a DNA-directed RNA polymerase and the outer membrane protein OMP P2, associated with bacteria and biofilm ECM. Identification of biofilm-specific proteins present in immature biofilms is an important step in understanding the in vitro process of NTHi biofilm formation. The presence of a cytoplasmic protein and a membrane protein in the biofilm ECM of immature NTHi biofilms suggests that bacterial cell lysis may be a feature of early biofilm formation.

  12. Mutation induction by MNNG in a bacteriophage of Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Boling, M.E.; Kimball, R.F.

    1976-01-01

    Three temperature-sensitive mutants of the haemophilus influenza phage HP1c1 were tested for reversion to wild type (ts ..-->.. ts/sup +/). Treatment with N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) produced revertants at levels up to 0.1% of the total progeny phage from treated lysogens. Cells treated with MNNG after infection with whole ts phage produced progeny phage with similar reversion frequencies, but when the uninfected cells or the phage were treated alone no reversion was induced. Fixation of premutational lesions was shown to occur with no evidence for host-cell DNA synthesis, indicating that phage DNA synthesis may be responsible for fixation of mutation in phage DNA. Evidence is given which shows that prophage DNA replicating by the cells' replicating system after treatment and before induction, produces the same number of revertants per survivor as phage DNA which is replicated outside the host genome. Two of the phage mutants (ts1 and ts2) reverted at similar frequencies, while one of the mutants (ts3) exhibited a much lower induced reversion frequency.

  13. The Role of Cytomegalovirus, Haemophilus Influenzae and Epstein Barr Virus in Guillain Barre Syndrome

    Directory of Open Access Journals (Sweden)

    Shahriar Nafissi

    2013-06-01

    Full Text Available Guillain Barre Syndrome (GBS is an inflammatory, usually demyelinating, polyneuropathy; clinically characterized by acute onset of symmetric progressive muscle weakness with loss of myotatic reflexes. Thirty five patients with GBS, defined clinically according to the criteria of Asbury and Cornblath, were recruited from three hospital affiliated to Tehran University of Medical Sciences. Controls: As a control group 35 age and sex matched patients with other neurological diseases admitted to the same hospital at the same time, were included in our study. Serum samples were collected before treatment from each patient (within 4 weeks after the disease onset and controls, and stored frozen at -80ºC until serologic assays were done. Serologic testing of pretreatment serum was performed in all patients. Positive titer of virus specific IgM antibody against cytomegalovirus (CMV was found in 6 cases and 2 controls. 34 patients and 31 controls had high titer of anti Haemophilus influenzae IgG and one patient had serologic evidence of a recent Epstein Barr virus (EBV infection. The mean titer of IgG antibody against Haemophilus influenzae in cases and controls was 5.21 and 2.97 respectively. Although serologic evidence of all these infections were more frequent in cases than in controls, only Haemophilus influenzae infection appeared to be significantly related to GBS (P=0.002. Eleven cases and 3 controls had high titers of IgG antibody against Haemophilus influenzae type B (titer >8. There is significant association between high titer of IgG antibody against Haemophilus influenzae and GBS (P=0.017. Our results provide further evidence that Haemophilus influenzae and probably CMV, can be associated with GBS

  14. The role of cytomegalovirus, Haemophilus influenzae and Epstein Barr virus in Guillain Barre syndrome.

    Directory of Open Access Journals (Sweden)

    Shahriar Nafissi

    2013-06-01

    Full Text Available Guillain Barre Syndrome (GBS is an inflammatory, usually demyelinating, polyneuropathy; clinically characterized by acute onset of symmetric progressive muscle weakness with loss of myotatic reflexes. Thirty five patients with GBS, defined clinically according to the criteria of Asbury and Cornblath, were recruited from three hospital affiliated to Tehran University of Medical Sciences.As a control group 35 age and sex matched patients with other neurological diseases admitted to the same hospital at the same time, were included in our study. Serum samples were collected before treatment from each patient (within 4 weeks after the disease onset and controls, and stored frozen at -80ºC until serologic assays were done. Serologic testing of pretreatment serum was performed in all patients. Positive titer of virus specific IgM antibody against cytomegalovirus (CMV was found in 6 cases and 2 controls. 34 patients and 31 controls had high titer of anti Haemophilus influenzae IgG and one patient had serologic evidence of a recent Epstein Barr virus (EBV infection. The mean titer of IgG antibody against Haemophilus influenzae in cases and controls was 5.21 and 2.97 respectively. Although serologic evidence of all these infections were more frequent in cases than in controls, only Haemophilus influenzae infection appeared to be significantly related to GBS (P=0.002. Eleven cases and 3 controls had high titers of IgG antibody against Haemophilus influenzae type B (titer >8. There is significant association between high titer of IgG antibody against Haemophilus influenzae and GBS (P=0.017. Our results provide further evidence that Haemophilus influenzae and probably CMV, can be associated with GBS.

  15. Occurrence of Haemophilus influenzae strains in three Brazilian states since the introduction of a conjugate Haemophilus influenzae type b vaccine

    Directory of Open Access Journals (Sweden)

    de Almeida A.E.C.C.

    2005-01-01

    Full Text Available Few vaccines in history have induced such a dramatic decline in incidence over such a short period of time as the Haemophilus influenzae type b (Hib conjugate. This vaccine was introduced in 1988 in the United States, but only in 1999 was Hib immunization introduced by the Brazilian Ministry of Health as part of the routine infant National Immunization Program. The authors analyzed 229 H. influenzae (Hi isolates from Public Health Laboratories in three Brazilian states: Pernambuco (Northeast, N = 54, Santa Catarina (South, N = 19, and Rio de Janeiro (Southeast, N = 156. The isolates were collected from Brazilian children 0-10 years of age with meningitis and other infections from 1990 to 2003 and were part of the research collection of the National Institute of Quality Control in Health, FIOCRUZ. Bacterial strains were characterized by serotyping and biotyping. During the pre-vaccination period the prevalence infection due to Hib was of 165 isolates and only 2 non-b Hi among all the notified meningitis infections caused by Hi. Our results showed a significant decrease in the prevalence of Hib meningitis from 165 to 33 isolates after 1999. However, during the post-vaccination period of 2001-2003 we observed an increase in the number of non-b Hi isolates: only 2 non-b strains isolated from 1990 to 1999 and 29 from 1999 to 2003. Based on the present data, the authors emphasize the need for more sensitive epidemiological and bacteriological studies aiming the improvement of the available Hib vaccine, in order to protect the susceptible population to infections due to other serological types of Hi and the reevaluation of immunization schedules used by the National Immunization Program.

  16. Electroporation of Haemophilus influenzae is effective for transformation of plasmid but not chromosomal DNA.

    OpenAIRE

    Mitchell, M A; Skowronek, K.; Kauc, L; Goodgal, S H

    1991-01-01

    Electroporation of plasmid and chromosomal DNAs were tested in Haemophilus influenzae because of an interest in introducing DNA into mutants that are deficient in competence for transformation. The initial experiments were designed to investigate and optimize conditions for electroporation of H. influenzae. Plasmid DNA was introduced into the competence proficient strain Rd and its competence-deficient uptake mutants com-52, com-59, and com-88, and the recombination deficient mutant rec1. Pla...

  17. Haemophilus influenzae Infection Drives IL-17-Mediated Neutrophilic Allergic Airways Disease

    OpenAIRE

    Essilfie, Ama-Tawiah; Simpson, Jodie L.; Horvat, Jay C.; Julie A Preston; Dunkley, Margaret L.; Paul S Foster; Gibson, Peter G; Hansbro, Philip M

    2011-01-01

    A subset of patients with stable asthma has prominent neutrophilic and reduced eosinophilic inflammation, which is associated with attenuated airways hyper-responsiveness (AHR). Haemophilus influenzae has been isolated from the airways of neutrophilic asthmatics; however, the nature of the association between infection and the development of neutrophilic asthma is not understood. Our aim was to investigate the effects of H. influenzae respiratory infection on the development of hallmark featu...

  18. Haemophilus influenzae infection drives IL-17-mediated neutrophilic allergic airways disease.

    OpenAIRE

    Ama-Tawiah Essilfie; Simpson, Jodie L.; Horvat, Jay C.; Julie A Preston; Dunkley, Margaret L.; Paul S Foster; Gibson, Peter G; Hansbro, Philip M

    2011-01-01

    A subset of patients with stable asthma has prominent neutrophilic and reduced eosinophilic inflammation, which is associated with attenuated airways hyper-responsiveness (AHR). Haemophilus influenzae has been isolated from the airways of neutrophilic asthmatics; however, the nature of the association between infection and the development of neutrophilic asthma is not understood. Our aim was to investigate the effects of H. influenzae respiratory infection on the development of hallmark featu...

  19. Genetic Characterization of the Capsulation Locus of Haemophilus influenzae Serotype e▿

    OpenAIRE

    Giufrè, Maria; Cardines, Rita; Mastrantonio, Paola; Cerquetti, Marina

    2010-01-01

    The capsulation (cap) locus of Haemophilus influenzae type e (Hie) was characterized and sequenced. No IS1016 element was found to flank the locus. The 18.2-kb locus included 14 open reading frames (ORFs), which were grouped into three functional regions. Eight new ORFs (named ecs1 to ecs8) were identified in the Hie capsule-specific region II.

  20. Activity of ciprofloxacin and azithromycin on biofilms produced in vitro by Haemophilus influenzae

    Institute of Scientific and Technical Information of China (English)

    WANG Dong; WANG Ying; LIU You-ning

    2009-01-01

    Background It is recognized that Haemophilus influenzae isolated from patients with otitis media forms biofilms both in vitro and in vivo, suggesting that biofilm formation in vivo might play an important role in the pathogenesis and chronicity of otitis media, but the effect of antibiotics on biofilm has not been well studied. We investigated the impact of ciprofloxacin and azithromycin on bacterial biofilms formed by Haemophilus influenzae in vitro in this study.Methods Eleven strains of Haemophilus influenzae were isolated from sputum specimens collected from patients with acute exacerbation of chronic obstructive pulmonary diseases. Formation of bacterial biofilm was examined by crystal violet assay and a scanning electron microscope. Alterations of biofilms were measured under varying concentrations of azithromycin and ciprofloxacin.Results Striking differences were observed among strains with regard to the ability to form biofilm. Typical membrane-like structure formed by bacterial cells and extracellular matrix was detected. Initial biofilm synthesis was inhibited by azithromycin and ciprofloxacin at concentrations higher than two-fold minimal inhibitory concentration.Disruption of mature biofilms could be achieved at relatively higher concentration, and ciprofloxacin displayed more powerful activity.Conclusions Haemophilus influenzae is capable of forming biofilm in vitro. Sufficient dosage might control early formation of biofilms. Ciprofloxacin exerts better effects on breakdown of biofilm than azithromycin at conventional concentration in clinics.

  1. A REVIEW OF THERAPEUTIC MONITORING OF CHLORAMPHENICOL IN PATIENTS WITH HAEMOPHILUS-INFLUENZAE MENINGITIS

    NARCIS (Netherlands)

    COAKLEY, JC; HUDSON, [No Value; SHANN, F; CONNELLY, JF

    1992-01-01

    Two hundred and seventy-seven serum chloramphenicol concentrations in 90 patients with Haemophilus influenzae type b meningitis were analysed retrospectively- Most patients were given chloramphenicol 25 mg/kg 6 hourly initially. Chloramphenicol concentrations were categorized as pre-dose ('trough')

  2. Structural requirements of the major protective antibody to Haemophilus influenzae type b

    DEFF Research Database (Denmark)

    Hougs, L; Juul, L; Svejgaard, A;

    1999-01-01

    Protective antibodies to the important childhood pathogen Haemophilus influenzae type b (Hib) are directed against the capsular polysaccharide (HibCP). Most of the antibody is encoded by a well-defined set of ("canonical") immunoglobulin genes, including the Vkappa A2 gene, and expresses...

  3. Evaluation of Haemophilus influenzae Type B Conjugate Vaccine (Meningococcal Protein Conjugate in Canadian Infants

    Directory of Open Access Journals (Sweden)

    David W Scheifele

    1994-01-01

    Full Text Available Objective: To assess adverse effects and immune responses with a three-dose series of Haemophilus influenzae type b meningococcal protein conjugate (PedvaxHIB or Hib.OMP vaccine, including any immunological response alterations from concurrent administration with routine vaccines for infants.

  4. Antibody response to Haemophilus influenzae type b capsular polysaccharide conjugated to tetanus toxoid in preterm infants

    DEFF Research Database (Denmark)

    Kristensen, Kim; Gyhrs, A; Lausen, B;

    1996-01-01

    OBJECTIVE: To evaluate the antibody response to a Haemophilus influenzae type b capsular polysaccharide (HibCP) tetanus toxoid (TT) conjugate vaccine (HibCP-TT) in preterm infants. SUBJECTS: Thirty-five healthy preterm infants with gestational ages (GA) from 27 to 36 weeks and birth weights from...

  5. Analysis of Haemophilus influenzae serotype f isolated from three Japanese children with invasive H. influenzae infection.

    Science.gov (United States)

    Hoshino, Tadashi; Hachisu, Yushi; Kikuchi, Takashi; Tokutake, Shoko; Okui, Hideyuki; Kutsuna, Satoru; Fukasawa, Chie; Murayama, Kei; Oohara, Asami; Shimizu, Hiroyuki; Ito, Midori; Takahashi, Yoshiko; Ishiwada, Naruhiko

    2015-04-01

    In Japan, publicly subsidized Haemophilus influenzae serotype b vaccines became available in 2011; consequently, the incidence of invasive H. influenzae infection in paediatric patients of less than 5 years of age decreased dramatically. In 2013, the first case of H. influenzae serotype f (Hif) meningitis in a Japanese infant was reported, and another case of Hif meningitis in a Japanese infant was observed in 2013. We experienced a fatal paediatric case of Hif bacteraemia in 2004; therefore, we conducted an analysis of the three Hif strains isolated from these three Japanese children with invasive Hif infections. All three strains were β-lactamase-non-producing, ampicillin-sensitive strains, with MICs of 1 µg ml(-1) or less. However, one of the three strains showed slightly elevated MICs for ampicillin (1 µg ml(-1)), cefotaxime (0.25 µg ml(-1)) and meropenem (0.13 µg ml(-1)). A molecular analysis by multilocus sequence typing identified all three strains as sequence type (ST) 124, which is a predominant invasive Hif strain in many countries. SmaI-digested PFGE showed variable DNA fragmentation patterns among the strains, suggesting that some highly virulent strains have originated from a single ST124 clone and caused invasive Hif infections in Japan. Additional studies are needed to determine the factors that have led to the clonal expansion of virulent ST124 strains.

  6. Intra-tracheal Administration of Haemophilus influenzae in Mouse Models to Study Airway Inflammation.

    Science.gov (United States)

    Venuprasad, K; Theivanthiran, Balamayooran; Cantarel, Brandi

    2016-03-02

    Here, we describe a detailed procedure to efficiently and directly deliver Haemophilus influenzae into the lower respiratory tracts of mice. We demonstrate the procedure for preparing H. influenzae inoculum, intra-tracheal instillation of H. influenzae into the lung, collection of broncho-alveolar lavage fluid (BALF), analysis of immune cells in the BALF, and RNA isolation for differential gene expression analysis. This procedure can be used to study the lung inflammatory response to any bacteria, virus or fungi. Direct tracheal instillation is mostly preferred over intranasal or aerosol inhalation procedures because it more efficiently delivers the bacterial inoculum into the lower respiratory tract with less ambiguity.

  7. Simultaneous Specific Detection of Streptococcus pneumoniae, Haemophilus influenzae and Mycoplasma pneumoniae in Sputum Samples from Patients with Suspected Influenza by Multiplex-PCR

    Directory of Open Access Journals (Sweden)

    Amin Moazami

    2014-01-01

    Full Text Available Abstract Background and objective: Streptococcus pneumoniae, Haemophilus influenza and Mycoplasma pneumoniae are the most common cause in bacterial pneumonia. Also these agents can cause bacterial superinfection in patients with influenza. Aim of this study was Simultaneous specific detection of Streptococcus pneumoniae, Haemophilus influenza and Mycoplasma pneumoniae in sputum samples from patients with suspected influenza by Multiplex-PCR. Materials and methods: In this study, 170 sputum samples in patients with suspected influenza with age from 3 months to 70 years, received the Influenza Reference Laboratory – Tehran Medical university were tested by Multiplex PCR. Amplified DNA fragments size was 394 bp for Streptococcus pneumoniae, 199 bp for Haemophilus influenzae and 416 bp for Mycoplasma pneumoniae. Results: of all 170 samples, 30 samples were positive for Streptococcus pneumoniae and Haemophilus influenza. Of the 30 positive samples, 27 samples (15/8 % and 3 samples (1/7 % were positive for S. pneumoniae, Haemophilus influenza respectively. Conclusion: This study showed that Multiplex-PCR able to diagnosis desired bacteria in short time and so this molecular method can use as complementary technique especially when the results of gram stain, culture or serological test are negative.

  8. Duplex Quantitative PCR Assay for Detection of Haemophilus influenzae That Distinguishes Fucose- and Protein D-Negative Strains.

    Science.gov (United States)

    de Gier, Camilla; Pickering, Janessa L; Richmond, Peter C; Thornton, Ruth B; Kirkham, Lea-Ann S

    2016-09-01

    We have developed a specific Haemophilus influenzae quantitative PCR (qPCR) that also identifies fucose-negative and protein D-negative strains. Analysis of 100 H. influenzae isolates, 28 Haemophilus haemolyticus isolates, and 14 other bacterial species revealed 100% sensitivity (95% confidence interval [CI], 96% to 100%) and 100% specificity (95% CI, 92% to 100%) for this assay. The evaluation of 80 clinical specimens demonstrated a strong correlation between semiquantitative culture and the qPCR (P < 0.001).

  9. Haemophilus haemolyticus Interaction with Host Cells Is Different to Nontypeable Haemophilus influenzae and Prevents NTHi Association with Epithelial Cells.

    Science.gov (United States)

    Pickering, Janessa L; Prosser, Amy; Corscadden, Karli J; de Gier, Camilla; Richmond, Peter C; Zhang, Guicheng; Thornton, Ruth B; Kirkham, Lea-Ann S

    2016-01-01

    Nontypeable Haemophilus influenzae (NTHi) is an opportunistic pathogen that resides in the upper respiratory tract and contributes to a significant burden of respiratory related diseases in children and adults. Haemophilus haemolyticus is a respiratory tract commensal that can be misidentified as NTHi due to high levels of genetic relatedness. There are reports of invasive disease from H. haemolyticus, which further blurs the species boundary with NTHi. To investigate differences in pathogenicity between these species, we optimized an in vitro epithelial cell model to compare the interaction of 10 H. haemolyticus strains with 4 NTHi and 4 H. influenzae-like haemophili. There was inter- and intra-species variability but overall, H. haemolyticus had reduced capacity to attach to and invade nasopharyngeal and bronchoalveolar epithelial cell lines (D562 and A549) within 3 h when compared with NTHi. H. haemolyticus was cytotoxic to both cell lines at 24 h, whereas NTHi was not. Nasopharyngeal epithelium challenged with some H. haemolyticus strains released high levels of inflammatory mediators IL-6 and IL-8, whereas NTHi did not elicit an inflammatory response despite higher levels of cell association and invasion. Furthermore, peripheral blood mononuclear cells stimulated with H. haemolyticus or NTHi released similar and high levels of IL-6, IL-8, IL-10, IL-1β, and TNFα when compared with unstimulated cells but only NTHi elicited an IFNγ response. Due to the relatedness of H. haemolyticus and NTHi, we hypothesized that H. haemolyticus may compete with NTHi for colonization of the respiratory tract. We observed that in vitro pre-treatment of epithelial cells with H. haemolyticus significantly reduced NTHi attachment, suggesting interference or competition between the two species is possible and warrants further investigation. In conclusion, H. haemolyticus interacts differently with host cells compared to NTHi, with different immunostimulatory and cytotoxic

  10. Multilocus Sequencing Typing of Invasive Haemophilus influenzae strains Isolated in Portugal in the Pre-vaccination Period (1989-2001)

    OpenAIRE

    Veiga, Elisabete; Gomes, Sandra; Bettencourt, Célia; Bajanca-Lavado, Maria Paula

    2015-01-01

    Introduction: Haemophilus influenzae can cause life-threatening infections in children and adults, such as pneumonia, bacteremia, and meningitis, despite de availability of the H. influenzae type b vaccine. Six capsular types, a-f, have been identified to date. Non-capsulated (NC) H. influenzae have also been described. Multilocus Sequencing Typing (MLST) is a powerful method that allows a precise and unambiguous characterization of H. influenzae genotypes. Aim: Identification of the maj...

  11. A review of the role of Haemophilus influenzae in community-acquired pneumonia

    Directory of Open Access Journals (Sweden)

    Mary PE Slack

    2015-06-01

    Full Text Available In an era when Haemophilus influenzae type b (Hib conjugate vaccine is widely used, the incidence of Hib as a cause of community-acquired pneumonia (CAP has dramatically declined. Non-typeable H. influenzae (NTHi strains and, occasionally, other encapsulated serotypes of H. influenzae are now the cause of the majority of invasive H. influenzae infections, including bacteraemic CAP. NTHi have long been recognised as an important cause of lower respiratory tract infection, including pneumonia, in adults, especially those with underlying diseases. The role of NTHi as a cause of non-bacteraemic CAP in children is less clear. In this review the evidence for the role of NTHi and capsulated strains of H. influenzae will be examined.

  12. Epidemiological Monitoring and Prevention of Haemophilus influenzae type b in the RF

    Directory of Open Access Journals (Sweden)

    E. Ya. Frolova

    2012-01-01

    Full Text Available At three study sites, incidence rates of meningitis caused by Haemophilus influenzae type b exceeded the threshold of 10 cases per на 100,000 children under 5 and mandated vaccination. Both vaccines the use of which is permitted in the Russian Federation are efficacious and safe. In order to make immunization a success, it’s imperative to take the comprehensive approach comprising not only vaccine procurement but also cold chain reinforcement, laboratory capacity building including staff training, computerization of primary care facilities as well as other feasible types of healthcare modernization. Immunization against Haemophilus influenzae type b not only prevents in children purulent bacterial meningitis of aforementioned etiology but also reduces rates of respiratory infections.

  13. Distribution and Diversity of hmw1A Among Invasive Nontypeable Haemophilus influenzae Isolates in Iran

    Science.gov (United States)

    Shahini Shams Abadi, Milad; Siadat, Seyed Davar; Vaziri, Farzam; Davari, Mehdi; Fateh, Abolfazl; Pourazar, Shahin; Abdolrahimi, Farid; Ghazanfari, Morteza

    2016-01-01

    Background: The pathogenesis of nontypeable Haemophilus influenzae (NTHi) begins with adhesion to the rhinopharyngeal mucosa. Almost 38–80% of NTHi clinical isolates produce proteins that belong to the High Molecular Weight (HMW) family of adhesins, which are believed to facilitate colonization. Methods: In the present study, the prevalence of hmwA, which encodes the HMW adhesin, was determined for a collection of 32 NTHi isolates. Restriction Fragment Length Polymorphism (RFLP) was performed to advance our understanding of hmwA binding sequence diversity. Results: The results demonstrated that hmwA was detected in 61% of NTHi isolates. According to RFLP, isolates were divided into three groups. Conclusion: Based on these observations, it is hypothesized that some strains of nontypeable Haemophilus influenzae infect some specific areas more than other parts. PMID:27141269

  14. Heme Utilization by Nontypeable Haemophilus influenzae Is Essential and Dependent on Sap Transporter Function▿†

    OpenAIRE

    Mason, Kevin M.; Raffel, Forrest K.; Ray, William C.; Bakaletz, Lauren O.

    2011-01-01

    Bacterial strategies of innate immune evasion and essential metabolic functions are critical for commensal-host homeostasis. Previously, we showed that Sap translocator function is necessary for nontypeable Haemophilus influenzae (NTHI) behaviors that mediate diseases of the human airway. Antimicrobial peptide (AP) lethality is limited by binding mediated by the Sap complex. SapA shares homology with the dipeptide-binding protein (DppA) and the heme-binding lipoprotein (HbpA), both of which h...

  15. Outer membrane protein binding sites of complement component 3 during opsonization of Haemophilus influenzae.

    OpenAIRE

    Hetherington, S V; Patrick, C C; Hansen, E J

    1993-01-01

    Complement component 3 (C3) binding to Haemophilus influenzae type b (Hib) is an important step in host defense against invasive disease, but the details of this process remain poorly understood. We have shown that the P1 and P2 outer membrane proteins (OMPs) serve as binding sites for C3 on serum-opsonized Hib. Whole-cell lysates of opsonized Hib were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the resolved proteins were transferred to nitrocellulose. Immunobl...

  16. Invasive Haemophilus Influenzae Disease, Europe, 1996–2006

    Centers for Disease Control (CDC) Podcasts

    2010-03-15

    This podcast describes monitoring of Haemophilus influenzae disease in Europe from 1996 through 2006. CDC epidemiologist Stacey Martin discusses what researchers learned about the effect of vaccination on disease prevalence.  Created: 3/15/2010 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID); National Center for Immunization and Respiratory Diseases (NCIRD).   Date Released: 4/5/2010.

  17. Molecular characterization of "plasmid-free" antibiotic-resistant Haemophilus influenzae.

    OpenAIRE

    Roberts, M C; Smith, A. L.

    1980-01-01

    We examined 14 multiresistant and 8 ampicillin- or tetracycline-resistant Haemophilus influenzae isolates and 4 ampicillin-resistant H. parainfluenzae isolates for plasmid deoxyribonucleic acid. Sixteen strains carried plasmids. Both "plasmid-free" and plasmid-carrying isolates transferred the antibiotic resistance by conjugation. All transconjugants carried plasmid deoxyribonucleic acid, suggesting that the apparent plasmid-free strains contained R plasmids encoding for antibiotic resistance.

  18. Functional characterization of UvrD helicases from Haemophilus influenzae and Helicobacter pylori.

    Science.gov (United States)

    Sharma, Ruchika; Rao, Desirazu N

    2012-06-01

    Haemophilus influenzae and Helicobacter pylori are major bacterial pathogens that face high levels of genotoxic stress within their host. UvrD, a ubiquitous bacterial helicase that plays important roles in multiple DNA metabolic pathways, is essential for genome stability and might, therefore, be crucial in bacterial physiology and pathogenesis. In this study, the functional characterization of UvrD helicase from Haemophilus influenzae and Helicobacter pylori is reported. UvrD from Haemophilus influenzae (HiUvrD) and Helicobacter pylori (HpUvrD) exhibit strong single-stranded DNA-specific ATPase and 3'-5' helicase activities. Mutation of highly conserved arginine (R288) in HiUvrD and glutamate (E206) in HpUvrD abrogated their activities. Both the proteins were able to bind and unwind a variety of DNA structures including duplexes with strand discontinuities and branches, three- and four-way junctions that underpin their role in DNA replication, repair and recombination. HiUvrD required a minimum of 12 nucleotides, whereas HpUvrD preferred 20 or more nucleotides of 3'-single-stranded DNA tail for efficient unwinding of duplex DNA. Interestingly, HpUvrD was able to hydrolyze and utilize GTP for its helicase activity although not as effectively as ATP, which has not been reported to date for UvrD characterized from other organisms. HiUvrD and HpUvrD were found to exist predominantly as monomers in solution together with multimeric forms. Noticeably, deletion of distal C-terminal 48 amino acid residues disrupted the oligomerization of HiUvrD, whereas deletion of 63 amino acids from C-terminus of HpUvrD had no effect on its oligomerization. This study presents the characteristic features and comparative analysis of Haemophilus influenzae and Helicobacter pylori UvrD, and constitutes the basis for understanding the role of UvrD in the biology and virulence of these pathogens. PMID:22500516

  19. Haemophilus influenzae Type b Carriage and Novel Bacterial Population Structure among Children in Urban Kathmandu, Nepal▿

    OpenAIRE

    Williams, E. J.; Lewis, J.; John, T.; Hoe, J. C.; Yu, L.; Dongol, S.; Kelly, D. F.; Griffiths, D. T.; Shah, A; Limbu, B.; Pradhan, R.; Mawas, F.; Shrestha, S.; Thorson, S.; Werno, A. M.

    2011-01-01

    Haemophilus influenzae type b (Hib) is a major cause of invasive bacterial infection in children that can be prevented by a vaccine, but there is still uncertainty about its relative importance in Asia. This study investigated the age-specific prevalence of Hib carriage and its molecular epidemiology in carriage and disease in Nepal. Oropharyngeal swabs were collected from children in Kathmandu, Nepal, from 3 different settings: a hospital outpatient department (OPD), schools, and children's ...

  20. sxy-1, a Haemophilus influenzae mutation causing greatly enhanced spontaneous competence.

    OpenAIRE

    Redfield, R J

    1991-01-01

    A Haemophilus influenzae strain carrying a competence-enhancing mutation (sxy-1) was selected by transformation of a mutagenized culture in exponential growth at low cell density, where spontaneous competence is very rare. Under these conditions, sxy-1 cells spontaneously transformed 100 to 1,000 times more efficiently than wild-type cells. Moreover, sxy-1 cells responded to all known competence-inducing treatments with further increases in transformation frequency. At high cell densities, sx...

  1. VAKSIN HAEMOPHILUS INFLUENZAE type b (Hib UNTUK PENCEGAHAN MENINGITIS DAN PNEUMONIA

    Directory of Open Access Journals (Sweden)

    Muljati Prijanto

    2012-09-01

    Full Text Available Rendahnya insiden pada anak yang lebih tua umurnya dan orang dewasa disebabkan karena adanya antibodi bakterisidal terhadap capsul polisakarida Haemophilus influenzae tipe b dalam serumnya.Di negara maju maupun negara berkembang bakteri merupakan penyebab utama non epidemik meningitis pada kelompok umur tersebut. Beberapa bukti menunjukkan bahwa mortalitas meningitis pada kelompok bayi muda lebih tinggi di negara berkembang dari pada di negara industri. 

  2. Mutation induction in Haemophilus influenzae by ICR-191 II. Role of DNA replication and repair

    Energy Technology Data Exchange (ETDEWEB)

    Kimball, R.F.; Perdue, S.W.

    1981-01-01

    Evidence is presented to show that presumptive frameshift mutations induced in Haemophilus influenzae by ICR-191 are fixed very repidly, essentially at the time of treatment. DNA synthesis during treatment is essential for fixation, but DNA synthesis after treatment has no effect. The conclusion is drawn that the mutagen acts at the replication fork, possibly to stabilize misannealings arising in association with the discontinuities in the newly synthesized DNA. (JMT)

  3. Haemophilus influenzae as a cause of Brodie's abscess in an infant.

    Science.gov (United States)

    Kurlandsky, L E; Quinn, P H; Sills, E M

    1979-01-01

    Brodie's abscess is a form of subacute osteomyelitis which is defined by a particular constellation of clinical, radiological and pathological features. Its occurrence in infants is extremely rare. This case documents just such an occurrence. To our knowledge, the pathogen Haemophilus influenzae has not been previously recognized as a cause of Brodie's abscess in particular or subacute osteomyelitis in general. The clinical presentation and diagnostic pitfalls which may be encountered in this age group are discussed.

  4. Adult-onset Invasive Haemophilus influenzae Type f Caused by Acute Lower Leg Cellulitis.

    Science.gov (United States)

    Usui, Yuko; Kakuta, Risako; Araki, Makoto; Sato, Taigo; Gu, Yoshiaki; Yano, Hisakazu; Taniuchi, Norihide

    2016-01-01

    In Japan, routine Haemophilus influenzae type b (Hib) vaccination began in 2013. Thus, similar to other countries, a strain shift is expected in the near future. We experienced a case of H. influenzae type f (Hif) bacteremia in a 66-year-old man. The primary focus of the infection was the soft tissue of the left lower leg, which is an extremely rare origin in adults. Subsequently, we conducted multilocus sequence typing and identified the strain as sequence type 124, which is the most common invasive strain of Hif worldwide. This case may mark the beginning of an Hif strain shift in Japan. PMID:27374690

  5. [A case of acute motor sensory axonal polyneuropathy after Haemophilus influenzae infection].

    Science.gov (United States)

    Oda, M; Udaka, F; Kubori, T; Oka, N; Kameyama, M

    2000-08-01

    A 47-year-old woman developed consciousness disturbance, and experienced hallucinations while traveling abroad, and then went into critical condition. She was placed in the critical care unit, and had flaccid tetraparesis requiring mechanical ventilation. Haemophilus influenzae was cultured from the sputum. The level of protein of the cerebrospinal fluid was elevated to 114 mg/dl, nerve conduction study showed findings of pure axonal damage, and the sural nerve biopsy revealed severe axonal degeneration. She improved gradually by plasma exchange. The diagnosis of acute motor sensory axonal polyneuropathy (AMSAN) based on autoimmune mechanism was made. We speculate that H. influenzae infection may have elicited AMSAN in this case. PMID:11218707

  6. Genetic Analysis of the Capsule Locus of Haemophilus influenzae Serotype f

    OpenAIRE

    Satola, Sarah W.; Patricia L Schirmer; Farley, Monica M.

    2003-01-01

    A 19-kb DNA region containing genes involved in the biosynthesis of the capsule of Haemophilus influenzae serotype f (Hif) has been cloned and characterized. The Hif cap locus organization is typical of group II capsule biosynthetic loci found in other H. influenzae serotype b bacteria and other gram-negative bacteria. However, the Hif cap locus was not associated with an IS1016 element. Three new open reading frames, Fcs1, Fcs2, and Fcs3, were identified in the Hif capsule-specific region II...

  7. Non-typeable Haemophilus influenzae purulent pericarditis in a child with cystic fibrosis.

    Science.gov (United States)

    Downes, Kevin J; Abulebda, Kamal; Siracusa, Christopher; Moore, Ryan; Staat, Mary A; Poynter, Sue E

    2016-07-01

    Early airway colonization and infection with Haemophilus influenzae in children with cystic fibrosis (CF) is common. Although the pathogenicity of non-typeable H. influenzae (NTHi) in patients with CF is controversial, this organism can cause both upper and lower respiratory tract infections. Extra-pulmonary disease, however, is rare. Purulent pericarditis is a suppurative complication of bacterial infection of the pericardial space that can arise as a result of direct extension from an adjacent infection. We describe a case of purulent pericarditis due to NTHi in a young child with CF that developed as a complication of inadequately treated bronchopneumonia.

  8. In Vitro Activity of Delafloxacin Tested against Isolates of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis.

    Science.gov (United States)

    Flamm, Robert K; Rhomberg, Paul R; Huband, Michael D; Farrell, David J

    2016-10-01

    Delafloxacin, an investigational anionic fluoroquinolone, is active against a broad range of Gram-positive and Gram-negative bacteria. In this study, 200 Streptococcus pneumoniae (plus 30 levofloxacin-resistant isolates), 200 Haemophilus influenzae, and 100 Moraxella catarrhalis isolates selected primarily from the United States (2014) were tested against delafloxacin and comparator agents. Delafloxacin was the most potent agent tested. MIC50 and MIC90 values against all S. pneumoniae isolates were 0.008 and 0.015 μg/ml. Delafloxacin susceptibility was not affected by β-lactamase status against H. influenzae and M. catarrhalis.

  9. Whole-genome random sequencing and assembly of Haemophilus influenzae Rd

    Energy Technology Data Exchange (ETDEWEB)

    Fleischmann, R.D.; Adams, M.D.; White, O. [Institute for Genomic Research, Gaithersburg, MD (United States)] [and others

    1995-07-28

    An approach for genome analysis based on sequencing and assembly of unselected pieces of DNA from the whole chromosome has been applied to obtain the complete nucleotide sequence (1,830,137 base pairs) of the genome from the bacterium Haemophilus influenzae Rd. This approach eliminates the need for initial mapping efforts and is therefore applicable to the vast array of microbial species for which genome maps are unavailable. The H. influenzae Rd genome sequence (Genome Sequence DataBase accession number L42023) represents the only complete genome sequence from a free-living organism. 46 refs., 4 figs., 4 tabs.

  10. Clonal relationship of recent invasive Haemophilus influenzae serotype f isolates from Denmark and the United States

    DEFF Research Database (Denmark)

    Bruun, B; Gahrn-Hansen, B; Westh, H;

    2004-01-01

    Surveillance performed after the introduction of general Haemophilus influenzae serotype b (Hib) vaccination in Denmark identified 13 cases of invasive bacteraemic H. influenzae serotype f (Hif) disease in adults over a period of 7 years. Bacteraemic respiratory tract infections accounted for 61...... % of cases, but meningitis, epiglottitis and osteoarthritis were also seen. Recent Danish isolates were compared to recent American isolates, historical Hif strains and non-Hif invasive strains. Results of conventional serotyping were confirmed by PCR detection of the serotype-f-specific cap and bexA gene...

  11. A glutathione-based system for defense against carbonyl stress in Haemophilus influenzae

    Directory of Open Access Journals (Sweden)

    Kidd Stephen P

    2012-07-01

    Full Text Available Abstract Background adhC from Haemophilus influenzae encodes a glutathione-dependent alcohol dehydrogenase that has previously been shown to be required for protection against killing by S-nitrosoglutathione (GSNO. This group of enzymes is known in other systems to be able to utilize substrates that form adducts with glutathione, such as aldehydes. Results Here, we show that expression of adhC is maximally induced under conditions of high oxygen tension as well as specifically with glucose as a carbon source. adhC could also be induced in response to formaldehyde but not GSNO. An adhC mutant was more susceptible than wild-type Haemophilus influenzae Rd KW20 to killing by various short chain aliphatic aldehydes, all of which can be generated endogenously during cell metabolism but are also produced by the host as part of the innate immune response. Conclusions These results indicate that AdhC plays a role in defense against endogenously generated reactive carbonyl electrophiles in Haemophilus influenzae and may also play a role in defense against the host innate immune system.

  12. Comparative Analyses of the Lipooligosaccharides from Nontypeable Haemophilus influenzae and Haemophilus haemolyticus Show Differences in Sialic Acid and Phosphorylcholine Modifications

    Science.gov (United States)

    Post, Deborah M. B.; Ketterer, Margaret R.; Coffin, Jeremy E.; Reinders, Lorri M.; Munson, Robert S.; Bair, Thomas; Murphy, Timothy F.; Foster, Eric D.; Gibson, Bradford W.

    2016-01-01

    Haemophilus haemolyticus and nontypeable Haemophilus influenzae (NTHi) are closely related upper airway commensal bacteria that are difficult to distinguish phenotypically. NTHi causes upper and lower airway tract infections in individuals with compromised airways, while H. haemolyticus rarely causes such infections. The lipooligosaccharide (LOS) is an outer membrane component of both species and plays a role in NTHi pathogenesis. In this study, comparative analyses of the LOS structures and corresponding biosynthesis genes were performed. Mass spectrometric and immunochemical analyses showed that NTHi LOS contained terminal sialic acid more frequently and to a higher extent than H. haemolyticus LOS did. Genomic analyses of 10 strains demonstrated that H. haemolyticus lacked the sialyltransferase genes lic3A and lic3B (9/10) and siaA (10/10), but all strains contained the sialic acid uptake genes siaP and siaT (10/10). However, isothermal titration calorimetry analyses of SiaP from two H. haemolyticus strains showed a 3.4- to 7.3-fold lower affinity for sialic acid compared to that of NTHi SiaP. Additionally, mass spectrometric and immunochemical analyses showed that the LOS from H. haemolyticus contained phosphorylcholine (ChoP) less frequently than the LOS from NTHi strains. These differences observed in the levels of sialic acid and ChoP incorporation in the LOS structures from H. haemolyticus and NTHi may explain some of the differences in their propensities to cause disease. PMID:26729761

  13. An epidemiological study of Haemophilus influenzae at a Brazilian day care center

    Directory of Open Access Journals (Sweden)

    M.E. Bonifácio da Silva

    2001-10-01

    Full Text Available Day care centers are a relatively new phenomenon in Brazil that bring together large numbers of young children susceptible to contagious diseases. Haemophilus influenzae (Hi is an important infection in the age range of those attending day care centers. In the present study, the carriage rate of Haemophilus influenzae was identified in 38 day care attendees age 6 to 37 months, and 23 staff members, at a day care center in Ribeirão Preto-São Paulo, in 1997. To identify the carriers, two nasopharyngeal swabs were collected; one in July and one in December. The rate of H. influenzae carriers among the children was 77%. Only 2 of 23 staff members (9% had Hi. Among the children, there were 58 isolates in the two sampling periods; 6 of the Hi were serotype b, 1 was serotype e, and 48 isolates were non-typeable. Two were identified as H. parainfluenzae. One adult had a non-typeable Hi and 1 had H. paraphrohaemolyticus. Three of the 6 children with type B had received a conjugate vaccine against H. influenzae type b, but they still carried this bacterium in the nasopharynx (50%. Forty ribotype patterns were found among the isolates, showing a high exchange rate of nontypeable H. influenzae carriers. The results indicate that, because of the high and changing biotype of Hi carriage, day care centers should be carefully monitored as potential point source of HI disease in the community.

  14. Cell vacuolation induced by Haemophilus influenzae supernatants in HEp-2 cells

    Directory of Open Access Journals (Sweden)

    Maria del Rosario Espinoza-Mellado

    2013-12-01

    Full Text Available Haemophilus influenzae belongs to respiratory tract microbiota. We observed vacuoles formation in previous studies with H. influenzae culture supernatants, so in this work we characterised that cytotoxic effect. We observed an abundant production of acidic cytoplasmic vacuoles due to the presence of a “vacuolating factor” in H. influenzae supernatants which was characterised as thermolabile. Greatest vacuolating activity was observed when utilizing the fraction > 50 kDa. The presence of a large number of vacuoles in HEp-2 cells was verified by transmission electron microscopy and some vacuoles were identified with a double membrane and/or being surrounded by ribosomes. These results suggest similar behaviour to that of vacuolating effects described by autotransporter proteins an undescribed cytotoxic effect induced by H. influenzae .

  15. Clinical characteristics of Haemophilus influenzae meningitis in Denmark in the post-vaccination era

    DEFF Research Database (Denmark)

    Pedersen, T.I.; Howitz, M.; Andersen, Christian Østergaard

    2010-01-01

    P>The introduction of Haemophilus influenzae type b (Hib) vaccine into the Danish childhood vaccination programme in 1993 may have influenced the epidemiology of H. influenzae meningitis (i.e. increasing frequency of other non-vaccine types; presentation in other age groups). Based on nationwide...... registration, clinical information and laboratory findings were collected from all 65 confirmed cases of H. influenzae meningitis during the period 1994-2005. Twenty-nine patients (45%) were 24 years old [median 62 years (range 25...... infected with Hib, two cases (13%) were identified as true vaccine failures. Six patients (9%) died; one premature infant infected with serotype f and five adults (age 83-96 years) with non-typeable H. influenzae. Hearing loss was reported in 16% of the surviving children and in 10% of the surviving adults...

  16. Serious systemic infection caused by non-encapsulated Haemophilus influenzae biotype III in an adult

    DEFF Research Database (Denmark)

    Lester, Anne; Pedersen, P B

    1991-01-01

    Haemophilus influenzae is the aetiological agent in less than 1% of septic arthritis cases in adults and most often serotype b is involved. We report here a case of severe systemic infection due to non-encapsulated H. influenzae biotype III in a 40-year-old man, previously healthy although alcohol...... abuser. Cholangitis and acute alcoholic hepatitis were diagnosed simultaneously. The organism was grown from blood and from synovial fluid of the left knee, but several other joints were also affected. The close relationship between H. influenzae biotype III and H. aegyptius is mentioned in view...... of recent reports of fatal childhood illness caused by a special clone of H. aegyptius and the importance of reporting both serotype and biotype in severe H. influenzae induced disease is emphasized....

  17. Detection of Cryptic Genospecies Misidentified as Haemophilus influenzae in Routine Clinical Samples by Assessment of Marker Genes fucK, hap, and sodC▿

    OpenAIRE

    Nørskov-Lauritsen, Niels

    2009-01-01

    Clinical isolates of Haemophilus influenzae were assessed for the presence of fucK, hap, and sodC by hybridization with gene-specific probes, and isolates diverging from the expected H. influenzae genotype were characterized by phenotype and 16S rRNA gene sequencing. Two of 480 isolates were finally classified as variant strains (“nonhemolytic Haemophilus haemolyticus”).

  18. Detection of cryptic genospecies misidentified as Haemophilus influenzae in routine clinical samples by assessment of marker genes fucK, hap, and sodC.

    Science.gov (United States)

    Nørskov-Lauritsen, Niels

    2009-08-01

    Clinical isolates of Haemophilus influenzae were assessed for the presence of fucK, hap, and sodC by hybridization with gene-specific probes, and isolates diverging from the expected H. influenzae genotype were characterized by phenotype and 16S rRNA gene sequencing. Two of 480 isolates were finally classified as variant strains ("nonhemolytic Haemophilus haemolyticus"). PMID:19535530

  19. Haemophilus haemolyticus interaction with host cells is different to nontypeable Haemophilus influenzae and prevents NTHi association with epithelial cells

    Directory of Open Access Journals (Sweden)

    Janessa Lea Pickering

    2016-05-01

    Full Text Available Nontypeable Haemophilus influenzae (NTHi is an opportunistic pathogen that resides in the upper respiratory tract and contributes to a significant burden of respiratory related diseases in children and adults. Haemophilus haemolyticus is a respiratory tract commensal that can be misidentified as NTHi due to high levels of genetic relatedness. There are reports of invasive disease from H. haemolyticus, which further blurs the species boundary with NTHi. To investigate differences in pathogenicity between these species, we optimized an in vitro epithelial cell model to compare the interaction of 10 H. haemolyticus strains with 4 NTHi and 4 H. influenzae-like haemophili. There was inter- and intra-strain variability but overall, H. haemolyticus had reduced capacity to attach to and invade nasopharyngeal and bronchoalveolar epithelial cell lines (D562 and A549 within 3h when compared with NTHi. H. haemolyticus was cytotoxic to both cell lines at 24h, whereas NTHi was not. Nasopharyngeal epithelium challenged with some H. haemolyticus strains released high levels of inflammatory mediators IL-6 and IL-8, whereas NTHi did not elicit an inflammatory response despite higher levels of cell association and invasion. Furthermore, peripheral blood mononuclear cells stimulated with H. haemolyticus or NTHi released similar and high levels of IL-6, IL-8, IL-10, IL-1β and TNFα when compared with unstimulated cells but only NTHi elicited an IFNγ response.Due to the relatedness of H. haemolyticus and NTHi, we hypothesized that H. haemolyticus may compete with NTHi for colonization of the respiratory tract. We observed that in vitro pre-treatment of epithelial cells with H. haemolyticus significantly reduced NTHi attachment, suggesting interference or competition between the two species is possible and warrants further investigation. In conclusion, H. haemolyticus interacts differently with host cells compared to NTHi, with different immunostimulatory and

  20. Oropharyngeal colonization by Haemophilus influenzae in healthy children from Taubaté (São Paulo), prior to the Haemophilus influenzae type b vaccination program in Brazil Colonização da orofaringe de crianças saudáveis de Taubaté (São Paulo) por Haemophilus influenzae, antes da introdução da vacina contra Haemophilus influenzae do tipo b no Brasil

    OpenAIRE

    Lucia Ferro Bricks; Caio Márcio Figueredo Mendes; Bianca Rezende Lucarevschi; Carmem Paz Oplustil; Zanella, Rosemeire C.; Adriana Bori; Ciro João Bertoli

    2004-01-01

    Haemophilus influenzae is one of the most important bacterial agents of otitis and sinusitis. H. influenzae type b (Hib) is one of the main causes of meningitis, pneumonia, and septicemia in nonvaccinated children under 6 years of age. The aims of this study were to determine the prevalence of H. influenzae and Hib oropharyngeal colonization prior to the onset of the Hib vaccination program in Brazil in previously healthy children and to assess the susceptibility profile of this microorganism...

  1. Identification of a DNA transformation gene required for com101A+ expression and supertransformer phenotype in Haemophilus influenzae.

    OpenAIRE

    Zulty, J J; Barcak, G J

    1995-01-01

    DNA sequencing, RNA mapping, and protein expression experiments revealed the presence of a gene, tfoX+, encoding a 24.9-kDa polypeptide, that is transcribed divergently from a common promoter region with the Haemophilus influenzae rec-1+ gene. H. influenzae strains mutant for tfoX failed to bind transforming DNA and were transformation deficient. Primer extension experiments utilizing in vivo total RNA from precompetent and competent H. influenzae cells demonstrated that transcription of tfoX...

  2. Pharmacokinetics and bacteriological efficacy of cefoperazone, ceftriaxone, and moxalactam in experimental Streptococcus pneumoniae and Haemophilus influenzae meningitis.

    OpenAIRE

    McCracken, G H; Nelson, J.D.; Grimm, L

    1982-01-01

    The pharmacokinetics and bacteriological efficacy of cefoperazone, cefuroxime, ceftriaxone, and moxalactam were evaluated in the experimental rabbit meningitis model of Haemophilus influenzae type b or Streptococcus pneumoniae infection. The cerebrospinal fluid penetration of these beta-lactam antibiotics was from 3 to 14% and was greater in Haemophilus-infected that in pneumococcus-infected animals. With the exception of moxalactam, the antibacterial activity in cerebrospinal fluid and chang...

  3. Clinical characteristics of Haemophilus influenzae meningitis in Denmark in the post-vaccination era

    DEFF Research Database (Denmark)

    Pedersen, T.I.; Howitz, Michael Frantz; Andersen, Christian Østergaard

    2010-01-01

    P>The introduction of Haemophilus influenzae type b (Hib) vaccine into the Danish childhood vaccination programme in 1993 may have influenced the epidemiology of H. influenzae meningitis (i.e. increasing frequency of other non-vaccine types; presentation in other age groups). Based on nationwide...... registration, clinical information and laboratory findings were collected from all 65 confirmed cases of H. influenzae meningitis during the period 1994-2005. Twenty-nine patients (45%) were 24 years old [median 62 years (range 25....... The presence of a lung focus was an independent prognostic factor for an unfavourable outcome (p 0.03). In conclusion, meningitis caused by Hib has been infrequent in Denmark after introduction of the Hib vaccine in the childhood vaccination programme, and no increase in meningitis cases due to non-b type H...

  4. Inflammatory response of Haemophilus influenzae biotype aegyptius causing Brazilian Purpuric Fever

    Science.gov (United States)

    Cury, Gisele Cristiane Gentile; Pereira, Rafaella Fabiana Carneiro; de Hollanda, Luciana Maria; Lancellotti, Marcelo

    2014-01-01

    The Brazilian Purpuric Fever (BPF) is a systemic disease with many clinical features of meningococcal sepsis and is usually preceded by purulent conjunctivitis. The illness is caused by Haemophilus influenza biogroup aegyptius, which was associated exclusively with conjunctivitis. In this work construction of the las gene, hypothetically responsible for this virulence, were fusioned with ermAM cassette in Neisseria meningitidis virulent strains and had its DNA transfer to non BPF H. influenzae strains. The effect of the las transfer was capable to increase the cytokines TNFα and IL10 expression in Hec-1B cells line infected with these transformed mutants (in eight log scale of folding change RNA expression). This is the first molecular study involving the las transfer to search an elucidation of the pathogenic factors by horizontal intergeneric transfer from meningococci to H. influenzae. PMID:25763053

  5. Inflammatory response of Haemophilus influenzae biotype aegyptius causing Brazilian Purpuric Fever

    Directory of Open Access Journals (Sweden)

    Gisele Cristiane Gentile Cury

    2014-12-01

    Full Text Available The Brazilian Purpuric Fever (BPF is a systemic disease with many clinical features of meningococcal sepsis and is usually preceded by purulent conjunctivitis. The illness is caused by Haemophilus influenza biogroup aegyptius, which was associated exclusively with conjunctivitis. In this work construction of the las gene, hypothetically responsible for this virulence, were fusioned with ermAM cassette in Neisseria meningitidis virulent strains and had its DNA transfer to non BPF H. influenzae strains. The effect of the las transfer was capable to increase the cytokines TNFα and IL10 expression in Hec-1B cells line infected with these transformed mutants (in eight log scale of folding change RNA expression. This is the first molecular study involving the las transfer to search an elucidation of the pathogenic factors by horizontal intergeneric transfer from meningococci to H. influenzae.

  6. Duplex Quantitative PCR Assay for Detection of Haemophilus influenzae That Distinguishes Fucose- and Protein D-Negative Strains.

    Science.gov (United States)

    de Gier, Camilla; Pickering, Janessa L; Richmond, Peter C; Thornton, Ruth B; Kirkham, Lea-Ann S

    2016-09-01

    We have developed a specific Haemophilus influenzae quantitative PCR (qPCR) that also identifies fucose-negative and protein D-negative strains. Analysis of 100 H. influenzae isolates, 28 Haemophilus haemolyticus isolates, and 14 other bacterial species revealed 100% sensitivity (95% confidence interval [CI], 96% to 100%) and 100% specificity (95% CI, 92% to 100%) for this assay. The evaluation of 80 clinical specimens demonstrated a strong correlation between semiquantitative culture and the qPCR (P < 0.001). PMID:27335148

  7. Identification of a surface-exposed immunodominant epitope on outer membrane protein P1 of Haemophilus influenzae type b.

    OpenAIRE

    Proulx, C; Munson, R S; Grass, S; Hamel, J; Martin, D.; Brodeur, B R

    1991-01-01

    Eight murine monoclonal antibodies (MAbs) directed against outer membrane protein P1 of Haemophilus influenzae type b were generated and characterized. Seven of the eight MAbs reacted with recombinant P1 and purified P1 protein from H. influenzae type b strains MinnA and 1613; MAb P1.8 was specific for the latter strain. A panel of 32 nontypeable and 140 encapsulated Haemophilus strains recovered worldwide representing the major clonal families of serotypes a, b, and d was used to evaluate th...

  8. Vacinação contra Haemophilus influenzae tipo b: proteção a longo prazo Haemophilus influenzae type b vaccination: long term protection

    Directory of Open Access Journals (Sweden)

    Cristiana M. Nascimento-Carvalho

    2006-07-01

    Full Text Available OBJETIVO: Identificar as evidências sobre o impacto da vacina conjugada para Haemophilus influenzae tipo b (Hib na epidemiologia da doença invasiva por Hib. FONTE DOS DADOS: Pesquisa nas bases de dados do MEDLINE, LILACS, publicações técnicas de organizações internacionais, diretrizes nacionais e internacionais, nos últimos 15 anos (1991-2005, utilizando os seguintes unitermos: Haemophilus influenzae type b, immunization, impact, effectiveness. Foram incluídas as publicações que apresentaram informação para atender o objetivo deste artigo. Artigos publicados em período anterior ao da pesquisa e citados em referências dos artigos incluídos foram analisados quanto à apresentação de informação de interesse. SÍNTESE DOS DADOS: A introdução da vacina conjugada para Hib produziu grande declínio na incidência de casos de doença invasiva por Hib nos diversos países em que seu uso foi incorporado à rotina de vacinação das crianças. No entanto, o ressurgimento de casos com doença invasiva por Hib tem mobilizado vários investigadores na busca das possíveis explicações para esses eventos, bem como a identificação das medidas a serem implementadas para evitar o reaparecimento da doença. CONCLUSÕES: O uso da vacina conjugada para Hib em escala populacional tem sido extremamente efetivo. No entanto, mudanças no esquema vacinal poderão ser necessárias para a manutenção do controle da doença invasiva por Hib, frente ao atual cenário epidemiológico das infecções pelo Hib.OBJECTIVE: To identify evidence of the impact of Haemophilus influenzae type b (Hib conjugate vaccine on the epidemiology of invasive Hib disease. SOURCES OF DATA: This review was based on a search of MEDLINE, LILACS, technical reports, national and international guidelines (publications from 1991 to 2005. The keywords Haemophilus influenzae type b, immunization, impact and effectiveness, alone or in combination, were used to retrieve the

  9. Meningite por Haemophilus influenzae tipo b em cidades do estado do Paraná, Brasil Haemophilus influenzae type b meningitis in the state of Paraná, Brazil

    Directory of Open Access Journals (Sweden)

    Nádia S. Takemura

    2001-10-01

    Full Text Available OBJETIVO: no segundo semestre de 1996, os municípios de Londrina e Curitiba (Paraná iniciaram a vacinação contra Haemophilus influenzae b (Hib, aproximadamente trinta meses antes de sua introdução no Programa Nacional de Imunização. O presente trabalho objetivou avaliar a incidência da meningite por Hib, entre crianças, em Londrina, Curitiba e nos demais municípios do estado do Paraná, antes e após a introdução da vacina nessas duas cidades. MÉTODOS: foi realizado um estudo observacional retrospectivo de todos os casos de meningite por Hib, entre menores de 5 anos, diagnosticados pelo sistema de vigilância epidemiológica de Londrina e pela Secretaria de Estado da Saúde do Paraná, de 1992 a 1999. Taxas de incidência da meningite por Hib foram calculadas por 100.000 menores de cinco anos. RESULTADOS: comparando com o período anterior à vacinação, houve redução importante do coeficiente de incidência da meningite por Hib em Londrina, passando de 23,91, em 1996, para 2,79 por 100.000 menores de cinco anos, em 1999. Redução semelhante foi observada em Curitiba, enquanto nos demais municípios do Paraná, que não dispunham da vacina até meados de 1999, o coeficiente se manteve praticamente inalterado. CONCLUSÃO: a vacinação contra Hib foi efetiva na redução da incidência da meningite entre menores de cinco anos em Londrina e Curitiba. Para a manutenção dessa baixa incidência devem ser garantidas adequada cobertura vacinal e boa qualidade do serviço de vigilância epidemiológica.OBJECTIVE: during the second half of 1996, the municipalities of Londrina and Curitiba (State of Paraná, Brazil included Haemophilus influenzae type b (Hib vaccine into their routine vaccination regimen, approximately 30 months before its introduction into the National Immunization Program. The present study aimed at verifying the incidence of meningitis caused by Hib among children in Londrina, Curitiba, and in the remaining

  10. Genotyping of Haemophilus influenzae type b strains and their incidence in the clinical samples isolated from Iranian patients

    Science.gov (United States)

    Bagherzadeh Khodashahri, Somayeh; Siadat, Seyed Davar; Rahbar, Mohammad; Abdollahpour-Alitappeh, Meghdad; Vaziri, Farzam; Rahnamaye-Farzami, Mrjan; Mohammadzadeh, Mona; Davari, Mehdi; Fateh, Abolfazl; Masoumi, Morteza

    2015-01-01

    Background and Objective: Haemophilus influenzae type b (Hib) is divided into two distinct genotypes, type I and type II, based on the structure of capsular polysaccharides. The capsulation locus of Haemophilus influenzae type b consists of three functionally distinct regions, designated regions 1 to 3. Region III contains hcsA and hcsB genes; however, notable sequence variation in this region can be used to recognize different Hib genotypes. The purpose of this study was to investigate the prevalence and genotype of the Hib strains isolated from patients with invasive disease in Iran. Materials and Methods: In the present study, 8 pairs of primers were used for identification and serotyping of encapsulated Haemophilus influenzae strains, as well as confirmation of species identification. Additionally, in order to identify the capsular genotypes of Haemophilus influenzae type b (type I and II), two additional primer pairs were used to amplify the hcsA gene. Results: Out of 50 isolates of H. influenzae, four were found to be type b. Interestingly, among these 4 Hib isolates, 2 strains belonged to the type-II category. Conclusion: Our study shows that the prevalence of both Hib types I and II seems to be high in Iran. PMID:26668700

  11. Evolution of the paralogous hap and iga genes in Haemophilus influenzae: evidence for a conserved hap pseudogene associated with microcolony formation in the recently diverged Haemophilus aegyptius and H. influenzae biogroup aegyptius

    DEFF Research Database (Denmark)

    Kilian, Mogens; Poulsen, Knud; Lomholt, Hans Bredsted

    2002-01-01

    Certain non-capsulate strains belonging to the Haemophilus influenzae/Haemophilus aegyptius complex show unusually high pathogenicity, but the evolutionary origin of these virulent phenotypes, termed H. influenzae biogroup aegyptius, is as yet unknown. The aim of the present study was to elucidate...... the mechanisms of evolution of two paralogous genes, hap and iga, which encode the adhesion and penetration Hap protein and the IgA1 protease respectively. Partial sequencing of hap and iga genes in a comprehensive collection of strains belonging to the H. influenzae/H. aegyptius complex revealed considerable...... genetic polymorphism and pronounced mosaic-like patterns in both genes, but no evidence of intrastrain recombination between the two genes. A conserved hap pseudogene was present in all strains of H. aegyptius and H. influenzae biogroup aegyptius, each of which constituted distinct subpopulations...

  12. Capsulation and Gene Copy Number at the cap Locus of Haemophilus influenzae Type b

    OpenAIRE

    1988-01-01

    Although more than 98% of natural isolates of Haemophilus influenzae type b carry a duplication of 17 kilobases (kb) of DNA at the chromosomal capsulation locus, only one copy is required for capsulation. In one laboratory-derived and two clinical type b strains, the capsulation locus had a single copy of this 17-kb segment, together with 1.3 kb of DNA identified as lying between the repeats of the duplicated locus. This 1.3 kb appears to be crucial for capsule production, since strains lacki...

  13. Phylogenetic relationship of non-typeable Haemophilus influenzae isolated in Malaysia.

    Science.gov (United States)

    Mohd-Zain, Zaini; Kamsani, Nurul H; Ahmad, Norazah; Clarke, Stuart C

    2015-12-01

    The epidemiology of non-typeable Haemophilus influenzae (NTHi) remains poorly understood. We therefore sought to determine the genetic relationship of 25 NTHi isolated from various states in Malaysia using multilocus sequence typing (MLST). The majority of isolates were obtained from sputum. There were 24 novel sequence types (STs). Eight isolates were single-locus variants, the remainder being singletons. Clustering was not based on clinical site of isolation or geographical origin. Despite the limited number of isolates examined in this study, we demonstrate that NTHi isolates in Malaysia are diverse and warrant further investigation.

  14. A biphasic epigenetic switch controls immunoevasion, virulence and niche adaptation in non-typeable Haemophilus influenzae

    OpenAIRE

    Atack, John M.; Srikhanta, Yogitha N.; Fox, Kate L.; Jurcisek, Joseph A.; Brockman, Kenneth L.; Tyson A Clark; Boitano, Matthew; Power, Peter M.; Jen, Freda E.-C.; McEwan, Alastair G; Grimmond, Sean M.; Smith, Arnold L.; Barenkamp, Stephen J.; Korlach, Jonas; Bakaletz, Lauren O.

    2015-01-01

    Non-typeable Haemophilus influenzae contains an N6-adenine DNA-methyltransferase (ModA) that is subject to phase-variable expression (random ON/OFF switching). Five modA alleles, modA2, modA4, modA5, modA9 and modA10, account for over two-thirds of clinical otitis media isolates surveyed. Here, we use single molecule, real-time (SMRT) methylome analysis to identify the DNA-recognition motifs for all five of these modA alleles. Phase variation of these alleles regulates multiple proteins inclu...

  15. [Isolation of Haemophilus influenzae serotypes from deep sites in sick children].

    Science.gov (United States)

    Gatti, B M; Ramirez Gronda, G A; Etchevarría, M; Vescina, C M; Varea, A M; González Ayala, S E

    2004-01-01

    Haemophilus influenzae (Hi) is the causative agent of several human diseases such as sepsis, meningitis, celulitis, and osteoarthritis. We investigated the isolation of Hi serotypes from sterile sites in sick children. One hundred and seventy nine strains from 146 patients were studied, period 1996-2002, at the Microbiology Laboratory, Hospital de Niños Superiora Sor María Ludovica, Argentina. The serotype distribution was:1 a, 112 b,1 c,1 d, 4 e, 3 f y 24 no typable. Since the beginning of universal Hi b vaccination in 1998, we have observed the fast decrease of serotype b and a relative increase of other serotypes.

  16. Similarity in properties and mapping of three rec mutants of Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Kooistra, J.; Setlow, J.K.

    1976-07-01

    Three Rec/sup -/ mutants of Haemophilus influenzae have been studied with respect to their transformability, ultraviolet and mitomycin C sensitivities, spontaneous and ultraviolet-induced deoxyribonucleic acid breakdown, inducibility of lysogens, and the linkage of the three mutations to a streptomycin resistance marker. The data indicate that the three mutations cause the same phenotypic changes, and that they are all on the same gene. Transformability of the mutants is different when two different media are used for competence development, although transformability with the two competence methods is not different in a Rec/sup -/ strain that is mutant at another gene.

  17. Repair of ultraviolet-irradiated transforming DNA in A recA mutant of Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Stuy, J.H.; Walter, R.B. (Florida State Univ., Tallahassee (USA). Dept. of Biological Science)

    1983-04-01

    Ultraviolet-irradiated transforming DNA was assayed on a wild-type strain of Haemophilus influenzae strain Rd, on an excision repair-deficient (uvr-2) mutant, on a recombination repair-deficient (recA4) mutant, and on a strain carrying both mutations. The donor DNA had a point mutation genetic marker (strAl) and a long nonhomologous plasmid-derived DNA segment inserted in the HPl prophage. The shape of the inactivation curves suggested that only recombination was responsible for the inverse square root kinetics observed with excision repair-proficient recipients.

  18. Special type of UV-stimulated recombination in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Waldstein, E; Setlow, J K; Santasier, L

    Following uv irradiation, the DNA of wild type H. influenzae undergoes extensive exchange between strands synthesized before and after irradiation. There are apparently many more exchanges than uv-induced pyrimidine dimers, unlike the situation in Esherichia coli. Such uv-induced exchange does not take place in the highly recombination-defective rec1 strain of H. influenzae, but is extensive in the even more recombination-defective rec2 strain. These events are correlated with the filling of the single-strand gaps made in DNA synthesized after uv; in rec2 the gaps are filled normally, but there is little or no gapfilling in rec1. Normal DNA replication is not required for gapfilling, since it takes place in a temperature-sensitive DNA synthesis mutant at a temperature restrictive for DNA synthesis. Thus it is probable that in H. influenzae exchange is more important for gapfilling than is de novo synthesis. In order to find out more about uv-induced exchange, we have examined the question of whether dimers originally in DNA synthesized before irradiation could be shifted into the DNA made after irradiation by way of such a mechanism.

  19. Antibiotic resistance of streptococcus pneumoniae and haemophilus influenzae isolated from respiratory tract specimens

    Directory of Open Access Journals (Sweden)

    Hikmet Eda Aliskan

    2016-06-01

    Full Text Available Purpose: Streptococcus pneumoniae and Haemophilus influenzae are two of the major pathogens in respiratory infections, treatment is usually started empirically. The aim of this study was to detect in vitro resistance rates of S. pneumoniae and H. influenzae strains isolated from different lower respiratory clinical samples to the antibotics which are used for therapy of infections due to these pathogens. Material and Methods: Seventy seven S.pneumoniae and 117 H.influenzae strains, isolated from patients were included in the study. S.pneumoniae isolates which gave an inhibition zone diameter of >20 mm for oxacillin were considered susceptible for penicilin. For the isolates which had an oxacillin zone diameter of 2 mg/l and 31.1 % were intermediately resistant to parenteral penicillin. Resistance rates to antibiotics were as follows: erythromycin 40 %, trimethoprim/sulphametoxazole (TMP/SMX 54.5 % and ofloxacin 6.4%. beta-lactamases were detected in 15.6% of the H.influenzae isolates by nitrocefin positivity. Conclusion: H.influenzae strains (8.6% were identified as beta-lactamase negative ampicillin resistant (BLNAR strains. Resistance rates for other antibiotics were as follows: ampicillin 28.6%, cefaclor 36.5% , cefuroxime 30.1%, clarithromycin 9.6%, cloramphenicol 7% and TMP-SMX 43.9%. [Cukurova Med J 2016; 41(2.000: 201-207

  20. DNA repair in Haemophilus influenzae: isolation and characterization of an ultraviolet sensitive mutator mutant

    Energy Technology Data Exchange (ETDEWEB)

    Walter, R.B.

    1985-01-01

    DNA repair in Haemophilus influenzae appears to be quite different from that seen in Escherichia coli in that H. influenzae shows neither SOS nor adaptation phenomena. Repair of DNA lesions in H. influenzae has been seen to occur via recombinational, excision, and mismatch repair pathways acting independently of one another. The author has isolated an ultraviolet (UV)-sensitive mutator mutant (mutB1) of H. influenzae Rd which shows deficiencies in both recombinational and mismatch repair pathways. This mutant is sensitive to a variety of DNA damaging agents as well as being hypermutable by alkylating agents and base analogues. MutB1 cells do not show post-UV DNA breakdown but do begin excision after UV irradiation. Genetic transformation with UV-irradiated DNA on mut B1 recipients shows that high (HE) and low (LE) efficiency markers are transformed at a ratio of 1.0 as in the mismatch repair deficient hex 1 mutant; however, kinetics of UV-inactivation experiments indicate that HE markers are sensitized and act as LE markers do on wild type recipients. Thus, the mutB gene product appears to play a role in both DNA repair and genetic transformation. A model is outlined which presents a role for a DNA helicase in both DNA repair and genetic transformation of H. influenzae.

  1. Haemophilus influenzae P4 Interacts With Extracellular Matrix Proteins Promoting Adhesion and Serum Resistance.

    Science.gov (United States)

    Su, Yu-Ching; Mukherjee, Oindrilla; Singh, Birendra; Hallgren, Oskar; Westergren-Thorsson, Gunilla; Hood, Derek; Riesbeck, Kristian

    2016-01-15

    Interaction with the extracellular matrix (ECM) is one of the successful colonization strategies employed by nontypeable Haemophilus influenzae (NTHi). Here we identified Haemophilus lipoprotein e (P4) as a receptor for ECM proteins. Purified recombinant P4 displayed a high binding affinity for laminin (Kd = 9.26 nM) and fibronectin (Kd = 10.19 nM), but slightly less to vitronectin (Kd = 16.51 nM). A P4-deficient NTHi mutant showed a significantly decreased binding to these ECM components. Vitronectin acquisition conferred serum resistance to both P4-expressing NTHi and Escherichia coli transformants. P4-mediated bacterial adherence to pharynx, type II alveolar, and bronchial epithelial cells was mainly attributed to fibronectin. Importantly, a significantly reduced bacterial infection was observed in the middle ear of the Junbo mouse model when NTHi was devoid of P4. In conclusion, our data provide new insight into the role of P4 as an important factor for Haemophilus colonization and subsequent respiratory tract infection.

  2. Structural diversity of the lipid A and core oligosaccharide moieties of the lipopolysaccharides from nontypeable and serotype f Haemophilus influenzae

    OpenAIRE

    Yildirim, Håkan

    2005-01-01

    This thesis describes structural studies of the oligosaccharide and lipid A moieties of lipopolysaccharides (LPSs) isolated from disease-causing Haemophilus influenzae strains. The nontypeable strains were clinical isolates from the middle ear of children suffering from otitis media and the serotype f strains had been collected from three adults with respiratory tract infections. The LPS molecules are situated on the cell wall of H. influenzae strains and they play a ver...

  3. Naturally acquired immunity to Haemophilus influenzae type B in healthy Cuban children

    Directory of Open Access Journals (Sweden)

    Gilda Toraño Peraza

    2004-11-01

    Full Text Available We have evaluated the prevalence of antibody to immunogenicity of Haemophilus influenzae type b (Hib in a group of 4 to 5 years old healthy children, who were too old to be included in the first vaccinated cohort when Hib vaccination begun in Cuba in 1999. Serum capsular polysaccharide specific IgG antibody concentrations were measured in 974 healthy children, between February and May 2002. The prevalence of Hib nasopharyngeal carriage was also estimated. The majority of children (99.7% had more than 1 µg/ml of antibody. The preliminary report of the nasopharyngeal cultures was positive for H. influenzae in 16 children, but in only one was confirmed as Hib after serotyping (0.1% Hib nasopharyngeal carrier. These results provide evidence that in Cuba the natural active immunity to Hib can be acquired at an early age.

  4. Experimental otitis media in gerbils and chinchillas with Streptococcus pneumoniae, Haemophilus influenzae, and other aerobic and anaerobic bacteria.

    OpenAIRE

    Fulghum, R S; Brinn, J E; Smith, A M; Daniel, H J; Loesche, P J

    1982-01-01

    To ascertain the usefulness of Mongolian gerbils as an inbred model for otitis media, 52 Mongolian gerbils (Meriones unguiculatus, strain MONT/Tum) were compared with 26 chinchillas (Chinchilla laniger) for susceptibility to Streptococcus pneumoniae type 3. Haemophilus influenzae type b, and a polymicrobic culture including anaerobes (Streptococcus intermedius, Propionibacterium acnes, Staphylococcus epidermidis, and Corynebacterium sp.). Organisms were inoculated percutaneously into the supe...

  5. Towards a sustainable, quality and affordable Haemophilus influenzae type b vaccine for every child in the world

    NARCIS (Netherlands)

    Hamidi, A.

    2016-01-01

    Haemophilus influenzae type b (Hib) conjugate vaccine is a safe and effective vaccine that can prevent meningitis and pneumonia caused by Hib disease. Hib vaccine is recommended for all children under 5 years. Despite the availability of safe and effective Hib vaccines since early 1987, Gambia was t

  6. [Haemophilus influenzae type b in Italy--after thirty years of vaccination may we lower our guard?].

    Science.gov (United States)

    Terracciano, Elisa; Zaratti, Laura; Franco, Elisabetta

    2015-01-01

    Haemophilus influenzae b (Hib) is responsible for meningitis, systemic infections and acute respiratory illness, especially in children. The use of the conjugate vaccines against Hib reduced the incidence of the disease worldwide. In Italy, after the decrease resulted from vaccination, the disease may reappear due to the reduction in vaccination coverage, the presence of infections in adults and vaccine failures. PMID:26722830

  7. Outbreak of amoxicillin-resistant Haemophilus influenzae type b: variable number of tandem repeats as novel molecular markers

    NARCIS (Netherlands)

    A.F. van Belkum (Alex); W.J. Melchers; C. IJsseldijk; L. Nohlmans; J.F. Meis; H.A. Verbrugh (Henri)

    1997-01-01

    textabstractAn outbreak caused by amoxicillin-resistant Haemophilus influenzae type b was noted among patients suffering from chronic obstructive pulmonary disease. Since infections were clustered in time and place, an ongoing outbreak was suspected. The spread of the s

  8. Identification of a group of Haemophilus influenzae penicillin-binding proteins that may have complementary physiological roles

    Energy Technology Data Exchange (ETDEWEB)

    Malouin, F.; Parr, T.R. Jr.; Bryan, L.E. (Eli Lilly Company, Indianapolis, IN (USA))

    1990-02-01

    (35S)penicillin bound to different Haemophilus influenzae proteins in assays performed at 20, 37, or 42{degrees}C. Penicillin-binding proteins 3a, 3b, 4, and 4' formed a group characterized by their affinity for moxalactam, cefotaxime, and piperacillin. Penicillin-binding protein 4' showed specific properties that may reflect its complementary role in septation.

  9. Fulminant bilateral Haemophilus influenzae keratitis in a patient with hypovitaminosis A treated with contaminated autologous serum

    Directory of Open Access Journals (Sweden)

    Empar Sanz-Marco

    2011-01-01

    Full Text Available Empar Sanz-Marco1, Manuel Diaz-Llopis1–3, Maria J Lopez-Prats1, Salvador Garcia-Delpech1,4, Patricia Udaondo11Ophthalmology, La Fe University Hospital of Valencia, Valencia, Spain; 2Faculty of Medicine, University of Valencia, Valencia, Spain; 3Biomedical Research Center for Rare Diseases Network (CIBERER, Valencia, Spain; 4Faculty of Medicine, Catholic University of Valencia, Valencia, SpainAbstract: We report on a patient with hypovitaminosis A treated with autologous serum (AS who had bilateral infectious ulcers positive for Haemophilus influenzae. The patient suffered a perforation of his right eye and total healing of his left eye with a residual leukoma. In cases of severe malnutrition, serum levels of vitamins and bacteriostatic and growth factors are reduced, so AS would not only be ineffective but also increase the risk of secondary corneal infection. The prophylactic use of a topical antibiotic would be useful in treatment with AS, especially in patients who do not use and adequately store the eye drops, as in our patient.Keywords: Haemophilus influenzae, hypovitaminosis A, bilateral infectious ulcers

  10. Structure of the N-terminal region of Haemophilus Influenzae HI0017: Implications for function

    Energy Technology Data Exchange (ETDEWEB)

    Yu Liping; Mack, Jamey; Hajduk, Phil; Fesik, Stephen W. [Abbott Laboratories, Pharmaceutical Discovery Division, D46Y, AP10/LL (United States)

    2001-06-15

    Haemophilus influenzae is a gram-negative pathogen that causes infections ranging from asymptomatic colonization of the human upper respiratory tract to serious invasive diseases such as meningitis. Although the genome of Haemophilus influenzae has been completely sequenced, the structure and function of many of these proteins are unknown. HI0017 is one of these uncharacterized proteins. Here we describe the three-dimensional solution structure of the N-terminal portion of HI0017 as determined by NMR spectroscopy. The structure consists of a five-stranded antiparallel {beta}-sheet and two short {alpha}-helices. It is similar to the C-terminal domain of Diphtheria toxin repressor (DtxR). The C-terminal portion of HI0017 has an amino acid sequence that closely resembles pyruvate formate-lyase - an enzyme that converts pyruvate and CoA into acetyl-CoA and formate by a radical mechanism. Based on structural and sequence comparisons, we propose that the C-terminus of HI0017 functions as an enzyme with a glycyl radical mechanism, while the N-terminus participates in protein/protein interactions involving an activase (iron-sulfur protein) and/or the substrate.

