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Sample records for 19a streptococcus pneumoniae

  1. Effect of culture conditions on the structure of Streptococcus pneumoniae type 19A(57) capsular polysaccharide.

    Lee, C.J.; Fraser, B A; Boykins, R A; Li, J P

    1987-01-01

    The structural modifications and immunochemical activities of several Streptococcus pneumoniae type 19A polysaccharide (PS) preparations have been studied by sugar compositional analysis and immunodiffusion. The 19A PS preparations Lab-A-1 and Lab-A-3 and one PS isolated from 19A strain OB contained fucose (Fuc) and galactose (Gal) in addition to rhamnose (Rha) and glucose (Glc). In contrast, 19A PSs Lab-A-2 and Lab-B contained only Rha and Glc. Despite their different sugar compositions, the...

  2. Genome-wide dissection of globally emergent multi-drug resistant serotype 19A Streptococcus pneumoniae

    Dylan R Pillai; Shahinas, Dea; Buzina, Alla; Pollock, Remy A; Lau, Rachel; Khairnar, Krishna; Wong, Andrew; Farrell, David J.; Green, Karen; McGeer, Allison; Low, Donald E.

    2009-01-01

    Background Emergence of multi-drug resistant (MDR) serotype 19A Streptococcus pneumoniae (SPN) is well-documented but causal factors remain unclear. Canadian SPN isolates (1993-2008, n = 11,083) were serotyped and in vitro susceptibility tested. A subset of MDR 19A were multi-locus sequence typed (MLST) and representative isolates' whole genomes sequenced. Results MDR 19A increased in the post-PCV7 era while 19F, 6B, and 23F concurrently declined. MLST of MDR 19A (n = 97) revealed that sequen...

  3. Complete Genome Sequence of Streptococcus pneumoniae Serotype 19A, a Blood Clinical Isolate from Northeast Mexico

    Hinojosa-Robles, Rosa Maria; Barcenas-Walls, Jose Ramon; Vignau-Cantu, Armando; Barrera-Saldaña, Hugo A.

    2016-01-01

    We report here the draft genome sequence of a Streptococcus pneumoniae strain isolated in Monterrey, Mexico, MTY1662SN214, from a man with purpura fulminans. The strain belongs to the invasive and multidrug-resistant serogroup 19A, sequence type 320 (ST320). The draft genome sequence consists of 60 large contigs, a total of 2,069,474 bp, and has a G+C content of 39.7%. PMID:27034499

  4. Molecular epidemiology of serotype 19A Streptococcus pneumoniae isolated from children in Beijing, 1997-2006

    XUE Lian; YAO Kai-hu; YU Sang-jie; LIU Zun-jie; QIAN Jing; SHEN Xu-zhuang; YANG Yong-hong

    2011-01-01

    Background Despite the prevalence of Streptococcus pneumoniae serotype 19A, the molecular characteristics of this serotype are yet to be fully elucidated. The aim of this study was therefore to determine the homology of the serotype 19A in China.Methods Pulsed-field gel electrophoresis and multilocus sequence typing were done to these forty-nine serotype 19A isolates to investigate the relationship between the strains prevalent in Beijing and other regions. Results From 1997 to 2006, the percentage of serotype 19A isolates increased. The susceptibility rate to penicillin and amoxicillin decreased and the resistance rate to cefuroxime increased. ST320 was the most prevalent ST, followed by ST3546. There were six new STs identified in our study. The serotype 19A strains were classified into six different pulsed-field gel electrophoresis (PFGE) patterns. ST320, which was associated with two different PFGE patterns (A and D), accounted for 32 isolates, and ST3546, which was associated with two PFGE patterns (B and E), accounted for eightConclusions From 2003 onwards, ST320 was the most common ST and the rate of resistance to cefuroxime increased significantly. Further long-term surveys of Streptococcus pneumoniae serotype 19A are required to monitor ST prevalence and antimicrobial resistance in this important human pathogen.

  5. Genome-wide dissection of globally emergent multi-drug resistant serotype 19A Streptococcus pneumoniae

    Wong Andrew

    2009-12-01

    Full Text Available Abstract Background Emergence of multi-drug resistant (MDR serotype 19A Streptococcus pneumoniae (SPN is well-documented but causal factors remain unclear. Canadian SPN isolates (1993-2008, n = 11,083 were serotyped and in vitro susceptibility tested. A subset of MDR 19A were multi-locus sequence typed (MLST and representative isolates' whole genomes sequenced. Results MDR 19A increased in the post-PCV7 era while 19F, 6B, and 23F concurrently declined. MLST of MDR 19A (n = 97 revealed that sequence type (ST 320 predominated. ST320 was unique amongst MDR 19A in that its minimum inhibitory concentration (MIC values for penicillin, amoxicillin, ceftriaxone, and erythromycin were higher than for other ST present amongst post-PCV7 MDR 19A. DNA sequencing revealed that alleles at key drug resistance loci pbp2a, pbp2x, pbp2b, ermB, mefA/E, and tetM were conserved between pre-PCV7 ST 320 19F and post-PCV7 ST 320 19A most likely due to a capsule switch recombination event. A genome wide comparison of MDR 19A ST320 with MDR 19F ST320 identified 822 unique SNPs in 19A, 61 of which were present in antimicrobial resistance genes and 100 in virulence factors. Conclusions Our results suggest a complex genetic picture where high-level drug resistance, vaccine selection pressure, and SPN mutational events have created a "perfect storm" for the emergence of MDR 19A.

  6. Genome-wide dissection of globally emergent multi-drug resistant serotype 19A Streptococcus pneumoniae

    Wong Andrew; Khairnar Krishna; Lau Rachel; Pollock Remy A; Buzina Alla; Shahinas Dea; Pillai Dylan R; Farrell David J; Green Karen; McGeer Allison; Low Donald E

    2009-01-01

    Abstract Background Emergence of multi-drug resistant (MDR) serotype 19A Streptococcus pneumoniae (SPN) is well-documented but causal factors remain unclear. Canadian SPN isolates (1993-2008, n = 11,083) were serotyped and in vitro susceptibility tested. A subset of MDR 19A were multi-locus sequence typed (MLST) and representative isolates' whole genomes sequenced. Results MDR 19A increased in the post-PCV7 era while 19F, 6B, and 23F concurrently declined. MLST of MDR 19A (n = 97) revealed th...

  7. Identification of Streptococcus pneumoniae

    Kaijalainen, Tarja

    2006-01-01

    Objectives: Streptococcus pneumoniae, pneumococcus, is an importanthuman pathogen that causes both serious invasive infections, suchas septicaemia, meningitis and pneumonia, as well as mild upper respiratoryinfections. It also belongs to the normal nasopharyngeal microbialflora. The purpose of this study was to compare bacteriologicalphenotypic methods with genetechnological methods in the identificationof pneumococci, especially among suspect pneumococcal isolateslacking one or more typical ...

  8. Gene Regulation in Streptococcus pneumoniae: interplay between nutrition and virulence

    W.T. Hendriksen (Wouter)

    2010-01-01

    textabstractStreptococcus pneumoniae (the pneumococcus) is a Gram-positive bacterium, which belongs to the species of streptococci. Other pathogenic bacteria belonging to this class include Streptococcus pyogenes, Streptococcus agalactiae, Streptococcus suis, Streptococcus uberis, Streptococcus bovi

  9. Cloning of Minor Autolysin of Streptococcus Pneumoniae

    Mahboobi, R. (MSc); Fallah Mehrabadi, J. (PhD); MR Pourmand; R Mashhadi; Haddadi, A. (MD

    2015-01-01

    Background and Objective: Increased antibiotic resistant strains and inadequacy of current vaccines against pneumococcal infections necessitate the study of novel protein antigens. It seems that minor autolysin of Streptococcus pneumoniae may have antigenicity. Thus, we aimed at cloning its gene for the first time. Material and Methods: After DNA extraction of Streptococcus pneumoniae (ATCC 49619), Specific primers were designed for amplifying minor autolysin gene fragment, using PCR. The pur...

  10. Phenotypic Characterization of Streptococcus pneumoniae Biofilm Development

    Allegrucci, Magee; Hu, F.Z.; Shen, K.; J. Hayes; Ehrlich, Garth D.; Post, J Christopher; Sauer, Karin

    2006-01-01

    Streptococcus pneumoniae is among the most common pathogens associated with chronic otitis media with effusion, which has been hypothesized to be a biofilm disease. S. pneumoniae has been shown to form biofilms, however, little is known about the developmental process, the architecture, and the changes that occur upon biofilm development. In the current study we made use of a continuous-culture biofilm system to characterize biofilm development of 14 different S. pneumoniae strains representi...

  11. Emerging resistant serotypes of invasive Streptococcus pneumoniae

    Elshafie, Sittana; Taj-Aldeen, Saad J

    2016-01-01

    Background Streptococcus pneumoniae is the leading cause of meningitis and sepsis. The aim of the study was to analyze the distribution, vaccine serotype coverage, and antibiotic resistance of S. pneumoniae serotypes isolated from patients with invasive diseases, after the introduction of pneumococcal 7-valent conjugated vaccine (PCV-7). Methods A total of 134 isolates were collected from blood and cerebrospinal fluid specimens at Hamad Hospital during the period from 2005 to 2009. Isolate serotyping was done using the Quellung reaction. The prevaccination period was considered before 2005. Results The most common serotypes for all age groups were 3 (12.70%), 14 (11.90%), 1 (11.90%), 19A (9.00%), 9V (5.20%), 23F (5.20%), and 19F (4.50%). Coverage rates for infant conjugated vaccine (PCV-10), and the 13-valent conjugated vaccine (PCV-13) were 34.78%, 52.17%, and 78.26%, respectively. Coverage rates of these vaccines were 50%, 67.86%, and 75% for the 2–5 years age group; 27.12%, 40.68%, and 64.41% for the age group 6–64 years; and 25%, 33.33%, and 66.67% for the ≥65 years age group, respectively. The percentage of nonsusceptible isolates to penicillin, cefotaxime, and erythromycin were 43.86%, 16.66%, and 22.81%, respectively. Thirty-seven isolates (32.46%) were multidrug resistant (MDR) and belonged to serotypes 14, 19A, 19F, 23F, 1, 9V, 12F, 4, 6B, 3, and 15A. Compared to previous results before the introduction of PCV-7, there was a significant reduction in penicillin-nonsusceptable S. pneumoniae from 66.67% to 43.86%, and a slight insignificant reduction in erythromycin nonsusceptible strains from 27.60% to 22.8%, while there was a significant increase in cefotaxime nonsusceptible strains from 3.55% to 16.66%. Conclusion Invasive pneumococcal strains and the emergence of MDR serotypes is a global burden that must be addressed through multiple strategies, including vaccination, antibiotic stewardship, and continuous surveillance. PMID:27418844

  12. Dyrkningsnegativ Streptococcus pneumoniae endokarditis diagnosticeret med polymerasekaedereaktion

    Rasmussen, Rasmus Vedby; Kemp, Michael; Bangsborg, Jette Marie;

    2008-01-01

    A 60-year old man was admitted with sepsis and meningitis of unknown aetiology. Underlying aortic valve endocarditis was diagnosed by echocardiography and severe insufficiency led to aortic valve replacement. Application of broad-range PCR to cusp tissue revealed a DNA product, and a diagnosis of...... Streptococcus pneumoniae endocarditis was obtained by DNA sequencing....

  13. Novel Type of Streptococcus pneumoniae Causing Multidrug-Resistant Acute Otitis Media in Children

    Xu, Qingfu; Pichichero, Michael E.; Casey, Janet R.; Zeng, Mingtao

    2009-01-01

    After our recent discovery of a Streptococcus pneumoniae 19A “superbug” (Legacy strain) that is resistant to all Food and Drug Administration–approved antimicrobial drugs for treatment of acute otitis media (AOM) in children, other S. pneumoniae isolates from children with AOM were characterized by multilocus sequence typing (MLST). Among 40 isolates studied, 16 (40%) were serotype 19A, and 9 (23%) were resistant to multiple antimicrobial drugs. Two others had unreported sequence types (STs) ...

  14. Granzyme A impairs host defense during Streptococcus pneumoniae pneumonia.

    van den Boogaard, Florry E; van Gisbergen, Klaas P J M; Vernooy, Juanita H; Medema, Jan P; Roelofs, Joris J T H; van Zoelen, Marieke A D; Endeman, Henrik; Biesma, Douwe H; Boon, Louis; Van't Veer, Cornelis; de Vos, Alex F; van der Poll, Tom

    2016-08-01

    Streptococcus pneumoniae is the most common causative pathogen in community-acquired pneumonia (CAP). Granzyme A (GzmA) is a serine protease produced by a variety of cell types involved in the immune response. We sought to determine the role of GzmA on the host response during pneumococcal pneumonia. GzmA was measured in bronchoalveolar lavage fluid (BALF) harvested from CAP patients from the infected and contralateral uninfected side and in lung tissue slides from CAP patients and controls. In CAP patients, GzmA levels were increased in BALF obtained from the infected lung. Human lungs showed constitutive GzmA expression by both parenchymal and nonparenchymal cells. In an experimental setting, pneumonia was induced in wild-type (WT) and GzmA-deficient (GzmA(-/-)) mice by intranasal inoculation of S. pneumoniae In separate experiments, WT and GzmA(-/-) mice were treated with natural killer (NK) cell depleting antibodies. Upon infection with S. pneumoniae, GzmA(-/-) mice showed a better survival and lower bacterial counts in BALF and distant body sites compared with WT mice. Although NK cells showed strong GzmA expression, NK cell depletion did not influence bacterial loads in either WT or GzmA(-/-) mice. These results implicate that GzmA plays an unfavorable role in host defense during pneumococcal pneumonia by a mechanism that does not depend on NK cells. PMID:27343190

  15. [Thousand faces of Streptococcus pneumonia (pneumococcus) infections].

    Szabó, Bálint Gergely; Lénárt, Katalin Szidónia; Kádár, Béla; Gombos, Andrea; Dezsényi, Balázs; Szanka, Judit; Bobek, Ilona; Prinz, Gyula

    2015-11-01

    Incidence and mortality rates of infections caused by Streptococcus pneumoniae (pneumococcus) are high worldwide and in Hungary among paediatric as well as adult populations. Pneumococci account for 35-40% of community acquired adult pneumonias requiring hospitalization, while 25-30% of Streptococcus pneumoniae pneumonias are accompanied by bacteraemia. 5-7% of all infections are fatal but this rate is exponentially higher in high risk patients and elderly people. Mortality could reach 20% among patients with severe invasive pneumococcal infections. Complications may develop despite administration of adequate antibiotics. The authors summarize the epidemiology of pneumococcal infections, pathogenesis of non-invasive and invasive disease and present basic clinical aspects through demonstration of four cases. Early risk stratification, sampling of hemocultures, administration of antibiotics and wider application of active immunization could reduce the mortality of invasive disease. Anti-pneumococcal vaccination is advisable for adults of ≥50 years and high risk patients of ≥18 years who are susceptible to pneumococcal disease. PMID:26498896

  16. Lack of SOS repair in Streptococcus pneumoniae

    Wild-type strains of Streptococcus pneumoniae were non-mutable by UV radiation and thymidine starvation. Moreover, UV-irradiated pneumococcal ω2 phages were not reactivated in an irradiated host. This suggests that, in pneumococcus, there is no efficient inducible repair process similar to the SOS repair described in detail for E. coli. We also report that mutations cannot be induced by a process thought to be linked to competence during transformation with isogenic wild-type DNA either on wild-type strains or in strains in which the hex function of excision and repair of mismatched bases is inactive. (orig.)

  17. Clinical behavior of Streptococcus pneumoniae meningoencephalitis Comportamiento clinico y terapéutico de la meningoencefalitis por Streptococcus pneumoniae

    Raisa Bu-Coifiu Fanego; Dorta-Contreras, Alberto J; Bárbara Padilla-Docal; Martha O' Farril-Sanchez; Isabel Lopez-Hernandez

    2009-01-01

    OBJECTIVE: There was an increased number of cases of meningoencephalitis caused by Streptococcus pneumoniae, after the successful vaccination campaigns against Neisseria meningitidis and Haemophilus influenzae. This paper aims at describing the clinical characteristics, the laboratory findings, the complications, and the therapeutic management of these patients, who have been suffering from this disease since 1993 to 2006. METHOD: Twelve children with Streptococcus pneumoniae meningoencephali...

  18. Regulation of neuraminidase expression in Streptococcus pneumoniae

    Gualdi Luciana

    2012-09-01

    Full Text Available Abstract Background Sialic acid (N-acetylneuraminic acid; NeuNAc is one of the most important carbohydrates for Streptococcus pneumoniae due of its role as a carbon and energy source, receptor for adhesion and invasion and molecular signal for promotion of biofilm formation, nasopharyngeal carriage and invasion of the lung. Results In this work, NeuNAc and its metabolic derivative N-acetyl mannosamine (ManNAc were used to analyze regulatory mechanisms of the neuraminidase locus expression. Genomic and metabolic comparison to Streptococcus mitis, Streptococcus oralis, Streptococcus gordonii and Streptococcus sanguinis elucidates the metabolic association of the two amino sugars to different parts of the locus coding for the two main pneumococcal neuraminidases and confirms the substrate specificity of the respective ABC transporters. Quantitative gene expression analysis shows repression of the locus by glucose and induction of all predicted transcriptional units by ManNAc and NeuNAc, each inducing with higher efficiency the operon encoding for the transporter with higher specificity for the respective amino sugar. Cytofluorimetric analysis demonstrated enhanced surface exposure of NanA on pneumococci grown in NeuNAc and ManNAc and an activity assay allowed to quantify approximately twelve times as much neuraminidase activity on induced cells as opposed to glucose grown cells. Conclusions The present data increase the understanding of metabolic regulation of the nanAB locus and indicate that experiments aimed at the elucidation of the relevance of neuraminidases in pneumococcal virulence should possibly not be carried out on bacteria grown in glucose containing media.

  19. Case Report of Necrotizing Fasciitis Associated with Streptococcus pneumoniae

    Lei Jiao

    2016-01-01

    Full Text Available Necrotizing fasciitis, caused by Streptococcus pneumoniae, is an extremely rare and life-threatening bacterial soft tissue infection. We report a case of early necrotizing fasciitis associated with Streptococcus pneumoniae infection in a 26-year-old man who was immunocompromised with mixed connective tissue disease. The patient presented with acute, painful, erythematous, and edematous skin lesions of his right lower back, which rapidly progressed to the right knee. The patient underwent surgical exploration, and a diagnosis of necrotizing fasciitis was confirmed by pathological evidence of necrosis of the fascia and neutrophil infiltration in tissue biopsies. Cultures of fascial tissue biopsies and blood samples were positive for Streptococcus pneumoniae. To our knowledge, this is the first report of necrotizing fasciitis resulting from Streptococcus pneumoniae diagnosed at early phase; the patient recovered well without surgical debridement.

  20. Novel type of Streptococcus pneumoniae causing multidrug-resistant acute otitis media in children.

    Xu, Qingfu; Pichichero, Michael E; Casey, Janet R; Zeng, Mingtao

    2009-04-01

    After our recent discovery of a Streptococcus pneumoniae 19A "superbug" (Legacy strain) that is resistant to all Food and Drug Administration-approved antimicrobial drugs for treatment of acute otitis media (AOM) in children, other S. pneumoniae isolates from children with AOM were characterized by multilocus sequence typing (MLST). Among 40 isolates studied, 16 (40%) were serotype 19A, and 9 (23%) were resistant to multiple antimicrobial drugs. Two others had unreported sequence types (STs) that expressed the 19A capsule, and 8 (88%) of the 9 multidrug-resistant strains were serotype 19A, including the Legacy strain with the new ST-2722. In genetic relatedness, ST-2722 belonged to a cluster of reported strains of S. pneumoniae in which all strains had 6 of the same alleles as ST-156. The multidrug-resistant strains related to ST-156 expressed different capsular serotypes: 9V, 14, 11A, 15C, and 19F. PMID:19331730

  1. STREPTOCOCCUS PNEUMONIAE KERATITIS FOLLOWING LASER IN SITU KERATOMILEUSIS

    2011-01-01

    A 22-year-old man underwent bilateral laser in situ keratomileusis and developed Streptococcus pneumoniae keratitis after surgery.This complication occurred one day after the procedure in both eyes.Topical and systemic antibiotics were promptly administered.Bacterial culture was performed following corneal flap lift and scraping of the lesions.Afterwards,the therapeutic regimen was readjusted according to the culture results.Streptococcus pneumoniae was identified from the culture.Three months after the sur...

  2. Inhibition of transformation in Streptococcus pneumoniae by lysogeny.

    Moynet, D J; Tiraby, G J

    1980-01-01

    Streptococcus pneumoniae R6X was lysogenized with bacteriophage 304 isolated after mitomycin induction of an ungrouped alpha-hemolytic streptococcus. Lysogenized pneumococci lost their capacity to undergo genetic transformation: transformability was restored after cells were spontaneously cured of their prophage. Both lysogens and nonlysogens produced activator substance (competence factor), and both bound deoxyribonucleic acid in a deoxyribonuclease-resistant form. However, nonlysogens retai...

  3. Evolution of Streptococcus pneumoniae and its close commensal relatives

    Kilian, Mogens; Poulsen, Knud; Blomqvist, Trinelise;

    2008-01-01

    Streptococcus pneumoniae is a member of the Mitis group of streptococci which, according to 16S rRNA-sequence based phylogenetic reconstruction, includes 12 species. While other species of this group are considered prototypes of commensal bacteria, S. pneumoniae is among the most frequent microbial...... killers worldwide. Population genetic analysis of 118 strains, supported by demonstration of a distinct cell wall carbohydrate structure and competence pheromone sequence signature, shows that S. pneumoniae is one of several hundred evolutionary lineages forming a cluster separate from Streptococcus...... oralis and Streptococcus infantis. The remaining lineages of this distinct cluster are commensals previously collectively referred to as Streptococcus mitis and each represent separate species by traditional taxonomic standard. Virulence genes including the operon for capsule polysaccharide synthesis and...

  4. Nonencapsulated Streptococcus pneumoniae: Emergence and Pathogenesis.

    Keller, Lance E; Robinson, D Ashley; McDaniel, Larry S

    2016-01-01

    While significant protection from pneumococcal disease has been achieved by the use of polysaccharide and polysaccharide-protein conjugate vaccines, capsule-independent protection has been limited by serotype replacement along with disease caused by nonencapsulatedStreptococcus pneumoniae(NESp). NESp strains compose approximately 3% to 19% of asymptomatic carriage isolates and harbor multiple antibiotic resistance genes. Surface proteins unique to NESp enhance colonization and virulence despite the lack of a capsule even though the capsule has been thought to be required for pneumococcal pathogenesis. Genes for pneumococcal surface proteins replace the capsular polysaccharide (cps) locus in some NESp isolates, and these proteins aid in pneumococcal colonization and otitis media (OM). NESp strains have been isolated from patients with invasive and noninvasive pneumococcal disease, but noninvasive diseases, specifically, conjunctivitis (85%) and OM (8%), are of higher prevalence. Conjunctival strains are commonly of the so-called classical NESp lineages defined by multilocus sequence types (STs) ST344 and ST448, while sporadic NESp lineages such as ST1106 are more commonly isolated from patients with other diseases. Interestingly, sporadic lineages have significantly higher rates of recombination than classical lineages. Higher rates of recombination can lead to increased acquisition of antibiotic resistance and virulence factors, increasing the risk of disease and hindering treatment. NESp strains are a significant proportion of the pneumococcal population, can cause disease, and may be increasing in prevalence in the population due to effects on the pneumococcal niche caused by pneumococcal vaccines. Current vaccines are ineffective against NESp, and further research is necessary to develop vaccines effective against both encapsulated and nonencapsulated pneumococci. PMID:27006456

  5. Treatment of experimental pneumonia due to penicillin-resistant Streptococcus pneumoniae in immunocompetent rats.

    Gavaldà, J.; Capdevila, J A; Almirante, B; Otero, J.; Ruiz, I; Laguarda, M; Allende, H.; Crespo, E; Pigrau, C; Pahissa, A

    1997-01-01

    A model of pneumonia due to Streptococcus pneumoniae resistant to penicillin was developed in immunocompetent Wistar rats and was used to evaluate the efficacies of different doses of penicillin, cefotaxime, cefpirome, and vancomycin. Adult Wistar rats were challenged by intratracheal inoculation with 3 x 10(9) CFU of one strain of S. pneumoniae resistant to penicillin (MICs of penicillin, cefotaxime, cefpirome, and vancomycin, 2, 1, 0.5, and 0.5 microg/ml, respectively) suspended in brain he...

  6. Isolation of Streptococcus pneumoniae type 3 from equine species.

    Benson, C E; Sweeney, C.R.

    1984-01-01

    Streptococcus pneumoniae type 3 was isolated from seven tracheobronchial aspirates and one pleural tap of seven adult horses and one foal. There was no direct evidence in these horses that isolation of the pneumococcus was related to a specific disease syndrome. Presenting complaints included two horses with chronic cough, two horses with decreased exercise tolerance, one horse with exercise-induced pulmonary hemorrhage, and three horses with pneumonia. Antibiotic therapy resolved the primary...

  7. Susceptibility of Streptococcus pneumoniae to Fluoroquinolones in Canada▿

    Patel, Samir N.; McGeer, Allison; Melano, Roberto; Tyrrell, Gregory J.; Green, Karen; Dylan R Pillai; Low, Donald E.

    2011-01-01

    Ciprofloxacin, the first fluoroquinolone to be used to treat lower respiratory tract infections (LRTI), demonstrates poor potency against Streptococcus pneumoniae, and its use has been associated with the emergence of resistance. During the last decade, fluoroquinolones with enhanced in vitro activity against S. pneumoniae have replaced ciprofloxacin for the treatment of LRTI. Here, we analyzed the impact of more active fluoroquinolone usage on pneumococci by examining the fluoroquinolone usa...

  8. New Streptococcus pneumoniae Clones in Deceased Wild Chimpanzees▿

    Chi, Fang; Leider, Michaela; Leendertz, Fabian; Bergmann, Carina; Boesch, Christophe; Schenk, Svenja; Pauli, Georg; Ellerbrok, Heinz; Hakenbeck, Regine

    2007-01-01

    In wild chimpanzees in the Taï National Park, Côte d'Ivoire, sudden deaths which were preceded by respiratory problems had been observed since 1999. Two new clones of Streptococcus pneumoniae were identified in deceased apes on the basis of multilocus sequence typing analysis and ply, lytA, and pbp2x sequences. The findings suggest that virulent S. pneumoniae occurs in populations of wild chimpanzees with the potential to cause infections similar to those observed in humans.

  9. Deletion of a Cation Transporter Promotes Lysis in Streptococcus pneumoniae

    Neef, Jolanda; Andisi, Vahid Farshchi; Kim, Kwang S.; Kuipers, Oscar P.; Bijlsma, Jetta J. E.; Weiser, J.N.

    2011-01-01

    Streptococcus pneumoniae is a significant human pathogen which causes respiratory and serious invasive diseases. Mg(2+) is essential for life, and its concentration varies throughout the human body. Magnesium uptake plays an important role in the virulence of many bacterial pathogens. To study the M

  10. A new tool for transcription regulation in Streptococcus pneumoniae

    Nováková, Linda; Přenosilová, Lenka; Sušická, Zuzana; Janeček, Jiří; Novotná, Jana; Ulrych, Aleš; Branny, Pavel

    Washington : American Society for Microbiology, 2006, s. 72-73. ISBN 1-55581-400-X. [ASM Conferences Streptococcal Genetics. Saint Malo (FR), 18.06.2006-21.06.2006] Institutional research plan: CEZ:AV0Z50200510 Keywords : streptococcus pneumoniae * rnap Subject RIV: EE - Microbiology, Virology

  11. Outbreak of Streptococcus pneumoniae in a Psychiatric Unit

    2012-11-02

    Dr. Katherine Fleming-Dutra, an epidemiologist at CDC, discusses her investigation of a Streptococcus pneumoniae outbreak in a pediatric psychiatric unit.  Created: 11/2/2012 by National Center for Emerging and Zoonotic Infectious Diseases (NCEZID); National Center for Immunization and Respiratory Diseases (NCIRD).   Date Released: 11/5/2012.

  12. Clinical behavior of Streptococcus pneumoniae meningoencephalitis Comportamiento clinico y terapéutico de la meningoencefalitis por Streptococcus pneumoniae

    Raisa Bu-Coifiu Fanego

    2009-12-01

    Full Text Available OBJECTIVE: There was an increased number of cases of meningoencephalitis caused by Streptococcus pneumoniae, after the successful vaccination campaigns against Neisseria meningitidis and Haemophilus influenzae. This paper aims at describing the clinical characteristics, the laboratory findings, the complications, and the therapeutic management of these patients, who have been suffering from this disease since 1993 to 2006. METHOD: Twelve children with Streptococcus pneumoniae meningoencephalitis admitted to the pediatric hospital of San Miguel del Padron, City of Havana in this period were assessed. RESULTS: Children under one year are the most frequently affected. Septic shock and brain edema were the most severe complications. Three patients died, implying that this disease has a serious course. Early treatment of brain edema is very important to reduce mortality. The elective drugs for treatment of these cases of Streptococcus pneumoniae meningoencephalitis were vancomycin combined with cephalosporin, cefotaxime or ceftriaxone type. CONCLUSION: Patients with Streptococcus pneumoniae meningoencephalitis show clinical characteristics, complications, and sequels that are different to other bacterial meningoencephalitis, meaning that they could be helpful for physicians considering the differential diagnosis of meningoencephalitis.OBJETIVO: Existe un incremento de la meningoencefalitis producida por Streptococcus pneumoniae, después de las campañas exitosas de vacunación contra Neisseria meningitidis y Haemophilus influenzae. El objetivo de este trabajo es describir las caracteristicas clinicas, los hallazgos de laboratorio, las complicaciones y el manejo terapéutico de los pacientes que sufrieron esta enfermedad desde 1993 a 2006. MÉTODO: Se estudiaron doce niños con meningoencefalitis por Streptococcus pneumoniae ingresados en el Hospital Pediátrico de San Miguel del Padrón, Ciudad de La Habana en este periodo. RESULTADOS: Los ni

  13. PCR detection of Streptococcus pneumoniae and Haemophilus influenzae in pneumonia patients

    Abdeldaim, Guma M. K.

    2009-01-01

    PCR is a rapid, reproducible method for nucleic acid detection. However, this technology displays significant deficiencies when applied in clinical microbiology. This work’s aim was to improve current diagnostics and provide sensitive and quantitative real-time PCRs. Paper I describes the development of a sensitive and specific quantitative real-time PCR for the detection of Streptococcus pneumoniae, based on the Spn9802 DNA fragment. Applied to nasopharyngeal aspirates from 166 pneumonia pat...

  14. Increase in invasive Streptococcus pyogenes and Streptococcus pneumoniae infections in England, December 2010 to January 2011.

    Zakikhany, K; Degail, M A; Lamagni, T; Waight, P; Guy, R; Zhao, H; Efstratiou, A; Pebody, R; George, R; Ramsay, M

    2011-01-01

    Increases in invasive Streptococcus pyogenes and S. pneumoniae above the seasonally expected levels are currently being seen in England. Preliminary analyses suggest that the high level of influenza activity seen this winter may be contributing to an increased risk of concurrent invasive bacterial and influenza infections in children and young adults. PMID:21315057

  15. Correlations between computed tomography findings and clinical manifestations of Streptococcus pneumoniae pneumonia

    The aim of this study was to characterize the imaging features and compare computed tomography (CT) findings with clinical features of patients with Streptococcus pneumoniae pneumonia. We retrospectively reviewed 75 patients (44 men, 31 women; mean age 67 years) diagnosed with S. pneumoniae pneumonia who underwent chest CT scanning at our institution between January 2007 and August 2008. Diagnoses were based on detection of the S. pneumoniae antigen in urine. Chest CT scans revealed abnormalities in all patients. The predominant opacity patterns were an airspace pneumonia pattern (48%) and a bronchopneumonia pattern (48%), followed by an interstitial pneumonia pattern (4%). Consolidation was observed most frequently (84%) followed by ground glass opacity (82.7%), bronchial wall thickening (61.3%), and centrilobular nodules (49.3%). Airway dilatation (21.6%), pleural effusion (33.3%), lymphadenopathy (34.8%), and pulmonary emphysema (21.3%) were also observed. Pulmonary emphysema was significantly less frequent in patients with the bronchopneumonia pattern than in those without (p=0.007). The clinical features and CT findings did not differ significantly. CT image analysis showed that patients with S. pneumoniae pneumonia exhibited the bronchopneumonia and airspace pneumonia patterns with equal frequency. Bronchopneumonia pattern was less common in patients with preexisting emphysema. (author)

  16. Serotype-specific mortality from invasive Streptococcus pneumoniae disease revisited

    Martens, Pernille; Worm, Signe Westring; Lundgren, Bettina;

    2004-01-01

    with Streptococcus pneumoniae (pneumococci) causes significant morbidity and mortality. Case series and experimental data have shown that the capsular serotype is involved in the pathogenesis and a determinant of disease outcome. METHODS: Retrospective review of 464 cases of invasive disease among adults diagnosed......Serotype-specific mortality from invasive Streptococcus pneumoniae disease revisited.Martens P, Worm SW, Lundgren B, Konradsen HB, Benfield T. Department of Infectious Diseases 144, Hvidovre University Hospital, DK-2650 Hvidovre, Denmark. pernillemartens@yahoo.com BACKGROUND: Invasive infection...... between 1990 and 2001. Multivariate Cox proportional hazard analysis. RESULTS: After adjustment for other markers of disease severity, we found that infection with serotype 3 was associated with an increased relative risk (RR) of death of 2.54 (95% confidence interval (CI): 1.22-5.27), whereas infection...

  17. Novel Clones of Streptococcus pneumoniae Causing Invasive Disease in Malaysia

    Johanna M Jefferies; Mohd Yasim Mohd Yusof; Shamala Devi Sekaran; Clarke, Stuart C.

    2014-01-01

    Although Streptococcus pneumoniae is a leading cause of childhood disease in South East Asia, little has previously been reported regarding the epidemiology of invasive pneumococcal disease in Malaysia and very few studies have explored pneumococcal epidemiology using multilocus sequence typing (MLST). Here we describe serotype, multilocus sequence type (ST), and penicillin susceptibility of thirty pneumococcal invasive disease isolates received by the University of Malaya Medical Centre betw...

  18. Streptococcus pneumoniae biofilm formation and dispersion during colonization and disease

    Chao, Yashuan; Marks, Laura R.; Pettigrew, Melinda M.; Hakansson, Anders P.

    2015-01-01

    Streptococcus pneumoniae (the pneumococcus) is a common colonizer of the human nasopharynx. Despite a low rate of invasive disease, the high prevalence of colonization results in millions of infections and over one million deaths per year, mostly in individuals under the age of 5 and the elderly. Colonizing pneumococci form well-organized biofilm communities in the nasopharyngeal environment, but the specific role of biofilms and their interaction with the host during colonization and disease...

  19. Studies on emergence and spread of antibiotic resistant Streptococcus pneumoniae

    Karlsson, Diana

    2010-01-01

    Streptococcus pneumoniae is one of the major contributors to mortality and morbidity around the world. It causes a wide variety of diseases ranging from uncomplicated respiratory infections to life-threatening invasive infections such as meningitis and septicemia. In recent years, the effectiveness of antibiotic therapy has been hampered by the increasing rates of resistant pneumococci. As antibiotic resistance increases, there is a growing need for interventions that minimi...

  20. Identification and characterization of novel virulence factors in Streptococcus pneumoniae

    Wartha, Florian

    2008-01-01

    Streptococcus pneumoniae (the pneumococcus) is a major human pathogen with high morbidity and mortality worldwide. Increased antibiotic resistance and insufficient vaccination contribute to the re-emerging of this pathogen. Identifying novel virulence factors could lead to a better understanding of the pathology of pneumococcal disease and result in novel therapeutic approaches. We were able to show the presence of a surface-exposed pilus structure in pneumococci, made u...

  1. Streptococcus pneumoniae biofilm formation and dispersion during colonization and disease

    Yashuan eChao; Marks, Laura R.; Pettigrew, Melinda M.; Hakansson, Anders P.

    2015-01-01

    Streptococcus pneumoniae (the pneumococcus) is a common colonizer of the human nasopharynx. Despite a low rate of invasive disease, the high prevalence of colonization results in millions of infections and over 1 million deaths per year, mostly in individuals under the age of 5 and the elderly. Colonizing pneumococci form well-organized biofilm communities in the nasopharyngeal environment, but the specific role of biofilms and their interaction with the host during colonization and disease i...

  2. Structure of a conjugative element in Streptococcus pneumoniae

    Vijayakumar, M.N.; Priebe, S.D.; Guild, W.R.

    1986-06-01

    The authors have cloned and mapped a 69-kilobase (kb) region of the chromosome of Streptococcus pneumoniae DP1322, which carries the conjugative Omega(cat-tet) insertion from S. pneumoniae BM6001. This element proved to be 65.5 kb in size. Location of the junctions was facilitated by cloning a preferred target region from the wild-type strain Rx1 recipient genome. This target site was preferred by both the BM6001 element and the cat-erm-tet element from Streptococcus agalactiae B109. Within the BM6001 element cat and tet were separated by 30 kb, and cat was flanked by two copies of a sequence that was also present in the recipient strain Rx1 DNA. Another sequence at least 2.4 kb in size was found inside the BM6001 element and at two places in the Rx1 genome. Its role is unknown. The ends of the BM6001 element appear to be the same as those of the B109 element, both as seen after transfer to S. pneumoniae and as mapped by others in pDP5 after transposition in Streptococcus faecalis. No homology is seen between the ends of the BM6001 element and no evidence found suggesting that it ever circularizes.