  11. Why we need a vaccine for non-typeable Haemophilus influenzae.

    Science.gov (United States)

    Cerquetti, Marina; Giufrè, Maria

    2016-09-01

    Nontypeable Haemophilus influenzae (NTHi) is increasingly recognized as emerging pathogen. The routine immunization of infants with conjugated vaccines against H. influenzae type b (Hib) has greatly reduced the incidence of invasive Hib disease; however a marked change in the predominant invasive serotype from Hib to NTHi has occurred. Localized infections where the role of H. influenzae is important, such as otitis media in children and acute exacerbations in chronic obstructive pulmonary disease (COPD) in adults, are almost exclusively associated with NTHi isolates. The implementation of pneumococcal conjugate vaccines has resulted in changes in frequency of nasopharynx colonizing pathogens with an increase of NTHi, although this data is yet under debate. An effective vaccine against NTHi is not currently available. The major challenge in developing a successful vaccine is the intrinsic heterogeneity of NTHi. H. influenzae protein D is used as carrier protein in the licensed 10-valent pneumococcal conjugate vaccine (Synflorix, GlaxoSmithKline), but no robust evidences for protective efficacy against NTHi otitis have been until now obtained. Several other vaccine candidates are under investigations and we hope that significant advancements in vaccine development will be achieved in the next future. Genome-based vaccine strategy might provide an additional useful tool for discovering further vaccine antigens.

  12. Molecular surveillance of true nontypeable Haemophilus influenzae: an evaluation of PCR screening assays.

    Directory of Open Access Journals (Sweden)

    Michael J Binks

    Full Text Available BACKGROUND: Unambiguous identification of nontypeable Haemophilus influenzae (NTHi is not possible by conventional microbiology. Molecular characterisation of phenotypically defined NTHi isolates suggests that up to 40% are Haemophilus haemolyticus (Hh; however, the genetic similarity of NTHi and Hh limits the power of simple molecular techniques such as PCR for species discrimination. METHODOLOGY/PRINCIPAL FINDINGS: Here we assess the ability of previously published and novel PCR-based assays to identify true NTHi. Sixty phenotypic NTHi isolates, classified by a dual 16S rRNA gene PCR algorithm as NTHi (n = 22, Hh (n = 27 or equivocal (n = 11, were further characterised by sequencing of the 16S rRNA and recA genes then interrogated by PCR-based assays targeting the omp P2, omp P6, lgtC, hpd, 16S rRNA, fucK and iga genes. The sequencing data and PCR results were used to define NTHi for this study. Two hpd real time PCR assays (hpd#1 and hpd#3 and the conventional iga PCR assay were equally efficient at differentiating study-defined NTHi from Hh, each with a receiver operator characteristic curve area of 0.90 [0.83; 0.98]. The hpd#1 and hpd#3 assays were completely specific against a panel of common respiratory bacteria, unlike the iga PCR, and the hpd#3 assay was able to detect below 10 copies per reaction. CONCLUSIONS/SIGNIFICANCE: Our data suggest an evolutionary continuum between NTHi and Hh and therefore no single gene target could completely differentiate NTHi from Hh. The hpd#3 real time PCR assay proved to be the superior method for discrimination of NTHi from closely related Haemophilus species with the added potential for quantification of H. influenzae directly from specimens. We suggest the hpd#3 assay would be suitable for routine NTHi surveillance and to assess the impact of antibiotics and vaccines, on H. influenzae carriage rates, carriage density, and disease.

  13. Invasive Haemophilus influenzae Serotype f Case Reports in Mazovia Province, Poland.

    Science.gov (United States)

    Golebiewska, Anna; Kuch, Alicja; Gawrońska, Agnieszka; Albrecht, Piotr; Skoczyńska, Anna; Radzikowski, Andrzej; Kutylowska, Ewa; Feleszko, Wojciech

    2016-02-01

    After successful introduction of anti-Haemophilus influenzae (Hi) serotype b vaccination program in Poland, invasive non-b or nontypeable H. influenzae infections have been reported more frequently alike in other countries all over the world. In this paper, we report 2 cases of H. influenzae serotype f (Hif) meningitis with severe clinical presentations which are rarely seen in previously healthy children.The first case is a 6-year-old girl who was admitted to pediatric ward with signs of meningitis. Laboratory tests confirmed bacteremic meningitis caused by Hif. The girl responded very well to administered treatment and recovered without any further complications. No underlying comorbidities were found. The second patient was a 4-year-old boy who, in course of Hif bacteremic meningitis, developed rapid septicemia and, despite aggressive treatment, died within a few hours of hospitalization. The child's past history was unremarkable.By presenting these cases, we would like to remind clinicians that invasive non-b Hi infections can become fatal not only in the group of the youngest children or children with coexisting comorbidities, as most commonly reported in the worldwide literature. At the same time, we want to emphasize the legitimacy of constant monitoring Hi epidemiology in order to take accurate actions if necessary. PMID:26844500

  14. Invasive Haemophilus influenzae Serotype f Case Reports in Mazovia Province, Poland.

    Science.gov (United States)

    Golebiewska, Anna; Kuch, Alicja; Gawrońska, Agnieszka; Albrecht, Piotr; Skoczyńska, Anna; Radzikowski, Andrzej; Kutylowska, Ewa; Feleszko, Wojciech

    2016-02-01

    After successful introduction of anti-Haemophilus influenzae (Hi) serotype b vaccination program in Poland, invasive non-b or nontypeable H. influenzae infections have been reported more frequently alike in other countries all over the world. In this paper, we report 2 cases of H. influenzae serotype f (Hif) meningitis with severe clinical presentations which are rarely seen in previously healthy children.The first case is a 6-year-old girl who was admitted to pediatric ward with signs of meningitis. Laboratory tests confirmed bacteremic meningitis caused by Hif. The girl responded very well to administered treatment and recovered without any further complications. No underlying comorbidities were found. The second patient was a 4-year-old boy who, in course of Hif bacteremic meningitis, developed rapid septicemia and, despite aggressive treatment, died within a few hours of hospitalization. The child's past history was unremarkable.By presenting these cases, we would like to remind clinicians that invasive non-b Hi infections can become fatal not only in the group of the youngest children or children with coexisting comorbidities, as most commonly reported in the worldwide literature. At the same time, we want to emphasize the legitimacy of constant monitoring Hi epidemiology in order to take accurate actions if necessary.

  15. Developing a vaccine to prevent otitis media caused by nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Khan, M Nadeem; Ren, Dabin; Kaur, Ravinder; Basha, Saleem; Zagursky, Robert; Pichichero, Michael E

    2016-07-01

    Nontypeable Haemophilus influenzae (NTHi) is a predominant organism of the upper respiratory nasopharyngeal microbiota. Its disease spectrum includes otitis media, sinusitis, non-bacteremic pneumonia and invasive infections. Protein-based vaccines to prevent NTHi infections are needed to alleviate these infections in children and vulnerable populations such as the elderly and those with chronic obstructive pulmonary disease (COPD). One NTHi protein is included in a pneumococcal conjugate vaccine and has been shown to provide efficacy. Our lab has been interested in understanding the immunogenicity of NTHi vaccine candidates P6, protein D and OMP26 for preventing acute otitis media in young children. We expect that continued investigation and progress in the development of an efficacious protein based vaccine against NTHi infections is achievable in the near future. PMID:26894630

  16. Vaccination for the control of childhood bacterial pneumonia - Haemophilus influenzae type b and pneumococcal vaccines

    Directory of Open Access Journals (Sweden)

    Diana C Otczyk

    2013-01-01

    Full Text Available Pneumonia in childhood is endemic in large parts of the world and in particular, in developing countries, as well as in many indigenous communities within developed nations. Haemophilus influenzae type b and Streptococcus pneumoniae conjugate vaccines are currently available against the leading bacterial causes of pneumonia.  The use of the vaccines in both industrialised and developing countries have shown a dramatic reduction in the burden of pneumonia and invasive disease in children.  However, the greatest threat facing pneumococcal conjugate vaccine effectiveness is serotype replacement.  The current vaccines provide serotype-specific, antibody–mediated protection against only a few of the 90+ capsule serotypes.  Therefore, there has been a focus in recent years to rapidly advance technologies that will result in broader disease coverage and more affordable vaccines that can be used in developing countries.  The next generation of pneumococcal vaccines have advanced to clinical trials.

  17. [Haemophilus influenzae b: a review on the determinants of pathogenicity and immune response to the infection].

    Science.gov (United States)

    Gómez de León, P; Cabrera-Contreras, R; Cravioto, A

    1991-01-01

    Haemophilus influenzae is still one of the main causes of diverse invasive diseases in children in México. Epidemiologic data indicate that these processes affect primarily the central nervous system and the respiratory tract. Several factors are involved in the expression of infectious disease by this organism, among them the pathogenic determinants of the parasite and those related with resistance in the host. Occurrence of disease is usually the result of the interaction between these determinants. Knowledge of these pathogenic determinants of the parasite and of factors involved in the immune response of the host have allowed an understanding of the infectious process and have directed research in a least three areas: 1) identification of bacterial membrane fractions related with diagnosis of the disease, 2) screening for immunogenic components in the bacterias as vaccine candidates to be used in the prevention of the disease and, 3) the planning of appropriate alternatives for specific antimicrobial therapy. PMID:1948428

  18. Gyrase activity and number of copies of the gyrase B subunit gene in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Cabrera-Juarez, E.; Setlow, J.K.

    1985-11-01

    Gyrase activities in extracts of various strains of Haemophilus influenzae can differ by more than an order of magnitude. Measurements of in vitro activity and copy number indicated that most of these differences arose from variations in the number of copies of the gene for the gyrase B subunit, with some strains containing multicopy plasmids coding for that subunit. The quantitative relationship between gyrase and copy number depended on the mutations in the plasmids and in the host. The possibility that the in vivo gyrase activity did not reflect the in vitro data was explored by measurement of alkaline phosphatase and ATPase activity in the extracts. Alkaline phosphatase activity increased with increasing gyrase activity measured in vitro, but ATPase activity did not. The authors conclude that extra supercoiling enhanced transcription of the alkaline phosphatase gene but not the ATPase gene and that it is unlikely that there is much discrepancy between gyrase activity assayed in vitro and the activity in the cell.

  19. Fixation and loss of hydrazine-induced premutational damage in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Kimball, R.F.; Hirsch, B.F.

    1976-01-01

    Premutational damage induced in Haemophilus influenzae by hydrazine appears to be fixed as final mutation only at replication as judged by the transformation assay. Fixation at replication is independent of the rec 1 gene, unlike the case with nitrosocarbaryl. Prior to replication premutational damage induced by hydrazine disappears by an unknown process that is not dependent on the presence of a pyrimidine dimer excision system nor on the rec 1 gene. Hydrazine does not produce detectable single-strand breaks or alkali-labile sites in the treated DNA nor gaps in DNA newly synthesized after treatment. In these respects it also differs from nitroso compounds. It is concluded that hydrazine acts to produce an altered base, possibly N(4)-aminocytosine, that produces mutations by mispairing at replication rather than by error-prone repair.

  20. Genetics and complementation of Haemophilus influenzae mutants deficient in adenosine 5'-triphosphate-dependent nuclease

    Energy Technology Data Exchange (ETDEWEB)

    Kooistra, J.; Small, G.D.; Setlow, J.K.; Shapanka, R.

    1976-04-01

    Eight different mutations in Haemophilus influenzae leading to deficiency in adenosine 5'-triphosphate (ATP)-dependent nuclease have been investigated in strains in which the mutations of the originally mutagenized strains have been transferred into the wild type. Sensitivity to mitomycin C and deoxycholate and complementation between extracts and deoxyribonucleic acid (DNA)-dependent ATPase activity have been measured. Genetic crosses have provided information on the relative position of the mutations on the genome. There are three complementation groups, corresponding to three genetic groups. The strains most sensitive to mitomycin and deoxycholate, derived from mutants originally selected on the basis of sensitivity to mitomycin C or methyl methanesulfonate, are in one group. Apparently all these sensitive strains lack DNA-dependent ATPase activity, as does a strain intermediate in sensitivity to deoxycholate, which is the sole representative of another group. There are four strains that are relatively resistant to deoxycholate and mitomycin C, and all of these contain the ATPase activity.

  1. Developing a vaccine to prevent otitis media caused by nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Khan, M Nadeem; Ren, Dabin; Kaur, Ravinder; Basha, Saleem; Zagursky, Robert; Pichichero, Michael E

    2016-07-01

    Nontypeable Haemophilus influenzae (NTHi) is a predominant organism of the upper respiratory nasopharyngeal microbiota. Its disease spectrum includes otitis media, sinusitis, non-bacteremic pneumonia and invasive infections. Protein-based vaccines to prevent NTHi infections are needed to alleviate these infections in children and vulnerable populations such as the elderly and those with chronic obstructive pulmonary disease (COPD). One NTHi protein is included in a pneumococcal conjugate vaccine and has been shown to provide efficacy. Our lab has been interested in understanding the immunogenicity of NTHi vaccine candidates P6, protein D and OMP26 for preventing acute otitis media in young children. We expect that continued investigation and progress in the development of an efficacious protein based vaccine against NTHi infections is achievable in the near future.

  2. Novobiocin resistance marker in Haemophilus influenzae that is not expressed on a plasmid

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, J.K.; McCarthy, D.; Clayton, N.L.

    1982-09-01

    The plasmid pNov2, carrying a cloned chromosomal marker conferring resistance to at least 2.5 ..mu..g of novobiocin per ml, was constructed with a new Haemophilus influenzae cloning vehicle, pDM2. The novobiocin marker of pNov2 was not normally expressed, but in Rec/sup +/ cells approximately one in 10/sup 4/ cells in a culture of a transformant became novobiocin resistant, a frequency about four orders of magnitude higher than the spontaneous mutation frequency. Variants of such cells that had lost the plasmid were also novobiocin resistant. Since Rec/sup -/ cultures bearing pNov2 showed novobiocin resistance only at the normal mutation frequency, the authors concluded that the Rec/sup +/ novobiocin-resistant transformants arose because of a rare recombination between plasmid and chromosome. Evidence is presented that novobiocin sensitivity is dominant over this particular novobiocin resistance marker.

  3. Electron microscopy of single-stranded structures in the DNA of competent Haemophilus influenzae cells

    Energy Technology Data Exchange (ETDEWEB)

    McCarthy, D.; Kupfer, D.M.

    1987-02-01

    Chromosomal DNAs from exponential-phase and competent cells of Haemophilus influenzae were examined by electron microscopy to determine whether the chromosome undergoes structural changes during competence development. Single-stranded gaps and single-stranded tails formed in chromosomal DNA during competence development. The generation of gaps was dependent on the rec-2 function. Since the rec-2 mutant is defective in the translocation of donor DNA, it was inferred that the gaps were involved in the translocation step of transformation. The generation of single-stranded tails was independent of the rec-1 and rec-2 genes. Therefore, these structures were assumed to play no direct role in the interaction of donor and recipient DNAs during transformation. Gaps were preferentially associated with a readily denaturable, possible A + T-rich fraction of the genome. This finding raised the possibility that hot spots for transformation might be associated with A + T-rich DNA.

  4. Improvement of Large-scale PRP production by Haemophilus influenzae typeb, using modified CY medium

    Directory of Open Access Journals (Sweden)

    Farshad Nojoomi

    2012-02-01

    Full Text Available Background and Objective: Haemophilus influenzae type b (Hib is a gram negative bacterium and one of the most common causative agents of acute meningitis in infants and less than 5 years old children worldwide. The production of Hib capsular polysaccharide; polyribosyl ribitolphosphate (PRP is important for the production of conjugate vaccines against Hib infections. The aim of this study is the improvement of Large-scale PRP production by Hib. Materials and Methods: Haemophilus influenzae type b standard strain ATCC10211 was cultivated in 2L fermentors contain 1.5L CY (casaminoacid yeast extract medium with normal or modified concentrations of glucose, yeast extract, hemin and NAD (nicotinamide adenine dinucleotide. Seed culture of two fermentors was inoculated to 50 L fermentor, separately and range of PRP production and Dry cell weight (DCW were studied. Results: Cultivation of Hib in 50L fermentor contained modified CY medium with 6gl-1 Glucose, 2.5 gl-1 Yeast extract, 0.03 gl-1 Hemin and 0.015 gl-1 NAD , with controlled pH at 7.3 and 30% Dissolved oxygen tension (DOT resulted to about 5.1 gl-1 DCW and 1.16 gl-1 PRP , that was significantly higher than normal CY medium. Conclusion: In conclusion, by modification in some medium components of CY medium, control of Dissolved oxygen tension and pH, the Large-scale production of PRP is improved. Improvement of PRP production leads to reduce the final cost of Hib conjugate vaccines.

  5. Rethinking the benefits and costs of childhood vaccination: the example of the Haemophilus influenzae type b vaccine.

    Science.gov (United States)

    Bärnighausen, Till; Bloom, David E; Canning, David; Friedman, Abigail; Levine, Orin S; O'Brien, Jennifer; Privor-Dumm, Lois; Walker, Damian

    2011-03-16

    Economic evaluations of health interventions, such as vaccinations, are important tools for informing health policy. Approaching the analysis from the appropriate perspective is critical to ensuring the validity of evaluation results for particular policy decisions. Using the example of cost-benefit analysis (CBA) of Haemophilus influenzae type b (Hib) vaccination, we demonstrate that past economic evaluations have mostly adopted narrow evaluation perspectives, focusing primarily on health gains, health-care cost savings, and reductions in the time costs of caring, while usually ignoring other important benefits including outcome-related productivity gains (improved economic productivity due to prevention of mental and physical disabilities), behavior-related productivity gains (economic growth due to fertility reductions as vaccination improves child survival), and community externalities (herd immunity and prevention of antibiotic resistance). We further show that potential cost reductions that could be attained through changes in the delivery of the Hib vaccine have also generally been ignored in economic evaluations. Future economic evaluations of childhood vaccinations should take full account of benefits and costs, so that policymakers have sufficient information to make well-informed decisions on vaccination implementation.

  6. Haemophilus influenzae infection drives IL-17-mediated neutrophilic allergic airways disease.

    Directory of Open Access Journals (Sweden)

    Ama-Tawiah Essilfie

    2011-10-01

    Full Text Available A subset of patients with stable asthma has prominent neutrophilic and reduced eosinophilic inflammation, which is associated with attenuated airways hyper-responsiveness (AHR. Haemophilus influenzae has been isolated from the airways of neutrophilic asthmatics; however, the nature of the association between infection and the development of neutrophilic asthma is not understood. Our aim was to investigate the effects of H. influenzae respiratory infection on the development of hallmark features of asthma in a mouse model of allergic airways disease (AAD. BALB/c mice were intraperitoneally sensitized to ovalbumin (OVA and intranasally challenged with OVA 12-15 days later to induce AAD. Mice were infected with non-typeable H. influenzae during or 10 days after sensitization, and the effects of infection on the development of key features of AAD were assessed on day 16. T-helper 17 cells were enumerated by fluorescent-activated cell sorting and depleted with anti-IL-17 neutralizing antibody. We show that infection in AAD significantly reduced eosinophilic inflammation, OVA-induced IL-5, IL-13 and IFN-γ responses and AHR; however, infection increased airway neutrophil influx in response to OVA challenge. Augmented neutrophilic inflammation correlated with increased IL-17 responses and IL-17 expressing macrophages and neutrophils (early, innate and T lymphocytes (late, adaptive in the lung. Significantly, depletion of IL-17 completely abrogated infection-induced neutrophilic inflammation during AAD. In conclusion, H. influenzae infection synergizes with AAD to induce Th17 immune responses that drive the development of neutrophilic and suppress eosinophilic inflammation during AAD. This results in a phenotype that is similar to neutrophilic asthma. Infection-induced neutrophilic inflammation in AAD is mediated by IL-17 responses.

  7. Maturation of molybdoenzymes and its influence on the pathogenesis of non-typeable Haemophilus influenzae

    Science.gov (United States)

    Dhouib, Rabeb; Pg Othman, Dk S. M.; Essilfie, Ama-Tawiah; Hansbro, Phil M.; Hanson, Jeffrey O.; McEwan, Alastair G.; Kappler, Ulrike

    2015-01-01

    Mononuclear molybdenum enzymes of the dimethylsulfoxide (DMSO) reductase family occur exclusively in prokaryotes, and a loss of some these enzymes has been linked to a loss of bacterial virulence in several cases. The MobA protein catalyzes the final step in the synthesis of the molybdenum guanine dinucleotide (MGD) cofactor that is exclusive to enzymes of the DMSO reductase family. MobA has been proposed as a potential target for control of virulence since its inhibition would affect the activities of all molybdoenzymes dependent upon MGD. Here, we have studied the phenotype of a mobA mutant of the host-adapted human pathogen Haemophilus influenzae. H. influenzae causes and contributes to a variety of acute and chronic diseases of the respiratory tract, and several enzymes of the DMSO reductase family are conserved and highly expressed in this bacterium. The mobA mutation caused a significant decrease in the activities of all Mo-enzymes present, and also resulted in a small defect in anaerobic growth. However, we did not detect a defect in in vitro biofilm formation nor in invasion and adherence to human epithelial cells in tissue culture compared to the wild-type. In a murine in vivo model, the mobA mutant showed only a mild attenuation compared to the wild-type. In summary, our data show that MobA is essential for the activities of molybdenum enzymes, but does not appear to affect the fitness of H. influenzae. These results suggest that MobA is unlikely to be a useful target for antimicrobials, at least for the purpose of treating H. influenzae infections. PMID:26594204

  8. Case of invasive nontypable Haemophilus influenzae respiratory tract infection with a large quantity of neutrophil extracellular traps in sputum

    Directory of Open Access Journals (Sweden)

    Hamaguchi S

    2012-12-01

    Full Text Available Shigeto Hamaguchi,1,* Masafumi Seki,1,* Norihisa Yamamoto,1 Tomoya Hirose,2 Naoya Matsumoto,2 Taro Irisawa,2 Ryosuke Takegawa,2 Takeshi Shimazu,2 Kazunori Tomono11Division of Infection Control and Prevention, 2Department of Traumatology and Acute Critical Medicine, Osaka University Graduate School of Medicine, Osaka, Japan *These authors contributed equally to this workAbstract: Haemophilus influenzae type b was once the most common cause of invasive H. influenzae infection, but the incidence of this disease has decreased markedly with introduction of conjugate vaccines to prevent the disease. In contrast, the incidence of invasive infection caused by nontypable H. influenzae has increased in the US and in European countries. Neutrophil extracellular traps (NETs are fibrous structures released extracellularly from activated neutrophils during inflammation, including in pneumonia, and rapidly trap and kill pathogens as a first line of immunological defense. However, their function and pathological role have not been fully investigated. Here, we report a case of fatal nontypable H. influenzae infection with severe pneumonia and bacteremia in an adult found to have a vast amount of NETs in his sputum. The patient had a two-day history of common cold-like symptoms and was taken to the emergency room as a cardiopulmonary arrest. He recovered temporarily, but died soon afterwards, although appropriate antibiotic therapy and general management had been instituted. Massive lobular pneumonia and sepsis due to nontypable H. influenzae was found, in spite of H. influenzae type b vaccine being available. His sputum showed numerous bacteria phagocytosed by neutrophils, and immunohistological staining indicated a number of NETs containing DNA, histone H3, and neutrophil elastase. This case highlights an association between formation of NETs and severe respiratory and septic infection. An increase in severe nontypable H. influenzae disease can be expected as a

  9. Role of nasopharyngeal colonization with and without bacteremia in the protection of infant rats against Haemophilus influenzae type b challenge.

    OpenAIRE

    Gilsdorf, J R; Ferrieri, P

    1985-01-01

    Nasopharyngeal colonization of infant rats with Haemophilus influenzae type b was investigated by two methods of intranasal inoculation. After traumatic instillation of the bacteria, 100% of the animals became colonized, compared with 75.5% of animals after atraumatic instillation. Among colonized rats, significantly more animals in the traumatic group developed bacteremia compared with those in the atraumatic group. Rats in the traumatic group had an onset of bacteremia at a mean of 2.6 days...

  10. The Seroepidemiology of Haemophilus influenzae Type B Prior to Introduction of an Immunization Programme in Kathmandu, Nepal

    OpenAIRE

    Marshall, Andrew S. J.; Barker, Charlotte I S; Pulickal, Anoop S.; Elizabeth Kibwana; Samir C Gautam; Elizabeth A Clutterbuck; Stephen M Thorson; Shrijana Shrestha; Neelam Adhikari; Andrew J Pollard; Kelly, Dominic F.

    2014-01-01

    Haemophilus influenzae type b (Hib) is now recognized as an important pathogen in Asia. To evaluate disease susceptibility, and as a marker of Hib transmission before routine immunization was introduced in Kathmandu, 71 participants aged 7 months-77 years were recruited and 15 cord blood samples were collected for analysis of anti-polyribosylribitol phosphate antibody levels by enzyme-linked immunosorbent assay. Only 20% of children under 5 years old had levels considered protective (>0.15 µg...

  11. Cloning and surface expression in Escherichia coli of a structural gene encoding a surface protein of Haemophilus influenzae type b.

    OpenAIRE

    Holmans, P L; Loftus, T A; Hansen, E J

    1985-01-01

    Recombinant DNA technology was used to clone a gene coding for a surface protein of Haemophilus influenzae type b (Hib) into Escherichia coli. Chromosomal DNA from a clinical isolate of Hib was cleaved with EcoRI and ligated into plasmid vectors containing three different translational reading frames. E. coli carrying recombinant plasmids were screened in a colony blot-radioimmunoassay system by using murine monoclonal antibodies (mabs) directed against cell surface-exposed proteins of Hib. m...

  12. Inhibition of human neutrophil migration in vitro by low-molecular-mass products of nontypeable Haemophilus influenzae.

    OpenAIRE

    Cundell, D R; Taylor, G W; Kanthakumar, K; Wilks, M.; Tabaqchali, S; Dorey, E; Devalia, J L; Roberts, D E; Davies, R J; Wilson, R.

    1993-01-01

    Nontypeable Haemophilus influenzae commonly causes infections in the lower and upper respiratory tract, although the mechanisms of its colonization and persistence in the airways are unclear. Culture filtrates from six clinical isolates of this bacterium were assessed for their abilities to influence neutrophil function in vitro. Each culture filtrate was assessed on six separate occasions with neutrophils obtained from six different donors. During the log and early stationary phases of growt...

  13. Clarithromycin Resistance Mechanisms of Epidemic β-Lactamase-Nonproducing Ampicillin-Resistant Haemophilus influenzae Strains in Japan.

    Science.gov (United States)

    Seyama, Shoji; Wajima, Takeaki; Nakaminami, Hidemasa; Noguchi, Norihisa

    2016-05-01

    The aim of this study was to clarify the clarithromycin resistance mechanisms of β-lactamase-nonproducing ampicillin-resistant Haemophilus influenzae strains. In all clarithromycin-resistant strains, the transcript level of acrB was significantly elevated, and these strains had a frameshift mutation in acrR Introduction of the acrR mutation into H. influenzae Rd generated a clarithromycin-resistant transformant with the same MIC as the donor strain. Our results indicate that the acrR mutation confers clarithromycin resistance by the increasing the transcription of acrB.

  14. Inhibitory effect of 1,2,4-triazole-ciprofloxacin hybrids on Haemophilus parainfluenzae and Haemophilus influenzae biofilm formation in vitro under stationary conditions.

    Science.gov (United States)

    Kosikowska, Urszula; Andrzejczuk, Sylwia; Plech, Tomasz; Malm, Anna

    2016-10-01

    Haemophilus parainfluenzae and Haemophilus influenzae, upper respiratory tract microbiota representatives, are able to colonize natural and artificial surfaces as biofilm. The aim of the present study was to assay the effect of ten 1,2,4-triazole-ciprofloxacin hybrids on planktonic or biofilm-forming haemophili cells in vitro under stationary conditions on the basis of MICs (minimal inhibitory concentrations) and MBICs (minimal biofilm inhibitory concentrations). In addition, anti-adhesive properties of these compounds were examined. The reference strains of H. parainfluenzae and H. influenzae were included. The broth microdilution microtiter plate (MTP) method with twofold dilution of the compounds, or ciprofloxacin (reference agent) in 96-well polystyrene microplates, was used. The optical density (OD) reading was made spectrophotometrically at a wavelength of 570 nm (OD570) both to measure bacterial growth and to detect biofilm-forming cells under the same conditions with 0.1% crystal violet. The following values of parameters were estimated for 1,2,4-triazole-ciprofloxacin hybrids - MIC = 0.03-15.63 mg/L, MBIC = 0.03-15.63 mg/L, MBIC/MIC = 0.125-8, depending on the compound, and for ciprofloxacin - MIC = 0.03-0.06 mg/L, MBIC = 0.03-0.12 mg/L, MBIC/MIC = 1-2. The observed strong anti-adhesive properties (95-100% inhibition) of the tested compounds were reversible during long-term incubation at subinhibitory concentrations. Thus, 1,2,4-triazole-ciprofloxacin hybrids may be considered as starting compounds for designing improved agents not only against planktonic but also against biofilm-forming Haemophilus spp. cells.

  15. Difficult identification of Haemophilus influenzae, a typical cause of upper respiratory tract infections, in the microbiological diagnostic routine.

    Science.gov (United States)

    Hinz, Rebecca; Zautner, Andreas Erich; Hagen, Ralf Matthias; Frickmann, Hagen

    2015-03-01

    Haemophilus influenzae is a key pathogen of upper respiratory tract infections. Its reliable discrimination from nonpathogenic Haemophilus spp. is necessary because merely colonizing bacteria are frequent at primarily unsterile sites. Due to close phylogenetic relationship, it is not easy to discriminate H. influenzae from the colonizer Haemophilus haemolyticus. The frequency of H. haemolyticus isolations depends on factors like sampling site, patient condition, and geographic region. Biochemical discrimination has been shown to be nonreliable. Multiplex PCR including marker genes like sodC, fucK, and hpd or sequencing of the 16S rRNA gene, the P6 gene, or multilocus-sequence-typing is more promising. For the diagnostic routine, such techniques are too expensive and laborious. If available, matrix-assisted laser-desorption-ionization time-of-flight mass spectrometry is a routine-compatible option and should be used in the first line. However, the used database should contain well-defined reference spectra, and the spectral difference between H. influenzae and H. haemolyticus is small. Fluorescence in-situ hybridization is an option for less well-equipped laboratories, but the available protocol will not lead to conclusive results in all instances. It can be used as a second line approach. Occasional ambiguous results have to be resolved by alternative molecular methods like 16S rRNA gene sequencing. PMID:25883794

  16. Non-typeable Haemophilus influenzae decreases cilia beating via protein kinase C epsilon

    Directory of Open Access Journals (Sweden)

    Bailey Kristina L

    2012-06-01

    Full Text Available Abstract Background Haemophilus influenzae infection of the nasal epithelium has long been associated with observations of decreased nasal ciliary beat frequency (CBF and injury to the ciliated epithelium. Previously, we have reported that several agents that slow CBF also have the effect of activating protein kinase C epsilon (PKCϵ activity in bronchial epithelial cells. The subsequent auto-downregulation of PKCϵ or the direct inhibition of PKCϵ leads to the specific detachment of the ciliated cells. METHODS: Primary cultures of ciliated bovine bronchial epithelial cells were exposed to filtered conditioned media supernatants from non-typeable H. influenzae (NTHi cultures. CBF and motile points were measured and PKCϵ activity assayed. Results NTHi supernatant exposure significantly and rapidly decreased CBF in a dose-dependent manner within 10 minutes of exposure. After 3 hours of exposure, the number of motile ciliated cells significantly decreased. Direct measurement of PKCϵ activity revealed a dose-dependent activation of PKCϵ in response to NTHi supernatant exposure. Both CBF and PKCϵ activity changes were only observed in fresh NTHi culture supernatant and not observed in exposures to heat-inactivated or frozen supernatants. Conclusions Our results suggest that CBF slowing observed in response to NTHi is consistent with the stimulated activation of PKCϵ. Ciliated cell detachment is associated with PKCϵ autodownregulation.

  17. Evidence for covalent attachment of phospholipid to the capsular polysaccharide of Haemophilus influenzae type b

    Energy Technology Data Exchange (ETDEWEB)

    Kuo, J.S.; Doelling, V.W.; Graveline, J.F.; McCoy, D.W.

    1985-08-01

    Cells of Haemophilus influenzae type b were grown in a liquid medium containing (TH)palmitate or ( UC)ribose or both for two generations of exponential growth. Radiolabeled type-specific capsular polysaccharide, polyribosyl ribitol phosphate (PRP), was purified from the culture supernatant by Cetavlon precipitation, ethanol fractionation, and hydroxylapatite and Sepharose 4B chromatography. The doubly labeled ( (TH)palmitate and ( UC)ribose) PRP preparation was found to coelute in a single peak from a Sepharose 4B column, suggesting that both precursors were incorporated into the purified PRP. A singly labeled ( (TH)palmitate) purified PRP preparation was found to be quantitatively immune precipitated by human serum containing antibody against PRP. Only after acid, alkaline, or phospholipase A2 treatment of PRP labeled with (TH)palmitate or (TH)palmitate and ( UC)ribose followed by chloroform-methanol extraction could most of the TH-radioactivity be recovered in the organic phase. The chloroform-soluble acid-hydrolyzed or phospholipase A2-treated product was identified as palmitic acid after thin-layer chromatography. These results strongly suggest that a phospholipid moiety is covalently associated with the H. influenzae type b polysaccharide PRP.

  18. Intranasal immunization with nontypeable Haemophilus influenzae outer membrane vesicles induces cross-protective immunity in mice.

    Directory of Open Access Journals (Sweden)

    Sandro Roier

    Full Text Available Haemophilus influenzae is a Gram-negative human-restricted bacterium that can act as a commensal and a pathogen of the respiratory tract. Especially nontypeable H. influenzae (NTHi is a major threat to public health and is responsible for several infectious diseases in humans, such as pneumonia, sinusitis, and otitis media. Additionally, NTHi strains are highly associated with exacerbations in patients suffering from chronic obstructive pulmonary disease. Currently, there is no licensed vaccine against NTHi commercially available. Thus, this study investigated the utilization of outer membrane vesicles (OMVs as a potential vaccine candidate against NTHi infections. We analyzed the immunogenic and protective properties of OMVs derived from various NTHi strains by means of nasopharyngeal immunization and colonization studies with BALB/c mice. The results presented herein demonstrate that an intranasal immunization with NTHi OMVs results in a robust and complex humoral and mucosal immune response. Immunoprecipitation revealed the most important immunogenic proteins, such as the heme utilization protein, protective surface antigen D15, heme binding protein A, and the outer membrane proteins P1, P2, P5 and P6. The induced immune response conferred not only protection against colonization with a homologous NTHi strain, which served as an OMV donor for the immunization mixtures, but also against a heterologous NTHi strain, whose OMVs were not part of the immunization mixtures. These findings indicate that OMVs derived from NTHi strains have a high potential to act as a vaccine against NTHi infections.

  19. DNA aptamers for the detection of Haemophilus influenzae type b by cell SELEX.

    Science.gov (United States)

    Bitaraf, F S; Rasooli, I; Mousavi Gargari, S L

    2016-03-01

    Haemophilus influenzae type b (Hib) causes acute bacterial meningitis (ABM) in children, with a mortality rate of about 3-6 % of the affected patients. ABM can lead to death during a period of hours to several days and, hence, rapid and early detection of the infection is crucial. Aptamers, the short single-stranded DNA or RNA with high affinity to target molecules, are selected by a high-flux screening technique known as in vitro screening and systematic evolution of ligands by exponential enrichment technology (SELEX). In this study, whole-cell SELEX was applied for the selection of target-specific aptamers with high affinity to Hib. ssDNA aptamers prepared by lambda exonuclease were incubated with the target cells (Hib). The aptameric binding rate to Hib was characterized for binding affinity after seven SELEX rounds by flow cytometry. The aptamers with higher binding affinity were cloned. Four of 68 aptamer clones were selected for sequencing. The dissociation constant (Kd) of the high-affinity aptamer clones 45 and 63 were 47.10 and 28.46 pM, respectively. These aptamers did not bind to other bacterial species, including the seven meningitis-causing bacteria. They showed distinct affinity to various H. influenzae strains only. These aptamers showed the highest affinity to Hib and the lowest affinity to H. influenzae type c and to other meningitis-causing bacteria. Clone 63 could detect Hib in patients' cerebrospinal fluid (CSF) samples at 60 colony-forming units (CFU)/mL. The results indicate applicability of the aptamers for rapid and early detection of infections brought about by Hib. PMID:26768582

  20. DNA aptamers for the detection of Haemophilus influenzae type b by cell SELEX.

    Science.gov (United States)

    Bitaraf, F S; Rasooli, I; Mousavi Gargari, S L

    2016-03-01

    Haemophilus influenzae type b (Hib) causes acute bacterial meningitis (ABM) in children, with a mortality rate of about 3-6 % of the affected patients. ABM can lead to death during a period of hours to several days and, hence, rapid and early detection of the infection is crucial. Aptamers, the short single-stranded DNA or RNA with high affinity to target molecules, are selected by a high-flux screening technique known as in vitro screening and systematic evolution of ligands by exponential enrichment technology (SELEX). In this study, whole-cell SELEX was applied for the selection of target-specific aptamers with high affinity to Hib. ssDNA aptamers prepared by lambda exonuclease were incubated with the target cells (Hib). The aptameric binding rate to Hib was characterized for binding affinity after seven SELEX rounds by flow cytometry. The aptamers with higher binding affinity were cloned. Four of 68 aptamer clones were selected for sequencing. The dissociation constant (Kd) of the high-affinity aptamer clones 45 and 63 were 47.10 and 28.46 pM, respectively. These aptamers did not bind to other bacterial species, including the seven meningitis-causing bacteria. They showed distinct affinity to various H. influenzae strains only. These aptamers showed the highest affinity to Hib and the lowest affinity to H. influenzae type c and to other meningitis-causing bacteria. Clone 63 could detect Hib in patients' cerebrospinal fluid (CSF) samples at 60 colony-forming units (CFU)/mL. The results indicate applicability of the aptamers for rapid and early detection of infections brought about by Hib.

  1. A novel nickel responsive MerR-like regulator, NimR, from Haemophilus influenzae.

    Science.gov (United States)

    Kidd, Stephen P; Djoko, Karrera Y; Ng, JiaQi; Argente, M Pilar; Jennings, Michael P; McEwan, Alastair G

    2011-10-01

    We have identified a novel regulator from the MerR family of transcription factors in the bacterial pathogen Haemophilus influenzae (HI1623; nickel-associated merR-like Regulator--NimR). NimR regulates the expression of a Ni(2+) uptake transporter (NikKLMQO). The promoters for nimR and the nik operon are divergent and overlapping and NimR binds at a site between the promoter elements for nikKLMQO. Expression of this operon requires NimR and depends on Ni(2+). Growth rates of the H. influenzae nimR and nikQ mutants were reduced in chemically defined media compared to the wild type and the mutants were unable to grow in the presence of EDTA. The mutant strains were less tolerant of acidic pH and the wild type Rd KW20 could not tolerate low pH in the presence of fluoramide, a urease specific inhibitor, confirming that both nickel transport and urea hydrolysis are a central process in pH control. H. influenzae nimR and nikQ strains were deficient in urease activity, but this could be specifically restored by the addition of excess Ni(2+). NimR did not directly regulate the expression of urease genes but the activity of urease requires both nimR and nikQ. Purified NimR is a dimer that binds 1 Ni(2+)ion. NimR is the first example of a Ni-dependent regulator from the MerR family and targeting a metal ion uptake system; it is distinct from NikR the Ni-responsive regulators of the ribbon-helix-helix family. PMID:21952667

  2. Laboratory characterization of invasive Haemophilus influenzae isolates from Nunavut, Canada, 2000–2012

    Science.gov (United States)

    Tsang, Raymond S. W.; Li, Y. Anita; Mullen, Angie; Baikie, Maureen; Whyte, Kathleen; Shuel, Michelle; Tyrrell, Gregory; Rotondo, Jenny A. L.; Desai, Shalini; Spika, John

    2016-01-01

    Background With invasive Haemophilus influenzae serotype b (Hib) disease controlled by vaccination with conjugate Hib vaccines, there is concern that invasive disease due to non-serotype b strains may emerge. Objective This study characterized invasive H. influenzae (Hi) isolates from Nunavut, Canada, in the post-Hib vaccine era. Methods Invasive H. influenzae isolates were identified by conventional methods at local hospitals; and further characterized at the provincial and federal public health laboratories, including detection of serotype antigens and genes, multi-locus sequence typing and antibiotic susceptibility. Results Of the 89 invasive H. influenzae cases identified from 2000 to 2012, 71 case isolates were available for study. There were 43 serotype a (Hia), 12 Hib, 2 Hic, 1 Hid, 1 Hie, 2 Hif and 10 were non-typeable (NT). All 43 Hia were biotype II, sequence type (ST)-23. Three related STs were found among the Hib isolates: ST-95 (n=9), ST-635 (n=2) and ST-44 (n=1). Both Hif belonged to ST-124 and the 2 Hic were typed as ST-9. The remaining Hid (ST-1288) and Hie (ST-18) belonged to 2 separate clones. Of the 10 NT strains, 3 were typed as ST-23 and the remaining 7 isolates each belonged to a unique ST. Eight Hib and 1 NT-Hi were found to be resistant to ampicillin due to β-lactamase production. No resistance to other antibiotics was detected. Conclusion During the period of 2000–2012, Hia was the predominant serotype causing invasive disease in Nunavut. This presents a public health concern due to an emerging clone of Hia as a cause of invasive H. influenzae disease and the lack of published guidelines for the prophylaxis of contacts. The clonal nature of Hia could be the result of spread within an isolated population, and/or unique characteristics of this strain to cause invasive disease. Further study of Hia in other populations may provide important information on this emerging pathogen. No antibiotic resistance was detected among Hia isolates; a

  3. Progress of Haemophilus influenzae type a and related vaccine%a 型流感嗜血杆菌及其疫苗的研究进展

    Institute of Scientific and Technical Information of China (English)

    苗鑫(综述); 谢贵林; 赵志强(审校)

    2016-01-01

    随着b型流感嗜血杆菌(Haemophilus influenzae type b, Hib)结合疫苗在全球的广泛应用,Hib相关疾病的发病率明显下降;a型流感嗜血杆菌( Haemophilus influenzae type a, Hia)逐渐成为侵袭性流感嗜血杆菌疾病的重要病原体,且因其发病率高、临床症状重、死亡率高而日益受到关注。本文从Hia的流行概况、相关疾病、药物敏感性、易感人群及其疫苗研究作一综述。%The morbidity of Haemophilus influenzae type b( Hib) related diseases has reduced substantially,since Hib conju-gate vaccines were widely used all over the world.However,Haemophilus influenzae type a ( Hia) has become an important pathogen that causes invasive Haemophilus influenzae diseases, and been concerned with a high incidence for the related diseases, severe clinical symptoms and a high mortality, increasingly.We review epidemiology, related diseases, antibiotic susceptibility, susceptible population and related vaccine research of Hia diseases.

  4. Effect of Fluoroquinolones and Macrolides on Eradication and Resistance of Haemophilus influenzae in Chronic Obstructive Pulmonary Disease.

    Science.gov (United States)

    Pettigrew, Melinda M; Tsuji, Brian T; Gent, Janneane F; Kong, Yong; Holden, Patricia N; Sethi, Sanjay; Murphy, Timothy F

    2016-07-01

    Little is known about the effect of antibiotics on eradication of carriage and development of resistance in Haemophilus influenzae in individuals with chronic obstructive pulmonary disease (COPD). Our goals were to assess antibiotic susceptibilities, prevalence of resistance genes, and development of resistance in H. influenzae and to evaluate the effect of macrolide and fluoroquinolone administration on H. influenzae eradication. Data were from a 15-year longitudinal study of COPD. Genome sequence data were used to determine genotype and identify resistance genes. MICs of antibiotics were determined by reference broth microdilution. Generalized linear mixed models were used to evaluate associations between antibiotic use and H. influenzae eradication. We examined 267 H. influenzae isolates from 77 individuals. All newly acquired H. influenzae isolates were susceptible to azithromycin. Five of 27 (19%) strains developed 4-fold increases in azithromycin MICs and reached or exceeded the susceptibility breakpoint (≤4 μg/ml) during exposure. H. influenzae isolates were uniformly susceptible to ciprofloxacin, levofloxacin, and moxifloxacin (MIC90s of 0.015, 0.015, and 0.06, respectively); there were no mutations in quinolone resistance-determining regions. Fluoroquinolone administration was associated with increased H. influenzae eradication compared to macrolides (odds ratio [OR], 16.67; 95% confidence interval [CI], 2.67 to 104.09). There was no difference in H. influenzae eradication when comparing macrolide administration to no antibiotic (OR, 1.89; 95% CI, 0.43 to 8.30). Fluoroquinolones are effective in eradicating H. influenzae in individuals with COPD. Macrolides are ineffective in eradicating H. influenzae, and their use in COPD patients may lead to decreased macrolide susceptibility and resistance.

  5. Tendência das meningites por Haemophilus influenzae tipo b no Brasil, em menores de 5 anos, no período de 1983 a 2002 Trends in Haemophilus influenzae type b meningitis in Brazil in children under five years of age from 1983 through 2002

    Directory of Open Access Journals (Sweden)

    Sybelle de Souza Castro Miranzi

    2006-10-01

    Full Text Available Trata-se de um estudo ecológico, tipo série histórica (1983-2002, onde foram calculados os coeficientes de incidência, mortalidade e letalidade de meningites por Haemophilus influenzae , tipo b, no Brasil, e avaliou-se a tendência da morbi-mortalidade em menores de 5 anos. Para a análise de tendência dos coeficientes construíram-se modelos de regressão polinomial para as faixas etárias de The study was based on an ecological design using a historical time series (1983-2002, related to Haemophilus influenzae type b meningitis in Brazil. Incidence, mortality and case-fatality rates, as well as trends in incidence and morbidity-mortality were estimated in children less than 5 years of age. Polynomial regression analysis was used to analyze trends, adopting a significance level of 0.05. 43.9% of confirmed cases occurred in infants less than 1 year old and 38.7% in children 1-4 years old. The observed rates were also highest in these two age strata. The incidence and mortality rates showed an increasing trend, until approximately 1999, when a quick decline was observed. The study results reinforce the effectiveness of the Vaccination Program against HIB in Brazil, which benefited age ranges that did not receive the vaccine (Herd Immunity.

  6. Detection of Haemophilus influenzae in respiratory secretions from pneumonia patients by quantitative real-time polymerase chain reaction.

    Science.gov (United States)

    Abdeldaim, Guma M K; Strålin, Kristoffer; Kirsebom, Leif A; Olcén, Per; Blomberg, Jonas; Herrmann, Björn

    2009-08-01

    A quantitative real-time polymerase chain reaction (PCR) based on the omp P6 gene was developed to detect Haemophilus influenzae. Its specificity was determined by analysis of 29 strains of 11 different Haemophilus spp. and was compared with PCR assays having other target genes: rnpB, 16S rRNA, and bexA. The method was evaluated on nasopharyngeal aspirates from 166 adult patients with community-acquired pneumonia. When 10(4) DNA copies/mL was used as cutoff limit for the method, P6 PCR had a sensitivity of 97.5% and a specificity of 96.0% compared with the culture. Of 20 culture-negative but P6 PCR-positive cases, 18 were confirmed by fucK PCR as H. influenzae. Five (5.9%) of 84 nasopharyngeal aspirates from adult controls tested PCR positive. We conclude that the P6 real-time PCR is both sensitive and specific for identification of H. influenzae in respiratory secretions. Quantification facilitates discrimination between disease-causing H. influenzae strains and commensal colonization. PMID:19446978

  7. In vitro activity of telithromycin against Haemophilus influenzae at epithelial lining fluid concentrations

    Directory of Open Access Journals (Sweden)

    Drago Lorenzo

    2008-01-01

    Full Text Available Abstract Background Haemophilus influenzae is one of the main aetiological agents of community-acquired respiratory tract infections. The primary aim of this study was to evaluate the antibacterial activity of telithromycin against H. influenzae clinical isolates showing different pattern of resistance in comparison with azithromycin and clarithromycin at 1/4 ×, 1/2 ×, 1 ×, 2 ×, 4 × minimum inhibitory concentration (MIC and to peak concentrations in epithelial lining fluid (ELF. The secondary aim was to determine the influence of CO2 enriched atmosphere on bacterial susceptibility. Results Telithromycin showed high activity against H. influenzae, including strains susceptible to β-lactams (n = 200, β-lactamase producer (n = 50 and β-lactamase negative ampicillin resistant (BLNAR (n = 10, with MIC from ≤0.03 to 4 mg/L, and MIC50/MIC90 of 1/2 mg/L with susceptibility rate of 100%, and minimum bactericidal concentrations (MBC from 2 to 4-fold higher than the MIC. Azithromycin was the most active tested macrolide (range: 0.25 – 4 mg/L; MIC50/MIC90: 1/2 mg/L, comparable to telithromycin, while clarithromycin showed the highest MICs and MBCs (range: 0.25 – 8 mg/L; MIC50/MIC90: 2/8 mg/L. In time-kill studies, telithromycin showed a bactericidal activity at the higher concentrations (4 – 2 × MIC and ELF against all the strains, being complete after 12 – 24 hours from drug exposition. At MIC concentrations, at ambient air, bactericidal activity of telithromycin and azithromycin was quite similar at 12 hours, and better than that of clarithromycin. Besides, telithromycin and clarithromycin at ELF concentrations were bactericidal after 12 hours of incubation for most strains, while 24 hours were needed to azithromycin to be bactericidal. Incubation in CO2 significantly influenced the MICs and MBCs, and only slightly the in vitro killing curves. Conclusion Telithromycin showed an in-vitro potency against H. influenzae comparable to

  8. Characterization of the rec-1 gene of Haemophilus influenzae and behavior of the gene in Escherichia coli

    International Nuclear Information System (INIS)

    The rec-1 gene of Haemophilus influenzae was cloned into a shuttle vector that replicates in Escherichia coli as well as in H. influenzae. The plasmid, called pRec1, complemented the defects of a rec-1 mutant in repair of UV damage, transformation, and ability of prophage to be induced by UV radiation. Although UV resistance and recombination were caused by pRec1 in E. coli recA mutants, UV induction of lambda and UV mutagenesis were not. We suggest that the ability of the H. influenzae Rec-1 protein to cause cleavage of repressors but not the recombinase function differs from that of the E. coli RecA protein

  9. Characterization of the rec-1 gene of Haemophilus influenzae and behavior of the gene in Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, J.K.; Spikes, D.; Griffin, K.

    1988-09-01

    The rec-1 gene of Haemophilus influenzae was cloned into a shuttle vector that replicates in Escherichia coli as well as in H. influenzae. The plasmid, called pRec1, complemented the defects of a rec-1 mutant in repair of UV damage, transformation, and ability of prophage to be induced by UV radiation. Although UV resistance and recombination were caused by pRec1 in E. coli recA mutants, UV induction of lambda and UV mutagenesis were not. We suggest that the ability of the H. influenzae Rec-1 protein to cause cleavage of repressors but not the recombinase function differs from that of the E. coli RecA protein.

  10. Comparison of radiometric and conventional culture systems in detecting Haemophilus influenzae type b bacteremia in rats

    International Nuclear Information System (INIS)

    To compare the efficiency of detecting Haemophilus influenzae type b bacteremia by the BACTEC radiometric system and a conventional Trypticase soy broth blood culture system, the authors developed an in vivo model of bacteremia in rats. After intravenous injection of 50 to 200 CFU into adult rats, there was a linear logarithmic increase in CFU per milliliter of rat blood during the first 10 h (r = 0.98), allowing accurate prediction of the level of bacteremia with time. Culture bottles were inoculated with 0.5 ml of blood obtained by cardiac puncture and processed as clinical samples in the microbiology laboratory with RS and conventional protocols. They found the following. (i) The first detection of bacteremia by RS was similar to that by TSB if a Gram stain of the TSB was done on day 1 and was superior if that smear was omitted (P less than 0.01). (ii) The detection times in both systems were comparable at different magnitudes of bacteremia (10(1) to 10(4) CFU/ml). (iii) Supplementation of inoculated bottles with 2 ml of sterile rat blood interfered with Gram stain detection in TSB but resulted in increased 14CO2 production in RS. (iv) No difference in detection time was found between RS and TSB for four different clinical isolates. These studies show that, in a biologically relevant model, the detection of positive blood cultures for H. influenzae type b by RS was comparable to or better than detection by TSB when blood was processed analogously to clinical specimens

  11. Haemophilus influenzae: Aðgreining frá Haemophilus haemolyticus og hjúpgreining með multiplex PCR aðferð

    OpenAIRE

    Jana Birta Björnsdóttir 1989

    2013-01-01

    Bakteríur af ættkvíslinni Haemophilus eru Gram neikvæðir stafir og finnast helst í efri loftvegum manna. Þær eiga það sameiginlegt að þurfa hemin, niðurbrotsefni blóðs til að vaxa. Haemophilus influenzae (Hi) er algengust þeirra til að valda sýkingum. Hi er annaðhvort með eða án fjölsykruhjúps. Hjúpgerðir eru flokkaðar eftir ónæmisfræðilegum mismun í byggingu fjölsykranna. Alls hefur verið lýst sex hjúpgerðum en þær nefnast a, b, c, d, e og f. Áður en bólusetningar gegn hjúpgerð b (Hib) hófus...

  12. Study on Haemophilus influenzae type b diseases in China: the past, present and future.

    Science.gov (United States)

    Yang, Y; Shen, X; Jiang, Z; Liu, X; Leng, Z; Lu, D; Rao, J; Liu, J; Chang, L

    1998-09-01

    Meningitis caused by Haemophilus influenzae type b (Hib) is a common and serious disease for which there now are WHO-certified vaccines that are recommended for universal infant immunization in North America and European countries. If these vaccines are to be recommended in Asia, it is necessary to know the incidence, age distribution and clinical outcome of Hib meningitis and other systemic infections in this region. Data on Hib disease in China are scanty. Hib meningitis was common during the 1950s in China, accounting for up to 16% of all of pyogenic meningitis (up to 38% of cases were caused by unknown pathogens), despite severe epidemics of meningococcal meningitis during that period. Since 1989 we have conducted hospital- and community-based etiologic and epidemiologic studies of bacterial meningitis. Hib accounts for 30 to 50% of bacterial meningitis in China. The incidence of Hib meningitis in Hefei City was 10.4 per 100000 children death rate of children by one-third by the year 2000, greater efforts should be made to reduce the mortality of children with pneumonia. Our preliminary study showed that about one-fourth to one-third of cases of pneumonia in Chinese children might be caused by Hib. Therefore Hib vaccination for infants and children in China might be an effective and valuable procedure to achieve the goal.

  13. Nontypeable Haemophilus influenzae Induces Sustained Lung Oxidative Stress and Protease Expression

    Science.gov (United States)

    King, Paul T.; Sharma, Roleen; O’Sullivan, Kim; Selemidis, Stavros; Lim, Steven; Radhakrishna, Naghmeh; Lo, Camden; Prasad, Jyotika; Callaghan, Judy; McLaughlin, Peter; Farmer, Michael; Steinfort, Daniel; Jennings, Barton; Ngui, James; Broughton, Bradley R. S.; Thomas, Belinda; Essilfie, Ama-Tawiah; Hickey, Michael; Holmes, Peter W.; Hansbro, Philip; Bardin, Philip G.; Holdsworth, Stephen R.

    2015-01-01

    Nontypeable Haemophilus influenzae (NTHi) is a prevalent bacterium found in a variety of chronic respiratory diseases. The role of this bacterium in the pathogenesis of lung inflammation is not well defined. In this study we examined the effect of NTHi on two important lung inflammatory processes 1), oxidative stress and 2), protease expression. Bronchoalveolar macrophages were obtained from 121 human subjects, blood neutrophils from 15 subjects, and human-lung fibroblast and epithelial cell lines from 16 subjects. Cells were stimulated with NTHi to measure the effect on reactive oxygen species (ROS) production and extracellular trap formation. We also measured the production of the oxidant, 3-nitrotyrosine (3-NT) in the lungs of mice infected with this bacterium. NTHi induced widespread production of 3-NT in mouse lungs. This bacterium induced significantly increased ROS production in human fibroblasts, epithelial cells, macrophages and neutrophils; with the highest levels in the phagocytic cells. In human macrophages NTHi caused a sustained, extracellular production of ROS that increased over time. The production of ROS was associated with the formation of macrophage extracellular trap-like structures which co-expressed the protease metalloproteinase-12. The formation of the macrophage extracellular trap-like structures was markedly inhibited by the addition of DNase. In this study we have demonstrated that NTHi induces lung oxidative stress with macrophage extracellular trap formation and associated protease expression. DNase inhibited the formation of extracellular traps. PMID:25793977

  14. Nontypable Haemophilus influenzae displays a prevalent surface structure molecular pattern in clinical isolates.

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    Pau Martí-Lliteras

    Full Text Available Non-typable Haemophilus influenzae (NTHi is a gram negative pathogen that causes acute respiratory infections and is associated with the progression of chronic respiratory diseases. Previous studies have established the existence of a remarkable genetic variability among NTHi strains. In this study we show that, in spite of a high level of genetic heterogeneity, NTHi clinical isolates display a prevalent molecular feature, which could confer fitness during infectious processes. A total of 111 non-isogenic NTHi strains from an identical number of patients, isolated in two distinct geographical locations in the same period of time, were used to analyse nine genes encoding bacterial surface molecules, and revealed the existence of one highly prevalent molecular pattern (lgtF+, lic2A+, lic1D+, lic3A+, lic3B+, siaA-, lic2C+, ompP5+, oapA+ displayed by 94.6% of isolates. Such a genetic profile was associated with a higher bacterial resistance to serum mediated killing and enhanced adherence to human respiratory epithelial cells.

  15. Development and technology transfer of Haemophilus influenzae type b conjugate vaccines for developing countries.

    Science.gov (United States)

    Beurret, Michel; Hamidi, Ahd; Kreeftenberg, Hans

    2012-07-13

    This paper describes the development of a Haemophilus influenzae type b (Hib) conjugate vaccine at the National Institute for Public Health and the Environment/Netherlands Vaccine Institute (RIVM/NVI, Bilthoven, The Netherlands), and the subsequent transfer of its production process to manufacturers in developing countries. In 1998, at the outset of the project, the majority of the world's children were not immunized against Hib because of the high price and limited supply of the conjugate vaccines, due partly to the fact that local manufacturers in developing countries did not master the Hib conjugate production technology. To address this problem, the RIVM/NVI has developed a robust Hib conjugate vaccine production process based on a proven model, and transferred this technology to several partners in India, Indonesia, Korea and China. As a result, emerging manufacturers in developing countries acquired modern technologies previously unavailable to them. This has in turn facilitated their approach to producing other conjugate vaccines. As an additional spin-off from the project, a World Health Organization (WHO) Hib quality control (QC) course was designed and conducted at the RIVM/NVI, resulting in an increased regulatory capacity for conjugate vaccines in developing countries at the National Regulatory Authority (NRA) level. For the local populations, this has translated into an increased and sustainable supply of affordable Hib conjugate-containing combination vaccines. During the course of this project, developing countries have demonstrated their ability to produce large quantities of high-quality modern vaccines after a successful transfer of the technology.

  16. Peptides complementary to the active loop of porin P2 from Haemophilus influenzae modulate its activity

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    Galdiero S

    2012-05-01

    Full Text Available Marco Cantisani,1 Mariateresa Vitiello,2 Annarita Falanga,1 Emiliana Finamore,2 Marilena Galdiero,2 Stefania Galdiero11Department of Biological Sciences, CIRPeB and IBB CNR, University of Naples "Federico II," Napoli, Italy; 2Department of Experimental Medicine, II University of Naples, Napoli, ItalyAbstract: Haemophilus influenzae type b (Hib is one of the leading causes of invasive bacterial infection in young children. It is characterized by inflammation that is mainly mediated by cytokines and chemokines. One of the most abundant components of the Hib outer membrane is the P2 porin, which has been shown to induce the release of several inflammatory cytokines. A synthetic peptide corresponding to loop L7 of the porin activates JNK and p38 mitogen-activated protein kinase (MAPK pathways. We report a novel use of the complementary peptide approach to design a peptide that is able to bind selectively to the protein P2, thereby reducing its activity. This work provides insights into essential molecular details of P2 that may affect the pathogenesis of Hib infections where interruption of the signaling cascade could represent an attractive therapeutic strategy.Keywords: complementary-peptide, rational design, porin

  17. A biphasic epigenetic switch controls immunoevasion, virulence and niche adaptation in non-typeable Haemophilus influenzae.

    Science.gov (United States)

    Atack, John M; Srikhanta, Yogitha N; Fox, Kate L; Jurcisek, Joseph A; Brockman, Kenneth L; Clark, Tyson A; Boitano, Matthew; Power, Peter M; Jen, Freda E-C; McEwan, Alastair G; Grimmond, Sean M; Smith, Arnold L; Barenkamp, Stephen J; Korlach, Jonas; Bakaletz, Lauren O; Jennings, Michael P

    2015-01-01

    Non-typeable Haemophilus influenzae contains an N(6)-adenine DNA-methyltransferase (ModA) that is subject to phase-variable expression (random ON/OFF switching). Five modA alleles, modA2, modA4, modA5, modA9 and modA10, account for over two-thirds of clinical otitis media isolates surveyed. Here, we use single molecule, real-time (SMRT) methylome analysis to identify the DNA-recognition motifs for all five of these modA alleles. Phase variation of these alleles regulates multiple proteins including vaccine candidates, and key virulence phenotypes such as antibiotic resistance (modA2, modA5, modA10), biofilm formation (modA2) and immunoevasion (modA4). Analyses of a modA2 strain in the chinchilla model of otitis media show a clear selection for ON switching of modA2 in the middle ear. Our results indicate that a biphasic epigenetic switch can control bacterial virulence, immunoevasion and niche adaptation in an animal model system. PMID:26215614

  18. A biphasic epigenetic switch controls immunoevasion, virulence and niche adaptation in non-typeable Haemophilus influenzae.

    Science.gov (United States)

    Atack, John M; Srikhanta, Yogitha N; Fox, Kate L; Jurcisek, Joseph A; Brockman, Kenneth L; Clark, Tyson A; Boitano, Matthew; Power, Peter M; Jen, Freda E-C; McEwan, Alastair G; Grimmond, Sean M; Smith, Arnold L; Barenkamp, Stephen J; Korlach, Jonas; Bakaletz, Lauren O; Jennings, Michael P

    2015-01-01

    Non-typeable Haemophilus influenzae contains an N(6)-adenine DNA-methyltransferase (ModA) that is subject to phase-variable expression (random ON/OFF switching). Five modA alleles, modA2, modA4, modA5, modA9 and modA10, account for over two-thirds of clinical otitis media isolates surveyed. Here, we use single molecule, real-time (SMRT) methylome analysis to identify the DNA-recognition motifs for all five of these modA alleles. Phase variation of these alleles regulates multiple proteins including vaccine candidates, and key virulence phenotypes such as antibiotic resistance (modA2, modA5, modA10), biofilm formation (modA2) and immunoevasion (modA4). Analyses of a modA2 strain in the chinchilla model of otitis media show a clear selection for ON switching of modA2 in the middle ear. Our results indicate that a biphasic epigenetic switch can control bacterial virulence, immunoevasion and niche adaptation in an animal model system.

  19. Modelling the impact of vaccination on curtailing Haemophilus influenzae serotype 'a'.

    Science.gov (United States)

    Konini, Angjelina; Moghadas, Seyed M

    2015-12-21

    Haemophilus influenzae serotype a (Hia) is a human-restricted bacterial pathogen transmitted via direct contacts with an infectious individual. Currently, there is no vaccine available for prevention of Hia, and the disease is treated with antibiotics upon diagnosis. With ongoing efforts for the development of an anti-Hia protein-polysaccharide conjugated vaccine, we sought to investigate the effect of vaccination on curtailing Hia infection. We present the first stochastic model of Hia transmission and control dynamics, and parameterize it using available estimates in the literature. Since both naturally acquired and vaccine-induced immunity wane with time, model simulations show three important results. First, vaccination of only newborns cannot eliminate the pathogen from the population, even when a booster program is implemented with a high coverage. Second, achieving and maintaining a sufficiently high level of herd immunity for pathogen elimination requires vaccination of susceptible individuals in addition to a high vaccination coverage of newborns. Third, for a low vaccination rate of susceptible individuals, a high coverage of booster dose may be needed to raise the level of herd immunity for Hia eradication. Our findings highlight the importance of vaccination and timely boosting of the individual׳s immunity within the expected duration of vaccine-induced protection against Hia. When an anti-Hia vaccine becomes available, enhanced surveillance of Hia incidence and herd immunity could help determine vaccination rates and timelines for booster doses necessary to eliminate Hia from affected populations.

  20. Extensive cotransformation of natural variation into chromosomes of naturally competent Haemophilus influenzae.

    Science.gov (United States)

    Mell, Joshua Chang; Lee, Jae Yun; Firme, Marlo; Sinha, Sunita; Redfield, Rosemary J

    2014-04-01

    Naturally competent bacterial species actively take up environmental DNA and can incorporate it into their chromosomes by homologous recombination. This can bring genetic variation from environmental DNA to recipient chromosomes, often in multiple long "donor" segments. Here, we report the results of genome sequencing 96 colonies of a laboratory Haemophilus influenzae strain, which had been experimentally transformed by DNA from a diverged clinical isolate. Donor segments averaged 6.9 kb (spanning several genes) and were clustered into recombination tracts of ~19.5 kb. Individual colonies had replaced from 0.1 to 3.2% of their chromosomes, and ~1/3 of all donor-specific single-nucleotide variants were present in at least one recombinant. We found that nucleotide divergence did not obviously limit the locations of recombination tracts, although there were small but significant reductions in divergence at recombination breakpoints. Although indels occasionally transformed as parts of longer recombination tracts, they were common at breakpoints, suggesting that indels typically block progression of strand exchange. Some colonies had recombination tracts in which variant positions contained mixtures of both donor and recipient alleles. These tracts were clustered around the origin of replication and were interpreted as the result of heteroduplex segregation in the original transformed cell. Finally, a pilot experiment demonstrated the utility of natural transformation for genetically dissecting natural phenotypic variation. We discuss our results in the context of the potential to merge experimental and population genetic approaches, giving a more holistic understanding of bacterial gene transfer. PMID:24569039

  1. Nontypeable Haemophilus influenzae in chronic obstructive pulmonary disease and lung cancer

    Directory of Open Access Journals (Sweden)

    Seyed Javad Moghaddam

    2011-01-01

    Full Text Available Seyed Javad Moghaddam1, Cesar E Ochoa1,2, Sanjay Sethi3, Burton F Dickey1,41Department of Pulmonary Medicine, the University of Texas MD Anderson Cancer Center, Houston, TX, USA; 2Tecnológico de Monterrey School of Medicine, Monterrey, Nuevo León, Mexico; 3Department of Medicine, University at Buffalo, State University of New York, Buffalo, NY, USA; 4Center for Inflammation and Infection, Institute of Biosciences and Technology, Texas A&M Health Science Center, Houston, TX, USAAbstract: Chronic obstructive pulmonary disease (COPD is predicted to become the third leading cause of death in the world by 2020. It is characterized by airflow limitation that is not fully reversible. The airflow limitation is usually progressive and associated with an abnormal inflammatory response of the lungs to noxious particles and gases, most commonly cigarette smoke. Among smokers with COPD, even following withdrawal of cigarette smoke, inflammation persists and lung function continues to deteriorate. One possible explanation is that bacterial colonization of smoke-damaged airways, most commonly with nontypeable Haemophilus influenzae (NTHi, perpetuates airway injury and inflammation. Furthermore, COPD has also been identified as an independent risk factor for lung cancer irrespective of concomitant cigarette smoke exposure. In this article, we review the role of NTHi in airway inflammation that may lead to COPD progression and lung cancer promotion.Keywords: COPD, NTHi, inflammation

  2. A biphasic epigenetic switch controls immunoevasion, virulence and niche adaptation in non-typeable Haemophilus influenzae

    Science.gov (United States)

    Atack, John M.; Srikhanta, Yogitha N.; Fox, Kate L.; Jurcisek, Joseph A.; Brockman, Kenneth L.; Clark, Tyson A.; Boitano, Matthew; Power, Peter M.; Jen, Freda E.-C.; McEwan, Alastair G.; Grimmond, Sean M.; Smith, Arnold L.; Barenkamp, Stephen J.; Korlach, Jonas; Bakaletz, Lauren O.; Jennings, Michael P.

    2015-01-01

    Non-typeable Haemophilus influenzae contains an N6-adenine DNA-methyltransferase (ModA) that is subject to phase-variable expression (random ON/OFF switching). Five modA alleles, modA2, modA4, modA5, modA9 and modA10, account for over two-thirds of clinical otitis media isolates surveyed. Here, we use single molecule, real-time (SMRT) methylome analysis to identify the DNA-recognition motifs for all five of these modA alleles. Phase variation of these alleles regulates multiple proteins including vaccine candidates, and key virulence phenotypes such as antibiotic resistance (modA2, modA5, modA10), biofilm formation (modA2) and immunoevasion (modA4). Analyses of a modA2 strain in the chinchilla model of otitis media show a clear selection for ON switching of modA2 in the middle ear. Our results indicate that a biphasic epigenetic switch can control bacterial virulence, immunoevasion and niche adaptation in an animal model system. PMID:26215614

  3. Attempts to induce mutations in Haemophilus influenzae with the base analogues 5-bromodeoxyuridine and 2-aminopurine

    Energy Technology Data Exchange (ETDEWEB)

    Kimball, R.F.; Perdue, S.W.

    1977-01-01

    Attempts were made to induce mutations in Haemophilus influenzae with the base analogues 5-bromodeoxyuridine and 2-aminopurine. These attempts were unsuccessful. Incorporation studies with BrdUrd showed, in agreement with earlier studies on Escherichia coli, that BrdUrd was discriminated against when dThd was also present but was incorporated to essentially the same extent as dThd when only BrdUrd was present. In this latter case, strands fully substituted with BrdUrd were produced, but survival data suggest that bacteria deriving their DNA by replication on such fully substituted templates were inviable. However, bacteria with about 20% of the thymine substituted with bromouracil were usually viable. No mutations could be detected in the descendants of such bacteria. The reasons for this are discussed and it is concluded that in all probability the replication system in species rarely if ever treats incorporated bromouracil as anything except a thymine analogue. The alternative possibility, that the negative results are a consequence of the absence of the reclex (SOS) error-prone repair system in this species, is considered much less likely.

  4. Properties of mutants of haemophilus influenzae deficient in ATP-dependent deoxyribonuclease

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, J.K.

    1976-01-01

    Eight isogenic Haemophilus influenzae strains whose extracts lack ATP-dependent deoxyribonuclease activity (Add/sup -/ mutants) form three complementation and genetic linkage groups. Since there are known to be three subunits of the enzyme, these data suggest that each of the three genes specifies a different subunit. Gel electrophoresis of partially purified mutant extracts indicates that the smallest subunit is missing in one of the groups but is present in all the other mutants. The mutants are more sensitive to a variety of chemical agents than the wild type. The most sensitive mutants lack the ATPase activity associated with the enzyme. These strains exhibit aberrant incorporation of tritiated thymidine, which starts up more rapidly and shuts off sooner than in the wild type. An extracellular compound is responsible for most of this effect, in that wild type cells put into medium in which Add/sup -/ cells have been growing show a similar aberrant incorporation. The effect of these media can be mimicked by cyclic AMP and cyclic GMP, although millimolar concentrations are required. It is postulated that the Add/sup -/ mutants are more permeable to many substances than the wild type, partly because of the extracellular compound usually surrounding them, and the increased permeability might be responsible for the mutants' nonviability.

  5. Streptococcus pneumoniae synergizes with nontypeable Haemophilus influenzae to induce inflammation via upregulating TLR2

    Directory of Open Access Journals (Sweden)

    Kweon Soo-Mi

    2008-07-01

    Full Text Available Abstract Background Toll-like receptor 2 (TLR2 plays a critical role in mediating inflammatory/immune responses against bacterial pathogens in lung. Streptococcus pneumoniae (S. pneumoniae and nontypeable Haemophilus influenzae (NTHi were previously reported to synergize with each other to induce inflammatory responses. Despite the relatively known intracellular signaling pathways involved in the synergistic induction of inflammation, it is still unclear if both bacterial pathogens also synergistically induce expression of surface TLR2. Results Here we provide direct evidence that S. pneumoniae synergizes with NTHi to upregulate TLR2 expression in lung and middle ear of the mice. Pneumolysin (PLY appears to be the major virulence factor involved in this synergism. Moreover, S. pneumoniae PLY induces TLR2 expression via a TLR4-MyD88-NF-κB-dependent signaling pathway. Interestingly, tumor suppressor CYLD acts as a negative regulator of S. pneumoniae-induced TLR2 up-regulation via negative-crosstalk with NF-κB signaling. Conclusion Our study thus provides novel insights into the regulation of TLR2 expression in mixed bacterial infections.

  6. Regulation of the vapBC-1 toxin-antitoxin locus in nontypeable Haemophilus influenzae.

    Directory of Open Access Journals (Sweden)

    Susan D Cline

    Full Text Available Nontypeable Haemophilus influenzae (NTHi are human-adapted commensal bacteria that can cause a number of chronic mucosal infections, including otitis media and bronchitis. One way for these organisms to survive antibiotic therapy and cause recurrent disease is to stop replicating, as most antimicrobials target essential biosynthetic pathways. Toxin-antitoxin (TA gene pairs have been shown to facilitate entry into a reversible bacteriostatic state. Characteristically, these operons encode a protein toxin and an antitoxin that associate following translation to form a nontoxic complex, which then binds to and regulates the cognate TA promoter. Under stressful conditions, the labile antitoxin is degraded and the complex disintegrates, freeing the stable toxin to facilitate growth arrest. How these events affected the regulation of the TA locus, as well as how the transcription of the operon was subsequently returned to its normal state upon resumption of growth, was not fully understood. Here we show that expression of the NTHi vapBC-1 TA locus is repressed by a complex of VapB-1 and VapC-1 under conditions favorable for growth, and activated by the global transactivator Factor for Inversion Stimulation (Fis upon nutrient upshift from stationary phase. Further, we demonstrate for the first time that the VapC-1 toxin alone can bind to its cognate TA locus control region and that the presence of VapB-1 directs the binding of the VapBC-1 complex in the transcriptional regulation of vapBC-1.

  7. Loss of plasmids containing cloned inserts coding for novobiocin resistance or novobiocin sensitivity in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, J.K.; Spikes, D.; Ledbetter, M.

    1984-06-01

    Plasmids pNov1 and pNov1s, coding for resistance and sensitivity to novobiocin, respectively, were readily lost from wild-type Haemophilus influenzae but retained in a strain lacking an inducible defective prophage. The plasmid loss could be partly or wholly eliminated by a low-copy-number mutation in the plasmid or by the presence of certain antibiotic resistance markers in the host chromosome. Release of both phage HP1c1, measured by plaque assay, and defective phage, measured by electron microscopy, was increased when the plasmids were present. The frequency of recombination between pNov1 and the chromosome, causing the plasmid to be converted to pNov1s, could under some circumstances be decreased from the normal 60 to 70% to below 10% by the presence of a kanamycin resistance marker in the chromosome. This suggested that a gene product coded for by the plasmid, the expression of which was affected by the kanamycin resistance marker, was responsible for the high recombination frequency. Evidence was obtained from in vitro experiments that the gene product was a gyrase.

  8. Single-stranded regions in transforming deoxyribonucleic acid after uptake by competent Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Sedgwick, B.; Setlow, J.K.

    1976-02-01

    About 15% of donor deoxyribonucleic acid (DNA) is single stranded immediately after uptake into competent Haemophilus influenzae wild-type cells, as judged by its sensitivity to S1 endonuclease. This amount decreases to 4 to 5% by 30 min after uptake. Mutants which are defective in the covalent association of recipient and donor DNA form little or no S1 endonuclease-sensitive donor. At 17 C donor DNA taken up by the wild type contains single-stranded regions although there is no observable association, either covalent or noncovalent. The single-stranded regions are at the ends of donor DNA molecules, as judged by the unchanged sedimentation velocity after S1 endonuclease digestion. The amount of single-stranded donor remains constant at 17 C for more than 60 min after uptake, suggesting that the decrease observed at 37 C is the result of association of single-stranded ends with single-stranded regions of recipient cell DNA. Three sequential steps necessary for the integration of donor DNA into recipient DNA are proposed: the synthesis of single-stranded regions in recipient DNA, the interaction of donor DNA with recipient DNA resulting in the production of single-stranded ends on donor DNA, and the stable pairing of homologous single-stranded regions. (auth)

  9. Molecular epidemiology of nontypeable Haemophilus influenzae causing community-acquired pneumonia in adults.

    Directory of Open Access Journals (Sweden)

    Carmen Puig

    Full Text Available Nontypeable Haemophilus influenzae (NTHi is an opportunistic pathogen which causes a variety of respiratory infections. The objectives of the study were to determine its antimicrobial susceptibility, to characterize the β-lactam resistance, and to establish a genetic characterization of NTHi isolates. Ninety-five NTHi isolates were analyzed by pulsed field gel electrophoresis (PFGE and multi locus sequence typing (MLST. Antimicrobial susceptibility was determined by microdilution, and the ftsI gene (encoding penicillin-binding protein 3, PBP3 was PCR amplified and sequenced. Thirty (31.6% isolates were non-susceptible to ampicillin (MIC ≥ 2 mg/L, with 10 of them producing β-lactamase type TEM-1 as a resistance mechanism. After ftsI sequencing, 39 (41.1% isolates showed amino acid substitutions in PBP3, with Asn526 → Lys being the most common (69.2%. Eighty-four patients were successfully treated with amoxicillin/clavulanic acid, ceftriaxone and levofloxacin. Eight patients died due either to aspiration or complication of their comorbidities. In conclusion, NTHi causing CAP in adults shows high genetic diversity and is associated with a high rate of reduced susceptibility to ampicillin due to alterations in PBP3. The analysis of treatment and outcomes demonstrated that NTHi strains with mutations in the ftsI gene could be successfully treated with ceftriaxone or fluoroquinolones.