  3. Streptococcus pneumoniae arginine synthesis genes promote growth and virulence in pneumococcal meningitis

    J.R. Piet; M. Geldhoff; B.D.C. van Schaik; M.C. Brouwer; M. Valls Seron; M.E. Jakobs; K. Schipper; Y. Pannekoek; A.H. Zwinderman; T. van der Poll; A.H.C. van Kampen; F. Baas; A van der Ende; D. van de Beek

    2014-01-01

    Streptococcus pneumoniae (pneumococcus) is a major human pathogen causing pneumonia, sepsis and bacterial meningitis. Using a clinical phenotype based approach with bacterial whole-genome sequencing we identified pneumococcal arginine biosynthesis genes to be associated with outcome in patients with

  4. Streptococcus pneumoniae carriage in the Gaza strip.

    Gili Regev-Yochay

    Full Text Available BACKGROUND: Pneumococcal infections cause major morbidity and mortality in developing countries. We report the epidemiology of S. pneumoniae carriage in a developing region, the Gaza strip, and evaluate the theoretical coverage of carriage strains by pneumococcal conjugate vaccines (PCVs. METHODOLOGY: In 2009 we conducted a cross-sectional survey of S. pneumoniae carriage in healthy children and their parents, living throughout the Gaza strip. Data were collected and nasopharyngeal swabs were obtained. Antibiotic susceptibilities were determined by Vitek-2 and serotypes by the Quellung reaction. PRINCIPAL FINDINGS: S. pneumoniae carriage was detected in 189/379 (50% of children and 30/376 (8% of parents. Carriage prevalence was highest in children <6 months of age (63%. Significant predictors for child carriage were number of household members and DCC attendance. The proportion of pediatric and adults isolates with serotypes included in PCV7 were 32% and 20% respectively, and 46% and 33% in PCV13 respectively. The most prominent non-vaccine serotypes (NVT were 35B, 15B/C and 23B. Penicillin-nonsusceptible strains were carried by 70% of carriers, penicillin-resistant strains (PRSP by 13% and Multi-drug-resistant (MDR by 30%. Of all PRSP isolates 54% belonged to serotypes included in PCV7 and 71% in the PCV13. Similarly, 59% and 73% of MDR-SP isolates, would theoretically be covered by PCV7 and PCV13, respectively. CONCLUSIONS: This study demonstrates that, PCV13-included strains were carried by 46% and 33% of pediatric and adult subjects respectively. In the absence of definitive data regarding the virulence of the NVT strains, it is difficult to predict the effect of PCVs on IPD in this region.

  5. Recombinant expression of Streptococcus pneumoniae capsular polysaccharides in Escherichia coli

    Kay, Emily J.; Yates, Laura E.; Terra, Vanessa S.; Cuccui, Jon; Wren, Brendan W.

    2016-01-01

    Currently, Streptococcus pneumoniae is responsible for over 14 million cases of pneumonia worldwide annually, and over 1 million deaths, the majority of them children. The major determinant for pathogenesis is a polysaccharide capsule that is variable and is used to distinguish strains based on their serotype. The capsule forms the basis of the pneumococcal polysaccharide vaccine (PPV23) that contains purified capsular polysaccharide from 23 serotypes, and the pneumococcal conjugate vaccine (PCV13), containing 13 common serotypes conjugated to CRM197 (mutant diphtheria toxin). Purified capsule from S. pneumoniae is required for pneumococcal conjugate vaccine production, and costs can be prohibitively high, limiting accessibility of the vaccine in low-income countries. In this study, we demonstrate the recombinant expression of the capsule-encoding locus from four different serotypes of S. pneumoniae within Escherichia coli. Furthermore, we attempt to identify the minimum set of genes necessary to reliably and efficiently express these capsules heterologously. These E. coli strains could be used to produce a supply of S. pneumoniae serotype-specific capsules without the need to culture pathogenic bacteria. Additionally, these strains could be applied to synthetic glycobiological applications: recombinant vaccine production using E. coli outer membrane vesicles or coupling to proteins using protein glycan coupling technology. PMID:27110302

  6. Mycoplasma pneumoniae and Streptococcus pneumoniae caused different microbial structure and correlation network in lung microbiota.

    Wang, Heping; Dai, Wenkui; Qiu, Chuangzhao; Li, Shuaicheng; Wang, Wenjian; Xu, Jianqiang; Li, Zhichuan; Wang, Hongmei; Li, Yuzheng; Yang, Zhenyu; Feng, Xin; Zhou, Qian; Han, Lijuan; Li, Yinhu; Zheng, Yuejie

    2016-06-01

    Pneumonia is one of the most serious diseases for children, with which lung microbiota are proved to be associated. We performed 16S rDNA analysis on broncho-alveolar lavage fluid (BALF) for 32 children with tracheomalacia (C group), pneumonia infected with Streptococcus pneumoniae (S. pneumoniae) (D1 group) or Mycoplasma pneumoniae (M. pneumoniae) (D2 group). Children with tracheomalacia held lower microbial diversity and accumulated Lactococcus (mean ± SD, 45.21%±5.07%, P value Mycoplasma (0.67%±1.25%, P value <0.01) respectively. Bacterial correlation in C group was mainly intermediated by Pseudomonas and Arthrobacter. Whilst, D1 group harbored simplest microbial correlation in three groups, and D2 group held the most complicated network, involving enriched Staphylococcus (0.26%±0.71%), Massilia (0.81%±2.42%). This will be of significance for understanding pneumonia incidence and progression more comprehensively, and discerning between bacterial infection and carriage. PMID:27293852

  7. Streptococcus pneumoniae resistentes a Penicilina en Lima - Perú

    Juan Fukuda Sharizawa; Juan Echevarria Zarate; Fernando Llanos Zavalaga; Augusto Yi Chu; Sara Palomino; Eduardo Gotuzzo Herencia; Carlos Carrillo Parodi

    1996-01-01

    Objetivo: Evaluar la prevalencia de Streptococcus pneumoniae resistente a penicilina (SPRP). Material y métodos: Se realizó un estudio transversal, multicéntrico, entre Noviembre de 1993 y Noviembre de 1994. Cultivos de sangre, líquido cefalorraquídeo (LCR), líquido pleural (LP), material de timpanocentesis y esputo fueron coleccionados de los laboratorios de microbiología de cuatro hospitales de Lima. Las pruebas de concentración inhibitoria mínima (CIM), fueron realizados usando métodos de ...

  8. Development of Streptococcus pneumoniae Vaccines Using Live Vectors

    Shifeng Wang

    2014-01-01

    Full Text Available Streptococcus pneumoniae still causes severe morbidity and mortality worldwide, especially in young children and the elderly. Much effort has been dedicated to developing protein-based universal vaccines to conquer the current shortcomings of capsular vaccines and capsular conjugate vaccines, such as serotype replacement, limited coverage and high costs. A recombinant live vector vaccine delivering protective antigens is a promising way to achieve this goal. In this review, we discuss the researches using live recombinant vaccines, mainly live attenuated Salmonella and lactic acid bacteria, to deliver pneumococcal antigens. We also discuss both the limitations and the future of these vaccines.

  9. Purification and preliminary crystallization of alanine racemase from Streptococcus pneumoniae

    Im Hookang

    2007-05-01

    Full Text Available Abstract Background Over the past fifteen years, antibiotic resistance in the Gram-positive opportunistic human pathogen Streptococcus pneumoniae has significantly increased. Clinical isolates from patients with community-acquired pneumonia or otitis media often display resistance to two or more antibiotics. Given the need for new therapeutics, we intend to investigate enzymes of cell wall biosynthesis as novel drug targets. Alanine racemase, a ubiquitous enzyme among bacteria and absent in humans, provides the essential cell wall precursor, D-alanine, which forms part of the tetrapeptide crosslinking the peptidoglycan layer. Results The alanine racemases gene from S. pneumoniae (alrSP was amplified by PCR and cloned and expressed in Escherichia coli. The 367 amino acid, 39854 Da dimeric enzyme was purified to electrophoretic homogeneity and preliminary crystals were obtained. Racemic activity was demonstrated through complementation of an alr auxotroph of E. coli growing on L-alanine. In an alanine racemases photometric assay, specific activities of 87.0 and 84.8 U mg-1 were determined for the conversion of D- to L-alanine and L- to D-alanine, respectively. Conclusion We have isolated and characterized the alanine racemase gene from the opportunistic human pathogen S. pneumoniae. The enzyme shows sufficient homology with other alanine racemases to allow its integration into our ongoing structure-based drug design project.

  10. Invasiveness of Serotypes and Clones of Streptococcus pneumoniae among Children in Finland

    William P Hanage; Kaijalainen, Tarja H.; Ritva K Syrjänen; Auranen, Kari; Leinonen, Maija; Mäkelä, P. Helena; Brian G. Spratt

    2005-01-01

    Streptococcus pneumoniae (the pneumococcus) causes diseases from otitis media to life-threatening invasive infection. The species is extremely antigenically and clonally diverse. We wished to determine odds ratios (ORs) for serotypes and clones of S. pneumoniae that cause invasive disease in Finland. A total of 224 isolates of S. pneumoniae from cases of invasive disease in children

  11. Commensal Streptococci Serve as a Reservoir for β-Lactam Resistance Genes in Streptococcus pneumoniae

    Jensen, Anders; Valdórsson, Oskar; Frimodt-Møller, Niels;

    2015-01-01

    Streptococcus pneumoniae is a leading cause of pneumonia, meningitis, septicemia, and middle ear infections. The incidence of S. pneumoniae isolates that are not susceptible to penicillin has risen worldwide and may be above 20% in some countries. Beta-lactam antibiotic resistance in pneumococci is...

  12. Influence of the blood bacterial load on the meningeal inflammatory response in Streptococcus pneumoniae meningitis

    Østergaard, C; O´Reilly, T; Brandt, C;

    2006-01-01

    was induced by intracisternal injection of approximately 1 x 10(6) CFU Streptococcus pneumoniae, type 3, and the 26 rabbits were either provided with approximately 1 x 10(6) CFU S. pneumoniae intravenously at 0 hour ("bacteraemic" rabbits, n = 9), immunized with paraformaldehyde-killed S. pneumoniae...

  13. Structure and Inhibition of Quorum Sensing Target from Streptococcus pneumoniae

    Singh,V.; Shi, W.; Almo, S.; Evans, G.; Furneaux, R.; Tyler, P.; Painter, G.; Lenz, D.; Mee, S.; et al.

    2006-01-01

    Streptococcus pneumoniae 5'-methylthioadenosine/S-adenosylhomocysteine hydrolase (MTAN) catalyzes the hydrolytic deadenylation of its substrates to form adenine and 5-methylthioribose or S-ribosylhomocysteine (SRH). MTAN is not found in mammals but is involved in bacterial quorum sensing. MTAN gene disruption affects the growth and pathogenicity of bacteria, making it a target for antibiotic design. Kinetic isotope effects and computational studies have established a dissociative S{sub N}1 transition state for Escherichia coli MTAN, and transition state analogues resembling the transition state are powerful inhibitors of the enzyme [Singh, V., Lee, J. L., Nunez, S., Howell, P. L., and Schramm, V. L. (2005) Biochemistry 44, 11647-11659]. The sequence of MTAN from S. pneumoniae is 40% identical to that of E. coli MTAN, but S. pneumoniae MTAN exhibits remarkably distinct kinetic and inhibitory properties. 5'-Methylthio-Immucillin-A (MT-ImmA) is a transition state analogue resembling an early S{sub N}1 transition state. It is a weak inhibitor of S. pneumoniae MTAN with a K{sub i} of 1.0 {mu}M. The X-ray structure of S. pneumoniae MTAN with MT-ImmA indicates a dimer with the methylthio group in a flexible hydrophobic pocket. Replacing the methyl group with phenyl (PhT-ImmA), tolyl (p-TolT-ImmA), or ethyl (EtT-ImmA) groups increases the affinity to give K{sub i} values of 335, 60, and 40 nM, respectively. DADMe-Immucillins are geometric and electrostatic mimics of a fully dissociated transition state and bind more tightly than Immucillins. MT-DADMe-Immucillin-A inhibits with a K{sub i} value of 24 nM, and replacing the 5'-methyl group with p-Cl-phenyl (p-Cl-PhT-DADMe-ImmA) gave a K{sub i}* value of 0.36 nM. The inhibitory potential of DADMe-Immucillins relative to the Immucillins supports a fully dissociated transition state structure for S. pneumoniae MTAN. Comparison of active site contacts in the X-ray crystal structures of E. coli and S. pneumoniae

  14. MULTILOCI SEQUESTERANT STRAINS OF STREPTOCOCCUS PNEUMONIAE ISOLATED FROM ELDERLY PATIENTS WITH COMMUNITY ACQUIRED PNEUMONIA

    A. V. Martynova

    2014-01-01

    Full Text Available Comuunity-acquired pneumonias in aged patients is the significant epidemiology problem for the public health of almost all the countries. Even more important the problem of microbiological monitoring and epidemiology surveillance for the S. pneumoniae strains as one of the ubiquitous pathogens causing as the community-acquired pneumonias as well the other infections of respiratory tract, what defines their different epidemiological meaning.Multilocus sequence typing is the perspective method of molecular epidemiological surveillance allowing to define the epidemiologically dangerous clones of the ubiquitous microorganisms as Streptococcus pneumomiae. The aim of our research was to conduct the multilocus sequence typing of pneumococci strains isolated in patients with community acquired pneumonias, bronchitis in aged patients.Materials and methods. There were taken 14 strains of S. pneumoniae, isolated in patients with community-acquired pneumonias (seven of them were multiresistant, eight strains were isolated from patients with the chronical onstructive lung diseases and four strains from carriers. Multilocus sequence typing was conduected according to method to M.C. Enright and B.G. Spratt (1998.Results. The strains, isolated in all populations were the related isolates of the species S. pneumoniae, the most of them had the unique genotype defining the sequence type for every strain. There were 6 strains of Taiwan 19F-14 genotype from 14 strains isolated in aged patients with community-acquired pneumonia. Among strains isolated from carriers there were prevailing the strai of R6 genotype.Conclusion. Multilocus sequence typing allows to identify the new genotypes and to prognose the appearing of epidemiologically dangerous strains with new peculiarities.

  15. Polyamine transporter in Streptococcus pneumoniae is essential for evading early innate immune responses in pneumococcal pneumonia.

    Rai, Aswathy N; Thornton, Justin A; Stokes, John; Sunesara, Imran; Swiatlo, Edwin; Nanduri, Bindu

    2016-01-01

    Streptococcus pneumoniae is the most common bacterial etiology of pneumococcal pneumonia in adults worldwide. Genomic plasticity, antibiotic resistance and extreme capsular antigenic variation complicates the design of effective therapeutic strategies. Polyamines are ubiquitous small cationic molecules necessary for full expression of pneumococcal virulence. Polyamine transport system is an attractive therapeutic target as it is highly conserved across pneumococcal serotypes. In this study, we compared an isogenic deletion strain of S. pneumoniae TIGR4 in polyamine transport operon (ΔpotABCD) with the wild type in a mouse model of pneumococcal pneumonia. Our results show that the wild type persists in mouse lung 24 h post infection while the mutant strain is cleared by host defense mechanisms. We show that intact potABCD is required for survival in the host by providing resistance to neutrophil killing. Comparative proteomics analysis of murine lungs infected with wild type and ΔpotABCD pneumococci identified expression of proteins that could confer protection to wild type strain and help establish infection. We identified ERM complex, PGLYRP1, PTPRC/CD45 and POSTN as new players in the pathogenesis of pneumococcal pneumonia. Additionally, we found that deficiency of polyamine transport leads to up regulation of the polyamine synthesis genes speE and cad in vitro. PMID:27247105

  16. 220D-F2 from Rubus ulmifolius Kills Streptococcus pneumoniae Planktonic Cells and Pneumococcal Biofilms

    Sharmila J Talekar; Sopio Chochua; Katie Nelson; Klugman, Keith P.; Quave, Cassandra L; Vidal, Jorge E.

    2014-01-01

    Streptococcus pneumoniae (pneumococcus) forms organized biofilms to persist in the human nasopharynx. This persistence allows the pneumococcus to produce severe diseases such as pneumonia, otitis media, bacteremia and meningitis that kill nearly a million children every year. While bacteremia and meningitis are mediated by planktonic pneumococci, biofilm structures are present during pneumonia and otitis media. The global emergence of S. pneumoniae strains resistant to most commonly prescribe...

  17. Protection against Streptococcus pneumoniae lung infection after nasopharyngeal colonization requires both humoral and cellular immune responses

    Wilson, R; Cohen, J.M.; Jose, R J; Vogel, C; Baxendale, H.; Brown, J. S.

    2014-01-01

    Streptococcus pneumoniae is a common cause of pneumonia and infective exacerbations of chronic lung disease, yet there are few data on how adaptive immunity can specifically prevent S. pneumoniae lung infection. We have used a murine model of nasopharyngeal colonization by the serotype 19F S. pneumoniae strain EF3030 followed by lung infection to investigate whether colonization protects against subsequent lung infection and the mechanisms involved. EF3030 colonization induced systemic and lo...

  18. NASOPHARYNGEAL CARRIAGE OF STREPTOCOCCUS PNEUMONIAE IN HEALTHY CHILDREN UNDER FIVE YEARS OLD IN CENTRAL LOMBOK REGENCY, INDONESIA.

    Hadinegoro, Sri Rezeki; Prayitno, Ari; Khoeri, Miftahuddin Majid; Djelantik, I Gusti Gede; Dewi, Nurhandini Eka; Indriyani, Sang Ayu Kompiang; Muttaqin, Zainul; Mudaliana, Siti; Safari, Dodi

    2016-05-01

    Colonization with Streptococcus pneumoniae is mostly symptomless, but can progress to respiratory or even systemic disease. We investigated nasopharyngeal carriage of Streptococcus pneumoniae in healthy children under five years of age in Central Lombok Regency, Indonesia. This cross sectional study was carried out in 2012 among 1,200 healthy children aged 2 to 60 months. A multiplex sequential PCR was employed to determine serotype of cultured S. pneumoniae and a disk diffusion method to assess susceptibility to antimicrobial drugs. S. pneumoniae was cultured from 554 children and the most frequent serotypes found were 6A/B (22% of pneumococcal strains), 19F (11%), 23F (10%), 15B/C (8%), and 19A and 14 (4% each). The majority of strains were still susceptible to clindamycin (97%), erythromycin (87%), chloramphenicol (81%), and penicillin (72%), with only 41% and 38% susceptible to tetracycline and sulfamethoxazole/trimethoprim, respectively. Continuous surveillance of S. pneumoniae carriage is important for future pneumococcal vaccination programs in Indonesia. PMID:27405132

  19. Novel clones of Streptococcus pneumoniae causing invasive disease in Malaysia.

    Johanna M Jefferies

    Full Text Available Although Streptococcus pneumoniae is a leading cause of childhood disease in South East Asia, little has previously been reported regarding the epidemiology of invasive pneumococcal disease in Malaysia and very few studies have explored pneumococcal epidemiology using multilocus sequence typing (MLST. Here we describe serotype, multilocus sequence type (ST, and penicillin susceptibility of thirty pneumococcal invasive disease isolates received by the University of Malaya Medical Centre between February 2000 and January 2007 and relate this to the serotypes included in current pneumococcal conjugate vaccines. A high level of diversity was observed; fourteen serotypes and 26 sequence types (ST, (11 of which were not previously described were detected from 30 isolates. Penicillin non-susceptible pneumococci accounted for 33% of isolates. The extent of molecular heterogeneity within carried and disease-causing Malaysian pneumococci remains unknown. Larger surveillance and epidemiological studies are now required in this region to provide robust evidence on which to base future vaccine policy.

  20. Genotyping and serotyping of macrolide and multidrug resistant Streptococcus pneumoniae isolated from carrier children

    S F Swedan

    2016-01-01

    Full Text Available Aims: Streptococcus pneumoniae, an opportunistic pathogen commonly carried asymptomatically in the nasopharynx of children, is associated with increasing rates of treatment failures due to a worldwide increase in drug resistance. We investigated the carriage of S. pneumoniae in children 5 years or younger, the identity of prevalent serotypes, the rates of resistance to macrolides and other antimicrobial agents and the genotypes responsible for macrolide resistance. Materials and Methods: Nasopharyngeal swabs were collected from 157 children under 5 years for cultural isolation of S. pneumoniae. Antibiogram of isolates  was determined using the disk diffusion test, and the minimal inhibitory concentration to macrolides was determined using the E-test. Isolate serotypes and macrolide resistance genes, erm(B and mef(E, were identified using multiplex polymerase chain reactions. Results: S. pneumoniae was recovered from 33.8% of children; 41.9% among males and 21.9% among females (P = 0.009. The highest carriage rate occurred among age groups 7-12 months and 49-60 months. Most frequent serotypes were 19F, 6A/B, 11A, 19A, 14 and 15B/C.  Resistance to macrolides was 60.4%. Resistance to oxacillin, trimethoprim/sulfamethoxazole and clindamycin was present among 90.6%, 54.7% and 32.1% of isolates, respectively. All isolates were susceptible to chloramphenicol, levofloxacin and vancomycin. Isolates resistant to one or more macrolide drugs were more likely to be multidrug resistant. Resistance to clindamycin or oxacillin coexisted with macrolide resistance. Among the erythromycin-resistant isolates, erm(B, mef(E and erm(B and mef(E genes were present at rates of 43.8%, 37.5% and 6.3%, respectively. Erm(B and mef(E were associated with very high level and moderate-to-high level resistance to macrolides, respectively. Conclusion: A significant proportion of children harboured macrolide and multidrug-resistant S. pneumoniae.

  1. Streptococcus pneumoniae-associated pneumonia complicated by purulent pericarditis: case series

    Objective: In the antibiotic era, purulent pericarditis is a rare entity. However, there are still reports of cases of the disease, which is associated with high mortality, and most such cases are attributed to delayed diagnosis. Approximately 40-50% of all cases of purulent pericarditis are caused by Gram-positive bacteria, Streptococcus pneumoniae in particular. Methods: We report four cases of pneumococcal pneumonia complicated by pericarditis, with different clinical features and levels of severity. Results: In three of the four cases, the main complication was cardiac tamponade. Microbiological screening (urinary antigen testing and pleural fluid culture) confirmed the diagnosis of severe pneumococcal pneumonia complicated by purulent pericarditis. Conclusions: In cases of pneumococcal pneumonia complicated by pericarditis, early diagnosis is of paramount importance to avoid severe hemodynamic compromise. The complications of acute pericarditis appear early in the clinical course of the infection. The most serious complications are cardiac tamponade and its consequences. Antibiotic therapy combined with pericardiocentesis drastically reduces the mortality associated with purulent pericarditis. (author)

  2. Streptococcus pneumoniae-associated pneumonia complicated by purulent pericarditis: case series

    Cilloniz, Catia; Torres, Antoni [Servicio de Neumologia, Hospital Clinic de Barcelona, Ciber de Enfermedades Respiratorias (CIBERES), Instituto de Investigacion Biomedica Agusti Pi i Sunyer, Universidad de Barcelona (Spain); Rangel, Ernesto [Facultad de Medicina, Universidad Autonoma de Nayarit, Tepic (Mexico); Barlascini, Cornelius [Servizio di Igiene e Sanita Pubblica, Ospedale Generale di Sestri Levante, Sestri Levante (Italy); Piroddi, Ines Maria Grazia; Nicolini, Antonello, E-mail: antonellonicolini@gmail.com [Servizio di Pneumologia, Ospedale Generale di Sestri Levante, Sestri Levante (Italy)

    2015-07-15

    Objective: In the antibiotic era, purulent pericarditis is a rare entity. However, there are still reports of cases of the disease, which is associated with high mortality, and most such cases are attributed to delayed diagnosis. Approximately 40-50% of all cases of purulent pericarditis are caused by Gram-positive bacteria, Streptococcus pneumoniae in particular. Methods: We report four cases of pneumococcal pneumonia complicated by pericarditis, with different clinical features and levels of severity. Results: In three of the four cases, the main complication was cardiac tamponade. Microbiological screening (urinary antigen testing and pleural fluid culture) confirmed the diagnosis of severe pneumococcal pneumonia complicated by purulent pericarditis. Conclusions: In cases of pneumococcal pneumonia complicated by pericarditis, early diagnosis is of paramount importance to avoid severe hemodynamic compromise. The complications of acute pericarditis appear early in the clinical course of the infection. The most serious complications are cardiac tamponade and its consequences. Antibiotic therapy combined with pericardiocentesis drastically reduces the mortality associated with purulent pericarditis. (author)

  3. Conjugal mobilization of the mega element carrying mef(E) from Streptococcus salivarius to Streptococcus pneumoniae.

    Santagati, Maria; Lupo, Agnese; Scillato, Marina; Di Martino, Andrea; Stefani, Stefania

    2009-01-01

    We report the isolation and characterization of an unusual strain of Streptococcus salivarius, 3C30, displaying both the macrolide-lincosamide-streptogramin B and the tetracycline resistance phenotypes. It harbours the mef(E), erm(B), and tet(M) genes carried by different genetic elements. The genetic element carrying mef(E), named mega, was investigated by long PCR and sequencing, while the presence of the Tn3872-like element, carrying tet(M) and erm(B), was demonstrated by sequencing of both the int-xis-Tn and the fragment between the two resistance genes. In strain 3C30 the mega element is 5388 bp in size and its nucleotide sequence is identical to that of the element described previously in S. salivarius, with the exception of a 912 bp deletion at the left end. The composite Tn3872-like element appeared to be nonconjugative while the mega element was transferred by conjugation to Streptococcus pneumoniae. It was, however, impossible to transfer it again from these transconjugants to other strains. In addition, only in the 3C30 strain did mega form circular structures, as identified by real-time PCR. In conclusion, we found a clinical strain of S. salivarius carrying both mega and Tn3872-like genetic elements. Mega is transferable by conjugation to S. pneumoniae but it is not transferable again from the transconjugants, suggesting a possible mobilization by recombinases of the coresident Tn3872-like transposon. PMID:19025575

  4. Serotype distribution and antimicrobial resistance of Streptococcus pneumoniae causing noninvasive diseases in a Children's Hospital, Shanghai

    Fen Pan

    2015-04-01

    Full Text Available Background:Streptococcus pneumoniae, which cause noninvasive pneumococcal diseases, severely impair children's health. This study analyzed serotype distribution and antimicrobial resistance of S. pneumoniae from January 2012 to December 2012 in a Children's Hospital, Shanghai.Methods:A total of 328 pneumococcal isolates were serotyped by multiplex sequential PCR and/or capsule-quellung reaction. The minimum inhibitory concentrations for 11 antimi- crobial agents were determined by broth microdilution method.Results:Among 328 strains, 19F (36.3%, 19A (13.4%, 6A (11.9%, 23F (11.0%, 14 (5.8%, 6B (5.2%, and 15B/C (4.3% were the most common serotypes. The coverage rates of 7-, 10-, and 13-valent conjugate vaccines (PCV7, PCV10, and PCV13 were 58.2%, 58.2%, and 84.1%, respectively. Out of the isolates, 26 (7.9% strains were penicillin resistant. Most of the strains displayed high resistance rate to macrolides (98.5% to erythromycin, 97.9% to azithromycin, and 97.0% to clindamycin.Conclusions:The potential coverage of PCV13 is higher than PCV7 and PCV10 because of the emergence of 19A and there should be long-term and systematic surveillance for non-vaccine serotypes.

  5. Characterization of erythromycin-resistant Streptococcus pneumoniae isolates causing invasive diseases in Chinese children

    MA Xiang; YAO Kai-hu; XIE Gui-lin; ZHENG YUE-jie; WANG Chuan-qing; SHANG Yun-xiao; WANG Hui-yun

    2013-01-01

    Background Erythromycin-resistant Streptococcus pneumoniae isolates that causing invasive pneumococcal diseases (IPD) in Chinese children remain uncharacterized.This study aims to identify the resistance genes associated with erythromycin resistance and to determine the genetic relationships of IPD isolates in Chinese children.Methods A total of 171 S.pneumoniae strains were isolated from 11 medical centers in China from 2006 to 2008.All the isolates were characterized via serotyping and antibiotic susceptibility determination.The erythromycin-resistant isolates were further characterized via ermB and mefA gene detection,multi-locus sequence typing analysis,and pulsed-field gel electrophoresis.Results A total of 164 (95.9%) isolates showed resistance to erythromycin,of which 162 strains with high high-level resistance (MIC ≥ 256 μg/ml).A total of 104 (63.4%) isolates carry the ermB gene alone,whereas 59 (36.0%) harbor both ermB and mefA genes.Of the 59 strains,54 were of serotypes 19A and 19F and were identified as highly clonal and related to the Taiwan19F-14 clone.Conclusions The erythromycin resistance rate in IPD isolates is significantly high and is predominantly mediated by the ermB gene.Isolates that carry both ermB and mefA genes are predominantly of serotypes 19A and 19F.

  6. Streptococcus pneumoniae from Palestinian nasopharyngeal carriers: serotype distribution and antimicrobial resistance.

    Abedelmajeed Nasereddin

    Full Text Available Infections of Streptococcus pneumoniae in children can be prevented by vaccination; left untreated, they cause high morbidity and fatalities. This study aimed at determining the nasopharyngeal carrier rates, serotype distribution and antimicrobial resistance patterns of S. pneumoniae in healthy Palestinian children under age two prior to the full introduction of the pneumococcal 7-valent conjugate vaccine (PCV7, which was originally introduced into Palestine in a pilot trial in September, 2010. In a cross sectional study, nasopharyngeal specimens were collected from 397 healthy children from different Palestinian districts between the beginning of November 2012 to the end of January 2013. Samples were inoculated into blood agar and suspected colonies were examined by amplifying the pneumococcal-specific autolysin gene using a real-time PCR. Serotypes were identified by a PCR that incorporated different sets of specific primers. Antimicrobial susceptibility was measured by disk diffusion and MIC methods. The resulting carrier rate of Streptococcus pneumoniae was 55.7% (221/397. The main serotypes were PCV7 serotypes 19F (12.2%, 23F (9.0%, 6B (8.6% and 14 (4% and PCV13 serotypes 6A (13.6% and 19A (4.1%. Notably, serotype 6A, not included in the pilot trial (PCV7 vaccine, was the most prevalent. Resistance to more than two drugs was observed for bacteria from 34.1% of the children (72/211 while 22.3% (47/211 carried bacteria were susceptible to all tested antibiotics. All the isolates were sensitive to cefotaxime and vancomycin. Any or all of these might impinge on the type and efficacy of the pneumococcal conjugate vaccines and antibiotics to be used for prevention and treatment of pneumococcal disease in the country.

  7. Time to positivity in blood cultures of adults with Streptococcus pneumoniae bacteremia

    Ansorena Luis; Garrido Jose; Rodríguez-Lera María; Peralta Galo; Roiz María

    2006-01-01

    Abstract Background previous studies have established that bacterial blood concentration is related with clinical outcome. Time to positivity of blood cultures (TTP) has relationship with bacterial blood concentration and could be related with prognosis. As there is scarce information about the usefulness of TTP, we study the relationship of TTP with clinical parameters in patients with Streptococcus pneumoniae bacteremia. Methods TTP of all cases of Streptococcus pneumoniae bacteremia, detec...

  8. AdcAII of Streptococcus pneumoniae Affects Pneumococcal Invasiveness.

    Lindsey R Brown

    Full Text Available Across bacterial species, metal binding proteins can serve functions in pathogenesis in addition to regulating metal homeostasis. We have compared and contrasted the activities of zinc (Zn2+-binding lipoproteins AdcA and AdcAII in the Streptococcus pneumoniae TIGR4 background. Exposure to Zn2+-limiting conditions resulted in delayed growth in a strain lacking AdcAII (ΔAdcAII when compared to wild type bacteria or a mutant lacking AdcA (ΔAdcA. AdcAII failed to interact with the extracellular matrix protein laminin despite homology to laminin-binding proteins of related streptococci. Deletion of AdcA or AdcAII led to significantly increased invasion of A549 human lung epithelial cells and a trend toward increased invasion in vivo. Loss of AdcAII, but not AdcA, was shown to negatively impact early colonization of the nasopharynx. Our findings suggest that expression of AdcAII affects invasiveness of S. pneumoniae in response to available Zn2+ concentrations.

  9. Serotypes of Streptococcus pneumoniae causing major pneumococcal infections

    Yu. V. Lobzin

    2014-09-01

    Full Text Available First in Russia prospective non-interventional hospital-based study on Streptococcus pneumoniae serotypes causing meningitis and acute otitis media (AOM in children and community-acquired pneumonia (CAP in children and adults, as well as serotype coverage by pneumococcal conjugate vaccines (PCV’s of different composition has been conducted. Serotypes 19F, 14 and serogroup 6 are the leading in meningitis; serotype coverage is 70,6% for PCV7, and 76,5% – for PCV10 and PCV13. Among S. pneumoniae serotypes causing AOM 19F, 3, 23F and serogroup 6 have been the most prevalent in Saint Petersburg. PCV7 and PCV10 provide equal serotypes coverage in AOM – 63,2% among children 0–2 years old, and 32,5% among children 5–17 years old. PCV13 covers up to 79% of serotypes in infants. In CAP PCV7 and PCV10 provide 57,1% serotype coverage in children and 56,1% – in adults. Serotype coverage in CAP for PCV13 has been 14,3% and 34,5% higher for children and adults, correspondingly. Obtained data supports PCV inclusion in children immunization program in Saint Petersburg, whereas PCV13 provides the broadest serotype coverage. In the course PCV’s implementation continued pneumococcal infection surveillance is advisable.

  10. Severe respiratory failure due to co-infection with human metapneumovirus and Streptococcus pneumoniae

    Masafumi Seki; Hisao Yoshida; Kazuyoshi Gotoh; Nobuyuki Hamada; Daisuke Motooka; Shota Nakamura; Norihisa Yamamoto; Shigeto Hamaguchi; Yukihiro Akeda; Hiroshi Watanabe; Tetsuya Iida; Kazunori Tomono

    2014-01-01

    A 64-year-old male patient was admitted with respiratory failure, although chest X-rays revealed only mild bronchiolitis. Streptococcus pneumoniae, which usually presents as massive lobular pneumonia, was isolated from sputum, however, pan-pathogen screening using a next-generation sequencer also detected human metapneumovirus genome fragments.

  11. Characterization and transfer studies of macrolide resistance genes in Streptococcus pneumoniae from Denmark

    Nielsen, Karen L; Hammerum, Anette M; Lambertsen, Lotte M; Lester, Camilla H; Arpi, Magnus; Knudsen, Jenny D; Stegger, Marc; Tolker-Nielsen, Tim; Frimodt-Møller, Niels

    2010-01-01

    Over the last decade, erythromycin resistance has been increasing in frequency in Streptococcus pneumoniae in Denmark. In the present study, 49 non-related erythromycin-resistant S. pneumoniae isolates from invasive sites and 20 isolates from non-invasive sites were collected; antimicrobial...

  12. R-roscovitine Reduces Lung Inflammation Induced by Lipoteichoic Acid and Streptococcus pneumoniae

    Hoogendijk, Arie J.; Roelofs, Joris J. T. H.; Duitman, JanWillem; van Lieshout, Miriam H. P.; Blok, Dana C; van der Poll, Tom; Wieland, Catharina W.

    2012-01-01

    Bacterial pneumonia remains associated with high morbidity and mortality. The gram-positive pathogen Streptococcus pneumoniae is the most common cause of community-acquired pneumonia. Lipoteichoic acid (LTA) is an important proinflammatory component of the gram-positive bacterial cell wall. R-roscovitine, a purine analog, is a potent cyclin-dependent kinase (CDK)-1, −2, −5 and −7 inhibitor that has the ability to inhibit the cell cycle and to induce polymorphonuclear cell (PMN) apoptosis. We ...

  13. Commensal Streptococci Serve as a Reservoir for β-Lactam Resistance Genes in Streptococcus pneumoniae

    Jensen, Anders; Valdórsson, Oskar; Frimodt-Møller, Niels; Hollingshead, Susan; Kilian, Mogens

    2015-01-01

    Streptococcus pneumoniae is a leading cause of pneumonia, meningitis, septicemia, and middle ear infections. The incidence of S. pneumoniae isolates that are not susceptible to penicillin has risen worldwide and may be above 20% in some countries. Beta-lactam antibiotic resistance in pneumococci is associated with significant sequence polymorphism in penicillin-binding proteins (PBPs). Commensal streptococci, especially S. mitis and S. oralis, have been identified as putative donors of mutate...

  14. Influenza Virus Infection Decreases Tracheal Mucociliary Velocity and Clearance of Streptococcus pneumoniae

    Pittet, Lynnelle A.; Hall-Stoodley, Luanne; Rutkowski, Melanie R.; Harmsen, Allen G.

    2009-01-01

    Influenza virus infections increase susceptibility to secondary bacterial infections, such as pneumococcal pneumonia, resulting in increased morbidity and mortality. Influenza-induced tissue damage is hypothesized to increase susceptibility to Streptococcus pneumoniae infection by increasing adherence to the respiratory epithelium. Using a mouse model of influenza infection followed by S. pneumoniae infection, we found that an influenza infection does not increase the number of pneumococci in...

  15. Invasive Streptococcus pneumoniae infection causing hemolytic uremic syndrome in children: Two recent cases

    Vanderkooi, Otto G; Kellner, James D; Wade, Andrew W.; Jadavji, Tajdin; Midgley, Julian P; Louie, Thomas; Tyrrell, Gregory J.

    2003-01-01

    INTRODUCTION: Streptococcus pneumoniae is an uncommon cause of hemolytic uremic syndrome (HUS) with a unique pathophysiology that differs from Shiga toxin-related HUS.METHODS: Case descriptions for each patient are provided. Each strain of S pneumoniae was subjected to a pulsed-field gel electrophoresis (PFGE) analysis, Shiga toxin assay and polymerase chain reaction to detect Shiga toxin genes. A review of the current literature was conducted.CASE PRESENTATIONS: Two patients with S pneumonia...

  16. Complete Genome Sequence of Streptococcus pneumoniae Strain A026, a Clinical Multidrug-Resistant Isolate Carrying Tn2010

    Sui, Zhihai; Zhou, Wenqing; Yao, Kaihu; Liu, Li; Zhang, Gang; Yang, Yonghong; Feng, Jie

    2013-01-01

    Streptococcus pneumoniae is a primary cause of bacterial infection in humans. Here, we present the complete genome sequence of S. pneumoniae strain A026, which is a multidrug-resistant strain isolated from cerebrospinal fluid.

  17. Streptococcus pneumoniae biofilm formation and dispersion during colonization and disease

    Yashuan eChao

    2015-01-01

    Full Text Available Streptococcus pneumoniae (the pneumococcus is a common colonizer of the human nasopharynx. Despite a low rate of invasive disease, the high prevalence of colonization results in millions of infections and over 1 million deaths per year, mostly in individuals under the age of 5 and the elderly. Colonizing pneumococci form well-organized biofilm communities in the nasopharyngeal environment, but the specific role of biofilms and their interaction with the host during colonization and disease is not yet clear. Pneumococci in biofilms are highly resistant to antimicrobial agents and this phenotype can be recapitulated when pneumococci are grown on respiratory epithelial cells under conditions found in the nasopharyngeal environment. Pneumococcal biofilms display lower levels of virulence in vivo and provide an optimal environment for increased genetic exchange both in vitro and in vivo, with increased natural transformation seen during co-colonization with multiple strains. Biofilms have also been detected on mucosal surfaces during pneumonia and middle ear infection, although the role of these biofilms in the disease process is debated. Recent studies have shown that changes in the nasopharyngeal environment caused by concomitant virus infection, changes in the microflora, inflammation, or other host assaults trigger active release of pneumococci from biofilms. These dispersed bacteria have distinct phenotypic properties and transcriptional profiles different from both biofilm and broth-grown, planktonic bacteria, resulting in a significantly increased virulence in vivo.In this review we discuss the properties of pneumococcal biofilms, the role of biofilm formation during pneumococcal colonization, including their propensity for increased ability to exchange genetic material, as well as mechanisms involved in transition from asymptomatic biofilm colonization to dissemination and disease of otherwise sterile sites. Greater understanding of

  18. Amoxicillin Is Effective against Penicillin-Resistant Streptococcus pneumoniae Strains in a Mouse Pneumonia Model Simulating Human Pharmacokinetics▿

    Abgueguen, Pierre; Azoulay-Dupuis, Esther; Noel, Violaine; Moine, Pierre; Rieux, Veronique; Fantin, Bruno; Bedos, Jean-Pierre

    2006-01-01

    High-dose oral amoxicillin (3 g/day) is the recommended empirical outpatient treatment of community-acquired pneumonia (CAP) in many European guidelines. To investigate the clinical efficacy of this treatment in CAP caused by Streptococcus pneumoniae strains with MICs of amoxicillin ≥2 μg/ml, we used a lethal bacteremic pneumonia model in leukopenic female Swiss mice with induced renal failure to replicate amoxicillin kinetics in humans given 1 g/8 h orally. Amoxicillin (15 mg/kg of body weig...

  19. The Use of Microarray Technology for the Analysis of Streptococcus Pneumoniae

    Timothy J. Mitchell

    2006-04-01

    Full Text Available Streptococcus pneumoniae is an important human pathogen associated with pneumonia, septicaemia, meningitis and otitis media. It is estimated to result in over 3 million child deaths worldwide every year and an even greater number of deaths among the elderly. Prior to the complete sequencing of the genomes of S. pneumoniae TIGR4 (serotype 4 and S. pneumoniae R6 (serotype 2, we designed a custom miniarray consisting of 497 pneumococcal genes. The overall objectives of our microarray investigations were, first, to assess the genetic diversity between different S. pneumoniae serotypes, clinical isolates and also different Streptococcus species; second, we aimed to use microarray technology to examine the mechanisms by which environmental factors influence pneumococcal gene expression, and ultimately to further the understanding of how these changes in gene expression are achieved and how they may alter the virulence of the organism.

  20. Translation quality control is maintained by the penicillin resistance factor MurM in Streptococcus pneumoniae

    Shepherd, Jennifer; Ibba, Michael

    2013-01-01

    Streptococcus pneumoniae is a causative agent of nosocomial infections such as pneumonia, meningitis and septicaemia. Penicillin resistance in S. pneumoniae depends in part upon MurM, an aminoacyl-tRNA-ligase that attaches L-serine or L-alanine to the stem peptide lysine of Lipid II in cell wall...... combination of both branched and linear muropeptides, deletion of MurM results in a reversion to penicillin sensitivity in strains that were previously resistant. However, since MurM is not required for cell viability, the reason for its functional conservation across all strains of S. pneumoniae has remained...

  1. Enhanced detection and serotyping of Streptococcus pneumoniae using multiplex polymerase chain reaction

    Jong Gyun Ahn

    2012-11-01

    Full Text Available &lt;B&gt;Purpose:&lt;/B&gt; Methods for quick and reliable detection of &lt;I&gt;Streptococcus pneumoniae&lt;/I&gt; are needed for the diagnosis of pneumococcal disease and vaccine studies. This study aimed to show that sequential multiplex polymerase chain reaction (PCR is more efficient than conventional culture in achieving &lt;I&gt;S. pneumoniae -positive&lt;/i&gt; results. &lt;B&gt;Methods:&lt;/B&gt; Nasopharyngeal (NP secretions were obtained from 842 pediatric patients admitted with lower respiratory infections at Severance Children’s Hospital in Korea between March 2009 and June 2010. For identification and serotype determination of pneumococci from the NP secretions, the secretions were evaluated via multiplex PCR technique with 35 serotype-specific primers arranged in 8 multiplex PCR sets and conventional bacteriological culture technique. &lt;B&gt;Results:&lt;/B&gt; Among the results for 793 samples that underwent both bacterial culture and PCR analysis for pneumococcal detection, 153 (19.3% results obtained by PCR and 81 (10.2% results obtained by conventional culture technique were positive for S. pneumoniae. The predominant serotypes observed, in order of decreasing frequency, were 19A (23%, 6A/B (16%, 19F (11%, 15B/C (5%, 15A (5%, and 11A (4%; further, 26% of the isolates were non-typeable. &lt;B&gt;Conclusion:&lt;/B&gt; As opposed to conventional bacteriological tests, PCR analysis can accurately and rapidly identify pneumococcal serotypes.

  2. Lung abscess due to Streptococcus pneumoniae simulating pulmonary tuberculosis: presentation of two cases

    Alessandro Perazzo

    2014-03-01

    Full Text Available In the past, anaerobes were the most common cause of community-acquired lung abscess; Streptococcus species were the second most common cause. In recent years, this has changed. Klebsiella pneumoniae is now most common cause of community- acquired lung abscess, although Streptococcus species remain pathogen of major importance. We present two cases of pulmonary cavitation due to Streptococcus pneumoniae which resembled pulmonary tuberculosis with regards to their history and radiological findings. These are examples of a common diagnosis presenting in an uncommon way. Our cases had some peculiarities: they had a clinical picture strongly suggestive of pulmonary tuberculosis or lung cancer rather than necrotizing infectious pneumonia in patients with no comorbidities or underlying diseases (including oral or dental pathologies. Radiological findings did not help the clinicians: pulmonary tuberculosis was the first diagnostic hypothesis in both cases. An underlying lung cancer was excluded in the first case only after invasive pulmonary procedures.

  3. Serotype Distribution, Antimicrobial Susceptibility, and Molecular Epidemiology of Streptococcus pneumoniae Isolated from Children in Shanghai, China.

    Fen Pan

    Full Text Available Streptococcus pneumoniae is a common pathogenic cause of pediatric infections. This study investigated the serotype distribution, antimicrobial susceptibility, and molecular epidemiology of pneumococci before the introduction of conjugate vaccines in Shanghai, China.A total of 284 clinical pneumococcal isolates (270, 5, 4,3, and 2 of which were isolated from sputum, bronchoalveolar lavage fluid, blood, cerebral spinal fluid, and ear secretions, respectively from children less than 14 years of age who had not been vaccinated with a conjugate vaccine, were collected between January and December in 2013. All isolates were serotyped by multiplex polymerase chain reaction or quellung reactions and antimicrobial susceptibility testing was performed using the broth microdilution method. The molecular epidemiology of S.pneumoniae was analyzed by multilocus sequence typing (MLST.Among the 284 pneumococcal isolates, 19F (33.5%, 19A (14.1%, 23F (12.0%, and 6A (8.8% were the most common serotypes and the coverage rates of the 7-, 10-, and 13-valent pneumococcal conjugate vaccines (PCV7, PCV10, and PCV13 were 58.6%, 59.4% and 85.1%, respectively. Antimicrobial susceptibility showed that the prevalence rates of S.pneumoniae resistance to penicillin were 11.3% (32/284. Approximately 88.0% (250/284 of the isolates exhibited multi-drug resistance. MLST analysis revealed a high level of diversity, with 65 sequence types (STs among 267 isolates. Specifically, the four predominant STs were ST271 (24.3%, 65/267, ST320 (11.2%, 30/267, ST81 (9.7%, 26/267, and ST3173 (5.2%, 14/267, which were mainly associated with serotypes 19F, 19A, 23F, and 6A, respectively.The prevalent serotypes among clinical isolates from children were 19F, 19A, 23F, and 6A and these isolates showed high resistance rates to β-lactams and macrolides. The Taiwan19F-14 clone played a predominant role in the dissemination of pneumococcal isolates in Shanghai, China. Therefore, continued and

  4. Effect of Xylitol on Growth of Streptococcus pneumoniae in the Presence of Fructose and Sorbitol

    Tapiainen, Terhi; Kontiokari, Tero; Sammalkivi, Laura; Ikäheimo, Irma; Koskela, Markku; Uhari, Matti

    2001-01-01

    Xylitol is effective in preventing acute otitis media by inhibiting the growth of Streptococcus pneumoniae. To clarify this inhibition we used fructose, which is known to block similar growth inhibition observed in Streptococcus mutans. In addition, we evaluated the efficacy of sorbitol in inhibiting the growth of pneumococci, as sorbitol is widely used for indications similar to those for which xylitol is used. The addition of 5% xylitol to the growth medium resulted in marked growth inhibit...

  5. Streptococcus pneumoniae meningitis in Alberta pre- and postintroduction of the 7-valent pneumococcal conjugate vaccine

    Jennie Johnstone

    2011-01-01

    Full Text Available The objective of this study was to describe the epidemiology, clinical characteristics, microbiology and outcomes of patients of all ages with Streptococcus pneumoniae meningitis between 2000 and 2004; two years pre- and postintroduction of an S pneumoniae 7-valent conjugate vaccine program in Alberta in children younger than two years of age. The high mortality rate associated with S pneumoniae meningitis, despite appropriate therapy, suggests that prevention of S pneumoniae meningitis is critical. Despite implementation of a PCV-7 program in Alberta, rates of S pneumoniae meningitis in children younger than two years of age is still high. Thus, continued research into safe and efficacious vaccines covering a broader range of S pneumoniae serotypes is necessary.

  6. Comparative study of five different DNA fingerprint techniques for molecular typing of Streptococcus pneumoniae strains.

    Hermans, P W; Sluijter, M.; Hoogenboezem, T.; Heersma, H.; van Belkum, A; de Groot, R.

    1995-01-01

    The aim of this study was to identify the strengths and weaknesses of five DNA fingerprint methods for epidemiological typing of Streptococcus pneumoniae. We investigated the usefulness of (i) ribotyping, (ii) BOX fingerprinting with the BOX repetitive sequence of S. pneumoniae as a DNA probe, (iii) PCR fingerprinting with a primer homologous to the enterobacterial repetitive intergenic consensus sequence, (iv) pulsed-field gel electrophoresis of large DNA fragments, and (v) restriction fragm...

  7. Comparative study of five different DNA fingerprint techniques for molecular typing of Streptococcus pneumoniae strains

    Hermans, Peter; Sluijter, Marcel; Hoogenboezem, Theo; Heersma, H.; van Belkum, Alex; de Groot, Ronald

    1995-01-01

    textabstractThe aim of this study was to identify the strengths and weaknesses of five DNA fingerprint methods for epidemiological typing of Streptococcus pneumoniae. We investigated the usefulness of (i) ribotyping, (ii) BOX fingerprinting with the BOX repetitive sequence of S. pneumoniae as a DNA probe, (iii) PCR fingerprinting with a primer homologous to the enterobacterial repetitive intergenic consensus sequence, (iv) pulsed-field gel electrophoresis of large DNA fragments, and (v) restr...

  8. Reconstruction of a genome-scale metabolic network for Streptococcus pneumoniae R6

    J.P. Saraiva; Pinto, Francisco; Rocha, I

    2013-01-01

    The gram-positive, lancet-shaped bacteria Streptococcus pneumoniae thrives in almost any environment. Under certain conditions this pathogen can cause several infections such as meningitis, otitis media, endocarditis or pneumonia. Genome-scale metabolic networks (GSMs) are commonly used to study phenotype-genotype relationships using biochemical, physiological and genomic information. These relationships might shed some light on identification of targets for metabolic engineering or, in t...

  9. Ethanol Impairs Mucosal Immunity against Streptococcus pneumoniae Infection by Disrupting Interleukin 17 Gene Expression

    Trevejo-Nunez, Giraldina; Chen, Kong; Dufour, Jason P.; Bagby, Gregory J.; Horne, William T.; Nelson, Steve; Kolls, Jay K.

    2015-01-01

    Acute ethanol intoxication suppresses the host immune responses against Streptococcus pneumoniae. As interleukin 17 (IL-17) is a critical cytokine in host defense against extracellular pathogens, including S. pneumoniae, we hypothesized that ethanol impairs mucosal immunity against this pathogen by disrupting IL-17 production or IL-17 receptor (IL-17R) signaling. A chronic ethanol feeding model in simian immunodeficiency virus (SIV)-infected rhesus macaques and acute ethanol intoxication in a...

  10. Human Monocytes Promote Th1 and Th17 Responses to Streptococcus pneumoniae

    Olliver, Marie; Hiew, Jeffni; Mellroth, Peter; Henriques-Normark, Birgitta; Bergman, Peter

    2011-01-01

    Streptococcus pneumoniae is a leading cause of bacterial pneumonia, meningitis, and sepsis in children. Human immunity to pneumococcal infections has been assumed to depend on anticapsular antibodies. However, recent findings from murine models suggest that alternative mechanisms, dependent on T helper cells, are also involved. Although the immunological events in which T helper cells contribute to acquired immunity have been studied in mice, little is known about how these responses are gene...

  11. Multidrug-Resistant Streptococcus pneumoniae in Poland: Identification of Emerging Clones

    Overweg, Karin; Hermans, Peter W. M.; Trzcinski, Krzysztof; Sluijter, Marcel; De Groot, Ronald; Hryniewicz, Waleria

    1999-01-01

    Penicillin resistance among Streptococcus pneumoniae isolates has rapidly emerged in Poland during the last decade and has reached prevalence levels of up to 14.4% in 1997. In order to investigate the nature of this increase, a molecular epidemiological analysis of non-penicillin-susceptible multidrug-resistant pneumococci isolated in 1995 and 1996 was conducted. Thirty-seven patients who suffered mainly from upper respiratory tract infections and pneumococcal pneumonia were enrolled in this ...

  12. Immunization with Polyamine Transport Protein PotD Protects Mice against Systemic Infection with Streptococcus pneumoniae

    Shah, P.; Swiatlo, E.

    2006-01-01

    The human pathogen Streptococcus pneumoniae contains genes for a putative polyamine ABC transporter which are organized in an operon and designated potABCD. Polyamine transport protein D (PotD) is an extracellular protein which binds polyamines and possibly other structurally related molecules. PotD has been shown to contribute to virulence in both a murine sepsis model and a pneumonia model with capsular type 3 pneumococci. The protective efficacy of recombinant PotD was evaluated by active ...

  13. Multidrug-resistant Streptococcus pneumoniae in Poland: identification of emerging clones

    Overweg, Karin; Hermans, Peter; Trzcinski, K.; Sluijter, Marcel; Hryniewicz, W.

    1999-01-01

    textabstractPenicillin resistance among Streptococcus pneumoniae isolates has rapidly emerged in Poland during the last decade and has reached prevalence levels of up to 14.4% in 1997. In order to investigate the nature of this increase, a molecular epidemiological analysis of non-penicillin-susceptible multidrug-resistant pneumococci isolated in 1995 and 1996 was conducted. Thirty-seven patients who suffered mainly from upper respiratory tract infections and pneumococcal pneumonia were enrol...

  14. Exposure of Thomsen-Friedenreich Antigen in Streptococcus pneumoniae Infection is Dependent on Pneumococcal Neuraminidase A**

    Coats, Mamie T.; Murphy, Trudy; James C Paton; Gray, Barry; Briles, David E.

    2011-01-01

    Pneumococcal hemolytic uremic syndrome is recognized in a small portion of otherwise healthy children who have or have recently had Streptococcus pneumoniae infections, including severe pneumonia, meningitis, and bacteremia. As in other types of hemolytic uremic syndrome (HUS), pneumococcal HUS is characterized by microangiopathic hemolytic anemia, and thrombocytopenia, usually with extensive kidney damage. Although not demonstrated in vivo, the pathogenesis of pneumococcal HUS has been attri...

  15. Radiolabeling of gemifloxacin with technetium-99m and biological evaluation in artificially Streptococcus pneumoniae infected rats

    In the current investigation complexation of the gemifloxacin (GIN) with technetium-99 m (99mTc) and its biological evaluation in artificially Streptococcus pneumoniae (S. pneumoniae) infected rats was assessed as potential S. pneumoniae infection radiotracer. Radiochemically the 99mTc-GIN complex was further analyzed in terms of stability in saline, in vitro stability in serum at 37 deg C, in vitro binding with S. pneumoniae and biodistribution in artificially S. pneumoniae (living and heat killed) infected rats. The complex was found 97.25 ± 0.25% radiochemically stable in saline at 30 min after reconstitution. The stability of the 99mTc-GIN complex was decreased to 90.50 ± 0.20% within 240 min after reconstitution. In serum the 99mTc-GIN complex showed stable profile with the appearance of 18.85% free tracer within 16 h of incubation. The 99mTc-GIN complex showed saturated in vitro binding with S. pneumoniae after different intervals. Almost five fold uptake was observed in living S. pneumoniae infected muscle of the rats as compared to the inflamed and normal muscle. No significant difference in the uptake of heat killed S. pneumoniae infected, inflamed and normal muscles of the rats. The high RCP yield in saline, in vitro permanence in serum, in vitro binding with living S. pneumoniae and biodistribution in artificially S. pneumoniae infected rats we recommend the 99mTc-GIN as potential S. pneumoniae infection radiotracer. (author)

  16. Drug-resistance in Streptococcus pneumoniae isolates among Spanish middle aged and older adults with community-acquired pneumonia

    Raga-Luria Xavier

    2009-03-01

    Full Text Available Abstract Background Pneumococcal diseases remain a major cause of morbidity and mortality worldwide. Updated data on drug-resistance from different populations may be important to recognize changes in disease patterns. This study assessed current levels of penicilin resistance among Streptococcus Pneumoniae causing pneumonia in Spanish middle age and older adults. Methods Antimicrobial susceptibility was tested for 104 consecutive isolates of Streptococcus pneumoniae recovered from patients 50 years or older with radiographically confirmed pneumonia in the region of Tarragona (Spain between 2002 and 2007. According to the minimum inhibitory concentration of tested antimicrobials (penicillin, erythromycin, cefotaxime and levofloxacin strains were classified as susceptible or resistant. Antimicrobial resistance was determined for early cases (2002–2004 and contemporary cases (2005–2007. Results Twenty-seven (25.9% were penicillin-resistant strains (19 strains with intermediate resistance and 8 strains with high resistance. Penicillin-resistance was higher in 2002–2004 than in 2005–2007 (39.5% vs 18.2%, p = 0.017. Of 27 penicillin-resistant strains, 10 (37% were resistant to erythromycin, 8 (29.6% to cefotaxime, 2 (7.4% to levofloxacin, and 4 (14.8% were identified as multidrug resistant. Case-fatality rate was higher among those patients who had an infection caused by any penicillin susceptible strain (16.9% than in those with infections due to penicillin-resistant strains. Conclusion Resistance to penicillin among Streptococcus pneumoniae remains high, but such resistance does not result in increased mortality in patients with pneumococcal pneumonia.

  17. Resistance of Streptococcus pneumoniae to Deformylase Inhibitors Is Due to Mutations in defB

    Margolis, Peter; Hackbarth, Corinne; Lopez, Sara; Maniar, Mita; Wang, Wen; Yuan, Zhengyu; White, Richard; Trias, Joaquim

    2001-01-01

    Resistance to peptide deformylase inhibitors in Escherichia coli or Staphylococcus aureus is due to inactivation of transformylase activity. Knockout experiments in Streptococcus pneumoniae R6x indicate that the transformylase (fmt) and deformylase (defB) genes are essential and that a def paralog (defA) is not. Actinonin-resistant mutants of S. pneumoniae ATCC 49619 harbor mutations in defB but not in fmt. Reintroduction of the mutated defB gene into wild-type S. pneumoniae R6x recreates the...

  18. Agar diffusion tests with cefuroxime disks for predicting ceftriaxone susceptibility among isolates of Streptococcus pneumoniae

    Dias Cícero A.G.

    1998-01-01

    Full Text Available The performance of agar diffusion tests using disks of cefuroxime (30µg for predicting ceftriaxone susceptibility in 33 isolates of Streptococcus pneumoniae was studied. All 7 resistant isolates to ceftriaxone (MIC ³1.0 µg/ml exhibited zones of inhibition <28mm. The procedure can be easily adapted to clinical laboratories.

  19. Pulsatile Delivery of Clarithromycin Alone or in Combination with Amoxicillin against Streptococcus pneumoniae

    Leuthner, Kimberly D.; Cheung, Chrissy M.; Rybak, Michael J.

    2006-01-01

    We evaluated pulsatile dosing of clarithromycin and amoxicillin alone or combined against Streptococcus pneumoniae with various susceptibilities. When combined, pulsatile amoxicillin with clarithromycin was superior to either 8- or 12-h dosing against the intermediate strain and was identical for the susceptible strain. Pulse dosing of antimicrobials warrants further investigation.

  20. Selection of Streptococcus pneumoniae Mutants Having Reduced Susceptibility to Moxifloxacin and Levofloxacin

    Li, Xinying; Zhao, Xilin; Drlica, Karl

    2002-01-01

    With Streptococcus pneumoniae, moxifloxacin was 4- and 10-fold more effective than levofloxacin at restricting selection of resistant mutants and at killing resistant mutants, respectively. The selection frequency for first-step topoisomerase mutants was 1,000 times lower for moxifloxacin than for levofloxacin; this difference was lost when second-step mutants were selected.

  1. Bartholinitis caused by Streptococcus pneumoniae : Case report and review of literature

    Parvathi S

    2009-04-01

    Full Text Available Most of the Bartholin′s gland abscesses have been thought to be caused by colonizing micro-organisms of the perineal region. We encountered an interesting case of acute Bartholins abscess caused by Streptococcus pneumoniae in a primigravida. The abscess was incised and drained. The patient was treated with Cefuroxime. This case is presented for its rarity.

  2. Draft Genome Sequence of the Streptococcus pneumoniae Avery Strain A66

    Hahn, Christoph; Harrison, Ewan M.; Parkhill, Julian; Holmes, Mark A.; Paterson, Gavin K.

    2015-01-01

    We have used HiSeq 2000 technology to generate a draft genome sequence of Streptococcus pneumoniae strain A66. This is a common study strain used in investigations of pneumococcal bacterium-host interactions and was used in the seminal genetic studies of Avery et al.

  3. Pyruvate Oxidase Influences the Sugar Utilization Pattern and Capsule Production in Streptococcus pneumoniae

    Carvalho, Sandra M.; Farshchi Andisi, Vahid; Gradstedt, Henrik; Neef, Jolanda; Kuipers, Oscar P.; Neves, Ana R.; Bijlsma, Jetta J. E.

    2013-01-01

    Pyruvate oxidase is a key function in the metabolism and lifestyle of many lactic acid bacteria and its activity depends on the presence of environmental oxygen. In Streptococcus pneumoniae the protein has been suggested to play a major role in metabolism and has been implicated in virulence, oxidat

  4. CodY of Streptococcus pneumoniae : Link between nutritional gene regulation and colonization

    Hendriksen, Wouter T.; Bootsma, Hester J.; Estevao, Silvia; Hoogenboezem, Theo; de Jong, Anne; de Groot, Ronald; Kuipers, Oscar P.; Hermans, Peter W. M.

    2008-01-01

    CodY is a nutritional regulator mainly involved in amino acid metabolism. It has been extensively studied in Bacillus subtilis and Lactococcus lactis. We investigated the role of CodY in gene regulation and virulence of the human pathogen Streptococcus pneumoniae. We constructed a codY mutant and ex

  5. Carbonic Anhydrase Is Essential for Streptococcus pneumoniae Growth in Environmental Ambient Air

    Burghout, Peter; Cron, Lorelei E.; Gradstedt, Henrik; Quintero, Beatriz; Simonetti, Elles; Bijlsma, Jetta J. E.; Bootsma, Hester J.; Hermans, Peter W. M.

    2010-01-01

    The respiratory tract pathogen Streptococcus pneumoniae needs to adapt to the different levels of carbon dioxide (CO(2)) it encounters during transmission, colonization, and infection. Since CO(2) is important for various cellular processes, factors that allow optimal CO(2) sequestering are likely t

  6. To have neighbour's fare : extending the molecular toolbox for Streptococcus pneumoniae

    Kloosterman, TG; Bijlsma, J.J.E.; Kok, J; Kuipers, OP

    2006-01-01

    In past years, several useful genetic tools have been developed to study the molecular biology of Streptococcus pneumoniae. In order to extend the existing spectrum of tools, advantage was taken of the toolbox originally developed for the closely related bacterium Lactococcus lactis, which was adapt

  7. Antagonism between penicillin and erythromycin against Streptococcus pneumoniae in vitro and in vivo

    Johansen, H K; Jensen, T G; Dessau, Ram;

    2000-01-01

    effect of the bactericidal agent. In this study, the possible interaction between penicillin and erythromycin was investigated in vitro and in vivo against four clinical isolates of Streptococcus pneumoniae with MICs of penicillin ranging from 0.016 to 0.5 mg/L and of erythromycin from 0. 25 to >128 mg...

  8. Accuracy of Phenotypic Methods for Identification of Streptococcus pneumoniae Isolates Included in Surveillance Programs▿

    Richter, Sandra S.; Heilmann, Kristopher P.; Dohrn, Cassie L.; Riahi, Fathollah; Beekmann, Susan E.; Doern, Gary V.

    2008-01-01

    Similarities between Streptococcus pneumoniae and viridans group streptococci may result in misidentification of these organisms. In surveillance programs which assess antimicrobial resistance rates among respiratory tract pathogens, such identification errors could lead to overestimates of pneumococcal resistance rates. DNA probe analysis (Gen-Probe, San Diego, CA), the bile solubility test, optochin susceptibility, colony morphology, and the capsular swelling reaction with Omni serum (State...

  9. Multidrug-resistant Streptococcus pneumoniae isolates from healthy Ghanaian preschool children

    Dayie, Nicholas Tete Kwaku Dzifa; Arhin, Reuben E.; Newman, Mercy J.;

    2015-01-01

    Streptococcus pneumoniae is the cause of high mortality among children worldwide. Antimicrobial treatment and vaccination are used to control pneumococcal infections. In Ghana, data on antimicrobial resistance and the prevalence of multidrug-resistant pneumococcal clones are scarce; hence, the ai...

  10. Accuracy of using the lytA gene to distinguish Streptococcus pneumoniae from related species

    Greve, Thomas; Møller, Jens Kjølseth

    2012-01-01

    The need for a microbial identification of Streptococcus pneumoniae independent of culture methods has resulted in the introduction of other laboratory principles. The verification of a proper and exclusive gene for the detection of the pneumococcus by the nucleic acid-based tests (NAT) is however...

  11. Structure and molecular characterization of Streptococcus pneumoniae capsular polysaccharide 10F by carbohydrate engineering in Streptococcus oralis.

    Yang, Jinghua; Shelat, Nirav Y; Bush, C Allen; Cisar, John O

    2010-07-30

    Although closely related at the molecular level, the capsular polysaccharide (CPS) of serotype 10F Streptococcus pneumoniae and coaggregation receptor polysaccharide (RPS) of Streptococcus oralis C104 have distinct ecological roles. CPS prevents phagocytosis of pathogenic S. pneumoniae, whereas RPS of commensal S. oralis functions as a receptor for lectin-like adhesins on other members of the dental plaque biofilm community. Results from high resolution NMR identified the recognition region of S. oralis RPS (i.e. Galfbeta1-6GalNAcbeta1-3Galalpha) in the hexasaccharide repeat of S. pneumoniae CPS10F. The failure of this polysaccharide to support fimbriae-mediated adhesion of Actinomyces naeslundii was explained by the position of Galf, which occurred as a branch in CPS10F rather than within the linear polysaccharide chain, as in RPS. Carbohydrate engineering of S. oralis RPS with wzy from S. pneumoniae attributed formation of the Galf branch in CPS10F to the linkage of adjacent repeating units through sub terminal GalNAc in Galfbeta1-6GalNAcbeta1-3Galalpha rather than through terminal Galf, as in RPS. A gene (wcrD) from serotype 10A S. pneumoniae was then used to engineer a linear surface polysaccharide in S. oralis that was identical to RPS except for the presence of a beta1-3 linkage between Galf and GalNAcbeta1-3Galalpha. This polysaccharide also failed to support adhesion of A. naeslundii, thereby establishing the essential role of beta1-6-linked Galf in recognition of adjacent GalNAcbeta1-3Galalpha in wild-type RPS. These findings, which illustrate a molecular approach for relating bacterial polysaccharide structure to function, provide insight into the possible evolution of S. oralis RPS from S. pneumoniae CPS. PMID:20507989

  12. Antimicrobial resistance and serotyping of Streptococcus pneumoniae isolated from pediatric patients in Belo Horizonte, MG, Brazil Resistência antimicrobiana e sorotipagem de Streptococcus pneumoniae isolado de pacientes pediátricos em Belo Horizonte, MG

    Ana Paula Gomes de Oliveira Magalhães

    2003-07-01

    Full Text Available Thirty one Streptococcus pneumoniae invasive strains were isolated from a pediatric population in Belo Horizonte from June, 1999 to May, 2001. Penicillin, trimethoprim-sulfamethoxazole, tetracycline and chloramphenicol resistance rates for the isolates were 41.9, 58.1, 25.8 and 3.2%, respectively. Intermediate penicillin resistant (MICs between 0.1 and 1.0 µg/ml and resistant (MICs > 2.0 µg/ml isolates occured at rates of 38.7 and 3.2%, respectively. Resistance to erythromycin, ofloxacin, rifampin or vancomicyn was not detected. Ten S. pneumoniae serotypes (14, 5, 10 A, 6B, 15B, 18C, 6 A, 18 A, 19 A and 19 F were identified. Serotype 14 (12 out of 31 was predominant among the isolates. Penicillin and trimethoprim-sulfamethoxazole resistance was more common in 14 and 6B serotypes.Trinta e três linhagens invasivas do S. pneumoniae foram isoladas a partir de pacientes pediátricos em Belo Horizonte, MG, Brasil, de junho de 1999 a maio de 2001. As taxas de resistência à penicilina, ao trimetoprim-sultametoxazol, tetraciclina e cloranfenicol foram respectivamente, 41, 9; 58,1 e 3,2%. A resistência intermediária à penicilina (MICs entre 0,1 e 1,0 µg/ml e resistência total (MICs>2.0 µg/ml ocorreram, respectivamente, nas porcentagens de 38,7 e 3,2%. Não foi detectada resistência à eritromicina, ofloxacin, rifampina e vancomicina. Foram identificados 9 sorotipos do S. pneumoniae (14, 5, 10 , 6B, 15B, 18C, 6 A, 18 19 A e 19F entre os isolados. O sorotipo 14 (12 de 31 foi predominate entre os isolados. A resistência à penicilina e ao trimetoprim-sulfametoxazol estava sempre associada aos sorotipos 14 e 6B.

  13. EARSS: European Antimicrobial Resistance Surveillance System; data from the Netherlands .Incidence and resistance rates for Streptococcus pneumoniae and Staphylococcus aureus

    Goettsch WG; de Neeling AJ; CIE; LIO

    2001-01-01

    Gevoeligheid voor antimicrobiele middelen in Streptococcus pneumoniae en Staphylococcus aureus werd bepaald in 1999 in Nederland binnen het raamwerk van het European antomicrobial Resistance Surveillance System (EARSS). Het EARSS project had in Nederland een dekkingsgraad van 40% van de Nederlandse

  14. Th1-directing adjuvants increase the immunogenicity of oligosaccharide-protein conjugate vaccines related to Streptococcus pneumoniae type 3

    Lefeber, DJ; Benaissa-Trouw, B; Vliegenthart, JFG; Kamerling, JP; Jansen, WTM; Kraaijeveld, K; Snippe, H

    2003-01-01

    Oligosaccharide (OS)-protein conjugates are promising candidate vaccines against encapsulated bacteria, such as Haemophilus influenzae, Neisseria meningitidis, and Streptococcus pneumoniae. Although the effects of several variables such as OS chain length and protein carrier have been studied, littl

  15. Evaluation of Streptococcus pneumoniae in bile samples: A case series review.

    Itoh, Naoya; Kawamura, Ichiro; Tsukahara, Mika; Mori, Keita; Kurai, Hanako

    2016-06-01

    Although Streptococcus pneumoniae is an important pathogen of humans, pneumococcal cholangitis is rare because of the rapid autolysis of S. pneumoniae. The aim of this case series was to review patients with bile cultures positive for S. pneumoniae. This study was a single center retrospective case series review of patients with S. pneumoniae in their bile at a tertiary-care cancer center between September 2002 and August 2015. Subjects consisted of all patients in whom S. pneumoniae was isolated in their bile during the study period. Bile specimens for culture were obtained from biliary drainage procedures such as endoscopic retrograde biliary drainage, endoscopic nasobiliary drainage, and percutaneous transhepatic biliary drainage. There were 20 patients with bile cultures positive for S. pneumoniae during the study period. All patients presented with extrahepatic obstructive jaundice due to hepatopancreatobiliary tumors. Nineteen of 20 patients underwent the placement of plastic intrabiliary tubes. The mean time between the first-time drainage and the positive culture was 26 days (range 0-313 days). Although 12 of 20 patients met our definition of cholangitis, 5 were clinically treated with antibiotics based on a physician's assessment of whether there was a true infection. The present study is the largest case series of patients with S. pneumoniae in their bile. Based on our findings, the isolation of S. pneumoniae from bile may be attributed to the placement of biliary drainage devices. PMID:27025902

  16. SURFACE PROTEINS AND PNEUMOLYSIN OF ENCAPSULATED AND NONENCAPSULATED STREPTOCOCCUS PNEUMONIAE MEDIATE VIRULENCE IN A CHINCHILLA MODEL OF OTITIS MEDIA

    Keller, Lance E.; Bradshaw, Jessica L.; Haley ePipkins; McDaniel, Larry S.

    2016-01-01

    Streptococcus pneumoniae infections result in a range of human diseases and are responsible for almost one million deaths annually. Pneumococcal disease is mediated in part through surface structures and an anti-phagocytic capsule. Recent studies have shown that nonencapsulated Streptococcus pneumoniae (NESp) make up a significant portion of the pneumococcal population and are able to cause disease. NESp lack some common surface proteins expressed by encapsulated pneumococci, but express surf...

  17. Pelvic inflammatory disease due to Streptococcus pneumoniae: a usual pathogen at an unusual place.

    Lemoyne, S; Van Leemput, J; Smet, D; Desmedt, E; Devos, H; Van Schaeren, J; Jeurissen, A

    2008-01-01

    We report three cases of pelvic inflammatory disease (PID) due to Streptococcus pneumoniae in previously healthy young women. S. pneumoniae frequently causes bacteremia, meningitis and respiratory infections, but it very rarely infects the genital tract. All our patients presented with an acute onset of severe abdominal pain and had an intrauterine device (IUD) present. No abnormal sexual behavior was noticed. Although the relation between PID due to S. pneumoniae and the use of an IUD has been a topic for discussions, culture of IUD in all our patients and blood culture in 2 of 3 of our patients revealed S. pneumoniae. All patients recovered well with intravenous antibiotic treatment and removal of the IUD. PMID:19170357

  18. Search for coherent gene modules that predict streptococcus pneumoniae strain invasiveness

    Catarino, Rui Ribeiro

    2012-01-01

    Tese de mestrado em Bioquímica, apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2012 O Streptococcus pneumoniae, também chamado pneumococcus, é uma bactéria grampositiva do subgrupo alfa-hemolítico do género Streptococcus. É um colonizador frequente do trato respiratório superior humano e embora possa ser encontrado em qualquer pessoa, tem maior prevalência em crianças e idosos. A colonização decorre tipicamente sem causar sintomas, mas pode por vezes culminar na in...

  19. Immune Responses to Specific Antigens of Streptococcus pneumoniae and Moraxella catarrhalis in the Respiratory Tract

    Samukawa, Takao; Yamanaka, Noboru; Hollingshead, Susan; Klingman, Karin; Faden, Howard

    2000-01-01

    Streptococcus pneumoniae and Moraxella catarrhalis are two common respiratory pathogens, colonizing as many as 54 and 72% of children, respectively, by 1 year of age. The immune responses to surface protein A of S. pneumoniae (PspA) and the high-molecular-weight outer membrane protein of M. catarrhalis (UspA) in the sera of various age groups in the general population and in the nasopharynges of 30 children monitored from birth through 1 year of age were evaluated. Immunoglobulin G (IgG) was ...

  20. Laboratory survey of drug-resistant Streptococcus pneumoniae in New York City, 1993-1995.

    Heffernan, R.; Henning, K; Labowitz, A.; Hjelte, A.; Layton, M.

    1998-01-01

    Wide geographic variation in the prevalence of drug-resistant Streptococcus pneumoniae demonstrates the importance of tracking antimicrobial resistance locally. This survey of hospital microbiology laboratories in New York City found that penicillin resistance (MIC > or = 2.0 micrograms/ml) increased from 1.5% of S. pneumoniae isolates in 1993 to 6.3% in 1995 and that in 1995, one-third of isolates nonsusceptible to penicillin (MIC > or = 0.1 microgram/ml) were also nonsusceptible to an exten...