  10. Acid-soluble breakdown of homologous deoxyribonucleic acid adsorbed by Haemophilus influenzae: its biological significance

    Energy Technology Data Exchange (ETDEWEB)

    Stuy, J.H.

    1974-11-01

    Competent bacteria of Haemophilus influenzae strain Rd were exposed to various kinds of radioactive deoxyribonucleic acid (DNA) for short periods of time and at relatively low temperature. The fate of phage HP1 DNA was studied most extensively. Adsorbed DNA was partially acid solubilized by lysogens and by nonlysogens with very similar kinetics. The biological activity of the DNA decreased extensively in both lysogenic and nonlysogenic recipients. 2,4-Dinitrophenol had no effect on the acid solubilization but largely abolished the biological inactivation. Inactivation kinetics for three different markers and for the triple combination were roughly the same. The presence of 2,4-dinitrophenol in the medium, or the HP1 prophage in the chromosome, did not alter this observation. This suggests that acid solubilization involves the destruction of whole DNA molecules. In view of the absence of DNA homology between phage and host, it is concluded that acid-soluble breakdown of adsorbed transforming DNA is not an integral part of the donor DNA integration process. Behavior of mutant bacteria indicates that neither exonuclease III nor exonuclease V is involved.

  11. Effects of x-irradiation on a temperate bacteriophage of Haemophilus influenzae. [UV radiation

    Energy Technology Data Exchange (ETDEWEB)

    Boling, M.E.; Randolph, M.L.

    1977-04-01

    The inactivation of bacteriophage HPlcl by x rays in a complex medium was found to be exponential, with a D/sub 0/ (the x-ray exposure necessary to reduce the survival of the phage to 37 percent) of approximately 90 kR. Analysis of results of sucrose sedimentation of DNA from x-irradiated whole phage showed that the D/sub 0/ for intactness of single strands was about 105 kR, and for intactness of double strands, it was much higher. The D/sub 0/ for attachment of x-irradiated phage to the host was roughly estimated as about 1,100 kR. Loss of DNA from the phage occurred and was probably due to lysis of the phage by x irradiation, but the significance of the damage is not clear. The production of single-strand breaks approaches the rate of survival loss after x irradiation. However, single-strand breaks produced by uv irradiation, in the presence of H/sub 2/O/sub 2/, equivalent to 215 kR of x rays, showed no lethal effect on the phage. Although uv-sensitive mutants of the host cell, Haemophilus influenzae, have been shown to reactivate uv-irradiated phage less than does the wild-type host cell, x-irradiated phage survive equally well on the mutants as on the wild type, a fact suggesting that other repair systems are involved in x-ray repair.

  12. Structural basis for haem piracy from host haemopexin by Haemophilus influenzae.

    Science.gov (United States)

    Zambolin, Silvia; Clantin, Bernard; Chami, Mohamed; Hoos, Sylviane; Haouz, Ahmed; Villeret, Vincent; Delepelaire, Philippe

    2016-05-18

    Haemophilus influenzae is an obligate human commensal/pathogen that requires haem for survival and can acquire it from several host haemoproteins, including haemopexin. The haem transport system from haem-haemopexin consists of HxuC, a haem receptor, and the two-partner-secretion system HxuB/HxuA. HxuA, which is exposed at the cell surface, is strictly required for haem acquisition from haemopexin. HxuA forms complexes with haem-haemopexin, leading to haem release and its capture by HxuC. The key question is how HxuA liberates haem from haemopexin. Here, we solve crystal structures of HxuA alone, and HxuA in complex with the N-terminal domain of haemopexin. A rational basis for the release of haem from haem-haemopexin is derived from both in vivo and in vitro studies. HxuA acts as a wedge that destabilizes the two-domains structure of haemopexin with a mobile loop on HxuA that favours haem ejection by redirecting key residues in the haem-binding pocket of haemopexin.

  13. Shp2 Deficiency Impairs the Inflammatory Response Against Haemophilus influenzae by Regulating Macrophage Polarization.

    Science.gov (United States)

    Zhao, Lifang; Xia, Jingyan; Li, Tiantian; Zhou, Hui; Ouyang, Wei; Hong, Zhuping; Ke, Yuehai; Qian, Jing; Xu, Feng

    2016-08-15

    Macrophages can polarize and differentiate to regulate initiation, development, and cessation of inflammation during pulmonary infection with nontypeable Haemophilus influenzae (NTHi). However, the underlying molecular mechanisms driving macrophage phenotypic differentiation are largely unclear. Our study investigated the role of Shp2, a Src homology 2 domain-containing phosphatase, in the regulation of pulmonary inflammation and bacterial clearance. Shp2 levels were increased upon NTHi stimulation. Selective inhibition of Shp2 in mice led to an attenuated inflammatory response by skewing macrophages toward alternatively activated macrophage (M2) polarization. Upon pulmonary NTHi infection, Shp2(-/-) mice, in which the gene encoding Shp2 in monocytes/macrophages was deleted, showed an impaired inflammatory response and decreased antibacterial ability, compared with wild-type controls. In vitro data demonstrated that Shp2 regulated activated macrophage (M1) gene expression via activation of p65-nuclear factor-κB signaling, independent of p38 and extracellular regulated kinase-mitogen-activated proteins kinase signaling pathways. Taken together, our study indicates that Shp2 is required to orchestrate macrophage function and regulate host innate immunity against pulmonary bacterial infection.

  14. Antisera Against Certain Conserved Surface-Exposed Peptides of Nontypeable Haemophilus influenzae Are Protective.

    Directory of Open Access Journals (Sweden)

    Paul W Whitby

    Full Text Available Nontypeable Haemophilus influenzae (NTHi cause significant disease, including otitis media in children, exacerbations of chronic obstructive pulmonary disease, and invasive disease in susceptible populations. No vaccine is currently available to prevent NTHi disease. The interactions of NTHi and the human host are primarily mediated by lipooligosaccharide and a complex array of surface-exposed proteins (SEPs that act as receptors, sensors and secretion systems. We hypothesized that certain SEPs are present in all NTHi strains and that a subset of these may be antibody accessible and represent protective epitopes. Initially we used 15 genomic sequences available in the GenBank database along with an additional 11 genomic sequences generated by ourselves to identify the core set of putative SEPs present in all strains. Using bioinformatics, 56 core SEPs were identified. Molecular modeling generated putative structures of the SEPs from which potential surface exposed regions were defined. Synthetic peptides corresponding to ten of these highly conserved surface-exposed regions were used to raise antisera in rats. These antisera were used to assess passive protection in the infant rat model of invasive NTHi infection. Five of the antisera were protective, thus demonstrating their in vivo antibody accessibility. These five peptide regions represent potential targets for peptide vaccine candidates to protect against NTHi infection.

  15. Nontypeable Haemophilus influenzae induces sustained lung oxidative stress and protease expression.

    Directory of Open Access Journals (Sweden)

    Paul T King

    Full Text Available Nontypeable Haemophilus influenzae (NTHi is a prevalent bacterium found in a variety of chronic respiratory diseases. The role of this bacterium in the pathogenesis of lung inflammation is not well defined. In this study we examined the effect of NTHi on two important lung inflammatory processes 1, oxidative stress and 2, protease expression. Bronchoalveolar macrophages were obtained from 121 human subjects, blood neutrophils from 15 subjects, and human-lung fibroblast and epithelial cell lines from 16 subjects. Cells were stimulated with NTHi to measure the effect on reactive oxygen species (ROS production and extracellular trap formation. We also measured the production of the oxidant, 3-nitrotyrosine (3-NT in the lungs of mice infected with this bacterium. NTHi induced widespread production of 3-NT in mouse lungs. This bacterium induced significantly increased ROS production in human fibroblasts, epithelial cells, macrophages and neutrophils; with the highest levels in the phagocytic cells. In human macrophages NTHi caused a sustained, extracellular production of ROS that increased over time. The production of ROS was associated with the formation of macrophage extracellular trap-like structures which co-expressed the protease metalloproteinase-12. The formation of the macrophage extracellular trap-like structures was markedly inhibited by the addition of DNase. In this study we have demonstrated that NTHi induces lung oxidative stress with macrophage extracellular trap formation and associated protease expression. DNase inhibited the formation of extracellular traps.

  16. Nontypable Haemophilus influenzae Displays a Prevalent Surface Structure Molecular Pattern in Clinical Isolates

    Science.gov (United States)

    Mauro, Silvia; Hood, Derek W.; Viadas, Cristina; Calatayud, Laura; Morey, Pau; Servin, Alain; Liñares, Josefina; Oliver, Antonio; Bengoechea, José Antonio; Garmendia, Junkal

    2011-01-01

    Non-typable Haemophilus influenzae (NTHi) is a Gram negative pathogen that causes acute respiratory infections and is associated with the progression of chronic respiratory diseases. Previous studies have established the existence of a remarkable genetic variability among NTHi strains. In this study we show that, in spite of a high level of genetic heterogeneity, NTHi clinical isolates display a prevalent molecular feature, which could confer fitness during infectious processes. A total of 111 non-isogenic NTHi strains from an identical number of patients, isolated in two distinct geographical locations in the same period of time, were used to analyse nine genes encoding bacterial surface molecules, and revealed the existence of one highly prevalent molecular pattern (lgtF+, lic2A+, lic1D+, lic3A+, lic3B+, siaA−, lic2C+, ompP5+, oapA+) displayed by 94.6% of isolates. Such a genetic profile was associated with a higher bacterial resistance to serum mediated killing and enhanced adherence to human respiratory epithelial cells. PMID:21698169

  17. Haemophilus influenzae responds to glucocorticoids used in asthma therapy by modulation of biofilm formation and antibiotic resistance

    Science.gov (United States)

    Earl, Chris S; Keong, Teh Wooi; An, Shi-qi; Murdoch, Sarah; McCarthy, Yvonne; Garmendia, Junkal; Ward, Joseph; Dow, J Maxwell; Yang, Liang; O’Toole, George A; Ryan, Robert P

    2015-01-01

    Glucocorticosteroids are used as a main treatment to reduce airway inflammation in people with asthma who suffer from neutrophilic airway inflammation, a condition frequently associated with Haemophilus influenzae colonization. Here we show that glucocorticosteroids have a direct influence on the behavior of H. influenzae that may account for associated difficulties with therapy. Using a mouse model of infection, we show that corticosteroid treatment promotes H. influenzae persistence. Transcriptomic analysis of bacteria either isolated from infected mouse airway or grown in laboratory medium identified a number of genes encoding regulatory factors whose expression responded to the presence of glucocorticosteroids. Importantly, a number of these corticosteroid-responsive genes also showed elevated expression in H. influenzae within sputum from asthma patients undergoing steroid treatment. Addition of corticosteroid to H. influenzae led to alteration in biofilm formation and enhanced resistance to azithromycin, and promoted azithromycin resistance in an animal model of respiratory infection. Taken together, these data strongly suggest that H. influenzae can respond directly to corticosteroid treatment in the airway potentially influencing biofilm formation, persistence and the efficacy of antibiotic treatment. PMID:25995336

  18. N-Nitrosocarbaryl-induced mutagenesis in Haemophilus influenzae strains deficient in repair and recombination.

    Science.gov (United States)

    Beattie, K L

    1975-02-01

    Mutagenesis was studied in repair- and recombination-deficient strains of Haemophilus influenzae after treatment with N-nitrosocarbaryl (NC). Three different strains of H. influenzae carrying mutations affecting excision-repair of UV-induced pyrimidine dimers exhibited normal repair of premutational lesions (as detected by decreased mutation yield resulting from post-treatment DNA synthesis delay) and normal nonreplicative mutation fixation. This indicated that neither of these phenomena are caused by the smae repair mechanism that removes UV-induced pyrimidine dimers from the DNA. The recombination-deficient mutant recI is apparently deficient in the replication-dependent mode of NC-induced mutation fixation. This conclusion is based on the following results: (I) NC-induced mutagenesis is lower in the recI strain than in rec+ cells. (2) Repair of premutational lesions (which depends on the existence of replication-dependent mutation fixation for its detection) was not detected in the recI strain. (3) When nonreplicative mutation fixation and final mutation frequency were measured in the same experiment, about I/4 to I/3 of the final mutation yield could be accounted for by nonreplicative mutation fixation in the rec+ strain, whereas all of the mutation could be accounted for in the recI strain by the nonreplicative mutation fixation. (4) When mutation fixation in strain dna9 recI was followed at the permissive (36 degrees) and nonpermissive (41 degrees) temperatures, it became apparent that in the recI strain replication-dependent mutation fixation occurs at early times, but these newly fixed mutations are unstable and disappear at later times, leaving only the mutations fixed by the nonreplicative process. The recI strain exhibits normal repair of NC-induced single-strand breaks or alkali-labile bonds in the DNA labeled before treatment, but is slow in joining discontinuties present in DNA synthesized after treatment. The results are consistent with the idea that

  19. The capsule biosynthesis locus of Haemophilus influenzae show conspicuous similarity to the corresponding locus in Haemophilus sputorum and may have been recruited from this species by horizontal gene transfer

    DEFF Research Database (Denmark)

    Nielsen, Signe Maria; de Gier, Camilla; Dimopoulou, Chrysoula;

    2015-01-01

    in export and processing of the capsular material, show high similarity to the corresponding genes in capsulate lineages of the pathogenic species Haemophilus influenzae; indeed, standard bexA and bexB PCRs for detection of capsulated strains of H. influenzae give positive results with strains of H......The newly described species Haemophilus sputorum has been cultured from the upper respiratory tract of humans and appears to have little pathogenic potential. The species encode a capsular biosynthesis locus of approximately 12 kb composed of three distinct regions. Region I and III genes, involved....... sputorum. Three open reading frames are present in region II of the sequenced strain of H. sputorum, of which a putative phosphotransferase showed homology with corresponding genes from H. influenzae serotype c and f. Phylogenetic analysis of housekeeping genes from 24 Pasteurellaceae species showed that H...

  20. Role of Sialic Acid and Complex Carbohydrate Biosynthesis in Biofilm Formation by Nontypeable Haemophilus influenzae in the Chinchilla Middle Ear

    OpenAIRE

    Jurcisek, Joseph; Greiner, Laura; Watanabe, Hiroshi; Zaleski, Anthony; Apicella, Michael A.; Bakaletz, Lauren O.

    2005-01-01

    Nontypeable Haemophilus influenzae (NTHI) is an important pathogen in respiratory tract infections, including otitis media (OM). NTHI forms biofilms in vitro as well as in the chinchilla middle ear, suggesting that biofilm formation in vivo might play an important role in the pathogenesis and chronicity of OM. We've previously shown that SiaA, SiaB, and WecA are involved in biofilm production by NTHI in vitro. To investigate whether these gene products were also involved in biofilm production...

  1. In vitro and in vivo synergism between amoxicillin and clavulanic acid against ampicillin-resistant Haemophilus influenzae type b.

    OpenAIRE

    Yogev, R; Melick, C; Kabat, W J

    1981-01-01

    Eight strans of ampicillin-resistant beta-lactamase-producing Haemophilus influenzae type b were studied in vitro for synergy between amoxicillin and clavulanic acid. The minimal inhibitory concentrations for amoxicillin alone were 6.25 to 12.5 microgram/ml, and for clavulanic acid alone they were 12.5 to 25 microgram/ml. However, seven of eight strains were inhibited by a combination of 0.36 microgram of amoxicillin and 0.36 microgram of clavulanic acid per ml. Infant rat models of bacteremi...

  2. Tests for the mutagenic action of a number of chemicals on Haemophilus influenzae with special emphasis on hydrazine

    Energy Technology Data Exchange (ETDEWEB)

    Kimball, R.F.; Hirsch, B.F.

    1975-01-01

    A number of chemicals have been tested for their ability to produce novobiocin-resistant mutants in Haemophilus influenzae. Of these, hydrazine (HZ) proved unique because it induced a fairly high incidence of mutation without killing significant numbers of cells at concentrations ranging over nearly four orders of magnitude. Moreover, its dose--effect curve increased very slowly initially and reached a relatively low maximum. It is suggested that HZ may be acting as both a mutagen and an antimutagen in this system. (auth)

  3. Genetic control of prophage induction in Haemophilus influenzae after exposure to psoralen plus near-uv light

    Energy Technology Data Exchange (ETDEWEB)

    George, M.; Notani, N.K.

    1980-09-01

    Prophage S2 could be induced by psoralen plus near-uv light (PNUV) from a wild-type strain of Haemophilus influenzae, from uv light-sensitive strains uvr-1 and uvr-2, and PNUV-sensitive strains PSO1 and PSO7, but not from a recombination-deficient strain, rec-1. The levels of prophage induction were comparable in the wild type and an ATP-dependent DNase-deficient strain, KW31, even though the PNUV-induced degradation in the latter strain was considerably lower. Prophage induction could be observed even with chloramphenicol present before, during, and 30 min after PNUV treatment.

  4. Meningitis Due to Ampicillin-and Chloramphenicol-Resistant Haemophilus influenzae Type B in Canada. Case Report and Review

    Directory of Open Access Journals (Sweden)

    Amin Kabani

    1990-01-01

    Full Text Available The first report of a case of ampicillin- and chloramphenicol-resistant Haemophilus influenzae type b invasive infection in Canada is described in a four-month-old male with meningitis. He was treated with cefotaxime 200 mg/kg/day divided every 6 h and dexamethasone 0.6 mg/kg/day divided every 6 h, eventually recovering after a complicated course. Follow-up at 21 months showed mild to moderate global developmental delay. While chloramphenicol resistance is rare in North America, a case of meningitis initially unresponsive to ampicillin and chloramphenicol must be considered suspect for resistance. Third generation cephalosporins should be used for resistant cases.

  5. Repair and actio spectrum of oxygen-independent lethality of near uv light on Haemophilus influenzae and lack of mutation

    Energy Technology Data Exchange (ETDEWEB)

    Cabrera-Juarez, E.; Setlow, J.K.

    1980-01-01

    Haemophilus influenzae has been inactivated anaerobically at 313, 334, 365, and 405 nm, and exhibits the greatest sensitivity at 334 nm. The rec1 and uvr1 mutants show the greatest increase in sensitivity over the wild-type at 313 nm, but differences could be seen also at the other wavelengths. Anaerobic irradiation is less effective for killing at all the wavelengths than irradiation under aerobic conditions, but the greatest difference was observed at 365 nm. No induced mutation was seen as a result of anaerobic irradiation at 334 nm, although purified transforming DNA can be mutated at this wavelength.

  6. Oropharyngeal colonization by Haemophilus influenzae in healthy children from Taubaté (São Paulo, prior to the Haemophilus influenzae type b vaccination program in Brazil Colonização da orofaringe de crianças saudáveis de Taubaté (São Paulo por Haemophilus influenzae, antes da introdução da vacina contra Haemophilus influenzae do tipo b no Brasil

    Directory of Open Access Journals (Sweden)

    Lucia Ferro Bricks

    2004-01-01

    Full Text Available Haemophilus influenzae is one of the most important bacterial agents of otitis and sinusitis. H. influenzae type b (Hib is one of the main causes of meningitis, pneumonia, and septicemia in nonvaccinated children under 6 years of age. The aims of this study were to determine the prevalence of H. influenzae and Hib oropharyngeal colonization prior to the onset of the Hib vaccination program in Brazil in previously healthy children and to assess the susceptibility profile of this microorganism to a selected group of antimicrobials that are used to treat acute respiratory infections. METHOD: Cultures of Haemophilus influenzae were made from oropharynx swabs from 987 children under 6 years of age who were enrolled in 29 day-care centers in Taubaté (a city of São Paulo state, Brazil between July and December 1998. RESULTS: The prevalence of H. influenzae carriers was 17.4%, and only 5.5% of the strains were beta-lactamase producers. The prevalence of Hib carriers was high, 7.3% on average (range, 0.0 - 33.3%. CONCLUSIONS: The low prevalence of colonization by penicillin-resistant strains indicates that it is not necessary to substitute ampicilin or amoxicilin to effectively treat otitis and sinusitis caused by H. influenzae in Taubaté.Haemophilus influenzae é um dos mais importantes agentes bacterianos de otites e sinusites. Em crianças menores de seis anos de idade não vacinadas contra o H. influenzae do tipo b (Hib, essa bactéria é uma das principais causadoras de meningite, pneumonia e sepse. O objetivo deste estudo foi determinar a prevalência da colonização da orofaringe de crianças previamente saudáveis por H. influenzae e Hib e avaliar o perfil de suscetibilidade desses microorganismos a um grupo seleto de antimicrobianos, que habitualmente são utilizados para tratar as infecções respiratórias agudas. MÉTODO: Foram colhidos swabs da orofaringe de 987 crianças menores de seis anos de idade que freqüentavam 29 creches da

  7. [Activity of cefpodoxime and other oral beta-lactams against Haemophilus influenzae and Streptococcus pneumoniae with different susceptibilities to penicillin].

    Science.gov (United States)

    Fenoll, A; Robledo, O; Lerma, M; Giménez, M J; Cebrián, L; Casal, J; Aguilar, L; Gómez-Lus, M L

    2006-03-01

    This study explores the influence on the intrinsic activity of different oral beta-lactams of beta-lactamase production in Haemophilus influenzae and penicillin resistance in Streptococcus pneumoniae. Three substudies were performed: a) a general susceptibility study, analyzing 550 strains received by the Spanish Laboratorio de Referencia de Neumococos throughout February and March 2005; b) a study on the influence of penicillin resistance on the activity of beta-lactams, analyzing 251 penicillin-susceptible strains (MICor=2 mg/l) randomly chosen among those received by the Spanish Laboratorio de Referencia de Neumococos throughout 2005; and c) an H. influenzae susceptibility study analyzing 150 strains received by Instituto Valenciano de Microbiologia throughout 2005. A total of 71% of S. pneumoniae strains were susceptible to penicillin, 21% exhibited intermediate resistance and 8% strains presented full resistance. H. influenzae beta-lactamase production rate was 18.6%. Of the non-beta-lactamase-producing strains, 3% were not susceptible to ampicillin. Cefpodoxime and cefixime exhibited the highest intrinsic activity against H. influenzae, while amoxicillin and cefpodoxime were the most active compounds against S. pneumoniae. All H. influenzae strains were susceptible to oral cephalosporins and amoxicillin/clavulanic acid. The increase in penicillin resistance in S. pneumoniae influenced cefixime, cefaclor and cefuroxime to a higher degree than amoxicillin and cefpodoxime.

  8. Emergence of Non-Serotype b Encapsulated Haemophilus influenzae as a Cause of Pediatric Meningitis in Northwestern Ontario

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    Pouya Sadeghi-Aval

    2013-01-01

    Full Text Available Before the introduction of the conjugate vaccine, Haemophilus influenzae serotype b (Hib was the leading cause of bacterial meningitis in children. Although successful in reducing Hib cases, the vaccine confers no protection against other serotypes of H influenzae, such as a (Hia, or f (Hif. The emergence of invasive disease caused by non-Hib in northwestern Ontario (38 cases between 2002 and 2008 with predominance of Hia was previously reported by the authors. At that time, no cases of pediatric meningitis caused by H influenzae were recorded in the region. Continued surveillance identified 12 new cases of invasive non-Hib between January 2009 and July 2011. Among these cases, three young children developed meningitis with severe complications caused by Hia or Hif. The present article describes these cases along with the characteristics of recent H influenzae isolates from the region, (ie, their genetic background and antibiotic sensitivity. The findings point to the clonal nature of circulating Hia strains as well as to an increase in frequency and severity of pediatric invasive H influenzae infections in northwestern Ontario.

  9. Investigations into genome diversity of Haemophilus influenzae using whole genome sequencing of clinical isolates and laboratory transformants

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    Power Peter M

    2012-11-01

    Full Text Available Abstract Background Haemophilus influenzae is an important human commensal pathogen associated with significant levels of disease. High-throughput DNA sequencing was used to investigate differences in genome content within this species. Results Genomic DNA sequence was obtained from 85 strains of H. influenzae and from other related species, selected based on geographical site of isolation, disease association and documented genotypic and phenotypic differences. When compared by Mauve alignment these indicated groupings of H. influenzae that were consistent with previously published analyses; capsule expressing strains fell into two distinct groups and those of serotype b (Hib were found in two closely positioned lineages. For 18 Hib strains representing both lineages we found many discrete regions (up to 40% of the total genome displaying sequence variation when compared to a common reference strain. Evidence that this naturally occurring pattern of inter-strain variation in H. influenzae can be mediated by transformation was obtained through sequencing DNA obtained from a pool of 200 independent transformants of a recipient (strain Rd using donor DNA from a heterologous Hib strain (Eagan. Conclusion Much of the inter-strain variation in genome sequence in H. influenzae is likely the result of inter-strain exchanges of DNA, most plausibly through transformation.

  10. Impacto da vacinação contra o Haemophilus influenzae b na redução de meningites, Goiás

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    Simões Luciana Leite Pineli

    2004-01-01

    Full Text Available OBJETIVO: Avaliar o impacto da vacinação contra o Haemophilus influenzae b na incidência de meningites em crianças menores de cinco anos de idade. MÉTODOS: Utilizou-se o delineamento tipo "antes-depois" para comparar as taxas de incidência de meningites por Haemophilus influenzae b nos períodos pré-vacinação (julho/95-junho/99 e pós-vacinação (julho/99-junho/2001 no Estado de Goiás. A definição de caso de meningite bacteriana seguiu os critérios da Organização Mundial de Saúde. As taxas de meningite por Streptococcus pneumoniae e Neisseria. meningitidis foram utilizadas para efeito de comparação. Para análise estatística foram utilizados o teste de chi2 e o t de Student. Valores de p<0,05 foram considerados estatisticamente significantes. RESULTADOS: Foi detectada meningite bacteriana aguda em 979 crianças no período de estudo. A incidência de meningite por Haemophilus influenzae b diminuiu de 10,8x10(5 no período pré-vacinal para 2,3x10(5 no segundo ano pós-vacina, significando 78% de redução no risco, principalmente na faixa etária de 7-23 meses (p<0,05. Foram prevenidos 65 casos de meningite por Haemophilus influenzae b. Observou-se aumento na incidência de meningite por S. pneumoniae. Foi observada falha vacinal em um caso. CONCLUSÕES: Expressivo declínio da incidência de meningite por Haemophilus influenzae b foi detectado, precocemente, logo após o primeiro ano de introdução da vacina contra o Haemophilus influenzae b. Assim, se faz necessária a vigilância contínua com instrumental de alta acurácia para: (i detectar re-emergência do Haemophilus influenzae b; (ii avaliar possibilidade de falha vacinal; (iii identificar mudanças no padrão dos sorotipos do H. influenzae.

  11. Amoxicillin treatment of experimental acute otitis media caused by Haemophilus influenzae with non-beta-lactamase-mediated resistance to beta-lactams: aspects of virulence and treatment.

    OpenAIRE

    Melhus, A; Janson, H; Westman, E.; Hermansson, A.; Forsgren, A; Prellner, K

    1997-01-01

    Through alterations primarily in the penicillin-binding proteins, a non-beta-lactamase-mediated resistance to beta-lactams has evolved in Haemophilus influenzae. The virulence of these chromosomally changed strains has been questioned. To ascertain whether these alterations involve a reduction in virulence of H. influenzae and whether they could be advantageous for the bacterium during amoxicillin treatment of acute otitis media, a total of 70 Sprague-Dawley rats were challenged with a suscep...

  12. Preclinical evaluation of a Haemophilus influenzae type b conjugate vaccine process intended for technology transfer.

    Science.gov (United States)

    Hamidi, Ahd; Verdijk, Pauline; Kreeftenberg, Hans

    2014-01-01

    Introduction of Haemophilus influenzae type b (Hib) vaccine in low- and middle-income countries has been limited by cost and availability of Hib conjugate vaccines for a long time. It was previously recognized by the Institute for Translational Vaccinology (Intravacc, originating from the former Vaccinology Unit of the National Institute of Public Health [RIVM] and the Netherlands Vaccine Institute [NVI]) that local production of a Hib conjugate vaccine would increase the affordability and sustainability of the vaccine and thereby help to speed up Hib introduction in these countries. A new affordable and a non-infringing production process for a Hib conjugate vaccine was developed, including relevant quality control tests, and the technology was transferred to a number of vaccine manufacturers in India, Indonesia, and China. As part of the Hib technology transfer project managed by Intravacc, a preclinical toxicity study was conducted in the Netherlands to test the safety and immunogenicity of this new Hib conjugate vaccine. The data generated by this study were used by the technology transfer partners to accelerate the clinical development of the new Hib conjugate vaccine. A repeated dose toxicity and local tolerance study in rats was performed to assess the reactogenicity and immunogenicity of a new Hib conjugate vaccine compared to a licensed vaccine. The results showed that the vaccine was well tolerated and immunogenic in rats, no major differences in both safety and immunogenicity in rats were found between the vaccine produced according to the production process developed by Intravacc and the licensed one. Rats may be useful to verify the immunogenicity of Hib conjugate vaccines and for preclinical evaluation. In general, nonclinical evaluation of the new Hib conjugate vaccine, including this proof of concept (safety and immunogenicity study in rats), made it possible for technology transfer partners, having implemented the original process with no changes

  13. HPAEC-PAD quantification of Haemophilus influenzae type b polysaccharide in upstream and downstream samples.

    Science.gov (United States)

    van der Put, Robert M F; de Haan, Alex; van den IJssel, Jan G M; Hamidi, Ahd; Beurret, Michel

    2015-11-27

    Due to the rapidly increasing introduction of Haemophilus influenzae type b (Hib) and other conjugate vaccines worldwide during the last decade, reliable and robust analytical methods are needed for the quantitative monitoring of intermediate samples generated during fermentation (upstream processing, USP) and purification (downstream processing, DSP) of polysaccharide vaccine components. This study describes the quantitative characterization of in-process control (IPC) samples generated during the fermentation and purification of the capsular polysaccharide (CPS), polyribosyl-ribitol-phosphate (PRP), derived from Hib. Reliable quantitative methods are necessary for all stages of production; otherwise accurate process monitoring and validation is not possible. Prior to the availability of high performance anion exchange chromatography methods, this polysaccharide was predominantly quantified either with immunochemical methods, or with the colorimetric orcinol method, which shows interference from fermentation medium components and reagents used during purification. Next to an improved high performance anion exchange chromatography-pulsed amperometric detection (HPAEC-PAD) method, using a modified gradient elution, both the orcinol assay and high performance size exclusion chromatography (HPSEC) analyses were evaluated. For DSP samples, it was found that the correlation between the results obtained by HPAEC-PAD specific quantification of the PRP monomeric repeat unit released by alkaline hydrolysis, and those from the orcinol method was high (R(2)=0.8762), and that it was lower between HPAEC-PAD and HPSEC results. Additionally, HPSEC analysis of USP samples yielded surprisingly comparable results to those obtained by HPAEC-PAD. In the early part of the fermentation, medium components interfered with the different types of analysis, but quantitative HPSEC data could still be obtained, although lacking the specificity of the HPAEC-PAD method. Thus, the HPAEC

  14. Placental transfer of Haemophilus influenzae type b antibodies in malnourished pregnant women

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    Rejane S. Cavalcante

    2008-02-01

    Full Text Available This study evaluated the vaccination response to Haemophilus influenzae type b (Hib in malnourished pregnant women (MN, cord blood (CB and in infants at two and six months of age for comparison with a control group (C. Twenty-eight malnourished pregnant women and 29 pregnant controls were immunized with conjugated Act-HIB® in the third trimester of pregnancy. Blood samples were collected from all before the immunization, during labor (post immunization, and from CB. All infants were immunized with Hib vaccine according to normal vaccine schedule and sera were collected at two and six months of age. Antibody levels to polyribosylribitol phosphate (PRP were similar for both groups. Preimmunization: MN 1.94 µg/mL, C 1.68 µg/mL; post-vaccination: MN 18.53 µg/mL and C 17.55 µg/mL; in CB from MN 14.46 µg/mL and from C 17.04 µg/mL. Infants from MN and C mothers presented respectively at two months: 5.18 µg/mL and 8.60 µg/mL and at six months: MN 3.42 µg/mL and C 2.18 µg/mL. Antibody levels were similar in both groups studied (p = 0.485, however the vertical transmission rate was 14% lower in the MN pregnant group. Levels of antibodies > 0.15 µg/mL were found in all newborns from the MN pregnant group. Pregnant MN presented an immunological response to Hib vaccine similar to group C, however, vertical transmission rate of antibodies to PRP in the MN pregnant group was 14% lower than that in C, suggesting a less efficient passage of antibodies within this group.

  15. Transformed Recombinant Enrichment Profiling Rapidly Identifies HMW1 as an Intracellular Invasion Locus in Haemophilus influenza

    Science.gov (United States)

    Moleres, Javier; Sinha, Sunita; Fernández-Calvet, Ariadna; Porsch, Eric A.; St. Geme, Joseph W.; Nislow, Corey; Redfield, Rosemary J.; Garmendia, Junkal

    2016-01-01

    Many bacterial species actively take up and recombine homologous DNA into their genomes, called natural competence, a trait that offers a means to identify the genetic basis of naturally occurring phenotypic variation. Here, we describe “transformed recombinant enrichment profiling” (TREP), in which natural transformation is used to generate complex pools of recombinants, phenotypic selection is used to enrich for specific recombinants, and deep sequencing is used to survey for the genetic variation responsible. We applied TREP to investigate the genetic architecture of intracellular invasion by the human pathogen Haemophilus influenzae, a trait implicated in persistence during chronic infection. TREP identified the HMW1 adhesin as a crucial factor. Natural transformation of the hmw1 operon from a clinical isolate (86-028NP) into a laboratory isolate that lacks it (Rd KW20) resulted in ~1,000-fold increased invasion into airway epithelial cells. When a distinct recipient (Hi375, already possessing hmw1 and its paralog hmw2) was transformed by the same donor, allelic replacement of hmw2AHi375 by hmw1A86-028NP resulted in a ~100-fold increased intracellular invasion rate. The specific role of hmw1A86-028NP was confirmed by mutant and western blot analyses. Bacterial self-aggregation and adherence to airway cells were also increased in recombinants, suggesting that the high invasiveness induced by hmw1A86-028NP might be a consequence of these phenotypes. However, immunofluorescence results found that intracellular hmw1A86-028NP bacteria likely invaded as groups, instead of as individual bacterial cells, indicating an emergent invasion-specific consequence of hmw1A-mediated self-aggregation. PMID:27124727

  16. Early-Life Intranasal Colonization with Nontypeable Haemophilus influenzae Exacerbates Juvenile Airway Disease in Mice.

    Science.gov (United States)

    McCann, Jessica R; Mason, Stanley N; Auten, Richard L; St Geme, Joseph W; Seed, Patrick C

    2016-07-01

    Accumulating evidence suggests a connection between asthma development and colonization with nontypeable Haemophilus influenzae (NTHi). Specifically, nasopharyngeal colonization of human infants with NTHi within 4 weeks of birth is associated with an increased risk of asthma development later in childhood. Monocytes derived from these infants have aberrant inflammatory responses to common upper respiratory bacterial antigens compared to those of cells derived from infants who were not colonized and do not go on to develop asthma symptoms in childhood. In this study, we hypothesized that early-life colonization with NTHi promotes immune system reprogramming and the development of atypical inflammatory responses. To address this hypothesis in a highly controlled model, we tested whether colonization of mice with NTHi on day of life 3 induced or exacerbated juvenile airway disease using an ovalbumin (OVA) allergy model of asthma. We found that animals that were colonized on day of life 3 and subjected to induction of allergy had exacerbated airway disease as juveniles, in which exacerbated airway disease was defined as increased cellular infiltration into the lung, increased amounts of inflammatory cytokines interleukin-5 (IL-5) and IL-13 in lung lavage fluid, decreased regulatory T cell-associated FOXP3 gene expression, and increased mucus production. We also found that colonization with NTHi amplified airway resistance in response to increasing doses of a bronchoconstrictor following OVA immunization and challenge. Together, the murine model provides evidence for early-life immune programming that precedes the development of juvenile airway disease and corroborates observations that have been made in human children. PMID:27113355

  17. Preparation and testing of a Haemophilus influenzae Type b/Hepatitis B surface antigen conjugate vaccine.

    Science.gov (United States)

    An, So Jung; Woo, Joo Sung; Chae, Myung Hwa; Kothari, Sudeep; Carbis, Rodney

    2015-03-24

    The majority of conjugate vaccines focus on inducing an antibody response to the polysaccharide antigen and the carrier protein is present primarily to induce a T-cell dependent response. In this study conjugates consisting of poly(ribosylribitolphosphate) (PRP) purified from Haemophilus influenzae Type b bound to Hepatitis B virus surface antigen (HBsAg) virus like particles were prepared with the aim of inducing an antibody response to not only the PRP but also the HBsAg. A conjugate consisting of PRP bound to HBsAg via an adipic acid dihydrazide (ADH) spacer induced strong IgG antibodies to both the PRP and HBsAg. When conjugation was performed without the ADH spacer the induction of an anti-PRP response was equivalent to that seen by conjugate with the ADH spacer, however, a negligible anti-HBsAg response was induced. For comparison, PRP was conjugated to diphtheria toxoid (DT) and Vi polysaccharide purified from Salmonella Typhi conjugated to HBsAg both using an ADH spacer. The PRPAH-DT conjugate induced strong anti-PRP and anti-DT responses, the Vi-AHHBsAg conjugate induced a good anti-HBsAg response but not as strong as that induced by the PRPAH-HBsAg conjugate. This study demonstrated that in mice it was possible to induce robust antibody responses to both polysaccharide and carrier protein provided the conjugate has certain physico-chemical properties. A PRPAH-HBsAg conjugate with the capacity to induce anti-PRP and anti-HBsAg responses could be incorporated into a multivalent pediatric vaccine and simplify formulation of such a vaccine.

  18. Resolvin D1 Dampens Pulmonary Inflammation and Promotes Clearance of Nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Croasdell, Amanda; Lacy, Shannon H; Thatcher, Thomas H; Sime, Patricia J; Phipps, Richard P

    2016-03-15

    Nontypeable Haemophilus influenzae (NTHi) is a Gram-negative, opportunistic pathogen that frequently causes ear infections, bronchitis, pneumonia, and exacerbations in patients with underlying inflammatory diseases, such as chronic obstructive pulmonary disease. In mice, NTHi is rapidly cleared, but a strong inflammatory response persists, underscoring the concept that NTHi induces dysregulation of normal inflammatory responses and causes a failure to resolve. Lipid-derived specialized proresolving mediators (SPMs) play a critical role in the active resolution of inflammation by both suppressing proinflammatory actions and promoting resolution pathways. Importantly, SPMs lack the immunosuppressive properties of classical anti-inflammatory therapies. On the basis of these characteristics, we hypothesized that aspirin-triggered resolvin D1 (AT-RvD1) would dampen NTHi-induced inflammation while still enhancing bacterial clearance. C57BL/6 mice were treated with AT-RvD1 and infected with live NTHi. AT-RvD1-treated mice had lower total cell counts and neutrophils in bronchoalveolar lavage fluid, and had earlier influx of macrophages. In addition, AT-RvD1-treated mice showed changes in temporal regulation of inflammatory cytokines and enzymes, with decreased KC at 6 h and decreased IL-6, TNF-α, and cyclooxygenase-2 expression at 24 h post infection. Despite reduced inflammation, AT-RvD1-treated mice had reduced NTHi bacterial load, mediated by enhanced clearance by macrophages and a skewing toward an M2 phenotype. Finally, AT-RvD1 protected NTHi-infected mice from weight loss, hypothermia, hypoxemia, and respiratory compromise. This research highlights the beneficial role of SPMs in pulmonary bacterial infections and provides the groundwork for further investigation into SPMs as alternatives to immunosuppressive therapies like steroids.

  19. Recognition of Nucleoside Monophosphate Substrates by Haemophilus influenzae Class C Acid Phosphatase

    Energy Technology Data Exchange (ETDEWEB)

    Singh, Harkewal; Schuermann, Jonathan P.; Reilly, Thomas J.; Calcutt, Michael J.; Tanner, John J. (Cornell); (UMC)

    2010-12-08

    The e (P4) phosphatase from Haemophilus influenzae functions in a vestigial NAD{sup +} utilization pathway by dephosphorylating nicotinamide mononucleotide to nicotinamide riboside. P4 is also the prototype of class C acid phosphatases (CCAPs), which are nonspecific 5{prime},3{prime}-nucleotidases localized to the bacterial outer membrane. To understand substrate recognition by P4 and other class C phosphatases, we have determined the crystal structures of a substrate-trapping mutant P4 enzyme complexed with nicotinamide mononucleotide, 5{prime}-AMP, 3{prime}-AMP, and 2{prime}-AMP. The structures reveal an anchor-shaped substrate-binding cavity comprising a conserved hydrophobic box that clamps the nucleotide base, a buried phosphoryl binding site, and three solvent-filled pockets that contact the ribose and the hydrogen-bonding edge of the base. The span between the hydrophobic box and the phosphoryl site is optimal for recognizing nucleoside monophosphates, explaining the general preference for this class of substrate. The base makes no hydrogen bonds with the enzyme, consistent with an observed lack of base specificity. Two solvent-filled pockets flanking the ribose are key to the dual recognition of 5{prime}-nucleotides and 3{prime}-nucleotides. These pockets minimize the enzyme's direct interactions with the ribose and provide sufficient space to accommodate 5{prime} substrates in an anti conformation and 3{prime} substrates in a syn conformation. Finally, the structures suggest that class B acid phosphatases and CCAPs share a common strategy for nucleotide recognition.

  20. Identification and Characterization of msf, a Novel Virulence Factor in Haemophilus influenzae.

    Science.gov (United States)

    Kress-Bennett, Jennifer M; Hiller, N Luisa; Eutsey, Rory A; Powell, Evan; Longwell, Mark J; Hillman, Todd; Blackwell, Tenisha; Byers, Barbara; Mell, Joshua C; Post, J Christopher; Hu, Fen Z; Ehrlich, Garth D; Janto, Benjamin A

    2016-01-01

    Haemophilus influenzae is an opportunistic pathogen. The emergence of virulent, non-typeable strains (NTHi) emphasizes the importance of developing new interventional targets. We screened the NTHi supragenome for genes encoding surface-exposed proteins suggestive of immune evasion, identifying a large family containing Sel1-like repeats (SLRs). Clustering identified ten SLR-containing gene subfamilies, each with various numbers of SLRs per gene. Individual strains also had varying numbers of SLR-containing genes from one or more of the subfamilies. Statistical genetic analyses of gene possession among 210 NTHi strains typed as either disease or carriage found a significant association between possession of the SlrVA subfamily (which we have termed, macrophage survival factor, msf) and the disease isolates. The PittII strain contains four chromosomally contiguous msf genes. Deleting all four of these genes (msfA1-4) (KO) resulted in a highly significant decrease in phagocytosis and survival in macrophages; which was fully complemented by a single copy of the msfA1 gene. Using the chinchilla model of otitis media and invasive disease, the KO strain displayed a significant decrease in fitness compared to the WT in co-infections; and in single infections, the KO lost its ability to invade the brain. The singly complemented strain showed only a partial ability to compete with the WT suggesting gene dosage is important in vivo. The transcriptional profiles of the KO and WT in planktonic growth were compared using the NTHi supragenome array, which revealed highly significant changes in the expression of operons involved in virulence and anaerobiosis. These findings demonstrate that the msfA1-4 genes are virulence factors for phagocytosis, persistence, and trafficking to non-mucosal sites.

  1. Acute Exacerbations in COPD and their Control with Oral Immunisation with nontypeable Haemophilus influenzae

    Directory of Open Access Journals (Sweden)

    Robert eClancy

    2011-03-01

    Full Text Available Chronic obstructive pulmonary disease (COPD a term based on the demonstration of irreversible airways obstruction, introduced to unify a range of chronic progressive diseases of the airways consequent upon inhalation of toxins. While disease is initiated and progressed by inhaled toxins, an additional pathway of damage has emerged, with particular relevance to acute exacerbations. Exacerbations of disease due to an increase in the level of intra-bronchial inflammation have taken on a new significance as their role in determining both acute and chronic outcomes is better understood. This ‘second pathway’ of disease is a consequence of bacterial colonisation of damaged airways. Although bacteria have been linked to acute episodes in COPD over 50 years, only recently has quality data on antibiotic usage and the detection of ‘exacerbation isolates’ of non-typeable Haemophilus influenzae (NTHi provided strong argument in support of a pathogenic role. Yet a poor correlation between detection of colonising bacteria and clinical status remained a concern in attempts to explain a role for bacteria in a classical infection model. This presentation discusses a hypothesis that acute exacerbations reflect a T-cell dependent hypersensitivity response to colonising bacteria, with IL-17 dependent accumulation of neutrophils within the bronchus, as the main outcome measure. Critical protection against exacerbations following oral administration of NTHi, an immunotherapy that drives a TH17 Tcell response from Peyer’s patches, reduces the load of intrabronchial bacteria while preventing access of inhaled bacteria into small airways. Immunotherapy augments a physiological ‘loop’ based on aspiration of bronchus content into the gut. A second ‘hypersensitivity’ mechanism may cause bronchospasm – in both COPD and treatment-resistant asthma – due to specific IgE antibody directed against colonising bacteria, as oral NTHi abrogates wheeze in

  2. Early-Life Intranasal Colonization with Nontypeable Haemophilus influenzae Exacerbates Juvenile Airway Disease in Mice.

    Science.gov (United States)

    McCann, Jessica R; Mason, Stanley N; Auten, Richard L; St Geme, Joseph W; Seed, Patrick C

    2016-07-01

    Accumulating evidence suggests a connection between asthma development and colonization with nontypeable Haemophilus influenzae (NTHi). Specifically, nasopharyngeal colonization of human infants with NTHi within 4 weeks of birth is associated with an increased risk of asthma development later in childhood. Monocytes derived from these infants have aberrant inflammatory responses to common upper respiratory bacterial antigens compared to those of cells derived from infants who were not colonized and do not go on to develop asthma symptoms in childhood. In this study, we hypothesized that early-life colonization with NTHi promotes immune system reprogramming and the development of atypical inflammatory responses. To address this hypothesis in a highly controlled model, we tested whether colonization of mice with NTHi on day of life 3 induced or exacerbated juvenile airway disease using an ovalbumin (OVA) allergy model of asthma. We found that animals that were colonized on day of life 3 and subjected to induction of allergy had exacerbated airway disease as juveniles, in which exacerbated airway disease was defined as increased cellular infiltration into the lung, increased amounts of inflammatory cytokines interleukin-5 (IL-5) and IL-13 in lung lavage fluid, decreased regulatory T cell-associated FOXP3 gene expression, and increased mucus production. We also found that colonization with NTHi amplified airway resistance in response to increasing doses of a bronchoconstrictor following OVA immunization and challenge. Together, the murine model provides evidence for early-life immune programming that precedes the development of juvenile airway disease and corroborates observations that have been made in human children.

  3. Effect of epithelial cell type on in vitro invasion of non-typeable Haemophilus influenzae.

    Science.gov (United States)

    Singh, Neeraj Kumar; Kunde, Dale A; Tristram, Stephen G

    2016-10-01

    Non-typeable Haemophilus influenzae (NTHi) have been shown to have variable ability for in vitro invasion with a range of epithelial cells, and increased invasion of BEAS-2B cells has been associated with altered penicillin binding protein3 (PBP3), which is concerning as these strains are increasing worldwide. The aim of the study was to investigate the effect of respiratory cell type and the presence of altered PBP3 on the in vitro invasion of NTHi. A collection of 16 clinical NTHi isolates was established, 7 had normal PBP3, and 9 had altered PBP3 as defined by an N526K substitution. The isolates were tested for invasion of BEAS-2B, NHBE, A549 and NCI-H292 respiratory epithelial cells in vitro using a gentamicin survival assay, with invasion measured as the percentage of intracellular organisms relative to the initial inoculum. The overall median invasion for the 16 NTHi isolates for cell types BEAS-2B, NHBE, A549 and NCI-H292 cells were 3.17, 2.31, 0.11 and 1.52 respectively. The differences were statistically significant for BEAS-2B compared to A549 (P=0.015) and A549 compared to NCI-H292 (P=0.015), and there were also very marked differences in invasion for some individual isolates depending on the cell type used. There was a consistent bias for invasion of isolates with normal versus abnormal PBP3: and this was statistically significant for BEAS-2B (0.07 to 9.90, P=0.031) and A549 cells (0.02 to 1.68, P=0.037). These results show that NTHi invasion of respiratory epithelial cells in vitro is both strain dependant and influenced significantly by the cell line used, and that the association between altered PBP3 and increased invasion is conserved across multiple cell lines.

  4. Prokaryotic High-Level Expression System in Producing Adhesin Recombinant Protein E of Nontypeable Haemophilus influenzae

    Science.gov (United States)

    Tavakoli, Minoo; Bouzari, Saeed; Siadat, Seyed Davar; Najar Peerayeh, Shahin; Jafari, Anis

    2015-01-01

    Background: Adhesion protein E (PE) of Haemophilus influenzae is a 16 - 18 kDa protein with 160 amino acids which causes adhesion to epithelial cells and acts as a major factor in pathogenesis. Objectives: In this study, we performed cloning, expression and purification of PE as a candidate antigen for vaccine design upon further study. Materials and Methods: At first, the pe gene of NTHi ATCC 49766 strain (483 bp) was amplified by PCR. Then, to sequence the resulted amplicon, it was cloned into TA vector (pTZ57R/T). In the next step, the sequenced gene was sub-cloned in pBAD/gIII A vector and transformed into competent Escherichia coli TOP10. For overexpression, the recombinant bacteria were grown in broth medium containing arabinose and the recombinant protein was purified using metal affinity chromatography (Ni-nitrilotriacetic acid) (Ni-NTA agarose). Finally, the protein was detected using sodium dodecyl sulfate polyacrylamide gel electrophores (SDS-PAG) and confirmed by western blotting. Results: The cloned gene was confirmed by PCR, restriction digestion and sequencing. The sequenced gene was searched for homology in GenBank and 99% similarity was found to the already deposited genes in GenBank. Then we obtained PE using Ni-NTA agarose with up to 7 mg/mL concentration. Conclusions: The pe gene was successfully cloned and confirmed by sequencing. Finally, PE was obtained with high concentration. Due to high homology and similarity among the pe gene from NTHi ATCC 49766 and other NTHi strains in GenBank, we believe that the protein is a universal antigen to be used as a vaccine design candidate and further studies to evaluate its immunogenicity is underway. PMID:26034537

  5. No evidence of increasing Haemophilus influenzae non-b infection in Australian Aboriginal children

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    Robert I. Menzies

    2013-08-01

    Full Text Available Background. High, or increasing, rates of invasive Haemophilus influenzae (Hi type a disease have been reported from North American native children from circumpolar regions, raising the question of serotype replacement being driven by vaccination against Hi type b (Hib. Indigenous Australians from remote areas had high rates of invasive Hib disease in the past, comparable to those in North American Indigenous populations. Objective. Evaluate incidence rates of invasive Hi (overall and by serotype in Indigenous Australian children over time. Design. Descriptive study of Hi incidence rates by serotype, in the Northern Territory (NT and South Australia (SA from 2001 to 2011. Comparison of NT data with a study that was conducted in the NT in 1985–1988, before Hib vaccine was introduced. Results. The average annual rate of invasive Hi type a (Hia disease in Indigenous children aged <5 years was 11/100,000 population. Although the incidence of Hi infection in Indigenous children in 2001–2003 was lower than during 2004–2011, this may be due to changes in surveillance. No other trend over time in individual serotypes or total invasive Hi disease, in Indigenous or non-Indigenous people, was identified. Compared to 1985–1988, rates in 2001–2011 were lower in all serotype groupings, by 98% for Hib, 75% for Hia, 79% for other serotypes and 67% for non-typeable Hi. Conclusions. There is no evidence of increases in invasive disease due to Hia, other specific non-b types, or non-typeable Hi in Australian Indigenous children. These data suggest that the increase in Hia some time after the introduction of Hib vaccine, as seen in the North American Arctic Region, is not common to all populations with high pre-vaccine rates of invasive Hib disease. However, small case numbers and the lack of molecular subtyping and PCR confirmation of pre-vaccine results complicate comparisons with North American epidemiology.

  6. Nontypeable Haemophilus influenzae Clearance by Alveolar Macrophages Is Impaired by Exposure to Cigarette Smoke ▿ †

    Science.gov (United States)

    Martí-Lliteras, Pau; Regueiro, Verónica; Morey, Pau; Hood, Derek W.; Saus, Carles; Sauleda, Jaume; Agustí, Alvar G. N.; Bengoechea, José Antonio; Garmendia, Junkal

    2009-01-01

    Nontypeable Haemophilus influenzae (NTHI) is an opportunistic gram-negative pathogen that causes respiratory infections and is associated with progression of respiratory diseases. Cigarette smoke is a main risk factor for development of respiratory infections and chronic respiratory diseases. Glucocorticoids, which are anti-inflammatory drugs, are still the most common therapy for these diseases. Alveolar macrophages are professional phagocytes that reside in the lung and are responsible for clearing infections by the action of their phagolysosomal machinery and promotion of local inflammation. In this study, we dissected the interaction between NTHI and alveolar macrophages and the effect of cigarette smoke on this interaction. We showed that alveolar macrophages clear NTHI infections by adhesion, phagocytosis, and phagolysosomal processing of the pathogen. Bacterial uptake requires host actin polymerization, the integrity of plasma membrane lipid rafts, and activation of the phosphatidylinositol 3-kinase (PI3K) signaling cascade. Parallel to bacterial clearance, macrophages secrete tumor necrosis factor alpha (TNF-α) upon NTHI infection. In contrast, exposure to cigarette smoke extract (CSE) impaired alveolar macrophage phagocytosis, although NTHI-induced TNF-α secretion was not abrogated. Mechanistically, our data showed that CSE reduced PI3K signaling activation triggered by NTHI. Treatment of CSE-exposed cells with the glucocorticoid dexamethasone reduced the amount of TNF-α secreted upon NTHI infection but did not compensate for CSE-dependent phagocytic impairment. The deleterious effect of cigarette smoke was observed in macrophage cell lines and in human alveolar macrophages obtained from smokers and from patients with chronic obstructive pulmonary disease. PMID:19620348

  7. Haemophilus influenzae serotype B (Hib) seroprevalence in England and Wales in 2009.

    Science.gov (United States)

    Ladhani, Sn; Ramsay, Me; Flood, Js; Campbell, H; Slack, Mp; Pebody, R; Findlow, J; Newton, E; Wilding, M; Warrington, R; Crawford, H; Min, Sy; Gray, K; Martin, S; Frankland, S; Bokuvha, N; Laher, G; Borrow, R

    2012-01-01

    A national seroprevalence study was performed to determine the prevalence of Haemophilus influenzae type b (Hib) antibodies in England and Wales in 2009, when Hib disease incidence was the lowest ever recorded. A total of 2,693 anonymised residual sera from routine diagnostic testing submitted by participating National Health Service hospital laboratories were tested for Hib anti-polyribosyl-ribitol phosphate (PRP) IgG antibodies using a fluorescent bead assay. Median anti-PRP IgG concentrations were highest in toddlers aged 1–4 years (2.65 μg/ml), followed by children aged 5–9 years (1.95 μg/ml). Antibody concentrations were significantly lower after this age, but were still significantly higher among 10–19 year-olds (0.54 μg/ml) compared with adults aged >20 years (0.16 μg/ ml; p<0.0001). Half of the adults (51%) did not have Hib antibody concentrations ≥0.15 μg/ml, the level considered to confer short-term protection. Thus, the current excellent Hib control appears to be the result of high anti-PRP antibody concentrations in children aged up to 10 years, achieved through the various childhood vaccination campaigns offering booster immunisation. The lack of seroprotection in adults emphasises the importance of maintaining control of the disease and, most probably carriage, in children, therefore raising the question as to whether long-term routine boosting of either pre-school children or adolescents may be required. PMID:23171823

  8. The progeny of a single virgin B cell predominates the human recall B cell response to the capsular polysaccharide of Haemophilus influenzae type b

    DEFF Research Database (Denmark)

    Barington, T; Hougs, L; Juul, L;

    1996-01-01

    of Haemophilus influenzae type b coupled to tetanus toxoid. We combined affinity purification of circulating vaccine-induced Ab-secreting cells with PCR amplification of cDNA followed by cloning and sequencing. Forty-eight and 42 kappa VJ gene transcripts were analyzed from two adults, respectively. Both...

  9. Concentration and avidity of anti-Haemophilus influenzae type b (Hib) antibodies in serum samples obtained from patients for whom Hib vaccination failed

    NARCIS (Netherlands)

    Breukels, MA; Jol-van der Zijde, EM; van Tol, MJD; Rijkers, GT

    2002-01-01

    Haemophilus influenzae type b (Hib) conjugate vaccines are extremely effective in protecting infants and children from invasive Hib infections; however, vaccine failures do occur. The anti-Hib antibody production was studied both quantitatively and qualitatively in 12 patients who experienced Hib fa

  10. Incorporation of Phosphorylcholine into the Lipooligosaccharide of Nontypeable Haemophilus influenzae Does Not Correlate with the Level of Biofilm Formation In Vitro

    NARCIS (Netherlands)

    Puig, C.; Marti, S.; Hermans, P.W.M.; Jonge, M.I. de; Ardanuy, C.; Linares, J.; Langereis, J.D.

    2014-01-01

    Nontypeable Haemophilus influenzae (NTHi) is an opportunistic pathogen that causes otitis media in children and community-acquired pneumonia or exacerbations of chronic obstructive pulmonary disease in adults. A large variety of studies suggest that biofilm formation by NTHi may be an important step

  11. Increased biofilm formation by nontypeable Haemophilus influenzae isolates from patients with invasive disease or otitis media versus strains recovered from cases of respiratory infections

    NARCIS (Netherlands)

    Puig, C.; Domenech, A.; Garmendia, J.; Langereis, J.D.; Mayer, P.; Calatayud, L.; Linares, J.; Ardanuy, C.; Marti, S.

    2014-01-01

    Biofilm formation by nontypeable (NT) Haemophilus influenzae remains a controversial topic. Nevertheless, biofilm-like structures have been observed in the middle-ear mucosa of experimental chinchilla models of otitis media (OM). To date, there have been no studies of biofilm formation in large coll

  12. Characterization of the antibody response to a Haemophilus influenzae type b conjugate vaccine in children with recurrent lower respiratory tract infection

    DEFF Research Database (Denmark)

    Kristensen, K; Barington, T; Pressler, T;

    1995-01-01

    once with a Haemophilus influenzae type b (Hib) conjugate vaccine. Total IgG subclasses, total antipolysaccharide Hib antibodies, and antipolysaccharide Hib antibodies of IgM, IgG, IgA, and IgG1-4 specificity were determined by ELISA. There were no significant differences between the two groups in any...

  13. Immunogenicity of a 2-dose priming and booster vaccination with the 10-valent pneumococcal nontypeable Haemophilus influenzae protein D conjugate vaccine

    DEFF Research Database (Denmark)

    Silfverdal, Sven Arne; Høgh, Birthe; Bergsaker, Marianne Riise;

    2009-01-01

    BACKGROUND: The immunogenicity of the 10-valent pneumococcal nontypeable Haemophilus influenzae protein D-conjugate vaccine (PHiD-CV) was determined following a simplified 2-dose priming and the more commonly employed 3-dose priming both followed by a booster dose. METHODS: A total of 351 healthy...

  14. A simplification of the enzyme-linked immunospot technique. Increased sensitivity for cells secreting IgG antibodies to Haemophilus influenzae type b capsular polysaccharide

    DEFF Research Database (Denmark)

    Barington, T; Sparholt, S; Juul, L;

    1992-01-01

    A simplified enzyme-linked immunospot (ELISPOT) technique is described for the detection of cells secreting antibodies to tetanus toxoid (TT), diphtheria toxoid (DT) or Haemophilus influenzae type b capsular polysaccharide (PRP). By combining the cell suspension with the enzyme-linked secondary...

  15. Non-capsulated and capsulated Haemophilus influenzae in children with acute otitis media in Venezuela: a prospective epidemiological study

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    Naranjo Laura

    2012-02-01

    Full Text Available Abstract Background Non-typeable Haemophilus influenzae (NTHi and Streptococcus pneumoniae are major causes of bacterial acute otitis media (AOM. Data regarding AOM are limited in Latin America. This is the first active surveillance in a private setting in Venezuela to characterize the bacterial etiology of AOM in children Methods Between December 2008 and December 2009, 91 AOM episodes (including sporadic, recurrent and treatment failures were studied in 87 children enrolled into a medical center in Caracas, Venezuela. Middle ear fluid samples were collected either by tympanocentesis or spontaneous otorrhea swab sampling method. Standard laboratory and microbiological techniques were used to identify bacteria and test for antimicrobial resistance. The results were interpreted according to Clinical Laboratory Standards Institute (CLSI 2009 for non-meningitis isolates. All statistical analyses were performed using SAS 9.1 and Microsoft Excel (for graphical purposes. Results Overall, bacteria were cultured from 69.2% (63 of the 91 episodes; at least one pathogen (S. pneumoniae, H. influenzae, S. pyogenes or M. catarrhalis was cultured from 65.9% (60/91 of episodes. H. influenzae (55.5%; 35/63 episodes and S. pneumoniae (34.9%; 22/63 episodes were the most frequently reported bacteria. Among H. influenzae isolates, 62.9% (22/35 episodes were non-capsulated (NTHi and 31.4% (11/35 episodes were capsulated including types d, a, c and f, across all age groups. Low antibiotic resistance for H. influenzae was observed to amoxicillin/ampicillin (5.7%; 2/35 samples. NTHi was isolated in four of the six H. influenzae positive samples (66.7% from recurrent episodes. Conclusions We found H. influenzae and S. pneumoniae to be the main pathogens causing AOM in Venezuela. Pneumococcal conjugate vaccines with efficacy against these bacterial pathogens may have the potential to maximize protection against AOM.

  16. Antimicrobial resistance among invasive Haemophilus influenzae strains: results of a Brazilian study carried out from 1996 through 2000

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    Casagrande S.T.

    2002-01-01

    Full Text Available A total of 1712 strains of Haemophilus influenzae isolated from patients with invasive diseases were obtained from ten Brazilian states from 1996 to 2000. ß-Lactamase production was assessed and the minimum inhibitory concentrations (MIC of ampicillin, chloramphenicol, ceftriaxone and rifampin were determined using a method for broth microdilution of Haemophilus test medium. The prevalence of strains producing ß-lactamase ranged from 6.6 to 57.7%, with an overall prevalence of 18.4%. High frequency of ß-lactamase-mediated ampicillin resistance was observed in Distrito Federal (25%, São Paulo (21.7% and Paraná (18.5%. Of the 1712 strains analyzed, none was ß-lactamase negative, ampicillin resistant. A total of 16.8% of the strains were resistant to chloramphenicol, and 13.8% of these also presented resistance to ampicillin, and only 3.0% were resistant to chloramphenicol alone. All strains were susceptible to ceftriaxone and rifampin and the MIC90 were 0.015 µg/ml and 0.25 µg/ml, respectively. Ceftriaxone is the drug of choice for empirical treatment of bacterial meningitis in pediatric patients who have not been screened for drug susceptibility. The emergence of drug resistance is a serious challenge for the management of invasive H. influenzae disease, which emphasizes the fundamental role of laboratory-based surveillance for antimicrobial resistance.

  17. Haemophilus influenzae type b and its vaccines%b型流感嗜血杆菌及其疫苗

    Institute of Scientific and Technical Information of China (English)

    尹珊珊; 高正伦; 刘建凯; 郑海发

    2014-01-01

    b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)是导致婴幼儿严重细菌性感染的主要致病菌之一.Hib疫苗的研发和使用大大降低了Hib疾病的发病率.研究显示,Hib结合疫苗可有效预防婴幼儿侵袭性Hib疾病,且具有良好的安全性.此文就Hib的病原学、流行病学及疫苗研究进行综述.%Haemophilus influenzae type b (Hib) is one of the major causes of severe bacterial infections in infants and young children.With development and usage of Hib vaccines,incidence rate of Hib diseases reduces significantly.Researches show that Hib conjugate vaccine can effectively prevent invasive Hib diseases in infants and young children and has good safety.This article reviews etiology,epidemiology and vaccine research of Hib.

  18. Hydrogen peroxide scavenging is not a virulence determinant in the pathogenesis of Haemophilus influenzae type b strain Eagan

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    Van Beeumen Jozef J

    2006-01-01

    Full Text Available Abstract Background A potentially lethal flux of hydrogen peroxide (H2O2 is continuously generated during aerobic metabolism. It follows that aerobic organisms have equipped themselves with specific H2O2 dismutases and H2O2 reductases, of which catalase and the alkyl hydroperoxide reductase (AhpR are the best-studied prokaryotic members. The sequenced Haemophilus influenzae Rd genome reveals one catalase, designated HktE, and no AhpR. However, Haemophilus influenzae type b strain Eagan (Hib, a causative agent of bacterial sepsis and meningitis in young children, disrupted in its hktE gene is not attenuated in virulence, and retains the ability to rapidly scavenge H2O2. This redundancy in H2O2-scavenging is accounted for by peroxidatic activity which specifically uses glutathione as the reducing substrate. Results We show here that inside acatalasaemic H. influenzae all of the residual peroxidatic activity is catalyzed by PGdx, a hybrid peroxiredoxin-glutaredoxin glutathione-dependent peroxidase. In vitro kinetic assays on crude hktE- pgdx- H. influenzae Rd extracts revealed the presence of NAD(PH:peroxide oxidoreductase activity, which, however, appears to be physiologically insignificant because of its low affinity for H2O2 (Km = 1.1 mM. Hydroperoxidase-deficient hktE- pgdx- H. influenzae Rd showed a slightly affected aerobic growth phenotype in rich broth, while, in chemically defined medium, growth was completely inhibited by aerobic conditions, unless the medium contained an amino acid/vitamin supplement. To study the role of PGdx in virulence and to assess the requirement of H2O2-scavenging during the course of infection, both a pgdx single mutant and a pgdx/hktE double mutant of Hib were assayed for virulence in an infant rat model. The ability of both mutant strains to cause bacteremia was unaffected. Conclusion Catalase (HktE and a sole peroxidase (PGdx account for the majority of scavenging of metabolically generated H2O2 in the H

  19. Complete Deletion of the Fucose Operon in Haemophilus influenzae Is Associated with a Cluster in Multilocus Sequence Analysis-Based Phylogenetic Group II Related to Haemophilus haemolyticus: Implications for Identification and Typing.

    Science.gov (United States)

    de Gier, Camilla; Kirkham, Lea-Ann S; Nørskov-Lauritsen, Niels

    2015-12-01

    Nonhemolytic variants of Haemophilus haemolyticus are difficult to differentiate from Haemophilus influenzae despite a wide difference in pathogenic potential. A previous investigation characterized a challenging set of 60 clinical strains using multiple PCRs for marker genes and described strains that could not be unequivocally identified as either species. We have analyzed the same set of strains by multilocus sequence analysis (MLSA) and near-full-length 16S rRNA gene sequencing. MLSA unambiguously allocated all study strains to either of the two species, while identification by 16S rRNA sequence was inconclusive for three strains. Notably, the two methods yielded conflicting identifications for two strains. Most of the "fuzzy species" strains were identified as H. influenzae that had undergone complete deletion of the fucose operon. Such strains, which are untypeable by the H. influenzae multilocus sequence type (MLST) scheme, have sporadically been reported and predominantly belong to a single branch of H. influenzae MLSA phylogenetic group II. We also found evidence of interspecies recombination between H. influenzae and H. haemolyticus within the 16S rRNA genes. Establishing an accurate method for rapid and inexpensive identification of H. influenzae is important for disease surveillance and treatment.

  20. Quantitative fucK gene polymerase chain reaction on sputum and nasopharyngeal secretions to detect Haemophilus influenzae pneumonia.

    Science.gov (United States)

    Abdeldaim, Guma M K; Strålin, Kristoffer; Olcén, Per; Blomberg, Jonas; Mölling, Paula; Herrmann, Björn

    2013-06-01

    A quantitative polymerase chain reaction (PCR) for the fucK gene was developed for specific detection of Haemophilus influenzae. The method was tested on sputum and nasopharyngeal aspirate (NPA) from 78 patients with community-acquired pneumonia (CAP). With a reference standard of sputum culture and/or serology against the patient's own nasopharyngeal isolate, H. influenzae etiology was detected in 20 patients. Compared with the reference standard, fucK PCR (using the detection limit 10(5) DNA copies/mL) on sputum and NPA showed a sensitivity of 95.0% (19/20) in both cases, and specificities of 87.9% (51/58) and 89.5% (52/58), respectively. In a receiver operating characteristic curve analysis, sputum fucK PCR was found to be significantly superior to sputum P6 PCR for detection of H. influenzae CAP. NPA fucK PCR was positive in 3 of 54 adult controls without respiratory symptoms. In conclusion, quantitative fucK real-time PCR provides a sensitive and specific identification of H. influenzae in respiratory secretions. PMID:23541117

  1. The Expression of Soluble and Active Recombinant Haemophilus influenzae IgA1 Protease in E. coli

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    Shinong Long

    2010-01-01

    Full Text Available Immunoglobulin A1 (IgA1 proteases from Haemophilus influenzae are extracellular proteases that specifically cleave the hinge region of human IgA1, the predominant class of immunoglobulin present on mucosal membranes. The IgA1 proteases may have the potential to cleave IgA1 complexes in the kidney and be a therapeutic agent for IgA1 nephropathy (IgAN, a disease characterized by deposition of the IgA1 antibody in the glomerulus. We have screened for the expression of recombinant H. influenzae IgA1 protease by combining various expression plasmids, IgA1 protease constructs, and E. coli strains under multiple conditions. Using the method we have developed, approximately 20–40 mg/L of soluble and active H. influenzae IgA1 protease can be produced from E. coli strain C41(DE3, a significant increase in yield compared to the yield upon expression in H. influenzae or other related bacteria.

  2. Identification of immunogenic outer membrane proteins of Haemophilus influenzae type b in the infant rat model system

    Energy Technology Data Exchange (ETDEWEB)

    Hansen, E.J.; Frisch, C.F.; McDade, R.L. Jr.; Johnston, K.H.

    1981-06-01

    Outer membrane proteins of Haemophilus influenzae type b which are immunogenic in infant rats were identified by a radioimmunoprecipitation method. Intact cells of H. influenzae type b were radioiodinated by a lactoperoxidase-catalyzed procedure, and an outer membrane-containing fraction was prepared from these cells. These radioiodinated outer membranes were mixed with sera obtained from rats convalescing from systemic H. influenzae type b disease induced at 6 days of age, and the resultant (antibody-outer membrane protein antigen) complexes were extracted from these membranes by treatment with nonionic detergent and ethylenediaminetetraacetic acid. These soluble antibody-antigen complexes were isolated by means of adsorption to protein A-bearing staphylococci, and the radioiodinated protein antigens were identified by gel electrophoresis followed by autoradiography. Infant rats were shown to mount a readily detectable antibody response to several different proteins present in the outer membrane of H. influenzae type b. Individual infant rats were found to vary both qualitatively and quantitatively in their immune response to these immunogenic outer membrane proteins.

  3. Detection of Haemophilus influenzae by multiplex polymerase chain reaction method%多重聚合酶链反应检测流感嗜血杆菌

    Institute of Scientific and Technical Information of China (English)

    田国忠; 邵祝军; 张砺; 李晓静; 朱兵清; 杨亚静; 徐丽; 高源; 王晓蕾

    2008-01-01

    Objective To develop a rapid method for detecting Haemophilus influenzae by multiplex polymerase chain reaction (M-PCR). Methods Primers (Hi) were designed for amplification of p6 gene coding P6 protein of Haemophilus influenzae, which was used to identify Haemophilus influenzae species. Primers (Hi-cap) were designed for amplification of bexA gene which coding capsular polysaccharide (cap) synthesis was used for detecting whether Haemophilus influenzae isolates possess bexA gene relating to cap synthesis. Twelve primers (Hia-Hif) were designed for amplification of cap synthesis gene to identify the cap-type of Haemophilus influenzae. Other relative enteric pathogenic bacteria were amplified by M-PCR to serve as controls. 200 strains isolated from patients were identified.Results from M-PCR were compared to two methods including V and X factors grow requirement test and standard slide agglutination serotyping (SAST). Results The results indicated that the M-PCR assay was high specificity and sensitivity and might be valuable for differential diagnosis of Haemophilus influenzae.The sensitivity of detection was 0. 935 pg. 189 strains out of the 200 belonged to Haemophilus influenzae isolates, and one isolate was cap-type f. An agreement results were seen among the V and X factors grow requirement test, SAST and M-PCR methods. Conclusion M-PCR method showed satisfactory sensitivity, specificity and stability for detecting and identifying Haemophilus influenzae ,and could be used in clinic diagnosis, surveillance and rapid diagnosis for plague of Haemophilus influenzae.%目的 建立检测流感嗜血杆菌的多重聚合酶链反应(M-PCR)方法 .方法 合成扩增流感嗜血杆菌编码P6外膜蛋白基因的引物(Hi),鉴定流感嗜血杆菌种特异性;合成扩增流感嗜血杆菌编码荚膜基因的引物(Hi-cap),鉴定菌株是否具有荚膜;设计并合成6对扩增流感嗜血杆菌不同血清型(荚膜型)编码摹因的引物(Hia-Hif),鉴定菌株的

  4. Nonencapsulated Streptococcus pneumoniae causes otitis media during single-species infection and during polymicrobial infection with nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Murrah, Kyle A; Pang, Bing; Richardson, Stephen; Perez, Antonia; Reimche, Jennifer; King, Lauren; Wren, John; Swords, W Edward

    2015-07-01

    Streptococcus pneumoniae strains lacking capsular polysaccharide have been increasingly reported in carriage and disease contexts. Since most cases of otitis media involve more than one bacterial species, we aimed to determine the capacity of a nonencapsulated S. pneumoniae clinical isolate to induce disease in the context of a single-species infection and as a polymicrobial infection with nontypeable Haemophilus influenzae. Using the chinchilla model of otitis media, we found that nonencapsulated S. pneumoniae colonizes the nasopharynx following intranasal inoculation, but does not readily ascend into the middle ear. However, when we inoculated nonencapsulated S. pneumoniae directly into the middle ear, the bacteria persisted for two weeks post-inoculation and induced symptoms consistent with chronic otitis media. During coinfection with nontypeable H. influenzae, both species persisted for one week and induced polymicrobial otitis media. We also observed that nontypeable H. influenzae conferred passive protection from killing by amoxicillin upon S. pneumoniae from within polymicrobial biofilms in vitro. Therefore, based on these results, we conclude that nonencapsulated pneumococci are a potential causative agent of chronic/recurrent otitis media, and can also cause mutualistic infection with other opportunists, which could complicate treatment outcomes. PMID:26014114

  5. Haemophilus influenzae tipo b: situação epidemiológica no Estado de Minas Gerais, Brasil, 1993 a 1997 Haemophilus influenzae type b: epidemiological situation in the State of Minas Gerais, Brazil, 1993-1997

    Directory of Open Access Journals (Sweden)

    Sybelle de Souza Castro Miranzi

    2003-10-01

    Full Text Available Entre as doenças invasivas causadas pelo Haemophilus influenzae tipo b (Hib, destacam-se, pela freqüência e gravidade, as pneumonias e as meningites. No período de 1993 a 1997, foram notificados, em Minas Gerais, 720 casos de meningites por Hib, sendo a causa mais freqüente de meningite bacteriana em menores de um ano e a segunda causa no total de meningites. Entretanto, estimou-se uma ocorrência total de 1.160 casos considerando as meningites bacterianas não especificadas. O total de casos estimados de doença invasiva por Hib parece justificar a recente inclusão da vacina no esquema básico de imunizações. O alto custo da vacina reforça a necessidade de melhorar a vigilância epidemiológica da meningite, que constitui uma das fragilidades das ações de controle desta doença.Among Haemophilus influenzae type b (Hib invasive diseases, pneumonia and meningitis are the most relevant in public health due to their frequency and severity. From 1993 to 1997, there were 720 cases of Hib meningitis in Minas Gerais State, Brazil, representing the most frequent cause of bacterial meningitis in infants (< 1 year and the second most frequent among all causes of meningitis. The total estimated cases of invasive Hib diseases thus appear to justify the recent inclusion of the vaccine in the basic immunization protocol. The vaccine's high cost reinforces the need for more precise monitoring of the etiological diagnosis of meningitis cases, representing one of the weaknesses in the prevailing epidemiological surveillance system.

  6. Impact of the Haemophilus influenzae type b vaccination program on HIB meningitis in Brazil Impacto do programa de vacinação contra meningites causadas por Haemophilus influenzae tipo b no Brasil

    Directory of Open Access Journals (Sweden)

    Sybelle de Souza Castro Miranzi

    2007-07-01

    Full Text Available This study aimed to evaluate the impact of vaccination against Haemophilus influenzae type b (HIB in Brazil on the morbidity, mortality, and case fatality of HIB meningitis, using the Ministry of Health database and population data from the Brazilian Institute of Geography and Statistics (Instituto Brasileiro de Geografia e Estatística - IBGE. Impact was evaluated through a time series analysis (1983-2002, using regression forecasting (RF by dividing the time series into two periods: (a historical (1983-1998 and (b validation (1999-2002. Impact of the vaccination was positive, although more significant for incidence and mortality than for case fatality rates.A proposta deste trabalho foi avaliar o impacto da vacinação contra Haemophilus influenzae tipo b (HIB no Brasil sobre a morbi-mortalidade e a letalidade das meningites por HIB, a partir de base de dados fornecida pelo Ministério da Saúde e as estimativas populacionais provenientes do Instituto Brasileiro de Geografia e Estatística (IBGE. Para a avaliação do impacto utilizou-se análise de tendência temporal (1983-2002, aplicando-se a técnica RF (regression forecasting, dividindo-se a série em dois períodos: (a período histórico (1983-1998 e (b período de estimação (1999-2002. O impacto da vacinação foi positivo, embora tenha se revelado mais expressivo sobre a morbi-mortalidade que sobre a letalidade.