  1. Prevalência de sorotipos e resistência antimicrobiana de cepas invasivas do Streptococcus pneumoniae

    Mantese Orlando C.

    2003-01-01

    Full Text Available OBJETIVO: Avaliar o perfil de sorotipos e a susceptibilidade aos antimicrobianos de cepas de Streptococcus pneumoniae obtidas em espécimes clínicos de pacientes com doença invasiva, bem como suas implicações na formulação de vacinas pneumocócicas. MÉTODOS: Cepas de pneumococo isoladas no Laboratório de Análises Clínicas do Hospital de Clínicas da Universidade Federal de Uberlândia a partir de amostras clínicas de pacientes com doença invasiva foram identificadas e enviadas ao Instituto Adolfo Lutz em São Paulo para confirmação da identificação, sorotipagem e determinação da susceptibilidade aos antimicrobianos. RESULTADOS: De abril de 1999 a março de 2003, foram isoladas 148 cepas invasivas de pneumococo, sendo 84 (56,7% provenientes de pacientes do sexo masculino. A idade variou de um dia a 88,83 anos, com média de 21,33+25,82 anos e mediana de 4,42 anos. Os diagnósticos clínicos mais comuns foram pneumonia (91 casos; 61,4%, meningite (32 casos; 21,6% e bacteremia sem foco evidente (15 casos; 10,1%. As principais fontes de recuperação foram sangue (76 amostras; 51,3%, líquido pleural (39; 26,3% e liquor (30; 20,2%. No total, foram identificados 23 diferentes sorotipos entre 143 amostras testadas, sendo os mais comuns os seguintes: 14, 3, 1, 5, 6A, 6B e 18C. Dentre 30 (20,2% cepas oxacilina-resistentes, 23 (15,5% confirmaram a resistência à penicilina (12,8% com nível intermediário e 2,7%, com nível pleno, que esteve restrita aos sorotipos 14, 23F, 19A e 6B, predominando em indivíduos com até dois anos de idade (p = 0,0008. Foi detectada susceptibilidade diminuída ao cotrimoxazol (63,4%, à eritromicina (8,3%, à clindamicina (8,7% e à ofloxacina (0,8%. A resistência à cefotaxima foi detectada em três das 30 cepas testadas (2% das 148, todas elas com resistência confirmada à penicilina. Não foi observada resistência a cloranfenicol, rifampicina ou vancomicina. CONCLUSÕES: A resistência

  2. Disease Isolates of Streptococcus pseudopneumoniae and Non-Typeable S. pneumoniae Presumptively Identified as Atypical S. pneumoniae in Spain

    Dora Rolo; Simões, Alexandra S.; Arnau Domenech; Asunción Fenoll; Josefina Liñares; Hermínia de Lencastre; Carmen Ardanuy; Raquel Sá-Leão

    2013-01-01

    We aimed to obtain insights on the nature of a collection of isolates presumptively identified as atypical Streptococcus pneumoniae recovered from invasive and non-invasive infections in Spain. One-hundred and thirty-two isolates were characterized by: optochin susceptibility in ambient and CO(2)-enriched atmosphere; bile solubility; PCR-based assays targeting pneumococcal genes lytA, ply, pspA, cpsA, Spn9802, aliB-like ORF2, and a specific 16S rRNA region; multilocus sequence analysis; and a...

  3. Characterization of a mutation in the parE gene that confers fluoroquinolone resistance in Streptococcus pneumoniae.

    Perichon, B; Tankovic, J; Courvalin, P

    1997-01-01

    We report a mutation in the parE genes of two in vitro mutants of Streptococcus pneumoniae responsible for low-level resistance to fluoroquinolones. Sequential acquisition of mutations in parE and gyrA leads to higher levels of resistance. This confirms that topoisomerase IV is the primary target of fluoroquinolones in S. pneumoniae.

  4. The pavA gene of Streptococcus pneumoniae encodes a fibronectin-binding protein that is essential for virulence

    Holmes, AR; McNab, R; Millsap, KW; Rohde, M; Hammerschmidt, S; Mawdsley, JL; Jenkinson, HF

    2001-01-01

    Streptococcus pneumoniae colonizes the nasopharynx in up to 40% of healthy subjects, and is a leading cause of middle ear infections (otitis media), meningitis and pneumonia. Pneumococci adhere to glycosidic receptors on epithelial cells and to immobilized fibronectin, but the bacterial adhesins med

  5. Streptococcus pneumoniae Invades Endothelial Host Cells via Multiple Pathways and Is Killed in a Lysosome Dependent Manner

    Gradstedt, Henrik; Iovino, Federico; Bijlsma, Jetta J. E.

    2013-01-01

    Streptococcus pneumoniae is one of the major causative agents of pneumonia, sepsis, meningitis and other morbidities. In spite of its heavy disease burden, surprisingly little is known about the mechanisms involved in the switch of life style, from commensal colonizer of the nasopharynx to invasive

  6. New Alkaloid Antibiotics That Target the DNA Topoisomerase I of Streptococcus pneumoniae

    Garcia, M. T.; Blazquez, M. A.; Ferrandiz, M. J.; Sanz, M. J.; Silva-Martin, N.; Hermoso, J. A.; De La Campa, A G

    2010-01-01

    Streptococcus pneumoniae has two type II DNA-topoisomerases (DNA-gyrase and DNA topoisomerase IV) and a single type I enzyme (DNA-topoisomerase I, TopA), as demonstrated here. Although fluoroquinolones target type II enzymes, antibiotics efficiently targeting TopA have not yet been reported. Eighteen alkaloids (seven aporphine and 11 phenanthrenes) were semisynthesized from boldine and used to test inhibition both of TopA activity and of cell growth. Two phenanthrenes (seconeolitsine and N-me...

  7. Activities against Streptococcus pneumoniae of amoxicillin and cefotaxime at physiological concentrations: in vitro pharmacodynamic simulation.

    Balcabao, I P; Aguilar, L.; Martín, M; García, Y.; Dal-Ré, R; Prieto, J.

    1996-01-01

    An in vitro model simulating amoxicillin and cefotaxime concentrations in human serum (after standard doses) was used to explore the activities of these drugs over time against penicillin-susceptible and penicillin-resistant Streptococcus pneumoniae strains. An initial inoculum reduction percentage of > or = 90% was obtained with amoxicillin and maintained for 2 to 8 h, regardless of the strain tested. In contrast, experiments showed that cefotaxime had significantly (P < 0.001) less capabili...

  8. Within-Host Selection Is Limited by an Effective Population of Streptococcus pneumoniae during Nasopharyngeal Colonization

    Li, Yuan; Thompson, Claudette M; Trzciński, Krzysztof; Lipsitch, Marc

    2013-01-01

    Streptococcus pneumoniae (pneumococcus) is a significant pathogen that frequently colonizes the human nasopharynx. Environmental factors, including antimicrobial use and host immunity, exert selection on members of the nasopharyngeal population, and the dynamics of selection are influenced by the effective population size of the selected population, about which little is known. We measured here the variance effective population size (N(e)) of pneumococcus in a mouse colonization model by moni...

  9. Comparative analysis of Streptococcus pneumoniae transmission in Portuguese and Finnish day-care centres

    Pessoa, Delphine; Hoti, Fabian; Syrjänen, Ritva; Sá-Leão, Raquel; Kaijalainen, Tarja; Gomes, M. Gabriela M.; Auranen, Kari

    2013-01-01

    Day-care centre (DCC) attendees play a central role in maintaining the circulation of Streptococcus pneumoniae (pneumococcus) in the population. The prevalence of pneumococcal carriage is highest in DCC attendees but varies across countries and is found to be consistently lower in Finland than in Portugal. We compared key parameters underlying pneumococcal transmission in DCCs to understand which of these contributed to the observed differences in carriage prevalence. PneumoCarr Consortium...

  10. Draft Genome Sequence of an Atypical Strain of Streptococcus pneumoniae Isolated from a Respiratory Infection

    Choi, Sang Chul; Parker, Jayme; Richards, Vincent P; Ross, Katherine; Jilly, Bernard; Chen, Jack

    2014-01-01

    Next-generation sequencing was used to investigate an unknown clinical respiratory infection. This new strain of Streptococcus pneumoniae, ASVL_JC_0001, was isolated from a clinical specimen from a patient with bronchitis and pulmonary inflammation. The draft genome sequence, obtained with an Illumina MiSeq sequencing system, consists of 83 large contigs, a total of 2,092,532 bp long, and has a GC content of 40.3%.

  11. Time to positivity in blood cultures of adults with Streptococcus pneumoniae bacteremia

    Ansorena Luis

    2006-04-01

    Full Text Available Abstract Background previous studies have established that bacterial blood concentration is related with clinical outcome. Time to positivity of blood cultures (TTP has relationship with bacterial blood concentration and could be related with prognosis. As there is scarce information about the usefulness of TTP, we study the relationship of TTP with clinical parameters in patients with Streptococcus pneumoniae bacteremia. Methods TTP of all cases of Streptococcus pneumoniae bacteremia, detected between January 1995 and December 2004 using the BacT/Alert automated blood culture system in a teaching community hospital was analyzed. When multiple cultures were positive only the shortest TTP was selected for the analysis. Results in the study period 105 patients with Streptococcus pneumoniae bacteremia were detected. Median TTP was 14.1 hours (range 1.2 h to 127 h. Immunosuppressed patients (n = 5, patients with confusion (n = 19, severe sepsis or shock at the time of blood culture extraction (n = 12, those with a diagnosis of meningitis (n = 7 and those admitted to the ICU (n = 14 had lower TTP. Patients with TTP in the first quartile were more frequently hospitalized, admitted to the ICU, had meningitis, a non-pneumonic origin of the bacteremia, and a higher number of positive blood cultures than patients with TTP in the fourth quartile. None of the patients with TTP in the 90th decile had any of these factors associated with shorter TTP, and eight out of ten patients with TTP in the 10th decile had at least one of these factors. The number of positive blood cultures had an inverse correlation with TTP, suggesting a relationship of TTP with bacterial blood concentration. Conclusion Our data support the relationship of TTP with several clinical parameters in patients with Streptococcus pneumoniae bacteremia, and its potential usefulness as a surrogate marker of outcome.

  12. The Protective Function of Human C-reactive Protein in Mouse Models of Streptococcus pneumoniae Infection

    Agrawal, Alok; Suresh, Madathilparambil V.; Singh, Sanjay K.; Ferguson, Donald A.

    2008-01-01

    Human C-reactive protein (CRP), injected intravenously into mice or produced inside mice by a human transgene, protects mice from death following administration of lethal numbers of Streptococcus pneumoniae. The protective effect of CRP is due to reduction in the concentration of bacteria in the blood. The exact mechanism of CRP-dependent killing of pneumococci and the partners of CRP in this process are yet to be defined. The current efforts to determine the mechanism of action of CRP in mic...

  13. Molecular Characterization and Antimicrobial Susceptibility of Fluoroquinolone-Resistant or -Susceptible Streptococcus pneumoniae from Hong Kong

    Morrissey, Ian; Farrell, David J.; Bakker, Sarah; Buckridge, Sylvie; Felmingham, David

    2003-01-01

    Fluoroquinolone resistance in Streptococcus pneumoniae isolated from Hong Kong as part of Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin 1999/2000 was found to be due to the spread of the Spain23F-1 clone (mainly a Spain23F-1-14 variant). All the isolates were multidrug resistant but were susceptible to quinupristin-dalfopristin, linezolid, and telithromycin. The Spain23F-1 clone also occurred among antimicrobial-susceptible isolates, which suggests th...

  14. Capsular-type prediction by phylogenetic tree of glycosyltransferase gene polymorphism in Streptococcus pneumoniae

    Tomita, Yuka

    2011-01-01

    Yuka Tomita1,2, Akira Okamoto2, Keiko Yamada2, Testuya Yagi3, Yoshinori Hasegawa4, Michio Ohta21Department of Infectious Disease, Nagoya University Hospital, 2Department of Bacteriology, Nagoya University Graduate School of Medicine, 3Center of National University Hospital for Infection Control, Nagoya University Hospital, 4Department of Respiratory Medicine, Nagoya University Graduate School of Medicine, Nagoya, JapanAbstract: Streptococcus pneumoniae can cause severe infections among childr...

  15. Mutational Analysis of the Streptococcus pneumoniae Bimodular Class A Penicillin-Binding Proteins

    Paik, Johanna; Kern, Iza; Lurz, Rudi; Hakenbeck, Regine

    1999-01-01

    One group of penicillin target enzymes, the class A high-molecular-weight penicillin-binding proteins (PBPs), are bimodular enzymes. In addition to a central penicillin-binding–transpeptidase domain, they contain an N-terminal putative glycosyltransferase domain. Mutations in the genes for each of the three Streptococcus pneumoniae class A PBPs, PBP1a, PBP1b, and PBP2a, were isolated by insertion duplication mutagenesis within the glycosyltransferase domain, documentin...

  16. Clustering of serotypes in a longitudinal study of Streptococcus pneumoniae carriage in three day care centres

    Tanskanen Antti; Syrjänen Ritva; Hoti Fabian; Leino Tuija; Auranen Kari

    2008-01-01

    Abstract Background Streptococcus pneumoniae (pneumococcus) causes a wide range of clinical manifestations that together constitute a major burden of disease worldwide. The main route of pneumococcal transmission is through asymptomatic colonisation of the nasopharynx. Studies of transmission are currently of general interest because of the impact of the new conjugate-polysaccharide vaccines on nasopharyngeal colonisation (carriage). Here we report the first longitudinal study of pneumococcal...

  17. Comparative analysis of Streptococcus pneumoniae transmission in Portuguese and Finnish day-care centres

    Pessoa, Delphine; Hoti, Fabian; Syrjänen, Ritva; Sá-Leão, Raquel; Kaijalainen, Tarja; Gomes, M. Gabriela M.; Auranen, Kari

    2013-01-01

    Background Day-care centre (DCC) attendees play a central role in maintaining the circulation of Streptococcus pneumoniae (pneumococcus) in the population. The prevalence of pneumococcal carriage is highest in DCC attendees but varies across countries and is found to be consistently lower in Finland than in Portugal. We compared key parameters underlying pneumococcal transmission in DCCs to understand which of these contributed to the observed differences in carriage prevalence. Methods Longi...

  18. Genetische Determinanten für die Resistenz von Streptococcus pneumoniae gegen das neue Antibiotikum Vancoresmycin

    Petra, Becker

    2008-01-01

    Bei dem in dieser Arbeit untersuchten Antibiotikum Vancoresmycin handelt es sich um ein neuartiges Tetramsäurederivat, das aus der Fermentationsbrühe des Aktinomyceten Amycolatopsis vancoresmycina gewonnen wurde. Da der Wirkungsmechanismus und mögliche Resistenzmechanismen unbekannt sind, soll diese Arbeit zu deren Aufklärung beitragen. Dazu wurden verschiedene Determinanten für die Resistenz gegen Vancoresmycin in Streptococcus pneumoniae R6 analysiert. Zuerst wurden acht vancoresmycinresist...

  19. Consumption Patterns and In Vitro Resistance of Streptococcus pneumoniae to Fluoroquinolones▿

    Simoens, Steven; Verhaegen, Jan; van Bleyenbergh, Pascal; Peetermans, Willy E; Decramer, Marc

    2011-01-01

    This article analyzes patterns of consumption of fluoroquinolones and documents the in vitro resistances of Streptococcus pneumoniae isolates to fluoroquinolones in the ambulatory care setting in Belgium over time. The volume of fluoroquinolone consumption has fallen consistently since 2003. Fluoroquinolones were used primarily for their registered indications (i.e., urinary tract infections and lower respiratory tract infections). The MIC distributions of moxifloxacin and levofloxacin in S. ...

  20. Oscillations in continuous culture populations of Streptococcus pneumoniae: population dynamics and the evolution of clonal suicide

    2008-01-01

    Agents that kill or induce suicide in the organisms that produce them or other individuals of the same genotype are intriguing puzzles for ecologists and evolutionary biologists. When those organisms are pathogenic bacteria, these suicidal toxins have the added appeal as candidates for the development of narrow spectrum antibiotics to kill the pathogens that produce them. We show that when clinical as well as laboratory strains of Streptococcus pneumoniae are maintained in continuous culture ...

  1. Variation in Streptococcus pneumoniae susceptibility to human antimicrobial peptides may mediate intraspecific competition

    Habets, Michelle G. J. L.; Rozen, Daniel E; Brockhurst, Michael A

    2012-01-01

    Streptococcus pneumoniae is a facultative pathogen inhabiting the nasopharynx of humans where it is exposed to a range of antimicrobial peptides (AMPs) of the innate immune response. It is possible therefore that the susceptibility of strains to AMPs plays a role in determining their ability to colonize, and furthermore, that AMPs could mediate competitive interactions between co-colonizing genotypes. However, little is known about patterns of natural variation in AMP susceptibility of S. pne...

  2. Regulation of Naturally Acquired Mucosal Immunity to Streptococcus pneumoniae in Healthy Malawian Adults and Children

    Sarah J Glennie; Banda, Dominic; Mulwafu, Wakisa; Nkhata, Rose; Neil A Williams; Heyderman, Robert S.

    2012-01-01

    Worldwide, invasive pneumococcal disease caused by Streptococcus pneumoniae is most common in young children. In adults, disease rates decline following intermittent colonization and the acquisition of naturally acquired immunity. We characterized mucosal and systemic pneumococcal-specific T-cell responses in African children and adults who contend with intense rates of colonization, up to 100% and 60% respectively. We find most Malawian children have high pneumococcal-specific T-cell respons...

  3. Identification of Pneumococcal Surface Protein A as a Lactoferrin-Binding Protein of Streptococcus pneumoniae

    Hammerschmidt, Sven; Bethe, Gesina; H. Remane, Petra; Chhatwal, Gursharan S.

    1999-01-01

    Lactoferrin (Lf), an iron-sequestering glycoprotein, predominates in mucosal secretions, where the level of free extracellular iron (10−18 M) is not sufficient for bacterial growth. This represents a mechanism of resistance to bacterial infections by prevention of colonization of the host by pathogens. In this study we were able to show that Streptococcus pneumoniae specifically recognizes and binds the iron carrier protein human Lf (hLf). Pretreatment of pneumococci with proteases reduced hL...

  4. The Capsule Sensitizes Streptococcus pneumoniae to α-Defensins Human Neutrophil Proteins 1 to 3▿

    Beiter, Katharina; Wartha, Florian; Hurwitz, Robert; Normark, Staffan; Zychlinsky, Arturo; Henriques-Normark, Birgitta

    2008-01-01

    Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide. Its polysaccharide capsule causes resistance to phagocytosis and interferes with the innate immune system's ability to clear infections at an early stage. Nevertheless, we found that encapsulated pneumococci are sensitive to killing by a human neutrophil granule extract. We fractionated the extract by high-performance liquid chromatography and identified α-defensins by mass spectrometry as the proteins responsible...

  5. Nasopharyngeal carriage of community-acquired, antibiotic-resistant Streptococcus pneumoniae in a Zambian paediatric population.

    Woolfson, A; Huebner, R.; Wasas, A; Chola, S.; Godfrey-Faussett, P.; Klugman, K.

    1997-01-01

    The emergence of antibiotic-resistant Streptococcus pneumoniae is an international health problem. Apart from South Africa few data on pneumococcal resistance are available for sub-Saharan Africa. This study examines the nasopharyngeal carriage and prevalence of antibiotic resistance in pneumococci isolated from 260 Zambian children aged < 6 years. Pneumococci were isolated from 71.9% of the children; the odds of carrying organisms were twice as high among children < 2 years of age compared w...

  6. Proteomic Study of Peptide Deformylase Inhibition in Streptococcus pneumoniae and Staphylococcus aureus

    Wang, Wen; White, Richard; Yuan, Zhengyu

    2006-01-01

    Peptide deformylase (PDF) is an essential enzyme in both gram-negative and gram-positive bacteria. It hydrolyzes formylated N-terminal peptides to generate free N-terminal peptides during the process of protein maturation. Inhibition of this enzyme results in cessation of bacterial growth. We have examined the effect of a potent PDF inhibitor, LBM-415 (also known as VIC-104959), on the proteomes of Staphylococcus aureus and Streptococcus pneumoniae using two-dimensional electrophoresis. Both ...

  7. Rapid identification of Staphylococcus aureus and Streptococcus pneumoniae from blood cultures.

    Knight, R G; Shlaes, D M

    1983-01-01

    Simultaneous application of the lysostaphin sensitivity test for identification of Staphylococcus aureus and the deoxycholate test for the identification of Streptococcus pneumoniae was evaluated for reliability in rapid identification (1 h) of these organisms from blood cultures by using BACTEC 6B and 7C bottles. The procedure was applied to 127 cultures, 74 lysostaphin tests and 53 deoxycholate tests. Lysostaphin-tested organisms included 23 S. aureus, 40 Staphylococcus epidermidis, 1 Liste...

  8. Pronounced Metabolic Changes in Adaptation to Biofilm Growth by Streptococcus pneumoniae

    Allan , RN; Skipp, Paul; Jefferies, J.M.C.; Clarke, S. C.; Faust, SN; Hall-Stoodley, L.

    2014-01-01

    Streptococcus pneumoniae accounts for a significant global burden of morbidity and mortality and biofilm development is increasingly recognised as important for colonization and infection. Analysis of protein expression patterns during biofilm development may therefore provide valuable insights to the understanding of pneumococcal persistence strategies and to improve vaccines. iTRAQ (isobaric tagging for relative and absolute quantification), a high-throughput gel-free proteomic approach whi...

  9. Validation of an Immunodiagnostic Assay for Detection of 13 Streptococcus pneumoniae Serotype-Specific Polysaccharides in Human Urine

    Pride, Michael W.; Huijts, Susanne M.; Wu, Kangjian; Souza, Victor; Passador, Sherry; Tinder, Chunyan; Song, Esther; Elfassy, Arik; McNeil, Lisa; Menton, Ronald; French, Roger; Callahan, Janice; Webber, Chris; Gruber, William C.; Bonten, Marc J. M.

    2012-01-01

    To improve the clinical diagnosis of pneumococcal infection in bacteremic and nonbacteremic community-acquired pneumonia (CAP), a Luminex technology-based multiplex urinary antigen detection (UAD) diagnostic assay was developed and validated. The UAD assay can simultaneously detect 13 different serotypes of Streptococcus pneumoniae by capturing serotype-specific S. pneumoniae polysaccharides (PnPSs) secreted in human urine. Assay specificity is achieved by capturing the polysaccharides with s...

  10. Inhibition of Streptococcus pneumoniae adherence to human epithelial cells in vitro by the probiotic Lactobacillus rhamnosus GG

    Wong, SS; Quan Toh, Z; Dunne, EM; Mulholland, EK; Tang, ML; Robins-Browne, RM; Licciardi, PV; Satzke, C.

    2013-01-01

    BACKGROUND Colonization of the nasopharynx by Streptococcus pneumoniae is considered a prerequisite for pneumococcal infections such as pneumonia and otitis media. Probiotic bacteria can influence disease outcomes through various mechanisms, including inhibition of pathogen colonization. Here, we examine the effect of the probiotic Lactobacillus rhamnosus GG (LGG) on S. pneumoniae colonization of human epithelial cells using an in vitro model. We investigated the effects of LGG administered b...

  11. Morphine Disrupts Interleukin-23 (IL-23)/IL-17-Mediated Pulmonary Mucosal Host Defense against Streptococcus pneumoniae Infection▿

    Ma, Jing; Wang, Jinghua; Wan, Jing; Charboneau, Richard; Chang, Yaping; Barke, Roderick A.; Roy, Sabita

    2009-01-01

    Streptococcus pneumoniae is a pathogen that causes serious respiratory disease and meningitis in the immunocompromised drug abuse population. However, the precise mechanisms by which drug abuse compromises the host immune defense to pulmonary S. pneumoniae infection is not fully understood. Using a well-established murine model of opiate abuse and S. pneumoniae lung infection, we explored the influence of morphine treatment on the interleukin-23 (IL-23)/IL-17 axis and related innate immunity....

  12. C3b/iC3b deposition on Streptococcus pneumoniae is not affected by HIV infection

    Catherine Hyams; Jerry C H Tam; Brown, Jeremy S.; Gordon, Stephen B

    2010-01-01

    Streptococcus pneumoniae is a common cause of infection in both HIV positive patients and those with complement deficiencies. We hypothesised that HIV positive individuals might exhibit reduced opsonisation of pneumococcus with complement due to reduced levels of S. pneumoniae specific IgG. We discovered no difference in C3 deposition on S. pneumoniae between HIV positive or negative individuals, and furthermore C3 deposition remained unchanged as HIV progressed towards AIDS. We found no corr...

  13. Streptococcus pneumoniae as an agent of nosocomial infection: treatment in the era of penicillin-resistant strains

    F. Paradisi; Corti, G.; R. Cinelli

    2001-01-01

    Abstract. Streptococcus pneumoniae is a well-known agent of community-acquired infections such as sinusitis, otitis media, pneumonia, bacterial meningitis, bacteremia, and acute exacerbations of chronic bronchitis. However, the role of S.pneumoniae as a cause of nosocomial infections of respiratory tract, bloodstream, and central nervous system is more and more recognised, primarily in high-risk patients with depression of their immune function. Therapy of pneumococcal infections is made diff...

  14. Molecular Characterization of Equine Isolates of Streptococcus pneumoniae: Natural Disruption of Genes Encoding the Virulence Factors Pneumolysin and Autolysin

    Whatmore, Adrian M.; King, Samantha J.; Doherty, Neil C.; Sturgeon, Daniel; Chanter, Neil; Dowson, Christopher G.

    1999-01-01

    Although often considered a strict human pathogen, Streptococcus pneumoniae has been reported to infect and cause pneumonia in horses, although the pathology appears restricted compared to that of human infections. Here we report on the molecular characterization of a group of S. pneumoniae isolates obtained from horses in England and Ireland. Despite being obtained from geographically distinct locations, the isolates were found to represent a tight clonal group, virtually identical to each o...

  15. Macrolide resistance determinants among Streptococcus pneumoniae isolates from carriers in Central Greece

    Grivea Ioanna N

    2012-10-01

    Full Text Available Abstract Background We sought to characterize the temporal trends in nasopharyngeal carriage of macrolide-resistant pneumococci during a period with increased heptavalent pneumococcal conjugate vaccine (PCV7 coverage in Central Greece. Methods Streptococcus pneumoniae isolates were recovered from 2649 nasopharyngeal samples obtained from day-care center attendees in Central Greece during 2005–2009. A phenotypic and genotypic analysis of the isolates was performed, including the identification of macrolide resistance genes mef(A, subclasses mef(A and mef(E, as well as erm(B. Results Of the 1105 typeable S. pneumoniae isolates, 265 (24% were macrolide-resistant; 22% in 2005, 33.3% in 2006, 23.7% in 2007, and 20.5% in 2009 (P=0.398. Among these macrolide-resistant pneumococci, 28.5% possessed erm(B, 24.3% erm(B+mef(E, 41.8% mef(E, and 5.3% mef(A. A mef gene as the sole resistance determinant was carried by 31% of macrolide-resistant isolates belonging to PCV7 serotypes and 75.8% of the non-PCV7 serotypes. Across the 4 annual surveillances, pneumococci carrying mef(A gradually disappeared, whereas serotype 19F isolates carrying both erm(B and mef(E persisted without significant yearly fluctuations. Among isolates belonging to non-PCV7 serotypes, macrolide-resistance was observed in those of serotypes 6A, 19A, 10A, 15A, 15B/C, 35F, 35A, and 24F. In 2009, ie 5 years after the introduction of PCV7 in our country, 59% of macrolide-resistant pneumococci belonged to non-PCV7 serotypes. Conclusions Across the study period, the annual frequency of macrolide-resistant isolates did not change significantly, but in 2009 a marked shift to non-PCV7 serotypes occurred. Overall, more than half of the macrolide-resistant isolates possessed erm(B either alone or in combination with mef(E. erm(B dominated among isolates belonging to PCV7 serotypes, but not among those of non-PCV7 serotypes.

  16. A reação em cadeia da polimerase na detecção da resistência à penicilina em Streptococcus pneumoniae Polymerase chain reaction used to detect Streptococcus pneumoniae resistance to penicillin

    Eduardo Walker Zettler

    2004-12-01

    Full Text Available INTRODUÇÃO: O Streptococcus pneumoniae é o mais freqüente agente etiológico de infecções respiratórias adquiridas na comunidade e sua resistência aos antimicrobianos tem aumentado nos últimos anos. A determinação da resistência é feita rotineiramente por método lento que depende do crescimento em cultura e determinação da concentração inibitória mínima (CIM. A reação em cadeia da polimerase (PCR detecta os genes responsáveis pela resistência do Streptococcus pneumoniae a penicilina em cerca de 8 horas. OBJETIVO: Comparar a PCR com o método da CIM no diagnóstico da resistência da Streptococcus pneumoniae a penicilina. MÉTODO: Foram estudadas 153 amostras de Streptococcus pneumoniae, isoladas de diferentes sítios anatômicos, usando-se para detecção de mutações nos genes que codificam as proteínas ligadoras de penicilina 1a, 2b e 2x, responsáveis pela resistência à penicilina. A ocorrência das mutações foi correlacionada com a CIM de penicilina, determinada pelo teste de difusão em ágar. RESULTADOS: A resistência global à penicilina do Streptococcus pneumoniae foi de 22,8% (16,3% de resistência intermediária e 6,5% de resistência alta. Em proporções estatisticamente significativas, as amostras sensíveis à penicilina não tinham mutações, as intermediárias apenas uma, geralmente na proteína ligadora de penicilina 2x, e as altamente resistentes tinham mutações nas três proteínas investigadas. CONCLUSÃO: A PCR é um método rápido para a detecção da resistência à penicilina do Streptococcus pneumoniae, que poderá vir a ser utilizado na prática clínica.BACKGROUND: Streptococcus pneumoniae is the most common etiologic agent of community-acquired respiratory infections. In recent years, S. pneumoniae resistance to antimicrobial agents has increased. Minimum inhibitory concentration (MIC is routinely used to determine resistance. Polymerase chain reaction (PCR detects the genes

  17. PREVALENCE OF DRUG-RESISTANT STRAINS OF STREPTOCOCCUS PNEUMONIAE IN ABAKALIKI

    Iroha Ifeanyichukwu Romanus

    2012-01-01

    Full Text Available Streptococcus pneumoniae is a major cause of illness such as pneumonia, meningitis, bacteremia and otitis media in children and the elderly. The emergence of drug-resistant strains threatens to complicate the management of these diseases. We conducted a hospital-based surveillance for drug-resistant Strep. Pneumoniae in outpatients with pneumococcal infection in Abakaliki Ebonyi State Nigeria. Between 2003-2005 from January through December, a cross-sectional study was conducted in Ebonyi State University Teaching Hospital (EBSUTH Abakaliki to assess the prevalence of drug resistant strains of Streptococcus pneumoniae isolated from sputum samples of patients with pneumococcal infections attending the outpatient clinics. A total of 305 sputum samples of patients with clinically diagnosed pneumonia were collected and inoculated on 5% sheep-blood agar, incubated at 35°C for 24 h in 5-10% CO2. Colonies were Gram-stained; alpha-hemolysis colonies were tested with a 6mm optochin disk followed by bile solubility test. Susceptibility testing panels of the following antibiotics: penicillin, ciprofloxacin, clavulanic acid/amoxicillin, septrin, erythromycin, gentamycin, clarithromycin, cefotaxime and cefuroxime were tested against isolated strains of Streptococcus pneumoniae using disc diffusion method. Of the 305 sputum samples collected 30 (9.8% of S. pneumoniae was isolated from patients within the age range of 20-40 years, 140 (45.9% from patients within 41-60, 135(44.2% from patients within 60 years and above. Susceptibility studies showed that the highest resistance was 182 (59.6%, for penicillin followed by septrin 156 (51.2%, erythromycin 120(39.3%, clavulanic/amoxicillin 118(38.7% cefotaxime 114(37.4% clarithromycin 100(32.7%, ciprofloxacin 94(30.8%, gentamycin 75(24.6%, cefuroxime 70(22.9% and ceftriaxone 69(22.6%. The prevalence of S. pneumoniae resistance was relatively high and we suggest that proper antibiotics use should be adopted to avert

  18. A type IV pilus mediates DNA binding during natural transformation in Streptococcus pneumoniae.

    Raphaël Laurenceau

    Full Text Available Natural genetic transformation is widely distributed in bacteria and generally occurs during a genetically programmed differentiated state called competence. This process promotes genome plasticity and adaptability in Gram-negative and Gram-positive bacteria. Transformation requires the binding and internalization of exogenous DNA, the mechanisms of which are unclear. Here, we report the discovery of a transformation pilus at the surface of competent Streptococcus pneumoniae cells. This Type IV-like pilus, which is primarily composed of the ComGC pilin, is required for transformation. We provide evidence that it directly binds DNA and propose that the transformation pilus is the primary DNA receptor on the bacterial cell during transformation in S. pneumoniae. Being a central component of the transformation apparatus, the transformation pilus enables S. pneumoniae, a major Gram-positive human pathogen, to acquire resistance to antibiotics and to escape vaccines through the binding and incorporation of new genetic material.

  19. A Case of Necrotizing Fasciitis due to Streptococcus pneumoniae Serotype 5 in Saskatchewan

    Meenakshi Dawar

    2008-01-01

    Full Text Available Necrotizing fasciitis due to Streptococcus pneumoniae is a rare and grave condition, and only a few cases have been reported. Suggested risk factors include minor trauma, systemic lupus erythematosus, immunosuppression secondary to medication, use of intramuscular anti-inflammatories and alcoholism. A fatal case of pneumococcal necrotizing fasciitis that occurred in a 51-year-old woman with a history of alcohol abuse and oral anti-inflammatory use is presented. Her condition was caused by a multi-etiology outbreak of community-acquired pneumonia, from which S pneumoniae serotype 5 was also isolated. The case description outlines the subtle presentation and rapid clinical progression of this condition. Because serotype 5 antigen is included in the polysaccharide 23-valent pneumococcal vaccine, the present case highlights the importance of pneumococcal immunization programs in Canada.

  20. Spontaneous meningitis due to Streptococcus salivarius subsp. salivarius: cross-reaction in an assay with a rapid diagnostic kit that detected Streptococcus pneumoniae antigens.

    Shirokawa, Taijiro; Nakajima, Jun; Hirose, Kazuhito; Suzuki, Hiromichi; Nagaoka, Shoko; Suzuki, Masatsune

    2014-01-01

    Streptococcus salivarius subsp. salivarius occasionally causes meningitis associated with iatrogenic or traumatic events. We herein describe a case of meningitis caused by this organism in a patient without any apparent risk factors. In an assay of the patient's cerebrospinal fluid, cross-reaction occurred with Streptococcus pneumoniae antigen-coated latex particles in the Pastorex Meningitis Kit. In the in vitro assays, three of the five clinically isolated S. salivarius strains showed cross-reactions with the kit, indicating that these strains expressed pneumococcal antigen-like antigens. This case shows that meningitis caused by S. salivarius can occur spontaneously and it may sometimes be misdiagnosed as S. pneumoniae infection. PMID:24492701

  1. Reporting emerging resistance of Streptococcus pneumoniae from India

    Chawla Kiran

    2010-01-01

    Full Text Available Background: There are reports of emergence of resistant strains of S. pneumoniae showing resistance to penicillin from all over the world, and now, resistance to multiple drugs (multidrug-resistant strains has been added to it. However, scanty reports are available so far from India, depicting such resistance. Aims: The aim of the present study is to look for the prevalence of penicillin-resistant pneumococci and also the multidrug-resistant strains among S. pneumoniae, isolated from respiratory specimens, in the coastal part of South India. Settings and Design: A cross-sectional study was conducted from June 2008 to December 2008, in our tertiary care center. Fifty pathogenic clinical isolates were collected from patients suffering from lower respiratory tract infections. Materials and Methods: Penicillin resistance was screened by 1 µg oxacillin disk on Muller-Hinton blood agar followed by Minimum Inhibitory Concentration (MIC detection by the agar dilution method according to the Clinical Laboratory Standards Institute (CLSI guidelines. Antibiotic susceptibility for other antibiotics was carried out by the Kirby Bauer disk diffusion method followed by an E-test with HiComb test strips from Hi-media. Results: Out of 50 isolates, 4% (95% Confidence Interval - 1.4, 9.4 showed total resistance to penicillin, whereas, 10% (95% CI; 1.6, 18.3 showed intermediate resistance. These penicillin-resistant pneumococci (4% were also found to be multidrug-resistant (MDR strains. Maximum resistance was observed for cotrimoxazole and tetracycline (24% each with 95% CI; 12.2, 35.8 followed by erythromycin and ciprofloxacin (14% each with 95%CI; 4.4, 23.6. Conclusions : Increasing emergence of the resistant strains of S. pneumoniae in the community set up requires continuous monitoring and a restricted use of antibiotics to keep a check on its resistance pattern, for an effective treatment plan.

  2. Multiplex PCR to determine Streptococcus pneumoniae serotypes causing otitis media in the Republic of Ireland with further characterisation of antimicrobial susceptibilities and genotypes.

    Vickers, I

    2011-03-01

    The purpose of this study was to determine the serotypes, genotypes and antimicrobial susceptibilities of Streptococcus pneumoniae causing otitis media (OM) in children in Dublin, Ireland. S. pneumoniae isolates (n = 28) from spontaneously discharging OM were studied. Serotyping was performed using a previously undescribed multiplex polymerase chain reaction (PCR) scheme in combination with serological methods. Multilocus sequence typing (MLST) was performed using standard procedures. Antimicrobial susceptibility testing was performed using the Etest method. Fourteen different S. pneumoniae serotypes were identified. The five most common serotypes were 3, 19F, 19A, 14 and 6A, which accounted for 68% of all infections. The 7-valent pneumococcal conjugate vaccine (PCV7), 10-valent pneumococcal conjugate vaccine (PHiD-CV) and 13-valent pneumococcal conjugate vaccine (PCV13) provided potential coverages of 43%, 46% and 86%, respectively. Reduced susceptibility to penicillin was evident for 25% of isolates and was associated with serotypes 14, 19A, 19F and 9V. A total of 21 different sequence types (STs) were identified. Pneumococcal Molecular Epidemiology Network (PMEN) clones or their variants represented 54% (15\\/28) of all isolates. Continued monitoring and characterisation of S. pneumoniae causing OM in Ireland is warranted in order to guide future vaccine and treatment policies.

  3. Genome-wide identification of genes essential for the survival of Streptococcus pneumoniae in human saliva.

    Lilly M Verhagen

    Full Text Available Since Streptococcus pneumoniae transmits through droplet spread, this respiratory tract pathogen may be able to survive in saliva. Here, we show that saliva supports survival of clinically relevant S. pneumoniae strains for more than 24 h in a capsule-independent manner. Moreover, saliva induced growth of S. pneumoniae in growth-permissive conditions, suggesting that S. pneumoniae is well adapted for uptake of nutrients from this bodily fluid. By using Tn-seq, a method for genome-wide negative selection screening, we identified 147 genes potentially required for growth and survival of S. pneumoniae in saliva, among which genes predicted to be involved in cell envelope biosynthesis, cell transport, amino acid metabolism, and stress response predominated. The Tn-seq findings were validated by testing a panel of directed gene deletion mutants for their ability to survive in saliva under two testing conditions: at room temperature without CO2, representing transmission, and at 37 °C with CO2, representing in-host carriage. These validation experiments confirmed that the plsX gene and the amiACDEF and aroDEBC operons, involved in respectively fatty acid metabolism, oligopeptide transport, and biosynthesis of aromatic amino acids play an important role in the growth and survival of S. pneumoniae in saliva at 37 °C. In conclusion, this study shows that S. pneumoniae is well-adapted for growth and survival in human saliva and provides a genome-wide list of genes potentially involved in adaptation. This notion supports earlier evidence that S. pneumoniae can use human saliva as a vector for transmission.

  4. Spontaneous bacterial peritonitis due to streptococcus pneumoniae--case report.

    Litarski, Andrzej; Janczak, Dariusz; Cianciara, Jan; Merenda, Marcin

    2011-05-01

    Spontaneous bacterial peritonitis is caused by infection of ascitic fluid without any apparent intraabdominal source of infection. The disease most commonly occurs in patients with cirrhosis and 70% of cases of infections are caused by pathogenes from gastrointestinal tract. The article presents the case of 38-year-old patient with spontaneous peritonitis who was treated surgically. The primary nature of the disease was confirmed by laparotomy and bacteriological examination results (Streptoccocus pneumonia) of ascitic fluid. After 54 days of hospitalisation and undergoing re-laparotomy, he was discharged in good condition. PMID:22166482

  5. Serotype Distribution and Antimicrobial Resistance of Streptococcus pneumoniae Isolates Causing Invasive and Noninvasive Pneumococcal Diseases in Korea from 2008 to 2014

    Bae, Il Kwon; Park, Dongchul; Kim, Na Young; Song, Sae Am; Urm, Sang-Hwa; Shin, Jeong Hwan

    2016-01-01

    Introduction. Streptococcus pneumoniae is an important pathogen with high morbidity and mortality rates. The aim of this study was to evaluate the distribution of common serotypes and antimicrobial susceptibility of S. pneumoniae in Korea. Methods. A total of 378 pneumococcal isolates were collected from 2008 through 2014. We analyzed the serotype and antimicrobial susceptibility for both invasive and noninvasive isolates. Results. Over the 7 years, 3 (13.5%), 35 (10.8%), 19A (9.0%), 19F (6.6%), 6A (6.1%), and 34 (5.6%) were common serotypes/serogroups. The vaccine coverage rates of PCV7, PCV10, PCV13, and PPSV23 were 21.4%, 23.3%, 51.9%, and 62.4% in all periods. The proportions of serotypes 19A and 19F decreased and nonvaccine serotypes increased between 2008 and 2010 and 2011 and 2014. Of 378 S. pneumoniae isolates, 131 (34.7%) were multidrug resistant (MDR) and serotypes 19A and 19F were predominant. The resistance rate to levofloxacin was significantly increased (7.2%). Conclusion. We found changes of pneumococcal serotype and antimicrobial susceptibility during the 7 years after introduction of the first pneumococcal vaccine. It is important to continuously monitor pneumococcal serotypes and their susceptibilities. PMID:27314035

  6. Silica desiccant packets for storage and transport of Streptococcus pneumoniae and other clinically relevant species.

    Casey L Pell

    Full Text Available Bacterial isolates are often transported between laboratories for research and diagnostic purposes. Silica desiccant packets (SDPs, which are inexpensive and do not require freezing, were evaluated for storage and recovery of bacterial isolates. Conditions such as inoculum size, swab type and temperature of storage were investigated using ten Streptococcus pneumoniae isolates. The optimized protocol was then tested using 49 additional S. pneumoniae isolates representing 40 serogroups. Overall, S. pneumoniae growth was considered satisfactory (>100 colony forming units for 98/109 (89.9% and 20/20 (100% swabs after 14 days at room temperature or 28 days at 4° C, respectively. Storage in SDPs did not impact on the ability of S. pneumoniae isolates to be subsequently serotyped. When the survival of nine other clinically relevant bacterial species was tested, seven were viable after 28 days at room temperature, the exceptions being Neisseria gonorrhoeae and Haemophilus influenzae. SDPs are suitable for transport and short-term storage of bacterial species including S. pneumoniae.

  7. Streptococcus pneumoniae sepsis as the initial presentation of systemic lupus erythematosus

    Erdem I

    2016-09-01

    Full Text Available Ilknur Erdem,1 Senay Elbasan Omar,1 Ridvan Kara Ali,1 Hayati Gunes,2 Aynur Eren Topkaya2 1Department of Infectious Diseases, 2Department of Medical Microbiology, Faculty of Medicine, Namik Kemal University, Tekirdag, Turkey Objective: Infections are among the most important causes of morbidity and mortality in patients with systemic lupus erythematosus (SLE but are rare initial presentation of the disease. Therefore, in this study, we describe a case of Streptococcus pneumoniae sepsis in a young woman with previously undiagnosed SLE. Case report: A 23-year-old female patient was admitted to our outpatient clinic complaining of high fever (40°C, chills, fatigue, generalized myalgia, and cough with brown sputum for 5 days. Blood cultures grew gram-positive coccus defined as S. pneumoniae using standard procedures. Antinuclear antibody was positive at a titer of 1/1,000, and anti-double-stranded DNA was positive at 984 IU/mL. She was diagnosed with SLE. Her respiratory symptoms and pleural effusion were considered to be due to pulmonary manifestation of SLE. Conclusion: The underlying immunosuppression caused by SLE could have predisposed the patient to invasive pneumococcal disease. It may also occur as a primary presenting feature, although a rare condition. Keywords: Streptococcus pneumoniae, sepsis, systemic lupus erythematosus

  8. Streptococcus pneumoniae induces pyroptosis through the regulation of autophagy in murine microglia.

    Kim, Ji-Yun; Paton, James C; Briles, David E; Rhee, Dong-Kwon; Pyo, Suhkneung

    2015-12-29

    Streptococcus pneumoniae is responsible for significant mortality and morbidity worldwide and causes invasive pneumococcal diseases including pneumococcal meningitis. Pyroptosis is caspase-1-dependent inflammatory cell death and is known to be induced by various microbial infections. In the present study, we investigated the molecular mechanisms that regulate pyroptosis induced by S. pneumoniae in microglia. Our results revealed that S. pneumoniae induced pyroptosis through caspase-1 activation and IL-1β production. We also found that the activation of caspase-1 and the maturation of IL-1β and IL-18 in the S. pneumoniae-triggered pyroptotic cell death process were mediated by NLRP3 inflammasome. In addition, pneumococcal infection increased the expression of autophagy-related genes and induced autophagosome formation. We also showed that the inhibition of autophagy promoted pneumococcus-induced pyroptosis. Furthermore, ROS was generated by pneumococcal infection and inhibited caspase-1 activation within 4 h of infection. However, in the late phase of infection, IL-1β secretion and caspase-1-dependent cell death were induced by ROS. These results suggest that autophagy induction transiently delay pyroptosis induced by S. pneumoniae in microglia. Our study also revealed that the activation of caspase-1 and the production of IL-1β were induced by pneumolysin and that pneumolysin triggered pyroptosis in microglial cells. Similar to the in vitro results, S. pneumoniae induced caspase-1 activation and caspase-1-dependent cytokine maturation in the mouse meningitis model. Thus, the present data demonstrate that S. pneumoniae induces pyroptosis in murine microglia and that NLRP3 inflammasome is critical for caspase-1 activation during the process. Furthermore, the induction of autophagy could transiently protect microglia from pyroptosis. PMID:26683708

  9. Streptococcus pneumoniae capsule determines disease severity in experimental pneumococcal meningitis.

    Hathaway, Lucy J; Grandgirard, Denis; Valente, Luca G; Täuber, Martin G; Leib, Stephen L

    2016-03-01

    Streptococcus pneumoniaebacteria can be characterized into over 90 serotypes according to the composition of their polysaccharide capsules. Some serotypes are common in nasopharyngeal carriage whereas others are associated with invasive disease, but when carriage serotypes do invade disease is often particularly severe. It is unknown whether disease severity is due directly to the capsule type or to other virulence factors. Here, we used a clinical pneumococcal isolate and its capsule-switch mutants to determine the effect of capsule, in isolation from the genetic background, on severity of meningitis in an infant rat model. We found that possession of a capsule was essential for causing meningitis. Serotype 6B caused significantly more mortality than 7F and this correlated with increased capsule thickness in the cerebrospinal fluid (CSF), a stronger inflammatory cytokine response in the CSF and ultimately more cortical brain damage. We conclude that capsule type has a direct effect on meningitis severity. This is an important consideration in the current era of vaccination targeting a subset of capsule types that causes serotype replacement. PMID:27009189

  10. Novel molecular method for identification of Streptococcus pneumoniae applicable to clinical microbiology and 16S rRNA sequence-based microbiome studies

    Scholz, Christian F. P.; Poulsen, Knud; Kilian, Mogens

    2012-01-01

    The close phylogenetic relationship of the important pathogen Streptococcus pneumoniae and several species of commensal streptococci, particularly Streptococcus mitis and Streptococcus pseudopneumoniae, and the recently demonstrated sharing of genes and phenotypic traits previously considered spe...... occurred in previous reports and in reference sequence databases. Copyright © 2012 American Society for Microbiology. All Rights Reserved....

  11. Antithrombin inhibits bronchoalveolar activation of coagulation and limits lung injury during Streptococcus pneumoniae pneumonia in rats

    Choi, Goda; Hofstra, Jorrit-Jan H; Roelofs, Joris J T H; Rijneveld, Anita W; Bresser, Paul; van der Zee, Jaring S; Florquin, Sandrine; van der Poll, Tom; Levi, Marcel; Schultz, Marcus J

    2008-01-01

    OBJECTIVE: Alveolar fibrin deposition is a hallmark of pneumonia. It has been proposed that natural inhibitors of coagulation, including activated protein C, antithrombin, and tissue factor pathway inhibitor, exert lung-protective effects via anticoagulant and possibly anti-inflammatory pathways. We

  12. In Vivo Activities of Amoxicillin and Amoxicillin-Clavulanate against Streptococcus pneumoniae: Application to Breakpoint Determinations

    Andes, D.; Craig, W. A.

    1998-01-01

    The in vivo activities of amoxicillin and amoxicillin-clavulanate against 17 strains of Streptococcus pneumoniae with penicillin MICs of 0.12–8.0 mg/liter were assessed in a cyclophosphamide-induced neutropenic murine thigh infection model. Renal impairment was produced by administration of uranyl nitrate to prolong the amoxicillin half-life in the mice from 21 to 65 min, simulating human pharmacokinetics. Two hours after thigh infection with 105 to 106 CFU, groups of mice were treated with 7...

  13. Artritis séptica por streptococcus pneumoniae: reporte de un caso

    Novoa, C.; López, R.E.; Gómez Balboa, P.; Blas Dobón, J.A.; Bonet, B.; Rodrigo Pérez, José Luís

    2015-01-01

    La artritis neumocócica es infrecuente en adultos, predomina en pacientes pediátricos. La neumonía y la bacteriemia son las manifestaciones más frecuentes, la afectación articular según series recientes tiene una prevalencia menor al 1% en sujetos menores de 50 años. Se presenta el caso de una paciente de 60 años con LES (Lupus Eritematoso Sistémico) la cual presentó monoartritis séptica de rodilla derecha causada por Streptococcus pneumoniae

  14. Activities of Cethromycin and Telithromycin against Recent North American Isolates of Streptococcus pneumoniae

    Jorgensen, James H.; Crawford, Sharon A.; McElmeel, M. Leticia; Whitney, Cynthia G.

    2004-01-01

    The in vitro activities of two investigational ketolides, cethromycin (formerly ABT-773) and telithromycin, were determined for a selected group of 312 Streptococcus pneumoniae isolates from a national surveillance program. The MIC of cethromycin at which 50% of the isolates were inhibited was 0.008 μg/ml, and the MIC at which 90% of the isolates were inhibited was 0.06 μg/ml; the corresponding values for telithromycin were ≤0.015 and 0.25 μg/ml, respectively. For six quinupristin-dalfopristi...

  15. Macrolide resistance determinants in erythromycin-resistant Streptococcus pneumoniae in Turkey.

    Gulay, Zeynep; Ozbek, Ozgen Alpay; Bicmen, Meral; Gur, Deniz

    2008-11-01

    To determine the major molecular mechanisms of macrolide resistance among Streptococcus pneumoniae isolates in Turkey, we examined a total of 151 isolates collected from different regions of Turkey. Overall, 40 (26.4%) isolates were resistant to erythromycin. The most common mechanism (38/40) was target site modification due to erm (B) genes. Only two isolates harbored the mef (A)/(E) efflux gene. A clonal spread of resistant strains could not be demonstrated by BOX-polymerase chain reaction. The results from this study have shown that the erm (B) gene is predominant in Turkish S. pneumoniae isolates, as in isolates from the rest of the world, and a clonal dissemination is not responsible for this resistance profile. PMID:19050364

  16. Resistencia a penicilina y otros antimicrobianos en 103 aislamientos clínicos de Streptococcus pneumoniae (2000-2001 Resistance to penicillin and other antimicrobials in 103 clinical isolations of Streptococcus pneumoniae (2000-2001

    J.J. García-Irure

    2003-04-01

    Full Text Available Fundamentos. Conocer en nuestro hospital la sensibilidad a penicilina de aislamientos de Streptococcus pneumoniae, así como analizar la asociación de resistencia a penicilina y otros antimicrobianos y la actividad de cefotaxima y cefepima en cepas de Streptococcus pneumoniae resistentes a penicilina. Métodos. Se determinó la sensibilidad de 103 aislamientos de Streptococcus pneumoniae, procedentes de muestras clínicas durante los años 2000-2001, a penicilina, eritromicina, cloramfenicol, tetraciclina, cotrimoxazol, cefotaxima, cefepima y levofloxacino. Resultados. El 68% de los aislamientos fueron sensibles a penicilina, mientras que un 32% de las cepas de Streptococcus pneumoniae aisladas fueron resistentes a penicilina, presentando el 7,7% resistencia de alto grado a la misma. La resistencia a eritromicina, cloramfenicol, tetraciclina, cotrimoxazol y levofloxacino fue del 38,8; 9,7; 20,4; 25,2 y 2,9% respectivamente, incrementándose a valores del 66,6; 30,3; 48,5; 72,7 y 9,1% en las 33 cepas con resistencia a penicilina. La resistencia a cefotaxima y cefepima fue del 9,7 y 10,6% respectivamente. Conclusiones. Un alto porcentaje de cepas de Streptococcus pneumoniae presentaron algún grado de resistencia a penicilina, pero con cifras menores que las presentadas en otros estudios de ámbito nacional. Asimismo, se demostró que la resistencia a penicilina se asociaba significativamente (p Background. To determine in our hospital the sensitivity of isolations of Streptococcus pneumoniae to penicillin, as well as to analyse the association of resistance to penicillin and other antimicrobials and the activity of cefotaxime and cefepime in pencillin resistant strains of Streptococcus pneumoniae. Methods. The sensitivity was determined on 103 isolations of Streptococcus pneumoniae, from clinical samples from the years 2000-2001, to penicillin, eritromycine, cloramfenicol, tetracycline, cotrimoxazol, cefotaxime, cefepime and levofloxacine

  17. Nasopharyngeal carriage rate of Streptococcus pneumoniae in Ugandan children with sickle cell disease

    Kateete David P

    2012-01-01

    Full Text Available Abstract Background Nasopharyngeal carriage of Streptococcus pneumoniae is a determinant for invasive pneumococcal disease, which often complicates homozygous sickle cell disease. Here, we determined the nasopharyngeal carriage rate of S. pneumoniae in Ugandan children with homozygous sickle cell disease, who attended the outpatient Sickle Cell Clinic at Mulago National Referral hospital in Kampala, Uganda. Results S. pneumoniae occurred in 27 of the 81 children with homozygous sickle cell disease (giving a carriage rate of 33%, 27/81. Twenty three children were previously hospitalized of whom S. pneumoniae occurred in only two (9%, 2/23, while among the 58 who were not previously hospitalized it occurred in 25 (43%, 25/58, χ2 = 8.8, p = 0.003, meaning there is an association between high carriage rate and no hospitalization. Two children previously immunized with the pneumococcal conjugate vaccine did not carry the organism. Prior antimicrobial usage was reported in 53 children (65%, 53/81. There was high resistance of pneumococci to penicillin (100%, 27/27 and trimethoprime-sulfamethoxazole (97%, 26/27, but low resistance to other antimicrobials. Of the 70 children without sickle cell disease, S. pneumoniae occurred in 38 (54%, 38/70 of whom 43 were males and 27 females (53% males, 23/43, and 56% females, 15/27. Conclusion Nasopharyngeal carriage of penicillin resistant pneumococci in Ugandan children with homozygous sickle cell disease is high. While nasopharyngeal carriage of S. pneumoniae is a determinant for invasive pneumococcal disease, pneumococcal bacteremia is reportedly low in Ugandan children with sickle cell disease. Studies on the contribution of high carriage rates to invasive pneumococcal disease in these children will be helpful. This is the first report on pneumococcal carriage rate in Ugandan children with sickle cell disease.

  18. Spatial exploration of Streptococcus pneumoniae clonal clustering in São Paulo, Brazil

    Amilton Mouro

    2011-10-01

    Full Text Available OBJECTIVES: To examine the spatial distribution of Streptococcus pneumoniae and its clonal patterns collected between 2002 and 2006 in São Paulo, Brazil. METHODS: As part of an observational study in São Paulo city, Brazil, S. pneumoniae isolates routinely cultured from blood, respiratory specimens, or cerebrospinal and other profound fluids were selected. Additionally, only isolates with either penicillin (PEN intermediate (I or resistant (R status on routine antibiogram were included, in order to obtain a higher probability of clonal isolates. A single I/R S. pneumoniae isolate per patient was included and submitted to genotypic determination by pulsed field gel electrophoresis (PFGE. Minimum inhibitory concentrations (MICs were determined for the isolates by Etest® to PEN and other antimicrobials. Each isolate was geocoded in a digital map. The Kernel function and ratio methods between total isolates vs. clones were used in order to explore possible cluster formations. RESULTS: Seventy-eight (78 S. pneumoniae community isolates from two major outpatient centers in São Paulo, Brazil, were selected from the databank according to their penicillin susceptibility profile, i.e. R or I to penicillin assessed by oxacillin disc diffusion. Of these, 69 were submitted to PFGE, 65 to MIC determination, and 48 to spatial analytical procedures. Preliminary spatial analysis method showed two possible cluster formation located in southwest and southeast regions of the city. CONCLUSION: Further analyses are required for precisely determining the existence of S. pneumoniae clusters and their related risk factors. Apparently there is a specific transmission pattern of S. pneumoniae clones within certain regions and populations. GIS and spatial methods can be applied to better understand epidemiological patterns and to identify target areas for public health interventions.

  19. Effects of recombinant IL-17F intranasal inoculation against Streptococcus pneumoniae infection in a murine model.

    Chen, Ling; Guo, Sheng; Wu, Liangxia; Hao, Chunli; Xu, Wanting; Zhang, Jianhua

    2015-01-01

    Interleukin-17F (IL-17F) is an important member of IL-17 cytokine family, which plays important roles in host defense against microbial infections. Streptococcus pneumoniae is a common pathogen associated with several invasive and noninvasive pneumococcal diseases, and mucosal immune response plays crucial roles in defenses against pneumococcal infection. Thus, intranasal inoculation may be an alternative approach against pneumococci. In this study, BALB/c mice were intranasally inoculated with recombinant IL-17F (rIL-17F) prior to S. pneumoniae (American Type Culture Collection 6303, serotype 3) infection. As compared with the control group, numbers of total leukocyte, neutrophil, and macrophage in lungs were significantly increased in mice inoculated with rIL-17F. The levels of macrophage inflammatory protein 1α (MIP-1α), MIP-2β, and interferon γ were significantly increased in bronchoalveolar lavage fluid and culture supernatant of splenocytes from mice inoculated with rIL-17F. rIL-17F inoculation also significantly elevated β-defensin-2 expression in lung tissues. Furthermore, compared with S. pneumoniae infection group, rIL-17F inoculation prior to infection significantly reduced S. pneumoniae colonization in lungs. These findings demonstrated that rIL-17F intranasal inoculation strengthened host defense against pneumococci, which may be developed to prevent pneumococcal infection. PMID:25196250

  20. Gestational Hypothyroidism Improves the Ability of the Female Offspring to Clear Streptococcus pneumoniae Infection and to Recover From Pneumococcal Pneumonia.

    Nieto, Pamela A; Peñaloza, Hernán F; Salazar-Echegarai, Francisco J; Castellanos, Raquel M; Opazo, Maria Cecilia; Venegas, Luis; Padilla, Oslando; Kalergis, Alexis M; Riedel, Claudia A; Bueno, Susan M

    2016-06-01

    Maternal thyroid hormones are essential for proper fetal development. A deficit of these hormones during gestation has enduring consequences in the central nervous system of the offspring, including detrimental learning and impaired memory. Few studies have shown that thyroid hormone deficiency has a transient effect in the number of T and B cells in the offspring gestated under hypothyroidism; however, there are no studies showing whether maternal hypothyroidism during gestation impacts the response of the offspring to infections. In this study, we have evaluated whether adult mice gestated in hypothyroid mothers have an altered response to pneumococcal pneumonia. We observed that female mice gestated in hypothyroidism have increased survival rate and less bacterial dissemination to blood and brain after an intranasal challenge with Streptococcus pneumoniae. Further, these mice had higher amounts of inflammatory cells in the lungs and reduced production of cytokines characteristic of sepsis in spleen, blood, and brain at 48 hours after infection. Interestingly, mice gestated in hypothyroid mothers had basally increased vascular permeability in the lungs. These observations suggest that gestational hypothyroidism alters the immune response and the physiology of lungs in the offspring, increasing the resistance to respiratory bacterial infections. PMID:27035652

  1. Molecular Detection of Streptococcus pneumoniae on Dried Blood Spots from Febrile Nigerian Children Compared to Culture

    Iroh Tam, Pui-Ying; Hernandez-Alvarado, Nelmary; Schleiss, Mark R.; Hassan-Hanga, Fatimah; Onuchukwu, Chuma; Umoru, Dominic; Obaro, Stephen K.

    2016-01-01

    Background Nigeria has one of the highest burdens of pneumococcal disease in the world, but accurate surveillance is lacking. Molecular detection of infectious pathogens in dried blood spots (DBS) is an ideal method for surveillance of infections in resource-limited settings because of its low cost, minimal blood volumes involved, and ease of storage at ambient temperature. Our study aim was to evaluate a Streptococcus pneumoniae real-time polymerase chain reaction (rt-PCR) assay on DBS from febrile Nigerian children on Whatman 903 and FTA filter papers, compared to the gold standard of culture. Methods Between September 2011 to May 2015, blood was collected from children 5 years of age or under who presented to six hospital study sites throughout northern and central Nigeria with febrile illness, and inoculated into blood culture bottles or spotted onto Whatman 903 or FTA filter paper. Culture and rt-PCR were performed on all samples. Results A total of 537 DBS specimens from 535 children were included in the study, of which 15 were culture-positive for S. pneumoniae. The rt-PCR assay detected S. pneumoniae in 12 DBS specimens (2.2%). One positive rt-PCR result was identified in a culture-negative specimen from a high-risk subject, and two positive rt-PCR results were negative on repeat testing. Six culture-confirmed cases of S. pneumoniae bacteremia were missed. Compared to culture, the overall sensitivities of Whatman 903 and FTA DBS for detection of S. pneumoniae were 57.1% (95% CI 18.4–90.1%) and 62.5% (95% CI 24.5–91.5%), respectively. Nonspecific amplification was noted in an additional 22 DBS (4.1%). Among these, six were positive for a non-S. pneumoniae pathogen on culture. Conclusions Rt-PCR was able to detect S. pneumoniae from clinical DBS specimens, including from a culture-negative specimen. Our findings show promise of this approach as a surveillance diagnostic, but also raise important cautionary questions. Several DBS specimens were detected as

  2. Inhibition activity of wild berry juice fractions against Streptococcus pneumoniae binding to human bronchial cells

    Huttunen, Sanna; Toivanen, Marko; Arkko, Satu; Ruponen, Marika; Tikkanen-Kaukanen, Carina

    2010-01-01

    Abstract Bacterial adhesion to the cell surface is a crucial step before infection can take place. Inhibition of bacterial binding offers a novel preventive approach against infections. Cranberry (Vaccinium macrocarpon Ait.) juice has been found to have anti-adhesive activity against different bacteria. Streptococcus pneumoniae is an important pathogen and the most common cause for pneumonia, meningitis, and otitis media. In this study the inhibitory activity of cranberry (Vacciniu...

  3. Human C-Reactive Protein Protects Mice from Streptococcus pneumoniae Infection without Binding to Pneumococcal C-Polysaccharide1

    Suresh, Madathilparambil V.; Singh, Sanjay K.; Ferguson, Donald A.; Agrawal, Alok

    2007-01-01

    Human C-reactive protein (CRP) protects mice from lethality after infection with virulent Streptococcus pneumoniae type 3. For CRP-mediated protection, the complement system is required; however, the role of complement activation by CRP in the protection is not defined. Based on the in vitro properties of CRP, it has been assumed that protection of mice begins with the binding of CRP to pneumococcal C-polysaccharide on S. pneumoniae and subsequent activation of the mouse complement system. In...

  4. Mechanisms of dexamethasone-mediated inhibition of Toll-like receptor signaling induced by Neisseria meningitidis and Streptococcus pneumoniae

    Mogensen, Trine; Berg, Randi S; Paludan, Søren R;

    2008-01-01

    significantly reduces mortality and morbidity from bacterial meningitis. Here we investigate the molecular mechanisms behind the inhibitory effect of dexamethasone upon the inflammatory responses evoked by Neisseria meningitidis and Streptococcus pneumoniae, two of the major causes of bacterial meningitis. The...... and S. pneumoniae, which may contribute to our understanding of the clinical effect and the importance of timing with respect to corticosteroid treatment during bacterial meningitis. Udgivelsesdato: 2008-Jan...

  5. Surface Proteins and Pneumolysin of Encapsulated and Nonencapsulated Streptococcus pneumoniae Mediate Virulence in a Chinchilla Model of Otitis Media

    Keller, Lance E.; Bradshaw, Jessica L.; Pipkins, Haley; McDaniel, Larry S.

    2016-01-01

    Streptococcus pneumoniae infections result in a range of human diseases and are responsible for almost one million deaths annually. Pneumococcal disease is mediated in part through surface structures and an anti-phagocytic capsule. Recent studies have shown that nonencapsulated S. pneumoniae (NESp) make up a significant portion of the pneumococcal population and are able to cause disease. NESp lack some common surface proteins expressed by encapsulated pneumococci, but express surface protein...

  6. Importance of Bacterial Replication and Alveolar Macrophage-Independent Clearance Mechanisms during Early Lung Infection with Streptococcus pneumoniae

    Camberlein, Emilie; Cohen, Jonathan M.; José, Ricardo; Hyams, Catherine J.; Callard, Robin; Chimalapati, Suneeta; Yuste, Jose; Edwards, Lindsey A.; Marshall, Helina; van Rooijen, Nico; Noursadeghi, Mahdad; Brown, Jeremy S.

    2015-01-01

    Although the importance of alveolar macrophages for host immunity during early Streptococcus pneumoniae lung infection is well established, the contribution and relative importance of other innate immunity mechanisms and of bacterial factors are less clear. We have used a murine model of S. pneumoniae early lung infection with wild-type, unencapsulated, and para-amino benzoic acid auxotroph mutant TIGR4 strains to assess the effects of inoculum size, bacterial replication, capsule, and alveol...

  7. Serological and molecular capsular typing, antibiotic susceptibility and multilocus sequence typing of Streptococcus pneumoniae isolates from invasive and non-invasive infections

    ZHANG Yi-jie; CHEN Yu-shen; WANG Zhan-wei; LI Yu-qian; WANG Da-xuan; SHANG Ying; FU Rong-rong

    2013-01-01

    Background Streptococcus pneumoniae (S.pneumoniae) is a major causative agent of severe infections,including sepsis,pneumonia,meningitis,and otitis media,and has become a major public health concern.We report the pneumococcal serotype and sequence type (ST) distribution,and antimicrobial resistance of 39 S.pneumoniae strains from seven hospitals in China.Methods Blood/cerebrospinal fluid (CSF) and sputum isolates from patients were analyzed to determine S.pneumoniae serotypes by polymerase chain reaction (PCR) and the Neufeld Quellung reaction,the multilocus sequence types (MLST) by PCR and sequencing,and susceptibility to antimicrobial agents by the VITEK Gram Positive Susceptibility Card.Results A total of 39 isolates were collected including 21 blood/CSF and 18 sputum isolates.Conventional serotyping by the Quellung reaction required 749 reactions.In contrast,PCR based typing needed only 106 PCR reactions.The most frequent serotypes from the blood/CSF isolates were 14 (38.1%),19A (14.3%),23F (9.5%),and 18C (9.5%).In the sputum isolates the most frequent serotypes were 19F (33.3%),23F (16.7%),19A (11.1%),and 3 (11.1%).The incidence of penicillin resistance in the blood/CSF and sputum isolates was 66.7% and 55.6%,respectively.Statistical analysis showed that patients ≤5 years old had a higher resistance to penicillin when they compared with the patients ≥65 years old (P=0.011).Serotypes 14,19A and 19F were significantly associated with penicillin resistance (P <0.001).ST320,ST271,and ST876 isolates showed high resistant rates to several antibiotics including penicillin (P=0.006).All of the isolates of serotype 19A were resistant to both penicillin and erythromycin,and they were all multi-drug resistant (MDR) isolates.Conclusions The specificity and sensitivity of multiplexPCR are good,and this method represents a substantial savings of time and money,and can be widely used in the laboratory and clinical practice.Data from this research

  8. Feasibility and Safety of Local Treatment with Recombinant Human Tissue Factor Pathway Inhibitor in a Rat Model of Streptococcus pneumoniae Pneumonia.

    Florry E van den Boogaard

    Full Text Available Pulmonary coagulopathy is intrinsic to pulmonary injury including pneumonia. Anticoagulant strategies could benefit patients with pneumonia, but systemic administration of anticoagulant agents may lead to suboptimal local levels and may cause systemic hemorrhage. We hypothesized nebulization to provide a safer and more effective route for local administration of anticoagulants. Therefore, we aimed to examine feasibility and safety of nebulization of recombinant human tissue factor pathway inhibitor (rh-TFPI in a well-established rat model of Streptococcus (S. pneumoniae pneumonia. Thirty minutes before and every 6 hours after intratracheal instillation of S. pneumonia causing pneumonia, rats were subjected to local treatment with rh-TFPI or placebo, and sacrificed after 42 hours. Pneumonia was associated with local as well as systemic activation of coagulation. Nebulization of rh-TFPI resulted in high levels of rh-TFPI in bronchoalveolar lavage fluid, which was accompanied by an attenuation of pulmonary coagulation. Systemic rh-TFPI levels remained undetectable, and systemic TFPI activity and systemic coagulation were not affected. Histopathology revealed no bleeding in the lungs. We conclude that nebulization of rh-TFPI seems feasible and safe; local anticoagulant treatment with rh-TFPI attenuates pulmonary coagulation, while not affecting systemic coagulation in a rat model of S. pneumoniae pneumonia.

  9. Urine Streptococcus Pneumoniae Antigen in the Diagnosis of Streptococcus Pneumoniae Pneumonia Children the Significance%尿肺炎链球菌抗原在诊断儿童肺炎链球菌肺炎中的意义

    夏振雄; 杨冰; 黄靖雯

    2013-01-01

      目的:探讨尿肺炎链球菌抗原用于诊断儿童肺炎链球菌感染的临床意义。方法:收集2010年9月-2011年12月笔者所在医院住院部0~3岁呼吸道感染的患儿病例,对每位患儿同时采集尿标本和痰标本,用胶体金法测定尿肺炎链球菌抗原,对痰标本进行分离培养与鉴定,并根据结果分析。结果:尿肺炎链球菌抗原和痰培养检测差别无显著性(P>0.05)。痰培养阳性组和阴性组的尿抗原阳性率差异有显著性(P0.05).Sputum positive group and negative group of urinary antigen positive rate(P<0.01).With sputum culture for pure culture as a diagnostic streptococcus pneumoniae infection standard,colloidal gold method measuring the sensitivity of the antigen was 86.00%,specific degree was 84.96%and the accuracy was 85.25%.Conclusion:Colloidal gold method and sputum culture in the diagnosis of streptococcus pneumoniae infection may have the same effect, colloidal gold method for clinical diagnosis of streptococcus pneumoniae infection children have the certain reference significance.

  10. Moxifloxacin and Azithromycin but not Amoxicillin Protect Human Respiratory Epithelial Cells against Streptococcus pneumoniae In Vitro when Administered up to 6 Hours after Challenge

    Ulrich, Martina; Albers, Cordula; Möller, Jan-Georg; Dalhoff, Axel; Korfmann, Gisela; Künkele, Frank; Döring, Gerd

    2005-01-01

    We determined the protective effect of moxifloxacin, azithromycin, and amoxicillin against Streptococcus pneumoniae infection of respiratory cells. Moxifloxacin and azithromycin effectively killed intracellular S. pneumoniae strains and protected respiratory epithelial cells significantly even when given 6 h after S. pneumoniae challenge. Amoxicillin was less effective.

  11. Crystallization and preliminary X-ray crystallographic studies of DnaJ from Streptococcus pneumoniae

    DnaJ from Streptococcus pneumoniae (SpDnaJ) is involved in the infectious disease process and is being developed as a potential vaccine to prevent bacterial infection. Here the expression, purification, crystallization and preliminary crystallographic analysis of SpDnaJ are reported. DnaJ, cooperating with DnaK and GrpE, promotes the folding of unfolded hydrophobic polypeptides, dissociates protein complexes and translocates protein across membranes. Additionally, DnaJ from Streptococcus pneumoniae (SpDnaJ) is involved in the infectious disease process and is being developed as a potential vaccine to prevent bacterial infection. Here the expression, purification, crystallization and preliminary crystallographic analysis of SpDnaJ are reported. The crystals belong to space groups I222 or I212121 and the diffraction resolution is 3.0 Å with unit-cell parameters a = 47.68, b = 104.45, c = 234.57 Å. The crystal most likely contains one molecule in the asymmetric unit, with a VM value of 3.24 Å3 Da−1 and a solvent content of 62.1%

  12. 220D-F2 from Rubus ulmifolius kills Streptococcus pneumoniae planktonic cells and pneumococcal biofilms.

    Sharmila J Talekar

    Full Text Available Streptococcus pneumoniae (pneumococcus forms organized biofilms to persist in the human nasopharynx. This persistence allows the pneumococcus to produce severe diseases such as pneumonia, otitis media, bacteremia and meningitis that kill nearly a million children every year. While bacteremia and meningitis are mediated by planktonic pneumococci, biofilm structures are present during pneumonia and otitis media. The global emergence of S. pneumoniae strains resistant to most commonly prescribed antibiotics warrants further discovery of alternative therapeutics. The present study assessed the antimicrobial potential of a plant extract, 220D-F2, rich in ellagic acid, and ellagic acid derivatives, against S. pneumoniae planktonic cells and biofilm structures. Our studies first demonstrate that, when inoculated together with planktonic cultures, 220D-F2 inhibited the formation of pneumococcal biofilms in a dose-dependent manner. As measured by bacterial counts and a LIVE/DEAD bacterial viability assay, 100 and 200 µg/ml of 220D-F2 had significant bactericidal activity against pneumococcal planktonic cultures as early as 3 h post-inoculation. Quantitative MIC's, whether quantified by qPCR or dilution and plating, showed that 80 µg/ml of 220D-F2 completely eradicated overnight cultures of planktonic pneumococci, including antibiotic resistant strains. When preformed pneumococcal biofilms were challenged with 220D-F2, it significantly reduced the population of biofilms 3 h post-inoculation. Minimum biofilm inhibitory concentration (MBIC50 was obtained incubating biofilms with 100 µg/ml of 220D-F2 for 3 h and 6 h of incubation. 220D-F2 also significantly reduced the population of pneumococcal biofilms formed on human pharyngeal cells. Our results demonstrate potential therapeutic applications of 220D-F2 to both kill planktonic pneumococcal cells and disrupt pneumococcal biofilms.

  13. 220D-F2 from Rubus ulmifolius kills Streptococcus pneumoniae planktonic cells and pneumococcal biofilms.