  7. Relationship between clinical site of isolation and ability to form biofilms in vitro in nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Obaid, Najla A; Jacobson, Glenn A; Tristram, Stephen

    2015-03-01

    Nontypeable Haemophilus influenzae (NTHi) is an opportunistic pathogen associated with a range of infections, including various lower respiratory infections, otitis media, and conjunctivitis. There is some debate as to whether or not NTHi produces biofilms and, if so, whether or not this is relevant to pathogenesis. Although many studies have examined the association between in vitro biofilm formation and isolates from a specific infection type, few have made comparisons from isolates from a broad range of isolates grouped by clinical source. In our study 50 NTHi from different clinical sources, otitis media, conjunctivitis, lower respiratory tract infections in both cystic fibrosis and non-cystic fibrosis patients, and nasopharyngeal carriage, plus 10 nasopharyngeal isolates of the commensal Haemophilus haemolyticus were tested for the ability to form biofilm by using a static microtitre plate crystal violet assay. A high degree of variation in biofilm forming ability was observed across all isolates, with no statistically significant differences observed between the groups, with the exception of the isolates from conjunctivitis. These isolates had uniformly lower biofilm forming ability compared with isolates from the other groups (p < 0.005). PMID:25706230

  8. International collaborative study for establishment of the 2nd WHO International Standard for Haemophilus influenzae type b polysaccharide.

    Science.gov (United States)

    Mawas, Fatme; Burkin, Karena; Dougall, Thomas; Saydam, Manolya; Rigsby, Peter; Bolgiano, Barbara

    2015-11-01

    In this report we present the results of a collaborative study for the preparation and calibration of a replacement International Standard (IS) for Haemophilus influenzae type b polysaccharide (polyribosyl ribitol phosphate; 5-d-ribitol-(1 → 1)-β-d-ribose-3-phosphate; PRP). Two candidate preparations were evaluated. Thirteen laboratories from 9 different countries participated in the collaborative study to assess the suitability and determine the PRP content of two candidate standards. On the basis of the results from this study, Candidate 2 (NIBSC code 12/306) has been established as the 2nd WHO IS for PRP by the Expert Committee of Biological Standards of the World Health Organisation with a content of 4.904 ± 0.185mg/ampoule, as determined by the ribose assays carried out by 11 of the participating laboratories. PMID:26298195

  9. Isolation and characterization of a UV-sensitive mutator (mutB1) mutant of Haemophilus influenzae

    International Nuclear Information System (INIS)

    The mutB1 mutant of Haemophilus influenzae is very sensitive to UV radiation but only slightly sensitive to methylmethane sulfonate or N-methyl-N'-nitro-N-nitrosoguanidine. Cultures of mutB1 cells contain high numbers of spontaneous mutants and show hypermutability after exposure to the latter mutagen. Normally high-efficiency transforming markers, as well as low-efficiency ones, transform mutB1 recipients at similarly low efficiencies. Significant host cell reactivation was observed when mutB1 cells were exposed to UV-damaged phage; however, these mutants showed a decrease in phage recombination. This mutant did not degrade its DNA following exposure to UV. It is speculated that the mutB1 mutation is similar to the Escherichia coli uvrD mutation

  10. The effects of disodium cromoglycate on enhanced adherence of Haemophilus influenzae to A549 cells infected with respiratory syncytial virus.

    Science.gov (United States)

    Fukasawa, Chie; Ishiwada, Naruhiko; Ogita, Junko; Hishiki, Haruka; Kohno, Yoichi

    2009-08-01

    Nontypeable Haemophilus influenzae (NTHi) secondary infection often complicates respiratory syncytial virus (RSV) infections. Previous studies have revealed that RSV infections enhance NTHi adherence to airway epithelial cells. In this study, we investigated the effects of disodium cromoglycate (DSCG) and corticosteroids, which are frequently used for the treatment of wheezing often related to RSV infections, on the adherence of NTHi to RSV-infected A549 cells. DSCG inhibited enhanced adherence of NTHi to RSV-infected A549 cells, whereas dexamethasone (Dex) and fluticasone propionate (Fp) did not. DSCG suppressed the expression of ICAM-1, which is one of the NTHi receptors. Furthermore, DSCG exhibited an inhibitory effect on RSV infections. It is suggested that DSCG exerts an anti-RSV effect, and consequently attenuates the expression of NTHi receptors. PMID:19390482

  11. Risk of Febrile Seizures and Epilepsy After Vaccination With Diphtheria, Tetanus, Acellular Pertussis, Inactivated Poliovirus, and Haemophilus Influenzae Type b

    DEFF Research Database (Denmark)

    Sun, Yuelian; Christensen, Jakob Christensen; Hviid, Anders;

    2012-01-01

    Context Vaccination with whole-cell pertussis vaccine carries an increased risk of febrile seizures, but whether this risk applies to the acellular pertussis vaccine is not known. In Denmark, acellular pertussis vaccine has been included in the combined diphtheria-tetanus toxoids......-acellular pertussis–inactivated poliovirus– Haemophilus influenzae type b (DTaP-IPV-Hib) vaccine since September 2002. Objective To estimate the risk of febrile seizures and epilepsy after DTaP-IPV-Hib vaccination given at 3, 5, and 12 months. Design, Setting, and Participants A population-based cohort study of 378...... seizures within 0 to 7 days (0, 1-3, and 4-7 days) after each vaccination and HR of epilepsy after first vaccination in the cohort study. Relative incidence of febrile seizures within 0 to 7 days (0, 1-3, and 4-7 days) after each vaccination in the SCCS study. Results A total of 7811 children were...

  12. Influence of prevaccination immunity on the human B-lymphocyte response to a Haemophilus influenzae type b conjugate vaccine

    DEFF Research Database (Denmark)

    Barington, T; Kristensen, K; Henrichsen, J;

    1991-01-01

    The purpose of this study was to investigate whether preexisting immunity to components of a polysaccharide-protein conjugate influences the B-lymphocyte response to vaccination with the conjugate. Thirty-two healthy adults were vaccinated once or twice with a conjugate (PRP-D) consisting...... of Haemophilus influenzae type b capsular polysaccharide (PRP) and diphtheria toxoid (DT), and the response was related to the prevaccination levels of PRP and DT antibodies. Positive correlations were found between increases in plasma PRP (median, 32.0 micrograms/ml) and DT (1.14 IU/ml) antibodies and numbers...... revaccination with PRP-D, small increases in the level of PRP antibodies (median, 2.9 micrograms/ml; n = 11) were found; no significant increase in the level of DT antibodies was seen. The numbers of PRP AbSC were lower (P = 0.04) and peaked earlier (day 7) than after the first vaccination. The isotype pattern...

  13. Crystal Structure of Homoserine Transacetylase from Haemophilus Influenzae Reveals a New Family of alpha/beta-Hydrolases

    Energy Technology Data Exchange (ETDEWEB)

    Mirza,I.; Nazi, I.; Korczynska, M.; Wright, G.; Berghuis, A.

    2005-01-01

    Homoserine transacetylase catalyzes one of the required steps in the biosynthesis of methionine in fungi and several bacteria. We have determined the crystal structure of homoserine transacetylase from Haemophilus influenzae to a resolution of 1.65 A. The structure identifies this enzyme to be a member of the alpha/beta-hydrolase structural superfamily. The active site of the enzyme is located near the end of a deep tunnel formed by the juxtaposition of two domains and incorporates a catalytic triad involving Ser143, His337, and Asp304. A structural basis is given for the observed double displacement kinetic mechanism of homoserine transacetylase. Furthermore, the properties of the tunnel provide a rationale for how homoserine transacetylase catalyzes a transferase reaction vs. hydrolysis, despite extensive similarity in active site architecture to hydrolytic enzymes.

  14. ModA2 Phasevarion Switching in Nontypeable Haemophilus influenzae Increases the Severity of Experimental Otitis Media.

    Science.gov (United States)

    Brockman, Kenneth L; Jurcisek, Joseph A; Atack, John M; Srikhanta, Yogitha N; Jennings, Michael P; Bakaletz, Lauren O

    2016-09-01

    Several human-adapted bacterial pathogens use a phasevarion (ie, a phase-variable regulon) to rapidly and reversibly regulate the expression of many genes, which include known virulence factors, yet the influence of phasevarion-mediated regulation in pathogenesis remains poorly understood. Here we examine the impact of the nontypeable Haemophilus influenzae (NTHI) ModA2 phasevarion on pathogenesis and disease severity in a chinchilla model of experimental otitis media. Chinchillas were challenged with NTHI variant populations that were either inoculated ON and remained ON, inoculated OFF and shifted ON, or inoculated OFF and remained OFF, within the middle ear. We show that populations that shift from OFF to ON within the middle ear induce significantly greater disease severity than populations that are unable to shift. These observations support the importance of phasevarion switching in NTHI pathogenesis and the necessity to considered phasevarion regulation when developing methods to treat and prevent infection.

  15. Repair of ultraviolet-damaged transforming DNA in a mismatch repair-deficient strain of Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Bagci, H.; Stuy, J.H. (Florida State Univ., Tallahassee (USA). Dept. of Biological Science)

    1982-03-01

    Ultraviolet inactivation of Haemophilus influenzae transforming DNA followed inverse square root kinetics in both mismatch repair-proficient (hex/sup +/) and deficient (hex-1) recipients. No DNA concentration effect was seen with UV-excision repair-deficient (uvr/sup -/) strains. Low-efficiency genetic markers remained more sensitive than high-efficiency ones when they were assayed on excision repair-deficient hex/sup +/ uvr/sup -/ strains. They were equally resistant when hex/sup -/ uvr/sup -/ recipients were used. This was explained by assuming that recombinational repair of UV lesions in the donor strand and mismatch repair of the recipient strand may overlap and cause double strand interruptions. This will eliminate low-efficiency transformants.

  16. Erythromycin and azithromycin transport into Haemophilus influenzae ATCC 19418 under conditions of depressed proton motive force (delta mu H)

    Energy Technology Data Exchange (ETDEWEB)

    Capobianco, J.O.; Goldman, R.C. (Abbott Laboratories, IL (USA))

    1990-09-01

    The effect of collapsing the electrochemical proton gradient (delta mu H) on ({sup 3}H)erythromycin and ({sup 14}C)azithromycin transport in Haemophilus influenzae ATCC 19418 was studied. The proton gradient and membrane potential were determined from the distribution of (2-{sup 14}C)dimethadione and rubidium-86, respectively. delta mu H was reduced from 124 to 3 mV in EDTA-valinomycin-treated cells at 22{degrees}C with 150 mM KCl and 0.1 mM carbonyl cyanide m-chlorophenylhydrazone. During the collapse of delta mu H, macrolide uptake increased. Erythromycin efflux studies strongly suggested that this increase was not due to an energy-dependent efflux pump but was likely due to increased outer membrane permeability. These data indicated that macrolide entry was not a delta mu H-driven active transport process but rather a passive diffusion process.

  17. Antibacterial FabH Inhibitors with Mode of Action Validated in Haemophilus influenzae by in Vitro Resistance Mutation Mapping.

    Science.gov (United States)

    McKinney, David C; Eyermann, Charles J; Gu, Rong-Fang; Hu, Jun; Kazmirski, Steven L; Lahiri, Sushmita D; McKenzie, Andrew R; Shapiro, Adam B; Breault, Gloria

    2016-07-01

    Fatty acid biosynthesis is essential to bacterial growth in Gram-negative pathogens. Several small molecules identified through a combination of high-throughput and fragment screening were cocrystallized with FabH (β-ketoacyl-acyl carrier protein synthase III) from Escherichia coli and Streptococcus pneumoniae. Structure-based drug design was used to merge several scaffolds to provide a new class of inhibitors. After optimization for Gram-negative enzyme inhibitory potency, several compounds demonstrated antimicrobial activity against an efflux-negative strain of Haemophilus influenzae. Mutants resistant to these compounds had mutations in the FabH gene near the catalytic triad, validating FabH as a target for antimicrobial drug discovery.

  18. Isolation and characterization of a UV-sensitive mutator (mutB1) mutant of Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Walter, R.B.; Stuy, J.H.

    1988-06-01

    The mutB1 mutant of Haemophilus influenzae is very sensitive to UV radiation but only slightly sensitive to methylmethane sulfonate or N-methyl-N'-nitro-N-nitrosoguanidine. Cultures of mutB1 cells contain high numbers of spontaneous mutants and show hypermutability after exposure to the latter mutagen. Normally high-efficiency transforming markers, as well as low-efficiency ones, transform mutB1 recipients at similarly low efficiencies. Significant host cell reactivation was observed when mutB1 cells were exposed to UV-damaged phage; however, these mutants showed a decrease in phage recombination. This mutant did not degrade its DNA following exposure to UV. It is speculated that the mutB1 mutation is similar to the Escherichia coli uvrD mutation.

  19. Haemophilus influenzae with Non-Beta-Lactamase-Mediated Beta-Lactam Resistance: Easy To Find but Hard To Categorize

    Science.gov (United States)

    Lia, Astrid; Hannisdal, Anja; Tveten, Yngvar; Matuschek, Erika; Kahlmeter, Gunnar; Kristiansen, Bjørn-Erik

    2015-01-01

    Haemophilus influenzae is a major pathogen, and beta-lactams are first-line drugs. Resistance due to altered penicillin-binding protein 3 (rPBP3) is frequent, and susceptibility testing of such strains is challenging. A collection of 154 beta-lactamase-negative isolates with a large proportion of rPBP3 (67.5%) was used to evaluate and compare Etest (Haemophilus test medium [HTM]) and disk diffusion (EUCAST method) for categorization of susceptibility to aminopenicillins and cefuroxime, using MICs generated with broth (HTM) microdilution and clinical breakpoints from CLSI and EUCAST as the gold standards. In addition, the proficiency of nine disks in screening for the rPBP3 genotype (N526K positive) was evaluated. By Etest, both essential and categorical agreement were generally poor (<70%), with high very major errors (VME) (CLSI, 13.0%; EUCAST, 34.3%) and falsely susceptible rates (FSR) (CLSI, 87.0%; EUCAST, 88.3%) for ampicillin. Ampicillin (2 μg) with adjusted (+2 mm) zone breakpoints was superior to Etest for categorization of susceptibility to ampicillin (agreement, 74.0%; VME, 11.0%; FSR, 28.3%). Conversely, Etest was superior to 30 μg cefuroxime for categorization of susceptibility to cefuroxime (agreement, 57.1% versus 60.4%; VME, 2.6% versus 9.7%; FSR, 7.1% versus 26.8%). Benzylpenicillin (1 unit) (EUCAST screening disk) and cefuroxime (5 μg) identified rPBP3 isolates with highest accuracies (95.5% and 92.2%, respectively). In conclusion, disk screening reliably detects rPBP3 H. influenzae, but false ampicillin susceptibility is frequent with routine methods. We suggest adding a comment recommending high-dose aminopenicillin therapy or the use of other agents for severe infections with screening-positive isolates that are susceptible to aminopenicillins by gradient or disk diffusion. PMID:26354813

  20. Ocorrência de Haemophilus influenzae em crianças atendidas em creches

    Directory of Open Access Journals (Sweden)

    J. M. MARIN

    2009-01-01

    Full Text Available

    As doenças provocadas por cepas de Haemophilus influenzae tipo b (Hib, como por exemplo, conjuntivite, otite média, meningite e pericardite têm apresentado uma sensível diminuição em resposta ao uso da vacina anti Hib no esquema de vacinação nacional. No entanto, com a eliminação da colonização da nasofaringe por Hib, abre-se a possibilidade da substituição de cepas colonizadoras que apresentam este sorotipo capsular por outros sorotipos ou pela colonização por H. influenzae não tipavel (NTHi. Neste sentido, as creches representam um fator de risco para a transmissão das bactérias em função do prolongado e intenso contacto entre as crianças neste ambiente. O objetivo do presente estudo foi apresentar uma revisão atualizada sobre a colonização e transmissão de H. influenzae em crianças saudáveis que freqüentam creches. Concluiu-se que as crianças que freqüentam creches devem ser continuamente monitoradas, para se verificar a eliminação da colonização na nasofaringe por Hib ou a sua substituição por cepas de outros sorotipos ou NTHi. Palavras-chave: Haemophilus influenzae, creche, transmissão, vacina anti Hib.

  1. Antimicrobial activity of innate immune molecules against Streptococcus pneumoniae, Moraxella catarrhalis and nontypeable Haemophilus influenzae

    Directory of Open Access Journals (Sweden)

    Teufert Karen

    2004-05-01

    Full Text Available Abstract Background Despite its direct connection to the nasopharynx which harbors otitis media pathogens as part of its normal flora, the middle ear cavity is kept free of these bacteria by as yet unknown mechanisms. Respiratory mucosal epithelia, including those of the middle ear and eustachian tube, secrete antimicrobial effectors including lysozyme, lactoferrin and β defensins-1 and -2. To elucidate the role of these innate immune molecules in the normal defense and maintenance of sterility of respiratory mucosa such as that of the middle ear, we assessed their effect on the respiratory pathogens nontypeable Haemophilus influenzae (NTHi 12, Moraxella catarrhalis 035E, and Streptococcus pneumoniae 3, and 6B. Methods Two assay methods, the radial assay and the liquid broth assay, were employed for testing the antimicrobial activity of the molecules. This was done in order to minimize the possibility that the observed effects were artifacts of any single assay system employed. Also, transmission electron microscopy (TEM was employed to evaluate the effect of antimicrobial innate immune molecules on OM pathogens. For the statistical analysis of the data, Student's t-test was performed. Results Results of the radial diffusion assay showed that β defensin-2 was active against all four OM pathogens tested, while treatment with β defensin-1 appeared to only affect M. catarrhalis. The radial assay results also showed that lysozyme was quite effective against S. pneumoniae 3 and 6B and was partially bacteriostatic/bactericidal against M. catarrhalis. Lysozyme however, appeared not to affect the growth of NTHi. Thus, lysozyme seems to have a more pronounced impact on the growth of the Gram-positive S. pneumoniae as compared to that of Gram-negative pathogens. Lactoferrin on the other hand, enhanced the growth of the bacteria tested. The results of the radial assays were confirmed using liquid broth assays for antimicrobial activity, and showed that

  2. Incorporation of Phosphorylcholine into the Lipooligosaccharide of Nontypeable Haemophilus influenzae Does Not Correlate with the Level of Biofilm Formation In Vitro

    OpenAIRE

    Puig, Carmen; Marti, Sara; Hermans, Peter W. M.; de Jonge, Marien I.; Ardanuy, Carmen; Liñares, Josefina; Langereis, Jeroen D.

    2014-01-01

    Nontypeable Haemophilus influenzae (NTHi) is an opportunistic pathogen that causes otitis media in children and community-acquired pneumonia or exacerbations of chronic obstructive pulmonary disease in adults. A large variety of studies suggest that biofilm formation by NTHi may be an important step in the pathogenesis of this bacterium. The objective of this report was to determine the relationship between the presence of phosphorylcholine in the lipooligosaccharide of NTHi and the level of ...

  3. High yield fermentation of haemophilus influenzae and protein p6 expression: an approach to the production of a purified vaccine. part I

    Directory of Open Access Journals (Sweden)

    D. Zuluaga

    2012-05-01

    Full Text Available The present article describes an efficient, simple and unexpensive method for the production of high cellular densities of Haemophilus influenzae for cellular and cell free vaccine production. We present here a liquid culture-based method that had not been successfully implemented in Colombia, so far having given low yields and high costs. The kinetics, costs and production of the most promisory antigen candidates for vaccine production were evaluated in liquid and solid reference media.

  4. Origin of the Diversity in DNA Recognition Domains in Phasevarion Associated modA Genes of Pathogenic Neisseria and Haemophilus influenzae

    OpenAIRE

    Gawthorne, Jayde A.; Beatson, Scott A.; Srikhanta, Yogitha N.; Fox, Kate L.; Jennings, Michael P.

    2012-01-01

    Phase variable restriction-modification (R-M) systems have been identified in a range of pathogenic bacteria. In some it has been demonstrated that the random switching of the mod (DNA methyltransferase) gene mediates the coordinated expression of multiple genes and constitutes a phasevarion (phase variable regulon). ModA of Neisseria and Haemophilus influenzae contain a highly variable, DNA recognition domain (DRD) that defines the target sequence that is modified by methylation and is used ...

  5. Influence of four modes of administration on penetration of aztreonam, cefuroxime, and ampicillin into interstitial fluid and fibrin clots and on in vivo efficacy against Haemophilus influenzae.

    OpenAIRE

    Lavoie, G Y; Bergeron, M G

    1985-01-01

    The extravascular penetration and bactericidal activity of aztreonam, cefuroxime, and ampicillin against beta-lactamase-positive and -negative Haemophilus influenzae strains were compared in a rabbit model. All groups of animals received an identical total dose of 100 mg of either antibiotic per kg given by four different intravenous modes of administration including a single large injection, four intermittent injections, a continuous infusion, and an injection followed by an infusion. Aztreo...

  6. Evidence against the reversion of mutation in the Haemophilus influenzae phage HP1c1 by preinfection treatment of host cells or phage with MNNG

    Energy Technology Data Exchange (ETDEWEB)

    Boling, M.E.; Kimball, R.F.

    1978-01-01

    N-Methyl-N'-nitro-N-nitrosoguanidine (MNNG) causes reversion of a temperature-sensitive mutation in a bacteriophage of Haemophilus influenzae if exposure to the mutagen takes place after infection but before lysis. However, neither pre-infection treatment of the phage DNA, host cells, or both will cause reversion. The reasons for this are discussed in relation to the somewhat different results in the Escherichia coli lambda phage system and in relation to error-prone repair and replication processes.

  7. Incidência de meningite por Haemophilus influenzae no RS 1999-2010: impacto da cobertura vacinal Incidence of meningitis caused by Haemophilus influenzae in the state of Rio Grande do Sul 1999-2010: impact of vaccination campaign

    Directory of Open Access Journals (Sweden)

    Marli Matiko Anraku de Campos

    2013-05-01

    Full Text Available O objetivo deste artigo é analisar e verificar a situação epidemiológica das meningites causadas pelo agente Haemophilus influenzae tipo b nos últimos 10 anos no Rio Grande do Sul. Estudo retrospectivo, descritivo, utilizando o sistema de dados de notificação de meningites, e cobertura vacinal, armazenados em base on line Tabnet - Tabulação de dados Epidemiológicos - CEVS/SES/RS, abrangendo o período de 1999 a 2010. Foram utilizados casos notificados e confirmados, tendo como critério de seleção o ano de inicio dos sintomas, idade, diagnostico e evolução. Foi analisado o Estado do Rio Grande do Sul, representado por 19 coordenadorias de saúde. Comparações entre proporções foram avaliadas pelo teste de z. No RS foram notificados 3043 casos confirmados de meningite bacteriana, sendo 6,77% dos casos causados por H. influenzae. O coeficiente de incidência da meningite por H. influenzae, sem considerar faixa etária, caiu significativamente (95,6% após 1999, assim como a mortalidade. Crianças menores de um ano continuam sendo as mais acometidas (52%, não havendo alteração na letalidade. Os resultados apresentados revelaram um impacto positivo das estratégias de vacinação contra Hib no Estado do Rio Grande do Sul nos últimos dez anos.This article seeks to analyze and update the epidemiological situation of meningitis caused by Haemophilus influenzae type b in the past 10 years in the state of Rio Grande do Sul (RS. It is a retrospective, descriptive study, which used the data notification system of meningitis and vaccination campaign coverage, stored in the Epidemiological TABNET online database, for the period from 1999 to 2010. Cases notified and confirmed were used and the selection criteria were the year when the symptoms were detected, age, diagnosis, and evolution. Nineteen health centers in the state of Rio Grande do Sul were analyzed. The z-test was used to evaluate comparisons between the proportions. In the

  8. A plasmid carrying mucA and mucB genes from pKM101 in Haemophilus influenzae and Escherichia coli

    Energy Technology Data Exchange (ETDEWEB)

    Spikes, D.; Setlow, J.K. (Brookhaven National Lab., Upton, NY (USA))

    1989-10-01

    The plasmid pMucAMucB, constructed from the Haemophilus influenzae vector pDM2, and a similar plasmid, constructed from pBR322, increased the survival after UV irradiation of Escherichia coli AB1157 with the umu-36 mutation and also caused UV-induced mutation in the E. coli strain. In H. influenzae, pMucAMucB caused a small but reproducible increase in survival after UV irradiation in wild-type cells and in a rec-1 mutant, but there was no increase in spontaneous mutation in the wild type or in the rec-1 mutant and no UV-induced mutation.

  9. A functional tonB gene is required for both virulence and competitive fitness in a chinchilla model of Haemophilus influenzae otitis media

    Directory of Open Access Journals (Sweden)

    Morton Daniel J

    2012-06-01

    Full Text Available Abstract Background Haemophilus influenzae requires heme for aerobic growth and possesses multiple mechanisms to obtain this essential nutrient. Methods An insertional mutation in tonB was constructed and the impact of the mutation on virulence and fitness in a chinchilla model of otitis media was determined. The tonB insertion mutant strain was significantly impacted in both virulence and fitness as compared to the wildtype strain in this model. Conclusions The tonB gene of H. influenzae is required for the establishment and maintenance of middle ear infection in this chinchilla model of bacterial disease.

  10. Experimental otitis media in gerbils and chinchillas with Streptococcus pneumoniae, Haemophilus influenzae, and other aerobic and anaerobic bacteria.

    Science.gov (United States)

    Fulghum, R S; Brinn, J E; Smith, A M; Daniel, H J; Loesche, P J

    1982-05-01

    To ascertain the usefulness of Mongolian gerbils as an inbred model for otitis media, 52 Mongolian gerbils (Meriones unguiculatus, strain MONT/Tum) were compared with 26 chinchillas (Chinchilla laniger) for susceptibility to Streptococcus pneumoniae type 3. Haemophilus influenzae type b, and a polymicrobic culture including anaerobes (Streptococcus intermedius, Propionibacterium acnes, Staphylococcus epidermidis, and Corynebacterium sp.). Organisms were inoculated percutaneously into the superior chamber of the middle ear bulla. The gerbils and chinchillas shared similar susceptibilities and responses to the inoculated organisms as determined by X-ray, otoscopic, histopathological, and microbiological determinations at 5 to 7 days. Koch's postulate studies proved the role of S. pneumoniae and H. influenzae in the pathology found in both animal models. The animals were also susceptible to the polymicrobic culture, although the relative virulence of the individual members of this mixture was low, suggesting that these species potentiated as a polymicrobic mixture. The Corynebacterium sp. appeared to elicit the greatest histopathological response in chronic (8-week) studies in gerbils. The gerbils were found to be useful as an alternative animal model for the study of otitis media of bacterial etiology.

  11. Haemophilus influenzae Type f Hijacks Vitronectin Using Protein H To Resist Host Innate Immunity and Adhere to Pulmonary Epithelial Cells.

    Science.gov (United States)

    Al-Jubair, Tamim; Mukherjee, Oindrilla; Oosterhuis, Sharon; Singh, Birendra; Su, Yu-Ching; Fleury, Christophe; Blom, Anna M; Törnroth-Horsefield, Susanna; Riesbeck, Kristian

    2015-12-15

    The incidence of invasive Haemophilus influenzae type b (Hib) disease has significantly decreased since the introduction of an efficient vaccine against Hib. However, in contrast to Hib, infections caused by H. influenzae serotype f (Hif) are emerging. We recently did a whole genome sequencing of an invasive Hif isolate, and reported that Hif interacts with factor H by expressing protein H (PH). In this study, upon screening with various human complement regulators, we revealed that PH is also a receptor for vitronectin (Vn), an abundant plasma protein that regulates the terminal pathway of the human complement system in addition to being a component of the extracellular matrix. Bacterial Vn binding was significantly reduced when the lph gene encoding PH was deleted in an invasive Hif isolate. The dissociation constant (KD) of the interaction between recombinant PH and Vn was 2.2 μM, as revealed by Biolayer interferometry. We found that PH has different regions for simultaneous interaction with both Vn and factor H, and that it recognized the C-terminal part of Vn (aa 352-362). Importantly, PH-dependent Vn binding resulted in better survival of the wild-type Hif or PH-expressing Escherichia coli when exposed to human serum. Finally, we observed that PH mediated an increased bacterial adherence to alveolar epithelial cells in the presence of Vn. In conclusion, our study reveals that PH most likely plays an important role in Hif pathogenesis by increasing serum resistance and adhesion to the airways. PMID:26538390

  12. Non-typeable Haemophilus influenzae protects human airway epithelial cells from a subsequent respiratory syncytial virus challenge.

    Science.gov (United States)

    Hartwig, Stacey M; Ketterer, Margaret; Apicella, Michael A; Varga, Steven M

    2016-11-01

    Respiratory syncytial virus (RSV) and the common commensal and opportunistic pathogen, non-typeable Haemophilus influenzae (NTHi) both serve as a frequent cause of respiratory infection in children. Although it is well established that some respiratory viruses can increase host susceptibility to secondary bacterial infections, few studies have examined how commensal bacteria could influence a secondary viral response. Here, we examined the impact of NTHi exposure on a subsequent RSV infection of human bronchial epithelial cells (16HBE14o-). Co-culture of 16HBE14o- cells with NTHi resulted in inhibition of viral gene expression following RSV infection. 16HBE14o- cells co-cultured with heat-killed NTHi failed to protect against an RSV infection, indicating that protection requires live bacteria. However, NTHi did not inhibit influenza A virus replication, indicating that NTHi-mediated protection was RSV-specific. Our data demonstrates that prior exposure to a commensal bacterium such as NTHi can elicit protection against a subsequent RSV infection.

  13. Oxygen-independent inactivation of Haemophilus influenzae transforming DNA by monochromatic radiation: action spectrum, effect of histidine and repair

    Energy Technology Data Exchange (ETDEWEB)

    Cabrera-Juarez, E.; Setlow, J.K.; Swenson, P.A.; Peak, M.J.

    1976-01-01

    The action spectrum for the oxygen-independent inactivation of native transforming DNA from Haemophilus influenzae with near-uv radiation revealed a shoulder beginning at 334 and extending to 460 nm. The presence of 0.2 M histidine during irradiation produced a small increase in inactivation at 254, 290 and 313 nm, a large increase at 334 nm and a decrease in inactivation at 365, 405, and 460 nm. Photoreactivation did not reverse the DNA damage produced at pH 7.0 at 334, 365, 405 and 460 nm, but did reactivate the DNA after irradiation at 254, 290 and 313 nm. The inactivation of DNA irradiated at 254, 290 and 313 nm was considerably greater when the transforming ability was assayed in an excision-defective mutant compared with the wild type, although DNA irradiated at 334, 365, 405 and 460 nm showed smaller differences. These results suggest that the oxygen-independent inactivation of H. influenzae DNA at pH 7 by irradiation at 334, 365, 405 and 460 nm is caused by lesions other than pyrimidine dimers.

  14. Mutagenesis and ultraviolet inactivation of transforming DNA of ``Haemophilus influenzae`` complexed with a ``Bacillus subtilis`` protein that alter DNA conformation

    Energy Technology Data Exchange (ETDEWEB)

    Setlow, Jane K. [Brookhaven National Lab., Upton, NY (United States); Setlow, Barbara C.; Setlow, Peter [Connecticut Univ., Farmington, CT (United States)

    1996-12-31

    The wild-type ``Bacillus subtilis`` spore protein, SspC{sup wt}, binds to DNA ``in vitro`` and ``in vivo`` and changes the conformation of DNA from B to A. Synthesis of the cloned SspC{sup wt} gene in ``Escherichia coli`` also causes large increases in mutation frequency. Binding of SspC{sup wt} to transforming DNA from ``Haemophilus influenzae`` made the DNA resistant to ultraviolet (UV) radiation. The mutant protein, SspC{sup ala}, which does not bind DNA, did not change the UV resistance. The UV sensitivity of the DNA/SspC{sup wt} complex was not increased when the recipients of the DNA were defective in excision of pyrimidine dimers. These data indicate that the ``H. influenzae`` excision mechanism does not operate on the spore photoproduct formed by UV irradiation of the complex. Selection for the streptomycin- or erythromycin-resistance markers on the transforming DNA evidenced significant mutations at loci closely linked to these, but not at other loci. SspC{sup wt} apparently entered the cell attached to the transforming DNA, and caused mutations in adjacent loci. The amount of such mutations decreased when the transforming DNA was UV irradiated, because UV unlinks linked markers. (author). 22 refs, 4 figs, 4 tabs.

  15. Polyarticular Septic Arthritis Caused by Haemophilus influenzae Serotype f in an 8-Month-Old Immunocompetent Infant: A Case Report and Review of the Literature.

    Science.gov (United States)

    Ali, Raheel Ahmed; Kaplan, Sheldon L; Rosenfeld, Scott B

    2015-01-01

    Background. The standard use of vaccinations against pathogens has resulted in a decreased incidence of musculoskeletal infections caused by these previously common bacterial pathogens. Consequently, the incidence of infections caused by atypical bacteria is rising. This report presents a case of septic arthritis caused by non-type b H. influenzae in a pediatric patient. Methods. We report a case of an infant with polyarticular septic arthritis caused by H. influenzae serotype f. A literature review was conducted with the inclusion criteria of case reports and studies published between 2004 and 2013 addressing musculoskeletal H. influenzae infections. Results. An 8-month-old female presented with pain and swelling in her right ankle and left elbow. The patient was diagnosed with septic arthritis and underwent incision and drainage. Wound and blood cultures were positive for Haemophilus influenzae serotype f. In addition to treatment with IV antibiotics, the patient underwent immunocompetency studies, which were normal. Subsequent follow-up revealed eradication of the infection. Conclusions. Haemophilus influenzae non-type b may cause serious invasive infections such as sepsis or septic arthritis in children with or without predisposing factors such as immunodeficiency or asplenia. Optimal treatment includes surgical management, culture driven IV antibiotics, and an immunologic workup.

  16. Polyarticular Septic Arthritis Caused by Haemophilus influenzae Serotype f in an 8-Month-Old Immunocompetent Infant: A Case Report and Review of the Literature

    Directory of Open Access Journals (Sweden)

    Raheel Ahmed Ali

    2015-01-01

    Full Text Available Background. The standard use of vaccinations against pathogens has resulted in a decreased incidence of musculoskeletal infections caused by these previously common bacterial pathogens. Consequently, the incidence of infections caused by atypical bacteria is rising. This report presents a case of septic arthritis caused by non-type b H. influenzae in a pediatric patient. Methods. We report a case of an infant with polyarticular septic arthritis caused by H. influenzae serotype f. A literature review was conducted with the inclusion criteria of case reports and studies published between 2004 and 2013 addressing musculoskeletal H. influenzae infections. Results. An 8-month-old female presented with pain and swelling in her right ankle and left elbow. The patient was diagnosed with septic arthritis and underwent incision and drainage. Wound and blood cultures were positive for Haemophilus influenzae serotype f. In addition to treatment with IV antibiotics, the patient underwent immunocompetency studies, which were normal. Subsequent follow-up revealed eradication of the infection. Conclusions. Haemophilus influenzae non-type b may cause serious invasive infections such as sepsis or septic arthritis in children with or without predisposing factors such as immunodeficiency or asplenia. Optimal treatment includes surgical management, culture driven IV antibiotics, and an immunologic workup.

  17. Molecular characterisation of the clonal emergence of high-level ciprofloxacin-monoresistant Haemophilus influenzae in the Region of Southern Denmark

    DEFF Research Database (Denmark)

    Fuursted, Kurt; Hartmeyer, Gitte Nyvang; Stegger, Marc;

    2016-01-01

    Haemophilus influenzae is an important human pathogen usually susceptible to quinolones. Here we report the emergence of high-level ciprofloxacin-monoresistant H. influenzae in the Region of Southern Denmark. Four isolates were collected for phenotypic and molecular characterisation using whole......-genome sequencing (WGS). During an 18-month period, the occurrence of high-level ciprofloxacin-monoresistant H. influenzae in patients aged 1-77 years from sputum, ear and eye samples was detected. An epidemiological link between the patients could not be identified. The isolates were non-encapsulated, biotype III...... description of the clonal emergence of high-level monoresistant H. influenzae due to amino acid substitutions in gyrA, parC and parE....

  18. 流感嗜血杆菌感染与耐药%The infections and resistance of Haemophilus influenzae

    Institute of Scientific and Technical Information of China (English)

    王爱华

    2016-01-01

    流感嗜血杆菌(Haemophilus influenzae,Hi)是儿童时期细菌性脑膜炎、肺炎和菌血症的主要细菌性病原,其抗生素耐药性存在地区和时间差异。β-内酰胺酶是 Hi 对β-内酰胺类抗生素耐药的主要机制,抗生素使用是影响 Hi 耐药性变化的主要因素,Hi 对第三代头孢菌素仍保持较高敏感性。耐药性监测为儿童 Hi 感染抗生素应用提供依据,疫苗预防是控制 Hi 感染的未来方向。%Haemophilus influenzae (Hi)is an important human pathogen that causes severe infections inclu-ding meningitis,bacteremic pneumonia and sepsis,mostly affecting young children.The resistance of Hi to antimicro-bials has risen and wide variations exist in antibiotic resistance,both geographically and over time.The main mecha-nisms of resistance of Hi are those to β-lactam antibiotics.Antibiotic utilization is the main risk factors for the deve-lopment of antimicrobial resistance.Third -generation cephalosporins is the most effective antibiotic agent to treat the infections of Hi in children population.With continued research,progress toward a broadly effective vaccine to prevent infections caused by Hi is expected in the near future.

  19. Comparison of Established Diagnostic Methodologies and a Novel Bacterial smpB Real-Time PCR Assay for Specific Detection of Haemophilus influenzae Isolates Associated with Respiratory Tract Infections

    OpenAIRE

    Reddington, Kate; Schwenk, Stefan; Tuite, Nina; Platt, Gareth; Davar, Danesh; Coughlan, Helena; Personne, Yoann; Gant, Vanya; Enne, Virve I.; Zumla, Alimuddin; Barry, Thomas

    2015-01-01

    Haemophilus influenzae is a significant causative agent of respiratory tract infections (RTI) worldwide. The development of a rapid H. influenzae diagnostic assay that would allow for the implementation of infection control measures and also improve antimicrobial stewardship for patients is required. A number of nucleic acid diagnostics approaches that detect H. influenzae in RTIs have been described in the literature; however, there are reported specificity and sensitivity limitations for th...

  20. Adoption of Preventive Measures and Attitudes toward the H1N1 Influenza Pandemic in Schools

    Science.gov (United States)

    Pérez, Anna; Rodríguez, Tània; López, Maria José; Continente, Xavier; Nebot, Manel

    2016-01-01

    Background: This study describes the perceived impact of H1N1 influenza and the adoption of the recommended measures to address the pandemic in schools. Methods: A cross-sectional self-reported survey was conducted in 433 schools in Barcelona addressed to the school principal or the H1N1 influenza designated person. A descriptive analysis was…

  1. Advances in the research of protein D of nontypeable Haemophilus influenzae%不可分型流感嗜血杆菌蛋白D研究进展

    Institute of Scientific and Technical Information of China (English)

    陈祥鹏; 王健

    2010-01-01

    流感嗜血杆菌(Haemophilus influenzae,Hi)蛋白D(Protein D,PD)是高度保守的表面脂蛋白,相对分子质量42 000,存在于所有的可分型Hi和不可分型Hi(nontypeable Haemophilus influenzae,NTHi)中,是一种具有甘油磷酸二酯酶活性的细菌毒力因子,能导致宿主上皮细胞释放磷酸胆碱,在Hi引起呼吸道感染过程中起重要作用.以NTHi PD作为载体蛋白的肺炎链球菌多糖结合疫苗,不但能预防肺炎链球菌性耳炎,而且对NTHi引起的急性中耳炎也具有预防作用.PD是迄今为止第一种能在人类机体中诱导保护性应答的NTHi抗原,也是最具潜力的多糖结合疫苗候选载体蛋白.%Protein D (PD) is a highly conserved surface lipoprotein(M, 42 000) found in all Hae mophilus influenzae (Hi), including nontypeable Haemophilus influenzae (NTHi).PD is a virulence factor with glycerophosphodiesterase activity, which can lead to the release of phosphorylcholine from host epithelial cells, and plays an important role in the pathogenesis of respiratory tract infection caused by Hi.Pneumococcal polysaccharide conjugate vaccine using NTHi PD as a carrier protein can protect against acute otitis media caused by both pneumococci and NTHi.PD is not only a potential carrier protein candidate for polysaccharide conjugate vaccines but also the first NTHi antigen which can induce protective response in human.

  2. Live Streptococcus pneumoniae, Haemophilus influenzae, and Neisseria meningitidis activate the inflammatory response trhough Toll-like receptors 2, 4, and 9 in species-specific patterns

    DEFF Research Database (Denmark)

    Mogensen, T.H.; Paludan, Søren Riis; Kilian, Mogens;

    2006-01-01

    activation by live bacteria. Here, we demonstrate that live Streptococcus pneumoniae, Haemophilus influenzae type b, and Neisseria meningitidis, the three principal causes of bacterial meningitis, use distinct sets of TLRs to trigger the inflammatory response. Using human embryonic kidney 293 cell lines......, each overexpressing one type of TLR, we found that S. pneumoniae triggered activation of the transcription factor nuclear factor-kappaB and expression of interleukin-8, only in cells expressing TLR2 or -9. The same response was evoked by H. influenzae in cells expressing TLR2 or -4 and by N...... and confirmed the essential role of these TLRs and also identified differential functions of TLRs in activation of the inflammatory response. Collectively, we here demonstrate that S. pneumoniae, H. influenzae, and N. meningitidis each activate several TLRs in species-specific patterns and show that infection...

  3. Applying Central Composite Design and Response Surface Methodology to Optimize Growth and Biomass Production of Haemophilus influenzae Type b

    Science.gov (United States)

    Momen, Seyed Bahman; Siadat, Seyed Davar; Akbari, Neda; Ranjbar, Bijan; Khajeh, Khosro

    2016-01-01

    Background Haemophilus influenzae type b (Hib) is the leading cause of bacterial meningitis, otitis media, pneumonia, cellulitis, bacteremia, and septic arthritis in infants and young children. The Hib capsule contains the major virulence factor, and is composed of polyribosyl ribitol phosphate (PRP) that can induce immune system response. Vaccines consisting of Hib capsular polysaccharide (PRP) conjugated to a carrier protein are effective in the prevention of the infections. However, due to costly processes in PRP production, these vaccines are too expensive. Objectives To enhance biomass, in this research we focused on optimizing Hib growth with respect to physical factors such as pH, temperature, and agitation by using a response surface methodology (RSM). Materials and Methods We employed a central composite design (CCD) and a response surface methodology to determine the optimum cultivation conditions for growth and biomass production of H. influenzae type b. The treatment factors investigated were initial pH, agitation, and temperature, using shaking flasks. After Hib cultivation and determination of dry biomass, analysis of experimental data was performed by the RSM-CCD. Results The model showed that temperature and pH had an interactive effect on Hib biomass production. The dry biomass produced in shaking flasks was about 5470 mg/L, which was under an initial pH of 8.5, at 250 rpm and 35° C. Conclusions We found CCD and RSM very effective in optimizing Hib culture conditions, and Hib biomass production was greatly influenced by pH and incubation temperature. Therefore, optimization of the growth factors to maximize Hib production can lead to 1) an increase in bacterial biomass and PRP productions, 2) lower vaccine prices, 3) vaccination of more susceptible populations, and 4) lower risk of Hib infections.

  4. Regulation of virulence gene expression resulting from Streptococcus pneumoniae and nontypeable Haemophilus influenzae interactions in chronic disease.

    Directory of Open Access Journals (Sweden)

    Emily K Cope

    Full Text Available Chronic rhinosinusitis (CRS is a common inflammatory disease of the sinonasal cavity mediated, in part, by polymicrobial communities of bacteria. Recent molecular studies have confirmed the importance of Streptococcus pneumoniae and nontypeable Haemophilus influenzae (NTHi in CRS. Here, we hypothesize that interaction between S. pneumoniae and NTHi mixed-species communities cause a change in bacterial virulence gene expression. We examined CRS as a model human disease to validate these polymicrobial interactions. Clinical strains of S. pneumoniae and NTHi were grown in mono- and co-culture in a standard biofilm assay. Reverse transcriptase real-time PCR (RTqPCR was used to measure gene expression of key virulence factors. To validate these results, we investigated the presence of the bacterial RNA transcripts in excised human tissue from patients with CRS. Consequences of physical or chemical interactions between microbes were also investigated. Transcription of NTHi type IV pili was only expressed in co-culture in vitro, and expression could be detected ex vivo in diseased tissue. S. pneumoniae pyruvate oxidase was up-regulated in co-culture, while pneumolysin and pneumococcal adherence factor A were down-regulated. These results were confirmed in excised human CRS tissue. Gene expression was differentially regulated by physical contact and secreted factors. Overall, these data suggest that interactions between H. influenzae and S. pneumoniae involve physical and chemical mechanisms that influence virulence gene expression of mixed-species biofilm communities present in chronically diseased human tissue. These results extend previous studies of population-level virulence and provide novel insight into the importance of S. pneumoniae and NTHi in CRS.

  5. THE SAFETY OF HAEMOPHILUS INFLUENZAE TYPE B/POLYRIBOSYLRIBITOL PHOSPHATE-TETANUS (HIB/PRP-T VACCINE, PHASE I STUDY

    Directory of Open Access Journals (Sweden)

    Kusnandi Rusmil

    2015-09-01

    Full Text Available Objective: To assess the safety and immunogenicity of the Haemophilus influenzae type b/polyribosylribitol phosphate-Tetanus (Hib/PRP-T liquid vaccine in healthy adults. Methods: An open label prospective intervention phase I study was conducted in Dr. Hasan Sadikin General Hospital from November to December 2010. Healthy adults aged 18−40 years were eligible to participate. Participants received one dose of Hib/PRP-T liquid vaccine. Blood samples were taken before, 4 days, and 1 month after vaccination. For a 28-day period following vaccination, solicited adverse events were recorded in the subjects’ diary and assessed afterward. Results: No local reactions or immediate systemic events were observed during the 30-minute period after immunization. There were no serious local or systemic reactions in this study. All of local and systemic reactions observed were slight, transient, self-limiting, and lasting no more than 72 hours after the administration of the vaccine. These reactions resolved without any medical intervention. Hematological and biochemical indices before and 4 days after vaccination were in normal limits. All subjects reached protective levels of antibodies (seroprotectivity against Hib. All subjects demonstrated antibodies performing high bactericidal activities 1 month after immunization. Conclusions: This study demonstrates that liquid Hib/PRP-T vaccine is highly immunogenic and beneficially safe when administered to healthy adults.

  6. Cigarette Smoke-Induced Lung Disease Predisposes to More Severe Infection with Nontypeable Haemophilus influenzae: Protective Effects of Andrographolide.

    Science.gov (United States)

    Tan, W S Daniel; Peh, Hong Yong; Liao, Wupeng; Pang, Chu Hui; Chan, Tze Khee; Lau, Suk Hiang; Chow, Vincent T; Wong, W S Fred

    2016-05-27

    Cigarette smoke (CS) is associated with many maladies, one of which is chronic obstructive pulmonary disease (COPD). As the disease progresses, patients are more prone to develop COPD exacerbation episodes by bacterial infection, particularly to nontypeable Haemophilus influenza (NTHi) infection. The present study aimed to develop a CS-exposed mouse model that increases inflammation induced by NTHi challenge and investigate the protective effects of andrographolide, a bioactive molecule with anti-inflammatory and antioxidant properties isolated from the plant Andrographis paniculata. Female BALB/c mice exposed to 2 weeks of CS followed by a single intratracheal instillation of NTHi developed increased macrophage and neutrophil pulmonary infiltration, augmented cytokine levels, and heightened oxidative damage. Andrographolide effectively reduced lung cellular infiltrates and decreased lung levels of TNF-α, IL-1β, CXCL1/KC, 8-OHdG, matrix metalloproteinase-8 (MMP-8), and MMP-9. The protective actions of andrographolide on CS-predisposed NTHi inflammation might be attributable to increased nuclear factor erythroid-2-related factor 2 (Nrf2) activation and decreased Kelch-like ECH-associated protein 1 (Keap1) repressor function, resulting in enhanced gene expression of antioxidant enzymes including heme oxygenase-1 (HO-1), glutathione reductase (GR), glutathione peroxidase-2 (GPx-2), glutamate-cysteine ligase modifier (GCLM), and NAD(P)H quinone oxidoreductase 1 (NQO1). Taken together, these findings strongly support a therapeutic potential for andrographolide in preventing lung inflammation caused by NTHi in cigarette smokers. PMID:27104764

  7. A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate.

    Science.gov (United States)

    Abdelhameed, Ali Saber; Adams, Gary G; Morris, Gordon A; Almutairi, Fahad M; Duvivier, Pierre; Conrath, Karel; Harding, Stephen E

    2016-01-01

    Three important physical properties which may affect the performance of glycoconjugate vaccines against serious disease are molar mass (molecular weight), heterogeneity (polydispersity), and conformational flexibility in solution. The dilute solution behaviour of native and activated capsular polyribosylribitol (PRP) polysaccharides extracted from Haemophilus influenzae type b (Hib), and the corresponding glycoconjugate made by conjugating this with the tetanus toxoid (TT) protein have been characterized and compared using a combination of sedimentation equilibrium and sedimentation velocity in the analytical ultracentrifuge with viscometry. The weight average molar mass of the activated material was considerably reduced (Mw ~ 0.24 × 10(6) g.mol(-1)) compared to the native (Mw ~ 1.2 × 10(6) g.mol(-1)). Conjugation with the TT protein yielded large polydisperse structures (of Mw ~ 7.4 × 10(6) g.mol(-1)), but which retained the high degree of flexibility of the native and activated polysaccharide, with frictional ratio, intrinsic viscosity, sedimentation conformation zoning behaviour and persistence length all commensurate with highly flexible coil behaviour and unlike the previously characterised tetanus toxoid protein (slightly extended and hydrodynamically compact structure with an aspect ratio of ~3). This non-protein like behaviour clearly indicates that it is the carbohydrate component which mainly influences the physical behaviour of the glycoconjugate in solution. PMID:26915577

  8. An innovative method for quality control of conjugated Haemophilus influenzae vaccines: A short review of two-dimensional nanoparticle electrophoresis.

    Science.gov (United States)

    Tietz, Dietmar

    2009-12-25

    This article provides an overview of a 2D agarose electrophoretic procedure for the characterization of semi-synthetic Haemophilus influenzae type b meningitis vaccines that were prepared for the immunization of small children. The analysis of such vaccines has been particularly challenging because the vaccine particles (i) are highly negatively charged, (ii) are as large as or even larger than intact viruses, and (iii) have a continuous (polydisperse) size distribution because of randomizing steps in the vaccine production (sonification and crosslinking). As a result of these characteristics, 1D electrophoresis of the vaccines produced smears without discernable peaks, but with a second dimension of separation a characteristic vaccine fingerprint was obtained. Whereas O'Farrell gels can accomplish a 2D separation according to size and charge for samples with protein-sized particles, nondenaturing 2D agarose electrophoresis achieves a similar result for much larger virus-sized particles. The separation principle, however, is different. Even though the 2D electrophoretic method was developed from 1983 to 1995, it remains a promising tool for vaccine quality control and for predicting vaccine effectiveness. Modern technology makes the analysis significantly more practical and affordable than it was more than 10 years ago, and the method is applicable to a variety of conjugated vaccines and complex mixtures of virus-sized particles.

  9. Combined Bacteria Microarray and Quartz Crystal Microbalance Approach for Exploring Glycosignatures of Nontypeable Haemophilus influenzae and Recognition by Host Lectins.

    Science.gov (United States)

    Kalograiaki, Ioanna; Euba, Begoña; Proverbio, Davide; Campanero-Rhodes, María A; Aastrup, Teodor; Garmendia, Junkal; Solís, Dolores

    2016-06-01

    Recognition of bacterial surface epitopes by host receptors plays an important role in the infectious process and is intimately associated with bacterial virulence. Delineation of bacteria-host interactions commonly relies on the detection of binding events between purified bacteria- and host-target molecules. In this work, we describe a combined microarray and quartz crystal microbalance (QCM) approach for the analysis of carbohydrate-mediated interactions directly on the bacterial surface, thus preserving the native environment of the bacterial targets. Nontypeable Haemophilus influenzae (NTHi) was selected as a model pathogenic species not displaying a polysaccharide capsule or O-antigen-containing lipopolysaccharide, a trait commonly found in several important respiratory pathogens. Here, we demonstrate the usefulness of NTHi microarrays for exploring the presence of carbohydrate structures on the bacterial surface. Furthermore, the microarray approach is shown to be efficient for detecting strain-selective binding of three innate immune lectins, namely, surfactant protein D, human galectin-8, and Siglec-14, to different NTHi clinical isolates. In parallel, QCM bacteria-chips were developed for the analysis of lectin-binding kinetics and affinity. This novel QCM approach involves capture of NTHi on lectin-derivatized chips followed by formaldehyde fixation, rendering the bacteria an integrated part of the sensor chip, and subsequent binding assays with label-free lectins. The binding parameters obtained for selected NTHi-lectin pairs provide further insights into the interactions occurring at the bacterial surface.

  10. Degradation of DNA in Haemophilus influenzae cells after x-ray irradiation. II. Comparison with theoretical models

    Energy Technology Data Exchange (ETDEWEB)

    Randolph, M.L.

    1976-01-01

    Models of the kinetics of degradation of DNA in bacterial cells following exposure to x-rays are developed and tested using data obtained with various strains of Haemophilus influenzae cells. Different mathematical models allow for different initial distributions of DNA lengths, depending on cell phase and assumed replication model, and for unilateral or bilateral degradation from initiation points, which are taken as single-strand breaks. In order to explain the observed magnitude and time course of degradation an interruption-of-degradation probability, which may be interpreted as the result of an x-ray inducible inhibitor, is introduced. Interruption of degradation is interpreted as an early and essential, but not necessarily sufficient, step for repair of DNA. Empirical best fits to the kinetic data were obtained by iterative calculation methods based on varying the rate parameters. Depending somewhat on the strain and cell phase, the data seem best described by assuming bilateral degradation, perhaps at different rates, whose total for log phase wild-type cells is roughly 5 x 10/sup 3/ nucleotides/min and an interruption rate of about 0.03/min. For stationary phase the total degradation rate is greater and the probability of degradation unchanged. Differences in the kinetics between strains are discussed briefly.

  11. Ultraviolet sensitivity of the addition, deletion and replacement of long nonhomologous DNA segments by genetic transformation of Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Walter, R.B.; Stuy, J.H. (Florida State Univ., Tallahassee (USA). Dept. of Biological Science)

    1982-03-01

    The construction and some properties of Haemophilus influenzae Rd strains with long and different R plasmid-derived DNA segments (nonhomologous inserts) at the same site in the HP1 prophage have previously been described. These inserts can be added to a recipient's genome by genetic transformation, they can be deleted from the recipient genome, or they can be replaced by another insert. It is reported that the UV inactivation of all three phenomena followed single hit kinetics. Deletion was roughly 10 times more resistant; its UV-sensitivity equalled that of a high-efficiency point mutation. There was an inverse correlation between UV-sensitivity and additive transformation efficiency of the various inserts; sensitivity may thus be a measure of insert size. This correlation was not seen for deletion. All three phenomena were more sensitive when they were measured on excision repair-deficient uvr/sup -/ recipients. The dose-reduction factor for addition was about 1.5 while it was about 2.6 for deletion.

  12. The ToxAvapA toxin-antitoxin locus contributes to the survival of nontypeable Haemophilus influenzae during infection.

    Directory of Open Access Journals (Sweden)

    Dabin Ren

    Full Text Available Nontypeable Haemophilus influenzae (NTHi is an opportunistic pathogen that is a common cause of acute and recurrent mucosal infections. One uncharacterized NTHi toxin-antitoxin (TA module, NTHI1912-1913, is a host inhibition of growth (higBA homologue. We hypothesized that this locus, which we designated toxAvapA, contributed to NTHi survival during infection. We deleted toxAvapA and determined that growth of the mutant in defined media was not different from the parent strain. We tested the mutant for persistence during long-term in vitro co-culture with primary human respiratory tissues, which revealed that the ΔtoxAvapA mutant was attenuated for survival. We then performed challenge studies using the chinchilla model of otitis media and determined that mutant survival was also reduced in vivo. Following purification, the toxin exhibited ribonuclease activity on RNA in vitro, while the antitoxin did not. A microarray comparison of the transcriptome revealed that the tryptophan biosynthetic regulon was significantly repressed in the mutant compared to the parent strain. HPLC studies of conditioned medium confirmed that there was no significant difference in the concentration of tryptophan remaining in the supernatant, indicating that the uptake of tryptophan by the mutant was not affected. We conclude that the role of the NTHi toxAvapA TA module in persistence following stress is multifactorial and includes effects on essential metabolic pathways.

  13. Structure of a small-molecule inhibitor complexed with GlmU from Haemophilus influenzae reveals an allosteric binding site

    Energy Technology Data Exchange (ETDEWEB)

    Mochalkin, Igor; Lightle, Sandra; Narasimhan, Lakshmi; Bornemeier, Dirk; Melnick, Michael; VanderRoest, Steven; McDowell, Laura (Pfizer)

    2008-04-02

    N-Acetylglucosamine-1-phosphate uridyltransferase (GlmU) is an essential enzyme in aminosugars metabolism and an attractive target for antibiotic drug discovery. GlmU catalyzes the formation of uridine-diphospho-N-acetylglucosamine (UDP-GlcNAc), an important precursor in the peptidoglycan and lipopolisaccharide biosynthesis in both Gram-negative and Gram-positive bacteria. Here we disclose a 1.9 {angstrom} resolution crystal structure of a synthetic small-molecule inhibitor of GlmU from Haemophilus influenzae (hiGlmU). The compound was identified through a high-throughput screening (HTS) configured to detect inhibitors that target the uridyltransferase active site of hiGlmU. The original HTS hit exhibited a modest micromolar potency (IC{sub 50} - 18 {mu}M in a racemic mixture) against hiGlmU and no activity against Staphylococcus aureus GlmU (saGlmU). The determined crystal structure indicated that the inhibitor occupies an allosteric site adjacent to the GlcNAc-1-P substrate-binding region. Analysis of the mechanistic model of the uridyltransferase reaction suggests that the binding of this allosteric inhibitor prevents structural rearrangements that are required for the enzymatic reaction, thus providing a basis for structure-guided design of a new class of mechanism-based inhibitors of GlmU.

  14. Post-GAVI sustainability of the Haemophilus influenzae type b vaccine program: The potential role of economic evaluation.

    Science.gov (United States)

    Le, Phuc; Nghiem, Van T; Swint, J Michael

    2016-09-01

    Haemophilus influenzae type b (Hib) can cause severe invasive diseases which are, however, preventable by vaccination. To increase access to Hib vaccine, GAVI - the Vaccine Alliance - has provided financial support for 73 lower income countries worldwide. At the same time, GAVI has been implementing its co-financing policy, requiring recipient countries to pay a portion of vaccine costs and to increase this amount over time. Starting in 2016, 5 countries will stop receiving GAVI funding and procure the vaccine themselves. Although the graduating countries have access to the UNICEF/GAVI tendered vaccine price for 5 more years, the uncertainty in market vaccine price may hamper the post-GAVI program sustainability. A possible increase in vaccine price would cause a significant burden on governmental budgets, discouraging countries to continue the program. As a special tool, economic evaluation (EE) can assist decision makers by identifying the maximum affordable vaccine price for countries to pay. Given that only 6 GAVI-eligible countries have such analyses published, more EEs are necessary to strengthen countries' commitment during this transition period. The information will also be useful for manufacturers to determine their pricing policy.

  15. Intercellular adhesion molecule 1 serves as a primary cognate receptor for the Type IV pilus of nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Novotny, Laura A; Bakaletz, Lauren O

    2016-08-01

    Nontypeable Haemophilus influenzae (NTHI) utilizes the Type IV pilus (Tfp) to adhere to respiratory tract epithelial cells thus colonizing its human host; however, the host cell receptor to which this adhesive protein binds is unknown. From a panel of receptors engaged by Tfp expressed by other bacterial species, we showed that the majority subunit of NTHI Tfp, PilA, bound to intercellular adhesion molecule 1 (ICAM1) and that this interaction was both specific and of high affinity. Further, Tfp-expressing NTHI inoculated on to polarized respiratory tract epithelial cells that expressed ICAM1 were significantly more adherent compared to Tfp-deficient NTHI or NTHI inoculated on to epithelial cells to which ICAM1 gene expression was silenced. Moreover, pre-incubation of epithelial cells with recombinant soluble PilA (rsPilA) blocked adherence of NTHI, an outcome that was abrogated by admixing rsPilA with ICAM1 prior to application on to the target cells. Epithelial cells infected with adenovirus or respiratory syncytial virus showed increased expression of ICAM1; this outcome supported augmented adherence of Tfp-expressing NTHI. Collectively, these data revealed the cognate receptor for NTHI Tfp as ICAM1 and promote continued development of a Tfp-targeted vaccine for NTHI-induced diseases of the airway wherein upper respiratory tract viruses play a key predisposing role.

  16. Cigarette Smoke-Induced Lung Disease Predisposes to More Severe Infection with Nontypeable Haemophilus influenzae: Protective Effects of Andrographolide.

    Science.gov (United States)

    Tan, W S Daniel; Peh, Hong Yong; Liao, Wupeng; Pang, Chu Hui; Chan, Tze Khee; Lau, Suk Hiang; Chow, Vincent T; Wong, W S Fred

    2016-05-27

    Cigarette smoke (CS) is associated with many maladies, one of which is chronic obstructive pulmonary disease (COPD). As the disease progresses, patients are more prone to develop COPD exacerbation episodes by bacterial infection, particularly to nontypeable Haemophilus influenza (NTHi) infection. The present study aimed to develop a CS-exposed mouse model that increases inflammation induced by NTHi challenge and investigate the protective effects of andrographolide, a bioactive molecule with anti-inflammatory and antioxidant properties isolated from the plant Andrographis paniculata. Female BALB/c mice exposed to 2 weeks of CS followed by a single intratracheal instillation of NTHi developed increased macrophage and neutrophil pulmonary infiltration, augmented cytokine levels, and heightened oxidative damage. Andrographolide effectively reduced lung cellular infiltrates and decreased lung levels of TNF-α, IL-1β, CXCL1/KC, 8-OHdG, matrix metalloproteinase-8 (MMP-8), and MMP-9. The protective actions of andrographolide on CS-predisposed NTHi inflammation might be attributable to increased nuclear factor erythroid-2-related factor 2 (Nrf2) activation and decreased Kelch-like ECH-associated protein 1 (Keap1) repressor function, resulting in enhanced gene expression of antioxidant enzymes including heme oxygenase-1 (HO-1), glutathione reductase (GR), glutathione peroxidase-2 (GPx-2), glutamate-cysteine ligase modifier (GCLM), and NAD(P)H quinone oxidoreductase 1 (NQO1). Taken together, these findings strongly support a therapeutic potential for andrographolide in preventing lung inflammation caused by NTHi in cigarette smokers.

  17. A glycoconjugate of Haemophilus influenzae Type b capsular polysaccharide with tetanus toxoid protein: hydrodynamic properties mainly influenced by the carbohydrate.

    Science.gov (United States)

    Abdelhameed, Ali Saber; Adams, Gary G; Morris, Gordon A; Almutairi, Fahad M; Duvivier, Pierre; Conrath, Karel; Harding, Stephen E

    2016-02-26

    Three important physical properties which may affect the performance of glycoconjugate vaccines against serious disease are molar mass (molecular weight), heterogeneity (polydispersity), and conformational flexibility in solution. The dilute solution behaviour of native and activated capsular polyribosylribitol (PRP) polysaccharides extracted from Haemophilus influenzae type b (Hib), and the corresponding glycoconjugate made by conjugating this with the tetanus toxoid (TT) protein have been characterized and compared using a combination of sedimentation equilibrium and sedimentation velocity in the analytical ultracentrifuge with viscometry. The weight average molar mass of the activated material was considerably reduced (Mw ~ 0.24 × 10(6) g.mol(-1)) compared to the native (Mw ~ 1.2 × 10(6) g.mol(-1)). Conjugation with the TT protein yielded large polydisperse structures (of Mw ~ 7.4 × 10(6) g.mol(-1)), but which retained the high degree of flexibility of the native and activated polysaccharide, with frictional ratio, intrinsic viscosity, sedimentation conformation zoning behaviour and persistence length all commensurate with highly flexible coil behaviour and unlike the previously characterised tetanus toxoid protein (slightly extended and hydrodynamically compact structure with an aspect ratio of ~3). This non-protein like behaviour clearly indicates that it is the carbohydrate component which mainly influences the physical behaviour of the glycoconjugate in solution.

  18. Prophage induction in Haemophilus influenzae and its relationship to mutation by chemical and physical agents

    Energy Technology Data Exchange (ETDEWEB)

    Balganesh, M.; Setlow, J.K. (Brookhaven National Lab., Upton, NY (USA))

    1984-01-01

    It is known that UV, X-rays, MMC and MMS are not mutagenic for H. influenzae, whereas HZ, EMS and MNNG are potent mutagens for this bacterium. All of these agents, however, are known to be both mutagenic and able to induce prophage in E. coli. We report here that all the agents except HZ induce prophage in H. influenzae, and EMS even induces in the recombination-defective recl mutant, which is non-inducible by UV, MMC, MNNG and MMS. MMS did not cause single-strand breaks or gaps in DNA synthesized after treatment of H. influenzae, but EMS and MNNG produced them. EMS caused more breaks in DNA synthesized before treatment than in that synthesized after treatment. On the other hand we did observe such breaks or gaps induced in E. coli in DNA synthesized posttreatment by EMS as well as by MMS and MNNG, at comparable survival levels.

  19. Development of a diagnostic real-time polymerase chain reaction assay for the detection of invasive Haemophilus influenzae in clinical samples.

    Science.gov (United States)

    Meyler, Kenneth L; Meehan, Mary; Bennett, Desiree; Cunney, Robert; Cafferkey, Mary

    2012-12-01

    Since the introduction of the Haemophilus influenzae serotype b vaccine, invasive H. influenzae disease has become dominated by nontypeable (NT) strains. Several widely used molecular diagnostic methods have been shown to lack sensitivity or specificity in the detection of some of these strains. Novel real-time assays targeting the fucK, licA, and ompP2 genes were developed and evaluated. The fucK assay detected all strains of H. influenzae tested (n = 116) and had an analytical sensitivity of 10 genome copies/polymerase chain reaction (PCR). This assay detected both serotype b and NT H. influenzae in 12 previously positive specimens (culture and/or bexA PCR) and also detected H. influenzae in a further 5 of 883 culture-negative blood and cerebrospinal fluid (CSF) samples. The fucK assay has excellent potential as a diagnostic test for detection of typeable and nontypeable strains of invasive H. influenzae in clinical samples of blood and CSF. PMID:23017260

  20. Development of a diagnostic real-time polymerase chain reaction assay for the detection of invasive Haemophilus influenzae in clinical samples.

    LENUS (Irish Health Repository)

    Meyler, Kenneth L

    2012-12-01

    Since the introduction of the Haemophilus influenzae serotype b vaccine, invasive H. influenzae disease has become dominated by nontypeable (NT) strains. Several widely used molecular diagnostic methods have been shown to lack sensitivity or specificity in the detection of some of these strains. Novel real-time assays targeting the fucK, licA, and ompP2 genes were developed and evaluated. The fucK assay detected all strains of H. influenzae tested (n = 116) and had an analytical sensitivity of 10 genome copies\\/polymerase chain reaction (PCR). This assay detected both serotype b and NT H. influenzae in 12 previously positive specimens (culture and\\/or bexA PCR) and also detected H. influenzae in a further 5 of 883 culture-negative blood and cerebrospinal fluid (CSF) samples. The fucK assay has excellent potential as a diagnostic test for detection of typeable and nontypeable strains of invasive H. influenzae in clinical samples of blood and CSF.

  1. Prevalence of macrolide-non-susceptible isolates among β-lactamase-negative ampicillin-resistant Haemophilus influenzae in a tertiary care hospital in Japan.

    Science.gov (United States)

    Wajima, Takeaki; Seyama, Shoji; Nakamura, Yuka; Kashima, Chihiro; Nakaminami, Hidemasa; Ushio, Masanobu; Fujii, Takeshi; Noguchi, Norihisa

    2016-09-01

    β-Lactamase-negative ampicillin-resistant (BLNAR) Haemophilus influenzae account for a large portion of H. influenzae clinical isolates in Japan. The aim of this study was to clarify the antimicrobial susceptibility of BLNAR H. influenzae clinical isolates as well as the annual changes in susceptibility. BLNAR H. influenzae isolates were collected from a tertiary care hospital from 2007 to 2012. Antimicrobial susceptibility testing was performed and resistance mechanisms were analysed. All of the isolates (n=304) had amino acid substitutions in penicillin-binding protein 3 (PBP3) and isolates were classified by these amino acid substitutions: R517H or N526K (class I); S385T and R517H (class II); and S385T and N526K (class III). Classes I, II and III represented 8.2% (n=25), 9.5% (n=29) and 81.6% (n=248) of the isolates, respectively; 2 isolates could not be classified because they had a PBP3 with a substantially mutated FtsI transpeptidase domain. All of the isolates were highly susceptible to fluoroquinolones and carbapenems. The number of clarithromycin (CAM)-non-susceptible [minimum inhibitory concentration (MIC) ≥16μg/mL] H. influenzae isolates increased significantly between 2010 and 2012. Moreover, CAM-non-susceptible H. influenzae isolates were prevalent among class II and class III BLNAR H. influenzae. Multilocus sequence typing (MLST) of the CAM-resistant (MIC ≥32μg/mL) H. influenzae isolates showed that they were not specific sequence types, suggesting that CAM resistance may occur in any isolates. These results raise concern regarding the occurrence of multidrug-resistant BLNAR H. influenzae. PMID:27530834

  2. Prevalence of macrolide-non-susceptible isolates among β-lactamase-negative ampicillin-resistant Haemophilus influenzae in a tertiary care hospital in Japan.

    Science.gov (United States)

    Wajima, Takeaki; Seyama, Shoji; Nakamura, Yuka; Kashima, Chihiro; Nakaminami, Hidemasa; Ushio, Masanobu; Fujii, Takeshi; Noguchi, Norihisa

    2016-09-01

    β-Lactamase-negative ampicillin-resistant (BLNAR) Haemophilus influenzae account for a large portion of H. influenzae clinical isolates in Japan. The aim of this study was to clarify the antimicrobial susceptibility of BLNAR H. influenzae clinical isolates as well as the annual changes in susceptibility. BLNAR H. influenzae isolates were collected from a tertiary care hospital from 2007 to 2012. Antimicrobial susceptibility testing was performed and resistance mechanisms were analysed. All of the isolates (n=304) had amino acid substitutions in penicillin-binding protein 3 (PBP3) and isolates were classified by these amino acid substitutions: R517H or N526K (class I); S385T and R517H (class II); and S385T and N526K (class III). Classes I, II and III represented 8.2% (n=25), 9.5% (n=29) and 81.6% (n=248) of the isolates, respectively; 2 isolates could not be classified because they had a PBP3 with a substantially mutated FtsI transpeptidase domain. All of the isolates were highly susceptible to fluoroquinolones and carbapenems. The number of clarithromycin (CAM)-non-susceptible [minimum inhibitory concentration (MIC) ≥16μg/mL] H. influenzae isolates increased significantly between 2010 and 2012. Moreover, CAM-non-susceptible H. influenzae isolates were prevalent among class II and class III BLNAR H. influenzae. Multilocus sequence typing (MLST) of the CAM-resistant (MIC ≥32μg/mL) H. influenzae isolates showed that they were not specific sequence types, suggesting that CAM resistance may occur in any isolates. These results raise concern regarding the occurrence of multidrug-resistant BLNAR H. influenzae.

  3. MICROBIOLOGICAL CHARACTERISATION OF Haemophilus influenzae STRAINS ISOLATED FROM PATIENTS WITH INVASIVE AND RESPIRATORY DISEASES

    Directory of Open Access Journals (Sweden)

    Tomislav Kostyanev

    2010-01-01

    Full Text Available A total of 175 H. influenzae strains were collected between 1994 and 2009 from all aged patient groups. The strains were isolated from patients with invasive and community-acquired respiratory tract infections. All strains were identified according to standard microbiological methods. Serotyping was done by a coagglutination test and by molecular PCR capsular genotyping. Beta-lactamase production was determined by the chromogenic cephalosporin test with nitrocephin as substrate. Most of the isolated H. influenzae strains were from children under 5 years of age (57.7%. Overall, 61 strains belonged to serotype b (34.9% by the means of PCR capsular typing, 1 strain was type f, and 113 isolates (64.6% were non-typeable (non-encapsulated H. influenzae. Among the infants and children with meningitis or other invasive infections, aged 2 month to 5 years, all strains, except one, were serotype b. In respiratory tract infections (pneumonia, otitis media, sinusitis and people with chronic pulmonary diseases - exacerbations of COPD, bronchiectasis, cystic fibrosis the most common - 96.5% were non-typeable strains in both groups children and adults. Overall, the prevalence of beta-lactamase production was 19.4%. But, it was much higher for invasive strains from CSF isolates - 37.7%, 25% in blood samples, and 37.5% in otitis media causative strains. Beta-lactamase production was less frequent in respiratory tract isolates - in sputum 13.3% and in URT samples - 2.3%. The rate of beta-lactamase production in CSF isolates has not changed for the last 10 years.PCR capsular genotyping method has to be performed for all non-b-type strains. The implementation of Hib vaccine in our country will be accompanied by a reduction in invasive diseases caused by H. influenzae type b in children, but it is not useful in preventing infections caused by non-typeable H. influenzae strains.

  4. Haemophilus parasuis exhibits IgA protease activity but lacks homologs of the IgA protease genes of Haemophilus influenzae

    Science.gov (United States)

    Haemophilus parasuis, the bacterium responsible for Glasser's disease, is a pathogen of significant concern in modern high-health swine production systems but there is little information regarding the identity or function of its virulence factors. Several important human mucosal pathogens, including...

  5. Comprehensive Proteomic and Metabolomic Signatures of Nontypeable Haemophilus influenzae-Induced Acute Otitis Media Reveal Bacterial Aerobic Respiration in an Immunosuppressed Environment.

    Science.gov (United States)

    Harrison, Alistair; Dubois, Laura G; St John-Williams, Lisa; Moseley, M Arthur; Hardison, Rachael L; Heimlich, Derek R; Stoddard, Alexander; Kerschner, Joseph E; Justice, Sheryl S; Thompson, J Will; Mason, Kevin M

    2016-03-01

    A thorough understanding of the molecular details of the interactions between bacteria and host are critical to ultimately prevent disease. Recent technological advances allow simultaneous analysis of host and bacterial protein and metabolic profiles from a single small tissue sample to provide insight into pathogenesis. We used the chinchilla model of human otitis media to determine, for the first time, the most expansive delineation of global changes in protein and metabolite profiles during an experimentally induced disease. After 48 h of infection with nontypeable Haemophilus influenzae, middle ear tissue lysates were analyzed by high-resolution quantitative two-dimensional liquid chromatography-tandem mass spectrometry. Dynamic changes in 105 chinchilla proteins and 66 metabolites define the early proteomic and metabolomic signature of otitis media. Our studies indicate that establishment of disease coincides with actin morphogenesis, suppression of inflammatory mediators, and bacterial aerobic respiration. We validated the observed increase in the actin-remodeling complex, Arp2/3, and experimentally showed a role for Arp2/3 in nontypeable Haemophilus influenzae invasion. Direct inhibition of actin branch morphology altered bacterial invasion into host epithelial cells, and is supportive of our efforts to use the information gathered to modify outcomes of disease. The twenty-eight nontypeable Haemophilus influenzae proteins identified participate in carbohydrate and amino acid metabolism, redox homeostasis, and include cell wall-associated metabolic proteins. Quantitative characterization of the molecular signatures of infection will redefine our understanding of host response driven developmental changes during pathogenesis. These data represent the first comprehensive study of host protein and metabolite profiles in vivo in response to infection and show the feasibility of extensive characterization of host protein profiles during disease. Identification of

  6. 流感嗜血杆菌的耐药性及耐药机制%Antimicrobial resistance and resistance mechanism in haemophilus influenzae

    Institute of Scientific and Technical Information of China (English)

    桂和翠; 王中新; 沈纪录

    2011-01-01

    Haemophilus influenzae is the common pathogenic bacteria of respiratory tract infections in human,particularly,in children.Recently,with the widespread use of antibiotics,resistances to various antibiotics such as β-lactams,trimethoprim-sulfamethoxazole,tet-racyclines,chloramphenicol and macrolides become increasingly serious and have distinct differences in different regions.In addition,the growing cases of β-lactmase-negative ampicillin-resistance(BLNAR)show complex resistance mechanism under selecting pressure in haemophilus influenzae.This paper reviews on the antimicrobial resistance and resistance mechanism in haemophilus influenzae.%流感嗜血杆菌是引起人类呼吸道感染的常见致病菌,尤其在儿童中.近年来,由于抗生素的广泛使用,流感嗜血杆菌对β-内酰胺类抗生素、复方新诺明、四环素、氯霉素、大环内酯类的耐药率不断上升,且存在明显的地区差异.此外,β-内酰胺酶阴性的耐氨苄西林(BLNAR)流感嗜血杆菌报道增加,更表明了抗生素选择压力下复杂的耐药机制.该文就流感嗜血杆菌对常用抗生素的耐药性和耐药机制作一综述.

  7. NMR structure of HI0004, a putative essential gene product from Haemophilus influenzae, and comparison with the X-ray structure of an Aquifex aeolicus homolog

    OpenAIRE

    Yeh, Deok Cheon; Lisa M Parsons; Parsons, James F.; Liu, Fang; Eisenstein, Edward; Orban, John

    2005-01-01

    The solution structure of the 154-residue conserved hypothetical protein HI0004 has been determined using multidimensional heteronuclear NMR spectroscopy. HI0004 has sequence homologs in many organisms ranging from bacteria to humans and is believed to be essential in Haemophilus influenzae, although an exact function has yet to be defined. It has a α–β–α sandwich architecture consisting of a central four-stranded β-sheet with the α2-helix packed against one side of the β-sheet and four α-hel...

  8. Elevated levels of maternal anti-tetanus toxin antibodies do not suppress the immune response to a Haemophilus influenzae type b polyribosylphosphate-tetanus toxoid conjugate vaccine.

    OpenAIRE

    Panpitpat, C.; Thisyakorn, U.; Chotpitayasunondh, T; Fürer, E; Que, J U; Hasler, T.; Cryz, S J

    2000-01-01

    Reported are the effects of elevated levels of anti-tetanus antibodies on the safety and immune response to a Haemophilus influenzae type b polyribosylphosphate (PRP)-tetanus toxoid conjugate (PRP-T) vaccine. A group of Thai infants (n = 177) born to women immunized against tetanus during pregnancy were vaccinated with either a combined diphtheria-tetanus-pertussis (DTP) PRP-T vaccine or DTP and a PRP-conjugate vaccine using Neisseria meningitidis group B outer-membrane proteins as a carrier ...

  9. The Haemophilus influenzae dprABC genes constitute a competence-inducible operon that requires the product of the tfoX (sxy) gene for transcriptional activation.

    OpenAIRE

    Karudapuram, S; Barcak, G J

    1997-01-01

    We previously showed that dprA is required for efficient processing of linear DNA during cellular transformation in Haemophilus influenzae. In this study the transcriptional regulation of dprA and two downstream genes, dprB and dprC, is examined. We demonstrate by Northern blot analysis that the dprABC genes are transcriptionally coregulated and competence inducible. We used primer extension analysis to map the transcriptional start site of dprA and of rec-2, another transformation gene invol...

  10. Impaired Haemophilus influenzae Type b Transplacental Antibody Transmission and Declining Antibody Avidity through the First Year of Life Represent Potential Vulnerabilities for HIV-Exposed but -Uninfected Infants

    OpenAIRE

    Gaensbauer, James T.; Rakhola, Jeremy T.; Onyango-Makumbi, Carolyne; Mubiru, Michael; Westcott, Jamie E; Krebs, Nancy F; Asturias, Edwin J; Fowler, Mary Glenn; McFarland, Elizabeth; Janoff, Edward N.

    2014-01-01

    To determine whether immune function is impaired among HIV-exposed but -uninfected (HEU) infants born to HIV-infected mothers and to identify potential vulnerabilities to vaccine-preventable infection, we characterized the mother-to-infant placental transfer of Haemophilus influenzae type b-specific IgG (Hib-IgG) and its levels and avidity after vaccination in Ugandan HEU infants and in HIV-unexposed U.S. infants. Hib-IgG was measured by enzyme-linked immunosorbent assay in 57 Ugandan HIV-inf...

  11. Heavy-chain isotype patterns of human antibody-secreting cells induced by Haemophilus influenzae type b conjugate vaccines in relation to age and preimmunity

    DEFF Research Database (Denmark)

    Barington, T; Juul, Lars; Gyhrs, A;

    1994-01-01

    The influence of preexisting immunity on the heavy-chain isotypes of circulating antibody-secreting cells (AbSC) induced by vaccination with Haemophilus influenzae type b (Hib) capsular polysaccharide (HibCP) coupled to tetanus toxoid (TT) or diphtheria toxoid (DT) and by vaccination with TT or DT...... of natural HibCP antibodies (r = 0.59; P = 0.00002). A possible role of natural exposure for Hib or cross-reactive bacteria on the mucosal surfaces in the shaping of the isotype response to HibCP conjugate vaccines is discussed....

  12. Quantitative detection of Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae in patients with new influenza A (H1N1/2009 and influenza A/2010 virus infection

    Directory of Open Access Journals (Sweden)

    Safaeyan, Firouzeh

    2015-04-01

    Full Text Available Introduction: Viral influenza is a seasonal infection associated with significant morbidity and mortality. In the United States more than 35,000 deaths and 200,000 hospitalizations are recorded annually due to influenza. Secondary bacterial infections or co-infections associated with cases of influenza are a leading cause of severe morbidity and mortality, especially among high-risk groups such as the elderly and young children. Aim: The aim of the present study was the quantitative detection of and in a group of patients with seasonal influenza A, influenza A ( pandemic 2009, and patients with symptoms of respiratory infection, but the negative for serving as control group.Method: In total, 625 patients suspected respiratory infection from April 2009 to April 2010 were studied. There were 58 patients with influenza A and 567 patients negative for influenza A . From November 2010 to February 2011, 158 patients with respiratory symptoms were analyzed for seasonal influenza A. There were 25 patients with seasonal influenza A. To check the colonization status among the healthy individuals 62 healthy persons were further investigated. Individual were screened in parallel. The choices of special genes were amplified from clinical specimens using real-time PCR with a cutoff of 10 CFU/mL to differentiate colonization from infection in respiratory tract.Results: and were detected in 12%, 26% and 33% of patients with , while the corresponding figures were 9%, 19%, and 31% for negative patients. Among patients with seasonal influenza A 12% 24% , and 32% co-infections were detected, while influenza negative control group yielded 5% , 11% , and 10% , respectively. Conclusion: The results of this study indicated that the serotype of pandemic 2009 did not increase incidence of secondary infection with and . Quantitative detection of secondary bacterial infection by QR-PCR can help us for distinguishing colonization from infection and controlling misuse of

  13. [Comparison of culture and real-time PCR methods in the detection of Streptococcus pneumoniae and Haemophilus influenzae in acute otitis media effusion specimens].

    Science.gov (United States)

    Eser, Ozgen Köseoğlu; Alp, Sehnaz; Ergin, Alper; Ipçi, Kaan; Alp, Alpaslan; Gür, Deniz; Hasçelik, Gülşen

    2012-10-01

    Streptococcus pneumoniae and Haemophilus influenzae are the major etiologic agents of acute otitis media. This study was aimed to compare the detection rate of S.pneumoniae and H.influenzae by culture and real-time polymerase chain reaction (Rt-PCR) in the middle ear effusions of patients diagnosed as acute otitis media. A total of 60 middle ear effusion samples collected from children with acute otitis media were included in the study. The samples were inoculated and incubated in BACTEC Ped Plus blood culture bottles and BACTEC 9120 system (BD Diagnostic Systems, MD), respectively, and the isolates were identified by conventional methods. For the molecular diagnosis of H.influenzae and S.pneumoniae, ply pneumolysin gene and HIB capsule region, respectively were amplified by Rt-PCR (LightCycler, Roche Diagnostics, Germany). H.influenzae and S.pneumoniae were isolated from 5 (8.3%) and 3 (5%) of the patient samples with conventional culture methods, respectively. In addition in 11.6% of the samples other microorganisms (Staphylococcus epidermidis, Streptococcus intermedius, Streptococcus sanguinis, Moraxella catarrhalis, Pseudomonas aeruginosa, Candida albicans) were also isolated. On the other hand H.influenzae and S.pneumoniae were detected in 38 (63.3%) and 24 (40%) of the samples with Rt-PCR, respectively. There was about eight fold increase in the detection frequency of H.influenzae and S.pneumoniae with Rt-PCR compared to culture methods. When culture was accepted as the gold standard method, the sensitivity, specificity and positive predictive value of Rt-PCR in the detection of H.influenzae and S.pneumoniae were estimated as 80%, 51% and 98.2%, respectively. As a result, Rt-PCR was shown to be a sensitive method and could be preferred for the rapid diagnosis of H.influenzae and S.pneumoniae in the etiological diagnosis of acute otitis media, especially in culture negative cases.

  14. Infant rat infection modifies phenotypic properties of an invasive nontypeable Haemophilus influenzae

    OpenAIRE

    Tsao, David; Nelson, Kevin L.; Kim, Daniel; Smith, Arnold L.

    2011-01-01

    Enhancing the virulence trait of a specific bacterium in an animal model is often performed prior to the use of the strain for ex vivo human studies, such as reactivity with complement and antibody, or with phagocytic cells. For example, in S. penumoniae mouse passage is used to enhance capsule production. While investigating an unusual serum-resistant unencapsulated H. influenzae (R2866), we found that animal passage yielded an isolate (R3392) which had decreased resistance to human serum, b...

  15. Non-typeable Haemophilus influenzae and Streptococcus pneumoniae as primary causes of acute otitis media in colombian children: a prospective study

    Directory of Open Access Journals (Sweden)

    Castrejon Maria M

    2011-01-01

    Full Text Available Abstract Background Acute otitis media (AOM is one of the most frequently encountered bacterial infections in children aged Streptococcus pneumoniae (S. pneumoniae and non-typeable Haemophilus influenzae (NTHi are historically identified as primary AOM causes. Nevertheless, recent data on bacterial pathogens causing AOM in Latin America are limited. This prospective study aimed to identify and characterize bacterial etiology and serotypes of AOM cases including antimicrobial susceptibility in Methods From February 2008 to January 2009, children ≥3 months and Results Of the 106 enrolled children, 99 were included in the analysis. Bacteria were cultured from 62/99 (63% of samples with S. pneumoniae, H. influenzae, or S. pyogenes. The most commonly isolated bacteria were H. influenzae in 31/99 (31% and S. pneumoniae in 30/99 (30% of samples. The majority of H. influenzae episodes were NTHi (27/31; 87%. 19F was the most frequently isolated pneumococcal serotype (10/30; 33%. Of the 30 S. pneumoniae positive samples, 8/30 (27% were resistant to tetracycline, 5/30 (17% to erythromycin and 8/30 (27% had intermediate resistance to penicillin. All H. influenzae isolates tested were negative to beta-lactamase. Conclusions NTHi and S. pneumoniae are the leading causes of AOM in Colombian children. A pneumococcal conjugate vaccine that prevents both pathogens could be useful in maximizing protection against AOM.

  16. Immune responses to HBsAg conjugated to protein D of non-typeable Haemophilus influenzae in mice.

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    Qiudong Su

    Full Text Available Hepatitis B vaccine that contains an aluminum hydroxide adjuvant induces apoptotic death of Hepa 1-6 cells. Difficult-to-degrade chemical additives in vaccines effectively enhance vaccine immunogenicity, but also affect the host tissue. Identification of bio-molecules that are readily degraded and compatible in vivo as an adjuvant is important for vaccine research. The hapten-carrier effect suggests that stimulation of helper T (Th cells by carrier adjuvants is feasible. Protein D (PD of non-typeable Haemophilus influenzae covalently conjugated to some polysaccharide vaccines has been confirmed to convert T-cell independent (TI antigens into T-cell dependent (TD antigens, and elicit strong T-cell responses ultimately. Herein, we would substitube PD for aluminum hydroxide adjuvant in Hepatitis B vaccine.Truncated PD (amino acids 20-364 was expressed in Escherichia coli and purified by (NH42SO4 precipitation and DEAE chromatography. After evaluation of antigenicity by western blotting, PD was covalently conjugated to yeast-derived recombinant HBsAg by cross-linking with glutaraldehyde. Intramuscular immunization with the conjugate induced higher level of HBsAg-specific antibody than did HBsAg alone (p < 0.05, and was comparable to commercial Hepatitis B vaccine. During the surveillance period (days 35-105, anti-HBs titers were hold high. Moreover, the conjugated vaccine enhanced Th1 immune responses, while Th2 responses were also activated and induced an antibody response, as determined by IFN-γ ELISPOT and IgG1/IgG2a ratio assays.Recombinant truncated PD covalently conjugated to HBsAg antigen enhanced the immunogenicity of the antigen in mice simultaneously by humoral and cellular immune response, which would facilitate therapeutic hepatitis B vaccines.

  17. A new model for non-typeable Haemophilus influenzae middle ear infection in the Junbo mutant mouse

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    Derek Hood

    2016-01-01

    Full Text Available Acute otitis media, inflammation of the middle ear, is the most common bacterial infection in children and, as a consequence, is the most common reason for antimicrobial prescription to this age group. There is currently no effective vaccine for the principal pathogen involved, non-typeable Haemophilus influenzae (NTHi. The most frequently used and widely accepted experimental animal model of middle ear infection is in chinchillas, but mice and gerbils have also been used. We have established a robust model of middle ear infection by NTHi in the Junbo mouse, a mutant mouse line that spontaneously develops chronic middle ear inflammation in specific pathogen-free conditions. The heterozygote Junbo mouse (Jbo/+ bears a mutation in a gene (Evi1, also known as Mecom that plays a role in host innate immune regulation; pre-existing middle ear inflammation promotes NTHi middle ear infection. A single intranasal inoculation with NTHi produces high rates (up to 90% of middle ear infection and bacterial titres (104-105 colony-forming units/µl in bulla fluids. Bacteria are cleared from the majority of middle ears between day 21 and 35 post-inoculation but remain in approximately 20% of middle ears at least up to day 56 post-infection. The expression of Toll-like receptor-dependent response cytokine genes is elevated in the middle ear of the Jbo/+ mouse following NTHi infection. The translational potential of the Junbo model for studying antimicrobial intervention regimens was shown using a 3 day course of azithromycin to clear NTHi infection, and its potential use in vaccine development studies was shown by demonstrating protection in mice immunized with killed homologous, but not heterologous, NTHi bacteria.

  18. Evaluation of Haemophilus influenzae type b carrier status among children 10 years after the introduction of Hib vaccine in Brazil.

    Science.gov (United States)

    Zanella, Rosemeire Cobo; Brandileone, Maria Cristina de Cunto; Andrade, Ana Lúcia; Ogassavara, Cinthya Terumi; Fiório, Cleiton Eduardo; Brandão, Angela Pires; Almeida, Samanta Cristine Grassi; Lemos, Ana Paula Silva; Gorla, Maria Cecília; Carvalhanas, Telma Regina; Sato, Helena; Liphaus, Bernadete; Nerger, Maria Lígia; Conde, Monica; Ribeiro, Ana Freitas

    2015-09-01

    The aim of the present study was to assess the prevalence of Haemophilus influenzae type b (Hib) nasopharyngeal (NP) colonisation among healthy children where Hib vaccination using a 3p+0 dosing schedule has been routinely administered for 10 years with sustained coverage (> 90%). NP swabs were collected from 2,558 children who had received the Hib vaccine, of whom 1,379 were 12-< 24 months (m) old and 1,179 were 48-< 60 m old. Hi strains were identified by molecular methods. Hi carriage prevalence was 45.1% (1,153/2,558) and the prevalence in the 12-< 24 m and 48-< 60 m age groups were 37.5% (517/1,379) and 53.9% (636/1,179), respectively. Hib was identified in 0.6% (16/2,558) of all children in the study, being 0.8% (11/1,379) and 0.4% (5/1,179) among the 12-< 24 m and 48-< 60 m age groups, respectively. The nonencapsulate Hi colonisation was 43% (n = 1,099) and was significantly more frequent at 48-< 60 m of age (51.6%, n = 608) compared with that at 12-< 24 m of age (35.6%, n = 491). The overall resistance rates to ampicillin and chloramphenicol were 16.5% and 3.7%, respectively; the co-resistance was detected in 2.6%. Our findings showed that the Hib carrier rate in healthy children under five years was very low after 10 years of the introduction of the Hib vaccine. PMID:26517654

  19. Evaluation of Haemophilus influenzae type b carrier status among children 10 years after the introduction of Hib vaccine in Brazil

    Science.gov (United States)

    Zanella, Rosemeire Cobo; Brandileone, Maria Cristina de Cunto; Andrade, Ana Lúcia; Ogassavara, Cinthya Terumi; Fiório, Cleiton Eduardo; Brandão, Angela Pires; Almeida, Samanta Cristine Grassi; Lemos, Ana Paula Silva; Gorla, Maria Cecília; Carvalhanas, Telma Regina; Sato, Helena; Liphaus, Bernadete; Nerger, Maria Lígia; Conde, Monica; Ribeiro, Ana Freitas

    2015-01-01

    The aim of the present study was to assess the prevalence of Haemophilus influenzae type b (Hib) nasopharyngeal (NP) colonisation among healthy children where Hib vaccination using a 3p+0 dosing schedule has been routinely administered for 10 years with sustained coverage (> 90%). NP swabs were collected from 2,558 children who had received the Hib vaccine, of whom 1,379 were 12-< 24 months (m) old and 1,179 were 48-< 60 m old. Hi strains were identified by molecular methods. Hi carriage prevalence was 45.1% (1,153/2,558) and the prevalence in the 12-< 24 m and 48-< 60 m age groups were 37.5% (517/1,379) and 53.9% (636/1,179), respectively. Hib was identified in 0.6% (16/2,558) of all children in the study, being 0.8% (11/1,379) and 0.4% (5/1,179) among the 12-< 24 m and 48-< 60 m age groups, respectively. The nonencapsulate Hi colonisation was 43% (n = 1,099) and was significantly more frequent at 48-< 60 m of age (51.6%, n = 608) compared with that at 12-< 24 m of age (35.6%, n = 491). The overall resistance rates to ampicillin and chloramphenicol were 16.5% and 3.7%, respectively; the co-resistance was detected in 2.6%. Our findings showed that the Hib carrier rate in healthy children under five years was very low after 10 years of the introduction of the Hib vaccine. PMID:26517654

  20. Invasive Haemophilus influenzae infections in Germany: impact of non-type b serotypes in the post-vaccine era

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    Milde-Busch Astrid

    2009-04-01

    Full Text Available Abstract Background Haemophilus influenzae type b (Hib vaccination led to a significant decrease in invasive bacterial infections in children. The aim of this study was to assess a potential shift to more non-type b invasive infections in a population with high Hib vaccination coverage and to compare the burden of suffering between children with Hib, capsulated non-b and non-capsulated Hi infections. Methods Cases with confirmed invasive Hi infections were ascertained through two independent nationwide active surveillance systems in 1998–2005. Information on possible predisposing conditions and clinical information was available from 2001 onwards. Results The total number of reported non-type b Hi cases varied between 10 cases in 1998, 27 in 2000 and 14 in 2005. In each year, non-capsulated serotypes outnumbered capsulated non-type b ones. 192 cases were detected in 2001–2005, more than one half was non-type b and 88% of the non-type b cases were non-capsulated. For cases with Hib/capsulated non-type b infections the most common clinical presentation was meningitis (67% each; 89%/78% had no potential predisposing condition, 75%/72% completely recovered from disease and 6% (each died. In contrast, meningitis was diagnosed in 34% of the non-capsulated Hi infections, septicaemia in 28% and pneumonia 21%; 62% had no potential predisposing condition, 83% completely recovered and 3% died. Conclusion There was no increase in non-type b Hi invasive infections during 8 years of active surveillance in Germany. Invasive disease due to non-type b Hi is not confined to children with risk factors. In patients with capsulated non-type b Hi infections the proportion of meningitis cases is similar to Hib, but double as high as in non-capsulated Hi.

  1. Beta- lactam antibiotics stimulate biofilm formation in non-typeable haemophilus influenzae by up-regulating carbohydrate metabolism.

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    Siva Wu

    Full Text Available Non-typeable Haemophilus influenzae (NTHi is a common acute otitis media pathogen, with an incidence that is increased by previous antibiotic treatment. NTHi is also an emerging causative agent of other chronic infections in humans, some linked to morbidity, and all of which impose substantial treatment costs. In this study we explore the possibility that antibiotic exposure may stimulate biofilm formation by NTHi bacteria. We discovered that sub-inhibitory concentrations of beta-lactam antibiotic (i.e., amounts that partially inhibit bacterial growth stimulated the biofilm-forming ability of NTHi strains, an effect that was strain and antibiotic dependent. When exposed to sub-inhibitory concentrations of beta-lactam antibiotics NTHi strains produced tightly packed biofilms with decreased numbers of culturable bacteria but increased biomass. The ratio of protein per unit weight of biofilm decreased as a result of antibiotic exposure. Antibiotic-stimulated biofilms had altered ultrastructure, and genes involved in glycogen production and transporter function were up regulated in response to antibiotic exposure. Down-regulated genes were linked to multiple metabolic processes but not those involved in stress response. Antibiotic-stimulated biofilm bacteria were more resistant to a lethal dose (10 µg/mL of cefuroxime. Our results suggest that beta-lactam antibiotic exposure may act as a signaling molecule that promotes transformation into the biofilm phenotype. Loss of viable bacteria, increase in biofilm biomass and decreased protein production coupled with a concomitant up-regulation of genes involved with glycogen production might result in a biofilm of sessile, metabolically inactive bacteria sustained by stored glycogen. These biofilms may protect surviving bacteria from subsequent antibiotic challenges, and act as a reservoir of viable bacteria once antibiotic exposure has ended.

  2. Immunogenicity, reactogenicity and consistency of production of a Brazilian combined vaccine against diphtheria, tetanus, pertussis and Haemophilus influenzae type b

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    Reinaldo de Menezes Martins

    2008-11-01

    Full Text Available A randomized, double-blinded study evaluating the immunogenicity, safety and consistency of production of a combined diphtheria-tetanus-pertussis-Haemophilus influenzae type b vaccine entirely produced in Brazil by Bio-Manguinhos and Instituto Butantan (DTP/Hib-BM was undertaken. The reference vaccine had the same DTP vaccine but the Hib component was produced using purified materials supplied by GlaxoSmithKline (DTP/Hib-GSK, which is registered and has supplied the Brazilian National Immunization Program for over more than five years. One thousand infants were recruited for the study and received vaccinations at two, four and six months of age. With respect to immunogenicity, the vaccination protocol was followed in 95.6% and 98.4% of infants in the DTP/Hib-BM and DTP/Hib-GSK groups, respectively. For the Hib component of the study, there was 100% seroprotection (>0.15 µg/mL with all three lots of DTP/Hib-BM and DTP/Hib-GSK. The geometric mean titer (GMT was 9.3 µg/mL, 10.3 µg/mL and 10.3 µg/mL for lots 1, 2 and 3 of DTP/Hib-BM, respectively, and the GMT was 11.3 g/mL for DTP/Hib-GSK. For diphtheria, tetanus and pertussis, seroprotection was 99.7%, 100% and 99.9%, respectively, for DTP/Hib-BM, three lots altogether and 99.2%, 100% and 100% for DTP/Hib-GSK. GMTs were similar across all lots and vaccines. Adverse events rates were comparable among the vaccine groups. The Brazilian DTP/Hib vaccine demonstrated an immunogenicity and reactogenicity profile similar to that of the reference vaccine.

  3. Dps promotes survival of nontypeable Haemophilus influenzae in biofilm communities in vitro and resistance to clearance in vivo

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    Bing ePang

    2012-05-01

    Full Text Available Nontypeable Haemophilus influenzae (NTHi is a common airway commensal and opportunistic pathogen that persists within surface-attached biofilm communities. In this study, we tested the hypothesis that bacterial stress-responses are activated within biofilms. Transcripts for several factors associated with bacterial resistance to environmental stress were increased in biofilm cultures as compared to planktonic cultures. Among these, a homolog of the DNA-binding protein from starved cells (dps was chosen for further study. An isogenic NTHi 86-028NP dps mutant was generated and tested for resistance to environmental stress, revealing a significant survival defects in high-iron conditions, which was mediated by oxidative stress and was restored by genetic complementation. As expected, NTHi 86-028NP dps had a general stress-response defect, exhibiting decreased resistance to many types of environmental stress. While no differences were observed in density and structure of NTHi 86-028NP and NTHi 86-028NP dps biofilms, bacterial survival was decreased in NTHi 86-028NP dps biofilms as compared to the parental strain. The role of dps persistence in vivo was tested in animal infection studies. NTHi 86-028NP dps had decreased resistance to clearance after pulmonary infection of elastase-treated mice as compared to NTHi 86-028NP, whereas minimal differences were observed in clearance from mock-treated mice. Similarly, lower numbers of NTHi 86-028NP dps were recovered from middle-ear effusions and bullar homogenates in the chinchilla model for otitis media. Therefore, we conclude that Dps promotes bacterial survival within NTHi biofilm communities both in vitro and in chronic infections in vivo.

  4. The modA10 phasevarion of nontypeable Haemophilus influenzae R2866 regulates multiple virulence-associated traits.

    Science.gov (United States)

    VanWagoner, Timothy M; Atack, John M; Nelson, Kevin L; Smith, Hannah K; Fox, Kate L; Jennings, Michael P; Stull, Terrence L; Smith, Arnold L

    2016-03-01

    Non-typeable Haemophilus influenzae (NTHi) is a human restricted commensal and pathogen that elicits inflammation by adhering to and invading airway epithelia cells: transcytosis across these cells can result in systemic infection. NTHi strain R2866 was isolated from the blood of a normal 30-month old infant with meningitis, and is unusual for NTHi in that it is able to cause systemic infection. Strain R2866 is able to replicate in normal human serum due to expression of lgtC which mimics human blood group p(k). R2866 contains a phase-variable DNA methyltransferase, modA10 which switches ON and OFF randomly and reversibly due to polymerase slippage over a long tetrameric repeat tract located in its open reading frame. Random gain or loss of repeats during replication can results in expressed (ON), or not expressed (OFF) states, the latter due to a frameshift or transcriptional termination at a premature stop codon. We sought to determine if the unusual virulence of R2866 was modified by modA10 phase-variation. A modA10 knockout mutant was found to have increased adherence to, and invasion of, human ear and airway monolayers in culture, and increased invasion and transcytosis of polarized human bronchial epithelial cells. Intriguingly, the rate of bacteremia was lower in the infant rat model of infection than a wild-type R2866 strain, but the fatality rate was greater. Transcriptional analysis comparing the modA10 knockout to the R2866 wild-type parent strain showed increased expression of genes in the modA10 knockout whose products mediate cellular adherence. We conclude that loss of ModA10 function in strain R2866 enhances colonization and invasion by increasing expression of genes that allow for increased adherence, which can contribute to the increased virulence of this strain.

  5. LytM Proteins Play a Crucial Role in Cell Separation, Outer Membrane Composition, and Pathogenesis in Nontypeable Haemophilus influenzae

    Science.gov (United States)

    Ercoli, Giuseppe; Tani, Chiara; Pezzicoli, Alfredo; Vacca, Irene; Martinelli, Manuele; Pecetta, Simone; Petracca, Roberto; Rappuoli, Rino; Pizza, Mariagrazia; Soriani, Marco

    2015-01-01

    ABSTRACT LytM proteins belong to a family of bacterial metalloproteases. In Gram-negative bacteria, LytM factors are mainly reported to have a direct effect on cell division by influencing cleavage and remodeling of peptidoglycan. In this study, mining nontypeable Haemophilus influenzae (NTHI) genomes, three highly conserved open reading frames (ORFs) containing a LytM domain were identified, and the proteins encoded by the ORFs were named YebA, EnvC, and NlpD on the basis of their homology with the Escherichia coli proteins. Immunoblotting and confocal analysis showed that while NTHI NlpD is exposed on the bacterial surface, YebA and EnvC reside in the periplasm. NTHI ΔyebA and ΔnlpD deletion mutants revealed an aberrant division phenotype characterized by an altered cell architecture and extensive membrane blebbing. The morphology of the ΔenvC deletion mutant was identical to that of the wild-type strain, but it showed a drastic reduction of periplasmic proteins, including the chaperones HtrA, SurA, and Skp, and an accumulation of β-barrel-containing outer membrane proteins comprising the autotransporters Hap, IgA serine protease, and HMW2A, as observed by proteomic analysis. These data suggest that EnvC may influence the bacterial surface protein repertoire by facilitating the passage of the periplasmic chaperones through the peptidoglycan layer to the close vicinity of the inner face of the outer membrane. This hypothesis was further corroborated by the fact that an NTHI envC defective strain had an impaired capacity to adhere to epithelial cells and to form biofilm. Notably, this strain also showed a reduced serum resistance. These results suggest that LytM factors are not only important components of cell division but they may also influence NTHI physiology and pathogenesis by affecting membrane composition. PMID:25714719

  6. Structure of YciA from Haemophilus influenzae (HI0827), a Hexameric Broad Specificity Acyl-Coenzyme A Thioesterase

    Energy Technology Data Exchange (ETDEWEB)

    Willis, Mark A.; Zhuang, Zhihao; Song, Feng; Howard, Andrew; Dunaway-Mariano, Debra; Herzberg, Osnat (UNM); (IIT); (UMBI)

    2008-04-02

    The crystal structure of HI0827 from Haemophilus influenzae Rd KW20, initially annotated 'hypothetical protein' in sequence databases, exhibits an acyl-coenzyme A (acyl-CoA) thioesterase 'hot dog' fold with a trimer of dimers oligomeric association, a novel assembly for this enzyme family. In studies described in the preceding paper [Zhuang, Z., Song, F., Zhao, H., Li, L., Cao, J., Eisenstein, E., Herzberg, O., and Dunaway-Mariano, D. (2008) Biochemistry 47, 2789-2796], HI0827 is shown to be an acyl-CoA thioesterase that acts on a wide range of acyl-CoA compounds. Two substrate binding sites are located across the dimer interface. The binding sites are occupied by two CoA molecules, one with full occupancy and the second only partially occupied. The CoA molecules, acquired from HI0827-expressing Escherichia coli cells, remained tightly bound to the enzyme through the protein purification steps. The difference in CoA occupancies indicates a different substrate affinity for each of the binding sites, which in turn implies that the enzyme might be subject to allosteric regulation. Mutagenesis studies have shown that the replacement of the putative catalytic carboxylate Asp44 with an alanine residue abolishes activity. The impact of this mutation is seen in the crystal structure of D44A HI0827. Whereas the overall fold and assembly of the mutant protein are the same as those of the wild-type enzyme, the CoA ligands are absent. The dimer interface is perturbed, and the channel that accommodates the thioester acyl chain is more open and wider than that observed in the wild-type enzyme. A model of intact substrate bound to wild-type HI0827 provides a structural rationale for the broad substrate range.

  7. A new model for non-typeable Haemophilus influenzae middle ear infection in the Junbo mutant mouse

    Science.gov (United States)

    Hood, Derek; Moxon, Richard; Purnell, Tom; Richter, Caroline; Williams, Debbie; Azar, Ali; Crompton, Michael; Wells, Sara; Fray, Martin; Brown, Steve D. M.; Cheeseman, Michael T.

    2016-01-01

    ABSTRACT Acute otitis media, inflammation of the middle ear, is the most common bacterial infection in children and, as a consequence, is the most common reason for antimicrobial prescription to this age group. There is currently no effective vaccine for the principal pathogen involved, non-typeable Haemophilus influenzae (NTHi). The most frequently used and widely accepted experimental animal model of middle ear infection is in chinchillas, but mice and gerbils have also been used. We have established a robust model of middle ear infection by NTHi in the Junbo mouse, a mutant mouse line that spontaneously develops chronic middle ear inflammation in specific pathogen-free conditions. The heterozygote Junbo mouse (Jbo/+) bears a mutation in a gene (Evi1, also known as Mecom) that plays a role in host innate immune regulation; pre-existing middle ear inflammation promotes NTHi middle ear infection. A single intranasal inoculation with NTHi produces high rates (up to 90%) of middle ear infection and bacterial titres (104-105 colony-forming units/µl) in bulla fluids. Bacteria are cleared from the majority of middle ears between day 21 and 35 post-inoculation but remain in approximately 20% of middle ears at least up to day 56 post-infection. The expression of Toll-like receptor-dependent response cytokine genes is elevated in the middle ear of the Jbo/+ mouse following NTHi infection. The translational potential of the Junbo model for studying antimicrobial intervention regimens was shown using a 3 day course of azithromycin to clear NTHi infection, and its potential use in vaccine development studies was shown by demonstrating protection in mice immunized with killed homologous, but not heterologous, NTHi bacteria. PMID:26611891

  8. Sialic acid transport and catabolism are cooperatively regulated by SiaR and CRP in nontypeable Haemophilus influenzae

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    Johnston Jason W

    2010-09-01

    Full Text Available Abstract Background The transport and catabolism of sialic acid, a critical virulence factor for nontypeable Haemophilus influenzae, is regulated by two transcription factors, SiaR and CRP. Results Using a mutagenesis approach, glucosamine-6-phosphate (GlcN-6P was identified as a co-activator for SiaR. Evidence for the cooperative regulation of both the sialic acid catabolic and transport operons suggested that cooperativity between SiaR and CRP is required for regulation. cAMP was unable to influence the expression of the catabolic operon in the absence of SiaR but was able to induce catabolic operon expression when both SiaR and GlcN-6P were present. Alteration of helical phasing supported this observation by uncoupling SiaR and CRP regulation. The insertion of one half-turn of DNA between the SiaR and CRP operators resulted in the loss of SiaR-mediated repression of the transport operon while eliminating cAMP-dependent induction of the catabolic operon when GlcN-6P was present. SiaR and CRP were found to bind to their respective operators simultaneously and GlcN-6P altered the interaction of SiaR with its operator. Conclusions These results suggest multiple novel features for the regulation of these two adjacent operons. SiaR functions as both a repressor and an activator and SiaR and CRP interact to regulate both operons from a single set of operators.

  9. Antimicrobial resistance in Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and group A beta-haemolytic streptococci in 2002-2003. Results of the multinational GRASP Surveillance Program

    DEFF Research Database (Denmark)

    Beekmann, Susan E; Heilmann, Kris P; Richter, Sandra S;

    2005-01-01

    A multinational surveillance study, GRASP, was conducted between November 2002 and April 2003 with the aim of assessing rates of antimicrobial resistance among 2656 isolates of Streptococcus pneumoniae, 2486 isolates of group A beta-haemolytic streptococci, 1358 isolates of Haemophilus influenzae...... and 1047 of Moraxella catarrhalis from 20 countries in Europe, eastern Asia and southern Africa. Conspicuous differences between various countries were noted in the S. pneumoniae resistance rates observed for penicillin (0-79.2%) and erythromycin (4-66%), along with other antimicrobials. The percentage...... of MDR strains was above 25% in 8 of the 20 countries studied. Group A streptococcal macrolide resistance rates ranged from 0% to 35% by country, while rates of beta-lactamase production ranged from 0% to 39% for H. influenzae and 80-100% for M. catarrhalis. Antibiotic resistance in S. pneumoniae remains...

  10. VERIFICATION OF THE PRESENCE OF CAPSULE GENE SEQUENCES IN NASOPHARYNGEAL ISOLATES OF NONTYPEABLE HAEMOPHILUS INFLUENZAE FROM HEALTHY CHILDREN AT A BRAZILIAN DAY CARE CENTER Verificação da presença de seqüências do gene da cápsula em cepas não tipáveis de Haemophilus influenzae isoladas da nasofaringe de crianças saudáveis em uma creche brasileira

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    Maria Emilia Bonifácio da Silva

    2001-10-01

    Full Text Available Fifty-eight nasopharyngeal isolates of Haemophilus influenzae were collected from healthy children at a day care center, and nontypeable isolates were examined by Southern blot for the presence of capsule gene sequences. Seven isolates (12% demonstrated homology with capsule-specific sequences. One isolate was characterized as an H. influenzae type b capsule-deficient strain.Cinqüenta e oito cepas de Haemophilus influenzae foram isoladas da nasofaringe de crianças saudáveis que freqüentam uma creche, e através da técnica de Southern blot foi pesquisada nas cepas acapsuladas a presença de seqüências do gene capsular. Sete cepas (12% caracterizadas sorologicamente como acapsuladas mostraram homologia com seqüências específicas da cápsula. Uma cepa foi caracterizada com uma linhagem H. influenzae tipo b cápsula deficiente.

  11. Recombinant C-terminal 311 amino acids of HapS adhesin as a vaccine candidate for nontypeable Haemophilus influenzae: A study on immunoreactivity in Balb/C mouse.

    Science.gov (United States)

    Tabatabaee Bafroee, Akram Sadat; Siadat, Seyed Davar; Mousavi, Seyed Fazlollah; Aghasadeghi, Mohammad Reza; Khorsand, Hashem; Nejati, Mehdi; Sadat, Seyed Mehdi; Mahdavi, Mehdi

    2016-09-01

    Hap, an auto-transporter protein, is an antigenically conserved adhesion protein which is present on both typeable and nontypeable Haemophilus influenzae. This protein has central role in bacterial attachment to respiratory tract epithelial cells. A 1000bp C-terminal fragment of Hap passenger domain (HapS) from nontypeable Haemophilus influenzae was cloned into a prokaryotic expression vector, pET-24a. BALB/c mice were immunized subcutaneously with purified rC-HapS. Serum IgG responses to purified rC-HapS, serum IgG subclasses were determined by ELISA and functional activity of antibodies was examined by Serum Bactericidal Assay. The output of rC-HapS was approximately 62% of the total bacterial proteins. Serum IgG responses were significantly increased in immunized group with rC-HapS mixed with Freund's adjuvant in comparison with control groups. Analysis of the serum IgG subclasses showed that the IgG1 subclass was predominant after subcutaneous immunization in BALB/c mice (IgG2a/IgG1 < 1). The sera from rC-HapS immunized animals were strongly bactericidal against nontypeable Haemophilus influenzae. These results suggest that rC-HapS may be a potential vaccine candidate for nontypeable Haemophilus influenzae. PMID:27377430

  12. The first dose of a Haemophilus influenzae type b conjugate vaccine reactivates memory B cells: evidence for extensive clonal selection, intraclonal affinity maturation, and multiple isotype switches to IgA2

    DEFF Research Database (Denmark)

    Hougs, L; Juul, L; Ditzel, H J;

    1999-01-01

    The Ab response of a healthy adult to the first dose of a Haemophilus influenzae type b capsular polysaccharide (HibCP) conjugate vaccine was studied at the level of Ig gene usage by circulating Ab-secreting cells. Forty-one IgA and 17 IgG mRNA sequences were obtained. The major part...

  13. Recombinant C-terminal 311 amino acids of HapS adhesin as a vaccine candidate for nontypeable Haemophilus influenzae: A study on immunoreactivity in Balb/C mouse.

    Science.gov (United States)

    Tabatabaee Bafroee, Akram Sadat; Siadat, Seyed Davar; Mousavi, Seyed Fazlollah; Aghasadeghi, Mohammad Reza; Khorsand, Hashem; Nejati, Mehdi; Sadat, Seyed Mehdi; Mahdavi, Mehdi

    2016-09-01

    Hap, an auto-transporter protein, is an antigenically conserved adhesion protein which is present on both typeable and nontypeable Haemophilus influenzae. This protein has central role in bacterial attachment to respiratory tract epithelial cells. A 1000bp C-terminal fragment of Hap passenger domain (HapS) from nontypeable Haemophilus influenzae was cloned into a prokaryotic expression vector, pET-24a. BALB/c mice were immunized subcutaneously with purified rC-HapS. Serum IgG responses to purified rC-HapS, serum IgG subclasses were determined by ELISA and functional activity of antibodies was examined by Serum Bactericidal Assay. The output of rC-HapS was approximately 62% of the total bacterial proteins. Serum IgG responses were significantly increased in immunized group with rC-HapS mixed with Freund's adjuvant in comparison with control groups. Analysis of the serum IgG subclasses showed that the IgG1 subclass was predominant after subcutaneous immunization in BALB/c mice (IgG2a/IgG1 < 1). The sera from rC-HapS immunized animals were strongly bactericidal against nontypeable Haemophilus influenzae. These results suggest that rC-HapS may be a potential vaccine candidate for nontypeable Haemophilus influenzae.

  14. Specific IgA and metalloproteinase activity in bronchial secretions from stable chronic obstructive pulmonary disease patients colonized by Haemophilus influenzae

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    Millares Laura

    2012-12-01

    Full Text Available Abstract Background Haemophilus influenzae is the most common colonizing bacteria of the bronchial tree in chronic obstructive pulmonary disease (COPD, and positive cultures for this potentially pathogenic microorganism (PPM has been associated with local inflammation changes that may influence the relationships between H. influenzae and the bronchial mucosa. Methods A cross-sectional analysis of stable COPD patients enrolled in the Phenotype and Course of Chronic Obstructive Pulmonary Disease (PAC-COPD Study, focusing on bronchial colonization by H. influenzae, was performed. Specific IgA against the PPM was measured by optical density, and metalloproteinase-9 (MMP-9 and tissue inhibitor of metalloproteinase-1 (TIMP-1 using ELISA in sputum samples. Levels in patients colonized by H. influenzae and non-colonized patients were compared. Results Sputum supernatant for the measurement of specific IgA against H. influenzae was available from 54 stable COPD patients, who showed levels of specific IgA significantly lower in colonized (n=21 than in non-colonized patients (n=33 (15 [4-37] versus 31 [10-75], p=0.033, Mann-Whitney U test. Proenzyme MMP-9 was measured in 44 patients, and it was higher in colonized (n=12, 1903 [1488-6699] ng/ml than in non-colonized patients (n=32, 639 [373-972] ng/ml (p Conclusions Clinically stable COPD patients colonized by H. influenzae had lower levels of specific IgA against the microorganism and higher values of the active form of MMP-9 in their sputum supernatant than non-colonized patients. Bronchial colonization by H. influenzae may cause structural changes in the extracellular matrix through a defective defense and the production of active metalloproteinases.

  15. Molecular characterisation of the clonal emergence of high-level ciprofloxacin-monoresistant Haemophilus influenzae in the Region of Southern Denmark.

    Science.gov (United States)

    Fuursted, Kurt; Hartmeyer, Gitte Nyvang; Stegger, Marc; Andersen, Paal Skytt; Justesen, Ulrik Stenz

    2016-06-01

    Haemophilus influenzae is an important human pathogen usually susceptible to quinolones. Here we report the emergence of high-level ciprofloxacin-monoresistant H. influenzae in the Region of Southern Denmark. Four isolates were collected for phenotypic and molecular characterisation using whole-genome sequencing (WGS). During an 18-month period, the occurrence of high-level ciprofloxacin-monoresistant H. influenzae in patients aged 1-77 years from sputum, ear and eye samples was detected. An epidemiological link between the patients could not be identified. The isolates were non-encapsulated, biotype III and were demonstrated by WGS to be clonal belonging to a single clade with an unknown multilocus sequence type (double-locus variant of ST196). The antibiogram demonstrated that they were all monoresistant to ciprofloxacin with a minimum inhibitory concentration (MIC) >32mg/L. In silico resistome analysis revealed identical, both previously characterised and novel, putative resistance-related mutations in gyrA (S84L and D88N), parC (K20R, S84I, D356A or T356A, and M481I) and parE (E151K, I159A, D420N and S599A) in all isolates. The isolates were otherwise negative for any resistance genes. This is the first description of the clonal emergence of high-level monoresistant H. influenzae due to amino acid substitutions in gyrA, parC and parE.

  16. Molecular characterisation of the clonal emergence of high-level ciprofloxacin-monoresistant Haemophilus influenzae in the Region of Southern Denmark.

    Science.gov (United States)

    Fuursted, Kurt; Hartmeyer, Gitte Nyvang; Stegger, Marc; Andersen, Paal Skytt; Justesen, Ulrik Stenz

    2016-06-01

    Haemophilus influenzae is an important human pathogen usually susceptible to quinolones. Here we report the emergence of high-level ciprofloxacin-monoresistant H. influenzae in the Region of Southern Denmark. Four isolates were collected for phenotypic and molecular characterisation using whole-genome sequencing (WGS). During an 18-month period, the occurrence of high-level ciprofloxacin-monoresistant H. influenzae in patients aged 1-77 years from sputum, ear and eye samples was detected. An epidemiological link between the patients could not be identified. The isolates were non-encapsulated, biotype III and were demonstrated by WGS to be clonal belonging to a single clade with an unknown multilocus sequence type (double-locus variant of ST196). The antibiogram demonstrated that they were all monoresistant to ciprofloxacin with a minimum inhibitory concentration (MIC) >32mg/L. In silico resistome analysis revealed identical, both previously characterised and novel, putative resistance-related mutations in gyrA (S84L and D88N), parC (K20R, S84I, D356A or T356A, and M481I) and parE (E151K, I159A, D420N and S599A) in all isolates. The isolates were otherwise negative for any resistance genes. This is the first description of the clonal emergence of high-level monoresistant H. influenzae due to amino acid substitutions in gyrA, parC and parE. PMID:27436470

  17. Multiplex quantitative PCR for detection of lower respiratory tract infection and meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae and Neisseria meningitidis

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    Welinder-Olsson Christina

    2010-12-01

    Full Text Available Abstract Background Streptococcus pneumoniae and Haemophilus influenzae cause pneumonia and as Neisseria meningitidis they are important agents of meningitis. Although several PCR methods have been described for these bacteria the specificity is an underestimated problem. Here we present a quantitative multiplex real-time PCR (qmPCR for detection of S. pneumoniae (9802 gene fragment, H. influenzae (omp P6 gene and N. meningitidis (ctrA gene. The method was evaluated on bronchoalveolar lavage (BAL samples from 156 adults with lower respiratory tract infection (LRTI and 31 controls, and on 87 cerebrospinal fluid (CSF samples from meningitis patients. Results The analytical sensitivity was not affected by using a combined mixture of reagents and a combined DNA standard (S. pneumoniae/H. influenzae/N. meningitidis in single tubes. By blood- and BAL-culture and S. pneumoniae urinary antigen test, S. pneumoniae and H. influenzae were aetiological agents in 21 and 31 of the LTRI patients, respectively. These pathogens were identified by qmPCR in 52 and 72 of the cases, respectively, yielding sensitivities and specificities of 95% and 75% for S. pneumoniae, and 90% and 65% for H. influenzae, respectively. When using a cut-off of 105 genome copies/mL for clinical positivity the sensitivities and specificities were 90% and 80% for S. pneumoniae, and 81% and 85% for H. influenzae, respectively. Of 44 culture negative but qmPCR positive for H. influenzae, 41 were confirmed by fucK PCR as H. influenzae. Of the 103 patients who had taken antibiotics prior to sampling, S. pneumoniae and H. influenzae were identified by culture in 6% and 20% of the cases, respectively, and by the qmPCR in 36% and 53% of the cases, respectively. In 87 CSF samples S. pneumoniae and N. meningitidis were identified by culture and/or 16 S rRNA in 14 and 10 samples and by qmPCR in 14 and 10 samples, respectively, giving a sensitivity of 100% and a specificity of 100% for both

  18. Characterization of the N-Acetyl-5-neuraminic Acid-binding Site of the Extracytoplasmic Solute Receptor (SiaP) of Nontypeable Haemophilus influenzae Strain 2019

    Energy Technology Data Exchange (ETDEWEB)

    Johnston, Jason W.; Coussens, Nathan P.; Allen, Simon; Houtman, Jon C.D.; Turner, Keith H.; Zaleski, Anthony; Ramaswamy, S.; Gibson, Bradford W.; Apicella, Michael A. (Iowa); (Buck Inst.)

    2012-11-14

    Nontypeable Haemophilus influenzae is an opportunistic human pathogen causing otitis media in children and chronic bronchitis and pneumonia in patients with chronic obstructive pulmonary disease. The outer membrane of nontypeable H. influenzae is dominated by lipooligosaccharides (LOS), many of which incorporate sialic acid as a terminal nonreducing sugar. Sialic acid has been demonstrated to be an important factor in the survival of the bacteria within the host environment. H. influenzae is incapable of synthesizing sialic acid and is dependent on scavenging free sialic acid from the host environment. To achieve this, H. influenzae utilizes a tripartite ATP-independent periplasmic transporter. In this study, we characterize the binding site of the extracytoplasmic solute receptor (SiaP) from nontypeable H. influenzae strain 2019. A crystal structure of N-acetyl-5-neuraminic acid (Neu5Ac)-bound SiaP was determined to 1.4 {angstrom} resolution. Thermodynamic characterization of Neu5Ac binding shows this interaction is enthalpically driven with a substantial unfavorable contribution from entropy. This is expected because the binding of SiaP to Neu5Ac is mediated by numerous hydrogen bonds and has several buried water molecules. Point mutations targeting specific amino acids were introduced in the putative binding site. Complementation with the mutated siaP constructs resulted either in full, partial, or no complementation, depending on the role of specific residues. Mass spectrometry analysis of the O-deacylated LOS of the R127K point mutation confirmed the observation of reduced incorporation of Neu5Ac into the LOS. The decreased ability of H. influenzae to import sialic acid had negative effects on resistance to complement-mediated killing and viability of biofilms in vitro, confirming the importance of sialic acid transport to the bacterium.

  19. Quantitative detection of Staphylococcus aureus, Streptococcus pneumoniae and Haemophilus influenzae in patients with new influenza A (H1N1)/2009 and influenza A/2010 virus infection.

    Science.gov (United States)

    Safaeyan, Firouzeh; Nahaei, Mohammad Reza; Seifi, Sirus Jedary; Kafil, Hossein Samadi; Sadeghi, Javid

    2015-01-01

    Einleitung: Die Virusgrippe ist eine saisonale Infektionskrankheit, die mit ausgeprägterer Morbidität und Mortalität einhergeht. In den USA werden jährlich mehr als 35.000 Todesfälle und 200.000 Krankenhausbehandlungen erfasst. Die mit der viralen Primärinfektion assoziierte bakterielle Superinfektion oder Ko-Infektion verursacht schwere Krankheitsverläufe speziell bei Hochrisikogruppen wie alten Menschen und Kleinkindern. Zielsetzung: Die Zielsetzung der Studie bestand in der quantitativen Bestimmung von S. aureus, S. pneumoniae and H. influenzae bei Patienten mit saisonaler Influenza A bzw. pandemischer Influenza A (H1N1) und Symptomen respiratorischer Infektionen, im Vergleich zu je einer Kontrollgruppe. Methode: Insgesamt wurden von April 2009 bis April 2010 625 Patienten mit Verdacht auf eine respiratorische Infektion untersucht, davon 58 Patienten mit Nachweis von Influenza A (H1N1). Vom November 2010 bis zum Februar 2011 wurden 158 Patienten mit respiratorischen Symptomen auf das Vorkommen der saisonalen Influenza A untersucht, davon erwiesen sich 25 als positiv. Zur Ermittlung der bakteriellen Kolonisation wurden parallel 62 gesunde Personen untersucht (Kontrollgruppe). Bei der verwendeten Real-time PCR wurde als Cutoff zur Unterscheidung von Koloniation und Infektion im Respirationstrakt 10(4) CFU/ml eingeführt. Ergebnisse:S. aureus, S. pneumoniae und H. influenzae wurden bei 12%, 26% bzw. 33% der Patienten mit Nachweis von Influenzavirus A (H1N1) gefunden; die Häufigkeit in der Kontrollgruppe betrug 9%, 19% bzw. 31%. Bei der saisonalen Influenza A waren bei 12%, 24% bzw. 32% die Erreger nachweisbar, in der parallelen Kontrollgruppe bei 5%, 11% bzw. 10%. Schlussfolgerung: Die Ergebnisse zeigen, dass der Serotyp der pandemischen Influenza A (H1N1) die Inzidenz der bakteriellen Superinfektion für die drei untersuchten Bakterienspecies nicht erhöht hat. Die quantitative Detektion einer sekundären bakteriellen Infektion mittels Real-time PCR ist

  20. [Multilocus sequence-typing for characterization of Moscow strains of Haemophilus influenzae type b].

    Science.gov (United States)

    Platonov, A E; Mironov, K O; Iatsyshina, S B; Koroleva, I S; Platonova, O V; Gushchin, A E; Shipulin, G A

    2003-01-01

    Haemophilius influenzae, type b (Hib) bacteria, were genotyped by multilocus sequence typing (MLST) using 5 loci (adk, fucK, mdh, pgi, recA). 42 Moscow Hib strains (including 38 isolates form cerebrospinal fluid of children, who had purulent meningitis in 1999-2001, and 4 strains isolated from healthy carriers of Hib), as well as 2 strains from Yekaterinburg were studied. In MLST a strain is characterized, by alleles and their combinations (an allele profile) referred to also as sequence-type (ST). 9 Sts were identified within the Russian Hib bacteria: ST-1 was found in 25 strains (57%), ST-12 was found in 8 strains (18%), ST-11 was found in 4 strains (9%) and ST-15 was found in 2 strains (4.5%); all other STs strains (13, 14, 16, 17, 51) were found in isolated cases (2.3%). A comparison of allelic profiles and of nucleotide sequences showed that 93% of Russian isolates, i.e. strain with ST-1, 11, 12, 13, 15 and 17, belong to one and the same clonal complex. 2 isolates from Norway and Sweden from among 7 foreign Hib strains studied up to now can be described as belonging to the same clonal complex; 5 Hib strains were different from the Russian ones. PMID:12800772

  1. 成人呼吸道感染流感嗜血杆菌临床特点与耐药性研究%Characteristics and Drug Resistance of Respiratory Tract Infections for Haemophilus influenzae in Adult Patients

    Institute of Scientific and Technical Information of China (English)

    孙潺

    2015-01-01

    Objective To investigate the characteristics and drug resistance of respiratory tract infection in adult respiratory tract infection for Haemophilus influenzae.MethodsSelected 66 cases from March 2013 to July 2014 in our hospital with adult respiratory tract infection as the research object,and implemented Haemophilus influenzae detection and Haemophilus influenzae culture. ResultsThere was a close relationship between the sex and age of the patients with influenza. Haemophilus influenzae to ampicilin,cotrimoxazole,tetracycline showed high resistance.Conclusion It is important to strengthen the research of drug resistance of influenza.%目的:探讨成人呼吸道感染流感嗜血杆菌特点及耐药性。方法选择2013年3月~2014年7月在我院接受治疗的66例流感嗜血杆菌成人呼吸道感染患者作为研究对象,对患者痰液标本实施流感嗜血杆菌检测和嗜血杆菌分离培养。结果流感嗜血杆菌感染与患者性别、年龄等均存在密切联系。流感嗜血杆菌对氨苄西林、复方新诺明、四环素均表现出较高耐药性。结论临床需要加强对流感嗜血杆菌的耐药性研究。

  2. The Epidemiology of Invasive Haemophilus influenzae Non-Serotype B Disease in Ontario, Canada from 2004 to 2013.

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    Shalini Desai

    Full Text Available Since the widespread use of Haemophilus influenzae (Hi type b (Hib vaccines among children aged <5 years, an increase in invasive non-Hib disease incidence has been reported internationally. We sought to describe the epidemiology of invasive non-Hib disease in Ontario, Canada (population ~13.5 million.Confirmed invasive non-Hib cases (non-typeable [NTHi] and serotypes a, c, d, e, and f were obtained from the provincial laboratory data system from 2004-2013. Data were deterministically linked to the provincial reportable disease system to provide further case information. Antibiotic resistance data were analysed separately from 2010-2014. Descriptive analyses included incidence rates, age group, serotype, site of specimen collection and resistance patterns; ethnicity data were not available. Temporal trends were evaluated by Poisson regression and p-values <0.05 were considered significant.A total of 1307 cases of invasive non-Hib disease were included, increasing from 0.67 cases to 1.60 cases /100,000 from 2004 to 2013. Significant increases in the incidence of NTHi (0.50 to 1.28 cases/100 000 population, Hia (0.02 to 0.08 cases/100, 000 and Hif (0.13 to 0.18 cases/100, 000 population were seen. Among persons aged 40-64 years, 3 Hi strains significantly increased over time; NTHi (0.22 to 0.99 cases/100, 000, Hia (0.00 to 0.06 cases/100, 000 and Hif (0.05 to 0.21 cases/100, 000. Among persons aged 65-84 years, there was a significant increase of NTHi (1.62 to 3.14 cases/100, 000 and Hia (0.00 to 0.34 cases/100, 000. Among persons aged 85+ years, only NTHi significantly increased from 4.89 to 10.28 cases/100, 000. Antimicrobial resistance (AMR to ampicillin and clarithromycin was seen in greater than 25% of isolates but AMR did not increase over the duration of this study.The incidence of invasive non-Hib disease has increased over time; NTHi, Hif and Hia are emerging pathogens, and should be monitored.

  3. Study on genotyping of Haemophilus influenzae%流感嗜血杆菌基因分型研究

    Institute of Scientific and Technical Information of China (English)

    温群文; 刘楚云; 鞠长燕; 俞慕华; 田国忠; 段永翔

    2015-01-01

    目的 应用生物学分型、血清学分型和脉冲场凝胶电泳(Pulsed-field gel electrophoresis,PFGE)分析从健康人群分离的流感嗜血杆菌(Haemophilus influenzae,Hi)的分子流行病学规律以及与Hi生物学分型、血清学分型、青酶素酶之间的关系.方法 对深圳市1月龄以上380名健康人群采集咽拭子,共分离培养175株流感嗜血杆菌,参照Pitt-man方法对菌株进行血清学分型,应用PCR方法进行流感嗜血杆菌属及荚膜鉴定,参照Kilian方法对其中的141株菌株进行生物学分型,从不同年龄组选出共59株流感嗜血杆菌株,应用PFGE技术进行分子分型研究.结果 141株流感嗜血杆菌分为8个生物型,Ⅰ~Ⅷ型分别为8.51%、36.88%、34.04%、4.96%、7.80%、0.71%、6.38%、0.71%.分离的菌株以无荚膜的不可分型Hi为主(97.14%),5株为有荚膜型,其中2株为b型,3株为e型,Hi产青霉素酶的阳性率为14.18%,59株Hi菌株分为52个PFGE基因型.结论 深圳市健康人群Hi携带菌株多为无荚膜的不可分型流感嗜血杆菌,生物学分型以Ⅱ、Ⅲ型为主.PGFE基因分型呈明显的多态性.

  4. 2010-2012年流感嗜血杆菌耐药性监测%2010-2012 surveillance of antimicrobial resistance in haemophilus influenzae

    Institute of Scientific and Technical Information of China (English)

    龙姗姗; 喻华

    2014-01-01

    Objective To investigate the distribution and antimicrobial resistance of haemophilus influenzae ,and to pro-vide reference for reasonable use of antibiotics .Methods Clinical isolates from patients in our hospital from January 2010 to December 2012 were identified by VITEK2-compact automatic identification system , bacterial susceptibility tests were per-formed on strains by ATB system , the results were determined according to the guidelines of CLSI in 2012,and β-lactamase was detected by nitrocefin disk diffusion method .WHONET5.6 software was applied for data analysis .Results A total of 201 strains of haemophilus influenzae were obtained from our hospital in recent three years .Among them,193 strains were from sputum specimen (96.0%),6 from bronchoalveolar lavage fluid specimen (2.9%),2 from blood specimen (1.1%).As for the distribution of haemophilus influenzae in clinical departments ,the top three were respiratory department (104 strains, 51. 7%),surgical department (31 strains,15.4%) and ICU (27 strains,13.4%).The results of bacterial susceptibility tests showed that haemophilus influenzae was sensitive to rifampicin , ampicillin,chloramphenicol and cotrimoxazole with a rate of 45.5%,73.4%,87.5%and 87.1%respectively,and that the sensitive rate of haemophilus influenzae to amoxicillin /clavu-lanic acid,cefotaxime,cefaclor,tetracycline, cefuroxime and ofloxacin was higher than 90%.Isolates from adults were more sensitive to ampicillin and rifampicin than those from children ,but for tetracycline , isolates from children were more sensitive than those from adults .52 strains were positive to β-lactamase with a rate of 25.9%, and the rate of β-lactamase positive strains from children was significantly higher than that from adults .Conclusion Haemophilus influenzae has a higher resistant rate to rifampicin.Amoxicillin /clavulanate,cefotaxime,cefaclor,cefuroxime and ofloxacin are still the effective antibiotics for the control of haemophilus influenzae

  5. Proteomic expression profiling of Haemophilus influenzae grown in pooled human sputum from adults with chronic obstructive pulmonary disease reveal antioxidant and stress responses

    Directory of Open Access Journals (Sweden)

    Brauer Aimee L

    2010-06-01

    Full Text Available Abstract Background Nontypeable Haemophilus influenzae colonizes and infects the airways of adults with chronic obstructive pulmonary disease, the fourth most common cause of death worldwide.Thus, H. influenzae, an exclusively human pathogen, has adapted to survive in the hostile environment of the human airways.To characterize proteins expressed by H. influenzae in the airways, a prototype strain was grown in pooled human sputum to simulate conditions in the human respiratory tract.The proteins from whole bacterial cell lysates were solubilized with a strong buffer and then quantitatively cleaned with an optimized precipitation/on-pellet enzymatic digestion procedure.Proteomic profiling was accomplished by Nano-flow liquid chromatography/mass spectroscopy with low void volume and high separation efficiency with a shallow, long gradient. Results A total of 1402 proteins were identified with high confidence, including 170 proteins that were encoded by genes that are annotated as conserved hypothetical proteins.Thirty-one proteins were present in greater abundance in sputum-grown conditions at a ratio of > 1.5 compared to chemically defined media.These included 8 anti-oxidant and 5 stress-related proteins, suggesting that expression of antioxidant activity and stress responses is important for survival in the airways.Four proteins involved in uptake of divalent anions and 9 proteins that function in uptake of various molecules were present in greater abundance in sputum-grown conditions. Conclusions Proteomic expression profiling of H. influenzae grown in pooled human sputum revealed increased expression of antioxidant, stress-response proteins and cofactor and nutrient uptake systems compared to media grown cells.These observations suggest that H. influenzae adapts to the oxidative and nutritionally limited conditions of the airways in adults with chronic obstructive pulmonary disease by increasing expression of molecules necessary for survival

  6. Identification of Haemophilus influenzae Type b Isolates by Use of Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry

    Science.gov (United States)

    Månsson, Viktor; Kostrzewa, Markus; Nilson, Bo; Riesbeck, Kristian

    2015-01-01

    Haemophilus influenzae type b (Hib) is, in contrast to non-type b H. influenzae, associated with severe invasive disease, such as meningitis and epiglottitis, in small children. To date, accurate H. influenzae capsule typing requires PCR, a time-consuming and cumbersome method. Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) provides rapid bacterial diagnostics and is increasingly used in clinical microbiology laboratories. Here, MALDI-TOF MS was evaluated as a novel approach to separate Hib from other H. influenzae. PCR-verified Hib and non-Hib reference isolates were selected based on genetic and spectral characteristics. Mass spectra of reference isolates were acquired and used to generate different classification algorithms for Hib/non-Hib differentiation using both ClinProTools and the MALDI Biotyper software. A test series of mass spectra from 33 Hib and 77 non-Hib isolates, all characterized by PCR, was used to evaluate the algorithms. Several algorithms yielded good results, but the two best were a ClinProTools model based on 22 separating peaks and subtyping main spectra (MSPs) using MALDI Biotyper. The ClinProTools model had a sensitivity of 100% and a specificity of 99%, and the results were 98% reproducible using a different MALDI-TOF MS instrument. The Biotyper subtyping MSPs had a sensitivity of 97%, a specificity of 100%, and 93% reproducibility. Our results suggest that it is possible to use MALDI-TOF MS to differentiate Hib from other H. influenzae. This is a promising method for rapidly identifying Hib in unvaccinated populations and for the screening and surveillance of Hib carriage in vaccinated populations. PMID:25926500

  7. Origin of the diversity in DNA recognition domains in phasevarion associated modA genes of pathogenic Neisseria and Haemophilus influenzae.

    Science.gov (United States)

    Gawthorne, Jayde A; Beatson, Scott A; Srikhanta, Yogitha N; Fox, Kate L; Jennings, Michael P

    2012-01-01

    Phase variable restriction-modification (R-M) systems have been identified in a range of pathogenic bacteria. In some it has been demonstrated that the random switching of the mod (DNA methyltransferase) gene mediates the coordinated expression of multiple genes and constitutes a phasevarion (phase variable regulon). ModA of Neisseria and Haemophilus influenzae contain a highly variable, DNA recognition domain (DRD) that defines the target sequence that is modified by methylation and is used to define modA alleles. 18 distinct modA alleles have been identified in H. influenzae and the pathogenic Neisseria. To determine the origin of DRD variability, the 18 modA DRDs were used to search the available databases for similar sequences. Significant matches were identified between several modA alleles and mod gene from distinct bacterial species, indicating one source of the DRD variability was via horizontal gene transfer. Comparison of DRD sequences revealed significant mosaicism, indicating exchange between the Neisseria and H. influenzae modA alleles. Regions of high inter- and intra-allele similarity indicate that some modA alleles had undergone recombination more frequently than others, generating further diversity. Furthermore, the DRD from some modA alleles, such as modA12, have been transferred en bloc to replace the DRD from different modA alleles. PMID:22457715

  8. Origin of the diversity in DNA recognition domains in phasevarion associated modA genes of pathogenic Neisseria and Haemophilus influenzae.

    Science.gov (United States)

    Gawthorne, Jayde A; Beatson, Scott A; Srikhanta, Yogitha N; Fox, Kate L; Jennings, Michael P

    2012-01-01

    Phase variable restriction-modification (R-M) systems have been identified in a range of pathogenic bacteria. In some it has been demonstrated that the random switching of the mod (DNA methyltransferase) gene mediates the coordinated expression of multiple genes and constitutes a phasevarion (phase variable regulon). ModA of Neisseria and Haemophilus influenzae contain a highly variable, DNA recognition domain (DRD) that defines the target sequence that is modified by methylation and is used to define modA alleles. 18 distinct modA alleles have been identified in H. influenzae and the pathogenic Neisseria. To determine the origin of DRD variability, the 18 modA DRDs were used to search the available databases for similar sequences. Significant matches were identified between several modA alleles and mod gene from distinct bacterial species, indicating one source of the DRD variability was via horizontal gene transfer. Comparison of DRD sequences revealed significant mosaicism, indicating exchange between the Neisseria and H. influenzae modA alleles. Regions of high inter- and intra-allele similarity indicate that some modA alleles had undergone recombination more frequently than others, generating further diversity. Furthermore, the DRD from some modA alleles, such as modA12, have been transferred en bloc to replace the DRD from different modA alleles.

  9. Nonencapsulated or nontypeable Haemophilus influenzae are more likely than their encapsulated or serotypeable counterparts to have mutations in their fucose operon.

    Science.gov (United States)

    Shuel, Michelle L; Karlowsky, Kathleen E; Law, Dennis K S; Tsang, Raymond S W

    2011-12-01

    Population biology of Haemophilus influenzae can be studied by multilocus sequence typing (MLST), and isolates are assigned sequence types (STs) based on nucleotide sequence variations in seven housekeeping genes, including fucK. However, the ST cannot be assigned if one of the housekeeping genes is absent or cannot be detected by the current protocol. Occasionally, strains of H. influenzae have been reported to lack the fucK gene. In this study, we examined the prevalence of this mutation among our collection of H. influenzae isolates. Of the 704 isolates studied, including 282 encapsulated and 422 nonencapsulated isolates, nine were not typeable by MLST owing to failure to detect the fucK gene. All nine fucK-negative isolates were nonencapsulated and belonged to various biotypes. DNA sequencing of the fucose operon region confirmed complete deletion of genes in the operon in seven of the nine isolates, while in the remaining two isolates, some of the genes were found intact or in parts. The significance of these findings is discussed. PMID:22107351

  10. Origin of the diversity in DNA recognition domains in phasevarion associated modA genes of pathogenic Neisseria and Haemophilus influenzae.

    Directory of Open Access Journals (Sweden)

    Jayde A Gawthorne

    Full Text Available Phase variable restriction-modification (R-M systems have been identified in a range of pathogenic bacteria. In some it has been demonstrated that the random switching of the mod (DNA methyltransferase gene mediates the coordinated expression of multiple genes and constitutes a phasevarion (phase variable regulon. ModA of Neisseria and Haemophilus influenzae contain a highly variable, DNA recognition domain (DRD that defines the target sequence that is modified by methylation and is used to define modA alleles. 18 distinct modA alleles have been identified in H. influenzae and the pathogenic Neisseria. To determine the origin of DRD variability, the 18 modA DRDs were used to search the available databases for similar sequences. Significant matches were identified between several modA alleles and mod gene from distinct bacterial species, indicating one source of the DRD variability was via horizontal gene transfer. Comparison of DRD sequences revealed significant mosaicism, indicating exchange between the Neisseria and H. influenzae modA alleles. Regions of high inter- and intra-allele similarity indicate that some modA alleles had undergone recombination more frequently than others, generating further diversity. Furthermore, the DRD from some modA alleles, such as modA12, have been transferred en bloc to replace the DRD from different modA alleles.

  11. 北京和天津两地在校学生流感嗜血杆菌携带情况监测%The carriage of haemophilus influenzae in students from Beijing and Tianjin

    Institute of Scientific and Technical Information of China (English)

    张铁钢; 罗明; 龚成; 吕敏; 丁小静; 牛艳芬; 吴疆

    2012-01-01

    Objective:To monitor the carriage of Haemophilus influenza in students from Beijing and Tianjin in 2009 and 2010. Methods: In Beijing and Tianjin, the clinical specimens were collected from the school students with acute upper respiratory tract infection. The nucleic acids detection and serotyping of haemophilus influenzae and serotyping were performed with molecular technology. Results: 154 cases including 84 from Beijing and 70 from Tianjin were surveied. 80 cases were positive with haemophilus influenzae nucleic acid in Beijing and 61 cases were positive with haemophilus influenzae nucleic acid in Tianjin. In 141 positive clinical specimens, capsulate and non - capsulate haemophilus influenzae were found in 35 cases and 106 cases, respectively. In 35 cases with capsulate type, 28 specimens were c type and 1 was d type, the rest could not be typed. Conclusion: The noncapsulate type of Haemophilus influenza were mainly carried by the students. In encapsulated strains, c type was the major type carried by the students in Beijing and Tianjin.%目的:监测2009年/2010年北京市和天津市2地在校学生流感嗜血杆菌携带情况.方法:选择北京市和天津市大中小学生上呼吸道感染患者作为监测对象,利用PCR分子生物学技术,对急性上呼吸道患者临床标本进行流感嗜血杆菌核酸检测和血清型分型.结果:共调查了154例上感病例,其中北京84例,天津70例.北京病例标本中共检出流感嗜血杆菌80例,天津病例标本中共检出流感嗜血杆菌61例.141例阳性标本中,非荚膜型106例,荚膜型35例.35例荚膜型中,c型28例,d型1例,另外6例未能分型.结论:北京和天津2地学生主要携带非荚膜型流感嗜血杆菌,携带的荚膜型菌株主要为c型流感嗜血杆菌.

  12. Artritis séptica de cadera por Haemophilus influenzae tipo F en la infancia: a propósito de un caso

    Directory of Open Access Journals (Sweden)

    N. Ulibarrena

    2013-04-01

    Full Text Available La artritis séptica de cadera es una infección aguda, predominantemente de etiología bacteriana. A pesar de que el patógeno principal en cualquier grupo de edad es Staphylococcus aureus, existen otros microorganismos potencialmente patógenos. Describimos el caso de una paciente de 18 meses con artritis séptica de cadera por un agente causal infrecuente (Haemophilus influenzae tipo f de buena evolución. Este caso pone de manifiesto la importancia de considerar otros agentes etiológicos menos frecuentes en las artritis sépticas infantiles, realizando artrocentesis con cultivo de líquido articular antes de iniciar el tratamiento antibiótico, siempre que sea posible. Además, como ocurrió en este caso, conviene tener en cuenta que el resultado positivo del cultivo del líquido sinovial para Haemophilus influezae obliga a tipificar la bacteria para descartar o confirmar un fracaso vacunal.

  13. Minimal biofilm eradication concentration of antimicrobial agents against nontypeable Haemophilus influenzae isolated from middle ear fluids of intractable acute otitis media.

    Science.gov (United States)

    Takei, Shin; Hotomi, Muneki; Yamanaka, Noboru

    2013-06-01

    Nontypeable Haemophilus influenzae (NTHi) makes the clinical course of acute otitis media (AOM) intractable by forming a biofilm that may hamper the clearance of the bacteria from middle ear cavity. In this study, we evaluated the minimum biofilm eradication concentration (MBEC) of antimicrobial agents against biofilm-forming NTHi strains. Twelve NTHi strains isolated from middle ear fluids of Japanese children with intractable AOM before antimicrobial treatment were evaluated for MBEC of fluoroquinolones in comparison with those of β-lactams and macrolides. AMPC and CDTR required much higher concentration, i.e., high MBECs, to suppress the biofilm formation of NTHi. In contrast, fluoroquinolones followed by macrolides showed lower MBECs. MBEC would be a good parameter to infer the efficacies of antimicrobials against NTHi in biofilm.

  14. Children with Haemophilus influenzae type b (Hib) vaccine failure have long-term bactericidal antibodies against virulent Hib strains with multiple capsular loci.

    Science.gov (United States)

    Townsend-Payne, Kelly; Ladhani, Shamez N; Findlow, Helen; Slack, Mary; Borrow, Ray

    2016-07-25

    Children who develop invasive Haemophilus influenzae serotype b (Hib) disease after immunisation with a highly-effective conjugate vaccine are more likely to have been infected with Hib strains possessing multiple copies of the capsulation locus. Using a recently-validated serum bactericidal antibody (SBA) assay, we tested convalescent sera from 127 Hib vaccine failure cases against clinical Hib strains expressing 1-5 copies of the capsulation locus. SBA titres correlated weakly with anti-capsular IgG antibody concentrations and there was no association between SBA geometric mean titres and number of capsulation locus copies. After infection, children with Hib vaccine failure were equally protected against Hib strains with 1-5 copies of the capsulation locus. PMID:27317452

  15. The estimation of safety booster vaccination children is more senior than 1,5 years against diphtheria, perussis, tetanus, poliomyelitis and Haemophilus influenzae type B with Pentaxim

    Directory of Open Access Journals (Sweden)

    S. M. Harit

    2009-01-01

    Full Text Available Supervision over 200 children in the age of 18–42 months (64 healthy and 136 with allergic displays, defeats CNS, often ill, and also having in the anamnesis of reaction to previous introduction DTP or Infanriks, revaccination with Pentaxim. It is shown, that 77% from them had asymptomatic current, only in 1,5% of cases strong reactions with temperature above 38,6°С were observed. Local reactions were marked at 25,5%, essentially more often at children with an allergy, defeat CNS and often ill, than at healthy (7,8%, but did not one child was not adverse events. The estimation safety vaccines Pentaxim confirms expediency of application as 1 revaccination against perussis, diphtheria, tetanus, poliomyelitis and unitary vaccination against Haemophilus influenzae type B in children are more senior than year with a various state of health.

  16. Non-epitope-specific suppression of the antibody response to Haemophilus influenzae type b conjugate vaccines by preimmunization with vaccine components

    DEFF Research Database (Denmark)

    Barington, T; Skettrup, M; Juul, L;

    1993-01-01

    Recently, conjugate vaccines containing Haemophilus influenzae type b capsular polysaccharide (HibCP) coupled to protein carriers were introduced for use in infants and certain adult risk groups. Similar conjugate vaccines against other capsulated bacteria are currently under development for both...... children and adults. Despite its potential importance, the possible influence of preexisting immunity to the components of such conjugates on the vaccination response in humans has been addressed by few studies. To study this issue, we randomized 82 healthy adult volunteers into six groups and vaccinated...... them twice, with a 4-week interval between immunizations. Four groups received tetanus toxoid (TT) or diphtheria toxoid (DT) and then HibCP coupled to TT (HibCP-TT) or DT (HibCP-DT). Two groups received HibCP-TT followed by HibCP-DT or vice versa. The total antibody levels to HibCP, TT, and DT...

  17. Opposite effects of actively and passively acquired immunity to the carrier on responses of human infants to a Haemophilus influenzae type b conjugate vaccine

    DEFF Research Database (Denmark)

    Barington, T; Gyhrs, A; Kristensen, Kim;

    1994-01-01

    hundred forty-four infants were vaccinated with HibCP-TT at 5 and 6 months. They were randomized into three groups that received TT as part of a diphtheria-tetanus-polio vaccine at either 6 and 7 months (group A), 5 and 6 months (group B), or 4 and 5 months (group C). Maternally acquired TT antibodies......Vaccination of infants with Haemophilus influenzae type b (Hib) capsular polysaccharide (HibCP) coupled to carrier proteins has proven protective against invasive Hib diseases in several trials. However, insufficient immunogenicity has been noted in certain populations. Therefore, studies analyzing...... factors influencing the antibody response to conjugate vaccines are needed. In this study, the response to HibCP coupled to tetanus toxoid (TT) was examined in relation to (i) priming with or coadministration of the carrier protein and (ii) the levels of passively acquired maternal TT antibodies. One...

  18. Relation between enzyme-linked immunosorbent assay and radioimmunoassay for detection of antibodies to the capsular polysaccharide of Haemophilus influenzae type b

    Energy Technology Data Exchange (ETDEWEB)

    Kristensen, K. (Streptococcus Department, Statens Seruminstitut, Copenhagen (Denmark)); Weis Bentzon, M. (Department of Biostatistics, Statens Seruminstitut, Copenhagen (Denmark))

    1992-01-01

    The measurement of antibodies to the capsular polysaccharide (PRP) of Haemophilus influenzae type b (Hib) is important because vaccines inducing such antibodies are now available. We developed and evaluated an enzyme-linked immunosorbent assay (ELISA) for detection of these antibodies based on direct coating of the plates with tyraminated PRP. The assay fulfilled the requirements for parallel line assays; it was sensitive, specific, and reproducible with a coefficient of variation between days of 19%. Results from the ELISA were compared with results from radioimmunoassay and a correlation coefficient of 0.93 was found. Results obtained by the two methods were proportional and the relation was indepenedent of the antibody level. The relation between them was also unaffected by the contribution of different antibody isotypes, indicating that these were measured to the same extent by both methods. ELISA employing direct coating of the plates with tyraminated PRP represents a useful alternative for detection of antibodies when studying immunogenicity of Hib vaccines. (au).