    Talekar, Sharmila J; Chochua, Sopio; Nelson, Katie; Klugman, Keith P; Quave, Cassandra L; Vidal, Jorge E

    2014-01-01

    Streptococcus pneumoniae (pneumococcus) forms organized biofilms to persist in the human nasopharynx. This persistence allows the pneumococcus to produce severe diseases such as pneumonia, otitis media, bacteremia and meningitis that kill nearly a million children every year. While bacteremia and meningitis are mediated by planktonic pneumococci, biofilm structures are present during pneumonia and otitis media. The global emergence of S. pneumoniae strains resistant to most commonly prescribed antibiotics warrants further discovery of alternative therapeutics. The present study assessed the antimicrobial potential of a plant extract, 220D-F2, rich in ellagic acid, and ellagic acid derivatives, against S. pneumoniae planktonic cells and biofilm structures. Our studies first demonstrate that, when inoculated together with planktonic cultures, 220D-F2 inhibited the formation of pneumococcal biofilms in a dose-dependent manner. As measured by bacterial counts and a LIVE/DEAD bacterial viability assay, 100 and 200 µg/ml of 220D-F2 had significant bactericidal activity against pneumococcal planktonic cultures as early as 3 h post-inoculation. Quantitative MIC's, whether quantified by qPCR or dilution and plating, showed that 80 µg/ml of 220D-F2 completely eradicated overnight cultures of planktonic pneumococci, including antibiotic resistant strains. When preformed pneumococcal biofilms were challenged with 220D-F2, it significantly reduced the population of biofilms 3 h post-inoculation. Minimum biofilm inhibitory concentration (MBIC)50 was obtained incubating biofilms with 100 µg/ml of 220D-F2 for 3 h and 6 h of incubation. 220D-F2 also significantly reduced the population of pneumococcal biofilms formed on human pharyngeal cells. Our results demonstrate potential therapeutic applications of 220D-F2 to both kill planktonic pneumococcal cells and disrupt pneumococcal biofilms. PMID:24823499

  14. Emergence in France of Multiple Clones of Clinical Streptococcus pneumoniae Isolates with High-Level Resistance to Amoxicillin

    Doit, Catherine; Loukil, Chawki; Fitoussi, Frederic; Geslin, Pierre; Bingen, Edouard

    1999-01-01

    The genetic relatedness of French isolates of Streptococcus pneumoniae highly resistant to amoxicillin (MIC, ≥4 μg/ml, equal to or exceeding those of penicillin) was investigated by molecular fingerprinting. The results suggest that high-level resistance to amoxicillin has emerged within preexisting penicillin-resistant clones.

  15. Genetic Relationship between Streptococcus pneumoniae Isolates from Nasopharyngeal and Cerebrospinal Fluid of Two Infants with Pneumococcal Meningitis

    de Andrade, Ana Lucia Sampaio Sgambatti; Pimenta, Fabiana Cristina; Brandileone, Maria Cristina C.; Laval, Cristina Aparecida; Guerra, Maria Luiza; de Andrade, João Guimarães; Di Fabio, Jose Luis

    2003-01-01

    The molecular epidemiology of Streptococcus pneumoniae isolates from carriage and cerebrospinal fluid (CSF) concurrently recovered from the same individual has not yet been reported. By using pulsed-field gel electrophoresis, we demonstrated the genetic linkage among strains from CSF and nasopharynges of two children with pneumococcal meningitis. PMID:12904432

  16. SMC is recruited to oriC by ParB and promotes chromosome segregation in Streptococcus pneumoniae

    Minnen, Anita; Attaiech, Laetitia; Thon, Maria; Gruber, Stephan; Veening, Jan-Willem

    2011-01-01

    Segregation of replicated chromosomes is an essential process in all organisms. How bacteria, such as the oval-shaped human pathogen Streptococcus pneumoniae, efficiently segregate their chromosomes is poorly understood. Here we show that the pneumococcal homologue of the DNA-binding protein ParB re

  17. Structural organization of the Streptococcus pneumoniae chromosome and relatedness of penicillin-sensitive and -resistant strains in type 9V

    Gasc, AM; Giammarinaro, P; Bao, TH; Geslin, P; VanderGiezen, M; Sicard, M

    1997-01-01

    Fragmentation of Streptococcus pneumoniae genomic DNA with low-frequency-cleavage restriction endonucleases and separation of the fragments by field-inversion gel electrophoresis (FIGE) provides a DNA-fingerprint of a strain. This method enables us to construct a physical and genetic map of the R6 l

  18. Eukaryotic-type protein kinase StkP; a novel player in regulation of natural competence development in Streptococcus pneumoniae

    Přenosilová, Lenka; Nováková, Linda; Branny, Pavel

    Washington : American Society for Microbiology, 2006, s. 73-73. ISBN 1-55581-400-X. [ASM Conferences Streptococcal Genetics. Saint Malo (FR), 18.06.2006-21.06.2006] Institutional research plan: CEZ:AV0Z50200510 Keywords : stkp * Streptococcus pneumoniae Subject RIV: EE - Microbiology, Virology

  19. LocZ is a new cell division protein involved in proper septum placement in Streptococcus pneumoniae

    Holečková, Nela; Doubravová, Linda; Massidda, Orietta; Molle, Virginie; Buriánková, Karolína; Benada, Oldřich; Kofroňová, Olga; Ulrych, Aleš; Branny, Pavel

    2015-01-01

    Roč. 6, č. 1 (2015), s. 1-13. ISSN 2150-7511 R&D Projects: GA ČR GAP207/12/1568; GA ČR GAP302/12/0256 Institutional support: RVO:61388971 Keywords : cell division * septum placement * Streptococcus pneumoniae Subject RIV: EE - Microbiology, Virology Impact factor: 6.786, year: 2014

  20. Penicillin resistance and serotype distribution of Streptococcus pneumoniae in Ghanaian children less than six years of age

    Dayie, Nicholas T. K. D.; Arhin, Reuben E.; Newman, Mercy J.;

    2013-01-01

    Background: The objective of this study was to determine the prevalence of nasopharyngeal carriage, serotype distribution, and penicillin resistance of Streptococcus pneumoniae in children 2 mu g/ml and were classified as fully penicillin resistant with 45% of the isolates having intermediate res...

  1. Serotypes and Antibiotic Susceptibility of Streptococcus pneumoniae Isolates from Invasive Pneumococcal Disease and Asymptomatic Carriage in a Pre-vaccination Period, in Algeria

    Ziane, Hanifa; Manageiro, Vera; Ferreira, Eugénia; Moura, Inês B.; Bektache, Soumia; Tazir, Mohamed; Caniça, Manuela

    2016-01-01

    In Algeria, few data is available concerning the distribution of pneumococcal serotypes and respective antibiotic resistance for the current pre-vaccination period, which is a public health concern. We identified the most frequent Streptococcus pneumoniae serogroup/types implicated in invasive pneumococcal disease (IPD; n = 80) and carriage (n = 138) in Algerian children younger than 5 years old. Serogroup/types of 78 IPD isolates were identified by capsular typing using a sequential multiplex PCR. Overall, serotypes 14, 19F, 6B, 23F, 18C, 1, 5, 7F, 19A, and 3 (55% of PCV7 serotypes, 71.3% of PCV10, and 90% of PCV13) were identified. Additionally, 7.5% of the non-vaccine serotypes 6C, 9N/L, 20, 24F, 35B, and 35F, were observed. In the case of S. pneumoniae asymptomatic children carriers, the most common serogroup/types were 6B, 14, 19F, 23F, 4, 9V/A, 1, 19A, 6A, and 3 (42.7% of PCV7 serotypes, 44.2% of PCV10, and 58% of PCV13). For 6.1% of the cases co-colonization was detected. Serotypes 14, 1, 5, and 19A were more implicated in IPD (p vaccine serotypes, the rates of penicillin non-susceptible isolates were higher in no meningitis cases (80%) than in meningitis (66.7%), with serotypes 14, 19A, 19F, and 23F presenting the highest MIC levels (>2μg/ml). Resistance to cefotaxime was higher in isolates from meningitis (40.5%); however, resistance to erythromycin and co-trimoxazole (>40%) was more pronounced in no-meningeal forms. Overall, our results showed that PCV13 conjugate vaccine would cover up to 90% of the circulating isolates associated with IPD in Algeria, highlighting the importance of monitoring the frequency of S. pneumoniae serogroups/types during pre- and post-vaccination periods. PMID:27379023

  2. Crystallization of dihydrodipicolinate synthase from a clinical isolate of Streptococcus pneumoniae

    The emergence of drug-resistant bacteria highlights the importance of identifying potential drug targets. Dihydrodipicolinate synthase (DHDPS) is a valid but as yet unexploited antimicrobial target that functions in the biosynthesis of (S)-lysine. In this study, the cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DHDPS from S. pneumoniae are described. Dihydrodipicolinate synthase (DHDPS; EC 4.2.1.52) catalyzes the rate-limiting step in the (S)-lysine biosynthesis pathway of bacteria and plants. Here, the cloning of the DHDPS gene from a clinical isolate of Streptococcus pneumoniae (OXC141 strain) and the strategy used to express, purify and crystallize the recombinant enzyme are described. Diffracting crystals were grown in high-molecular-weight PEG precipitants using the hanging-drop vapour-diffusion method. The best crystal, from which data were collected, diffracted to beyond 2.0 Å resolution. Initially, the crystals were thought to belong to space group P42212, with unit-cell parameters a = 105.5, b = 105.5, c = 62.4 Å. However, the R factors remained high following initial processing of the data. It was subsequently shown that the data set was twinned and it was thus reprocessed in space group P2, resulting in a significant reduction in the R factors. Determination of the structure will provide insight into the design of novel antimicrobial agents targeting this important enzyme from S. pneumoniae

  3. Dominance of multidrug resistant CC271 clones in macrolide-resistant streptococcus pneumoniae in Arizona

    Bowers Jolene R

    2012-01-01

    Full Text Available Abstract Background Rates of resistance to macrolide antibiotics in Streptococcus pneumoniae are rising around the world due to the spread of mobile genetic elements harboring mef(E and erm(B genes and post-vaccine clonal expansion of strains that carry them. Results Characterization of 592 clinical isolates collected in Arizona over a 10 year period shows 23.6% are macrolide resistant. The largest portion of the macrolide-resistant population, 52%, is dual mef(E/erm(B-positive. All dual-positive isolates are multidrug-resistant clonal lineages of Taiwan19F-14, mostly multilocus sequence type 320, carrying the recently described transposon Tn2010. The remainder of the macrolide resistant S. pneumoniae collection includes 31% mef(E-positive, and 9% erm(B-positive strains. Conclusions The dual-positive, multidrug-resistant S. pneumoniae clones have likely expanded by switching to non-vaccine serotypes after the heptavalent pneumococcal conjugate vaccine release, and their success limits therapy options. This upsurge could have a considerable clinical impact in Arizona.

  4. Characterization of Spbhp-37, a Hemoglobin-Binding Protein of Streptococcus pneumoniae

    Romero-Espejel, María E.; Rodríguez, Mario A.; Chávez-Munguía, Bibiana; Ríos-Castro, Emmanuel; Olivares-Trejo, José de Jesús

    2016-01-01

    Streptococcus pneumoniae is a Gram-positive microorganism that is the cause of bacterial pneumonia, sinusitis and otitis media. This human pathogen also can cause invasive diseases such as meningitis, bacteremia and septicemia. Hemoglobin (Hb) and haem can support the growth and viability of S. pneumoniae as sole iron sources. Unfortunately, the acquisition mechanism of Hb and haem in this bacterium has been poorly studied. Previously we identified two proteins of 37 and 22 kDa as putative Hb- and haem-binding proteins (Spbhp-37 and Spbhp-22, respectively). The sequence of Spbhp-37 protein was database annotated as lipoprotein without any function or localization. Here it was immunolocalized in the surface cell by transmission electron microscopy using specific antibodies produced against the recombinant protein. The expression of Spbhp-37 was increased when bacteria were grown in media culture supplied with Hb. In addition, the affinity of Sphbp-37 for Hb was determined. Thus, in this work we are presenting new findings that attempt to explain the mechanism involved in iron acquisition of this pathogen. In the future these results could help to develop new therapy targets in order to avoid the secondary effects caused by the traditional therapies. PMID:27200302

  5. Pyruvate oxidase influences the sugar utilization pattern and capsule production in Streptococcus pneumoniae.

    Sandra M Carvalho

    Full Text Available Pyruvate oxidase is a key function in the metabolism and lifestyle of many lactic acid bacteria and its activity depends on the presence of environmental oxygen. In Streptococcus pneumoniae the protein has been suggested to play a major role in metabolism and has been implicated in virulence, oxidative stress survival and death in stationary phase. Under semi-aerobic conditions, transcriptomic and metabolite profiling analysis of a spxB mutant grown on glucose showed minor changes compared to the wild type, apart from the significant induction of two operons involved in carbohydrate uptake and processing. This induction leads to a change in the sugar utilization capabilities of the bacterium, as indicated by the analysis of the growth profiles of the D39 parent and spxB mutant on alternative carbohydrates. Metabolic analysis and growth experiments showed that inactivation of SpxB has no effect on the glucose fermentation pattern, except under aerobic conditions. More importantly, we show that mutation of spxB results in the production of increased amounts of capsule, the major virulence factor of S. pneumoniae. Part of this increase can be attributed to induction of capsule operon (cps transcription. Therefore, we propose that S. pneumoniae utilizes pyruvate oxidase as an indirect sensor of the oxygenation of the environment, resulting in the adaption of its nutritional capability and the amount of capsule to survive in the host.

  6. Dried Saliva Spots: A Robust Method for Detecting Streptococcus pneumoniae Carriage by PCR

    Cassandra L. Krone

    2016-03-01

    Full Text Available The earliest studies in the late 19th century on Streptococcus pneumoniae (S. pneumoniae carriage used saliva as the primary specimen. However, interest in saliva declined after the sensitive mouse inoculation method was replaced by conventional culture, which made isolation of pneumococci from the highly polymicrobial oral cavity virtually impossible. Here, we tested the feasibility of using dried saliva spots (DSS for studies on pneumococcal carriage. Saliva samples from children and pneumococcus-spiked saliva samples from healthy adults were applied to paper, dried, and stored, with and without desiccant, at temperatures ranging from −20 to 37 °C for up to 35 days. DNA extracted from DSS was tested with quantitative-PCR (qPCR specifically for S. pneumoniae. When processed immediately after drying, the quantity of pneumococcal DNA detected in spiked DSS from adults matched the levels in freshly spiked raw saliva. Furthermore, pneumococcal DNA was stable in DSS stored with desiccant for up to one month over a broad range of temperatures. There were no differences in the results when spiking saliva with varied pneumococcal strains. The collection of saliva can be a particularly useful in surveillance studies conducted in remote settings, as it does not require trained personnel, and DSS are resilient to various transportation conditions.

  7. MicroRNAs Constitute a Negative Feedback Loop in Streptococcus pneumoniae-Induced Macrophage Activation.

    Griss, Kathrin; Bertrams, Wilhelm; Sittka-Stark, Alexandra; Seidel, Kerstin; Stielow, Christina; Hippenstiel, Stefan; Suttorp, Norbert; Eberhardt, Martin; Wilhelm, Jochen; Vera, Julio; Schmeck, Bernd

    2016-07-15

    Streptococcus pneumoniae causes high mortality as a major pneumonia-inducing pathogen. In pneumonia, control of innate immunity is necessary to prevent organ damage. We assessed the role of microRNAs (miRNAs) as regulators in pneumococcal infection of human macrophages. Exposure of primary blood-derived human macrophages with pneumococci resulted in transcriptional changes in several gene clusters and a significant deregulation of 10 microRNAs. Computational network analysis retrieved miRNA-146a as one putatively important regulator of pneumococci-induced host cell activation. Its induction depended on bacterial structural integrity and was completely inhibited by blocking Toll-like receptor 2 (TLR-2) or depleting its mediator MyD88. Furthermore, induction of miRNA-146a release did not require the autocrine feedback of interleukin 1β and tumor necrosis factor α released from infected macrophages, and it repressed the TLR-2 downstream mediators IRAK-1 and TRAF-6, as well as the inflammatory factors cyclooxygenase 2 and interleukin 1β. In summary, pneumococci recognition induces a negative feedback loop, preventing excessive inflammation via miR-146a and potentially other miRNAs. PMID:26984146

  8. Differential Recognition and Hydrolysis of Host Carbohydrate Antigens by Streptococcus pneumoniae Family 98 Glycoside Hydrolases

    Higgins, M.; Whitworth, G; El Warry, N; Randriantsoa, M; Samain, E; Burke, R; Vocadlo, D; Boraston, A

    2009-01-01

    The presence of a fucose utilization operon in the Streptococcus pneumoniae genome and its established importance in virulence indicates a reliance of this bacterium on the harvesting of host fucose-containing glycans. The identities of these glycans, however, and how they are harvested is presently unknown. The biochemical and high resolution x-ray crystallographic analysis of two family 98 glycoside hydrolases (GH98s) from distinctive forms of the fucose utilization operon that originate from different S. pneumoniae strains reveal that one enzyme, the predominant type among pneumococcal isolates, has a unique endo-{beta}-galactosidase activity on the LewisY antigen. Altered active site topography in the other species of GH98 enzyme tune its endo-{beta}-galactosidase activity to the blood group A and B antigens. Despite their different specificities, these enzymes, and by extension all family 98 glycoside hydrolases, use an inverting catalytic mechanism. Many bacterial and viral pathogens exploit host carbohydrate antigens for adherence as a precursor to colonization or infection. However, this is the first evidence of bacterial endoglycosidase enzymes that are known to play a role in virulence and are specific for distinct host carbohydrate antigens. The strain-specific distribution of two distinct types of GH98 enzymes further suggests that S. pneumoniae strains may specialize to exploit host-specific antigens that vary from host to host, a factor that may feature in whether a strain is capable of colonizing a host or establishing an invasive infection.

  9. Disease isolates of Streptococcus pseudopneumoniae and non-typeable S. pneumoniae presumptively identified as atypical S. pneumoniae in Spain.

    Dora Rolo

    Full Text Available We aimed to obtain insights on the nature of a collection of isolates presumptively identified as atypical Streptococcus pneumoniae recovered from invasive and non-invasive infections in Spain. One-hundred and thirty-two isolates were characterized by: optochin susceptibility in ambient and CO(2-enriched atmosphere; bile solubility; PCR-based assays targeting pneumococcal genes lytA, ply, pspA, cpsA, Spn9802, aliB-like ORF2, and a specific 16S rRNA region; multilocus sequence analysis; and antimicrobial susceptibility. By multilocus sequence analysis, 61 isolates were S. pseudopneumoniae, 34 were pneumococci, 13 were S. mitis, and 24 remained unclassified as non-pneumococci. Among S. pseudopneumoniae isolates, 51 (83.6% were collected from respiratory tract samples; eight isolates were obtained from sterile sources. High frequency of non-susceptibility to penicillin (60.7% and erythromycin (42.6% was found. Only 50.8% of the S. pseudopneumoniae isolates displayed the typical optochin phenotype originally described for this species. None harbored the cpsA gene or the pneumococcal typical lytA restriction fragment length polymorphism. The Spn9802 and the specific 16S rRNA regions were detected among the majority of the S. pseudopneumoniae isolates (n = 59 and n = 49, respectively. The ply and pspA genes were rarely found. A high genetic diversity was found and 59 profiles were identified. Among the S. pneumoniae, 23 were capsulated and 11 were non-typeable. Three non-typeable isolates, associated to international non-capsulated lineages, were recovered from invasive disease sources. In conclusion, half of the atypical pneumococcal clinical isolates were, in fact, S. pseudopneumoniae and one-fourth were other streptococci. We identified S. pseudopneumoniae and non-typeable pneumococci as cause of disease in Spain including invasive disease.

  10. [Detection and Serotyping of Streptococcus pneumoniae Carried in Healthy Adults with a Modified PCR Method].

    Ishihara, Yuka; Okamoto, Akira; Ohta, Michio

    2015-05-01

    Detection of Streptococcus pneumoniae colonized in the pharynx of healthy carriers currently relies on conventional culture methods of direct plating with pharyngeal swab specimens. The accurate measurement of the carriage of pneumococci, however, has not been necessarily achieved with these methods due to low density colonization and contamination of numerous oral streptococci that express α-hemolysis. A PCR-based detection method of pneumococci-specific for lytA as well as PCR serotyping of S. pneumoniae was recently developed and their effectiveness was confirmed. We modified the reaction conditions of these methods to improve the detection rate and applied them to the measurement of S. pneumoniae carried in healthy adults. Pharyngeal swab specimens obtained from 110 healthy volunteers over 40 and living in Nagoya were enriched for 5 hours with broth medium supplemented with rabbit serum and the template DNA for PCR was extracted from the mixed enriched culture. Of 110 specimens 36 (32.7%) were lytA-positive, the rate of which was much higher than the results of previous culture-based studies. The DNA template preparations were then used for PCR-based serotyping with primers specific for each of the types included in pneumococcal 23 valent vaccine (PPV23). We found that 28 out of 36 lytA-positive carriers were identified as being positive for the serotypes belonging to PPV23, although serotypes 6A and 6B were indistinguishable with the PCR method. The most frequent serotype was serotype 14, and serotypes 4, 18C, and 6A/B were also frequently identified. Five lytA-positive carriers were previously vaccinated with PPV23, and among them, 4 were positive for serotypes contained in PPV23. We recommend PCR-based identification and serotyping of S. pneumoniae in broth enrichment culture of pharyngeal swab specimens as a reliable method for the surveillance of healthy carriers with low density colonization. PMID:26552129

  11. The Oral Commensal Streptococcus mitis Shows a Mixed Memory Th Cell Signature That Is Similar to and Cross-Reactive with Streptococcus pneumoniae

    2014-01-01

    Background Carriage of and infection with Streptococcus pneumoniae is known to predominantly induce T helper 17 (Th17) responses in humans, but the types of Th cells showing reactivity towards commensal streptococci with low pathogenic potential, such as the oral commensals S. mitis and S. salivarius, remain uncharacterized. Methods Memory CD4+ T helper (Th) cell subsets were isolated from healthy human blood donors according to differential expression of chemokine receptors, expanded in v...

  12. Complete genome sequence of a serotype 11A, ST62 Streptococcus pneumoniae invasive isolate

    Superti Fabiana

    2011-02-01

    Full Text Available Abstract Background Streptococcus pneumoniae is an important human pathogen representing a major cause of morbidity and mortality worldwide. We sequenced the genome of a serotype 11A, ST62 S. pneumoniae invasive isolate (AP200, that was erythromycin-resistant due to the presence of the erm(TR determinant, and carried out analysis of the genome organization and comparison with other pneumococcal genomes. Results The genome sequence of S. pneumoniae AP200 is 2,130,580 base pair in length. The genome carries 2216 coding sequences (CDS, 56 tRNA, and 12 rRNA genes. Of the CDSs, 72.9% have a predicted biological known function. AP200 contains the pilus islet 2 and, although its phenotype corresponds to serotype 11A, it contains an 11D capsular locus. Chromosomal rearrangements resulting from a large inversion across the replication axis, and horizontal gene transfer events were observed. The chromosomal inversion is likely implicated in the rebalance of the chromosomal architecture affected by the insertions of two large exogenous elements, the erm(TR-carrying Tn1806 and a functional prophage designated ϕSpn_200. Tn1806 is 52,457 bp in size and comprises 49 ORFs. Comparative analysis of Tn1806 revealed the presence of a similar genetic element or part of it in related species such as Streptococcus pyogenes and also in the anaerobic species Finegoldia magna, Anaerococcus prevotii and Clostridium difficile. The genome of ϕSpn_200 is 35,989 bp in size and is organized in 47 ORFs grouped into five functional modules. Prophages similar to ϕSpn_200 were found in pneumococci and in other streptococcal species, showing a high degree of exchange of functional modules. ϕSpn_200 viral particles have morphologic characteristics typical of the Siphoviridae family and are capable of infecting a pneumococcal recipient strain. Conclusions The sequence of S. pneumoniae AP200 chromosome revealed a dynamic genome, characterized by chromosomal rearrangements and

  13. Differential activation of inflammatory pathways in A549 type II pneumocytes by Streptococcus pneumoniae strains with different adherence properties

    Horvat Rebecca T

    2006-04-01

    Full Text Available Abstract Background Adherence of Streptococcus pneumoniae bacteria to lung cells is a first step in the progression from asymptomatic carriage to pneumonia. Adherence abilities vary widely among S. pneumoniae patient isolates. In this study, the binding properties of S. pneumoniae isolates and the effects of binding on activation of the Nuclear Factor-Kappa-B (NFκB pathway and cytokine secretion by type II pneumocytes were measured. Methods Mechanisms of high- and low-binding S. pneumoniae adherence to A549 cells were investigated by blocking putative receptors on bacteria and host cells with antibody and by eluting choline-binding proteins off of bacterial surfaces. NFκB activation was measured by western blot and immunocytochemistry and cytokine secretion was detected by a protein array. Results This study shows that S. pneumoniae isolates from pneumonia patients (n = 298 can vary by as much as 1000-fold in their ability to bind to human lung epithelial cells. This difference resulted in differential activation of the NFκB pathway. High-, but not low-binding S. pneumoniae used Choline-binding protein A (CbpA to bind to complement component C3 on epithelial cell surfaces. Interleukin-8 (IL-8 was the only cytokine secreted by cells treated with either low- or high-binding S. pneumoniae. Conclusion These results indicate that S. pneumoniae clinical isolates are not homogeneous in their interaction with host epithelial cells. The differential activation of host cells by high- and low-binding S. pneumoniae strains could have implications for the treatment of pneumococcal pneumonia and for vaccine development.

  14. Time and dose-dependent risk of pneumococcal pneumonia following influenza: a model for within-host interaction between influenza and Streptococcus pneumoniae

    Shrestha, Sourya; Foxman, Betsy; Dawid, Suzanne; Aiello, Allison E.; Davis, Brian M.; Berus, Joshua; Rohani, Pejman

    2013-01-01

    A significant fraction of seasonal and in particular pandemic influenza deaths are attributed to secondary bacterial infections. In animal models, influenza virus predisposes hosts to severe infection with both Streptococcus pneumoniae and Staphylococcus aureus. Despite its importance, the mechanistic nature of the interaction between influenza and pneumococci, its dependence on the timing and sequence of infections as well as the clinical and epidemiological consequences remain unclear. We e...

  15. Antagonism between penicillin and erythromycin against Streptococcus pneumoniae in vitro and in vivo

    Johansen, H K; Jensen, T G; Dessau, R B;

    2000-01-01

    effect of the bactericidal agent. In this study, the possible interaction between penicillin and erythromycin was investigated in vitro and in vivo against four clinical isolates of Streptococcus pneumoniae with MICs of penicillin ranging from 0.016 to 0.5 mg/L and of erythromycin from 0. 25 to >128 mg....../L. In vitro time-kill curves were generated with clinically relevant concentrations of penicillin (10 mg/L) and erythromycin (1 mg/L), either individually or in combination. Antagonism between penicillin and erythromycin was observed for the four isolates. In vivo interaction was investigated in the...... mouse peritonitis model. After intraperitoneal inoculation, penicillin and erythromycin were given either individually or in combination. For two of the four isolates, mortality was significantly higher in the groups treated with the combination of penicillin and erythromycin than in the groups treated...

  16. Regulation of naturally acquired mucosal immunity to Streptococcus pneumoniae in healthy Malawian adults and children.

    Sarah J Glennie

    Full Text Available Worldwide, invasive pneumococcal disease caused by Streptococcus pneumoniae is most common in young children. In adults, disease rates decline following intermittent colonization and the acquisition of naturally acquired immunity. We characterized mucosal and systemic pneumococcal-specific T-cell responses in African children and adults who contend with intense rates of colonization, up to 100% and 60% respectively. We find most Malawian children have high pneumococcal-specific T-cell responses in tonsil tissue and peripheral blood. In addition, frequent commensalism generates CD25(hi (Tregs which modulate mucosal pneumococcal-specific T-cell responses in some children and ≥50% of adults. We propose that immune regulation may prolong pneumococcal colonization and predispose vulnerable individuals to disease.

  17. Antimicrobial susceptibility patterns of Streptococcus pneumoniae over 6 years at Gondar University Hospital, Northwest Ethiopia

    Belay Anagaw; Chandrashekhar Unakal; Mucheye Gezachew; Fantahun Biadgelgene; Berhanu Anagaw; Tariku Geleshe; Birke Taddese; Birhanu Getie; Mengistu Endris; Andargachew Mulu

    2013-01-01

    Objective:To assess the magnitude and antimicrobial susceptibility patterns of Streptococcus pneumoniae isolates from various clinical specimens. Methods:A record based on retrospective study was conducted at Gondar University Teaching Hospital from September 2007 to January 2012. All patients who visited Gondar University Hospital and provided clinical specimens (body fluids, discharge, swab and blood) for routine bacteriological culturing and antimicrobial susceptibility testing were taken for analysis. Clinical specimens were processed for bacterial culture according to the standard procedures. Antimicrobial susceptibility test for isolated organisms was done using agar disk diffusion method. The data were entered and analyzed using SPSS software version 16 package. Results: One hundred and fifty three Streptococcus pneumoniae were isolated from patients who visited Gondar University Teaching Hospital bacteriology laboratory for culture. Majority of the pneumococcal isolates were from inpatients [111(72.5%)], and 74(48.4%) were from body fluids. Out of the total isolates, 93(61%) were found to be resistant to at least one antibiotic used for susceptibility testing. Forty eight (43.2%) of the isolates were multi-drug resistant (resistant to two or more drugs). The resistance rate noted for both ciprofloxacin 17(11.1%) and ceftriaxone 15(9.8%) were alarming. Conclusions: High proportions of the isolates tend to be increasingly resistant to the commonly prescribed drugs. The recommended drug of choice like ciprofloxacin and ceftriaxone were found to be less susceptible in the study area. Based on the findings, we therefore recommend that antimicrobial agents should be inspected for acceptable activity before they are prescribed and administered empirically. Further study with a better design and survey of antimicrobial susceptibility at large scale shoule be performed to draw advanced information.

  18. Children from 0 to 4 Years Old Carrying Streptococcus Pneumoniae. A Community Study

    Olga Olivia Tejeda Hernández

    2011-08-01

    Full Text Available Fundamento: el Streptococcus pneumoniae es la primera causa de neumonías comunitarias; a la infección la precede la colonización de la nasofaringe, de ahí la importancia de determinar el estado de portador y los patrones de susceptibilidad y resistencia. Objetivo: determinar la prevalencia del estado de portador de Streptococcus pneumoniae entre una población infantil menor de cuatro años. Métodos: estudio de corte transversal que incluyó 179 niños, de áreas urbanas y rurales del municipio San Antonio de los Baños, provincia Artemisa. Se estratificó el municipio por el número de nacidos en cada año, se determinó el por ciento que representaban en la muestra, asignando un número de niños a cada grupo, proporcional al tamaño del grupo poblacional estratificado. Los niños se seleccionaron de forma aleatoria en los consultorios. Se tomaron muestras de exudados nasales y faríngeos. La identificación de las cepas se realizó por las características morfológicas y culturales, prueba de la opto quina y solubilidad en bilis; la confirmación del cultivo por aglutinación al látex. Se determinó la susceptibilidad antimicrobiana por el método de Kirby-Bauer. Resultados: el 20, 6 % era portador nasal y el 33, 6 % en la faringe, estos últimos mayormente en el área urbana. No hubo diferencias por sexo; el 60 % de las cepas mostraron susceptibilidad disminuida a la penicilina, 52 % de resistencia al trimetropin- sulfametoxasol y un 25 % a la eritromicina. El total de los aislamientos fue susceptible a ofloxacina, cefotaxima, azitromicina y vancomicina. Conclusiones: el estado de portador fue alto.

  19. The Small Molecule DAM Inhibitor, Pyrimidinedione, Disrupts Streptococcus pneumoniae Biofilm Growth In Vitro.

    Mukesh Kumar Yadav

    Full Text Available Streptococcus pneumoniae persist in the human nasopharynx within organized biofilms. However, expansion to other tissues may cause severe infections such as pneumonia, otitis media, bacteremia, and meningitis, especially in children and the elderly. Bacteria within biofilms possess increased tolerance to antibiotics and are able to resist host defense systems. Bacteria within biofilms exhibit different physiology, metabolism, and gene expression profiles than planktonic cells. These differences underscore the need to identify alternative therapeutic targets and novel antimicrobial compounds that are effective against pneumococcal biofilms. In bacteria, DNA adenine methyltransferase (Dam alters pathogenic gene expression and catalyzes the methylation of adenine in the DNA duplex and of macromolecules during the activated methyl cycle (AMC. In pneumococci, AMC is involved in the biosynthesis of quorum sensing molecules that regulate competence and biofilm formation. In this study, we examine the effect of a small molecule Dam inhibitor, pyrimidinedione, on Streptococcus pneumoniae biofilm formation and evaluate the changes in global gene expression within biofilms via microarray analysis. The effects of pyrimidinedione on in vitro biofilms were studied using a static microtiter plate assay, and the architecture of the biofilms was viewed using confocal and scanning electron microscopy. The cytotoxicity of pyrimidinedione was tested on a human middle ear epithelium cell line by CCK-8. In situ oligonucleotide microarray was used to compare the global gene expression of Streptococcus pneumoniae D39 within biofilms grown in the presence and absence of pyrimidinedione. Real-time RT-PCR was used to study gene expression. Pyrimidinedione inhibits pneumococcal biofilm growth in vitro in a concentration-dependent manner, but it does not inhibit planktonic cell growth. Confocal microscopy analysis revealed the absence of organized biofilms, where cell

  20. A single amino acid substitution in the MurF UDP-MurNAc-pentapeptide synthetase renders Streptococcus pneumoniae dependent on CO2 and temperature

    Burghout, Peter; Quintero, Beatriz; Bos, Linda; Beilharz, Katrin; Veening, Jan-Willem; de Jonge, Marien I.; van der Linden, Mark; van der Ende, Arie; Hermans, Peter W. M.

    2013-01-01

    The respiratory tract pathogen Streptococcus pneumoniae encounters different levels of environmental CO2 during transmission, host colonization and disease. About 8% of all pneumococcal isolates are capnophiles that require CO2-enriched growth conditions. The underlying molecular mechanism for caphn

  1. Interleukin-10 plays a key role in the modulation of neutrophils recruitment and lung inflammation during infection by Streptococcus pneumoniae.

    Peñaloza, Hernán F; Nieto, Pamela A; Muñoz-Durango, Natalia; Salazar-Echegarai, Francisco J; Torres, Javiera; Parga, María J; Alvarez-Lobos, Manuel; Riedel, Claudia A; Kalergis, Alexis M; Bueno, Susan M

    2015-09-01

    Streptococcus pneumoniae is a major aetiological agent of pneumonia worldwide, as well as otitis media, sinusitis, meningitis and sepsis. Recent reports have suggested that inflammation of lungs due to S. pneumoniae infection promotes bacterial dissemination and severe disease. However, the contribution of anti-inflammatory molecules to the pathogenesis of S. pneumoniae remains unknown. To elucidate whether the production of the anti-inflammatory cytokine interleukin-10 (IL-10) is beneficial or detrimental for the host during pneumococcal pneumonia, we performed S. pneumoniae infections in mice lacking IL-10 (IL-10(-/-) mice). The IL-10(-/-) mice showed increased mortality, higher expression of pro-inflammatory cytokines, and an exacerbated recruitment of neutrophils into the lungs after S. pneumoniae infection. However, IL-10(-/-) mice showed significantly lower bacterial loads in lungs, spleen, brain and blood, when compared with mice that produced this cytokine. Our results support the notion that production of IL-10 during S. pneumoniae infection modulates the expression of pro-inflammatory cytokines and the infiltration of neutrophils into the lungs. This feature of IL-10 is important to avoid excessive inflammation of tissues and to improve host survival, even though bacterial dissemination is less efficient in the absence of this cytokine. PMID:26032199

  2. Streptococcus pneumoniae infection in children: vaccine implications%儿童肺炎链球菌感染与疫苗

    陆敏

    2010-01-01

    肺炎链球菌是儿童呼吸道感染中最常见的病原之一,也是导致儿童重症肺炎、肺炎并发症和死亡的主要致病菌.近年来,由于世界各地肺炎链球菌对抗生素的耐药不断增加和广泛传播,造成疾病的负担日益增加,也使临床诊治面临严峻挑战.疫苗的出现和推广在肺炎链球菌病的防治方面有着光明的前景.%Streptococcus pneumoniae is one of the most common causes of respiratory tract infections in children, and also the major pathogen leading to severe pneumonia, complications and even death. In recent years, antimicrobial resistance of streptococcus pneumoniae is growing and widespread, resulting in the increasing burden of disease,and also bring serious challenges to clinical diagnosis and treatment. The emergence and dissemination of vaccines against Streptococcus pneumoniae in disease prevention and control has a bright future.

  3. Evolution of antimicrobial resistance and serotype distribution of Streptococcus pneumoniae isolated from children with invasive and noninvasive pneumococcal diseases in Algeria from 2005 to 2012

    N. Ramdani-Bouguessa

    2015-07-01

    Full Text Available Pneumococcal infections are a major cause of morbidity and mortality in developing countries. The introduction of pneumococcal conjugate vaccines (PCVs has dramatically reduced the incidence of pneumococcal diseases. PCVs are not currently being used in Algeria. We conducted a prospective study from 2005 to 2012 in Algeria to determine antimicrobial drug resistance and serotype distribution of Streptococcus pneumoniae from children with pneumococcal disease. Among 270 isolated strains from children, 97 (36% were invasive disease; of these, 48% were not susceptible to penicillin and 53% not susceptible to erythromycin. A high rate of antimicrobial nonsusceptibility was observed in strains isolated from children with meningitis. The serotype distribution from pneumococci isolated from children with invasive infections was (by order of prevalence: 14, 1, 19F, 19A, 6B, 5, 3, 6A and 23F. Multidrug resistance was observed in serotypes 14, 19F, 19A and 6B. The vaccine coverage of serotypes isolated from children aged <5 years was 55.3% for PCV7, 71.1% for PCV10 and 86.8% for PCV13. Our results highlight the burden of pneumococcal disease in Algeria and the increasing S. pneumoniae antibiotic resistance. The current pneumococcal vaccines cover a high percentage of the circulating strains. Therefore, vaccination would reduce the incidence of pneumococcal disease in Algeria.

  4. Evolution of antimicrobial resistance and serotype distribution of Streptococcus pneumoniae isolated from children with invasive and noninvasive pneumococcal diseases in Algeria from 2005 to 2012.