  19. Children with Haemophilus influenzae type b (Hib) vaccine failure have long-term bactericidal antibodies against virulent Hib strains with multiple capsular loci.

    Science.gov (United States)

    Townsend-Payne, Kelly; Ladhani, Shamez N; Findlow, Helen; Slack, Mary; Borrow, Ray

    2016-07-25

    Children who develop invasive Haemophilus influenzae serotype b (Hib) disease after immunisation with a highly-effective conjugate vaccine are more likely to have been infected with Hib strains possessing multiple copies of the capsulation locus. Using a recently-validated serum bactericidal antibody (SBA) assay, we tested convalescent sera from 127 Hib vaccine failure cases against clinical Hib strains expressing 1-5 copies of the capsulation locus. SBA titres correlated weakly with anti-capsular IgG antibody concentrations and there was no association between SBA geometric mean titres and number of capsulation locus copies. After infection, children with Hib vaccine failure were equally protected against Hib strains with 1-5 copies of the capsulation locus.

  20. Immunogenicity of a combined DTPa-HB vaccine co-administered with Haemophilus influenzae type B conjugate vaccine (PRP-T for primary and booster vaccinations

    Directory of Open Access Journals (Sweden)

    Humberto Bracco Neto

    2005-10-01

    Full Text Available OBJECTIVE: To evaluate the immunogenicity of a combined DTPa-HB vaccine co-administered with Haemophilus influenzae type b conjugate vaccine (PRP-T in Brazilian infants. MATERIAL AND METHODS: A prospective and open clinical study, in which 110 infants were immunized with a three-dose primary vaccination regime at two, four and six months of age and with a single booster vaccination. Blood samples were drawn immediately before the first dose, one month after the third dose, at the time of the booster dose and one month after the booster to assess seropositivity and antibody geometric mean titers (GMTs of antibodies for diphtheria, tetanus, hepatitis B, Haemophilus influenzae type b and for the three pertussis antigens: Pertussis Toxin (PT, Filamentous Hemagglutinin (FHA and Pertactin (PRN. RESULTS: Among the original 110 infants, 93 completed the study. Seropositivity was 100% for all seven involved antibodies, after the primary vaccination course. At the time of the booster dose, all antibodies (except diphtheria 33.7% and anti-PT 59% were seropositive for more than 94% of subjects. After the booster, seropositivity increased to 100% for all antibodies. The GMT of these antibodies followed a similar pattern, with a strong increase after the primary course, followed by a second increase after the booster dose. At this time, GMT was2- to 7-fold higher than after the primary course, for all vaccine components. CONCLUSIONS: Concomitant administration of DTPa-HB and Hib vaccines elicited strong seroprotection for all the antigenic components. No interference with antibody response was evident. The vaccines provided high immunogenicity, following both the primary vaccinations and the booster dose.

  1. Streptococcus pneumoniae and Haemophilus influenzae as etiological agents of conjunctivitis outbreaks in the region of Ribeirão Preto, SP, Brazil

    Directory of Open Access Journals (Sweden)

    Marta I. C. MEDEIROS

    1998-01-01

    Full Text Available In the study of conjunctivitis outbreaks occurring from September 1994 to September 1996 in the region of Ribeirão Preto, conjunctival exudates of 92 patients were cultivated in Instituto Adolfo Lutz Laboratory I, Ribeirão Preto. Most cases occurred in the age range 2-7 years. The etiological agents which were most frequently isolated from the analyzed cases were: Streptococcus pneumoniae and Haemophilus influenzae, in 40.22% and 21.74%, respectively. 51.35% of the S. pneumoniae isolated strains were not typable. The oxacillin-resistant S. pneumoniae strains were submitted to the minimum inhibitory concentration test (MIC and three of them presented intermediate resistance, whereas only one was highly resistant to penicillin.No estudo de surtos de conjuntivite ocorridos no período de setembro de 1994 a setembro de 1996, na região de Ribeirão Preto, foram semeadas no Instituto Adolfo Lutz Laboratório I, Ribeirão Preto, exsudatos conjuntivais de 92 pacientes, sendo que a maioria dos casos estava na faixa etária de 2-7 anos. Os agentes etiológicos mais freqüentemente isolados dos casos analisados foram: Streptococcus pneumoniae e Haemophilus influenzae em 40,22% e 21,74% respectivamente. 51,35% das cepas de S. pneumoniae isoladas foram não tipáveis. As cepas de S. pneumoniae oxacilina resistente foram submetidas ao teste de concentração inibitória mínima (CIM, sendo que três apresentaram resistência intermediária e apenas uma foi altamente resistente à penicilina.

  2. Intraperitoneal inoculation of Haemophilus influenzae local isolates in BALB/c mice model in the presence and absence of virulence enhancement agents

    Directory of Open Access Journals (Sweden)

    N Mojgani

    2013-01-01

    Full Text Available Purpose:Haemophilus influenzae (Hi, predominantly type b accounts for approximately 4% of cases of community-acquired and nosocomial meningitis, in adults. The objective of this study was to evaluate the pathogenicity of local Hi isolates (type b, f and non-typable in BALB/c mice in the presence of virulence enhancement agents. Materials and Methods: Three different concentrations of the Hi isolates were inoculated intraperitoneally in BALB/c mice in the presence of 2% hemoglobin and 4% mucin as virulence enhancing agents (VEA. The ability of the isolates to produce bacteremia, the percent survival and lethal dose (LD 50 were recorded in different challenge groups. Results: The 3 Haemophilus influenzae type b (Hib isolates used in study were able to show virulence in BALB/c mice model only in the presence of VEA and their LD 50 decreased significantly when 2% hemoglobin and 4% mucin were used. All survived animals showed bacteremia within 4 h of inoculation which was cleared within 18 h. Significant differences ( P < 0.01 in the virulence and survival percentage of Hib challenge groups were observed based on their dose of inoculation and VEA. None of the isolates were able to induce infection in the absence of VEA. Non-type b isolates failed to produce disease in the mice models even at the highest inoculated dose (10 8 cfu and in the presence of VEA. Conclusions: BALB/c mice appeared suitable for evaluating the virulence of Hib strains, and 2% hemoglobin with 4% mucin an appropriate concentration for inducing infection in this animal model.

  3. Antimicrobial susceptibility of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis isolated from community-acquired respiratory tract infections in China: Results from the CARTIPS Antimicrobial Surveillance Program.

    Science.gov (United States)

    Zhang, Yawei; Zhang, Feifei; Wang, Hui; Zhao, Chunjiang; Wang, Zhanwei; Cao, Bin; Du, Yan; Feng, Xianju; Hu, Yunjian; Hu, Bijie; Ji, Ping; Liu, Zhiyong; Liu, Yong; Liao, Wanzhen; Lu, Juan; Sun, Hongli; Wang, Zhongxin; Xu, Xiuli; Xu, Xuesong; Yang, Qing; Yu, Yunsong; Zhang, Rong; Zhuo, Chao

    2016-06-01

    This study investigated the antimicrobial susceptibilities of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis isolates causing adult community-acquired respiratory tract infections (CARTIs) in China. A multicentre resistance surveillance study (CARTIPS) investigating 1046 clinical isolates from 19 hospitals in China was conducted from 2013 to 2014. Based on the minimum inhibitory concentration (MIC) breakpoints of oral penicillin, the percentages of penicillin-resistant, penicillin-intermediate and penicillin-susceptible S. pneumoniae were 44.1%, 13.7%, and 42.2%, respectively. The rates of penicillin-non-susceptible S. pneumoniae ranged from 27.9% to 72.2% in different cities, with the highest rate in Nanchang. Macrolides, including azithromycin, clarithromycin and erythromycin, showed the lowest activities against S. pneumoniae isolates, with resistance rates of 90.5%, 92.2% and 93.0%, respectively. However, 98% of these strains were susceptible to levofloxacin and moxifloxacin. For H. influenzae isolates, most of the antimicrobials agents exhibited good activities. However, ampicillin and trimethoprim/sulfamethoxazole showed relatively lower activity against H. influenzae, with resistance rates of 35.0% and 54.4%, respectively. β-lactamase production rates amongst H. influenzae and M. catarrhalis were 31.0% and 87.1%, respectively. In addition, a total of 15 β-lactamase-negative ampicillin-resistant (BLNAR) strains identified in this study were resistant to ampicillin, amoxicillin/clavulanic acid, cefaclor and cefuroxime. Most of the antimicrobial agents showed excellent activity against M. catarrhalis, with susceptibility rates of >90%. The results from the current study confirmed the regional variations in antimicrobial susceptibility of major CARTI pathogens and provided some choices for the treatment of these organisms. Continuous national surveillance of the epidemiology of CARTIs is strongly warranted in China.

  4. Efficacy of Solithromycin (CEM-101) for Experimental Otitis Media Caused by Nontypeable Haemophilus influenzae and Streptococcus pneumoniae.

    Science.gov (United States)

    Figueira, M; Fernandes, P; Pelton, S I

    2016-09-01

    Solithromycin (CEM-101) is a "fourth-generation" macrolide, as it has three binding site and is acid stable. The three binding sites confer activity against bacteria resistant to the older macrolides and ketolides, including multidrug-resistant Streptococcus pneumoniae and nontypeable Haemophilus influenzae (NTHi). The objective of this study was to evaluate solithromycin pharmacokinetics (PK), middle ear fluid (MEF) concentrations, and microbiologic efficacy in a chinchilla model of experimental otitis media (EOM) due to strains of S. pneumoniae or NTHi. Plasma PK (maximum concentration of drug in serum [Cmax] and area under the concentration-time curve from 0 to 24 h [AUC0-24]) and middle ear fluid (MEF) concentrations were determined. Isolates with specified antimicrobial susceptibility patterns were inoculated directly into the middle ear (ME). Plasma and MEF were collected for PK and MEF cultures performed to determine efficacy. Solithromycin administered at 150 mg/kg of body weight/day resulted in Cmax and AUC0-24 values of 2.2 μg/ml and 27.4 μg · h/ml in plasma and 1.7 μg/ml and 28.2 μg · h/ml in extracellular MEF on day 1. By day 3, Cmax and AUC0-24 values had increased to 4.5 μg/ml and 54 μg · h/ml in plasma and 4.8 μg/ml and 98.6 μg · h/ml in extracellular MEF. For NTHi EOM, three isolates with MIC/minimal bactericidal concentration (MBC) ratios of 0.5/1 μg/ml (isolate BCH1), 2/2 μg/ml (isolate BMC1247C), and 4/4 μg/ml (isolate BMC1213C) were selected. The MEF of >85% of animals infected with BCH1 and BMC1247C was sterilized. For NTHi BMC1213, >85% of MEF cultures remained positive. For S. pneumoniae EOM, 3 isolates with MIC/MBC ratios of 0.06/0.125 μg/ml (S. pneumoniae 331), 0.125/1 μg/ml (S. pneumoniae CP-645 [MLSB phenotype]), and 0.5/2 μg/ml (CP-712 [mefA subclass mefA resistance]) were selected. Solithromycin sterilized MEF in 100% of animals infected with S. pneumoniae 331 and S. pneumoniae CP-645. ME infection persisted in 60% of

  5. Distribution and drug resistance analysis of Streptococcus pneumonia and Haemophilus influenzae%肺炎链球菌和流感嗜血杆菌分布及耐药性分析

    Institute of Scientific and Technical Information of China (English)

    宁静; 韦桂雪

    2013-01-01

    目的:了解临床分离流感嗜血杆菌和肺炎链球菌的分布及耐药情况,为临床合理使用抗菌药物、预防和控制感染提供依据。方法:收集医院2005~2012年各类临床标本分离流感嗜血杆菌92株、肺炎链球菌83株,均经全自动细菌鉴定仪鉴定,用K-B法检测流感嗜血杆菌对常用的15种抗生素及肺炎链球菌对常见的13种抗生素的敏感性,并用头孢硝噻吩纸片法检测流感嗜血杆菌产β-内酰胺酶情况。结果:流感嗜血杆菌和肺炎链球菌主要来源于呼吸道标本,不同季节流感嗜血杆菌的感染率不同,冬春两季为高发季节,肺炎链球菌感染患者的年龄呈双峰分布,以年龄<5岁和>50岁的感染者最多。流感嗜血杆菌对氨苄西林、复方新诺明和头孢呋辛的耐药率较高,产β-内酰胺酶检出率为64.17%。肺炎链球菌对红霉素、氯霉素、四环素、克林霉素耐药性非常严重,其中47株(56.63%)为青霉素不敏感肺炎链球菌(PNSP)。结论:流感嗜血杆菌和肺炎链球菌耐药情况较为严重,对流感嗜血杆菌和肺炎链球菌的耐药性进行严密监测具有重要意义。%Objective:To investigate the characteristics of distribution and drug resistance of Haemophilus influenzae and Streptococcus pneumoniae, provide the basis of using antimicrobial drugs for clinical. Methods:92 clinical isolates of Haemophilus influenzae and 83 isolates of Streptococcus pneumoniae were collected during 2003 to 2012, and then identified by Vitek-2 system. K-B disk method was used to test antimicrobial susceptibility.β-lactamase was detected with nitrocefin disk testing.Results:92 Haemophilus influenzae and 83 Streptococcus pneumoniae of clinical isolates mainly from respiratory, The higher infectious season of Haemophilus influenza was winter and spring. The age of the Streptococcus pneumoniae infection patient is bimodal distribution, mainly50 years old

  6. 临床分离流感嗜血杆菌和副流感嗜血杆菌的耐药性研究%Resistance profiles of Haemophilus influenzae and Haemophilus parainfluenzae isolates in Shanghai

    Institute of Scientific and Technical Information of China (English)

    袁瑾懿; 杨帆; 徐晓刚; 叶信予; 胡付品; 朱德妹

    2009-01-01

    Objective. To understand drug susceptibilities to common antibacterials, resistance mechanism to β-lactams and quinolones and the clonal spread of resistant stains of Haemophilus influenzae (H. influenzae) and Haernophilus parainfluenzae (H. parainfluenzae) isolated from some hospitals in Shanghai. Methods The in vitro antimicrobial susceptibilities to 13 antibacterials, such as ampicillin, of 156 Haemophilus strains collected from 5 hospitals of Shanghai in 2006 were tested by agar dilution method. The β-lactamase production was determined by chromogenic cephalosporin test. TEM and ROB type of β-lactamase genes and quinolone resistance determining regions (QRDR) of ciprofloxacin-resistant strains were detected by polymerase chain reaction (PCR) amplification. The homology of H. influenzae strains were analyzed by enterobacterial repetitive intergenic consensus (ERIC)-PCR. Results The susceptible rate of 109 strains H. influenzae to ampicillin was 74.3%, while those to ampicillin-sulbactam, cephatosporins and fluoroquinolones were all 100.0%. The β-lactamases-producing rates of 109 strains H. influenzae and 47 strains H. parainfluenzae were 25.7% and 19.1% (χ2=0.776,P=0.378), respectively. TEM gene was detected in all β-lactamases-producing strains. Of 109 H. influenzae isolates, only one was resistant to ciprofloxacin, and Ser84Leu mutation was detected in gyrA gene and Gly206Arg mutation in parC gene. The results of ERIC-PCR showed that 106 H. influenzae strains were clustered into 73 groups with similarity level of 85%. Conclusions Clinical isolates of H. influenzae from hospitals in Shanghai remain highly susceptible to common antimicrobial agents except ampicillin. TEM type of β-lactamase production is the main ampicillin-resistant mechanism of the tested stains. The clonal spread of H. influenzae, including ampicillin-resistant strains, is not prevalent.%目的 了解上海部分医院分离的流感和副流感嗜血杆菌对常用抗菌药物的敏

  7. Possible Prevalence and Transmission of Acute Respiratory Tract Infections Caused by Streptococcus pneumoniae and Haemophilus influenzae among the Internally Displaced Persons in Tsunami Disaster Evacuation Camps of Sri Lanka

    OpenAIRE

    Watanabe, Hiroshi; Batuwanthudawe, Ranjith; Thevanesam, Vasanthi; Kaji, Chiharu; Qin, Liang; Nishikiori, Nobuyuki; Saito, Wakana; Saito, Mariko; Watanabe, Kiwao; Oishi, Kazunori; Abeysinghe, Nihal; Kunii, Osamu

    2007-01-01

    Objective The objective of this prospective study was to investigate the status of acute respiratory tract infections caused by Haemophilus influenzae and Streptococcus pneumoniae in tsunami disaster evacuation camps. Methods Nasopharyngeal swabs (NP) of 324 internally displaced persons (IDP) in 3 different tsunami disaster evacuation camps of Sri Lanka were collected between March 18th and 20th, 2005, and analyzed for MIC, β-lactamase production, serotypes, PCR and pulsed-field gel electroph...

  8. Safety and Immunogenicity of a Fully Liquid Vaccine Containing Five-Component Pertussis-Diphtheria-Tetanus-Inactivated Poliomyelitis-Haemophilus influenzae Type B Conjugate Vaccines Administered at Two, Four, Six and 18 Months of Age

    Directory of Open Access Journals (Sweden)

    Ronald Gold

    2007-01-01

    Full Text Available OBJECTIVE: The safety, immunogenicity and lot consistency of a fully liquid, five-component acellular pertussis combination vaccine, comprised of diphteria, tetanus and acellular pertussis, inactivated polio vaccine, Haemophilus influenzae type b (DTaP-IPV-Hib [Pediacel, sanofi pasteur, Canada] were assessed and compared with that of Hib vaccine reconstituted with the five-component acellular pertussis combination vaccine (DTaP-IPV//Hib, Pentacel [sanofi pasteur, Canada].

  9. Safety and Immunogenicity of a Fully Liquid Vaccine Containing Five-Component Pertussis-Diphtheria-Tetanus-Inactivated Poliomyelitis-Haemophilus influenzae Type B Conjugate Vaccines Administered at Two, Four, Six and 18 Months of Age

    OpenAIRE

    Ronald Gold; Luis Barreto; Santiago Ferro; John Thippawong; Roland Guasparini; William Meekison; Margaret Russell; Elaine Mills; Dana Harrison; Pierre Lavigne

    2007-01-01

    OBJECTIVE: The safety, immunogenicity and lot consistency of a fully liquid, five-component acellular pertussis combination vaccine, comprised of diphteria, tetanus and acellular pertussis, inactivated polio vaccine, Haemophilus influenzae type b (DTaP-IPV-Hib [Pediacel, sanofi pasteur, Canada]) were assessed and compared with that of Hib vaccine reconstituted with the five-component acellular pertussis combination vaccine (DTaP-IPV//Hib, Pentacel [sanofi pasteur, Canada]).METHODS: Infants we...

  10. Selection for phase variation of LOS biosynthetic genes frequently occurs in progression of non-typeable Haemophilus influenzae infection from the nasopharynx to the middle ear of human patients.

    Directory of Open Access Journals (Sweden)

    Kate L Fox

    Full Text Available Surface structures in Haemophilus influenzae are subject to rapid ON/OFF switching of expression, a process termed phase variation. We analyse tetranucleotide repeats controlling phase variation in lipo-oligosaccharide (LOS genes of H. influenzae in paired isolates from both the nasopharynx and middle ears of paediatric patients with chronic or recurrent otitis media. A change in expression of at least one of the seven phase variable LOS biosynthesis genes was seen in 12 of the 21 strain pairs. Several strains showed switching of expression in multiple LOS genes, consistent with a key role for phase variable LOS biosynthetic genes in human infection.

  11. Nasopharyngeal colonization by Haemophilus influenzae in children attending day-care centers, in Ribeirão Preto, State of São Paulo, Brazil Colonização da nasofaringe por Haemophilus influenzae em crianças que freqüentam creches em Ribeirão Preto, Estado de São Paulo, Brasil

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    Maria E.N. Bonifácio da Silva

    2006-03-01

    Full Text Available Nasopharyngeal carriage of Haemophilus influenzae (Hi was studied in 114 healthy children A colonização da nasofaringe por Haemophilus influenzae (Hi foi estudada em 114 crianças saudáveis com menos de 3 anos de idade e que freqüentam creches (day-care centers DCC em Ribeirão Preto, estado de São Paulo, Brasil. Para cada uma das cepas isoladas foram determinados o biótipo, o sorotipo (por antisoro especifico e PCR e a sensibilidade a 14 antibióticos. A freqüência de colonização por Hi foi de 72,0%. As cepas isoladas foram identificadas como pertencentes aos biótipos II (36,5%, I (21,5%, V (18,2% e III (16,1%. A freqüência encontrada de cepas encapsuladas foi de 3,2% para o tipo f, 1,0% para o tipo b, 1,0% para o tipo d e 1,0% para o tipo e. A resistência para trimetoprim-sulfametoxazole e ampicilina foi de 46,2% e 10,7% respectivamente. Resistência múltipla foi encontrada em 14 (15,0% das cepas analisadas. 13,9% das cepas analisadas eram produtoras de beta-lactamase, e não foi recuperada nenhuma cepa beta-lactamase negativa e ampicilina resistente. DCCs são considerados locais de risco, com um alto potencial de disseminação de microrganismos e por isto devem ser continuadamente monitorados com a finalidade de detectar a eliminação da colonização da nasofaringe por cepas H. influenzae tipo b das crianças que freqüentam DCC, ou detectar a sua substituição por outro tipo de cepa.

  12. Genes from plasmid pKM101 in Haemophilus influenzae: separation of functions of mucA and mucB

    Energy Technology Data Exchange (ETDEWEB)

    Balganesh, M.; Setlow, J.K.

    1985-11-01

    Haemophilus influenzae, normally not mutable by UV, became UV mutable with a recombinant plasmid insertion. A 7.8-kilobase-pair (kbp) fragment of the plasmid pKM101 containing the mucA and mucB genes was ligated to the shuttle vector pDM2, and a Rec- strain of H. influenzae was transformed with the ligated mixture. All of the transformants, unlike the parent Rec- strain, were resistant to UV, could carry out postreplication repair and Weigle reactivation, showed greatly increased spontaneous mutation, and contained a plasmid carrying an insert of only 1.2 rather than 7.8 kbp. This plasmid in a umuC mutant strain of Escherichia coli complemented a pKM101 derivative lacking mucA function but with an intact mucB gene, although there was no complementation with a mucA+ mucB- plasmid, suggesting that the newly constructed plasmid coded for the mucA protein; this is in accord with the restriction analysis and hybridization between the plasmid and a probe containing all of the mucA gene but only a small fraction of mucB. When one of the H. influenzae Rec- transformants lost the plasmid, the resistance to UV was retained but the high spontaneous mutation and UV mutability were not. The fact that there was hybridization between the chromosome of the cured strain and a probe containing both muc genes but none when almost no mucB was present suggested that at least part of the mucB gene had been integrated into the Rec- chromosome. Five different postreplication repair-proficient strains became UV mutable and had high spontaneous mutation rates caused by the putative mucA plasmid, indicating that these strains already possessed a chromosomal equivalent of the mucB gene.

  13. Haemophilus influenzae : Caracterización de aislamientos recuperados de enfermedades invasivas en Cuba durante el período 2008-2011

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    Gilda Toraño

    2012-12-01

    Full Text Available Con el objetivo de caracterizar los aislamientos de Haemophilus influenzae, responsables de enfermedades invasivas en Cuba, en la etapa posterior a la vacunación se estudió el total de los recuperados durante el período 2008-2011, remitidos al Instituto "Pedro Kourí" desde diferentes provincias del país. La confirmación de especie y determinación de serotipos se realizó atendiendo al requerimiento de los factores V y X y empleando el método de aglutinación en lámina, respectivamente. Se definieron los biotipos a través de las pruebas de indol, urea y ornitina; se determinó la concentración mínima inhibitoria (CMI mediante la prueba del elipsómetro para la ampicilina, la ceftriaxona, el cloranfenicol y la rifampicina. Para 23 aislamientos se corroboró la identificación como H. influenzae : 21 fueron serotipables y 2 no tipables (HNT. El 90,4% de los serotipables fueron serotipo b y se detectaron dos serotipos a. Nueve aislamientos de H. influenzae b pertenecieron al biotipo I y los aislamientos, serotipo a y HNT, al biotipo II. En una cepa se demostró producción de la enzima ß-lactamasa y resistencia para la ampicilina y el cloranfenicol, con valores de CMI= 8 y 12 µg/mL, respectivamente. Se puso en evidencia que a pesar de la disminución de la incidencia de la enfermedad invasiva por Hib, este serotipo aún constituye el más frecuente y se alerta sobre la necesidad de una vigilancia sostenida que permita detectar fallos vacunales. La susceptibilidad antimicrobiana demostrada para este período pudiera considerarse como un beneficio adicional de la introducción de la vacunación en Cuba.

  14. ENFERMEDAD INVASORA POR HAEMOPHILUS INFLUENZAE ANTES Y DESPUÉS DE LA CAMPAÑA DE VACUNACIÓN EN LA POBLACIÓN INFANTIL DE LA COMUNIDAD VALENCIANA (1996-2000

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    Mercedes Goicoechea Sáez

    2002-01-01

    Full Text Available Fundamento: La introducción de la vacuna conjugada anti Haemophilus influenzae tipo b (Hib en niños ha provocado un llamativo descenso de la incidencia de la enfermedad por H. influenzae. El objetivo de este estudio es analizar las características más relevantes de la enfermedad invasora por H. influenzae en cuanto a la epidemiología, clínica, evolución y estado de vacunación de la población infantil de la Comunidad Valenciana en el periodo 1996-2000. Método: Los datos se recogen de las historias clínicas de los niños menores de 15 años que hayan presentado síntomas y signos clínicos sugestivos de enfermedad invasora con aislamiento de Haemophilus influenzae y/o que cumple con los criterios de definición de caso establecidos, atendidos en todos los hospitales públicos de la Comunidad Valenciana entre 1996 y 2000. La evolución de la incidencia se valoró mediante tasas de incidencia. La clínica y su evolución (secuelas y letalidad mediante la frecuencia y distribución por edad. Resultados: Se registraron un total de 36 casos de enfermedad invasora por Haemophilus influenzae. La tasa de incidencia en niños menores de 15 años pasó de 3,56/105 en 1996 a 1,07/105 en 1997 (coincidiendo con la campaña de vacunación y la posterior inclusión de la vacuna conjugada anti Hib en el Calendario de Vacunaciones Sistemáticas de la Comunidad Valenciana y 0,30/105 en 1998, situación que se sigue manteniendo en los años posteriores. El 53% de los casos se dan en menores de 18 meses. Tanto las secuelas como los fallecimientos se producen en la época anterior a la aplicación rutinaria de la vacuna conjugada. Ningún niño vacunado correctamente falleció. Se registraron 2 casos de H. influenzae tipo no b en niños vacunados. Conclusiones: La incidencia de la infección por Haemophilus influenzae tipo b disminuyó drásticamente desde el inicio de la vacunación sistemática de la población infantil.

  15. Enfermedad invasora por Haemophilus Influenzae antes y después de la campaña de vacunación en la población infantil de la Comunidad Valenciana (1996-2000

    Directory of Open Access Journals (Sweden)

    Goicoechea Sáez Mercedes

    2002-01-01

    Full Text Available Fundamento: La introducción de la vacuna conjugada anti Haemophilus influenzae tipo b (Hib en niños ha provocado un llamativo descenso de la incidencia de la enfermedad por H. influenzae. El objetivo de este estudio es analizar las características más relevantes de la enfermedad invasora por H. influenzae en cuanto a la epidemiología, clínica, evolución y estado de vacunación de la población infantil de la Comunidad Valenciana en el periodo 1996-2000. Método: Los datos se recogen de las historias clínicas de los niños menores de 15 años que hayan presentado síntomas y signos clínicos sugestivos de enfermedad invasora con aislamiento de Haemophilus influenzae y/o que cumple con los criterios de definición de caso establecidos, atendidos en todos los hospitales públicos de la Comunidad Valenciana entre 1996 y 2000. La evolución de la incidencia se valoró mediante tasas de incidencia. La clínica y su evolución (secuelas y letalidad mediante la frecuencia y distribución por edad. Resultados: Se registraron un total de 36 casos de enfermedad invasora por Haemophilus influenzae. La tasa de incidencia en niños menores de 15 años pasó de 3,56/10(5 en 1996 a 1,07/10(5 en 1997 (coincidiendo con la campaña de vacunación y la posterior inclusión de la vacuna conjugada anti Hib en el Calendario de Vacunaciones Sistemáticas de la Comunidad Valenciana y 0,30/10(5 en 1998, situación que se sigue manteniendo en los años posteriores. El 53% de los casos se dan en menores de 18 meses. Tanto las secuelas como los fallecimientos se producen en la época anterior a la aplicación rutinaria de la vacuna conjugada. Ningún niño vacunado correctamente falleció. Se registraron 2 casos de H. influenzae tipo no b en niños vacunados. Conclusiones: La incidencia de la infección por Haemophilus influenzae tipo b disminuyó drásticamente desde el inicio de la vacunación sistemática de la población infantil.

  16. Comparison of Established Diagnostic Methodologies and a Novel Bacterial smpB Real-Time PCR Assay for Specific Detection of Haemophilus influenzae Isolates Associated with Respiratory Tract Infections.

    Science.gov (United States)

    Reddington, Kate; Schwenk, Stefan; Tuite, Nina; Platt, Gareth; Davar, Danesh; Coughlan, Helena; Personne, Yoann; Gant, Vanya; Enne, Virve I; Zumla, Alimuddin; Barry, Thomas

    2015-09-01

    Haemophilus influenzae is a significant causative agent of respiratory tract infections (RTI) worldwide. The development of a rapid H. influenzae diagnostic assay that would allow for the implementation of infection control measures and also improve antimicrobial stewardship for patients is required. A number of nucleic acid diagnostics approaches that detect H. influenzae in RTIs have been described in the literature; however, there are reported specificity and sensitivity limitations for these assays. In this study, a novel real-time PCR diagnostic assay targeting the smpB gene was designed to detect all serogroups of H. influenzae. The assay was validated using a panel of well-characterized Haemophilus spp. Subsequently, 44 Haemophilus clinical isolates were collected, and 36 isolates were identified as H. influenzae using a gold standard methodology that combined the results of matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) and a fucK diagnostic assay. Using the novel smpB diagnostic assay, 100% concordance was observed with the gold standard, demonstrating a sensitivity of 100% (95% confidence interval [CI], 90.26% to 100.00%) and a specificity of 100% (95% CI, 63.06% to 100.00%) when used on clinical isolates. To demonstrate the clinical utility of the diagnostic assay presented, a panel of lower RTI samples (n = 98) were blindly tested with the gold standard and smpB diagnostic assays. The results generated were concordant for 94/98 samples tested, demonstrating a sensitivity of 90.91% (95% CI, 78.33% to 97.47%) and a specificity of 100% (95% CI, 93.40% to 100.00%) for the novel smpB assay when used directly on respiratory specimens. PMID:26109443

  17. Food and Drug Administration Approval for Use of Hiberix as a 3-Dose Primary Haemophilus influenzae Type b (Hib) Vaccination Series.

    Science.gov (United States)

    Briere, Elizabeth C

    2016-01-01

    On January 14, 2016, GlaxoSmithKline Biologicals (Research Triangle Park, North Carolina) received approval from the Food and Drug Administration (FDA) to expand use of Hiberix (Haemophilus b Conjugate Vaccine [Tetanus Toxoid Conjugate]) for a 3-dose infant primary vaccination series at ages 2, 4, and 6 months. Hiberix was first licensed in the United States in August 2009 for use as a booster dose in children aged 15 months through 4 years under the Accelerated Approval Regulations, in response to a Haemophilus influenzae type b (Hib) vaccine shortage that lasted from December 2007 to July 2009 (1). Expanding the age indication to include infants provides another vaccine option in addition to other currently licensed monovalent or combination Hib vaccines recommended for the primary vaccination series.* Hiberix contains 10 μg purified capsular polyribosyl ribitolphosphate (PRP) conjugated to 25 μg tetanus toxoid (PRP-T) and is supplied as a single-dose vial of lyophilized vaccine to be reconstituted with saline diluent. For the 3-dose primary series, a single (0.5 mL) dose should be given by intramuscular injection at ages 2, 4, and 6 months; the first dose may be given as early as age 6 weeks. The recommended catch-up schedule for PRP-T vaccines (http://www.cdc.gov/vaccines/schedules/hcp/child-adolescent.html) should be followed. As previously recommended, a single booster dose should be administered to children aged 15 months through 18 months; to facilitate timely booster vaccination, Hiberix can be administered as early as age 12 months, in accordance with Hib vaccination schedules for routine and catch-up immunization (1-3). PMID:27124887

  18. Food and Drug Administration Approval for Use of Hiberix as a 3-Dose Primary Haemophilus influenzae Type b (Hib) Vaccination Series.

    Science.gov (United States)

    Briere, Elizabeth C

    2016-04-29

    On January 14, 2016, GlaxoSmithKline Biologicals (Research Triangle Park, North Carolina) received approval from the Food and Drug Administration (FDA) to expand use of Hiberix (Haemophilus b Conjugate Vaccine [Tetanus Toxoid Conjugate]) for a 3-dose infant primary vaccination series at ages 2, 4, and 6 months. Hiberix was first licensed in the United States in August 2009 for use as a booster dose in children aged 15 months through 4 years under the Accelerated Approval Regulations, in response to a Haemophilus influenzae type b (Hib) vaccine shortage that lasted from December 2007 to July 2009 (1). Expanding the age indication to include infants provides another vaccine option in addition to other currently licensed monovalent or combination Hib vaccines recommended for the primary vaccination series.* Hiberix contains 10 μg purified capsular polyribosyl ribitolphosphate (PRP) conjugated to 25 μg tetanus toxoid (PRP-T) and is supplied as a single-dose vial of lyophilized vaccine to be reconstituted with saline diluent. For the 3-dose primary series, a single (0.5 mL) dose should be given by intramuscular injection at ages 2, 4, and 6 months; the first dose may be given as early as age 6 weeks. The recommended catch-up schedule for PRP-T vaccines (http://www.cdc.gov/vaccines/schedules/hcp/child-adolescent.html) should be followed. As previously recommended, a single booster dose should be administered to children aged 15 months through 18 months; to facilitate timely booster vaccination, Hiberix can be administered as early as age 12 months, in accordance with Hib vaccination schedules for routine and catch-up immunization (1-3).

  19. Trends in the epidemiology of invasive Haemophilus influenzae disease in Queensland, Australia from 2000 to 2013: what is the impact of an increase in invasive non-typable H. influenzae (NTHi)?

    Science.gov (United States)

    Wan Sai Cheong, J; Smith, H; Heney, C; Robson, J; Schlebusch, S; Fu, J; Nourse, C

    2015-10-01

    Following the introduction of vaccination against Haemophilus influenzae type b (Hib), cases of invasive encapsulated Hib disease have decreased markedly. This study aimed to examine subsequent epidemiological trends in invasive H. influenzae disease in Queensland, Australia and in particular, assess the clinical impact and public health implications of invasive non-typable H. influenzae (NTHi) strains. A multicentre retrospective study was conducted from July 2000 to June 2013. Databases of major laboratories in Queensland including Queensland Forensic and Scientific Services (jurisdictional referral laboratory for isolate typing) were examined to identify cases. Demographic, infection site, Indigenous status, serotype, and mortality data were collected. In total, 737 invasive isolates were identified, of which 586 (79·5%) were serotyped. Hib, NTHi and encapsulated non-b strains, respectively, constituted 12·1%, 69·1% and 18·8% of isolates. The predominant encapsulated non-b strains were f (45·5%) and a (27·3%) serotypes. Of isolates causing meningitis, 48·9% were NTHi, 14·9% Hib, 14·9% Hie, 10·6% Hif, 6·4% Hia and 4·3% were untyped. During the study period, there was an increase in the incidence of invasive NTHi disease (P = 0·007) with seasonal peaks in winter and spring (P 0·001) and Hib (P = 0·039) than non-Indigenous patients. In Queensland, invasive H. influenzae disease is now predominantly encountered in adults and most commonly caused by NTHi strains with demonstrated pathogenicity extending to otherwise young or immunocompetent individuals. Routine public health notification of these strains is recommended and recent available immunization options should be considered. PMID:25762194

  20. Mal de las vacas locas: su influencia en la obtención del antígeno principal de Haemophilus influenzae tipo b

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    Jonatan Hernández-Roche

    2007-01-01

    Full Text Available Se evaluaron los parámetros fundamentales del proceso de obtención del polisacárido capsular de Haemophilus influenzae tipo b tales como: medio de cultivo, tipo de hemina empleada como factor de crecimiento y la expresión de cápsula polisacarídica por diferentes cepas. Los medios de cultivo evaluados fueron: infusión cerebro corazón, Mueller Hinton y una variante de Frantz modificado. Las heminas que fueron evaluadas procedieron de las casas comerciales siguientes: Fluka (origen bovino, Sigma (origen equino, ICN (origen porcino y Calbiochem (origen bovino. Las cepas evaluadas fueron H1am/90, H1am/97, H2am/ 96 y Eagan; y la 170 no tipable como control negativo. La concentración del polisacárido capsular fue determinada mediante el método de Finn, en los tres experimentos realizados. Los resultados demostraron que la variante de Frantz modificado permitió un crecimiento de la bacteria cuatro veces mayor, con una mayor expresión polisacarídica superior con respecto a los demás medios evaluados (p < 0,05. Este medio cumple satisfactoriamente las regulaciones de la OMS. Con la utilización de la hemina de la casa Fluka, se obtuvo un incremento de la expresión polisacarídica de 17,2 % superior a las restantes (p < 0,05; sin embargo, su uso está contraindicado por su origen. El empleo de la heminas de las casas Sigma e ICN arrojaron resultados satisfactorios, lo que permite su utilización en los cultivos de Haemophilus influenzae tipo b. Por otra parte, la cepa H1am/97 evidenció una expresión polisacarídica superior (p < 0,05 entre las evaluadas, lo que demuestra la variabilidad existente por parte de las cepas evaluadas.

  1. 氨苄西林耐药流感嗜血杆菌的基因分型研究%Genotyping of ampicillin-resistant Haemophilus influenzae

    Institute of Scientific and Technical Information of China (English)

    姜敏; 王亚娟; 高薇; 袁林; 沈叙庄; 俞桑洁; 杨永弘

    2005-01-01

    目的了解2000-2003年北京、上海、广州细菌耐药监测项目中,小于5岁呼吸道感染儿童鼻咽部携带氨苄西林(AMP)耐药流感嗜血杆菌(Haemophilus influenzae,Hi)的分子流行病学情况.方法对上述呼吸道感染儿童鼻咽部分离的899株Hi进行AMP敏感性检测,筛选出74株AMP耐药Hi,采用巢式PCR荚膜分型和玻片凝集法,对AMP耐药菌株进行b型Hi(Haemophilus influenzae type b,Hib)检测,并用脉冲电场凝胶电泳(pulsed-field gel electrophoresis,PFGE)和多重PCR两种方法,对AMP耐药菌株进行基因分型.结果 74株AMP耐药Hi中,有2株Hib(占2.7%).PFGE分型74株AMP耐药Hi中有38种基因型,具有克隆传播趋势的有5型,包括41株Hi(占55.4%).其中菌株数最多的为A型,有18株,占24.3%,以2002年上海地区为主.多重PCR分型结果有31型,多重PCR与PFGE分型结果一致率为63.5%.结论北京、上海、广州三地区四年内小于5岁的呼吸道感染儿童鼻咽部携带的AMP耐药Hi有55.4%的菌株存在克隆传播.

  2. 流感嗜血杆菌的研究进展%Advances in Haemophilus influenza Study

    Institute of Scientific and Technical Information of China (English)

    苏映琼

    2007-01-01

    流感嗜血杆菌是嗜血杆菌属(Haemophilus influezae)中对人有致病性的常见细菌,可引起小儿脑膜炎、中耳炎、会厌炎等原发化脓性感染及呼吸道继发感染.现就流感嗜血杆菌病的流行情况、实验室检查及其预防措施等方面的研究进展进行综述.

  3. The iron/heme regulated genes of Haemophilus influenzae: comparative transcriptional profiling as a tool to define the species core modulon

    Directory of Open Access Journals (Sweden)

    Morton Daniel J

    2009-01-01

    Full Text Available Abstract Background Haemophilus influenzae requires heme for aerobic growth and possesses multiple mechanisms to obtain this essential nutrient. Although an understanding of the heme acquisition mechanisms of H. influenzae is emerging, significant gaps in our knowledge remain. Unresolved issues include the identities of all genes exhibiting altered transcription in response to iron and heme availability, the fraction of such genes functioning in iron/heme acquisition, and the heterogeneity of this gene set among clinical isolates. Previously we utilized H. influenzae strain Rd KW20 to demonstrate the utility of transcriptional profiling in defining the genes exhibiting altered transcription in response to environmental iron and heme levels. The current study expands upon those observations by determining the iron/heme modulons of two clinical isolates, the type b isolate 10810 and the nontypeable isolate R2866. These data are used to begin to define the core iron/heme modulon of the species. Results Microarray studies were performed to compare gene expression on transition from iron/heme-restricted to iron/heme-replete conditions for each isolate. Of 1820 ORFs on the array corresponding to R2866 genes, 363 were significantly differentially expressed: 233 were maximally transcribed under iron/heme-replete conditions and 130 under iron/heme-restricted conditions. Of the 1883 ORFs representing genes of strain 10810, 353 were significantly differentially transcribed: 150 were preferentially transcribed under iron/heme-replete conditions and 203 under iron/heme-restricted conditions. Comparison of the data sets indicated that 163 genes exhibited similar regulation in both isolates and that 74 of these exhibited similar patterns of regulation in Rd KW20. These comprise the putative core iron/heme modulon. Conclusion This study provides evidence for a conserved core of H. influenzae genes the transcription of which is altered by the availability of

  4. Rapid discrimination of Haemophilus influenzae, H. parainfluenzae, and H. haemolyticus by fluorescence in situ hybridization (FISH and two matrix-assisted laser-desorption-ionization time-of-flight mass spectrometry (MALDI-TOF-MS platforms.

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    Hagen Frickmann

    Full Text Available BACKGROUND: Due to considerable differences in pathogenicity, Haemophilus influenzae, H. parainfluenzae and H. haemolyticus have to be reliably discriminated in routine diagnostics. Retrospective analyses suggest frequent misidentifications of commensal H. haemolyticus as H. influenzae. In a multi-center approach, we assessed the suitability of fluorescence in situ hybridization (FISH and matrix-assisted laser-desorption-ionization time-of-flight mass-spectrometry (MALDI-TOF-MS for the identification of H. influenzae, H. parainfluenzae and H. haemolyticus to species level. METHODOLOGY: A strain collection of 84 Haemophilus spp. comprising 50 H. influenzae, 25 H. parainfluenzae, 7 H. haemolyticus, and 2 H. parahaemolyticus including 77 clinical isolates was analyzed by FISH with newly designed DNA probes, and two different MALDI-TOF-MS systems (Bruker, Shimadzu with and without prior formic acid extraction. PRINCIPAL FINDINGS: Among the 84 Haemophilus strains analyzed, FISH led to 71 correct results (85%, 13 uninterpretable results (15%, and no misidentifications. Shimadzu MALDI-TOF-MS resulted in 59 correct identifications (70%, 19 uninterpretable results (23%, and 6 misidentifications (7%, using colony material applied directly. Bruker MALDI-TOF-MS with prior formic acid extraction led to 74 correct results (88%, 4 uninterpretable results (5% and 6 misidentifications (7%. The Bruker MALDI-TOF-MS misidentifications could be resolved by the addition of a suitable H. haemolyticus reference spectrum to the system's database. In conclusion, no analyzed diagnostic procedure was free of errors. Diagnostic results have to be interpreted carefully and alternative tests should be applied in case of ambiguous test results on isolates from seriously ill patients.

  5. Simultaneous detection of Neisseria meningitidis, Haemophilus influenzae and Streptococcus sp. by polymerase chain reaction for the diagnosis of bacterial meningits Detecção simultânea da Neisseria meningitidis, Haemophilus influenzae e Streptococcus sp. pela reação em cadeia da polimerase no diagnóstico das meningites bacterianas

    Directory of Open Access Journals (Sweden)

    Luciane Failace

    2005-12-01

    Full Text Available The simultaneous detection of Neisseria meningitidis, Haemophilus influenzae, and Streptococcus sp. was assessed by polymerase chain reaction (PCR for the diagnosis of bacterial meningitis, as well as the applicability of PCR as a routine test. A cohort study was carried out with 182 children (2 months to 12 years of age with suspicion of bacterial meningitis. Routine tests identified the etiologic agent in 65/84 children whose clinical status and laboratory findings suggested the presence of bacterial meningitis. Bacterial meningitis was ruled out in 98 children. In 19 children, the etiologic diagnosis was not possible using standard methods; in 14 of these patients, the etiologic agent was identified by PCR (N. meningitidis=12; H. influenzae=1; Streptococcus sp.=1. The sensitivity of PCR was 88.1%; specificity, 99.0%; positive predictive value, 98.7%; and negative predictive, 90.1%. PCR is a useful complementary diagnostic technique, especially when Gram stain, culture, or antigenic detection are negative or inconclusive.Avaliamos o desempenho da reação em cadeia da polimerase (PCR para detecção simultânea da Neisseria meningitidis, Haemophilus influenzae e Streptococcus sp. no diagnóstico das meningites bacterianas e sua aplicabilidade na rotina diagnóstica. Foi realizado um estudo de coorte com 182 crianças apresentando suspeita de meningite bacteriana. Em 84, havia alterações clínicas e laboratoriais sugestivas de meningite bacteriana. Destas, 65 tiveram o agente etiológico identificado pelos métodos laboratoriais de rotina e 19 ficaram sem diagnóstico etiológico. Em 98 pacientes foi excluído o diagnóstico de meningite bacteriana. Analisando o desempenho da PCR encontramos sensibilidade de 88,1%, especificidade de 99,0% e valores preditivos positivo e negativo de 98,7% e 90,1% respectivamente. Nos 19 pacientes com meningite bacteriana mas sem diagnóstico etiológico a PCR detectou microrganismos em 14, sendo 12 N

  6. 不可分型流感嗜血杆菌候选疫苗的研究进展%RESEARCH PROGRESS ON NONTYPEABLE HAEMOPHILUS INFLUENZAE VACCINE CANDIDATES

    Institute of Scientific and Technical Information of China (English)

    何多姣; 张彦霞; 贾天军; 张玉妥

    2011-01-01

    不可分型流感嗜血杆菌(nontypeable Hae;ophilus influenzae,NTHi)是多种感染性疾病如急性中耳炎,鼻窦炎,结膜炎,慢性支气管炎的常见病原菌,近年来随着NTHi菌株耐药性的增强,NTHi感染已经成为全球一个较严重的健康问题,因此目前亟待研究一种安全有效的疫苗来预防NTHi的感染.多种NTHi的抗原被作为抗NTHi感染的候选疫苗来研究,该文主要针对近年来的一些研究热点如外膜蛋白P6/P5/P2/P26、Haps蛋白、HMW蛋白(high molecular weight protein,HMW protein)等进行综述,为后续疫苗的研制提供一些依据.%Nontypeable Haemophilus influenzae (NTHi) is a commonly causative pathogen for many infectious diseases, such as acute otitis media, sinusitis, conjunctivitis, chronic bronchitis and so on. With a recent increase in antibiotic resistance of NTHi, infection caused by NTHi has been a serious global health problem, so it is high time to develop a safe and effective vaccine to prevent against NTHi infection. Many antigens of NTHi have been evaluated as vaccine candidates for NTHi, this article will present a conclusion of research highlights such as outer membrane protein P6/P5/P2/P26, Haps protein, HMW protein (high molecular weight protein) for the past few years to offer some foundations for the follow-up vaccine studies.

  7. Children with chronic suppurative lung disease have a reduced capacity to synthesize interferon-gamma in vitro in response to non-typeable Haemophilus influenzae.

    Directory of Open Access Journals (Sweden)

    Susan J Pizzutto

    Full Text Available Chronic suppurative lung disease (CSLD is characterized by the presence of a chronic wet or productive cough and recurrent lower respiratory infections. The aim of this study was to identify features of innate, cell-mediated and humoral immunity that may increase susceptibility to respiratory infections in children with CSLD. Because non-typeable Haemophilus influenzae (NTHi is commonly isolated from the airways in CSLD, we examined immune responses to this organism in 80 age-stratified children with CSLD and compared their responses with 51 healthy control children. Cytokines involved in the generation and control of inflammation (IFN-γ, IL-13, IL-5, IL-10 at 72 hours and TNFα, IL-6, IL-10 at 24 hours were measured in peripheral blood mononuclear cells challenged in vitro with live NTHi. We also measured circulating IgG subclass antibodies (IgG1 and IgG4 to two H. influenzae outer membrane proteins, P4 and P6. The most notable finding was that PBMC from children with CSLD produced significantly less IFN-γ in response to NTHi than healthy control children whereas mitogen-induced IFN-γ production was similar in both groups. Overall there were minor differences in innate and humoral immune responses between CSLD and control children. This study demonstrates that children with chronic suppurative lung disease have an altered systemic cell-mediated immune response to NTHi in vitro. This deficient IFN-γ response may contribute to increased susceptibility to NTHi infections and the pathogenesis of CSLD in children.

  8. The concentration of intracellular nickel in Haemophilus influenzae is linked to its surface properties and cell-cell aggregation and biofilm formation.

    Science.gov (United States)

    Ng, Jiaqi; Kidd, Stephen P

    2013-04-01

    Of the known proteins which use nickel as a co-factor, Haemophilus influenzae contains only urease and glyoxalase I (gloA). We have recently reported that this pathogen harbours a unique nickel uptake system (nikKLMQO-nimR). Unusually, the disruption of the nickel uptake system (nikQ or nimR mutants) resulted in cells that aggregated and formed an increased biofilm compared to the wild type cells. Using a gloA mutant strain and urease-specific inhibitor we showed that this phenotype is not due to the loss-of-function of these enzymes. By generating H. influenzae "resting cells" which are enzymatically inactive but maintain their structural integrity we have shown that the cell aggregation in the nikQ/nimR mutants is not due to the loss of enzymatic function. The nikQ mutant was unable to accumulate nickel but the addition of excess nickel did restore intracellular nickel levels and this resulted in the nikQ mutant returning to the wild type "free-living" phenotype; cells with no aggregation and no biofilm formation. We used a range of techniques which showed that the nikQ mutant possesses changes to its cell surface properties. The mutant was more negatively charged than wild type cells as well as being more hydrophobic. Analysis of the outer membrane constituents showed that there were molecular differences. Although the nikQ mutant appears to grow the same as its wild type cell we have shown that there is a change in the "lifestyle" of these nickel limited cells and this induces changes to the surface of the cell to promote cell-cell aggregation and biofilm formation. PMID:23499478

  9. The roles of epithelial cell contact, respiratory bacterial interactions and phosphorylcholine in promoting biofilm formation by Streptococcus pneumoniae and nontypeable Haemophilus influenzae.

    Science.gov (United States)

    Krishnamurthy, Ajay; Kyd, Jennelle

    2014-08-01

    Streptococcus pneumoniae and nontypeable Haemophilus influenzae (NTHi) often share a common niche within the nasopharynx, both associated with infections such as bronchitis and otitis media. This study investigated how the association between NTHi and S. pneumoniae and the host affects their propensity to form biofilms. We investigated a selection of bacterial strain and serotype combinations on biofilm formation, and the effect of contact with respiratory epithelial cells. Measurement of biofilm showed that co-infection with NTHi and S. pneumoniae increased biofilm formation following contact with epithelial cells compared to no contact demonstrating the role of epithelial cells in biofilm formation. Additionally, the influence of phosphorylcholine (ChoP) on biofilm production was investigated using the licD mutant strain of NTHi 2019 and found that ChoP had a role in mixed biofilm formation but was not the only requirement. The study highlights the complex interactions between microbes and the host epithelium during biofilm production, suggesting the importance of understanding why certain strains and serotypes differentially influence biofilm formation. A key contributor to increased biofilm formation was the upregulation of biofilm formation by epithelial cell factors.

  10. Haemophilus influenzae tipo b: situação epidemiológica no Estado de Minas Gerais, Brasil, 1993 a 1997

    Directory of Open Access Journals (Sweden)

    Sybelle de Souza Castro Miranzi

    2003-10-01

    Full Text Available Entre as doenças invasivas causadas pelo Haemophilus influenzae tipo b (Hib, destacam-se, pela freqüência e gravidade, as pneumonias e as meningites. No período de 1993 a 1997, foram notificados, em Minas Gerais, 720 casos de meningites por Hib, sendo a causa mais freqüente de meningite bacteriana em menores de um ano e a segunda causa no total de meningites. Entretanto, estimou-se uma ocorrência total de 1.160 casos considerando as meningites bacterianas não especificadas. O total de casos estimados de doença invasiva por Hib parece justificar a recente inclusão da vacina no esquema básico de imunizações. O alto custo da vacina reforça a necessidade de melhorar a vigilância epidemiológica da meningite, que constitui uma das fragilidades das ações de controle desta doença.

  11. Haemophilus influenzae tipo b: situação epidemiológica no Estado de Minas Gerais, Brasil, 1993 a 1997

    Directory of Open Access Journals (Sweden)

    Miranzi Sybelle de Souza Castro

    2003-01-01

    Full Text Available Entre as doenças invasivas causadas pelo Haemophilus influenzae tipo b (Hib, destacam-se, pela freqüência e gravidade, as pneumonias e as meningites. No período de 1993 a 1997, foram notificados, em Minas Gerais, 720 casos de meningites por Hib, sendo a causa mais freqüente de meningite bacteriana em menores de um ano e a segunda causa no total de meningites. Entretanto, estimou-se uma ocorrência total de 1.160 casos considerando as meningites bacterianas não especificadas. O total de casos estimados de doença invasiva por Hib parece justificar a recente inclusão da vacina no esquema básico de imunizações. O alto custo da vacina reforça a necessidade de melhorar a vigilância epidemiológica da meningite, que constitui uma das fragilidades das ações de controle desta doença.

  12. Patterns of binding of aluminum-containing adjuvants to Haemophilus influenzae type b and meningococcal group C conjugate vaccines and components.

    Science.gov (United States)

    Otto, Robert B D; Burkin, Karena; Amir, Saba Erum; Crane, Dennis T; Bolgiano, Barbara

    2015-09-01

    The basis of Haemophilus influenzae type b (Hib) and Neisseria meningitidis serogroup C (MenC) glycoconjugates binding to aluminum-containing adjuvants was studied. By measuring the amount of polysaccharide and protein in the non-adsorbed supernatant, the adjuvant, aluminum phosphate, AlPO4, was found to be less efficient than aluminum hydroxide, Al(OH)3 at binding to the conjugates, at concentrations relevant to licensed vaccine formulations and when equimolar. At neutral pH, binding of TT conjugates to AlPO4 was facilitated through the carrier protein, with only weak binding of AlPO4 to CRM197 being observed. There was slightly higher binding of either adjuvant to tetanus toxoid conjugates, than to CRM197 conjugates. This was verified in AlPO4 formulations containing DTwP-Hib, where the adsorption of TT-conjugated Hib was higher than CRM197-conjugated Hib. At neutral pH, the anionic Hib and MenC polysaccharides did not appreciably bind to AlPO4, but did bind to Al(OH)3, due to electrostatic interactions. Phosphate ions reduced the binding of the conjugates to the adjuvants. These patterns of adjuvant adsorption can form the basis for future formulation studies with individual and combination vaccines containing saccharide-protein conjugates. PMID:26194164

  13. Nontypeable Haemophilus influenzae-Induced MyD88 Short Expression Is Regulated by Positive IKKβ and CREB Pathways and Negative ERK1/2 Pathway

    Science.gov (United States)

    Andrews, Carla S.; Miyata, Masanori; Susuki-Miyata, Seiko; Lee, Byung-Cheol; Komatsu, Kensei; Li, Jian-Dong

    2015-01-01

    Airway diseases such as asthma and chronic obstructive pulmonary disease (COPD) are characterized by excessive inflammation and are exacerbated by nontypeable Haemophilus influenzae (NTHi). Airway epithelial cells mount the initial innate immune responses to invading pathogens and thus modulate inflammation. While inflammation is necessary to eliminate a pathogen, excessive inflammation can cause damage to the host tissue. Therefore, the inflammatory response must be tightly regulated and deciphering the signaling pathways involved in this response will enhance our understanding of the regulation of the host inflammatory response. NTHi binds to TLR2 and signal propagation requires the adaptor molecule myeloid differentiation factor 88 (MyD88). An alternative spliced form of MyD88 is called MyD88 short (MyD88s) and has been identified in macrophages and embryonic cell lines as a negative regulator of inflammation. However, the role of MyD88s in NTHi-induced inflammation in airway epithelial cells remains unknown. Here we show that NTHi induces MyD88s expression and MyD88s is a negative regulator of inflammation in airway epithelial cells. We further demonstrate that MyD88s is positively regulated by IKKβ and CREB and negatively regulated by ERK1/2 signaling pathways. Taken together these data indicate that airway inflammation is controlled in a negative feedback manner involving MyD88s and suggest that airway epithelial cells are essential to maintain immune homeostasis. PMID:26669856

  14. Haemophilus influenzae type B and its conjugate vaccine%B型流感嗜血杆菌及其疫苗的研究进展

    Institute of Scientific and Technical Information of China (English)

    郑东旖

    2010-01-01

    B型流感嗜血杆菌(Haemophilus influenzae type B,Hib)是引起小儿严重细菌感染的主要致病菌,可导致脑膜炎、肺炎、败血症、会厌炎、蜂窝组织炎、心包炎、脊髓炎等多种侵袭性疾病.WHO数据表明,全球每年约330万5岁以下儿童受到Hib感染,其中38万~50万儿童死于Hib感染性疾病,存活儿童中30%~40%出现严重的并发症与残疾.文章从Hib的感染现状、临床表现、Hib病原学特点以及Hib的传染源、传播途径、易感人群、年龄分布与危险因素等流行病学特点进行概述,同时回顾Hib疫苗的研发历史,并对目前广泛使用的Hib结合疫苗的免疫原性与保护效果、疫苗接种的安全性及中国Hib结合疫苗的应用策略进行综述.

  15. Nontypeable Haemophilus influenzae-Induced MyD88 Short Expression Is Regulated by Positive IKKβ and CREB Pathways and Negative ERK1/2 Pathway.

    Directory of Open Access Journals (Sweden)

    Carla S Andrews

    Full Text Available Airway diseases such as asthma and chronic obstructive pulmonary disease (COPD are characterized by excessive inflammation and are exacerbated by nontypeable Haemophilus influenzae (NTHi. Airway epithelial cells mount the initial innate immune responses to invading pathogens and thus modulate inflammation. While inflammation is necessary to eliminate a pathogen, excessive inflammation can cause damage to the host tissue. Therefore, the inflammatory response must be tightly regulated and deciphering the signaling pathways involved in this response will enhance our understanding of the regulation of the host inflammatory response. NTHi binds to TLR2 and signal propagation requires the adaptor molecule myeloid differentiation factor 88 (MyD88. An alternative spliced form of MyD88 is called MyD88 short (MyD88s and has been identified in macrophages and embryonic cell lines as a negative regulator of inflammation. However, the role of MyD88s in NTHi-induced inflammation in airway epithelial cells remains unknown. Here we show that NTHi induces MyD88s expression and MyD88s is a negative regulator of inflammation in airway epithelial cells. We further demonstrate that MyD88s is positively regulated by IKKβ and CREB and negatively regulated by ERK1/2 signaling pathways. Taken together these data indicate that airway inflammation is controlled in a negative feedback manner involving MyD88s and suggest that airway epithelial cells are essential to maintain immune homeostasis.

  16. Simulating Immune Interference on the Effect of a Bivalent Glycoconjugate Vaccine against Haemophilus influenzae Serotypes “a” and “b”

    Science.gov (United States)

    Konini, Angjelina; Kang, Mingsong; Moghadas, Seyed M.

    2016-01-01

    Objective. We sought to evaluate the immune responses to a bivalent Haemophilus influenzae glycoconjugate vaccine against serotypes “a” (Hia) and “b” (Hib) in the presence of the preexisting immunity to Hib. Methods. We developed a stochastic simulation model of humoral immune response to investigate the antigenic challenge of a bivalent combined glycoconjugate vaccine and a bivalent unimolecular glycoconjugate vaccine. We compared simulation outcomes in the absence of any preexisting immunity with an already primed immune response having specific memory B cells and/or anti-Hib antibodies. Results. The simulation results show that the preexisting immune responses to Hib or carrier protein (CP) may significantly impede the production of anti-Hia antibodies by a unimolecular vaccine. In contrast, the production of anti-Hia antibodies using a combined vaccine is inhibited only in the presence of CP immune responses. Conclusions. Preexisting immunity to Hib and CP may play a critical role in the development of immune responses against Hia or Hib using bivalent combined and unimolecular vaccine formulations. Our results suggest that a bivalent combined glycoconjugate vaccine with a carrier protein not previously used in Hib conjugate vaccines may be an effective formulation for generating immune responses to protect against both Hib and Hia infections. PMID:27366171

  17. IDENTIFICATION AND DRUG RESISTANCE OF THE HAEMOPHILUS INFLUENZA%105株流感嗜血杆菌的分离、鉴定与药敏分析

    Institute of Scientific and Technical Information of China (English)

    彭少华; 李从荣; 李栋; 王婷

    2001-01-01

    目的为提高流感嗜血杆菌(Haemophilus influenza,HIN)的分离率及了解该菌的耐药现状.方法配制富含X和V因子的选择性巧克力培养基,对1 308份痰及咽拭标本进行HIN分离、鉴定,用头孢硝噻吩法测试β-内酰胺酶的产生率,并用KB法进行药敏试验.结果呼吸道标本中流感嗜血杆菌的分离率为8.03%;HIN的β-内酰胺酶产酶率为14.0%.HIN药敏试验对抗菌药物的敏感率依次为:阿齐霉素>头孢呋新>头孢噻肟>环丙沙星>氨苄西林>头孢他啶>氯霉素>复方新诺明.结论制作富含X和V因子的选择性培养基和床边接种标本能提高HIN的分离率.HIN感染目前可选用阿齐霉素和二代头孢类药物进行治疗,临床应关注HIN的耐药性变化.

  18. The HMW1 and HMW2 Adhesins Enhance the Ability of Nontypeable Haemophilus influenzae To Colonize the Upper Respiratory Tract of Rhesus Macaques.

    Science.gov (United States)

    Rempe, Katherine A; Porsch, Eric A; Wilson, Jolaine M; St Geme, Joseph W

    2016-10-01

    Nontypeable Haemophilus influenzae (NTHi) initiates infection by colonizing the upper respiratory tract and is a common cause of localized respiratory tract disease. Previous work has established that the NTHi HMW1 and HMW2 proteins are potent adhesins that mediate efficient in vitro adherence to cultured human respiratory epithelial cells. In this study, we used a rhesus macaque model to assess the contributions of HMW1 and HMW2 to in vivo colonization. In experiments involving inoculation of individual isogenic derivatives of NTHi strain 12, the parent strain expressing both HMW1 and HMW2 and the mutant strains expressing either HMW1 or HMW2 were able to colonize more frequently than the double mutant strain lacking HMW1 and HMW2. In competition experiments, the parent strain efficiently outcompeted the double mutant lacking HMW1 and HMW2. Colonization with strains expressing HMW2 resulted in development of antibody against HMW2 in a number of the animals, demonstrating that colonization can stimulate an antibody response. In conclusion, we have established that the HMW1 and HMW2 adhesins play a major role in facilitating colonization of the upper respiratory tract of rhesus macaques, in some cases associated with stimulation of an immune response.

  19. Simulating Immune Interference on the Effect of a Bivalent Glycoconjugate Vaccine against Haemophilus influenzae Serotypes “a” and “b”

    Directory of Open Access Journals (Sweden)

    Angjelina Konini

    2016-01-01

    Full Text Available Objective. We sought to evaluate the immune responses to a bivalent Haemophilus influenzae glycoconjugate vaccine against serotypes “a” (Hia and “b” (Hib in the presence of the preexisting immunity to Hib. Methods. We developed a stochastic simulation model of humoral immune response to investigate the antigenic challenge of a bivalent combined glycoconjugate vaccine and a bivalent unimolecular glycoconjugate vaccine. We compared simulation outcomes in the absence of any preexisting immunity with an already primed immune response having specific memory B cells and/or anti-Hib antibodies. Results. The simulation results show that the preexisting immune responses to Hib or carrier protein (CP may significantly impede the production of anti-Hia antibodies by a unimolecular vaccine. In contrast, the production of anti-Hia antibodies using a combined vaccine is inhibited only in the presence of CP immune responses. Conclusions. Preexisting immunity to Hib and CP may play a critical role in the development of immune responses against Hia or Hib using bivalent combined and unimolecular vaccine formulations. Our results suggest that a bivalent combined glycoconjugate vaccine with a carrier protein not previously used in Hib conjugate vaccines may be an effective formulation for generating immune responses to protect against both Hib and Hia infections.

  20. Drug resistance of Haemophilus influenzae in pediatrics in China%我国儿科流感嗜血杆菌耐药分析

    Institute of Scientific and Technical Information of China (English)

    俞桑洁

    2010-01-01

    @@ 流感嗜血杆菌(Haemophilus influenzae,Hi)最初由Pfeiffer在1892年描述,40年后,Pittman确定了6种荚膜分型,a~f型无荚膜型,可作为呼吸道正常菌群也可作为呼吸道和侵袭性感染致病菌.国外报告Hib型(Hib)菌株的携带受各种b型配方疫苗使用的影响(例如蛋白结合疫苗、多糖疫苗).Hi主要引起的感染有儿童肺炎、脑膜炎和菌血症,其中主要由b型菌株引起,而成人社区获得性肺炎、急件中耳炎、急性鼻安炎、慢性支气管炎急性发作通常由无荚膜到菌株引起.

  1. Reversions of two proline-requiring auxotrophs of Haemophilus influenzae by N-methyl-N'-nitro-N-nitrosoguanidine and hydrazine

    Energy Technology Data Exchange (ETDEWEB)

    Kimball, R.F.

    1976-01-01

    New mutation detection systems are described for Haemophilus influenzae. They involve two independently isolated proline auxotrophs which appear to be mutants at different sites in a proline locus (proB) that is very closely linked to a locus (thd) for thymidine requirement. One of the mutants, proB1, appears to revert to prototrophy only by mutations at the locus. The other, proB2, reverts both by mutation at the locus and by unlinked suppressors. The latter account for about 90 percent of the reversions induced by MNNG and by HZ. The close linkage of proB to thd was used to distinguish between true revertants and suppressors by a transformation test. A comparison was made between the mutation induction kinetics of the different classes of revertants and mutations to novobiocin resistance with MNNG and HZ. The very different induction kinetics for these two mutagens previously reported for the novobiocin resistance system were also found for the proline systems. There were some differences between the detection systems, however, in the frequency of induced mutation relative to the spontaneous frequency and, in one case, in the form of the induction curve. It is concluded that the major features of the induction curves reflect the amount of damage done to DNA and so are general for all systems, but that there are some features which are locus- or site-specific.

  2. Role of pre-replication and post-replication processes in mutation induction in Haemophilus influenzae by N-methyl-N'-nitro-N-nitrosoguanidine

    Energy Technology Data Exchange (ETDEWEB)

    Kimball, R.F.; Perdue, S.W.; Boling, M.E.

    1978-01-01

    Studies were carried out on the repair and fixation of premutational damage induced in Haemophilus influenzae by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG). The studies employed a temperature-sensitive DNA elongation mutant (dna9) and its combinations with mutants defective in pyrimidine dimer excision (uvr1, uvr2) and in recombination (rec1). The dna9 mutant is shown to be leaky, allowing about 1% of the normal rate of DNA synthesis at the restrictive temperature. Repair of premutational lesions was detected by a decline in mutation frequency with increasing delay in DNA replication in dna9 at the restrictive temperature. This repair is unaffected by the pyrimidine dimer excision system. Mutation fixation was detected by the ability of DNA from treated and then lysed cells to transfer mutants to recipient cells by transformation. Some fixation occurred at the restrictive temperature but much less than at the non-restrictive temperature suggesting that an appreciable minority of the mutations resulted from lesions introduced near the replication fork but that the majority of mutations arise from lesions introduced at some distance from the fork, perhaps randomly. The DNA synthesized immediately after MNNG treatment is of lower molecular weight than normal and returns to normal with time. This return is blocked in the rec1 mutant, suggesting that recombination is involved. The possible role of this process in MNNG mutagenesis is discussed.

  3. Crystal Structure of a Complex of Surfactant Protein D (SP-D) and Haemophilus influenzae Lipopolysaccharide Reveals Shielding of Core Structures in SP-D-Resistant Strains.

    Science.gov (United States)

    Clark, Howard W; Mackay, Rose-Marie; Deadman, Mary E; Hood, Derek W; Madsen, Jens; Moxon, E Richard; Townsend, J Paul; Reid, Kenneth B M; Ahmed, Abdul; Shaw, Amy J; Greenhough, Trevor J; Shrive, Annette K

    2016-05-01

    The carbohydrate recognition domains (CRDs) of lung collectin surfactant protein D (SP-D) recognize sugar patterns on the surface of lung pathogens and promote phagocytosis. Using Haemophilus influenzae Eagan strains expressing well-characterized lipopolysaccharide (LPS) surface structures of various levels of complexity, we show that bacterial recognition and binding by SP-D is inversely related to LPS chain extent and complexity. The crystal structure of a biologically active recombinant trimeric SP-D CRD complexed with a delipidated Eagan 4A LPS suggests that efficient LPS recognition by SP-D requires multiple binding interactions utilizing the three major ligand-binding determinants in the SP-D binding pocket, with Ca-dependent binding of inner-core heptose accompanied by interaction of anhydro-Kdo (4,7-anhydro-3-deoxy-d-manno-oct-2-ulosonic acid) with Arg343 and Asp325. Combined with enzyme-linked immunosorbent assays (ELISAs) and fluorescence-activated cell sorter (FACS) binding analyses, our results show that extended LPS structures previously thought to be targets for collectins are important in shielding the more vulnerable sites in the LPS core, revealing a mechanism by which pathogens with complex LPS extensions efficiently evade a first-line mucosal innate immune defense. The structure also reveals for the first time the dominant form of anhydro-Kdo.

  4. b型流感嗜血杆菌结合疫苗的稳定性评价%Evaluation on stability of Haemophilus influenzae type b conjugate vaccine

    Institute of Scientific and Technical Information of China (English)

    方国良; 马波; 杨丽; 吴秀丽; 李子财; 徐东梅; 钱雯; 黄镇

    2015-01-01

    目的 评价b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)结合疫苗于2~8 ℃存放的稳定性.方法 选取由玉溪沃森生物技术有限公司生产的上市销售的11批Hib结合疫苗,置2~8℃存放,分别于第0、3、6、9、12、18、24和30个月取样,进行鉴别试验、外观检查、多糖含量、pH值、氯化钠含量、高分子结合物含量、多糖分子大小、效力试验、无菌检查、热原检查、异常毒性检查和细菌内毒素检查,评价疫苗的稳定性.结果 Hib结合疫苗于2~8℃放置30个月,各项指标均符合质量标准的要求.结论 Hib结合疫苗于2~8℃存放30个月,稳定性良好.

  5. 海口市流感嗜血杆菌流行病学及耐药性研究%Epidemiology and Drug Resistance of Haemophilus Influenzae in Haikou City

    Institute of Scientific and Technical Information of China (English)

    林翀; 林明冠; 苏应仙

    2012-01-01

    目的:研究海口市呼吸系统疾病患儿流感嗜血杆菌流行病学及耐药性情况,为疾病的预防和治疗提供参考.方法:收集2006-2010年海口市各医院(海南医学院附属医院、海南省人民医院、海口市人民医院、海南省农垦总医院)临床送检的咽拭子、鼻咽分泌物、痰、脑脊液等标本486例,然后分离鉴定流感嗜血杆菌;采用Etest法测定时分离到的95株流感嗜血杆菌进行抗生素药敏试验检测.结果:流感嗜血杆菌总检出率19.55%,其栓出情况在不同地区和不同民族之间存在差异,考虑其原因可能和社会经济条件有关,流感嗜血杆菌对常用抗生素的体外药敏结果依次为:左氧氟沙星、头孢曲松>阿莫西林/克拉维酸、阿奇霉素>氯霉素>头孢克洛>阿莫西林>氨苄西林>磺胺甲基异嗯唑.结论:海口流感嗜血杆菌由于全年气温较高,其发病率无明显季节性,细菌栓出率在不同地区及不同民族之间存在差别.对左氧氟沙星、头孢曲松、阿莫西林/克拉维酸、阿奇霉素、氯霉素的耐药率低,其可作为临床治疗流感嗜血杆菌感染的首选药物,而对氧苄西林和磺胺甲基异嚷唑的耐药率较高,其作为临床治疗流感嗜血杆菌首选药物的传统地位受到了严重挑战.%Objective: To evaluate epidemiology and drug resistance of Haemophilus influenzae in Haikou City in children respiratory diseases for disease prevention and treatment. Methods: Between 2006 to 2010 in Haikou Children's Hospital, 486 cases of respiratory tract infection, to cultivate isolated Haemophilus influenzae on nasopharyngeal secretions throuh Etest method for separation of 95 Haemophilus influenzae to antibiotics sensitivity test. Results: Haemophilus influenzae overall detection rate was 19.55 percent, and there are differences in the different regions and between different ethnic groups which was considered its causes was related to socio

  6. Resistance mechanism of Haemophilus influenzae against β- lactam antibiotics and fluoroquinolone%流感嗜血杆菌对β内酰胺类和氟喹诺酮类药物的耐药机制

    Institute of Scientific and Technical Information of China (English)

    马池; 吕媛

    2012-01-01

    Objective To investigate the susceptibility of Haemophilus influenzae to p - lactam antimicrobial agents and fluoroquinolones and to explore the resistance mechanism. Methods Minimal inhibitory concentrations ( MIC)of clinical isolate Haemophilus influenzaes against p - lactam antimicrobial agents and fluoroquinolones were evaluated by the serial two - fold agar dilution method. The production or presence of P - lacta-mase was studied by the rapid method utilizing ihe chromogenic cephalo-sporin compound nitrocefin in cultures of Haemophilus influenzaes. Then TEM -1 and ROB - 1 type of p - lactamase genes and quinolones - resistant determining regions ( QRDR) of fluoroquinolone resistant strain were detected by polymerase chain reaction ( PCR) amplification and product sequencing. Results The susceptible rate of the 183 strains Haemophilus influenzae to ampicillin was 73.2% . Otherwise, all the strains were all susceptible to ampicillin - sulbactam, cefotaxime and cefepim. Two ciprofloxacin and 3 moxifloxacin resistance strains were detected, and one of them was not susceptible to both ciprofloxacin and moxifloxacin. Of the 183 strains Haemophilus influenzae, 34 isolates (18. 6% ) were determinated as p - lactamase producing strains. One hundred percent of the p - lactainase positive strains gave a positive result with specific primers for the TEM - 1 gene. None strain was detected for the ROB -1 gene with in 34 strains in all the 183 strains. One or more mutations in GyrA and ParC in QRDR were detected in all the fluoroquinolone resistance Haemophilus influenzaes, even the change of ami-no acids in position 84 or (and) 88 of GyrA occurred more frequently. Conclusion Other than ampicillin, cephalospo-rins and fluoroquinolones show potent activities against Haemophilus influenzae. The major mechanism of Haemophilus influenzae resistance to p - lactam antibiotics is mainly the presence of β - lactamase, mainly TEM - 1 type enzyme. ROB -1 gene should be persistently

  7. Biotyping of Haemophilus using API 10S—an epidemiological tool?

    OpenAIRE

    Mehtar, S.; Afshar, S Amini

    1983-01-01

    One hundred and ninety-nine strains of Haemophilus isolates were biotyped by Kilian's method1 and a modified API 10S strip and the results compared. One hundred percent correlation was found between the two systems. The ONPG test proved of value in differentiating between Haemophilus influenzae and Haemophilus parainfluenzae when there was growth factor disc failure.

  8. Optimization of the purification process of Haemophilus influenzae type b polysaccharide%b型流感嗜血杆菌多糖纯化工艺的优化

    Institute of Scientific and Technical Information of China (English)

    王伟; 马雷钧; 沈坚; 陈晓怿; 马相虎

    2014-01-01

    目的 优化b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)多糖纯化工艺,替代传统的苯酚结合乙醇方法.方法 采用十六烷基三甲基溴化铵提取粗糖,以脱氧胆酸钠结合乙醇多级纯化进一步纯化粗糖,通过超滤浓缩和除菌过滤获得纯化的Hib多糖.采用优化的纯化工艺以中试和放大规模各纯化3批Hib多糖,按照《中华人民共和国药典》2010年版三部的要求检测纯化的Hib多糖.结果 中试和放大规模纯化的各3批Hib多糖的相对分子质量分别为621 800、634 400、659 900和597 200、612 100、583 400,均符合规定的标准.中试和放大规模纯化的各3批Hib多糖的细菌内毒素含量分别为2.0、0.5、0.7 EU/μg和4.0、2.0、1.0 EU/μg,均明显低于规定的标准.鉴别试验显示,中试和放大规模纯化的各批Hib多糖均可与标准Hib抗血清形成明显的沉淀线.结论 优化的Hib多糖纯化工艺具有较好的稳定性,且易于工艺放大,可替代传统工艺用于Hib多糖的纯化.%Objective To optimize the purification process of Haemophilus influenzae type b (Hib) polysaccharide for replacing the traditional combination of phenol and ethanol.Methods Crude Hib polysaccharides were extracted with Cetavlon and further purified with the combination of sodium deoxycholate and ethanol multistage purification,and purified polysaccharides were obtained by ultrafiltration and sterilizing filtration.Three batches of purified Hib polysaccharides were prepared with the optimized purification precess in pilot and other 3 batches in amplifying scales.Prepared Hib polysaccharides were detected in accordance with the Chinese Pharmacopoeia 2010 edition (Volume Ⅲ).Results The relative molecular masses of each 3 batches of Hib polysaccharides purified in pilot and amplifying scales were 621 800,634 400,659 900 and 597 200,612 100,583 400,respectively,and all met standard requirement.The contents of bacterial endotoxin of each 3 batches

  9. 57株流感嗜血杆菌的耐药性分析及分布%Resistance analysis and distribution of haemophilus influenzae bacillus

    Institute of Scientific and Technical Information of China (English)

    赵蕾

    2013-01-01

    目的:了解本院急性呼吸道感染患者临床分离出的流感嗜血杆菌的耐药性及菌株分布情况,为临床合理使用抗菌药物提供依据。方法采用法国梅里埃API NH鉴定条进行菌株鉴定,头孢硝噻吩检测菌株产β-内酰胺酶情况,纸片扩散法检测菌株的药敏情况。结果2012年4月至2012年7月本院临床分离的57株流感嗜血杆菌中有38株对氨苄西林耐药,耐药率66.7%;29株β-内酰胺酶阳性,占50.9﹪;对复方新诺明、头孢呋辛、四环素、阿莫西林/克拉维酸和氯霉素的耐药率依次为82.5%(47/57)、22.8%(13/57)、15.8%(9/57)、12.3%(7/57)和8.8%(5/57),对头孢噻肟、美罗培南、左氧氟沙星和阿奇霉素均敏感。57株流感嗜血杆菌中有12株(21.1%)来源10岁以下患儿,4株(7.0%)来源于30~40岁的患者,41株(71.9%)来源于50岁以上的患者。结论流感嗜血杆菌对临床常用抗菌药物的耐药率相差很大,本院引起的呼吸道感染多见于老年和儿童患者,临床上应根据本院的流行特点合理选用抗菌药物。%Objective To study the resistance and distribution of Haemophilus influenzae isolated from patients with acute respiratory infections in our hospital, and provide a basis for rational use of antimicrobial agents. Method French Merieux APINH identification strips were used for strain identiifcation;β-lactamase of strains were detected by cephalosporin nitroceifn;Antibiotic susceptibility of strains were tested through the disk diffusion assay. Results Total of 57 strains of Haemophilus inlfuenzae were isolated during April 2012 to July 2012. There were 38 strains were resistant to ampicillin, with the resistance rate was 66.7%; 29 strains produced β-lactamase, accounting for 50.9%. The resistance rates to cotrimoxazole, cefuroxime, tetracycline, amoxicillin/clavulanic acid and chloramphenicol were 82.5%(47/57), 22.8%(13/57), 15.8%(9/57), 12

  10. Process development of a New Haemophilus influenzae type b conjugate vaccine and the use of mathematical modeling to identify process optimization possibilities.

    Science.gov (United States)

    Hamidi, Ahd; Kreeftenberg, Hans; V D Pol, Leo; Ghimire, Saroj; V D Wielen, Luuk A M; Ottens, Marcel

    2016-05-01

    Vaccination is one of the most successful public health interventions being a cost-effective tool in preventing deaths among young children. The earliest vaccines were developed following empirical methods, creating vaccines by trial and error. New process development tools, for example mathematical modeling, as well as new regulatory initiatives requiring better understanding of both the product and the process are being applied to well-characterized biopharmaceuticals (for example recombinant proteins). The vaccine industry is still running behind in comparison to these industries. A production process for a new Haemophilus influenzae type b (Hib) conjugate vaccine, including related quality control (QC) tests, was developed and transferred to a number of emerging vaccine manufacturers. This contributed to a sustainable global supply of affordable Hib conjugate vaccines, as illustrated by the market launch of the first Hib vaccine based on this technology in 2007 and concomitant price reduction of Hib vaccines. This paper describes the development approach followed for this Hib conjugate vaccine as well as the mathematical modeling tool applied recently in order to indicate options for further improvements of the initial Hib process. The strategy followed during the process development of this Hib conjugate vaccine was a targeted and integrated approach based on prior knowledge and experience with similar products using multi-disciplinary expertise. Mathematical modeling was used to develop a predictive model for the initial Hib process (the 'baseline' model) as well as an 'optimized' model, by proposing a number of process changes which could lead to further reduction in price. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:568-580, 2016.

  11. Immunoinformatics Approach in Designing Epitope-based Vaccine Against Meningitis-inducing Bacteria (Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae Type b)

    Science.gov (United States)

    Zahroh, Hilyatuz; Ma’rup, Ahmad; Tambunan, Usman Sumo Friend; Parikesit, Arli Aditya

    2016-01-01

    Meningitis infection is one of the major threats during Hajj season in Mecca. Meningitis vaccines are available, but their uses are limited in some countries due to religious reasons. Furthermore, they only give protection to certain serogroups, not to all types of meningitis-inducing bacteria. Recently, research on epitope-based vaccines has been developed intensively. Such vaccines have potential advantages over conventional vaccines in that they are safer to use and well responded to the antibody. In this study, we developed epitope-based vaccine candidates against various meningitis-inducing bacteria, including Streptococcus pneumoniae, Neisseria meningitidis, and Haemophilus influenzae type b. The epitopes were selected from their protein of polysaccharide capsule. B-cell epitopes were predicted by using BCPred, while T-cell epitope for major histocompatibility complex (MHC) class I was predicted using PAProC, TAPPred, and Immune Epitope Database. Immune Epitope Database was also used to predict T-cell epitope for MHC class II. Population coverage and molecular docking simulation were predicted against previously generated epitope vaccine candidates. The best candidates for MHC class I- and class II-restricted T-cell epitopes were MQYGDKTTF, MKEQNTLEI, ECTEGEPDY, DLSIVVPIY, YPMAMMWRNASNRAI, TLQMTLLGIVPNLNK, ETSLHHIPGISNYFI, and SLLYILEKNAEMEFD, which showed 80% population coverage. The complexes of class I T-cell epitopes–HLA-C*03:03 and class II T-cell epitopes–HLA-DRB1*11:01 showed better affinity than standards as evaluated from their ΔGbinding value and the binding interaction between epitopes and HLA molecules. These peptide constructs may further be undergone in vitro and in vivo testings for the development of targeted vaccine against meningitis infection. PMID:27812281

  12. Process development of a New Haemophilus influenzae type b conjugate vaccine and the use of mathematical modeling to identify process optimization possibilities.

    Science.gov (United States)

    Hamidi, Ahd; Kreeftenberg, Hans; V D Pol, Leo; Ghimire, Saroj; V D Wielen, Luuk A M; Ottens, Marcel

    2016-05-01

    Vaccination is one of the most successful public health interventions being a cost-effective tool in preventing deaths among young children. The earliest vaccines were developed following empirical methods, creating vaccines by trial and error. New process development tools, for example mathematical modeling, as well as new regulatory initiatives requiring better understanding of both the product and the process are being applied to well-characterized biopharmaceuticals (for example recombinant proteins). The vaccine industry is still running behind in comparison to these industries. A production process for a new Haemophilus influenzae type b (Hib) conjugate vaccine, including related quality control (QC) tests, was developed and transferred to a number of emerging vaccine manufacturers. This contributed to a sustainable global supply of affordable Hib conjugate vaccines, as illustrated by the market launch of the first Hib vaccine based on this technology in 2007 and concomitant price reduction of Hib vaccines. This paper describes the development approach followed for this Hib conjugate vaccine as well as the mathematical modeling tool applied recently in order to indicate options for further improvements of the initial Hib process. The strategy followed during the process development of this Hib conjugate vaccine was a targeted and integrated approach based on prior knowledge and experience with similar products using multi-disciplinary expertise. Mathematical modeling was used to develop a predictive model for the initial Hib process (the 'baseline' model) as well as an 'optimized' model, by proposing a number of process changes which could lead to further reduction in price. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:568-580, 2016. PMID:26821825

  13. Incorporation of phosphorylcholine into the lipooligosaccharide of nontypeable Haemophilus influenzae does not correlate with the level of biofilm formation in vitro.

    Science.gov (United States)

    Puig, Carmen; Marti, Sara; Hermans, Peter W M; de Jonge, Marien I; Ardanuy, Carmen; Liñares, Josefina; Langereis, Jeroen D

    2014-04-01

    Nontypeable Haemophilus influenzae (NTHi) is an opportunistic pathogen that causes otitis media in children and community-acquired pneumonia or exacerbations of chronic obstructive pulmonary disease in adults. A large variety of studies suggest that biofilm formation by NTHi may be an important step in the pathogenesis of this bacterium. The objective of this report was to determine the relationship between the presence of phosphorylcholine in the lipooligosaccharide of NTHi and the level of biofilm formation. The study was performed on 111 NTHi clinical isolates collected from oropharyngeal samples of healthy children, middle ear fluid of children with otitis media, and sputum samples of patients with chronic obstructive pulmonary disease or community-acquired pneumonia. NTHi clinical isolates presented a large variation in the level of biofilm formation in a static assay and phosphorylcholine content. Isolates collected from the oropharynx and middle ear fluid of children tended to have more phosphorylcholine and made denser biofilms than isolates collected from sputum samples of patients with chronic obstructive pulmonary disease or community-acquired pneumonia. No correlation was observed between biofilm formation and the presence of phosphorylcholine in the lipooligosaccharide for either planktonic or biofilm growth. This lack of correlation was confirmed by abrogating phosphorylcholine incorporation into lipooligosaccharide through licA gene deletion, which had strain-specific effects on biofilm formation. Altogether, we present strong evidence to conclude that there is no correlation between biofilm formation in a static assay and the presence of phosphorylcholine in lipooligosaccharide in a large collection of clinical NTHi isolates collected from different groups of patients. PMID:24452688

  14. Recognition of riboflavin and the capsular polysaccharide of Haemophilus influenzae type b by antibodies generated to the haptenic epitope D-ribitol.

    Science.gov (United States)

    Ravi, G; Venkatesh, Yeldur P

    2014-04-01

    D-Ribitol, a five-carbon sugar alcohol, is an important metabolite in the pentose phosphate pathway; it is an integral part of riboflavin (vitamin B2) and cell wall polysaccharides in most Gram-positive and a few Gram-negative bacteria. Antibodies specific to D-ribitol were generated in New Zealand white rabbits by using reductively aminated D-ribose-BSA conjugate as the immunogen. MALDI-TOF and amino group analyses of ribitol-BSA conjugate following 120 h reaction showed ~27-30 mol of ribitol conjugated per mole BSA. The presence of sugar alcohol in the conjugates was also confirmed by an increase in molecular mass and a positive periodic acid-Schiff staining in SDS-PAGE. Caprylic acid precipitation of rabbit serum followed by hapten affinity chromatography on ribitol-KLH-Sepharose CL-6B resulted in pure ribitol-specific antibodies (~45-50 μg/mL). The affinity constant of ribitol antibodies was found to be 2.9 × 10(7) M(-1) by non-competitive ELISA. Ribitol antibodies showed 100% specificity towards ribitol, ~800% cross-reactivity towards riboflavin, 10-15% cross-reactivity with sorbitol, xylitol and mannitol, and 5-7% cross-reactivity with L-arabinitol and meso-erythritol. The specificity of antibody to ribitol was further confirmed by its low cross-reactivity (0.4%) with lumichrome. Antibodies to D-ribitol recognized the purified capsular polysaccharide of Haemophilus influenzae type b, which could be specifically inhibited by ribitol. In conclusion, antibodies specific to D-ribitol have been generated and characterized, which have potential applications in the detection of free riboflavin and ribitol in biological samples, as well as identification of cell-surface macromolecules containing ribitol. PMID:24643482

  15. Quality of the Haemophilus influenzae type b (Hib) antibody response induced by diphtheria-tetanus-acellular pertussis/Hib combination vaccines.

    Science.gov (United States)

    Denoël, Philippe A; Goldblatt, David; de Vleeschauwer, Isabel; Jacquet, Jeanne-Marie; Pichichero, Michael E; Poolman, Jan T

    2007-10-01

    It has been repeatedly observed that mixing Haemophilus influenzae type b (Hib) conjugate vaccines with acellular pertussis-containing vaccines (diphtheria-tetanus-acellular pertussis [DTPa]) resulted in a reduced magnitude of the anti-polyriboseribitolphosphate antibody response compared to that obtained when Hib vaccines were administered separately and not mixed. Nevertheless, the quality and functionality of the immune responses have been shown to be the same. With the purpose of investigating the quality of the anti-Hib immune responses that are elicited under different vaccination regimens, we report here four primary and booster-based pediatric clinical trials in which Hib vaccine was either mixed with DTPa or diphtheria-tetanus-whole-cell pertussis (DTPw)-based vaccines or was coadministered. Our results show that avidity maturation of the antibodies was lower when primary vaccination involved DTPa mixed with Hib compared to when DTPa and Hib were coadministered. No such difference was observed between mixed and separately administered Hib when associated with DTPa-hepatitis B virus-inactivated poliovirus or DTPw-based vaccines. All different combinations and regimens elicited the same opsonophagocytic and bactericidal activity as well as the same ability to protect in a passive infant rat protection assay. The functional activity of mixed DTPa-based and Hib vaccines was similar to that of mixed DTPw-based/Hib combinations. In conclusion, in vitro and in vivo data as well as postmarketing vaccine effectiveness data attest to the ability of DTPa-based/Hib combination vaccines to effectively prevent Hib-induced disease in children.

  16. Synergistic effect of interleukin 1 alpha on nontypeable Haemophilus influenzae-induced up-regulation of human beta-defensin 2 in middle ear epithelial cells

    Directory of Open Access Journals (Sweden)

    Park Raekil

    2006-01-01

    Full Text Available Abstract Background We recently showed that beta-defensins have antimicrobial activity against nontypeable Haemophilus influenzae (NTHi and that interleukin 1 alpha (IL-1 alpha up-regulates the transcription of beta-defensin 2 (DEFB4 according to new nomenclature of the Human Genome Organization in human middle ear epithelial cells via a Src-dependent Raf-MEK1/2-ERK signaling pathway. Based on these observations, we investigated if human middle ear epithelial cells could release IL-1 alpha upon exposure to a lysate of NTHi and if this cytokine could have a synergistic effect on beta-defensin 2 up-regulation by the bacterial components. Methods The studies described herein were carried out using epithelial cell lines as well as a murine model of acute otitis media (OM. Human cytokine macroarray analysis was performed to detect the released cytokines in response to NTHi exposure. Real time quantitative PCR was done to compare the induction of IL-1 alpha or beta-defensin 2 mRNAs and to identify the signaling pathways involved. Direct activation of the beta-defensin 2 promoter was monitored using a beta-defensin 2 promoter-Luciferase construct. An IL-1 alpha blocking antibody was used to demonstrate the direct involvement of this cytokine on DEFB4 induction. Results Middle ear epithelial cells released IL-1 alpha when stimulated by NTHi components and this cytokine acted in an autocrine/paracrine synergistic manner with NTHi to up-regulate beta-defensin 2. This synergistic effect of IL-1 alpha on NTHi-induced beta-defensin 2 up-regulation appeared to be mediated by the p38 MAP kinase pathway. Conclusion We demonstrate that IL-1 alpha is secreted by middle ear epithelial cells upon exposure to NTHi components and that it can synergistically act with certain of these molecules to up-regulate beta-defensin 2 via the p38 MAP kinase pathway.

  17. IgG responses to Pneumococcal and Haemophilus influenzae protein antigens are not impaired in children with a history of recurrent acute otitis media.

    Directory of Open Access Journals (Sweden)

    Selma P Wiertsema

    Full Text Available BACKGROUND: Vaccines including conserved antigens from Streptococcus pneumoniae and nontypeable Haemophilus influenzae (NTHi have the potential to reduce the burden of acute otitis media. Little is known about the antibody response to such antigens in young children with recurrent acute otitis media, however, it has been suggested antibody production may be impaired in these children. METHODS: We measured serum IgG levels against 4 pneumococcal (PspA1, PspA 2, CbpA and Ply and 3 NTHi (P4, P6 and PD proteins in a cross-sectional study of 172 children under 3 years of age with a history of recurrent acute otitis media (median 7 episodes, requiring ventilation tube insertion and 63 healthy age-matched controls, using a newly developed multiplex bead assay. RESULTS: Children with a history of recurrent acute otitis media had significantly higher geometric mean serum IgG levels against NTHi proteins P4, P6 and PD compared with healthy controls, whereas there was no difference in antibody levels against pneumococcal protein antigens. In both children with and without a history of acute otitis media, antibody levels increased with age and were significantly higher in children colonised with S. pneumoniae or NTHi compared with children that were not colonised. CONCLUSIONS: Proteins from S. pneumoniae and NTHi induce serum IgG in children with a history of acute otitis media. The mechanisms in which proteins induce immunity and potential protection requires further investigation but the dogma of impaired antibody responses in children with recurrent acute otitis media should be reconsidered.

  18. Nontypeable Haemophilus Influenzae Infection Upregulates the NLRP3 Inflammasome and Leads to Caspase-1-Dependent Secretion of Interleukin-1β - A Possible Pathway of Exacerbations in COPD.

    Directory of Open Access Journals (Sweden)

    Johannes Rotta Detto Loria

    Full Text Available Nontypeable Haemophilus influenzae (NTHi is the most common cause for bacterial exacerbations in chronic obstructive pulmonary disease (COPD. Recent investigations suggest the participation of the inflammasome in the pathomechanism of airway inflammation. The inflammasome is a cytosolic protein complex important for early inflammatory responses, by processing Interleukin-1β (IL-1β to its active form.Since inflammasome activation has been described for a variety of inflammatory diseases, we investigated whether this pathway plays a role in NTHi infection of the airways.A murine macrophage cell line (RAW 264.7, human alveolar macrophages and human lung tissue (HLT were stimulated with viable or non-viable NTHi and/or nigericin, a potassium ionophore. Secreted cytokines were measured with ELISA and participating proteins detected via Western Blot or immunohistochemistry.Western Blot analysis of cells and immunohistochemistry of lung tissue detected the inflammasome key components NLRP3 and caspase-1 after stimulation, leading to a significant induction of IL-1β expression (RAW: control at the lower detection limit vs. NTHi 505±111pg/ml, p<0.01. Inhibition of caspase-1 in human lung tissue led to a significant reduction of IL-1β and IL-18 levels (IL-1β: NTHi 24 h 17423±3198pg/ml vs. NTHi+Z-YVAD-FMK 6961±1751pg/ml, p<0.01.Our data demonstrate the upregulation of the NRLP3-inflammasome during NTHi-induced inflammation in respiratory cells and tissues. Our findings concerning caspase-1 dependent IL-1β release suggest a role for the inflammasome in respiratory tract infections with NTHi which may be relevant for the pathogenesis of bacterial exacerbations in COPD.

  19. Research of Haemophilus influenzae type b in China%中国b型流感嗜血杆菌研究历程

    Institute of Scientific and Technical Information of China (English)

    姜敏; 杨永弘

    2006-01-01

    流感嗜血杆菌(Haemophilus influenzae,Hi),尤其是b型(Hib)是引起儿童细菌性感染的主要病原体之一.过去的60年中,很多国家已广泛开展了对Hib各方面的研究,1980年代开始研制Hib疫苗进行预防相关疾病.1996年WHO将Hib蛋白结合疫苗引入各国扩大的计划免疫,至2004年9月,全球有94个国家(包括美洲、欧洲大部和澳洲)将这种疫苗纳入国家计划免疫,另有15个国家在全球疫苗与免疫联盟(GAVI)资助下使用这种疫苗,占全球所有国家和地区使用的一半.而我国尚未列入其中,原因之一是对Hib感染性疾病的认识还不够充分和深入.回顾我国有关Hib研究的整个历程,查询自40年代起至今中国学者关于Hib在国内外杂志发表的文章,现将主要内容总结如下.

  20. The impact of residency and urbanicity on Haemophilus influenzae Type b and pneumococcal immunization in Shanghai Children: a Retrospective Cohort Study.

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    Abram L Wagner

    Full Text Available BACKGROUND: Haemophilus influenzae type b (Hib vaccine and pneumococcal conjugate vaccine (PCV are relatively expensive, newly introduced vaccines in China. This study evaluates the impact of residency and urbanicity on Hib vaccine and PCV coverage for children aged 2 to 7 years living in Shanghai, China, in August 2012. METHODS: In this exploratory cohort study, a sample of children aged 2 to 7 years, all of whom were eligible to have received the complete series of Hib vaccine and PCV, was obtained from the Shanghai Immunization Program Information System. Three measures of vaccination coverage for Hib vaccine and PCV were examined: dose 1 coverage, series completion, and timeliness of dose 1 vaccination. Multivariable binomial regression was used to estimate the difference in vaccination coverage between locals and the floating population. RESULTS: Dose 1 coverage was 50.9% for Hib vaccine and 11.4% for PCV for the 28,141 abstracted pediatric records. For both vaccines, dose 1 coverage was higher in locals than in the floating population. The disparity in coverage between locals and the floating population was greater in suburban areas than urban areas. Of all children who received dose 1, 79.7% completed the Hib vaccine series, and 91.3% completed the PCV series. Timely dose 1 coverage was 8.2% for Hib vaccine and 0.5% for PCV. CONCLUSION: Low vaccination coverage and extremely low levels of timely dose 1 vaccination indicate that current vaccination efforts are inadequate to reduce the burden of Hib and pneumococcal disease among Chinese children, especially infants. Government funding of the Hib vaccine and PCV through the Expanded Program on Immunization would increase uptake and could also ensure that improvement in the timeliness of administration and series completion is targeted for all demographic groups.

  1. Factors affecting the introduction of new vaccines to poor nations: a comparative study of the Haemophilus influenzae type B and hepatitis B vaccines.

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    Aharona Glatman-Freedman

    Full Text Available BACKGROUND: A major effort to introduce new vaccines into poor nations of the world was initiated in recent years with the help of the GAVI alliance. The first vaccines introduced have been the Haemophilus influenzae type B (Hib and the hepatitis B (Hep B vaccines. The introduction of these vaccines during the first phase of GAVI's operations demonstrated considerable variability. We set out to study the factors affecting the introduction of these vaccines. The African Region (AFRO, where new vaccines were introduced to a substantial number of countries during the first phase of GAVI's funding, was selected for this study. METHODOLOGY/PRINCIPAL FINDINGS: GAVI-eligible AFRO countries with a population of 0.5 million or more were included in the study. Countries were analyzed and compared for new vaccine introduction, healthcare indicators, financial indicators related to healthcare and country-level Governance Indicators, using One Way ANOVA, correlation analysis and Qualitative Comparative Analysis (QCA. Introduction of new vaccines into AFRO nations was associated primarily with high country-level Governance Indicator scores. The use of individual Governance Indicator scores, as well as a combined Governance Indicator score we developed, demonstrated similar results. CONCLUSIONS/SIGNIFICANCE: Our study results indicate that good country-level governance is an imperative pre-requisite for the successful early introduction of new vaccines into poor African nations. Enhanced support measures may be required to effectively introduce new vaccines to countries with low governance scores. The combined governance score we developed may thus constitute a useful tool for helping philanthropic organizations make decisions regarding the type of support needed by different countries to achieve success.

  2. b型流行性感冒嗜血杆菌疫苗的现状及展望%Current Status and Future Development of Haemophilus Influenzae Type b Vaccine

    Institute of Scientific and Technical Information of China (English)

    张雁云

    2011-01-01

    Haemophilus influenzae type b(Hib) infection is a global public health issue, mainly causes pneumonia and meningitis in children, and seriously threaten children's health. The introduction of Hib vaccine has opened up a new path on prevention of infectious disease. This paper review the polyribosylribitel phosphate chain, types of carrier protein and their features, immunogenicity and safety of different Hib conjugated vaccine, and the development of combined vaccine. It will be help for providing practical information to immunization program and clinical practice.%b型流行性感冒嗜血杆菌(Haemophilus Influenzae Type B,Hib)感染,是全球性的公共卫生问题,主要引起儿童肺炎和脑膜炎,严重危害儿童的健康.Hib疫苗的诞生为Hib感染性疾病的预防开辟了一条新的道路.现对Hib结合疫苗的多聚核糖基核糖醇磷酸盐糖链、载体蛋白的种类和特点、不同Hib结合疫苗的免疫原性和安全性,以及联合疫苗的发展作了综述,以期为预防接种和临床操作提供参考.

  3. Actividad in vitro 2-bromo-5-(2-bromo-2-nitrovinil-furano frente a Neisseria meningitidis y Haemophilus influenzae (In vitro activity of 2-bromo-5-(2-bromo-2-nitrovinyl-furan in front of Neisseria meningitidis and Haemophilus influenzae

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    Enrique A. Silveira Prado, Oraida González García y Ricardo Medina Marrero.

    2005-07-01

    Full Text Available Se determinó la actividad in vitro del 2-bromo-5-(2-bromo-2-nitrovinil-furano denominado G-1 sintetizado en el Centro de Bioactivos Químicos de la Universidad Central “Marta Abreu” de Las Villas (Cuba, frente a 74 cepas Neisseria meningitidis grupo B y 34 de Haemophilus influenzae. La concentración inhibitoria mínima (CIM se evaluó por el método de las diluciones seriadas en tubos en Caldo Mueller-Hinton suplementado con suero de ternera (N. meningitidis y hemina y NAD (H. influenzae. La concentración bactericida mínima (CBM se determinó por resiembras de 10 µL en placas de Agar Mueller-Hinton con incubación según los requerimientos de los microorganismos de ensayo en atmósfera de CO2 al 6,5%, a partir de todos los tubos donde no se observó crecimiento visible. Además, se calcularon la CIM y CBM 50 y 90% respectivamente y los índices CBM50/CIM50 y CBM90/CIM90. Los valores de la CIM50 y CBM50 (g/mL fueron de 1,9 y 2,5 frente a N. meningitis y de 3,5 y 8,9 frente a H. influenzae. Los valores de la CIM90 y CBM90 (g/mL fueron de 3,5 y 9,3 frente a N. meningitis y de 39,3 y 39,2 frente a H. influenzae. En ambos casos el índice CBM/CIM fluctuó entre 1 y 2,7. Se concluyó, que en las condiciones del ensayo, el G-1 mostró capacidad bacteriostática y bactericida, ligeramente más intensa frente a N. meningitis.

  4. Influenza

    OpenAIRE

    Ferroni, Eliana; Jefferson, Tom

    2011-01-01

    Influenza viruses are constantly altering their antigenic structure, and every year the WHO recommends which strains of influenza should be included in vaccines. During the autumn–winter months, influenza circulates more frequently (influenza seasons), causing a greater proportion of influenza-like illness and sometimes serious seasonal epidemics.The incidence of symptoms depends on the underlying immunity of the population.

  5. Influenza

    OpenAIRE

    Jefferson, Tom

    2009-01-01

    Influenza viruses are constantly altering their antigenic structure, and every year the WHO recommends which strains of influenza should be included in vaccines. During the autumn-winter months, influenza circulates more frequently (influenza seasons), causing a greater proportion of influenza-like illness, and sometimes serious seasonal epidemics.The incidence of infection depends on the underlying immunity of the population.

  6. RecJ, ExoI and RecG are required for genome maintenance but not for generation of genetic diversity by repeat-mediated phase variation in Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Kumar, Gaurav A.; Woodhall, Mark R.; Hood, Derek W.; Moxon, E. Richard [Molecular Infectious Diseases Group, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Headington, Oxford OX3 9DS (United Kingdom); Bayliss, Christopher D. [Department of Genetics, University of Leicester, University Road, Leicester LE1 7RH (United Kingdom)], E-mail: cdb12@le.ac.uk

    2008-04-02

    High levels of genetic diversity are generated in Haemophilus influenzae populations through DNA repeat-mediated phase variation and recombination with DNA fragments acquired by uptake from the external milieu. Conversely, multiple pathways for maintenance of the genome sequence are encoded in H. influenzae genomes. In Escherichia coli, mutations in single-stranded-DNA exonucleases destabilise tandem DNA repeats whilst inactivation of recG can stabilise repeat tracts. These enzymes also have varying effects on recombination. Deletion mutations were constructed in H. influenzae genes encoding homologs of ExoI, RecJ and RecG whilst ExoVII was refractory to mutation. Inactivation of RecJ and RecG, but not ExoI, increased sensitivity to irradiation with ultraviolet light. An increase in spontaneous mutation rate was not observed in single mutants but only when both RecJ and ExoI were mutated. None of the single- or double-mutations increased or decreased the rates of slippage in tetranucleotide repeat tracts. Furthermore, the exonuclease mutants did not exhibit significant defects in horizontal gene transfer. We conclude that RecJ, ExoI and RecG are required for maintenance of the H. influenzae genome but none of these enzymes influence the generation of genetic diversity through mutations in the tetranucleotide repeat tracts of this species.

  7. 249株流感嗜血杆菌的分离与耐药性分析%Analysis on Isolation and Antibiotic Resistance of 249 Strains of Haemophilus influenzae

    Institute of Scientific and Technical Information of China (English)

    曹敏晖; 曹友德; 李浩; 蔡瑞云

    2011-01-01

    目的 了解流感嗜血杆菌(Hi)的检出状况与耐药情况,指导临床合理用药.方法 统计分析2007-2010年我院Hi的分离与耐药情况.结果 在1052份痰及咽拭子标本中分离出Hi 210株,在242份胆汁标本中分离出Hi 16株,在203份胸腹水标本中分离出Hi 12株,在163份眼、耳拭子标本中分离出Hi 11株.体外药敏结果显示Hi对复方磺胺甲恶哇耐药性最高为63.5%,氨苄西林次之为46.2%,而对阿莫西林/棒酸、利福平、头孢呋辛、头孢噻肟、亚氨培南敏感,耐药率≤2.0%.结论 Hi在呼吸道标本中的检出率最高,眼、耳拭子标本中次之,在胆汁、胸腹水中亦有检出.建议眼、耳拭子及胆汁、胸腹水标本作细菌培养时也应同时接种哥伦比亚血平板(COS)、淋球菌选择性平板(VCA)及嗜血杆菌选择性平板(HAE),Hi感染的治疗应首选复方阿莫西林、头孢二代或三代类抗生素.%Objective To investigate the isolation rate and antibiotic resistance rate of Haemophilus influenzae, and to guide the rational use of antibiotic therapy in clinical practice. Methods The data about the isolation rate and antibiotic resistance rate of Haemophilus influenzae from 2007 to 2010 were analyzed with SPSS11.0 statistical software. Results Totally 210 strains of Haemophilus influenzae were isolated from 1,052 specimens of sputum and pharyngeal secretion, 16 strains isolated from 242 bile specimens, 12 strains isolated from 203 hydrothoraxs specimens, and 11 strains isolated from 163 specimens of eye or ear secretion. In vitro drug- sensitivity test showed that the resistance rate of Haemophilus influenzae to trimethoprime sulfanomicles was the highest (63.5%), followed by ampicillin (46.2%). But the resistance rates to amoxicillin/clavalanic acid, rifampicine, cefuroxime, cefotaxime and imipenem were very low (≤2.0%). Conclusions Heamophilus influenzae could be isolated from the secretions of respiratory track, eye, ear, bile

  8. Comparison of two slide agglutination serotyping methods and PCR-based capsule typing for the characterization of Haemophilus influenzae serotypes Comparação de dois métodos de sorotipagem de cápsula por aglutinação em lamina e o método de PCR para a caracterização de sorotipos de Haemophilus influenzae

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    Maria E.N. Bonifácio da Silva

    2006-03-01

    Full Text Available Ninety-three nasopharyngeal Haemophilus influenzae isolates were serotyped by two slide agglutination methods (SAST 1 and SAST 2 and the results compared with those obtained by capsule type-specific PCR. SAST 1 presented a low correlation with results obtained by PCR (75.2% while SAST 2 showed a better agreement with the molecular technique results (93.5%. These findings suggest that SAST 2 could be an alternative method for adequate detection of H.influenzae type b.Noventa e três isolados nasofaringeanos de H. influenzae foram sorotipados através de 2 métodos de aglutinação em lamina (SAST 1 e SAST 2 e os resultados foram comparados com a sorotipagem por PCR. SAST 1 apresentou uma baixa correlação com os resultados obtidos por PCR (75,2% enquanto que SAST 2 mostrou uma melhor concordância com os resultados da técnica molecular (93,5%. Estes resultados indicam que SAST 2 pode ser um método alternativo para a correta detecção de H. influenzae tipo b.

  9. Virulence phenotypes of low-passage clinical isolates of Nontypeable Haemophilus influenzae assessed using the chinchilla laniger model of otitis media

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    Hogg Justin

    2007-06-01

    Full Text Available Abstract Background The nontypeable Haemophilus influenzae (NTHi are associated with a spectrum of respiratory mucosal infections including: acute otitis media (AOM; chronic otitis media with effusion (COME; otorrhea; locally invasive diseases such as mastoiditis; as well as a range of systemic disease states, suggesting a wide range of virulence phenotypes. Genomic studies have demonstrated that each clinical strain contains a unique genic distribution from a population-based supragenome, the distributed genome hypothesis. These diverse clinical and genotypic findings suggest that each NTHi strain possesses a unique set of virulence factors that contributes to the course of the disease. Results The local and systemic virulence patterns of ten genomically characterized low-passage clinical NTHi strains (PittAA – PittJJ obtained from children with COME or otorrhea were stratified using the chinchilla model of otitis media (OM. Each isolate was used to bilaterally inoculate six animals and thereafter clinical assessments were carried out daily for 8 days by blinded observers. There was no statistical difference in the time it took for any of the 10 NTHi strains to induce otologic (local disease with respect to any or all of the other strains, however the differences in time to maximal local disease and the severity of local disease were both significant between the strains. Parameters of systemic disease indicated that the strains were not all equivalent: time to development of the systemic disease, maximal systemic scores and mortality were all statistically different among the strains. PittGG induced 100% mortality while PittBB, PittCC, and PittEE produced no mortality. Overall Pitt GG, PittII, and Pitt FF produced the most rapid and most severe local and systemic disease. A post hoc determination of the clinical origins of the 10 NTHi strains revealed that these three strains were of otorrheic origin, whereas the other 7 were from patients

  10. 健康幼儿流感嗜血杆菌携带状况初查%A screening on the carrier of Haemophilus influenzae in healthy children

    Institute of Scientific and Technical Information of China (English)

    王复甦; 徐丹戈; 徐宝祥; 倪志敏; 方叶珍; 叶菊莲

    2009-01-01

    目的 以杭州市江干区为试点开展婴幼儿流感嗜血杆菌(Haemophilus influenzae,Hi)携带状况的调查工作,为获得浙江省Hi携带状况的系统资料积累经验和数据,同时也为防治Hi相关疾病提供资料.方法 分6月龄至1周岁、1~2周岁、2~5周岁3个年龄组采集婴幼儿咽拭子,进行Hi的分离与鉴定.对Hi分离株进行血清分型和生化分型,并分析对12种抗生素的敏感性试验和青霉素酶产生试验.结果 在101份健康幼儿咽拭标本中,分离到21株Hi,Hi总带菌率20.8%;其中Hib及Hia(f)血清型各1株,带菌率均为1.0%;其他19株为不可分型株(NTHi),带菌率18.8%.咽部分离的Hi对12种临床常用抗生素耐药程度以复方新诺明最高(耐药率为57.1%),其次分别为四环素(14.3%)、头孢克洛(9.5%)和氨苄西林(4.8%).对阿莫西林一克拉维酸、环丙沙星、头孢噻肟、头孢呋肟钠、头孢曲松、阿奇霉素、利福平、氯霉素100%敏感;21株Hi仅1株NTHi产青霉素酶,产酶率为4.8%.结论 健康幼儿Hi带菌以NTHi主,大多小产青霉素酶.

  11. Defining the binding region in factor H to develop a therapeutic factor H-Fc fusion protein against nontypeable Haemophilus influenzae

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    Sandy M. Wong

    2016-04-01

    Full Text Available Nontypeable Haemophilus influenzae (NTHi cause a range of illnesses including otitis media, sinusitis, and exacerbation of chronic obstructive pulmonary disease, infections that contribute to the problem of antibiotic resistance and are themselves often intractable to standard antibiotic treatment regimens. We investigated a strategy to exploit binding of the complement inhibitor Factor H (FH to NTHi as a functional target for an immunotherapeutic containing the NTHi binding domain of FH fused to the Fc domain of IgG1. Chimeric proteins containing the regions that most FH-binding bacteria use to engage human FH, domains 6 and 7 (FH6,7/Fc and/or 18 through 20 (FH18-20/Fc, were evaluated for binding to NTHi. FH6,7/Fc bound strongly to each of seven NTHi clinical isolates tested and efficiently promoted complement-mediated killing by normal human serum. FH18-20/Fc bound weakly to three of the strains but did not promote complement dependent killing. Outer-membrane protein P5 has been implicated in FH binding by NTHi, and FH6,7/Fc binding was greatly diminished in five of seven P5 deficient isogenic mutant strains tested, implicating an alternative FH binding protein in some strains. Binding of FH18-20/Fc was decreased in the P5 mutant of one strain. A murine model was used to evaluate potential therapeutic application of FH6,7/Fc. FH6,7/Fc efficiently promoted binding of C3 to NTHi exposed to mouse serum, and intranasal delivery of FH6,7/Fc resulted in significantly enhanced clearance of NTHi from the lung. Moreover, a P5 deficient mutant was attenuated for survival in the lung model, suggesting that escape mutants lacking P5 would be less likely to replace strains susceptible to FH6,7/Fc. These results provide evidence for the potential utility of FH6,7/Fc as a therapeutic against NTHi lung infection. FH binding is a common property of many respiratory tract pathogens and FH/Fc chimeras may represent promising alternative or adjunctive

  12. Vigilancia epidemiológica centinela de Haemophilus influenzae y Streptococcus pneumoniae en menores de 5 años en el Perú

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    2003-07-01

    Full Text Available Objetivo: Determinar la frecuencia, serotipos y perfil de resistencia antimicrobiana de Haemophilus influenzae (Hi y Streptococcus pneumoniae (Spn en casos de neumonía y meningitis en niños menores de 5 años en el Perú. Materiales y métodos: Entre octubre de 2000 y diciembre de 2001, se implementó la vigilancia centinela de Hi y Spn en dos hospitales de Lima y tres hospitales de otros departamentos (Cusco, Arequipa y Puno. La identificación de serotipos y el estudio de la resistencia antimicrobiana se realizaron en el Instituto Nacional de Salud de Lima, Perú. La susceptibilidad antimicrobiana por la concentración inhibitoria mínima (CIM se realizó por el método de microdilución en placa, siguiendo las pautas del Comité Nacional de Estándares de Laboratorios Clínicos (NCCLS. Resultados: Ingresaron 1 283 casos. Se aislaron 59 cepas (31 de Hi y 28 de Spn, de ellos, 2,3% de las neumonías y 42,5% de las meningitis tuvieron aislamiento bacteriológico. En 10/1210 (0,8% pacientes con neumonía se aisló Hi y Spn en 18/1210 (1,5%. En 21/73 (28,8% de los casos de meningitis se aislaron Hi y Spn en 10/73 (13,7%. Se identificaron los serotipos de Spn: 1, 5, 6A, 11, 14, 19, 19F y 20. Los cepas aislados de Hi fueron del serotipo B. Se identificaron cepas de Spn con resistencia alta a penicilina (3/13, cotrimoxazol (3/13, eritromicina (1/13, cloranfenicol (1/13 y ceftriaxona (1/13; y cepas de Hi altamente resistentes a cotrimoxazol (4/20 y ampicilina (1/20. Conclusiones: Las tasas de aislamiento de Spn y Hi en menores de 5 años fueron bastante bajas. Se hallaron serotipos ya encontrados en Latinoamérica, y se identificaron algunas cepas de Spn y Hi con resistencia a los antibióticos utilizados en los esquemas de tratamiento para neumonía y meningitis. Recomendamos continuar con este sistema de vigilancia, basado en hospitales centinela.

  13. Defining the Binding Region in Factor H to Develop a Therapeutic Factor H-Fc Fusion Protein against Non-Typeable Haemophilus influenzae.

    Science.gov (United States)

    Wong, Sandy M; Shaughnessy, Jutamas; Ram, Sanjay; Akerley, Brian J

    2016-01-01

    Non-typeable Haemophilus influenzae (NTHi) cause a range of illnesses including otitis media, sinusitis, and exacerbation of chronic obstructive pulmonary disease, infections that contribute to the problem of antibiotic resistance and are themselves often intractable to standard antibiotic treatment regimens. We investigated a strategy to exploit binding of the complement inhibitor Factor H (FH) to NTHi as a functional target for an immunotherapeutic containing the NTHi binding domain of FH fused to the Fc domain of IgG1. Chimeric proteins containing the regions that most FH-binding bacteria use to engage human FH, domains 6 and 7 (FH6,7/Fc) and/or 18 through 20 (FH18-20/Fc), were evaluated for binding to NTHi. FH6,7/Fc bound strongly to each of seven NTHi clinical isolates tested and efficiently promoted complement-mediated killing by normal human serum. FH18-20/Fc bound weakly to three of the strains but did not promote complement dependent killing. Outer-membrane protein P5 has been implicated in FH binding by NTHi, and FH6,7/Fc binding was greatly diminished in five of seven P5 deficient isogenic mutant strains tested, implicating an alternative FH binding protein in some strains. Binding of FH18-20/Fc was decreased in the P5 mutant of one strain. A murine model was used to evaluate potential therapeutic application of FH6,7/Fc. FH6,7/Fc efficiently promoted binding of C3 to NTHi exposed to mouse serum, and intranasal delivery of FH6,7/Fc resulted in significantly enhanced clearance of NTHi from the lung. Moreover, a P5 deficient mutant was attenuated for survival in the lung model, suggesting that escape mutants lacking P5 would be less likely to replace strains susceptible to FH6,7/Fc. These results provide evidence for the potential utility of FH6,7/Fc as a therapeutic against NTHi lung infection. FH binding is a common property of many respiratory tract pathogens and FH/Fc chimeras may represent promising alternative or adjunctive therapeutics against such

  14. Two variants among Haemophilus influenzae serotype b strains with distinct bcs4, hcsA and hcsB genes display differences in expression of the polysaccharide capsule

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    Burger Marina

    2008-02-01

    Full Text Available Abstract Background Despite nearly complete vaccine coverage, a small number of fully vaccinated children in the Netherlands have experienced invasive disease caused by Haemophilus influenzae serotype b (Hib. This increase started in 2002, nine years after the introduction of nationwide vaccination in the Netherlands. The capsular polysaccharide of Hib is used as a conjugate vaccine to protect against Hib disease. To evaluate the possible rise of escape variants, explaining the increased number of vaccine failures we analyzed the composition of the capsular genes and the expressed polysaccharide of Dutch Hib strains collected before and after the introduction of Hib vaccination. Results The DNA sequences of the complete capsular gene clusters of 9 Dutch Hib strains were assessed and two variants, designated type I and type II were found. The two variants displayed considerable sequence divergence in the hcsA and hcsB genes, involved in transport of capsular polysaccharide to the cell surface. Application of hcsA type specific PCRs on 670 Hib strains collected from Dutch patients with invasive Hib disease showed that 5% of the strains collected before 1996 were type II. No endogenous type II Hib strains were isolated after 1995 and all type II strains were isolated from 0–4 year old, non-vaccinated children only. Analysis of a worldwide collection of Hib strains from the pre-vaccination era revealed considerable geographic differences in the distribution of the type I and type II strains with up to 73% of type II strains in the USA. NMR analysis of type I and type II capsule polysaccharides did not reveal structural differences. However, type I strains were shown to produce twice as much surface bound capsular polysaccharide. Conclusion Type II strains were only isolated during the pre-vaccination era from young, non-vaccinated individuals and displayed a lower expression of capsular polysaccharide than type I strains. The higher polysaccharide

  15. Crystal structure of YbaK protein from Haemophilus influenzae (HI1434) at 1.8 A resolution: functional implications.

    Science.gov (United States)

    Zhang, H; Huang, K; Li, Z; Banerjei, L; Fisher, K E; Grishin, N V; Eisenstein, E; Herzberg, O

    2000-07-01

    Structural genomics of proteins of unknown function most straightforwardly assists with assignment of biochemical activity when the new structure resembles that of proteins whose functions are known. When a new fold is revealed, the universe of known folds is enriched, and once the function is determined by other means, novel structure-function relationships are established. The previously unannotated protein HI1434 from H. influenzae provides a hybrid example of these two paradigms. It is a member of a microbial protein family, labeled in SwissProt as YbaK and ebsC. The crystal structure at 1.8 A resolution reported here reveals a fold that is only remotely related to the C-lectin fold, in particular to endostatin, and thus is not sufficiently similar to imply that YbaK proteins are saccharide binding proteins. However, a crevice that may accommodate a small ligand is evident. The putative binding site contains only one invariant residue, Lys46, which carries a functional group that could play a role in catalysis, indicating that YbaK is probably not an enzyme. Detailed sequence analysis, including a number of newly sequenced microbial organisms, highlights sequence homology to an insertion domain in prolyl-tRNA synthetases (proRS) from prokaryote, a domain whose function is unknown. A HI1434-based model of the insertion domain shows that it should also contain the putative binding site. Being part of a tRNA synthetases, the insertion domain is likely to be involved in oligonucleotide binding, with possible roles in recognition/discrimination or editing of prolyl-tRNA. By analogy, YbaK may also play a role in nucleotide or oligonucleotide binding, the nature of which is yet to be determined.

  16. Complementary specificity of restriction endonucleases of Diplococcus pneumoniae with respect to DNA methylation. [Haemophilus influenzae, Escherichia coli, Paramecium aurelia

    Energy Technology Data Exchange (ETDEWEB)

    Lacks, S.; Greenberg, B.

    1977-01-01

    Restriction endonucleases Dpn I and Dpn II are produced by two distinct strains of Diplococcus pneumoniae. The two enzymes show complementary specificity with respect to methylation of sites in DNA. From the identity of its cleavage site with that of Mbo I, it appears that Dpn II cleaves at the unmodified sequence 5'-G-A-T-C-3'. Dpn I cleaves at the same sequence when the adenine residue is methylated. Both enzymes produce only double-strand breaks in susceptible DNA. Their susceptibility to Dpn I and not Dpn II shows that essentially all the G-A-T-C sequences are methylated in DNA from the pneumococcal strain that produces Dpn II as well as in DNA from Hemophilus influenzae and Escherichia coli. In the dam-3 mutant of E. coli none of these sequences appear to be methylated. Residual adenine methylation in the dam-3 mutant DNA most likely occurs at different sites. Different but characteristic degrees of methylation at G-A-T-C sites are found in the DNA of bacterial viruses grown in E. coli. DNAs from mammalian cells and viruses are not methylated at this sequence. Mitochondrial DNA from Paramecium aurelia is not methylated, but a small proportion of G-A-T-C sequences in the macronuclear DNA of this eukaryote appear to be methylated. Possible roles of sequence-specific methylation in the accommodation of plasmids, in the replication of DNA, in the regulation of gene function and in the restriction of viral infection are discussed.

  17. Research on PCR determining of type b Haemophilus influenzae strains and immunogenicity of polysaccharide conjugates%几株b型流感嗜血杆菌多糖结合物的免疫原性检定

    Institute of Scientific and Technical Information of China (English)

    袁玉兰; 郭京蓉; 周继唯; 高华

    2012-01-01

    Objective To determine Haemophilus influenzae type b strains in molecular level using PCR,and to study the immunogenicity of capsular polysaccharide conjugates in mice.Methods Extracting genomes using bacterial DNA extract kit from Hoemophilus influenzae type b strains,and PCR for determining the strains through serotyping-specific and capsular genotyping primers respectively.Various capsular polysaccharides conjugated TT respectively,and the conjugates were administered subcutaneously to mice through dilution.After vaccination with two doses,blood samples were collected for the detection of antibody levels to polyribosylribitol phosphate ( PRP),the capsular polysaccharide of Hib.Results All five Haemophilus influenzae type b strains contain type-specific(482 bp) and capsular type (343 bp)DNA fragment through PCR detecting.The DNA fragments were sequenced.BLAST show that these sequences are 100% homology comparing the above strains respectively,and are 99% and 100% homology comparing the GenBank X78559.1 and M19995.1 respectively.The immunogenicity of mice from various capsular polysaccharide conjugates (PRP-TT) was not significantly different by ELISA detecting.Conclusion Through PCR,Haemophilus influenzae type b strain can be determined in molecular level.The immunogenicity of mice from purified capsular polysaccharide conjugates was not different.The study provides a detection means for the features and heredity stability of Haemophilus influenzae type b strain and reference data for the immunogenicity of different polysaccharide conjugates in vaccine research and development and production.%目的 从分子水平检定b型流感嗜血杆菌,研究不同菌株荚膜多糖结合物的免疫原性.方法 提取基因组,通过型特异和荚膜型基因特异引物,利用PCR检定b型流感嗜血杆菌;不同纯化多糖分别与破伤风类毒素(TT)进行耦联结合,结合物原液经稀释免疫小鼠,通过两针免疫,采血进行免疫效力测定.

  18. 阿奇霉素对气管导管内流感嗜血杆菌生物被膜的影响%Effects of azithromycin on the formation of nontypeable haemophilus influenzae biofilm on endotracheal tubes

    Institute of Scientific and Technical Information of China (English)

    王瑛; 王东; 郭治; 石书梅

    2016-01-01

    目的:观察流感嗜血杆菌是否可在气管导管内形成生物被膜并评价阿奇霉素对气管导管内流感嗜血杆菌形成生物被膜的影响。方法选取 Wistar 大鼠120只,分为空白对照组、感染组及给药组;流感嗜血杆菌制备成琼脂小珠,接种于气管插管大鼠,扫描电镜观察气管导管生物被膜形成情况,并通过细菌计数评价疗效。结果接种细菌后第1天扫描电镜可见导管壁表面凹凸不平,外被黏液样物质,并可见细长的菌体被覆其中,给予阿奇霉素治疗后,感染组和给药组均可在气管插管形成生物被膜,但给药组细菌数明显降低。结论流感嗜血杆菌可在气管插管形成生物被膜,阿奇霉素对流感嗜血杆菌气道内导管早期生物被膜有一定抑制作用。%Objective To observe the formation of biofilms on endotracheal tubes ,and to determine the effect of azithromycin on Haemophilus influenzae in biofilms .Methods One hundred and twenty Wistar rats were intubated with a silicone tube ,80 of them(40 rats were model group and 40 were medicine group) were infected by bacteria to establish the animal models of endotra-cheal tube infections the others were ues as control gruop .Formation of biofilms was detected by scanning electron microscope .Via-ble cells counting was carried out to evaluate the effect of azithromycin orally on biofilms on endotracheal tubes .Results Uneven biofilms were formed on the surface of endotracheal tubes ,and slim bodied of bacteria enclosed by a lot of extracellular matrix .Via-ble cells counting was reduced in medicine group compared with model group .Conclusion Haemophilus influenzae can form bio-films on endotracheal tubes .Azithromycin can exert inhibitory effect on haemophilus influenzae biofilms formed on endotracheal tubes .

  19. Optimization of fermentation procedure for Haemophilus influenzae type b%b型流感嗜血杆菌发酵工艺的优化

    Institute of Scientific and Technical Information of China (English)

    李贵凡; 李秀梅; 魏文进; 路福平

    2012-01-01

    Objective To optimize the procedure for fermentation of Haemophilus influenzae type b (Hib) and increase the yield of Hib capsular polysaccharide. Methods The effects of original pH value and glucose concentration on bacterial concentration (A550 value) and polysaccharide yield of medium were investigated,based on which the condition for culture in shake flask was optimized. The effects of DO,constant pH value and glucose supplement on growth of Hib and yield of polysaccharide were investigated,based on which the condition for culture in 20 L fermenter was optimized. Three consecutive batches of Hib were fermented to evaluate the stabib'ty of the optimized procedure. Results The original pH value and glucose concentration for culture in shake-flask were optimized as 7. 2 and 6 g/L respectively. The fermentation procedure was optimized as follows: the DO and pH value were controlled at 20% and 7. 2 respectively,while 200 g/L glucose was supplemented 5 h after inoculation,at a rate of 3 ml/min. Compared with those by the procedure before optimization,the logarithmic growth phase of Hib fermented by the optimized procedure increased from 8 h to about 10 h,while the final concentration (A550 value) of bacteria from 1. 5 to 3. 4,the yield of Hib capsular polysaccharide from 63 to 96 mg/L. The final concentrations (A550 value) of bacteria and yield of Hib capsular polysaccharide of three batches of Hib fermented were 3. 3 ~ 3. 7 and 92 ~ 98 mg/L respectively,indicating high stability of the optimized procedure. Conclusion By the optimized fermentation procedure,the logarithmic growth phase of Hib was prolonged,the growth of bacteria was promoted,and the yield of Hib capsular polysaccharide increased.%目的 优化b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)的发酵工艺,以提高Hib荚膜多糖的产率.方法 通过考察初始pH值和葡萄糖浓度对培养液菌体浓度(A550值)及多糖产率的影响,优化摇瓶培养条件;通过考察溶氧百

  20. Serum Concentrations of Antibodies against Outer Membrane Protein P6, Protein D, and T- and B-Cell Combined Antigenic Epitopes of Nontypeable Haemophilus influenzae in Children and Adults of Different Ages.

    Science.gov (United States)

    Hua, Chun-Zhen; Hu, Wei-Lin; Shang, Shi-Qiang; Li, Jian-Ping; Hong, Li-Quan; Yan, Jie

    2016-02-01

    Nontypeable Haemophilus influenzae (NTHi) is one of the most common etiologies of acute otitis media, rhinosinusitis, and pneumonia. Outer membrane proteins (OMPs) are the main focus in new vaccine development against NTHi, as the H. influenzae type b (Hib) vaccine does not cover noncapsulated NTHi. The OMPs P6 and protein D are the most promising candidate antigens for an NTHi vaccine, and low antibody levels against them in serum may be correlated with infection caused by NTHi. In the current study, we measured the antibody titers against P6, protein D, and their T- and B-cell combined peptide epitopes in healthy individuals of different ages. We found that children B-cell combined antigenic epitopes. Antibody titers increased at ages 1 to 6 months, peaked at 7 months to 3 years, and remained high at 4 to 6 years. The antibody titers started to decrease after 6 years and were the lowest in the 21- to 30-year group. The geometric mean titers (GMTs) of T- and B-cell combined antigenic epitopes in P6 and protein D were positively correlated with those of the protein antigens. Among 12 peptides tested, P6-61, P6-123, and protein D-167 epitopes were better recognized than others in human serum. These findings might contribute to the development of an effective serotype-independent vaccine for H. influenzae. PMID:26677200

  1. Evidence that uv-inducible error-prone repair is absent in Haemophilus influenzae Rd, with a discussion of the relation to error-prone repair of alkylating-agent damage

    Energy Technology Data Exchange (ETDEWEB)

    Kimball, R.F.; Boling, M.E.; Perdue, S.W.

    1977-01-01

    Haemophilus influenzae Rd and its derivatives are mutated either not at all or to only a very small extent by ultraviolet (uv) radiation, x rays, methyl methanesulfonate, and nitrogen mustard, though they are readily mutated by such agents as N-methyl-N'-nitro-N-nitrosoguanidine, ethyl methanesulfonate, and nitrosocarbaryl (NC). In these respects H. influenzae Rd resembles the lexA mutants of Escherichia coli that lack the SOS or reclex uv-inducible error-prone repair system. This similarity is further brought out by the observation that chloramphenicol has little or no effect on post-replication repair after uv irradiation. In E. coli, chloramphenicol has been reported to considerably inhibit post-replication repair in the wild type but not in the lexA mutant. Earlier work has suggested that most or all the mutations induced in H. influenzae by NC result from error-prone repair. Combined treatment with NC and either x rays or uv shows that the NC error-prone repair system does not produce mutations from the lesions induced by these radiations even while it is producing them from its own lesions. It is concluded that the NC error-prone repair system or systems and the reclex error-prone system are different.

  2. b型流感嗜血杆菌结合疫苗接种反应及其处理%Adverse reactions after inoculation of Haemophilus influenzae type b conjugate vaccine and their treatment principles

    Institute of Scientific and Technical Information of China (English)

    樊永贞

    2014-01-01

    Haemophilus influenzae type b (Hib) conjugate vaccine belongs to category Ⅱ vaccine in China.Safety of Hib vaccine is good,and incidence rate of adverse reactions after Hib vaccination is low.Normal reactions after Hib vaccination include local redness,swelling and pain,and mild fever,etc.These symptoms can disappear spontaneously without treatment.Abnormal reactions after Hib vaccination include local blisters and suppuration,systemic allergic rashes and purpura,gastrointestinal reactions,and emotional abnormality,etc.These abnormal reactions can be recovered well after proper treatment.%b型流感嗜血杆菌(Haemophilus in fluenzae type b,Hib)疫苗在中国属于二类疫苗,其安全性良好,接种反应发生率低.Hib疫苗接种后的一般反应包括局部红肿、疼痛,低热等,无需处理可自愈.Hib疫苗接种后的异常反应包括局部水泡和化脓、全身过敏性皮疹和紫癜、消化道反应、情绪异常等,这些异常反应经对症治疗可得到恢复.

  3. Safety and reactogenicity of the combined diphtheria-tetanus-acellular pertussis-inactivated poliovirus-Haemophilus influenzae type b (DTPa-IPV/Hib) vaccine in healthy Vietnamese toddlers: An open-label, phase III study.

    Science.gov (United States)

    Anh, Dang Duc; Van Der Meeren, Olivier; Karkada, Naveen; Assudani, Deepak; Yu, Ta-Wen; Han, Htay Htay

    2016-03-01

    The introduction of combination vaccines plays a significant role in increasing vaccine acceptance and widening vaccine coverage. Primary vaccination against diphtheria, tetanus, pertussis, poliomyelitis and Haemophilus influenza type b (Hib) diseases has been implemented in Vietnam. In this study we evaluated the safety and reactogenicity of combined diphtheria-tetanus-pertussis-inactivated polio (DTPa-IPV)/Hib vaccine when administered as a booster dose in 300 healthy Vietnamese children <2 years of age (mean age: 15.8 months). During the 4-day follow-up period, pain (31.7%) and redness (27.3%) were the most frequent solicited local symptoms. Pain (2%) was also the most frequent grade 3 local symptom. One subject reported 2 serious adverse events that were not causally related to the study vaccine. DTPa-IPV/Hib conjugate vaccine was well tolerated as a booster dose in healthy Vietnamese children aged <2 years. PMID:26337197

  4. Erfðafræðilegur fjölbreytileiki Haemophilus influenzae meðal bera- og sjúkdómsvaldandi stofna á Íslandi 2012

    OpenAIRE

    Jana Birta Björnsdóttir 1989

    2014-01-01

    Haemophilus influenzae (Hi) eru Gram neikvæðir stafir sem finnast í efri loftvegum manna. Hi getur bæði dvalið einkennalaust í nefkoki manna (berar), og orsakað sýkingar, þá sérstaklega þegar ónæmisvarnir hýsilsins eru skertar. Hi eru flokkaðir eftir tilvist fjölsykruhjúps, sex hjúpgerðum hefur verið lýst; a, b, c, d, e og f. Genamengi hjúpaðra Hi sýnir lítinn breytileika innan hverrar hjúpgerðar. Hjúplausir Hi (NTHi) hafa engan fjölsykruhjúp og er genamengi þeirra mjög fjölbreytt. H. haemoly...

  5. Safety and reactogenicity of the combined diphtheria-tetanus-acellular pertussis-inactivated poliovirus-Haemophilus influenzae type b (DTPa-IPV/Hib) vaccine in healthy Vietnamese toddlers: An open-label, phase III study.

    Science.gov (United States)

    Anh, Dang Duc; Van Der Meeren, Olivier; Karkada, Naveen; Assudani, Deepak; Yu, Ta-Wen; Han, Htay Htay

    2016-03-01

    The introduction of combination vaccines plays a significant role in increasing vaccine acceptance and widening vaccine coverage. Primary vaccination against diphtheria, tetanus, pertussis, poliomyelitis and Haemophilus influenza type b (Hib) diseases has been implemented in Vietnam. In this study we evaluated the safety and reactogenicity of combined diphtheria-tetanus-pertussis-inactivated polio (DTPa-IPV)/Hib vaccine when administered as a booster dose in 300 healthy Vietnamese children <2 years of age (mean age: 15.8 months). During the 4-day follow-up period, pain (31.7%) and redness (27.3%) were the most frequent solicited local symptoms. Pain (2%) was also the most frequent grade 3 local symptom. One subject reported 2 serious adverse events that were not causally related to the study vaccine. DTPa-IPV/Hib conjugate vaccine was well tolerated as a booster dose in healthy Vietnamese children aged <2 years.

  6. Actual State of Haemophilus Influenza in Children with Respiratory Tract Infection in Xiaogan%孝感地区儿童呼吸道流感嗜血杆菌感染现状

    Institute of Scientific and Technical Information of China (English)

    胡艳华; 刘东华; 鲁艳

    2011-01-01

    Objective To investigate the infections state and antibiotic resistance of Haemophilus influenza in children with respiratory tract infection in Xiaogan,so as to provide basis for medical treatment and to guide the reasonable use Haemophilus influenza in clinic. Methods The specimens of clinical sputum and swabes samples from children outpatient and be hospitalized with respiratory tract infection were collected,incubated and identified from January 2008 to January 2010 in Xiaogan. Drug susceptibility test was conducted by kirby-bauer method. Nitrocefin slip test was applied to detect β-lactamase. Results Totally 92 Haemophilus influenza strains were isolated during 2008 -2010. The resistance rate to ampicillin.smz + tmp, chloramphenicol, levofloxacin,ciprofloxacin, azithromycin, cefotaxime, cefuroxime, ampicillin/sulbactam were 43. 5% , 57. 6% , 3. 3% , 26% , 4.3% ,2.2% ,4.3% ,8.7% , and 8. 7% respectively. The prevalence of p-lactamase was 32.6%. Conclusion The high drug antibiotic resistance and β-lactamase positive rate of Haemophilus influenza isolated from respiratory tract infection of children by multi-resistance in Xiaogan were serious. It must be paid highly attention by clinicians. The use of the second,third generation of cephalosporin and β-lactamase was the first choice of treatment.%目的 研究孝感地区儿童呼吸道流感嗜血杆菌感染现状及耐药性,为临床抗感染治疗提供依据,以便有效指导临床合理用药.方法 对孝感市中心医院2008年1月-2010年1月儿科门诊及住院患儿呼吸道感染患儿的鼻咽分泌物及深部吸痰,进行细菌培养,菌种鉴定,并用K-B法进行药敏试验,头孢硝噻酚试验检测β-内酰胺酶.结果2年间共分离出Hi 92株,对氨苄西林、复方新诺明、氯霉素的耐药率较高,分别为43.5%、57.6%、26%,对左氧氟沙星、环丙沙星、阿奇霉素、头孢噻肟、头孢呋辛、氨苄西林/舒巴坦的敏感性均较高,耐药率分别为3

  7. Efficacy of pneumococcal nontypable Haemophilus influenzae protein D conjugate vaccine (PHiD-CV in young Latin American children: A double-blind randomized controlled trial.

    Directory of Open Access Journals (Sweden)

    Miguel W Tregnaghi

    2014-06-01

    Full Text Available BACKGROUND: The relationship between pneumococcal conjugate vaccine-induced antibody responses and protection against community-acquired pneumonia (CAP and acute otitis media (AOM is unclear. This study assessed the impact of the ten-valent pneumococcal nontypable Haemophilus influenzae protein D conjugate vaccine (PHiD-CV on these end points. The primary objective was to demonstrate vaccine efficacy (VE in a per-protocol analysis against likely bacterial CAP (B-CAP: radiologically confirmed CAP with alveolar consolidation/pleural effusion on chest X-ray, or non-alveolar infiltrates and C-reactive protein ≥ 40 µg/ml; other protocol-specified outcomes were also assessed. METHODS AND FINDINGS: This phase III double-blind randomized controlled study was conducted between 28 June 2007 and 28 July 2011 in Argentine, Panamanian, and Colombian populations with good access to health care. Approximately 24,000 infants received PHiD-CV or hepatitis control vaccine (hepatitis B for primary vaccination, hepatitis A at booster at 2, 4, 6, and 15-18 mo of age. Interim analysis of the primary end point was planned when 535 first B-CAP episodes, occurring ≥2 wk after dose 3, were identified in the per-protocol cohort. After a mean follow-up of 23 mo (PHiD-CV, n = 10,295; control, n = 10,201, per-protocol VE was 22.0% (95% CI: 7.7, 34.2; one-sided p = 0.002 against B-CAP (conclusive for primary objective and 25.7% (95% CI: 8.4%, 39.6% against World Health Organization-defined consolidated CAP. Intent-to-treat VE was 18.2% (95% CI: 5.5%, 29.1% against B-CAP and 23.4% (95% CI: 8.8%, 35.7% against consolidated CAP. End-of-study per-protocol analyses were performed after a mean follow-up of 28-30 mo for CAP and invasive pneumococcal disease (IPD (PHiD-CV, n = 10,211; control, n = 10,140 and AOM (n = 3,010 and 2,979, respectively. Per-protocol VE was 16.1% (95% CI: -1.1%, 30.4%; one-sided p = 0.032 against clinically confirmed AOM, 67.1% (95% CI: 17.0%, 86

  8. Frequency of apnea, bradycardia, and desaturations following first diphtheria-tetanus-pertussis-inactivated polio-Haemophilus influenzae type B immunization in hospitalized preterm infants

    Directory of Open Access Journals (Sweden)

    Spady Donald W

    2006-06-01

    Full Text Available Abstract Background Adverse cardiorespiratory events including apnea, bradycardia, and desaturations have been described following administration of the first diphtheria-tetanus-pertussis-inactivated polio-Haemophilus influenzae type B (DTP-IPV-Hib immunization to preterm infants. The effect of the recent substitution of acellular pertussis vaccine for whole cell pertussis vaccine on the frequency of these events requires further study. Methods Infants with gestational age of ≤ 32 weeks who received their first DTP-IPV-Hib immunization prior to discharge from two Edmonton Neonatal Intensive Care Units January 1, 1996 to November 30, 2000 were eligible for the study. Each immunized infant was matched by gestational age to one control infant. The number of episodes of apnea, bradycardia, and/or desaturations (ABD and the treatment required for these episodes in the 72 hours prior to and 72 hours post-immunization (for the immunized cohort or at the same post-natal age (for controls was recorded. Results Thirty-four infants who received DTP-IPV-Hib with whole cell pertussis vaccine, 90 infants who received DTP-IPV-Hib with acellular pertussis vaccine, and 124 control infants were entered in the study. Fifty-six immunized infants (45.1% and 36 control infants (29.0% had a resurgence of or increased ABD in the 72 hours post-immunization in the immunized infants and at the same post-natal age in the controls with an adjusted odds ratio for immunized infants of 2.41 (95% CI 1.29,4.51 as compared to control infants. The incidence of an increase in adverse cardiorespiratory events post-immunization was the same in infants receiving whole cell or acellular pertussis vaccine (44.1% versus 45.6%. Eighteen immunized infants (14.5% and 51 control infants (41.1% had a reduction in ABD in the 72 hours post- immunization or at the equivalent postnatal age in controls for an odds ratio of 0.175 (95%CI 0.08, 0.39. The need for therapy of ABD in the immunized

  9. Nontypeable Haemophilus influenzae induces COX-2 and PGE2 expression in lung epithelial cells via activation of p38 MAPK and NF-kappa B

    Directory of Open Access Journals (Sweden)

    Koga Tomoaki

    2008-01-01

    Full Text Available Abstract Background Nontypeable Haemophilus influenzae (NTHi is an important respiratory pathogen implicated as an infectious trigger in chronic obstructive pulmonary disease, but its molecular interaction with human lung epithelial cells remains unclear. Herein, we tested that the hypothesis that NTHi induces the expression of cyclooxygenase (COX-2 and prostaglandin E2 (PGE2 via activation of p38 mitogen-activated protein kinase (MAPK and nuclear factor (NF-kappa B in pulmonary alveolar epithelial cells. Methods Human alveolar epithelial A549 cells were infected with different concentrations of NTHi. The phosphorylation of p38 MAPK was detected by Western blot analysis, the DNA binding activity of NF-kappa B was assessed by electrophoretic mobility shift assay (EMSA, and the expressions of COX-1 and 2 mRNA and PGE2 protein were measured by reverse transcription-polymerase chain reaction (RT-PCR and enzyme linked immunosorbent assay (ELISA, respectively. The roles of Toll-like receptor (TLR 2 and TLR4, well known NTHi recognizing receptor in lung epithelial cell and gram-negative bacteria receptor, respectively, on the NTHi-induced COX-2 expression were investigated in the HEK293 cells overexpressing TLR2 and TLR4 in vitro and in the mouse model of NTHi-induced pneumonia by using TLR2 and TLR4 knock-out mice in vivo. In addition, the role of p38 MAPK and NF-kappa B on the NTHi-induced COX-2 and PGE2 expression was investigated by using their specific chemical inhibitors. Results NTHi induced COX-2 mRNA expression in a dose-dependent manner, but not COX-1 mRNA expression in A549 cells. The enhanced expression of PGE2 by NTHi infection was significantly decreased by pre-treatment of COX-2 specific inhibitor, but not by COX-1 inhibitor. NTHi induced COX-2 expression was mediated by TLR2 in the epithelial cell in vitro and in the lungs of mice in vivo. NTHi induced phosphorylation of p38 MAPK and up-regulated DNA binding activity of NF-kappa B

  10. Optimization of Fermentation Medium for Type b Haemophilus influenzae%b型流感嗜血杆菌发酵培养基的优化

    Institute of Scientific and Technical Information of China (English)

    袁玉兰; 郭京蓉; 高华; 周继唯; 王立新; 林云

    2011-01-01

    目的 分析b型流感嗜血杆菌(Type b Haernophilus influenzae,Hib)不同培养基发酵液中蛋白质、核酸含量及不同培养基添加物对发酵产物的影响,优化Hib发酵培养基.方法 设计不同配方的Hib摇瓶培养基,经摇瓶培养,检测发酵液上清中的蛋白质及核酸含量,选择合适的基础培养基,并在此基础上进行发酵罐培养,检测不同添加物(磷酸盐、钠盐、金属离子等)对Hib多糖复合物收获量、菌体形态及发酵液颜色的影响,并发酵3批,进一步检测多糖复合物中蛋白质和核酸的含量.结果 培养基介质不同,其发酵上清液中的蛋白质和核酸含量也不同,2号培养基(主要成分为酸水解酪素粉和酵母浸粉)发酵液蛋白质和核酸含量较低且稳定;培养基中不同添加物对Hib菌体形态、发酵液颜色及多糖复合物的收获量、质量和性状均产生不同的影响,其中b培养基发酵液菌体形态、多糖复合物收获量及质量均较好;3批发酵培养液中分别约有7%~10%的蛋白质和34%~42%的核酸被沉淀至多糖复合物中.结论 选择2号培养基为Hib基础培养基,b培养基为发酵培养基,本实验为Hib疫苗培养基的优化和多糖抗原的纯化提供了实验依据.%Objective To analyze the protein and nucleic acid contents in fermentation supernatant of type b Haemophilus influenzae (Hib) using various media as well as the effect of additives in medium on fermentation product, and optimize the fermentation medium. Methods The media with various formula for shake-flask culture of Hib were designed, and the protein and nucleic acid contents in culture supernatant were determined, based on which an appropriate basal medium was selected for culture of Hib in fermenter, and the effects of various additives (such as phosphate, sodium salt, and metal ions) on the recovery of polysaccharide complex, morphology and color of fermentation supernatant of Hib were analyzed. Three

  11. 48株流感嗜血杆菌耐药性分析及β-内酰胺酶基因检测%Analysis on antimicrobial resistance and beta-lactamases gene detection of 48 haemophilus influenzae

    Institute of Scientific and Technical Information of China (English)

    桂和翠; 王中新; 沈继录

    2012-01-01

    目的 了解本地区流感嗜血杆菌的分布及耐药性,为指导临床合理用药提供依据.方法 k-B法进行药敏试验,玻片法测定β-内酰胺酶.PCR扩增TEM及ROB型β-内酰胺酶基因.结果 48株流感嗜血杆菌主要分布于呼吸内科和门诊.对复方新诺明、四环素和氨苄西林耐药率分别为62.50%、35.42% 和22 92%;阿莫西林/克拉维酸钾、氨曲南耐药率为12.50%;氨苄西林/舒巴坦、头孢噻肟、头孢拉定、头孢曲松、阿奇霉素、氯霉素耐药率为8.33%;头孢吡肟、头孢呋辛、环丙沙星、左氧氟沙星耐药率为6.25%.哌拉西林/三唑巴坦、亚胺培南敏感率高为100%.10株氨苄西林耐药菌株均产β-内酰胺酶,产酶率为20.83%,且均检测到TEM基因.结论 复方新诺明和四环素已不再适于临床治疗流感嗜血杆菌引起的感染.氨苄西林仍可作为临床经验用药.哌拉西林/三唑巴坦和亚胺培南抗菌活性高,可望作为治疗耐氨苄西林流感嗜血杆菌感染的理想用药.喹诺酮类药物耐药率高,应引起重视.流感嗜血杆菌对氨苄西林耐药的主要机制为产TEM型β-内酰胺酶.%To investigate antimicrobial resistance and genotypes of β-lactamase of in this erea , and guide clinical rational drug use effectively. Methods Kirby-Bauer method was applied for the drug susceptibility test and nitrocefin slide test was used to detect β-lactamase. The genotypes of β-lactamase were detected by PCR. Results A total of 48 strains of haemophilus influenzae were mainly distributed in department of respiratory and outpatient service. The resistant straint to compound sulf-amethoxazole, ampicillin and tetrocycline were 62. 50% , 35. 42% ,22. 92% respectively, the resistant rate to amoxicillin-clavulanic acid, ceftriaxome, aztreonam, ciprofloxa-cin and levofloxacin was 12.50% ,the resistant rate to ampicillin-sulbactam, cefotaxime, ceftazidime, cefuroxime, azithromycin, chlorampheniol was 8. 33% ;the

  12. Genetic diversity of the ftsI gene in β-lactamase-nonproducing ampicillin-resistant and β-lactamase-producing amoxicillin-/clavulanic acid-resistant nasopharyngeal Haemophilus influenzae strains isolated from children in South Korea.

    Science.gov (United States)

    Park, Chulmin; Kim, Kyung-Hyo; Shin, Na-Young; Byun, Ji-Hyun; Kwon, Eun-Young; Lee, Jae-Wook; Kwon, Hyo Jin; Choi, Eu Yoon; Lee, Dong-Gun; Sohn, Woo Yun; Kang, Jin Han

    2013-06-01

    Haemophilus influenzae frequently colonizes the nasopharynx of children and adults, which can lead to a variety of infections. We investigated H. influenzae carriage in the nasopharynx of 360 children, in terms of (1) the prevalence of strains with decreased susceptibility, and (2) the presence of amino acid substitutions in PBP3. One hundred twenty-three strains were isolated (34.2%, 123/360), 122 of which were classified as nontypable H. influenzae (NTHi). Of these, β-lactamase-nonproducing ampicillin-susceptible strains accounted for 26.2%, β-lactamase-producing-ampicillin-resistant strains for 9.0%, β-lactamase-nonproducing ampicillin-resistant (BLNAR) strains for 40.2%, and β-lactamase-producing amoxicillin-/clavulanic acid-resistant (BLPACR) for 24.6%, respectively. Pulsed field gel electrophoresis (PFGE) patterns were so diverse that they were clustered into 41 groups. The amino acid substitutions in the transpeptidase domain (292 amino acids) of ftsI in BLNAR isolates showed that group IIb accounted for 30.6%, IIc for 8.2%, IId for 16.3%, III for 32.7%, and the others for 12.2%. Moreover, groups IIb (56.7%; 17/30) and III (23.3%; 7/30) were prevalent among BLPACR strains. They were subclassified into more diverse sequence subtypes by analysis of the entire PBP3 (610 amino acids). Groups IIb, IIc, IId, and III exhibited 13, four, six, and four sequence subtypes, respectively. Such a genetic diversity is likely indicative of significant potential for decreased antimicrobial susceptibility in nasopharyngeal-colonizing NTHi strains.