    Ramdani-Bouguessa, N; Ziane, H; Bekhoucha, S; Guechi, Z; Azzam, A; Touati, D; Naim, M; Azrou, S; Hamidi, M; Mertani, A; Laraba, A; Annane, T; Kermani, S; Tazir, M

    2015-07-01

    Pneumococcal infections are a major cause of morbidity and mortality in developing countries. The introduction of pneumococcal conjugate vaccines (PCVs) has dramatically reduced the incidence of pneumococcal diseases. PCVs are not currently being used in Algeria. We conducted a prospective study from 2005 to 2012 in Algeria to determine antimicrobial drug resistance and serotype distribution of Streptococcus pneumoniae from children with pneumococcal disease. Among 270 isolated strains from children, 97 (36%) were invasive disease; of these, 48% were not susceptible to penicillin and 53% not susceptible to erythromycin. A high rate of antimicrobial nonsusceptibility was observed in strains isolated from children with meningitis. The serotype distribution from pneumococci isolated from children with invasive infections was (by order of prevalence): 14, 1, 19F, 19A, 6B, 5, 3, 6A and 23F. Multidrug resistance was observed in serotypes 14, 19F, 19A and 6B. The vaccine coverage of serotypes isolated from children aged <5 years was 55.3% for PCV7, 71.1% for PCV10 and 86.8% for PCV13. Our results highlight the burden of pneumococcal disease in Algeria and the increasing S. pneumoniae antibiotic resistance. The current pneumococcal vaccines cover a high percentage of the circulating strains. Therefore, vaccination would reduce the incidence of pneumococcal disease in Algeria. PMID:26106481

  5. M-ficolin binds selectively to the capsular polysaccharides of Streptococcus pneumoniae serotypes 19B and 19C and of a S. mitis strain

    Kjær, Troels Rønn; Hansen, Annette G; Sørensen, Uffe B S; Holm, Anne Trommelholt; Sørensen, Grith Lykke; Jensenius, Jens C; Thiel, Steffen

    2012-01-01

    infections. We investigated the binding selectivity by examining the binding of M-ficolin to a panel of more than 100 different streptococcal strains (Streptococcus pneumoniae and Streptococcus mitis) each expressing distinct polysaccharide structures. M-ficolin binding was observed for three strains only......-ficolin ligand.In conclusion, we were able to demonstrate specific binding of M-ficolin to some capsular polysaccharides of the opportunistic pathogen S. pneumoniae and of the commensal bacterium S. mitis....

  6. High-Level Genetic Diversity among Invasive Streptococcus pneumoniae Isolates in Turkey.

    Guldemir, Dilek; Acar, Sumeyra; Otgun, Selin Nar; Unaldi, Ozlem; Gozalan, Aysegul; Ertek, Mustafa; Durmaz, Riza

    2016-05-20

    This study obtained information on the serotypes and molecular typing characteristics of Streptococcus pneumoniae strains causing invasive diseases in Turkey. Sixty-eight S. pneumoniae isolates causing invasive pneumococcal diseases were collected from different regions of Turkey from 2009 to 2011. The isolates were characterized by performing multilocus sequence typing (MLST), pulsed-field gel electrophoresis (PFGE), and capsular serotyping, and 25 different serotypes were identified. Serotypes 19F, 23F, 1, 14, and 7F were common and accounted for 52.9% of all the serotypes. In addition, 54 different PFGE profiles (pulsotypes) were observed. Twenty-three of the 68 (33.8%) isolates were clustered into 9 pulsotypes. MLST analysis yielded 36 sequence types, of which 12 (33.3%) were novel. A comparison of results with the global pneumococcal MLST database by performing eBURST analysis showed that our strains belonged to 20 different clonal complexes and 5 singletons. In addition, we identified 4 new alleles: 2 gdh, 1 xpt, and 1 ddl. Thus, the results of this study highlighted a high level of diversity among pneumococcal isolates. In addition, the study identified a case of possible capsular switching. PMID:26255730

  7. Preparation of inocula for experimental infection of blood with Streptococcus pneumoniae

    Vivas-Alegre, Santiago; Fernández-Natal, Isabel; López-Fidalgo, Eduardo; Rivero-Lezcano, Octavio Miguel

    2015-01-01

    Experimental infections of either cells or animals require the preparation of good quality inocula. Unfortunately, the important pulmonary pathogen Streptococcus pneumoniae is a fastidious microorganism that suffers an autolysis process when cultured in vitro. Supplementation of Todd–Hewitt broth with a biological buffer (20 mM Tris–HCl, pH = 7.8) promotes a six hours delay in the beginning of the autolysis process. Additional improvements include washing bacteria before freezing, avoiding manipulations after thawing, and the use of glycerol (70% of the frozen bacteria was viable after 28 weeks at −80 °C, and aliquots were highly homogeneous. We have tested their utility in a whole blood infection model and have found that human plasma exhibits a higher microbicidal activity than whole blood, a result that we have not found previously reported. Additionally, we have also observed significant variations in the antimicrobial activity against different strains, which might be related to their virulence.•Media culture buffering extends S. pneumoniae viability for 6 h.•Washing before freezing of single use aliquots minimizes manipulation after thawing.•Experimental infection with the frozen inocula has shown that plasma has higher bactericidal activity than blood. PMID:26844211

  8. Transcriptional profiling of UlaR-regulated genes in Streptococcus pneumoniae

    Sulman Shafeeq

    2015-06-01

    Full Text Available The transcriptional regulator UlaR belongs to the family of PRD-containing transcriptional regulators, which are mostly involved in the regulation of carbohydrate metabolism. The role of the transcriptional regulator UlaR in Streptococcus pneumoniae has recently been described [1]. Here, we report detailed genome-wide transcriptional profiling of UlaR-regulated genes in S. pneumoniae D39 and its ∆ulaR derivative, either in the presence of 10 mM ascorbic acid in M17 medium using microarray analysis. 10 mM concentration of ascorbic acid was supplemented to the M17 medium because our lacZ-fusion studies indicated that UlaR acts as a transcriptional activator of its targets in the presence of ascorbic acid and the expression of the ula operon was maximal at a 10 mM ascorbic acid concentration [1]. All transcriptional profiling data of UlaR-regulated genes was deposited to Gene Expression Omnibus (GEO database under accession number GSE61649.

  9. Prophage spontaneous activation promotes DNA release enhancing biofilm formation in Streptococcus pneumoniae.

    Margarida Carrolo

    Full Text Available Streptococcus pneumoniae (pneumococcus is able to form biofilms in vivo and previous studies propose that pneumococcal biofilms play a relevant role both in colonization and infection. Additionally, pneumococci recovered from human infections are characterized by a high prevalence of lysogenic bacteriophages (phages residing quiescently in their host chromosome. We investigated a possible link between lysogeny and biofilm formation. Considering that extracellular DNA (eDNA is a key factor in the biofilm matrix, we reasoned that prophage spontaneous activation with the consequent bacterial host lysis could provide a source of eDNA, enhancing pneumococcal biofilm development. Monitoring biofilm growth of lysogenic and non-lysogenic pneumococcal strains indicated that phage-infected bacteria are more proficient at forming biofilms, that is their biofilms are characterized by a higher biomass and cell viability. The presence of phage particles throughout the lysogenic strains biofilm development implicated prophage spontaneous induction in this effect. Analysis of lysogens deficient for phage lysin and the bacterial major autolysin revealed that the absence of either lytic activity impaired biofilm development and the addition of DNA restored the ability of mutant strains to form robust biofilms. These findings establish that limited phage-mediated host lysis of a fraction of the bacterial population, due to spontaneous phage induction, constitutes an important source of eDNA for the S. pneumoniae biofilm matrix and that this localized release of eDNA favors biofilm formation by the remaining bacterial population.

  10. Antimicrobial Activity of Novel Synthetic Peptides Derived from Indolicidin and Ranalexin against Streptococcus pneumoniae.

    Hassan Mahmood Jindal

    Full Text Available Antimicrobial peptides (AMPs represent promising alternatives to conventional antibiotics in order to defeat multidrug-resistant bacteria such as Streptococcus pneumoniae. In this study, thirteen antimicrobial peptides were designed based on two natural peptides indolicidin and ranalexin. Our results revealed that four hybrid peptides RN7-IN10, RN7-IN9, RN7-IN8, and RN7-IN6 possess potent antibacterial activity against 30 pneumococcal clinical isolates (MIC 7.81-15.62µg/ml. These four hybrid peptides also showed broad spectrum antibacterial activity (7.81µg/ml against S. aureus, methicillin resistant S. aureus (MRSA, and E. coli. Furthermore, the time killing assay results showed that the hybrid peptides were able to eliminate S. pneumoniae within less than one hour which is faster than the standard drugs erythromycin and ceftriaxone. The cytotoxic effects of peptides were tested against human erythrocytes, WRL-68 normal liver cell line, and NL-20 normal lung cell line. The results revealed that none of the thirteen peptides have cytotoxic or hemolytic effects at their MIC values. The in silico molecular docking study was carried out to investigate the binding properties of peptides with three pneumococcal virulent targets by Autodock Vina. RN7IN6 showed a strong affinity to target proteins; autolysin, pneumolysin, and pneumococcal surface protein A (PspA based on rigid docking studies. Our results suggest that the hybrid peptides could be suitable candidates for antibacterial drug development.

  11. Overexpression, purification and crystallization of a choline-binding protein CbpI from Streptococcus pneumoniae

    Paterson, Neil G., E-mail: neison@chem.gla.ac.uk; Riboldi-Tunicliffe, Alan [Department of Chemistry and WestCHEM, Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom); Mitchell, Timothy J. [Division of Infection and Immunity (IBLS), Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom); Isaacs, Neil W. [Department of Chemistry and WestCHEM, Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom)

    2006-07-01

    The choline-binding protein CbpI from S. pneumoniae has been purified and crystallized and diffraction data have been collected to 3.5 Å resolution. The choline-binding protein CbpI from Streptococcus pneumoniae is a 23.4 kDa protein with no known function. The protein has been successfully purified initially using Ni–NTA chromatography and to homogeneity using Q-Sepharose ion-exchange resin as an affinity column. CbpI was crystallized using PEG 3350 as a precipitant and X-ray crystallographic analysis showed that the crystals belonged to the tetragonal space group P4, with unit-cell parameters a = b = 83.31, c = 80.29 Å, α = β = γ = 90°. The crystal contains two molecules in the asymmetric unit with a solvent content of 55.7% (V{sub M} = 2.77 Å{sup 3} Da{sup −1}) and shows a diffraction limit of 3.5 Å.

  12. Discovery of prenylated flavonoids with dual activity against influenza virus and Streptococcus pneumoniae

    Grienke, Ulrike; Richter, Martina; Walther, Elisabeth; Hoffmann, Anja; Kirchmair, Johannes; Makarov, Vadim; Nietzsche, Sandor; Schmidtke, Michaela; Rollinger, Judith M.

    2016-01-01

    Influenza virus neuraminidase (NA) is the primary target for influenza therapeutics. Severe complications are often related to secondary pneumonia caused by Streptococcus pneumoniae (pneumococci), which also express NAs. Recently, a NA-mediated lethal synergism between influenza A viruses and pneumococci was described. Therefore, dual inhibitors of both viral and bacterial NAs are expected to be advantageous for the treatment of influenza. We investigated the traditional Chinese herbal drug sāng bái pí (mulberry root bark) as source for anti-infectives. Two prenylated flavonoid derivatives, sanggenon G (4) and sanggenol A (5) inhibited influenza A viral and pneumococcal NAs and, in contrast to the approved NA inhibitor oseltamivir, also planktonic growth and biofilm formation of pneumococci. Evaluation of 27 congeners of 5 revealed a correlation between the degree of prenylation and bioactivity. Abyssinone-V 4′-methyl ether (27) inhibited pneumococcal NA with IC50 = 2.18 μM, pneumococcal growth with MIC = 5.63 μM, and biofilm formation with MBIC = 4.21 μM, without harming lung epithelial cells. Compounds 5 and 27 also disrupt the synergism between influenza A virus and pneumococcal NA in vitro, hence functioning as dual-acting anti-infectives. The results warrant further studies on whether the observed disruption of this synergism is transferable to in vivo systems. PMID:27257160

  13. Adenylate kinase from Streptococcus pneumoniae is essential for growth through its catalytic activity

    Trung Thanh Thach

    2014-01-01

    Full Text Available Streptococcus pneumoniae (pneumococcus infection causes more than 1.6 million deaths worldwide. Pneumococcal growth is a prerequisite for its virulence and requires an appropriate supply of cellular energy. Adenylate kinases constitute a major family of enzymes that regulate cellular ATP levels. Some bacterial adenylate kinases (AdKs are known to be critical for growth, but the physiological effects of AdKs in pneumococci have been poorly understood at the molecular level. Here, by crystallographic and functional studies, we report that the catalytic activity of adenylate kinase from S. pneumoniae (SpAdK serotype 2 D39 is essential for growth. We determined the crystal structure of SpAdK in two conformations: ligand-free open form and closed in complex with a two-substrate mimic inhibitor adenosine pentaphosphate (Ap5A. Crystallographic analysis of SpAdK reveals Arg-89 as a key active site residue. We generated a conditional expression mutant of pneumococcus in which the expression of the adk gene is tightly regulated by fucose. The expression level of adk correlates with growth rate. Expression of the wild-type adk gene in fucose-inducible strains rescued a growth defect, but expression of the Arg-89 mutation did not. SpAdK increased total cellular ATP levels. Furthermore, lack of functional SpAdK caused a growth defect in vivo. Taken together, our results demonstrate that SpAdK is essential for pneumococcal growth in vitro and in vivo.

  14. Cirrhosis-induced defects in innate pulmonary defenses against Streptococcus pneumoniae

    Vander Top Elizabeth A

    2007-10-01

    Full Text Available Abstract Background The risk of mortality from pneumonia caused by Streptococcus pneumoniae is increased in patients with cirrhosis. However, the specific pneumococcal virulence factors and host immune defects responsible for this finding have not been clearly established. This study used a cirrhotic rat model of pneumococcal pneumonia to identify defect(s in innate pulmonary defenses in the cirrhotic host and to determine the impact of the pneumococcal toxin pneumolysin on these defenses in the setting of severe cirrhosis. Results No cirrhosis-associated defects in mucociliary clearance of pneumococci were found in these studies, but early intrapulmonary killing of the organisms before the arrival of neutrophils was significantly impaired. This defect was exacerbated by pneumolysin production in cirrhotic but not in control rats. Neutrophil-mediated killing of a particularly virulent type 3 pneumococcal strain also was significantly diminished within the lungs of cirrhotic rats with ascites. Levels of lysozyme and complement component C3 were both significantly reduced in bronchoalveolar lavage fluid from cirrhotic rats. Finally, complement deposition was reduced on the surface of pneumococci recovered from the lungs of cirrhotic rats in comparison to organisms recovered from the lungs of control animals. Conclusion Increased mortality from pneumococcal pneumonia in this cirrhotic host is related to defects in both early pre-neutrophil- and later neutrophil-mediated pulmonary killing of the organisms. The fact that pneumolysin production impaired pre-neutrophil-mediated pneumococcal killing in cirrhotic but not control rats suggests that pneumolysin may be particularly detrimental to this defense mechanism in the severely cirrhotic host. The decrease in neutrophil-mediated killing of pneumococci within the lungs of the cirrhotic host is related to insufficient deposition of host proteins such as complement C3 on their surfaces. Pneumolysin

  15. Phenotypic and genotypic characterization of Streptococcus pneumoniae resistant to macrolide in Casablanca, Morocco.

    Diawara, Idrissa; Zerouali, Khalid; Katfy, Khalid; Barguigua, Abouddihaj; Belabbes, Houria; Timinouni, Mohammed; Elmdaghri, Naima

    2016-06-01

    In Morocco, the 13-valent pneumococcal conjugate vaccine (PCV-13) was introduced in the national immunization program (NIP) in October 2010 and replaced by the PCV-10 in July 2012. The present study aimed to determine the prevalence of erythromycin-resistant Streptococcus pneumoniae (ERSP) and to analyze the phenotypic and genotypic characteristics of these isolates in Casablanca, Morocco from January 2007 to December 2014. Isolates were obtained from the Microbiology Laboratory of Ibn Rochd University Hospital Centre of Casablanca. Serogrouping was done using Pneumotest Kit and serotyping by the Quellung capsular swelling. Antibiotic susceptibility pattern was determined by disk diffusion and Etest methods. A total of 655S. pneumoniae isolates were collected from 2007 to 2014 from pediatric and adult patients. Fifty-five percent of these isolates were from invasive pneumococcal diseases. Of the 655 isolates, 92 (14%) were ERSP. Globally, the proportion of ERSP from 2007 to 2010 (before vaccination) and from 2011 to 2014 (after vaccination) were 11.6% and 17.2% (p=0.04), respectively. Of the 92 ERSP, 89%, 4% and 7% displayed constitutive MLSB (resistance to macrolide, lincosamide and streptogramin B), inducible MLSB, and M phenotype (resistance to macrolide only), respectively. ERSP genotypic analysis showed that 90.2% carried the ermB gene, 6.5% the mefE gene, and 3.3% both the genes (ermB+mefE). The most prevalent ERSP serotypes were 6B, 19F and 23F before vaccination and 19F, 6B, 6A and 23F after vaccination. Erythromycin resistance among S. pneumoniae is relatively high in Casablanca. The contribution of PCVs to the reduction in antibiotic use is encouraging but this should be accompanied by a rational use of antibiotic. PMID:26961592

  16. A new variant of the capsule 3 cluster occurs in Streptococcus pneumoniae from deceased wild chimpanzees.

    Denapaite, Dalia; Hakenbeck, Regine

    2011-01-01

    The presence of new Streptococcus pneumoniae clones in dead wild chimpanzees from the Taï National Park, Côte d'Ivoire, with previous respiratory problems has been demonstrated recently by DNA sequence analysis from samples obtained from the deceased apes. In order to broadenour understanding on the relatedness of these pneumococcal clones to those from humans, the gene locus responsible for biosynthesis of the capsule polysaccharide (CPS) has now been characterized. DNA sequence analysis of PCR fragments identified a cluster named cps3(Taï) containing the four genes typical for serotype 3 CPS, but lacking a 5'-region of ≥2 kb which is degenerated in other cps3 loci and not required for type 3 biosynthesis. CPS3 is composed of a simple disaccharide repeat unit comprising glucose and glucuronic acid (GlcUA). The two genes ugd responsible for GlcUA synthesis and wchE encoding the type 3 synthase are essential for CPS3 biosynthesis, whereas both, galU and the 3'-truncated gene pgm are not required due to the presence of homologues elsewhere in the genome. The DNA sequence of cps3(Taï) diverged considerably from those of other cps3 loci. Also, the gene pgm(Taï) represents a full length version with a nonsense mutation at codon 179. The two genes ugd(Taï) and wchE(Taï) including the promoter region were transformed into a nonencapsulated laboratory strain S. pneumoniae R6. Transformants which expressed type 3 capsule polysaccharide were readily obtained, documenting that the gene products are functional. In summary, the data indicate that cps3(Taï) evolved independent from other cps3 loci, suggesting the presence of specialized serotype 3 S. pneumoniae clones endemic to the Taï National Park area. PMID:21969869

  17. Exposure to welding fumes and lower airway infection with Streptococcus pneumoniae

    Suri, Reetika; Periselneris, Jimstan; Lanone, Sophie; Zeidler-Erdely, Patti C.; Melton, Geoffrey; Palmer, Keith T.; Andujar, Pascal; Antonini, James M.; Cohignac, Vanessa; Erdely, Aaron; Jose, Ricardo J.; Mudway, Ian; Brown, Jeremy; Grigg, Jonathan

    2015-01-01

    Background Welders are at increased risk of pneumococcal pneumonia. The mechanism for this association is not known. The capacity of pneumococci to adhere to and infect lower airway cells is mediated by host-expressed platelet-activating factor receptor (PAFR). Objective We sought to assess the effect of mild steel welding fumes (MS-WF) on PAFR-dependent pneumococcal adhesion and infection to human airway cells in vitro and on pneumococcal airway infection in a mouse model. Methods The oxidative potential of MS-WF was assessed by their capacity to reduce antioxidants in vitro. Pneumococcal adhesion and infection of A549, BEAS-2B, and primary human bronchial airway cells were assessed by means of quantitative bacterial culture and expressed as colony-forming units (CFU). After intranasal instillation of MS-WF, mice were infected with Streptococcus pneumoniae, and bronchoalveolar lavage fluid (BALF) and lung CFU values were determined. PAFR protein levels were assessed by using immunofluorescence and immunohistochemistry, and PAFR mRNA expression was assessed by using quantitative PCR. PAFR was blocked by CV-3988, and oxidative stress was attenuated by N-acetylcysteine. Results: MS-WF exhibited high oxidative potential. In A549 and BEAS-2B cells MS-WF increased pneumococcal adhesion and infection and PAFR protein expression. Both CV-3988 and N-acetylcysteine reduced MS-WF–stimulated pneumococcal adhesion and infection of airway cells. MS-WF increased mouse lung PAFR mRNA expression and increased BALF and lung pneumococcal CFU values. In MS-WF–exposed mice CV-3988 reduced BALF CFU values. Conclusions Hypersusceptibility of welders to pneumococcal pneumonia is in part mediated by the capacity of welding fumes to increase PAFR-dependent pneumococcal adhesion and infection of lower airway cells. PMID:26277596

  18. A new variant of the capsule 3 cluster occurs in Streptococcus pneumoniae from deceased wild chimpanzees.

    Dalia Denapaite

    Full Text Available The presence of new Streptococcus pneumoniae clones in dead wild chimpanzees from the Taï National Park, Côte d'Ivoire, with previous respiratory problems has been demonstrated recently by DNA sequence analysis from samples obtained from the deceased apes. In order to broadenour understanding on the relatedness of these pneumococcal clones to those from humans, the gene locus responsible for biosynthesis of the capsule polysaccharide (CPS has now been characterized. DNA sequence analysis of PCR fragments identified a cluster named cps3(Taï containing the four genes typical for serotype 3 CPS, but lacking a 5'-region of ≥2 kb which is degenerated in other cps3 loci and not required for type 3 biosynthesis. CPS3 is composed of a simple disaccharide repeat unit comprising glucose and glucuronic acid (GlcUA. The two genes ugd responsible for GlcUA synthesis and wchE encoding the type 3 synthase are essential for CPS3 biosynthesis, whereas both, galU and the 3'-truncated gene pgm are not required due to the presence of homologues elsewhere in the genome. The DNA sequence of cps3(Taï diverged considerably from those of other cps3 loci. Also, the gene pgm(Taï represents a full length version with a nonsense mutation at codon 179. The two genes ugd(Taï and wchE(Taï including the promoter region were transformed into a nonencapsulated laboratory strain S. pneumoniae R6. Transformants which expressed type 3 capsule polysaccharide were readily obtained, documenting that the gene products are functional. In summary, the data indicate that cps3(Taï evolved independent from other cps3 loci, suggesting the presence of specialized serotype 3 S. pneumoniae clones endemic to the Taï National Park area.

  19. Streptococcus pneumoniae Enhances Human Respiratory Syncytial Virus Infection In Vitro and In Vivo.

    D Tien Nguyen

    Full Text Available Human respiratory syncytial virus (HRSV and Streptococcus pneumoniae are important causative agents of respiratory tract infections. Both pathogens are associated with seasonal disease outbreaks in the pediatric population, and can often be detected simultaneously in infants hospitalized with bronchiolitis or pneumonia. It has been described that respiratory virus infections may predispose for bacterial superinfections, resulting in severe disease. However, studies on the influence of bacterial colonization of the upper respiratory tract on the pathogenesis of subsequent respiratory virus infections are scarce. Here, we have investigated whether pneumococcal colonization enhances subsequent HRSV infection. We used a newly generated recombinant subgroup B HRSV strain that expresses enhanced green fluorescent protein and pneumococcal isolates obtained from healthy children in disease-relevant in vitro and in vivo model systems. Three pneumococcal strains specifically enhanced in vitro HRSV infection of primary well-differentiated normal human bronchial epithelial cells grown at air-liquid interface, whereas two other strains did not. Since previous studies reported that bacterial neuraminidase enhanced HRSV infection in vitro, we measured pneumococcal neuraminidase activity in these cultures but found no correlation with the observed infection enhancement in our model. Subsequently, a selection of pneumococcal strains was used to induce nasal colonization of cotton rats, the best available small animal model for HRSV. Intranasal HRSV infection three days later resulted in strain-specific enhancement of HRSV replication in vivo. One S. pneumoniae strain enhanced HRSV both in vitro and in vivo, and was also associated with enhanced syncytium formation in vivo. However, neither pneumococci nor HRSV were found to spread from the upper to the lower respiratory tract, and neither pathogen was transmitted to naive cage mates by direct contact. These

  20. Extracellular Adenosine Protects against Streptococcus pneumoniae Lung Infection by Regulating Pulmonary Neutrophil Recruitment.

    Bou Ghanem, Elsa N; Clark, Stacie; Roggensack, Sara E; McIver, Sally R; Alcaide, Pilar; Haydon, Philip G; Leong, John M

    2015-08-01

    An important determinant of disease following Streptococcus pneumoniae (pneumococcus) lung infection is pulmonary inflammation mediated by polymorphonuclear leukocytes (PMNs). We found that upon intratracheal challenge of mice, recruitment of PMNs into the lungs within the first 3 hours coincided with decreased pulmonary pneumococci, whereas large numbers of pulmonary PMNs beyond 12 hours correlated with a greater bacterial burden. Indeed, mice that survived infection largely resolved inflammation by 72 hours, and PMN depletion at peak infiltration, i.e. 18 hours post-infection, lowered bacterial numbers and enhanced survival. We investigated host signaling pathways that influence both pneumococcus clearance and pulmonary inflammation. Pharmacologic inhibition and/or genetic ablation of enzymes that generate extracellular adenosine (EAD) (e.g. the ectoenzyme CD73) or degrade EAD (e.g. adenosine deaminase) revealed that EAD dramatically increases murine resistance to S. pneumoniae lung infection. Moreover, adenosine diminished PMN movement across endothelial monolayers in vitro, and although inhibition or deficiency of CD73 had no discernible impact on PMN recruitment within the first 6 hours after intratracheal inoculation of mice, these measures enhanced PMN numbers in the pulmonary interstitium after 18 hours of infection, culminating in dramatically elevated numbers of pulmonary PMNs at three days post-infection. When assessed at this time point, CD73-/- mice displayed increased levels of cellular factors that promote leukocyte migration, such as CXCL2 chemokine in the murine lung, as well as CXCR2 and β-2 integrin on the surface of pulmonary PMNs. The enhanced pneumococcal susceptibility of CD73-/- mice was significantly reversed by PMN depletion following infection, suggesting that EAD-mediated resistance is largely mediated by its effects on PMNs. Finally, CD73-inhibition diminished the ability of PMNs to kill pneumococci in vitro, suggesting that EAD alters

  1. Contribución al estudio de los mecanismos moleculares de la resistencia a ciprofloxacina en "Streptococcus pneumoniae"

    Martínez Garriga, Blanca

    2005-01-01

    TESIS DOCTORAL"Streptococcus pneumoniae", también llamado neumococo, es una bacteria Gram positiva caracterizada por la severidad de las infecciones que produce en la especie humana, siendo responsable de una elevada morbilidad y letalidad, especialmente en niños y ancianos, colectivos particularmente sensibles a sus consecuencias. La emergencia de cepas de S. pneumoniae resistentes a antibióticos como la penicilina y otros beta-lactámicos, comúnmente utilizados en el tratamiento de las infec...

  2. Surveillance of bacterial resistance in Streptococcus pneumoniae%肺炎链球菌的耐药性监测

    张泓

    2011-01-01

    As the prevalence of resistant Streptococcus pneumoniae has been increased significantly, understanding the characteristic of resistant strains and applying surveillance for them are helpful to guide clinical therapy and control the prevalence of resistant strains. This review describes Streptococcus pneumoniae resistance, resistance mechanism and their clinical surveillance.%肺炎链球菌耐药率不断上升,监测其耐药性并掌握耐药特征,有助于指导临床合理选药及控制肺炎链球菌耐药株流行.本文综述肺炎链球菌的耐药现状、相关机制及监测方法.

  3. Toll-Like Receptor Signalling Is Not Involved in Platelet Response to Streptococcus pneumoniae In Vitro or In Vivo

    Schaap, Marianne C. L.; Hou, Baidong; van der Poll, Tom; Nieuwland, Rienk; van ‘t Veer, Cornelis

    2016-01-01

    Streptococcus (S.) pneumoniae strains vary considerably in their ability to cause invasive disease in humans, which is at least in part determined by the capsular serotype. Platelets have been implicated as sentinel cells in the circulation for host defence. One of their utensils for this function is the expression of Toll-like receptors (TLRs). We here aimed to investigate platelet response to S. pneumoniae and a role for TLRs herein. Platelets were stimulated using four serotypes of S. pneumonia including an unencapsulated mutant strain. In vitro aggregation and flow cytometry assays were performed using blood of healthy volunteers, or blood of TLR knock out and WT mice. For in vivo pneumonia experiments, platelet specific Myd88 knockout (Plt-Myd88-/-) mice were used. We found that platelet aggregation was induced by unencapsulated S. pneumoniae only. Whole blood incubation with all S. pneumoniae serotypes tested resulted in platelet degranulation and platelet-leukocyte complex formation. Platelet activation was TLR independent, as responses were not inhibited by TLR blocking antibodies, not induced by TLR agonists and were equally induced in wild-type and Tlr2-/-, Tlr4-/-, Tlr2/4-/-, Tlr9-/- and Myd88-/- blood. Plt-Myd88-/- and control mice displayed no differences in bacterial clearance or immune response to pneumonia by unencapsulated S. pneumoniae. In conclusion, S. pneumoniae activates platelets through a TLR-independent mechanism that is impeded by the bacterial capsule. Additionally, platelet MyD88-dependent TLR signalling is not involved in host defence to unencapsulated S. pneumoniae in vivo. PMID:27253707

  4. Telithromycin resistance in Streptococcus pneumoniae is conferred by a deletion in the leader sequence of erm(B) that increases rRNA methylation

    Wolter, Nicole; Smith, Anthony M; Farrell, David J; Northwood, John Blackman; Douthwaite, Stephen; Klugman, Keith P

    2008-01-01

    A telithromycin-resistant clinical isolate of Streptococcus pneumoniae (strain P1501016) has been found to contain a version of erm(B) that is altered by a 136-bp deletion in the leader sequence. By allele replacement mutagenesis, a second strain of S. pneumoniae (PC13) with a wild-type erm(B) gene...

  5. Phenotypic and Genotypic Characterization of Streptococcus pneumoniae Strains Colonizing Children Attending Day-Care Centers in Norway▿

    Vestrheim, Didrik F.; Høiby, E. Arne; Aaberge, Ingeborg S; Dominique A. Caugant

    2008-01-01

    A cross-sectional study of nasopharyngeal colonization with Streptococcus pneumoniae was performed among 573 children attending 29 day-care centers (DCCs) in Norway prior to the start of mass vaccination with the heptavalent pneumococcal conjugate vaccine (PCV-7). A sensitive sampling method was employed, including transport in an enrichment broth and serotyping of pneumococci directly from the broth, in addition to traditional single-colony isolation from blood agar plates. The prevalence of...

  6. Cloning and nucleotide sequence of the Streptococcus pneumoniae hyaluronidase gene and purification of the enzyme from recombinant Escherichia coli.

    Berry, A M; Lock, R A; Thomas, S M; Rajan, D P; Hansman, D.; Paton, J C

    1994-01-01

    A gene bank of Sau3A1-generated Streptococcus pneumoniae type 23 DNA fragments was constructed in Escherichia coli K-12 with the low-copy-number cosmid vector pOU61cos. Clone lysates were screened by immunoblotting using a mouse antiserum raised against a crude pneumococcal hyaluronidase preparation. One immunoreactive clone was isolated, and it produced high level of hyaluronidase activity. This clone contained a recombinant cosmid (designated pJCP800) with an approximately 35-kb DNA insert,...

  7. beta-lactamase-producing nontypeable Haemophilus influenzae fails to protect Streptococcus pneumoniae from amoxicillin during experimental acute otitis media

    Westman, E.; Lundin, S.; Hermansson, Ann; Melhus, Åsa

    2004-01-01

    Acute otitis media (AOM) is the most common reason for outpatient antimicrobial therapy. Mixed infections pose a potential problem, since the first-line drug used for the treatment of AOM, amoxicillin, can be neutralized by ß-lactamase-producing pathogens of the upper respiratory tract. To study the effects of a 5-day course of amoxicillin on a mixed middle ear infection, rats were challenged with Streptococcus pneumoniae alone or in combination with ß-lactamase-producing nontypeable Haemophi...

  8. β-Lactamase-Producing Nontypeable Haemophilus influenzae Fails To Protect Streptococcus pneumoniae from Amoxicillin during Experimental Acute Otitis Media

    Westman, Eva; Lundin, Susanne; Hermansson, Ann; Melhus, Åsa

    2004-01-01

    Acute otitis media (AOM) is the most common reason for outpatient antimicrobial therapy. Mixed infections pose a potential problem, since the first-line drug used for the treatment of AOM, amoxicillin, can be neutralized by β-lactamase-producing pathogens of the upper respiratory tract. To study the effects of a 5-day course of amoxicillin on a mixed middle ear infection, rats were challenged with Streptococcus pneumoniae alone or in combination with β-lactamase-producing nontypeable Haemophi...

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

    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...

  10. Eradication of Streptococcus pneumoniae in the Nasopharyngeal Flora of Children with Acute Otitis Media after Amoxicillin-Clavulanate Therapy

    Brook, Itzhak; Gober, Alan E.

    2004-01-01

    Nasopharyngeal cultures were obtained from 60 children with acute otitis media before and after treatment with either 45 or 90 mg of amoxicillin (given as amoxicillin-clavulanate) per kg of body weight per day for 10 days. The number of Streptococcus pneumoniae isolates in the 45-mg/kg group was reduced from 12 to 6 and was reduced from 14 to 1 (P = 0.0261) in the 90-mg/kg group.

  11. Activity of Amoxicillin-Clavulanate against Penicillin-Resistant Streptococcus pneumoniae in an Experimental Respiratory Infection Model in Rats†

    Smith, Gillian M.; Slocombe, Brian; Abbott, Karen H.; Mizen, Linda W.

    1998-01-01

    High doses of amoxicillin, equivalent to those produced by 500- and 750-mg oral doses in humans (area under the plasma concentration-time curve), were effective against a penicillin-resistant strain of Streptococcus pneumoniae in an experimental respiratory tract infection in immunocompromised rats; this superior activity confirms the results of previous studies. An unexpected enhancement of amoxicillin’s antibacterial activity in vivo against penicillin-resistant and -susceptible S. pneumoni...

  12. Comparison of different culture media and storage temperatures for the long-term preservation of Streptococcus pneumoniae in the tropics.

    2001-01-01

    OBJECTIVE: The preservation of Streptococcus pneumoniae by standard freezing methods for subsequent tests--such as serotyping and antibiotic susceptibility--is not possible or is difficult in many developing countries because of the high cost of equipment, inadequate equipment maintenance, and irregular power supply. We evaluated alternative low-cost methods, by comparing different culture media and storage temperatures. METHODS: Clinical isolates of five capsular types (1, 5, 7, 19, and 23) ...

  13. In Vitro Activities of Telithromycin, Linezolid, and Quinupristin-Dalfopristin against Streptococcus pneumoniae with Macrolide Resistance Due to Ribosomal Mutations

    Farrell, David J.; Morrissey, Ian; Bakker, Sarah; Buckridge, Sylvie; Felmingham, David

    2004-01-01

    To date, 86 of 7,746 macrolide-resistant Streptococcus pneumoniae isolates from 1999 to 2002 PROTEKT (Prospective Resistant Organism Tracking and Epidemiology for the Ketolide Telithromycin) surveillance studies were negative for methylase and efflux mechanisms. Mutations in 23S rRNA or the genes encoding riboprotein L4 or L22 were found in 77 of 86 isolates. Six isolates were resistant to quinupristin-dalfopristin and two were resistant to linezolid, while telithromycin demonstrated good act...

  14. Development of Doxycycline MIC and Disk Diffusion Interpretive Breakpoints and Revision of Tetracycline Breakpoints for Streptococcus pneumoniae

    Dallas, Steven D.; McGee, Lesley; Limbago, Brandi; Patel, Jean B; McElmeel, M. Leticia; Fulcher, Letitia C.; Lonsway, David R.; Jorgensen, James H.

    2013-01-01

    A study was performed to derive susceptibility testing interpretive breakpoints for doxycycline with Streptococcus pneumoniae and to reassess breakpoints for tetracycline using the requirements defined in Clinical and Laboratory Standards Institute (CLSI) document M23-A3. Tetracycline and doxycycline MICs and disk diffusion zone sizes were determined on 189 isolates selected from the 2009-2010 CDC Active Bacterial Core surveillance strain collection according to the testing methods described ...

  15. TRAIL+ monocytes and monocyte-related cells cause lung damage and thereby increase susceptibility to influenza–Streptococcus pneumoniae coinfection

    Ellis, Gregory T; Davidson, Sophia; Crotta, Stefania; Branzk, Nora; Papayannopoulos, Venizelos; Wack, Andreas

    2015-01-01

    Streptococcus pneumoniae coinfection is a major cause of influenza-associated mortality; however, the mechanisms underlying pathogenesis or protection remain unclear. Using a clinically relevant mouse model, we identify immune-mediated damage early during coinfection as a new mechanism causing susceptibility. Coinfected CCR2−/− mice lacking monocytes and monocyte-derived cells control bacterial invasion better, show reduced epithelial damage and are overall more resistant than wild-type contr...