  13. Etiology of bacterial meningitis among children aged 2-59 months in Salvador, Northeast Brazil, before and after routine use of Haemophilus influenzae type b vaccine Etiologia da meningite bacteriana em crianças com idade entre 2 e 59 meses em Salvador, Nordeste do Brasil, antes e depois do uso rotineiro da vacina para Haemophilus influenzae tipo b

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    Cristiana M. Nascimento-Carvalho

    2004-06-01

    Full Text Available OBJECTIVE: To describe the frequency of etiologic agents of bacterial meningitis (BM among children aged 2-59 months in a sample of patients in Salvador, Northeast Brazil, with emphasis on the frequency of BM of unknown etiology (BMUE, just before, during and after the implementation of routine immunization of infants with Haemophilus influenzae type b (Hib vaccination. METHOD: Demographic, clinical and cerebrospinal fluid (CSF information was collected from the chart of every patient, aged 2-59 months, whose CSF exam was performed at the CSF Lab - José Silveira Foundation, between January 1989 and December 2001. Every CSF exam was completely performed according to standard methods. The etiologic diagnosis was based on either culture and/or latex-agglutination test. When the agent was only seen on Gram stained smear, the diagnosis was descriptive. BMUE was defined as: glucose 100 mg / dl, white blood cell count > 20 cells / mm³, percentage of neutrophils > 80%. RESULTS: Of 1519 patients, 894 (58.9% had normal exams and BM was diagnosed in 95 (6.2%. Etiologic agents were: Hib (44.2%, meningococcus (13.7%, Gram-negative bacilli (11.6%, Mycobacterium tuberculosis (6.3%, pneumococcus (4.2%, other agents (4.2%; BMUE was diagnosed in 15.8% of cases with BM. By analysing the frequency of BMUE and Hib among all exams performed yearly, the peaks were recorded in 1989 (5.3% and 1990 (16.9%, respectively, decreasing to 0.7% and 0% in 2001. CONCLUSION: It is possible that the implementation of the conjugate Hib vaccine during the 1990's has been decreasing not only the occurrence of Hib meningitis but also of BMUE.OBJETIVO: Descrever a freqüência dos agentes etiológicos de meningite bacteriana (MB em amostra das crianças com idade entre 2 e 59 meses, em Salvador, Nordeste do Brasil, com ênfase na freqüência de MB de etiologia indeterminada (MBEI, antes, durante e após a implementação da imunização rotineira de lactentes com vacina para

  14. Competitive ELISA Method for Determining Haemophilus Influenza Type b Polysaccharide%b型流感嗜血杆菌多糖竞争ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    江山; 刘威; 兰芳; 廖红梅; 李小波; 张立平; 崔长法

    2013-01-01

    Objective To develop competitive ELISA method for determining the concentration of haemophilus influenza type b polysaccharide (PRP).Method PRP-Ty was set as coating antigen and poliasccharide as competition antigen,comparison were made between the two reactions against antisera of haemophilus influenza type b.The accuracy and precision of the method were validated accordingly.Results The optimal coating concentration of PRP-Ty and serum dilution factor was 1 ∶ 400,and 1 ∶ 40K respectively.Quantitation of PRP concentration was linear from 6.25ug/ml-100ug/ml,with lower limit of detection being 3.13ug/ml.The regression equation was B/Bo =-34.328 [PRP] + 105.03 with RSQ =0.995.The coefficient of variation of intra-assay and inter-assay was 3.4%-6.5% and 8.48% respectively.The recovery rate of PRP in culture medium was 95.7%.Conclusion The competitive ELISA method showed both high accuracy and precision,and thus can be used for detection of haemophilus influenza type b polysaccharide.%目的 建立竞争酶联免疫吸附分析法(Enzyme-Linked Immunosorbent Assay,ELISA),测定b型流感嗜血杆菌多糖(PRP)浓度方法以b型流感嗜血杆菌多糖-酪胺(Ty)为包被抗原,待测多糖为竞争抗原,与优化的抗b型流感嗜血杆菌多糖抗体反应,建立标准曲线,并验证该方法的准确性和精密度.结果 优化后的b型流感嗜血杆菌多糖-酪胺包被浓度为1∶400倍稀释,抗多糖血清稀释度为1∶40K.检测线性范围6.25μg/ml~100μg/ml,最低检测限为3.13μg/ml,回归方程为B/B0=-34.328[PRP]+105.03,线性相关系数为R2 =0.995.批内精密度为3.4%~6.5%,批间精密度为8.48%,测定培养基中b型流感嗜血杆菌多糖的回收率为95.7%.结论 本研究建立的b型流感嗜血杆菌多糖竞争ELISA方法准确性和精密性均较好,可以特异性地检测b型流感嗜血杆菌多糖浓度.

  15. Development of a Quantitative ELISA Method for Total Antibody of Haemophilus influenzae Type b%b型流感嗜血杆菌总抗体ELISA定量检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    李红; 李茂光; 唐静; 梁丽; 李亚南; 何莉; 叶强

    2011-01-01

    Objective To develop and preliminarily verify a quantitative ELISA method for total antibody of Haemophilus influenzae type b (Hib). Methods The coating concentration of antigen, working concentration of enzyme-labeled antibody, linear range of reference serum and substrate were optimized by chessboard titration, based on which the developed method was verified for specificity, accuracy and precision. Results The optimal coating concentration of antigen was 0. 4 μg/ml, and the optimal dilution of enzyme-labeled antibody was 1: 6 000. The optimal linear range of reference serum was 5 ~ 100 ng/ml (R2 = 0.997 6). TMB was selected as substrate. The developed method showed high specificity, accuracy and precision. Conclusion A quantitative ELISA method for total antibody of Hib was successfully developed, which might substitute to the determination of anti-Hib IgG in sera.%目的 建立定量检测血清中b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)总抗体的ELISA方法,并进行初步验证.方法 采用棋盘滴定法确定抗原最佳包被浓度及酶标抗体工作浓度,参考血清的线性范围及适合的底物,并验证方法的特异性、准确性及精密性.结果 最佳抗原包被浓度为0.4 μg/ml;最佳酶标抗体稀释度为1:6000;参考血清的最适线性范围为5~100ng/ml,RZ=0.9976 ;选择TMB作为底物.该方法特异性较好,准确性和精密性较高.结论 已成功建立了定量测定血清中Hib总抗体的ELISA方法,可逐渐替代测定血清中抗Hib IgG的方法.

  16. 用于Hib结合疫苗生产的新型候选菌株的评价%Evaluation on a new candidate strain for Haemophilus influenzae type b conjugate vaccine production

    Institute of Scientific and Technical Information of China (English)

    王伟; 马雷钧; 王月红; 朱为; 马相虎

    2011-01-01

    目的 观察b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)760705株在连续传代过程中的稳定性.方法 将Hib 760705株工作种子批菌种连续传代,对第5、第8、第10代Hib培养物进行全面检测,包括培养特性(细菌培养、卫星试验)、染色镜检、生化反应,以检测第5、第8、第10代Hib的生物学特性.同时采用血清凝集试验和聚合酶链反应荚膜分型方法进行b型荚膜多糖稳定性检测.结果 Hib 760705株工作种子批培养物在连续传代过程中具有典型的细菌学特性,能够稳定地产生b型荚膜多糖.结论 Hib 760705株有明确的来源和背景,可以稳定传代,具备作为Hib结合疫苗生产用候选菌株的条件.%Objective To observe the passage stability of Haemophilus influenzae type b(Hib) strain 760705. Methods Hib strain 760705 was cultured for 10 passages from the working seed lot, and the subcultures of the 5th, 8th and 10th passages were detected for biological charactristics comprehensively,including cultural characteristics( bacterial culture and satellite test), staining and microscopic examination,and biochemical reactions. Serological agglutination test and polymerase chain reaction for capsular typing were applied to confirm the generation stability of type b capsular polysaccharide. Results The subcultures of Hib strain 760705 had typical bacteriological characteristics and capability of yielding type b capsular polysaccharide. Conclusions Hib strain 760705 has a clear origin and background and can be subcultured stably, thus suggesting that it can be a candidate strain for Hib conjugate vaccine production.

  17. Eficácia do moxalactam no tratamento de meningites purulentas causadas por Haemophilus infuenzae e Neisseria meningitidis Efficacy of moxalactam in the treatment of purulent meningitis caused by Haemophilus influenzae and Neisseria meningitidis

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    Hagamenon R. da Silva

    1984-03-01

    Full Text Available Foi avaliada a eficácia do moxalactam no tratamento de meningites em crianças, causadas por H. influenzae (27 casos e N.meningitidis (6 casos. Dos 33 doentes tratados na dose de 100mg/Kg de peso (dose de ataque e 50mg de 12/12 horas por via venosa, 32 curaram-se. A tolerância ao produto foi muito boa, havendo alterações transitórias de transaminases e fosfatase alcalina; em um caso, houve hematoma posapendectomia, provavelmente relacionado ao uso deste antibiotico. Os níveis séricos e liquóricos do produto foram elevados; as concentrações no liquor excederam de muito a concentração bactericida mínima dos germes infectantes. O moxalactam se mostrou seguro e eficaz como terapia primária da meningite causada por H. influenzae e N.meningitidis em crianças.The clinical efficacy and safety of Moxalactam in purulent bacterial meningitis in children caused by H. influenzae (27 patients and N. meningitidis (6 patients was tested in a randon uncontrolled study. Clinical response was considered excelent, with cure of 32 of 33 patients. High levels of Moxalactam were achieved in the blood and cerebro-spinal fluid, with concentrations largely exceeding the minimum bacterial concentration (MIC for the infecting organisms. Tolerance was considered good, with only transient increases of transaminases and alkaline phosphatase in some patients; also, one patient developed a wound hematoma possibly related to Moxalactam therapy.

  18. CASE REPORT OF CEREBRAL ATROPHY INDUCED BY HAEMOPHILUS INFLUENZA TYPE B CONJUGATE VACCINE%b型流感嗜血杆菌结合疫苗致脑萎缩病例报告

    Institute of Scientific and Technical Information of China (English)

    王仁富; 廖红英

    2012-01-01

    [目的]对接种某种b型流感嗜血杆菌结合疫苗后发生脑萎缩的个案进行分析,结合该个案提出对避免预防接种异常反应工作提出建议.[方法]个案分析.[结果]患儿的脑萎缩是接种疫苗后发生抽搐、发热、意识障碍等症状产生的后遗症.[结论]预防接种工作人员在接种疫苗时须加强安全意识,避免异常反应的发生.%[Objective] To analyze the cases with brain atrophy induced by vaccination with Haemophilus influenzae type b vaccine, combined with the case put forward to avoid abnormal reaction to the work of vaccination recommendations. [Methods] Case report analysis was taken. [RssultS] Children with brain atrophy is the occurrence of seizures after vaccination, fever, disturbance of consciousness and olher symptoms resulting sequela. [Conclusion] Vaccination of staff should be vaccinated to enhance safety awareness and prevent the occurrence of abnormal reaction.

  19. Serum antibody response to three non-typeable Haemophilus influenzae outer membrane proteins during acute otitis media and nasopharyngeal colonization in otitis prone and non-otitis prone children.

    Science.gov (United States)

    Kaur, Ravinder; Casey, Janet R; Pichichero, Michael E

    2011-01-29

    Non-typeable Haemophilus influenzae (NTHi) is the most common bacteria responsible for episodic acute otitis media (AOM; non-otitis prone), recurrent AOM (rAOM; otitis prone) and AOM treatment failure (AOMTF) in children. In this 3.5 years of prospective study, we measured the serum antibody response to outer membrane proteins D, P6 and OMP26 of NTHi in children with AOM (n=26), rAOM (n=32), AOMTF (n=27). The geometric mean titers (GMTs) of IgG at their acute AOM visit against Protein D in otitis prone children were significantly lower compared to AOMTF (p value4 fold increases in the non-otitis prone children (p value<0.001). We conclude that otitis prone children mount less of an IgG serum antibody response toward Protein D, P6 and OMP26 after AOM which may account for recurrent infections. The data on acute sera of otitis prone vs non-otitis prone children and the acute-to-convalescence response in non-otitis prone children point to a possible link of anti-PD to protection. Moreover, the data suggest that otitis prone children should be evaluated for their responses to Protein D, P6 and OMP26 vaccine antigens of NTHi.

  20. Interacción del componente pertussis de células completas con los antígenos tetánico, Haemophilus influenzae tipo B y hepatitis B en ensayos de potencia para vacunas combinadas

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    Mario Landys

    2008-04-01

    Full Text Available Las vacunas combinadas permiten un grado de aceptación mayor entre la población, dado que se requiere de menos inmunizaciones para proteger contra las enfermedades infecciosas. Sin embargo, esto ha generado nuevos retos, ya que se han reportado numerosas interacciones entre los diferentes antígenos que conforman estas vacunas. El propósito del presente trabajo fue evaluar la interferencia potencial del componente pertussis de células completas sobre los ensayos para determinar la actividad biológica de otros antígenos como toxoide tetánico, Haemophilus influenzae tipo b (Hib y hepatitis B. Para ello se estudiaron mediante ensayos de potencia vacunas combinadas que contenían estos antígenos y se compararon con vacunas monovalentes. A su vez se prepararon adyuvaciones experimentales cuya composición permitió estimar adecuadamente la extensión y naturaleza de la interacción entre componentes. Se obtuvo que el componente pertussis incrementó significativamente la actividad biológica de Hib y tétanos, aunque esto puede depender mucho del modelo animal y el diseño experimental utilizado. En cuanto al antígeno de hepatitis B se demostró que pertussis inhibía la potencia de este antígeno, tanto in vitro como in vivo, aunque por mecanismos diferentes y de forma no significativa.

  1. Detection of Streptococcus pneumoniae and Haemophilus influenzae type B by real-time PCR from dried blood spot samples among children with pneumonia: a useful approach for developing countries.

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    Laura Selva

    Full Text Available Dried blood spot (DBS is a reliable blood collection method for storing samples at room temperature and easily transporting them. We have previously validated a Real-Time PCR for detection of Streptococcus pneumoniae in DBS. The objective of this study was to apply this methodology for the diagnosis of S. pneumoniae and Haemophilus influenzae b (Hib in DBS samples of children with pneumonia admitted to two hospitals in Mozambique and Morocco.Ply and wzg genes of S. pneumoniae and bexA gene of Hib, were used as targets of Real-Time PCR. 329 DBS samples of children hospitalized with clinical diagnosis of pneumonia were tested.Real-Time PCR in DBS allowed for a significant increase in microbiological diagnosis of S. pneumoniae and Hib. When performing blood bacterial culture, only ten isolates of S. pneumoniae and none of Hib were detected (3·0% positivity rate, IC95% 1·4-5·5%. Real-Time PCR from DBS samples increased the detection yield by 4x fold, as 30 S. pneumoniae and 11 Hib cases were detected (12·4% positivity rate, IC95% 9·0-16·5%; P<0·001.Real-Time PCR applied in DBS may be a valuable tool for improving diagnosis and surveillance of pneumonia caused by S. pneumoniae or Hib in developing countries.

  2. 不可分型流行性感冒嗜血杆菌致病机制研究进展%Progress Toward Molecular Determinants of the Pathogenesis of Disease Due to Nontypeable Haemophilus Influenzae

    Institute of Scientific and Technical Information of China (English)

    田国忠; 张砺; 邵祝军

    2009-01-01

    流行性感冒(流感)嗜血杆菌(Haemophilus Influenzae,Hi)分为有荚膜的可分型(a~f型)和无荚膜的不可分型菌株,有荚膜的b型Hi(Hi Type b,Hib)是引起侵袭性感染,包括脑膜炎、肺炎的主要病原体,但Hib的感染有疫苗可以预防.不可分型Hi是导致儿童感染的重要病原体之一,可引起多种疾病,包括肺炎、中耳炎等,目前无疫苗预防.现将国内外对不可分型Hi的致病因子和致病机理进行综述,为疫苗的研制、疾病的治疗和预防控制提供帮助.

  3. IMMUNOGENICITY AND SAFETY OF QUINVAXEM® (DIPHTHERIA, TETANUS, WHOLE-CELL PERTUSSIS, HEPATITIS B AND HAEMOPHILUS INFLUENZAE TYPE B VACCINE) GIVEN TO VIETNAMESE INFANTS AT 2 TO 4 MONTHS OF AGE.

    Science.gov (United States)

    Huu, Tran Ngoc; Phuong, Nguyen Thi Minh; Toan, Nguyen Trong; Thang, Ho Vinh

    2015-07-01

    Vietnam plans to replace the routine childhood diphtheria, pertussis and tetanus combination (DPT) vaccine with a pentavalent vaccine. The present study was performed to assess the immunogenicity and safety of the combined diphtheria, tetanus, whole-cell pertussis, hepatitis B (HepB), and Haemophilus influenzae type b (Hib) (DTwP-HepB-Hib) Quinvaxem® vaccine in children. A total of 131 infants received the Quinvaxem® vaccine at 2, 3 and 4 months. Antibody levels were measured at baseline, at one month after the third injection and one year after the first injection. Seroprotection rates were high for each vaccine antigen at one month after the third dose: 93.1% for diphtheria, 98.5% for tetanus, 99.2% for pertussis (seroconversion rate), 93.1% for HepB, and 100% for Hib (anti-PRP ≥ 0.15 µg/ml). The rate of children with protective antibodies persisting at one year after the first dose was 88.4% for diphtheria, 49.6% for pertussis, 82.2% for tetanus, 76.7% for HepB and 97.7% for Hib (anti-PRP ≥ 0.15 µg/ml). The Quinvaxem® vaccine was well tolerated and has a low rate of adverse events. Quinvaxem® given at 2, 3 and 4 months of age was immunogenic and safe for primary immunization among infants in Vietnam.

  4. Drug-resistance and serotype of 807 strains of Haemophilus influenzae%807株流感嗜血杆菌的血清分型及耐药性的研究

    Institute of Scientific and Technical Information of China (English)

    钟天鹰; 迟富丽; 王惠云; 徐飞; 谈华; 陈倩; 胡正

    2008-01-01

    目的 了解南京地区儿童感染流感嗜血杆菌(Haemophilus influenza, Hi)的血清型及抗生素的耐药性.方法 采集南京儿童医院2004年6月至2007年6月20 985份不同种类标本,分离鉴定Hi 807例,玻片凝集法进行血清分型,纸片扩散法(K-B法)进行抗茵药物敏感实验,E-test法测最低抑菌浓度(MIC).结果 807株流感嗜血杆菌,血清分型,不定型(NTHi)47.71%,可分型中f型最多,占可分型36.02%,b型最少,仅占可分型1.66%.K-B法检测抗生素的耐药率,氨苄西林耐药率49.07%.结论 南京地区儿童感染Hi,可分型株中主要血清型以f、a型占多数.氨苄西林的耐药率逐年上升.

  5. Accumulation of Regulatory T Cells and Chronic Inflammation in the Middle Ear in a Mouse Model of Chronic Otitis Media with Effusion Induced by Combined Eustachian Tube Blockage and Nontypeable Haemophilus influenzae Infection.

    Science.gov (United States)

    Hirano, Takashi; Kodama, Satoru; Kawano, Toshiaki; Suzuki, Masashi

    2016-01-01

    Nontypeable Haemophilus influenzae (NTHi) is associated with chronic otitis media (COM). In this study, we generated a murine model of COM by using eustachian tube (ET) obstruction and NTHi (10(7) CFU) inoculation into the tympanic bulla, and we investigated the relationship between regulatory T cells (Treg) and chronic inflammation in the middle ear. Middle ear effusions (MEEs) and middle ear mucosae (MEM) were collected at days 3 and 14 and at 1 and 2 months after inoculation. Untreated mice served as controls. MEEs were used for bacterial counts and to measure the concentrations of cytokines. MEM were collected for histological evaluation and flow cytometric analysis. Inflammation of the MEM was prolonged throughout this study, and the incidence of NTHi culture-positive MEE was 38% at 2 months after inoculation. The levels of interleukin-1β (IL-β), tumor necrosis factor alpha, IL-10, and transforming growth factor β were increased in the middle ear for up to 2 months after inoculation. CD4(+) CD25(+) FoxP3(+) Treg accumulated in the middle ear, and the percentage of Treg in the MEM increased for up to 2 months after inoculation. Treg depletion induced a 99.9% reduction of bacterial counts in MEEs and also significantly reduced the ratio of NTHi culture-positive MEE. The levels of these cytokines were also reduced in MEEs. In summary, we developed a murine model of COM, and our findings indicate that Treg confer infectious tolerance to NTHi in the middle ear.

  6. 2006-2011年浙江省杭州市萧山区流感嗜血杆菌耐药性监测%Surveillance of drug resistance of Haemophilus influenzae in Hangzhou.Zhejiang, 2006 -2011

    Institute of Scientific and Technical Information of China (English)

    郭远瑜; 吴伟志; 沈丽芳; 董华丽

    2012-01-01

    目的 调查了解流感嗜血杆菌的临床分布及耐药情况,为临床用药提供依据.方法 对2006年1月至2011年12月浙江萧山医院临床分离的流感嗜血杆菌,用ATB嗜血杆菌药敏板条测定抗菌药物的敏感性,用头孢硝噻吩纸片法测定β-内酰胺酶,所有数据用WHONET 5.6软件进行回顾性分析.结果 共检出流感嗜血杆菌375株,对复方新诺明、氨苄西林耐药分别为248株(66.1%)和127株(33.8%),对氯霉素、头孢克洛和四环素耐药分别为38株(10.2%)、52株(13.8%)和55株(14.7%);其他几种常用药物阿莫西林/克拉维酸、头孢呋辛、头孢噻肟、利福平和氧氟沙星对流感嗜血杆菌保持较好的抗菌活性(耐药率≤5.0%);β-内酰胺酶阳性菌株对多种药物的耐药率显著高于阴性菌株(P<0.01).结论 复方新诺明耐药率高而不宜用于流感嗜血杆菌感染的治疗,氨苄西林应慎重用于经验治疗;流感嗜血杆菌对氨苄西林耐药率呈下降趋势,对二代头孢菌素耐药率呈上升趋势,临床应根据药敏结果合理使用抗菌药物.%Objective To understand the clinical distribution and drug resistance of Haemophilus influenzae in Xiaoshan district in Hangzhou and provide evidence for clinical antibiotic use. Methods The drug susceptibility of H. influenzae strains isolated in Zhejiang Xisoahn hospital from January 2006 to December 2011 was tested by using ATB HAEMO strip, β-lactamases was detected with Nitrocefin discs test, and the results were analyzed retrospectively by using WHONTET 5. 6 software. Results A total of 375 H. influenzae isolates were tested, the resistance rate was 66.1% to trimethoprim-sulfamethoxazole (TMP/SMZ), 33.8% to ampicillin, 10.2% to chloramphenicol, 13.8% to cefaclor and 14.7% to tetracycline. Amoxicillin/clavulanic acid, cefuroxime, cefotaxime, rifampicin and ofloxac in had good antibacterial activity on H. influenzae (the resistance rate ≤5.0% ). The

  7. Influenza

    OpenAIRE

    Forleo-Neto Eduardo; Halker Elisa; Santos Verônica Jorge; Paiva Terezinha Maria; Toniolo-Neto João

    2003-01-01

    A influenza (gripe) é doença infecciosa aguda de origem viral que acomete o trato respiratório e a cada inverno atinge mais de 100 milhões de pessoas na Europa, Japão e Estados Unidos, causando anualmente a morte de cerca de 20 a 40 mil pessoas somente neste último país. O agente etiológico é o Myxovirus influenzae, ou vírus da gripe. Este subdivide-se nos tipos A, B e C, sendo que apenas os do tipo A e B apresentam relevância clínica em humanos. O vírus influenza apresenta altas taxas de mut...

  8. Effectiveness of Haemophilus influenzae Type b Conjugate Vaccine Introduction Into Routine Childhood Immunization in Kenya%B型流感嗜血杆菌结合疫苗纳入肯尼亚儿童计划免疫的效果

    Institute of Scientific and Technical Information of China (English)

    Karen D. Cowgill; Moses Ndiritu; Joyce Nyiro; 朱丽琳

    2007-01-01

    背景:迄今为止,非洲国家未将B型流感嗜血杆菌(Haemophilus influenzae Type b,Hib)结合疫苗列为首要的公共卫生问题,其原因在于有关Hib疾病负担以及Hib疫苗效力的资料仍很稀缺。2001年,肯尼亚的婴儿开始接种Hib疫苗。

  9. Classification of a Haemophilus influenzae ABC transporter HI1470/71 through its cognate molybdate periplasmic binding protein, MolA.

    Science.gov (United States)

    Tirado-Lee, Leidamarie; Lee, Allen; Rees, Douglas C; Pinkett, Heather W

    2011-11-01

    molA (HI1472) from H. influenzae encodes a periplasmic binding protein (PBP) that delivers substrate to the ABC transporter MolB(2)C(2) (formerly HI1470/71). The structures of MolA with molybdate and tungstate in the binding pocket were solved to 1.6 and 1.7 Å resolution, respectively. The MolA-binding protein binds molybdate and tungstate, but not other oxyanions such as sulfate and phosphate, making it the first class III molybdate-binding protein structurally solved. The ∼100 μM binding affinity for tungstate and molybdate is significantly lower than observed for the class II ModA molybdate-binding proteins that have nanomolar to low micromolar affinity for molybdate. The presence of two molybdate loci in H. influenzae suggests multiple transport systems for one substrate, with molABC constituting a low-affinity molybdate locus. PMID:22078568

  10. Research Progress in Protective Antigens of Nontypeable Haemophilus Influenzae%不定型流感嗜血杆菌保护性抗原研究进展

    Institute of Scientific and Technical Information of China (English)

    冯旭敏; 华春珍

    2011-01-01

    随着b型流感嗜血杆菌(Haemphilus influenzae tybe b,Hib)菌苗在世界范围的广泛接种,全球各地的Hib感染率已大幅度下降.而不定型流感嗜血杆菌(Nontypeable-Haemphilus influenzae,NTHi)的比例大幅上升,已成为主要的Hi致病株,常引起中耳炎、急性上呼吸道感染、肺炎、败血症等疾病.针对NTHi的预防,目前尚无NTHi疫苗产品问世.文章就NTHi疫苗的研究现状、主要保护性疫苗的实验研究进展作一综述.

  11. Classification of a Haemophilus influenzae ABC Transporter HI1470/71 through Its Cognate Molybdate Periplasmic Binding Protein, MolA

    Energy Technology Data Exchange (ETDEWEB)

    Tirado-Lee, Leidamarie; Lee, Allen; Rees, Douglas C.; Pinkett, Heather W. (CIT); (NWU)

    2014-10-02

    molA (HI1472) from H. influenzae encodes a periplasmic binding protein (PBP) that delivers substrate to the ABC transporter MolB{sub 2}C{sub 2} (formerly HI1470/71). The structures of MolA with molybdate and tungstate in the binding pocket were solved to 1.6 and 1.7 {angstrom} resolution, respectively. The MolA-binding protein binds molybdate and tungstate, but not other oxyanions such as sulfate and phosphate, making it the first class III molybdate-binding protein structurally solved. The {approx}100 {mu}M binding affinity for tungstate and molybdate is significantly lower than observed for the class II ModA molybdate-binding proteins that have nanomolar to low micromolar affinity for molybdate. The presence of two molybdate loci in H. influenzae suggests multiple transport systems for one substrate, with molABC constituting a low-affinity molybdate locus.

  12. b型流感嗜血杆菌多糖竞争ELISA检测方法的建立与应用%Establishment and application of competitive ELISA method for detertion of Haemophilus influenzae type b polysaccharide

    Institute of Scientific and Technical Information of China (English)

    刘威; 江山; 杨溢尧; 李小波; 兰芳; 崔长法

    2014-01-01

    目的 建立检测b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)多聚核糖基核糖醇磷酸盐(polyribosylribitol phosphate,PRP)的竞争酶联免疫吸附法(competative enzyme-linked immunosorbent assay,cELISA).方法 以Hib PRP-酪胺(PRP-Ty)为包被抗原,待测PRP为竞争抗原,利用方阵滴定法确定抗原与血清抗PRP抗体的最适反应条件,建立cELISA法并验证其准确性和精密度,并将该法与传统检测法进行比较.结果 最适包被PRP-Ty浓度为1.30 mg/L,血清抗PRP抗体稀释度为1∶40 000.该法的检测线性范围为6.25~100.00 mg/L,决定系数(R2)为0.99,批内精密度为3.39%~6.53%,批间精密度为8.48%.该法对Hib PRP的检测结果与传统化学法一致.结论 建立的Hib多糖cELISA法的准确性和精密性良好,可特异性检测Hib PRP.%Objective To establish competitive ELISA (cELISA) method for determining polyribosylribitol phosphate (PRP) of Haemophilus influenzae type b (Hib).Methods PRP-Ty was used as coating antigen and Hib PRP sample was used as competitive antigen.Optimal reaction conditions were determined between antigens and serum antibodies to Hib PRP by criss-cross serial dilution analysis.The cELISA method was established and its accuracy and precision were validated.The cELISA method was compared with the traditional method.Results Optimal coating concentration of PRP-Ty and antiserum dilution were 1.30 mg/L and 1 ∶ 40 000,respectively.The linear range of the cELISA method was from 6.25 mg/L to 100.00 mg/L,and the coefficient of determination (R2) was 0.99.The precision of intraassay and inter-assay were 3.39%-6.53% and 8.48%,respectively.The detection results of Hib PRP with the cELISA method were consistent with those with the traditional chemical method.Conclusion The cELISA method has better accuracy and precision,and can be used for detection of Hib polysaccharide.

  13. 十堰市健康人群流感嗜血杆菌携带状况分析%Analysis of the prevalence of carriers of haemophilus influenza among the health population in the City of Shiyan

    Institute of Scientific and Technical Information of China (English)

    杨红梅; 吕静; 李国明; 高景枝; 郑向梅; 王滨

    2012-01-01

    Objectives To ascertain the prevalence of carriers of Haemophilus influenza (Hi) in the mouth and throat a-mong the healthy population in the city of Shiyan and to provide evidence for effective prevention and control of Hi infection. Methods Throat swab specimens were randomly collected from 202 healthy individuals from five age groups in the city of Shiyan. The prevalence of carriers of Hi was monitored using TaqMan quantitative PCR. Results Of the 202 specimens, 110 tested positive at a rate of 54. 46%. The specimens that tested positive most often (76. 92% , 50/65 specimens)were from children ages 3 and up. These children accounted for 45. 45% (50/110)of all positive specimens. The specimens that tested positive least often (15. 62%) were from young adults age 20 and over 20 years. There are the significant differences (X2 =40. 03, P<0. 01) in the positive rate for different age groups. Of the throat swab specimens that tested positive according to PCR, 52. 78% (57/108) were from males while 56. 38% (53/94) were from females. Positivity for Haemophilus influenza type B (Hib) was 8. 42% while positivity for other types and unknown types of Hi was 46. 04%. Conclusion The healthy population in the city of Shiyan tested positive for Hi at a relatively high rate. Hib vaccines should be emphasized and research on new vaccines should be enhanced.%目的 了解十堰市健康人群口咽部流感嗜血杆菌(Hi)携带情况,为有效防控Hi感染提供依据. 方法 随机抽取十堰市5个年龄组202份健康人群的咽拭子标本,Hi携带检测采用荧光定量PCR法. 结果 202份咽拭标本Hi阳性110份,阳性率为54.46%.其中3岁~年龄组阳性率最高,为76.92%(50/65),占总阳性数的45.45%(50/110);≥20岁人群阳性率最低,为15.62%;各年龄组人群Hi阳性率差异有统计学意义(x2=40.03,P<0.01);男、女咽拭子标本PCR阳性率分别为52.78%(57/108)和56.38%(53/94);b型流感嗜血杆菌(Hib)阳性率为8.42

  14. 冻干b型流感嗜血杆菌结合疫苗稳定性研究%Study on stability of a freeze-dried Haemophilus influenzae type b conjugate vaccine

    Institute of Scientific and Technical Information of China (English)

    袁军; 李新国

    2011-01-01

    目的 对以乳糖作为稳定剂的b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)结合疫苗冻干剂型进行稳定性研究.方法 选取3批冻干Hib结合疫苗,分别于2~8℃保存42个月,20~25℃保存7个月,37℃保存5周.并于考察期内对疫苗进行外观检查、检测回收率(KD <0.2)、游离多糖含量、水分和小鼠效力试验,观察其是否发生降解.结果 在考察期内,冻干疫苗小鼠效力试验阳转率均为100%,外观检查均符合规定,回收率(KD<0.2)均≥68%,游离多糖均≤18%,水分均≤3.0%.各项指标均达到中国药典要求.结论 冻干疫苗于2~8℃保存42个月,20~25℃保存7个月,37℃保存5周质量稳定.%Objective To research the stability of a freeze-dried Haemophilus influenzae type b (Hib)conjugate vaccine using lactose as a stabilizer.Methods Three batches of the freeze-dried Hib conjugate vaccine were selected to be stored at 2-8 ℃ for 42 months,20-25 ℃ for 7 months and 37 ℃ for 5 weeks,respectively.Tests for appearance,recovery rate of polysaccharide (KD < 0.2),free polysaccharide content,moisture content and mouse potency test were performed during observation to see whether degradation of the vaccine occurred.Results The seroconversion rate in mouse potency test was 100%,the appearance of vaccine was fit for the standard,recovery rate of polysaccharide (KD <0.2) was≥68%,free polysaccharide content≤ 18%,moisture content ≤3.0% for all three batches of the freeze-dried vaccine during observation.All the indexes of freeze-dried vaccine reach the requirements of Chinese Pharmacopeia.Conclusion The freeze-dried Hib conjugate vaccine has stable quality when stored at 2-8℃ for 42 months,20-25℃ for 7 months and 37℃ for 5 weeks.

  15. 4株荚膜型流感嗜血杆菌分型情况的比较%Comparison of typing among 4 haemophilus influenzae isolates with capsule

    Institute of Scientific and Technical Information of China (English)

    王复甦; 占利; 杨婷婷; 姚苹苹; 徐宝祥; 叶菊莲

    2012-01-01

    Objective:To provide fundamental information about bacterial typing for Hi - associated diseases control. Methods:Serotypes, biotypes, PFGE types and MLST(Multilocus Sequence Typing) types of 4 Haemophilus influenzae capsule strains isolated from children under 5 years old were analyzed. Results: It was found that the 2 Hib isolates were biotype Ⅰ, the Hie isolate was biotype Ⅲ , and the Hia/f which was cross reacted with antisera a and f capsule types belonged to biotype Ⅰ . The similarity of PFGE pattern among the 4 capsule Hi isolates varied between 48. 8 % ~ 58. 5 % , and the two biotype I Hibs only 48. 8 % . The ST types of 2 Hibs with the same biotype Ⅰ were both ST95 ,while the biotype Ⅲ Hie was ST - 18. The biotype Ⅰ Hia/f maybe a new ST type, waiting for submitting and verification. Conclusion: The association among 4 capsuled Hi is comparatively weak, suggesting thare is no dominant capsuled Hi type among human group. PFGE can be used to identify the source of Hi strains with the same serotype and biotype.%目的:为预防和控制Hi相关疾病提供菌株分型资料.方法:分析比较4株从5岁以下幼儿中分离的荚膜型流感嗜血杆菌(Haemophilus influenzae,Hi)的血清分型、生物分型、脉冲场凝胶电泳(PFGE)和多位点序列分型(MLST)的结果.结果:4株荚膜型Hi中,2株Hib均为生物Ⅰ型;1株Hie为生物Ⅲ型;1株与a、f荚膜型抗血清有交叉凝集的Hia/f为生物Ⅰ型.4株PFGE型的相似度,为48.8% ~ 58.5%,其中,2株Hib菌株之间的相似度仅为48.8%;MLST分型,2株Ⅰ型Hib同为ST-95型,1株生物Ⅲ型的Hie为ST-18型,1株生物Ⅰ型的Hia/f可能为新的ST型,等待提交验证.结论:4株荚膜型Hi彼此间遗传关联度小,提示人群中尚未形成优势荚膜型Hi的聚集.Hi当菌株生物型和血清型相同时,PFGE能较好地区分不同来源的菌株.

  16. How did international agencies perceive the avian influenza problem? The adoption and manufacture of the 'One World, One Health' framework.

    Science.gov (United States)

    Chien, Yu-Ju

    2013-02-01

    Over the past few years, a 'One World, One Health' (OWOH) policy framework has become the guiding principle for international responses to avian influenza and other zoonotic infectious diseases. Several specialised inter-governmental agencies, including the World Health Organization, the Food and Agricultural Organization and World Organization for Animal Health, jointly endorsed and promoted this framework. This article attempts to explain why international agencies advocated OWOH despite its vagueness. By examining how these international agencies gradually reframed avian flu problems, this article illustrates the crucial roles of international agencies in constructing disease knowledge and relevant policy responses. It shows that the three agencies adopted an all-inclusive approach to reduce conflicts, defend their legitimacy, and facilitate commitment for collaboration. Not only has this new framework reduced tensions between agencies, it has also reshaped the interests and interactions of other global actors. The case thus illustrates how these organisational actors actively manufacture new cognitive frames and policy regimes that advance their own legitimacy and influence. PMID:23095003

  17. Immunization coverage and risk factors for failure to immunize within the Expanded Programme on Immunization in Kenya after introduction of new Haemophilus influenzae type b and hepatitis b virus antigens

    Directory of Open Access Journals (Sweden)

    Feikin Daniel R

    2006-05-01

    Full Text Available Abstract Background Kenya introduced a pentavalent vaccine including the DTP, Haemophilus influenzae type b and hepatitis b virus antigens in Nov 2001 and strengthened immunization services. We estimated immunization coverage before and after introduction, timeliness of vaccination and risk factors for failure to immunize in Kilifi district, Kenya. Methods In Nov 2002 we performed WHO cluster-sample surveys of >200 children scheduled for vaccination before or after introduction of pentavalent vaccine. In Mar 2004 we conducted a simple random sample (SRS survey of 204 children aged 9–23 months. Coverage was estimated by inverse Kaplan-Meier survival analysis of vaccine-card and mothers' recall data and corroborated by reviewing administrative records from national and provincial vaccine stores. The contribution to timely immunization of distance from clinic, seasonal rainfall, mother's age, and family size was estimated by a proportional hazards model. Results Immunization coverage for three DTP and pentavalent doses was 100% before and 91% after pentavalent vaccine introduction, respectively. By SRS survey, coverage was 88% for three pentavalent doses. The median age at first, second and third vaccine dose was 8, 13 and 18 weeks. Vials dispatched to Kilifi District during 2001–2003 would provide three immunizations for 92% of the birth cohort. Immunization rate ratios were reduced with every kilometre of distance from home to vaccine clinic (HR 0.95, CI 0.91–1.00, rainy seasons (HR 0.73, 95% CI 0.61–0.89 and family size, increasing progressively up to 4 children (HR 0.55, 95% CI 0.41–0.73. Conclusion Vaccine coverage was high before and after introduction of pentavalent vaccine, but most doses were given late. Coverage is limited by seasonal factors and family size.

  18. Bacterial lysate of Haemophilus influenzae type b bacteria culture and crushing process parameter optimization%b型流感嗜血杆菌培养及细菌溶解产物制备工艺研究

    Institute of Scientific and Technical Information of China (English)

    徐道俊; 马波

    2014-01-01

    目的 研究b型流感嗜血杆菌(Haemophilus influenza type b,Hib)培养条件及细菌溶解产物制备工艺.方法 比较Hib在传统培养基和改良培养基的生长情况,优化改良培养基在生物反应器中对Hib培养条件(pH值、温度、溶解氧等),研究最佳菌体细胞破碎方法及细菌溶解产物纯化制备工艺.结果 改良培养基可代替传统培养基用于Hib的生产培养,且Hib在pH7.4、温度为36℃、溶解氧为25%的改良培养基中,生长迅速,菌体产量最高.高压匀浆破碎法的破碎效果明显优于超声波破碎法,破碎效果达到98%以上,纯化后的细菌溶解产物的多糖含量、总氮含量、蛋白质含量、核酸含量及细菌内毒素含量均符合《中华人民共和国药典》(三部)2010版中b型流感嗜血杆菌结合疫苗的质量标准.结论 初步建立了b型流感嗜血杆菌培养及细菌溶解产物制备工艺.

  19. Antimicrobial resistance analysis in Haemophilus influenzae strains isolated from children with respiratory tract infection%儿童呼吸道感染流感嗜血杆菌的耐药性分析

    Institute of Scientific and Technical Information of China (English)

    肖航; 吴祥林; 姚伟; 卢小娟; 陈俊玲; 王光磊; 李如凯

    2014-01-01

    目的 了解儿童下呼吸道感染中流感嗜血杆菌的分离率及耐药性,为临床治疗提供依据.方法 选择临床分离的流感嗜血杆菌173株,对其进行生物分型、血清分型及β-内酰胺酶测定,同时用K-B法做药敏试验.结果 流感嗜血杆菌阳性分离率为13.9%,生物Ⅱ型占32.0%、Ⅲ型占42.0%,血清b型为21.3%,β-内酰胺酶阳性率为34.4%.结论 流感嗜血杆菌是儿童下呼吸道感染的重要病原菌,其β-内酰胺酶阳性率较高,应引起临床重视.%Objective To understand the antimicrobial resistance and isolation rates of Haemophilus influenzae (Hi) from children with acute lower respiratory tract infection.Methods Hi was isolated and identified by biochemistry tests and serotyped by Hi antiserum.β-1actamase was tested by Cephalosporins tanner thiaphenol method and antimicrobial sensitivity was determined by Kirby-Bauer method.Results 173 strains were isolated from 1 248 sputum specimens,of which biological type Ⅱ,Ⅲ and serotype b were accounted for 32.0%,42.0% and 21.3%,respectively; the prevalence of β-lactamase was 34.4%.Conclusion Hi is the common pathogen of lower respiratory tract infection in children and higher rates of β-lactamase should be taken into account.

  20. 2011年至2012年某地区儿童呼吸道感染流感嗜血杆菌的季节分布及耐药性分析%Analysis of the Distribution and Drug Resistance of Haemophilus Influenza Season in Children's Respiratory Tract Infection in A Area 2011-2012

    Institute of Scientific and Technical Information of China (English)

    孙志豪; 袁满海; 李仕杰

    2013-01-01

      目的了解东莞地区儿童呼吸道感染流感嗜血杆菌的季节分布及其耐药性情况。方法对东莞市妇幼保健院2011年1月至2012年12月5522例门诊和住院呼吸道感染患儿痰液标本,进行流感嗜血杆菌培养,分离、鉴定和药敏,利用 WHONET5.5软件进行分析。结果流感嗜血杆菌的阳性标本数为532株(9.63%),β-内酰胺酶产酶率为37.22%;春季(37.2%)为儿童感染高峰期,夏季(29.9%)次之,秋(13.2%)冬(19.7%)最少;氧氟沙星、头孢噻肟、利福平的敏感率达到98.1%、98.7%、98.9%,而复方新诺明的耐药率为75.2%,氨苄西林的耐药率为45.1%。结论东莞地区儿童呼吸道感染流感嗜血杆菌较为多见,与季节有关,其β-内酰胺酶产酶率较高,多数抗生素对流感嗜血杆菌的敏感性高,而氨苄西林、复方新诺明的耐药率高,不适于流感嗜血杆菌的治疗。%  Objective To investigate the distribution and drug resistance situation of season of Haemophilus influenzae in children with respiratory tract infections in Dongguan area. Methods The maternal and child health hospital in Dongguan city in 2011 January to 2012 December 5522 cases of outpatients and inpatients sputum specimens of respiratory tract infection, Haemophilus influenzae culture, isolation, identification and drug sensitivity, were analyzed using WHONET5.5 software. Results The number of positive specimens of Hemophilus influenza for 532 strains (9.63%), β-lactamase enzyme production rate was 37.22%; the spring (37.2%) at the peak of infection for children, summer (29.9%) times, autumn and winter (13.2%), (19.7%) at least; the sensitive rate of cefotaxime, ofloxacin, Rifuku Hira reached 98.1%, 98.7%, 98.9%, and compound sulfamethoxazole resistance rates were 75.2%, ampicillin resistant rate was 45.1%. Conclusion More common Haemophilus influenzae in children with respiratory tract infection in Dongguan area

  1. Antibody Persistence in Young Children 5 Years after Vaccination with a Combined Haemophilus influenzae Type b-Neisseria meningitidis Serogroup C Conjugate Vaccine Coadministered with Diphtheria-Tetanus-Acellular Pertussis-Based and Pneumococcal Conjugate Vaccines

    Science.gov (United States)

    Tejedor, Juan Carlos; Brzostek, Jerzy; Konior, Ryszard; Grunert, Detlef; Kolhe, Devayani; Baine, Yaela

    2016-01-01

    We evaluated antibody persistence in children up to 5 years after administration of a combined Haemophilus influenzae type b (Hib)-Neisseria meningitidis serogroup C (MenC)-tetanus toxoid (TT) conjugate vaccine coadministered with a pneumococcal conjugate vaccine. This is the follow-up study of a randomized trial (ClinicalTrials.gov registration no. NCT00334334/00463437) in which healthy children were vaccinated (primary vaccinations at 2, 4, and 6 months of age and booster vaccination at 11 to 18 months of age) with Hib-MenC-TT or a control MenC conjugate vaccine, coadministered with diphtheria-tetanus-acellular pertussis (DTPa)-based combination vaccines (DTPa/Hib for control groups) and a pneumococcal conjugate vaccine (10-valent pneumococcal nontypeable H. influenzae protein D conjugate vaccine [PHiD-CV] or 7-valent cross-reacting material 197 [CRM197] conjugate vaccine [7vCRM]). MenC antibody titers were measured with a serum bactericidal antibody (SBA) assay using rabbit complement (i.e., rabbit SBA [rSBA]), and antibodies against Hib polyribosylribitol phosphate (PRP) were measured with an enzyme-linked immunosorbent assay. Antibody persistence up to 5 years after booster vaccination is reported for 530 children ∼6 years of age. The percentages of children with seroprotective rSBA-MenC titers were between 24.2% and 40.1% in all groups approximately 5 years after booster vaccination. More than 98.5% of children in each group retained seroprotective anti-PRP concentrations. No vaccine-related serious adverse events and no events related to a lack of vaccine efficacy were reported. Approximately 5 years after booster vaccination, the majority of children retained seroprotective anti-PRP antibody concentrations. The percentage of children retaining seroprotective rSBA-MenC titers was low (≤40%), suggesting that a significant proportion of children may be unprotected against MenC disease. (This study has been registered at ClinicalTrials.gov under

  2. Antibody Persistence in Young Children 5 Years after Vaccination with a Combined Haemophilus influenzae Type b-Neisseria meningitidis Serogroup C Conjugate Vaccine Coadministered with Diphtheria-Tetanus-Acellular Pertussis-Based and Pneumococcal Conjugate Vaccines.

    Science.gov (United States)

    Tejedor, Juan Carlos; Brzostek, Jerzy; Konior, Ryszard; Grunert, Detlef; Kolhe, Devayani; Baine, Yaela; Van Der Wielen, Marie

    2016-07-01

    We evaluated antibody persistence in children up to 5 years after administration of a combined Haemophilus influenzae type b (Hib)-Neisseria meningitidis serogroup C (MenC)-tetanus toxoid (TT) conjugate vaccine coadministered with a pneumococcal conjugate vaccine. This is the follow-up study of a randomized trial (ClinicalTrials.gov registration no. NCT00334334/00463437) in which healthy children were vaccinated (primary vaccinations at 2, 4, and 6 months of age and booster vaccination at 11 to 18 months of age) with Hib-MenC-TT or a control MenC conjugate vaccine, coadministered with diphtheria-tetanus-acellular pertussis (DTPa)-based combination vaccines (DTPa/Hib for control groups) and a pneumococcal conjugate vaccine (10-valent pneumococcal nontypeable H. influenzae protein D conjugate vaccine [PHiD-CV] or 7-valent cross-reacting material 197 [CRM197] conjugate vaccine [7vCRM]). MenC antibody titers were measured with a serum bactericidal antibody (SBA) assay using rabbit complement (i.e., rabbit SBA [rSBA]), and antibodies against Hib polyribosylribitol phosphate (PRP) were measured with an enzyme-linked immunosorbent assay. Antibody persistence up to 5 years after booster vaccination is reported for 530 children ∼6 years of age. The percentages of children with seroprotective rSBA-MenC titers were between 24.2% and 40.1% in all groups approximately 5 years after booster vaccination. More than 98.5% of children in each group retained seroprotective anti-PRP concentrations. No vaccine-related serious adverse events and no events related to a lack of vaccine efficacy were reported. Approximately 5 years after booster vaccination, the majority of children retained seroprotective anti-PRP antibody concentrations. The percentage of children retaining seroprotective rSBA-MenC titers was low (≤40%), suggesting that a significant proportion of children may be unprotected against MenC disease. (This study has been registered at ClinicalTrials.gov under

  3. A multiple touchdown PCR for detection of Streptococcus pneumonia,Haemophilus influenzae type b in sputum samples%多重降落PCR检测痰标本肺炎链球菌与b型流感嗜血杆菌

    Institute of Scientific and Technical Information of China (English)

    罗欲承; 杜蓬; 赵建忠; 段秀杰; 侯艳娇; 潘红; 邵世和

    2012-01-01

    目的:建立可同时检测痰标本中肺炎链球菌(Streptococcus pneumoniae,Sp)与b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)的多重降落PCR.方法:以Sp cpsA基因、Hib capⅡ区为靶标设计引物,并用PrimerBLAST在线软件检测其特异性,多重降落PCR检测492份痰标本,并以标准菌株作为对照,结果与细菌培养法进行比较.结果:所建立的多重降落PCR对Sp、Hib的检测下限分别达6.3pg、0.2 pg;与细菌培养法相比,灵敏度及总灵敏度均达100%,检测Sp、Hib特异性分别为94%、98%,总特异性达92%.结论:所建立的多重降落PCR具有高灵敏度与特异性,可作为Sp、Hib感染的快速诊断与流行病学研究的备选手段.%Objective: To develop a multiple touchdown PCR for simultaneous detection of Streptococcus pneumonia, Haemophilia influenzae type b in sputum samples. Methods: Primers based on cpsA, the region Ⅱ of cap were designed for Sp, Hib detection respectively, and their specificities were validated using Primer-BLAST. These primers were mixed in a multiple touchdown PCR amplification system and 492 sputum samples were detected. Standard strains were as controls and the results were compared with which of bacteria culture method. Results; The Sp and Hib detection limits of the multiple touchdown PCR assay were 6.3 pg, 0.2 pg respectively. The sensitivities of both genes were all 100% in contrast with bacteria culture method. The specificities of both genes were 94% , 98% respectively and total specificity was 92%. Conclusion: A higher sensitivity and specificity multiple touchdown PCR assay was developed, and it could be an alternative method for rapid clinical diagnosis and epidemiological study of Sp, and Hib infection.

  4. Antibody Persistence in Young Children 5 Years after Vaccination with a Combined Haemophilus influenzae Type b-Neisseria meningitidis Serogroup C Conjugate Vaccine Coadministered with Diphtheria-Tetanus-Acellular Pertussis-Based and Pneumococcal Conjugate Vaccines.

    Science.gov (United States)

    Tejedor, Juan Carlos; Brzostek, Jerzy; Konior, Ryszard; Grunert, Detlef; Kolhe, Devayani; Baine, Yaela; Van Der Wielen, Marie

    2016-07-01

    We evaluated antibody persistence in children up to 5 years after administration of a combined Haemophilus influenzae type b (Hib)-Neisseria meningitidis serogroup C (MenC)-tetanus toxoid (TT) conjugate vaccine coadministered with a pneumococcal conjugate vaccine. This is the follow-up study of a randomized trial (ClinicalTrials.gov registration no. NCT00334334/00463437) in which healthy children were vaccinated (primary vaccinations at 2, 4, and 6 months of age and booster vaccination at 11 to 18 months of age) with Hib-MenC-TT or a control MenC conjugate vaccine, coadministered with diphtheria-tetanus-acellular pertussis (DTPa)-based combination vaccines (DTPa/Hib for control groups) and a pneumococcal conjugate vaccine (10-valent pneumococcal nontypeable H. influenzae protein D conjugate vaccine [PHiD-CV] or 7-valent cross-reacting material 197 [CRM197] conjugate vaccine [7vCRM]). MenC antibody titers were measured with a serum bactericidal antibody (SBA) assay using rabbit complement (i.e., rabbit SBA [rSBA]), and antibodies against Hib polyribosylribitol phosphate (PRP) were measured with an enzyme-linked immunosorbent assay. Antibody persistence up to 5 years after booster vaccination is reported for 530 children ∼6 years of age. The percentages of children with seroprotective rSBA-MenC titers were between 24.2% and 40.1% in all groups approximately 5 years after booster vaccination. More than 98.5% of children in each group retained seroprotective anti-PRP concentrations. No vaccine-related serious adverse events and no events related to a lack of vaccine efficacy were reported. Approximately 5 years after booster vaccination, the majority of children retained seroprotective anti-PRP antibody concentrations. The percentage of children retaining seroprotective rSBA-MenC titers was low (≤40%), suggesting that a significant proportion of children may be unprotected against MenC disease. (This study has been registered at ClinicalTrials.gov under

  5. Classification of a Haemophilus influenzae ABC transporter HI1470/71 through its cognate molybdate periplasmic binding protein, MolA

    OpenAIRE

    Tirado-Lee, Leidamarie; Lee, Allen; Rees, Douglas C.; Pinkett, Heather W.

    2011-01-01

    molA(HI1472) from H. influenzae encodes a periplasmic binding protein (PBP) that delivers substrate to the ABC transporter MolB2C2 (formerly HI1470/71). The structures of MolA with molybdate and tungstate in the binding pocket were solved to 1.6 and 1.7-Å resolution, respectively. The MolA binding protein binds molybdate and tungstate but not other oxyanions such as sulfate and phosphate, making it the first class III molybdate binding protein structurally solved. The ~100 μM binding affinity...

  6. Analysis of Immune Responses and Serological Cross Reactivities among Vibrio cholerae O1,Shigella flexneri 2a and Haemophilus influenzae b

    Institute of Scientific and Technical Information of China (English)

    Fazle Rabbi; Chowdhury R.Ahsan; Nasreen Sultana; Tasmina Rahman; H.M.Al-Emran; M.Nizam Uddin; Mahbub Hossain; Kazi Selim Anwar; Mahmuda Yasmin; Jamalun Nessa

    2008-01-01

    Antigenic determinants expressed on the bacterial cell surface are of importance in the serological characterization and microbiological diagnosis.The bacterial strains carrying these identical or similar antigenic epitopes might react with antibodies produced against other strains.In this study,strong immunogenicity and antigenic cross reactivity were demonstrated among V.choleae O1,S.flexnerii 2a and H.influenzae b surface components.The enzyme linked immunosorbent assay (ELISA) results were supported by Western blot analysis,where at least 20 antigenic bands,were obtained in each of the reactions,when the surface components were reacted with the homologous antisera.The indirect ELISA results also demonstrated high degree of antigenic relatedness between the surface components of these species,where each surface component was reacted with the heterologous antisera. Western blot analysis also revealed cross reactions between the surface components suggesting common distribution of antigensepitopes in these bacteriaI species.This study,thus,gave a clear idea of the level of antigenic sharing and variations among the pathogenic V.cholerae O1,S.flexneri 2a and H.influenzae b strains, which in future,may help in selecting a proper candidate for vaccines and immunodiagnostics development.

  7. 多重PCR技术检测b型流感嗜血杆菌荚膜基因%Detection of type b capsule gene of Haemophilus influenzae by multiplex PCR

    Institute of Scientific and Technical Information of China (English)

    冯旭敏; 华春珍; 王超前; 项华冰; 来志超; 夏卫良; 王玲玲

    2012-01-01

    Objective To detect Haemophilus influen2ae type b strains isolated from children in Hangzhou by multiplex PCR methods. Methods All Haemophilus influenzae strains were analyzed by multiplex PCR. BexA-primers and b capsular type-specific gene primers were used as golden standard- Results Of all the 399 clinical strains isolated from children during the period from 2001-2002 and 2006-2007, 297 strains (74. 44%) were nontypeable, while 102 strains (25.56%) were typeable and only 1 (0.25%) of them was type b, respectively. PCR results showed that of all the 102 strains which showed positive with the slide agglutination method. 101 were positive and only 1 -was negative while 2 strains showed positive from the 297 strains which were nontypesble- The results showed consistent with the serotype with the sensitivity of 99.02% and specificity of 99.33%. Conclusions With genotyping success rate and detection rate for Hib reaching 100%, the investigation overcame the shortcomings of serotyping by the technology of bex-PCR for b-type joint capsule gene-specific multiplex PCR.%目的 应用多重PCR技术研究杭州地区儿童中分离的流感嗜血杆菌b型菌株的检出率.方法 以玻片凝集法的血清分型为金标准,以流感嗜血杆菌(Hi)荚膜编码基因(bexA)和b型特异性荚膜基因序列设计引物,应用多重PCR技术对流感嗜血杆菌菌株进行荚膜基因检测.结果 2001 - 2002年和2006-2007年分离的399株Hi临床株中,血清分型显示不可分型株297株,占74.44%,可分型株102株,占25.56%.b型仅1株,构成比0.98%.多重PCR检测显示:102株玻片凝集法可分型的菌株中,101株bex A PCR结果阳性,1株阴性;297株不定型株中2株bex A阳性.敏感度99.02%;特异度99.33%.Hib检测结果显示b型1株,占有荚膜菌株0.99%,与血清分型结果一致.结论bex A PCR联合针对b型特异性荚膜基因的多重PCR技术,对Hib检出率100%,克服了血清分型的弊端.

  8. 128株流感嗜血菌对抗菌药物耐药性分析%Drug resistance of 128 strains of haemophilus influenzae to antibiotics

    Institute of Scientific and Technical Information of China (English)

    张有忠; 都青; 李娜

    2011-01-01

    目的 对襄樊市中医院2008-2009年临床微生物室分离出的流感嗜血菌耐药性及耐药趋势进行回顾性分析,为指导临床合理用药提供科学依据;同时优化检测方法,提高检出率.方法 对128株流感嗜血菌进行β-内酰胺酶测定,采用K-B法进行体外药敏试验.结果 128例流感嗜血菌中,产β-内酰胺酶率为28.69%,对氨苄西林、氨苄西林/舒巴坦、头孢噻肟、磺胺甲噁唑/甲氧苄啶、头孢呋辛、环丙沙星、阿奇霉素、左氧氟沙星、氯霉素、亚胺培南的耐药率分别为47.66%、15.63%、28.91%、56.25%、27.34%、19.53%、19.53%、29.69%、46.09%、0.结论 流感嗜血菌对亚胺培南、氨苄西林/舒巴坦、环丙沙星、阿奇霉素的敏感性较高,临床医师可选择这些药物进行治疗.%OBJECTIVE To retrospectively analyze the drug resistance trend and drug resistance of Haemophiius influenzae isolated from clinical microorganism room in the Xiangfan Traditional Chinese Medicine H ospital in the whole year of 2009, to provide scientific data for the clinical on rational drug use and optimize the detection method and improve the detection rate. METHODS The determination of β-lactamase was performed on 128 strains of HI and drug sensitivity was tested in vitro by using the method of Kirty-Bauer. RESULTS In 128 strains of H. influenzae, the β-lactamase producing rate was 28.69%, the drug resistant rates to ampicillin, ampicillin sulbactam,cefotaxime, cotrimoxazole, cefuroxime, ciprofloxacin, azithromycin, levofloxacin, chloramphenicol and imipenem were 47. 66%, 15. 63%, 28. 91%, 56. 25%, 27. 34%, 19. 53%, 19. 53%, 29.69%, 46. 09% and 0,respectively. CONCLUSION H. influenzae is sensitive to imipenem, ampicillin/sulbactam, ciprofloxacin and azithromycin, the clinicians can choose these drugs for the treatment.

  9. Comparison of bacterial biomass and PRP production between different isolation of Haemophilus influenza type-b (Hib under different culture conditions

    Directory of Open Access Journals (Sweden)

    Hadi, A.

    2011-06-01

    Full Text Available Heamophilus influenzae type-b (Hib is a gram - negative pleomorphic bacterium that causes meningitis infections in children with the age of less than 5 years particularly in two years old infants. In the present study various isolates of Heamophilus influenzae from infants suspected to meningitis were collected, identified, characterized and were used throughout our experiments. Different culture media namely Brain Heart Infusion Broth (BHIB, Tryptic Soy Broth (TSB and GC medium Base (GCB which this medium was modified and prepared in our own laboratory, were compared to determine the highest bacterial yield. All media were added supplements 10mg/ml hemin & 0.01/ml IsovitaleX containing V factor. The bacterial yield for all Hib strains present in our laboratory were measured with an initial inoculums of 104 cfu per ml. The result showed very closed amount of biomass for all isolates however, GCB had slightly higher yield and ultimately we chose this medium for cultivation and extraction of capsular polysaccharide (CPS-b. In our laboratory we have adapted the PRP production according to our technical and instrumental availabilities which exists in our laboratories replacing ultra centrifugation to phenol chloroform to remove contaminants like endotoxin and proteins to the minimum level and also decreased the number of some chemical treatments while some steps were added in purification process. Our study showed although there were not significant differences between the PRP extract of the three isolates with average amount of 108 mg/lit, however, isolate ATCC10210 (ATF2 showed the highest amount with 192mg/lit and the least PRP was produced by isolate H.inf.1, with 16 mg/lit. It seems that the data can be categorized to a normal distribution with the mean of 106.4 and standard deviation of 6.25. This result was confirmed by one sample kolmogorov-Smirnov test, hence the PRP ≥192 mg/lit is statistically significant at a significant level of α =0

  10. Study of the radiation effects on nucleic acids and related compounds. Annual progress report, August 15, 1974--August 14, 1975. [X radiation, hamster cells, Haemophilus influenzae

    Energy Technology Data Exchange (ETDEWEB)

    Wang, S.Y.

    1975-01-01

    Interest is being centered on the chemical and physical nature of radiation-induced lesions to nucleic acids and their components. These investigations have revealed the enormous complexity of chemical events in these systems and the possible degradation of nucleic acids by strand breakage. Therefore, work in the ionizing radiation of DNA and its components has proceeded along a dual course. For chemical studies, our prime concern is the stepwise isolation and identification of the radiation products of derivatives of pyrimidines and the study of the actual mechanisms of their formation. For biological studies, H. influenzae cells, the Chinese hamster V79B-1 cell line, and the Dunn osteosarcoma lung colony system were used. During the last year, the method of synthesis of 5-hydroperoxymethyluracil (T/sub ..cap alpha../OOH) was greatly improved. Large-scale preparation of 5-hydroxy-6-hydroperoxy-5,6-dihydrothymine (T/sup 6/OOH) were carried out in order to study the action of T/sup 6/OOH on neighboring bases, glycosidic bond-breakage, cell mutagenesis, chromosomal aberrations, and possible synergistic effects on x radiation. These results allow one to relate radiobiological effects with radiation chemical changes in DNA.

  11. Meta-analysis of vaccine protective efficacy of Haemophilus influenzae type b vaccine%b型流感嗜血杆菌疫苗流行病学保护效果Meta分析

    Institute of Scientific and Technical Information of China (English)

    胡昱; 李倩; 戚小华; 陈雅萍; 陈恩富

    2013-01-01

    目的 评价b型流感嗜血杆菌疫苗(Hib)流行病学保护效果.方法 电子检索National Centerfor Biotechnology Information(NCBI,美国国家医学图书馆数据库)和Cochrane协作网图书馆等数据库,将有关接种Hib流行病学保护效果的研究纳入分析.使用RevMan5.1软件进行统计分析.结果 共纳入7篇文献,均为随机对照试验.Hib 对流感嗜血杆菌侵袭性疾病(Hib-D)保护效果(vE)为88%(95%CI:80% ~93%),对流感嗜血杆菌脑膜炎(HIB-M)的VE为84%(95%CI:74%~93%).结论 接种Hib可以降低因Hib感染所致侵袭性疾病和脑膜炎的发病率,具有良好的保护效果.%Objective To evaluate vaccine protective efficacy(VE) of Haemophilus influenzae type b vaccine (Hib).Methods The studies of VE for Hlib were included by searching National Center for Biotechnology Information and Cochrane Library,and meta-analysis was made by RevMan5.1.Results A total of 7 studies were included,all of which were random clinical trial.The VE of Hib against invasive Hib disease(Hib-D) was 88 % (95 % CI:80%-93 %).The VE of Hib against Hib Meningitis(Hib-M) was 84%(95% CI:74%-93%).Conchusions Hib inoculation can gain a good protection against Hib-D and Hib-M,and also can reduce the incidence rate of relative diseases.

  12. An antagonist of the platelet-activating factor receptor inhibits adherence of both nontypeable Haemophilus influenzae and Streptococcus pneumoniae to cultured human bronchial epithelial cells exposed to cigarette smoke

    Directory of Open Access Journals (Sweden)

    Shukla SD

    2016-07-01

    Full Text Available Shakti D Shukla,1,* Rory L Fairbairn,1,* David A Gell,1 Roger D Latham,1 Sukhwinder S Sohal,1,2 Eugene H Walters,1 Ronan F O’Toole11Breathe Well Centre, School of Medicine, Faculty of Health, University of Tasmania, Hobart, TAS, Australia; 2School of Health Sciences, Faculty of Health, University of Tasmania, Launceston, TAS, Australia*These authors contributed equally to this workBackground: COPD is emerging as the third largest cause of human mortality worldwide after heart disease and stroke. Tobacco smoking, the primary risk factor for the development of COPD, induces increased expression of platelet-activating factor receptor (PAFr in the lung epithelium. Nontypeable Haemophilus influenzae (NTHi and Streptococcus pneumoniae adhere to PAFr on the luminal surface of human respiratory tract epithelial cells.Objective: To investigate PAFr as a potential drug target for the prevention of infections caused by the main bacterial drivers of acute exacerbations in COPD patients, NTHi and S. pneumoniae.Methods: Human bronchial epithelial BEAS-2B cells were exposed to cigarette smoke extract (CSE. PAFr expression levels were determined using immunocytochemistry and quantitative polymerase chain reaction. The epithelial cells were challenged with either NTHi or S. pneumoniae labeled with fluorescein isothiocyanate, and bacterial adhesion was measured using immunofluorescence. The effect of a well-evaluated antagonist of PAFr, WEB-2086, on binding of the bacterial pathogens to BEAS-2B cells was then assessed. In silico studies of the tertiary structure of PAFr and the binding pocket for PAF and its antagonist WEB-2086 were undertaken.Results: PAFr expression by bronchial epithelial cells was upregulated by CSE, and significantly associated with increased bacterial adhesion. WEB-2086 reduced the epithelial adhesion by both NTHi and S. pneumoniae to levels observed for non-CSE-exposed cells. Furthermore, it was nontoxic toward the bronchial epithelial

  13. 巧克力琼脂对流感嗜血杆菌生长影响的探讨%Effects of chocolate agar medium on haemophilus influenza growth

    Institute of Scientific and Technical Information of China (English)

    林干; 卢勉飞; 容艳芬; 蔡芷荷; 严纪文; 吴清平

    2014-01-01

    目的 比较不同产地、不同动物血源配制的传统巧克力琼脂和改良巧克力琼脂对流感嗜血杆菌(Haemophilus influenzae)生长的影响.方法 对流感嗜血杆菌在不同产地、不同动物血源制备的传统巧克力琼脂平板与添加辅酶Ⅰ和混合动物血的改良巧克力琼脂平板的生长情况进行对比测试.结果 流感嗜血杆菌在不同动物血制备的巧克力琼脂上的生长差异有统计学意义(P<0.05),三种动物血源中以兔血最优、羊血次之,马血最差;若把兔血按1%、5%、10%的比例与羊血混合,流感嗜血杆菌的生长与在单独动物血制备的巧克力琼脂上的生长比较,差异无统计学意义(P>0.05);在不同动物血源的传统巧克力琼脂中添加辅酶Ⅰ 2 mg/100 ml后,流感嗜血杆菌的生长均明显增强,差异有统计学意义(P<0.05).结论 用兔血制备巧克力琼脂效果最优,添加辅酶Ⅰ可促进流感嗜血杆菌的生长.

  14. b型流感嗜血杆菌D蛋白的原核表达及纯化%Prokaryotic Expression and Purification of Protein D of Haemophilus influenza Type b

    Institute of Scientific and Technical Information of China (English)

    杜倩; 张华捷; 王一飞; 张庶民

    2011-01-01

    目的 克隆b型流感嗜血杆菌(Haemophlilus influenza type b,Hib)D蛋白(hpd)基因,原核表达并纯化重组hpd蛋白,为下一步开发以D蛋白为基础的结合疫苗奠定基础.方法 从Hib CMCC株基因组DNA中PCR扩增hpd基因片段,克隆入载体pET-30a(+),构建重组原核表达质粒pET-30a-hpd,转化入感受态大肠杆菌BL21(DE3),IPTG诱导表达.表达的重组蛋白经6mol/L尿素变性、DEAE阴离子交换柱纯化、透析复性后,采用Western blot法鉴定其反应原性.结果 重组表达质粒DET-30a-hpa经PCR及测序证明构建正确;表达的重组蛋白以包涵体形式存在,表达量约占菌体总蛋白的40%;经一步过柱纯化可得到纯度达95%左右的重组蛋白;纯化的重组蛋白可与Hib免疫小鼠制备的抗血清发生特异性反应.结论 已成功克隆了Hib hpd基因,并在大肠杆菌中表达了重组hpd蛋白,为下一步开发以D蛋白为基础的结合疫苗奠定了基础.%Objective To clone the hpcl gene encoding protein D of Haemophilus influenza type b (Hib), express in prokaryotic cells, purify the expressed product and lay a foundation of further development of protein D-based conjugate vaccine.Methods The hpd gene fragment was amplified from genomic DNA of Hib CMCC strain by PCR and cloned into vector pET-30a (+).The constructed recombinant plasmid pET-30a-hpd was transformed to competent E.coli BL21 (DE3) for expression under induction of IPTG.The expressed protein was de-naturalized with 6 mol/L urea, purified by DEAE anion exchange column chromatography, re-naturalized by dialysis and identified for reactogenicity by Western blot.Results PCR and sequencing proved that the recombinant plasmid pET-30a-hpd was constructed correctly.The expressed protein, in a form of inclusion body, contained about 40% of total somatic protein and reached a purity of about 95% after purification by one-step column chromatography.The purified recombinant protein showed specific reaction with the

  15. 北京市东城区1~12岁健康儿童流感嗜血杆菌携带状况调查%Carriage of Haemophilus influenzae among healthy children aged 1- 12 years in Dongcheng district, Beijing

    Institute of Scientific and Technical Information of China (English)

    邸明芝; 吴疆; 黄辉; 吕敏; 张玉萍; 周云; 李侠; 孙宏莉; 闫威; 翟力军

    2012-01-01

    Objective To understand the carriage status of Haemophilus influenzae among healthy children aged 1-12 years in Dongcheng district, Beijing. Methods Nasopharageal swabs were collected from 221 healthy children aged 1-12 years randomly selected in Dongcheng district every three months to isolate H. influenzae. Results The isolation rate of H. influenzae was 8.75% , and the cumulative carrying rate was 29.41%. The coverage of H. influenzae b conjugate vaccine was 5. 24%. Conclusion It is necessary to establish scientific surveillance system and develop appropriate H. influenzae vaccine.%目的 了解北京市东城区1~12岁健康儿童流感嗜血杆菌携带状况.方法 选取北京市东城区1 ~12岁健康儿童221名,每季度采集鼻咽拭子标本1次,进行流感嗜血杆菌培养、血清分型及耐药性检测.结果 东城区1~12岁健康儿童流感嗜血杆菌分离率为8.75%,累计携带率为29.41%,b型流感嗜血杆菌结合疫苗接种率为5.24%.结论 1~12岁儿童存在着一定流感嗜血杆菌疾病的风险,有必要建立科学的监测体系,研制出适合国情的流感嗜血杆菌疫苗.

  16. Immune Effect of a Booster with Domestic Haemophilus influenzae Type b PRP-TT Conjugate Vaccine%国产b型流感嗜血杆菌结合疫苗PRP-TT加强免疫效果评价

    Institute of Scientific and Technical Information of China (English)

    李亚南; 乔瑞洁; 李红; 刘佳; 王浩; 史晓玲; 叶强; 谢贵林

    2011-01-01

    目的 评价国产b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)结合疫苗PRP-TT的加强免疫效果.方法 分别用安儿宝和呵儿贝两种Hib结合疫苗,对在广西柳州市常住儿童进行3、4、5月龄3针免疫,1年后,进行第4剂加强免疫.分别于3针免疫1年后和加强免疫1个月后采血,分离血清,采用ELISA和体外杀菌实验(Serum bactericidal assay,SBA)分别检测免疫前后血清IsG抗体浓度和加强免疫后SBA抗体滴度.结果 安儿宝和呵儿贝疫苗3针免疫1年后,血清IgG抗体几何平均数浓度(Geometric mean concentrations,GMCs)分别为3.17 μg/ml和3.00 μg/ml,二者差异无统计学意义(P>0.05);其中,IgG抗体浓度在1.00μg/ml以上的血清所占的比例分别为79%(62/78)和76%(115/152),二者差异无统计学意义(P>0.05).加强免疫后1个月,两种疫苗免疫血清中IgG抗体GMCs分别为71.33μg/ml和65.35 μg/ml,二者差异无统计学意义(P>0.05);IgG抗体浓度均能100%达1.00 μg/ml以上.两种疫苗加强免疫后的血清IgG抗体浓度与各自加强免疫前比较,差异均有统计学意义(P0.05).两种疫苗免疫后血清IgG抗体浓度和SBA抗体滴度均具有相关性(r值分别为0.696和0.689,P<0.05).结论3针免疫1年后,两种疫苗免疫血清中IgG抗体仍保持较高的水平,第4剂加强免疫后,抗体水平迅速显著升高,100%达1.00μg/ml以上,且具有有效的体外杀菌功能.%Objective To evaluate the immune effect of a booster with domestic Haemophilus influenzae type b(Hib) PRP-TT conjugate vaccine. Methods The infants resided in Liuzhou City, Guangxi Zhuang Autonomous Region, China were inoculated with two kinds of PRP-TT conjugate vaccine (Anerbao and Heerbei) respectively, for 3 times at ages of 3, 4 and 5 months, and boosted with the same kind of vaccine 12 months later. The serum samples were collected 12 month after the 3rd dose and one month after the booster, and determined for IgG titers by ELISA and serum

  17. Rod Haemophilus a jeho laboratorní vyšetření v současné medicíně

    OpenAIRE

    KADLECOVÁ, Jana

    2014-01-01

    My bachelor´s thesis focuses on the most major pathogen of the genus named Haemophilus, and Haemophila influenzae. H. influenzae is an originator of more serious infections, mainly its type b, which is the most virulent organism, causing invasive diseases mainly with children and older people. It causes mainly meningitis and chronic obstructive pulmonary disease. In the theoretical part of my bachelor´s thesis I write about the genus Haemophilus and clinic significances of the most famous spe...

  18. b型流感嗜血杆菌疫苗及新型疫苗或联合疫苗的免疫原性评价方法%Evaluation on immunogenicity of Haemophilus influenzae type b (Hib) vaccine as well as novel and combined Hib vaccines

    Institute of Scientific and Technical Information of China (English)

    朱朗; 刘一非; 陆玮; 林纪胜; 高强

    2015-01-01

    b型流感嗜血杆菌(Haemophilus influenza type b,Hib)是全球范围内引起3个月到5岁龄儿童肺炎、脑膜炎和败血症等严重疾病的重要病原菌.接种疫苗是唯一有效的预防和控制Hib引起疾病的公共卫生措施,将Hib结合疫苗纳入儿童常规免疫程序的国家,Hib发病率和携带率均明显降低.本文现对Hib疫苗种类、免疫学特性、临床效力以及Hib新型疫苗或联合疫苗的免疫原性评价方法作一综述.

  19. Haemophilus segnis endocarditis

    DEFF Research Database (Denmark)

    Bangsborg, Jette Marie; Tvede, M; Skinhøj, P

    1988-01-01

    Haemophilus segnis is a rarely recognised commensal in the oropharynx. We wish to report the first published case of endocarditis caused by H. segnis. The patient, a 76-year-old female did not recover until after 2 courses of ampicillin given for a total of 57 days. In the second course of treatm......Haemophilus segnis is a rarely recognised commensal in the oropharynx. We wish to report the first published case of endocarditis caused by H. segnis. The patient, a 76-year-old female did not recover until after 2 courses of ampicillin given for a total of 57 days. In the second course...

  20. Development of an indirect competitive ELISA method for determination of Haemophilus influenzae type b polysaccharide%b型流感嗜血杆菌多糖间接竞争ELISA检测方法的建立

    Institute of Scientific and Technical Information of China (English)

    王明清; 管娇琼; 龙艺; 朱文勇; 廖国阳

    2015-01-01

    目的 建立b型流感嗜血杆菌(Haemophilus influenzae type b,Hib)多糖间接竞争ELISA检测方法,并进行验证及初步应用.方法 以Hib多糖一酪胺结合物(PRP-Ty)作为包被抗原,待测PRP作为竞争抗原,与Hib抗血清反应,建立间接竞争ELISA检测方法.采用棋盘法确定最佳包被多糖抗原浓度及Hib抗血清稀释倍数,绘制标准曲线,并确定最低检测限;对建立的方法进行批内和批间精密性验证;取Hib发酵液,在培养温度、接种量等因素相同的情况下,分别将培养液初始pH调至6.0±0.02、7.0±0.02和8.0±0.02,培养不同时间取样,采用建立的方法检测培养液内PRP含量.结果 PRP-Ty的最佳包被浓度为1.25 μg/ml,Hib抗血清的最佳稀释倍数为300倍.该方法检测的线性范围为6.25~ 200 ng/ ml,最低检测限为6.25 ng/ ml,回归方程为y=-23.12x+99.62,R2=0.996.采用该方法分别重复检测3次高浓度(200 ng/ml)和低浓度(20 ng/ml)PRP多糖的变异系数(CV)分别为1.608%和3.344%;采用该方法及传统化学检测法分别检测6次同一批Hib成品分装疫苗的多糖,该方法检测结果的CV值为5.75%,传统化学法检测结果的CV值为1.98%,均符合要求;该方法检测不同初始pH培养液培养的Hib发酵液,培养时间为8h左右时,PRP产量最高;当初始pH为8.0±0.02时,培养液内PRP含量增长最快,所达到的浓度最高.结论 建立了Hib多糖间接竞争ELISA检测方法,该方法灵敏度较高,精密性良好,可应用于Hib多糖含量的定量检测.