  16. Dimerization of Streptococcus pneumoniae eukaryotic-type serine/threonine protein kinase is a prerequisite of its catalytic activity

    Pallová, Petra; Přenosilová, Lenka; Nováková, Linda; Branny, Pavel

    Saint Malo : Verlag, 2006, s. 74-74. [ASM Conferences Streptococcal Genetics. Saint Malo (FR), 18.06.2006-21.06.2006] Grant ostatní: GA UK(CZ) 188/2004/B-BIO/PrF to LS; GA UK(CZ) 153/2006/B-BIO/PrF to PP Institutional research plan: CEZ:AV0Z50200510 Keywords : streptococcus pneumoniae * protein kinase Subject RIV: EE - Microbiology, Virology

  17. Rabbit antibodies to the cell wall polysaccharide of Streptococcus pneumoniae fail to protect mice from lethal challenge with encapsulated pneumococci.

    Szu, S C; Schneerson, R; Robbins, J B

    1986-01-01

    A conjugate, composed of the cell wall polysaccharide (C polysaccharide) of Streptococcus pneumoniae and bovine serum albumin (BSA), was prepared with the bifunctional agent N-succinimidyl-3-(2-pyridyldithio)-propionate. Analysis with monoclonal antibodies provided evidence that the phosphocholine (PC) moiety of the C polysaccharide was retained during the conjugation procedure. The C polysaccharide-BSA conjugate elicited antibodies to C polysaccharide in rabbits; no PC-specific antibodies we...

  18. Streptococcus pneumoniae Infection of Host Epithelial Cells via Polymeric Immunoglobulin Receptor Transiently Induces Calcium Release from Intracellular Stores*

    Asmat, T. M.; Agarwal, V; Rath, S.; Hildebrandt, J.-P.; Hammerschmidt, S.

    2011-01-01

    The pneumococcal surface protein C (PspC) is a major adhesin of Streptococcus pneumoniae (pneumococci) that interacts in a human-specific manner with the ectodomain of the human polymeric immunoglobulin receptor (pIgR) produced by respiratory epithelial cells. This interaction promotes bacterial colonization and bacterial internalization by initiating host signal transduction cascades. Here, we examined alterations of intracellular calcium ([Ca2+]i) levels in epithelial cells during host cell...

  19. Influence of clavulanic acid on the activity of amoxicillin against an experimental Streptococcus pneumoniae-Staphylococcus aureus mixed respiratory infection.

    Smith, G.M; Boon, R J; Beale, A S

    1990-01-01

    An experimental respiratory infection caused by Streptococcus pneumoniae was established in weanling rats by intrabronchial instillation. Treatment of this infection with amoxicillin rapidly eliminated the pneumococci from the lung tissue. A beta-lactamase-producing strain of Staphylococcus aureus, when inoculated in a similar manner, did not persist adequately in the lungs long enough to permit a reasonable assessment of the therapy, but staphylococcal survival was extended in the lungs of r...

  20. Genetics of oxacillin resistance in clinical isolates of Streptococcus pneumoniae that are oxacillin resistant and penicillin susceptible.

    Dowson, C G; Johnson, A P; Cercenado, E.; George, R C

    1994-01-01

    It has recently been reported that penicillin-sensitive pneumococci may exhibit reduced susceptibility to oxacillin, resulting in their misclassification as being penicillin resistant by oxacillin disk testing. Intermediate oxacillin resistance (MIC, 1.0 microgram/ml) in three of these apparently unrelated penicillin-susceptible clinical isolates of Streptococcus pneumoniae isolated in the United Kingdom and in four Spanish isolates was shown to be solely due to the acquisition of a gene enco...

  1. EFFECTS OF MICROWAVE-EXPOSURE AND TEMPERATURE ON SURVIVAL OF MICE INFECTED WITH 'STREPTOCOCCUS PNEUMONIAE' (JOURNAL VERSION)

    Female CD-1 mice were injected with an LD50 dose of Streptococcus pneumoniae and then exposed to 2.45 GHz (CW) microwave radiation at an incident power density of 10 mW/cm2 (SAR approximately equals 6.8 W/kg), 4 h/d for 5 d at ambient temperatures of 19, 22, 25, 28, 31, 34, 37 an...

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

    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

  3. Nasopharyngeal carriage, antibiogram & serotype distribution of Streptococcus pneumoniae among healthy under five children

    K.L. Ravi Kumar

    2014-01-01

    Full Text Available Background & objectives: Information related to nasopharyngeal carriage of Streptococcus pneumoniae among healthy children is scanty in India. This prospective study was undertaken to determine the presence of asymptomatic nasopharyngeal colonization, assess serogroups/types (SGT and drug resistance of S. pneumoniae in children below five years of age. Methods: A total of 109 male and 81 female children in the age group of three months to five years belonging to different socio-economic classes were enrolled. They were recruited across all age groups from those attending paediatric OPD of a tertiary care and research centre for immunization program. Fifty three isolates identified as pneumococci were tested for their antimicrobial susceptibility pattern by Kirby-Bauer′s disc diffusion and E-Test methods. Serotyping was performed by detection of the quelling reaction with specific antiserum. Result: The pneumococcal carriage rate in the study population was 27.9 per cent. The isolation rate was associated with age being higher (49.2% in smaller children (3-12 months and among male (62.2%. The most prevalent SGTs were 19 followed by 10, 14 and 7; 21 per cent of isolates belonging to serotype 10 (n=7 were 11 (n=4 were not covered in any of the conjugate vaccines currently available in Indian market. Resistance to co-trimoxazole, tetracycline, penicillin and erythromycin was observed in 91 per cent (n=48, 36 per cent (n=19, 17 per cent (n=9 and 9 per cent (n=5 isolates, respectively. All the penicillin resistant isolates were found to be intermediately resistant by E-Test. Multidrug resistance was observed in 19 per cent (n=10 isolates. Interpretation & conclusions: High level of antibiotic resistance was present in S. pneumoniae isolated from healthy children below age five. A pneumococcal conjugate vaccine with the prevailing SGTs would help to reduce the pool of antibiotic resistant pneumococci. Continued surveillance of serotypes and tracking

  4. Post-infective transverse myelitis following Streptococcus pneumoniae meningitis with radiological features of acute disseminated encephalomyelitis: a case report

    Williams Thomas

    2012-09-01

    Full Text Available Abstract Introduction Post-infectious autoimmune demyelination of the central nervous system is a rare neurological disorder typically associated with exanthematous viral infections. We report an unusual presentation of the condition and a previously undocumented association with Streptococcus pneumonia meningitis. Case presentation A 50-year-old Caucasian woman presented to our facility with an acute myelopathy three days after discharge following acute Streptococcus pneumoniae meningitis. Imaging studies of the spine ruled out an infective focus and no other lesions were seen within the cord. Diffuse, bilateral white matter lesions were seen within the cerebral hemispheres, and our patient was diagnosed as having a post-infective demyelination syndrome that met the diagnostic criteria for an acute transverse myelitis. Our patient clinically and radiologically improved following treatment with steroids. Conclusions The novel association of a Streptococcus pneumoniae infection with post-infectious autoimmune central nervous system demyelination should alert the reader to the potentially causative role of this common organism, and gives insights into the pathogenesis. The unusual dissociation between the clinical presentation and the location of the radiological lesions should also highlight the potential for the condition to mimic the presentation of others, and stimulates debate on the definitions of acute transverse myelitis and acute disseminated encephalomyelitis, and their potential overlap.

  5. Immunization with Pneumococcal Polysaccharide Serotype 3 and Lipopolysaccharide Modulates Lung and Liver Inflammation during a Virulent Streptococcus pneumoniae Infection in Mice

    Restori, Katherine H.; Kennett, Mary J.; Ross, A. Catharine

    2013-01-01

    Vaccination reduces morbidity and mortality from pneumonia, but its effect on the tissue-level response to infection is still poorly understood. We evaluated pneumonia disease progression, acute-phase response, and lung gene expression profiles in mice inoculated intranasally with virulent Gram-positive Streptococcus pneumoniae serotype 3 (ST 3) with and without prior immunization with pneumococcal polysaccharide ST 3 (PPS3) or after coimmunization with PPS3 and a low dose of lipopolysacchari...

  6. Drug Resistance Characteristics and Macrolide-Resistant Mechanisms of Streptococcus pneumoniae in Wenzhou City, China.

    Hu, Dakang; Sun, Zheng; Luo, Xinhua; Liu, Shuangchun; Yu, Lianhua; Qu, Ying; Yang, Jinhong; Yu, Jian; Li, Xiangyang; Zhang, Jin

    2016-01-01

    BACKGROUND Streptococcus pneumoniae (SP) is a Gram-positive, alpha-hemolytic, facultative anaerobic member of the genus Streptococcus. The erythromycin-resistant methylase (erm) gene and macrolide efflux (mef) gene are the 2 main genes that can mediate SP. Transposon (Tn) also plays an important role in the collection and metastasis of the gene. In the present study we investigated the drug resistance characteristics and the macrolide-resistant mechanisms of SP in Wenzhou City, China. MATERIAL AND METHODS Sixty-eight strains of SP were isolated from sputum samples of hospitalized children in the Second Affiliated Hospital of Wenzhou Medical University. These strains were analyzed using antimicrobial susceptibility tests to determine their drug resistance to 10 kinds of antibacterials. Macrolide-resistant phenotypes were identified using K-B method. PCR method was used to analyze the erm B gene, mef A gene, and int Tn gene. RESULTS Drug resistance rates of 68 strains of SP were 98.5%, 100.0%, 63.2%, 52.9%, 94.1%, 89.7%, 0.0%, 0.0%, 16.2%, and 14.7% for clindamycin, erythromycin, penicillin G, cefotaxime, tetracycline, sulfamethoxazole/trimethoprim, levofloxacin, vancomycin, chloramphenicol, and amoxicillin, respectively. Total detection rates of the erm B gene, mef A gene, and int Tn gene were 98.5%, 91.2%, and 100.0%, respectively. CONCLUSIONS SP shows significant multi-drug resistance in Wenzhou City, whereas there is no clinical value of macrolides antibiotics for SP. cMLSB mediated by erm B gene is the most predominant phenotype among macrolide-resistant SP. The int Tn gene may play an important role in horizontal transfer and clonal dissemination of SP drug resistance genes in Wenzhou City. PMID:27483416

  7. Validation of an immunodiagnostic assay for detection of 13 Streptococcus pneumoniae serotype-specific polysaccharides in human urine.

    Pride, Michael W; Huijts, Susanne M; Wu, Kangjian; Souza, Victor; Passador, Sherry; Tinder, Chunyan; Song, Esther; Elfassy, Arik; McNeil, Lisa; Menton, Ronald; French, Roger; Callahan, Janice; Webber, Chris; Gruber, William C; Bonten, Marc J M; Jansen, Kathrin U

    2012-08-01

    To improve the clinical diagnosis of pneumococcal infection in bacteremic and nonbacteremic community-acquired pneumonia (CAP), a Luminex technology-based multiplex urinary antigen detection (UAD) diagnostic assay was developed and validated. The UAD assay can simultaneously detect 13 different serotypes of Streptococcus pneumoniae by capturing serotype-specific S. pneumoniae polysaccharides (PnPSs) secreted in human urine. Assay specificity is achieved by capturing the polysaccharides with serotype-specific monoclonal antibodies (MAbs) on spectrally unique microspheres. Positivity for each serotype was based on positivity cutoff values calculated from a standard curve run on each assay plate together with positive- and negative-control urine samples. The assay is highly specific, since significant signals are detected only when each PnPS was paired with its homologous MAb-coated microspheres. Validation experiments demonstrated excellent accuracy and precision. The UAD assay and corresponding positivity cutoff values were clinically validated by assessing 776 urine specimens obtained from patients with X-ray-confirmed CAP. The UAD assay demonstrated 97% sensitivity and 100% specificity using samples obtained from patients with bacteremic, blood culture-positive CAP. Importantly, the UAD assay identified Streptococcus pneumoniae (13 serotypes) in a proportion of individuals with nonbacteremic CAP, a patient population for which the pneumococcal etiology of CAP was previously difficult to assess. Therefore, the UAD assay provides a specific, noninvasive, sensitive, and reproducible tool to support vaccine efficacy as well as epidemiological evaluation of pneumococcal disease, including CAP, in adults. PMID:22675155

  8. Molecular characteristics of erythromycin-resistant Streptococcus pneumoniae from pediatric patients younger than five years in Beijing, 2010

    Zhou Lin

    2012-10-01

    Full Text Available Abstract Background Streptococcus pneumoniae is the main pathogen that causes respiratory infections in children younger than five years. The increasing incidence of macrolide- and tetracycline-resistant pneumococci among children has been a serious problem in China for many years. The molecular characteristics of erythromycin-resistant pneumococcal isolates that were collected from pediatric patients younger than five years in Beijing in 2010 were analyzed in this study. Results A total of 140 pneumococcal isolates were collected. The resistance rates of all isolates to erythromycin and tetracycline were 96.4% and 79.3%, respectively. Of the 135 erythromycin-resistant pneumococci, 91.1% were non-susceptible to tetracycline. In addition, 30.4% of the erythromycin-resistant isolates expressed both the ermB and mef genes, whereas 69.6% expressed the ermB gene but not the mef gene. Up to 98.5% of the resistant isolates exhibited the cMLSB phenotype, and Tn6002 was the most common transposon present in approximately 56.3% of the resistant isolates, followed by Tn2010, with a proportion of 28.9%. The dominant sequence types (STs in all erythromycin-resistant S. pneumoniae were ST271 (11.9%, ST81 (8.9%, ST876 (8.9%, and ST320 (6.7%, whereas the prevailing serotypes were 19F (19.3%, 23F (9.6%, 14 (9.6%, 15 (8.9%, and 6A (7.4%. The 7-valent pneumococcal conjugate vaccine (PCV7 and 13-valent pneumococcal conjugate vaccine (PCV13 coverage of the erythromycin-resistant pneumococci among the children younger than five years were 45.2% and 62.2%, respectively. ST320 and serotype 19A pneumococci were common in children aged 0 to 2 years. CC271 was the most frequent clonal complex (CC, which accounts for 24.4% of all erythromycin-resistant isolates. Conclusions The non-invasive S. pneumoniae in children younger than five years in Beijing presented high and significant resistance rates to erythromycin and tetracycline. The expressions of ermB and tetM genes

  9. Natural genetic transformation generates a population of merodiploids in Streptococcus pneumoniae.

    Calum Johnston

    Full Text Available Partial duplication of genetic material is prevalent in eukaryotes and provides potential for evolution of new traits. Prokaryotes, which are generally haploid in nature, can evolve new genes by partial chromosome duplication, known as merodiploidy. Little is known about merodiploid formation during genetic exchange processes, although merodiploids have been serendipitously observed in early studies of bacterial transformation. Natural bacterial transformation involves internalization of exogenous donor DNA and its subsequent integration into the recipient genome by homology. It contributes to the remarkable plasticity of the human pathogen Streptococcus pneumoniae through intra and interspecies genetic exchange. We report that lethal cassette transformation produced merodiploids possessing both intact and cassette-inactivated copies of the essential target gene, bordered by repeats (R corresponding to incomplete copies of IS861. We show that merodiploidy is transiently stimulated by transformation, and only requires uptake of a ~3-kb DNA fragment partly repeated in the chromosome. We propose and validate a model for merodiploid formation, providing evidence that tandem-duplication (TD formation involves unequal crossing-over resulting from alternative pairing and interchromatid integration of R. This unequal crossing-over produces a chromosome dimer, resolution of which generates a chromosome with the TD and an abortive chromosome lacking the duplicated region. We document occurrence of TDs ranging from ~100 to ~900 kb in size at various chromosomal locations, including by self-transformation (transformation with recipient chromosomal DNA. We show that self-transformation produces a population containing many different merodiploid cells. Merodiploidy provides opportunities for evolution of new genetic traits via alteration of duplicated genes, unrestricted by functional selective pressure. Transient stimulation of a varied population of

  10. Antibiotic innovation may contribute to slowing the dissemination of multiresistant Streptococcus pneumoniae: the example of ketolides.

    Lulla Opatowski

    Full Text Available BACKGROUND: Despite increasingly frequent bacterial resistance to antibiotics, antibacterial innovation is rare. Ketolides constitute one of the very few new antibiotic classes active against Streptococcus pneumoniae developed during the last 25 years. Their mechanism of action resembles that of macrolides, but they are unaffected by common resistance mechanisms. However, cross-resistance to ketolides has been observed in some macrolide-resistant strains. We examined how new antibiotic exposure may affect overall pneumococcal resistance patterns in the population. The aims of this study were to assess the potential dissemination of newly emerged resistances and to control the selection of strains already multiresistant to existing antimicrobials. METHODOLOGY/PRINCIPAL FINDINGS: We developed an age-structured population model for S. pneumoniae transmission in a human community exposed to heptavalent vaccine, and beta-lactams, macrolides and ketolides. The dynamics of intra-individual selection of resistant strains under antibiotic exposure and interindividual transmission were simulated, with antibiotic-specific resistance mechanisms defining the path to co-resistances and cross-resistances, and parameters concerning the French situation. Results of this simulation study suggest that new antibiotic consumption could markedly slow the diffusion of multiresistant strains. Wider use was associated with slower progression of multiresistance. When ketolides were prescribed to all ages, resistance to them reached 10% after >15 years, while it took >40 years when they were prescribed only to adults. In the scenario according to which new antibiotics totally replaced former antimicrobials, the beta-lactam resistance rate was limited at 70%. CONCLUSIONS: In a context of widespread vaccination and rational use of antibiotics, innovative antibiotic, prescribed to all age groups, may have an added impact on multiresistant-strain dissemination in the

  11. A complex of equine lysozyme and oleic acid with bactericidal activity against Streptococcus pneumoniae.

    Emily A Clementi

    Full Text Available HAMLET and ELOA are complexes consisting of oleic acid and two homologous, yet functionally different, proteins with cytotoxic activities against mammalian cells, with HAMLET showing higher tumor cells specificity, possibly due to the difference in propensity for oleic acid binding, as HAMLET binds 5-8 oleic acid molecules per protein molecule and ELOA binds 11-48 oleic acids. HAMLET has been shown to possess bactericidal activity against a number of bacterial species, particularly those with a respiratory tropism, with Streptococcus pneumoniae displaying the greatest degree of sensitivity. We show here that ELOA also displays bactericidal activity against pneumococci, which at lower concentrations shows mechanistic similarities to HAMLET's bactericidal activity. ELOA binds to S. pneumoniae and causes perturbations of the plasma membrane, including depolarization and subsequent rupture, and activates an influx of calcium into the cells. Selective inhibition of calcium channels and sodium/calcium exchange activity significantly diminished ELOA's bactericidal activity, similar to what we have observed with HAMLET. Finally, ELOA-induced death was also accompanied by DNA fragmentation into high molecular weight fragments - an apoptosis-like morphological phenotype that is seen during HAMLET-induced death. Thus, in contrast to different mechanisms of eukaryote cell death induced by ELOA and HAMLET, these complexes are characterized by rather similar activities towards bacteria. Although the majority of these events could be mimicked using oleic acid alone, the concentrations of oleic acid required were significantly higher than those present in the ELOA complex, and for some assays, the results were not identical between oleic acid alone and the ELOA complex. This indicates that the lipid, as a common denominator in both complexes, is an important component for the complexes' bactericidal activities, while the proteins are required both to solubilize

  12. Reactive Oxygen Species Contribute to the Bactericidal Effects of the Fluoroquinolone Moxifloxacin in Streptococcus pneumoniae.

    Ferrándiz, M J; Martín-Galiano, A J; Arnanz, C; Zimmerman, T; de la Campa, A G

    2016-01-01

    We studied the transcriptomic response of Streptococcus pneumoniae to the fluoroquinolone moxifloxacin at a concentration that inhibits DNA gyrase. Treatment of the wild-type strain R6, at a concentration of 10× the MIC, triggered a response involving 132 genes after 30 min of treatment. Genes from several metabolic pathways involved in the production of pyruvate were upregulated. These included 3 glycolytic enzymes, which ultimately convert fructose 6-phosphate to pyruvate, and 2 enzymes that funnel phosphate sugars into the glycolytic pathway. In addition, acetyl coenzyme A (acetyl-CoA) carboxylase was downregulated, likely leading to an increase in acetyl-CoA. When coupled with an upregulation in formate acetyltransferase, an increase in acetyl-CoA would raise the production of pyruvate. Since pyruvate is converted by pyruvate oxidase (SpxB) into hydrogen peroxide (H2O2), an increase in pyruvate would augment intracellular H2O2. Here, we confirm a 21-fold increase in the production of H2O2 and a 55-fold increase in the amount of hydroxyl radical in cultures treated during 4 h with moxifloxacin. This increase in hydroxyl radical through the Fenton reaction would damage DNA, lipids, and proteins. These reactive oxygen species contributed to the lethality of the drug, a conclusion supported by the observed protective effects of an SpxB deletion. These results support the model whereby fluoroquinolones cause redox alterations. The transcriptional response of S. pneumoniae to moxifloxacin is compared with the response to levofloxacin, an inhibitor of topoisomerase IV. Levofloxacin triggers the transcriptional activation of iron transport genes and also enhances the Fenton reaction. PMID:26525786

  13. The impact of pneumolysin on the macrophage response to Streptococcus pneumoniae is strain-dependent.

    Richard M Harvey

    Full Text Available Streptococcus pneumoniae is the world's leading cause of pneumonia, bacteremia, meningitis and otitis media. A major pneumococcal virulence factor is the cholesterol-dependent cytolysin, which has the defining property of forming pores in cholesterol-containing membranes. In recent times a clinically significant and internationally successful serotype 1 ST306 clone has been found to express a non-cytolytic variant of Ply (Ply306. However, while the pneumococcus is a naturally transformable organism, strains of the ST306 clonal group have to date been virtually impossible to transform, severely restricting efforts to understand the role of non-cytolytic Ply in the success of this clone. In this study isogenic Ply mutants were constructed in the D39 background and for the first time in the ST306 background (A0229467 to enable direct comparisons between Ply variants for their impact on the immune response in a macrophage-like cell line. Strains that expressed cytolytic Ply were found to induce a significant increase in IL-1β release from macrophage-like cells compared to the non-cytolytic and Ply-deficient strains in a background-independent manner, confirming the requirement for pore formation in the Ply-dependent activation of the NLRP3 inflammasome. However, cytolytic activity in the D39 background was found to induce increased expression of the genes encoding GM-CSF (CSF2, p19 subunit of IL-23 (IL23A and IFNβ (IFNB1 compared to non-cytolytic and Ply-deficient D39 mutants, but had no effect in the A0229467 background. The impact of Ply on the immune response to the pneumococcus is highly dependent on the strain background, thus emphasising the importance of the interaction between specific virulence factors and other components of the genetic background of this organism.

  14. Epigenetic Switch Driven by DNA Inversions Dictates Phase Variation in Streptococcus pneumoniae

    Wang, Juanjuan; An, Haoran; Liu, Yanni; Wang, Kailing; Miao, Zhun; Liang, Wenbo; Sebra, Robert; Wang, Guilin; Wang, Wen-Ching; Zhang, Jing-Ren

    2016-01-01

    DNA methylation is an important epigenetic mechanism for phenotypic diversification in all forms of life. We previously described remarkable cell-to-cell heterogeneity in epigenetic pattern within a clonal population of Streptococcus pneumoniae, a leading human pathogen. We here report that the epigenetic diversity is caused by extensive DNA inversions among hsdSA, hsdSB, and hsdSC, three methyltransferase hsdS genes in the Spn556II type-I restriction modification (R-M) locus. Because hsdSA encodes the sequence recognition subunit of this type-I R-M DNA methyltransferase, these site-specific recombinations generate pneumococcal cells with variable HsdSA alleles and thereby diverse genome methylation patterns. Most importantly, the DNA methylation pattern specified by the HsdSA1 allele leads to the formation of opaque colonies, whereas the pneumococci lacking HsdSA1 produce transparent colonies. Furthermore, this HsdSA1-dependent phase variation requires intact DNA methylase activity encoded by hsdM in the Spn556II (renamed colony opacity determinant or cod) locus. Thus, the DNA inversion-driven ON/OFF switch of the hsdSA1 allele in the cod locus and resulting epigenetic switch dictate the phase variation between the opaque and transparent phenotypes. Phase variation has been well documented for its importance in pneumococcal carriage and invasive infection, but its molecular basis remains unclear. Our work has discovered a novel epigenetic cause for this significant pathobiology phenomenon in S. pneumoniae. Lastly, our findings broadly represents a significant advancement in our understanding of bacterial R-M systems and their potential in shaping epigenetic and phenotypic diversity of the prokaryotic organisms because similar site-specific recombination systems widely exist in many archaeal and bacterial species. PMID:27427949

  15. Decrease in penicillin susceptibility due to heat shock protein ClpL in Streptococcus pneumoniae.

    Tran, Thao Dang-Hien; Kwon, Hyog-Young; Kim, Eun-Hye; Kim, Ki-Woo; Briles, David E; Pyo, Suhkneung; Rhee, Dong-Kwon

    2011-06-01

    Antibiotic resistance and tolerance are increasing threats to global health as antibiotic-resistant bacteria can cause severe morbidity and mortality and can increase treatment cost 10-fold. Although several genes contributing to antibiotic tolerance among pneumococci have been identified, we report here that ClpL, a major heat shock protein, could modulate cell wall biosynthetic enzymes and lead to decreased penicillin susceptibility. On capsular type 1, 2, and 19 genetic backgrounds, mutants lacking ClpL were more susceptible to penicillin and had thinner cell walls than the parental strains, whereas a ClpL-overexpressing strain showed a higher resistance to penicillin and a thicker cell wall. Although exposure of Streptococcus pneumoniae D39 to penicillin inhibited expression of the major cell wall synthesis gene pbp2x, heat shock induced a ClpL-dependent increase in the mRNA levels and protein synthesized by pbp2x. Inducible ClpL expression correlated with PBP2x expression and penicillin susceptibility. Fractionation and electron micrograph data revealed that ClpL induced by heat shock is localized at the cell wall, and the ΔclpL showed significantly reduced net translocation of PBP2x into the cell wall. Moreover, coimmunoprecipitation with either ClpL or PBP2x antibody followed by reprobing with ClpL or PBP2x antibody showed an interaction between ClpL and PBP2x after heat stress. This interaction was confirmed by His tag pulldown assay with either ClpLHis₆ or PBP2xHis₆. Thus, ClpL stabilized pbp2x expression, interacted with PBP2x, and facilitated translocation of PBP2x, a key protein of cell wall synthesis process, contributing to the decrease of antibiotic susceptibility in S. pneumoniae. PMID:21422206

  16. Clinical presentation and prognostic factors of Streptococcus pneumoniae meningitis according to the focus of infection

    Samuelsson Susanne

    2005-10-01

    Full Text Available Abstract Background We conducted a nationwide study in Denmark to identify clinical features and prognostic factors in patients with Streptococcus pneumoniae according to the focus of infection. Methods Based on a nationwide registration, clinical information's was prospectively collected from all reported cases of pneumococcal meningitis during a 2-year period (1999–2000. Clinical and laboratory findings at admission, clinical course and outcome of the disease including follow-up audiological examinations were collected retrospectively. The focus of infection was determined according to the clinical diagnosis made by the physicians and after review of the medical records. Results 187 consecutive cases with S. pneumoniae meningitis were included in the study. The most common focus was ear (30%, followed by lung (18%, sinus (8%, and other (2%. In 42% of cases a primary infection focus could not be determined. On admission, fever and an altered mental status were the most frequent findings (in 93% and 94% of cases, respectively, whereas back rigidity, headache and convulsion were found in 57%, 41% and 11% of cases, respectively. 21% of patients died during hospitalisation (adults: 27% vs. children: 2%, Fisher Exact Test, P P = 0.0005. Prognostic factors associated with fatal outcome in univariate logistic regression analysis were advanced age, presence of an underlying disease, history of headache, presence of a lung focus, absence of an otogenic focus, having a CT-scan prior to lumbar puncture, convulsions, requirement of assisted ventilation, and alterations in various CSF parameters (WBC P P = 0.005. Conclusion These results emphasize the prognostic importance of an early recognition of a predisposing focus to pneumococcal meningitis.

  17. Epigenetic Switch Driven by DNA Inversions Dictates Phase Variation in Streptococcus pneumoniae.

    Li, Jing; Li, Jing-Wen; Feng, Zhixing; Wang, Juanjuan; An, Haoran; Liu, Yanni; Wang, Yang; Wang, Kailing; Zhang, Xuegong; Miao, Zhun; Liang, Wenbo; Sebra, Robert; Wang, Guilin; Wang, Wen-Ching; Zhang, Jing-Ren

    2016-07-01

    DNA methylation is an important epigenetic mechanism for phenotypic diversification in all forms of life. We previously described remarkable cell-to-cell heterogeneity in epigenetic pattern within a clonal population of Streptococcus pneumoniae, a leading human pathogen. We here report that the epigenetic diversity is caused by extensive DNA inversions among hsdSA, hsdSB, and hsdSC, three methyltransferase hsdS genes in the Spn556II type-I restriction modification (R-M) locus. Because hsdSA encodes the sequence recognition subunit of this type-I R-M DNA methyltransferase, these site-specific recombinations generate pneumococcal cells with variable HsdSA alleles and thereby diverse genome methylation patterns. Most importantly, the DNA methylation pattern specified by the HsdSA1 allele leads to the formation of opaque colonies, whereas the pneumococci lacking HsdSA1 produce transparent colonies. Furthermore, this HsdSA1-dependent phase variation requires intact DNA methylase activity encoded by hsdM in the Spn556II (renamed colony opacity determinant or cod) locus. Thus, the DNA inversion-driven ON/OFF switch of the hsdSA1 allele in the cod locus and resulting epigenetic switch dictate the phase variation between the opaque and transparent phenotypes. Phase variation has been well documented for its importance in pneumococcal carriage and invasive infection, but its molecular basis remains unclear. Our work has discovered a novel epigenetic cause for this significant pathobiology phenomenon in S. pneumoniae. Lastly, our findings broadly represents a significant advancement in our understanding of bacterial R-M systems and their potential in shaping epigenetic and phenotypic diversity of the prokaryotic organisms because similar site-specific recombination systems widely exist in many archaeal and bacterial species. PMID:27427949

  18. Serotype distribution of Streptococcus pneumoniae in children with invasive diseases in Turkey: 2008-2014.

    Ceyhan, Mehmet; Ozsurekci, Yasemin; Gürler, Nezahat; Öksüz, Lütfiye; Aydemir, Sohret; Ozkan, Sengul; Yuksekkaya, Serife; Keser Emiroglu, Melike; Gültekin, Meral; Yaman, Akgün; Kiremitci, Abdurrahman; Yanık, Keramettin; Karli, Arzu; Ozcinar, Hatice; Aydin, Faruk; Bayramoglu, Gulcin; Zer, Yasemin; Gulay, Zeynep; Gayyurhan, Efgan Dogan; Gül, Mustafa; Özakın, Cüneyt; Güdücüoğlu, Hüseyin; Perçin, Duygu; Akpolat, Nezahat; Ozturk, Candan; Camcıoğlu, Yıldız; Karadağ Öncel, Eda; Çelik, Melda; Şanal, Laser; Uslu, Hakan

    2016-01-01

    Successful vaccination policies for protection from invasive pneumococcal diseases (IPD) dependent on determination of the exact serotype distribution in each country. We aimed to identify serotypes of pneumococcal strains causing IPD in children in Turkey and emphasize the change in the serotypes before and after vaccination with 7-valent pneumococcal conjugate vaccine (PCV-7) was included and PCV-13 was newly changed in Turkish National Immunization Program. Streptococcus pneumoniae strains were isolated at 22 different hospitals of Turkey, which provide healthcare services to approximately 65% of the Turkish population. Of the 335 diagnosed cases with S. pneumoniae over the whole period of 2008-2014, the most common vaccine serotypes were 19F (15.8%), 6B (5.9%), 14 (5.9%), and 3 (5.9%). During the first 5 y of age, which is the target population for vaccination, the potential serotype coverage ranged from 57.5 % to 36.8%, from 65.0% to 44.7%, and from 77.4% to 60.5% for PCV-7, PCV-10, and PCV-13 in 2008-2014, respectively. The ratio of non-vaccine serotypes was 27.2% in 2008-2010 whereas was 37.6% in 2011-2014 (p=0.045). S. penumoniae serotypes was less non-susceptible to penicillin as compared to our previous results (33.7 vs 16.5 %, p=0.001). The reduction of those serotype coverage in years may be attributed to increasing vaccinated children in Turkey and the increasing non-vaccine serotype may be explained by serotype replacement. Our ongoing IPD surveillance is a significant source of information for the decision-making processes on pneumococcal vaccination. PMID:26325175

  19. NanA, a Neuraminidase from Streptococcus pneumoniae, Shows High Levels of Sequence Diversity, at Least in Part through Recombination with Streptococcus oralis

    King, Samantha J.; Whatmore, Adrian M.; Dowson, Christopher G.

    2005-01-01

    Streptococcus pneumoniae, an important human pathogen, contains at least two genes, nanA and nanB, that express sialidase activity. NanA is a virulence determinant of pneumococci which is important in animal models of colonization and middle ear infections. The gene encoding NanA was detected in all 106 pneumococcal strains screened that represented 59 restriction profiles. Sequencing confirmed a high level of diversity, up to 17.2% at the nucleotide level and 14.8% at the amino acid level. N...

  20. One-step multiplex PCR assay for detecting Streptococcus pneumoniae serogroups/types covered by 13-valent pneumococcal conjugate vaccine (PCV13.

    Fatma Filiz Coskun-Ari

    Full Text Available The life-threatening illnesses caused by Streptococcus pneumoniae have been declined significantly after the use of pneumococcal conjugate vaccines. Continuous monitoring of the vaccine serogroups/types is necessary to follow the changing epidemiology of invasive pneumococcal diseases. Recently, the sequential multiplex PCR approach, which uses several different sets of reactions, has been commonly adopted for determining capsular serogroups/types of S. pneumoniae isolates. In our study, we focused on development of a one-step multiplex PCR assay detecting all 1, 3, 4, 5, 6A/B, 7F, 9V, 14, 18C, 19A, 19F and 23F serogroups/types targeted by PCV13. The content of multiplex PCR mix and the cycling conditions were optimized in a manner that allowed rapid and accurate serotyping of a pneumococcal isolate by performing only a single amplification reaction. In our study of 182 clinical isolates, the one-step multiplex PCR assay exhibited 100% sensitivity and specificity, suggesting that its utilization can significantly reduce the use of traditional antiserum method requiring expensive reagents.

  1. Control of competence by related non-coding csRNAs in Streptococcus pneumoniae R6.

    Anke eLaux

    2015-07-01

    Full Text Available The two-component regulatory system CiaRH of Streptococcus pneumoniae is involved in ß-lactam resistance, maintenance of cell integrity, bacteriocin production, host colonization, virulence, and competence. The response regulator CiaR controls, among other genes, expression of five highly similar small non-coding RNAs, designated csRNAs. These csRNAs control competence development by targeting comC, encoding the precursor of the competence stimulating peptide CSP, which is essential to initiate the regulatory cascade leading to competence. In addition, another gene product of the CiaR regulon, the serine protease HtrA, is also involved in competence control. In the absence of HtrA, five csRNAs could suppress competence, but one csRNA alone was not effective. To determine if all csRNAs are needed, reporter gene fusions to competence genes were used to monitor competence gene expression in the presence of different csRNAs. These experiments showed that two csRNAs were not enough to prevent competence, but combinations of three csRNAs, csRNA1,2, 3, or csRNA1,2,4 were sufficient. In S. pneumoniae strains expressing only csRNA5, a surprising positive effect was detected on the level of early competence gene expression. Hence, the role of the csRNAs in competence regulation is more complex than anticipated. Mutations in comC (comC8 partially disrupting predicted complementarity to the csRNAs led to competence even in the presence of all csRNAs. Reconstitution of csRNA complementarity to comC8 restored competence suppression. Again, more than one csRNA was needed. In this case, even two mutated csRNAs complementary to comC8, csRNA1-8 and csRNA2-8, were suppressive. In conclusion, competence in S. pneumoniae is additively controlled by the csRNAs via post-transcriptional regulation of comC.

  2. Post-operative Streptococcus pneumoniae meningoencephalitis complicating surgery for acromegaly in an identical twin.

    Cote, David J; Iuliano, Sherry L; Smith, Timothy R; Laws, Edward R

    2015-06-01

    This case report provides provocative and useful data regarding two aspects of acromegaly and its management. The patient, who is one of a pair of identical twins, has no known hereditary, genetic or otherwise potentially etiologic factors as compared to her unaffected sister. Secondly, transsphenoidal surgery, which was ultimately successful, was complicated by pneumococcal meningitis, an unusual event with only four previously reported patients, three of whom ended in death or major neurologic deficits. In this case, a 57-year-old woman gradually developed classical signs and symptoms of acromegaly while her identical twin sister remained normal with no evidence of endocrine disease. Endoscopic transsphenoidal surgery was complicated by the development of meningitis 25 days after surgery. This was controlled following a difficult hospital course. Streptococcus pneumoniae meningoencephalitis is a rare but life-threatening complication of transsphenoidal surgery. A high index of suspicion for incipient meningitis should be maintained when patients present with severe headache and increased intracranial pressure, even if they initially lack the typical symptoms and signs. Immediate and aggressive treatment is necessary to avoid significant neurologic deficit. PMID:25861890

  3. Modeling transmission of multitype infectious agents: application to carriage of Streptococcus pneumoniae.

    Erästö, Panu; Hoti, Fabian; Auranen, Kari

    2012-06-30

    We describe a novel Bayesian approach to estimate acquisition and clearance rates for many competing subtypes of a pathogen in a susceptible-infected-susceptible model. The inference relies on repeated measurements of the current status of being a non-carrier (susceptible) or a carrier (infected) of one of the n(q)  > 1 subtypes. We typically collect the measurements with sampling intervals that may not catch the true speed of the underlying dynamics. We tackle the problem of incompletely observed data with Bayesian data augmentation, which integrates over possible carriage histories, allowing the data to contain intermittently missing values, complete dropouts of study subjects, or inclusion of new study subjects during the follow-up. We investigate the performance of the described method through simulations by using two different mixing groups (family and daycare) and different sampling intervals. For comparison, we describe crude maximum likelihood-based estimates derived directly from the observations. We apply the estimation algorithm to data about transmission of Streptococcus pneumonia in Bangladeshi families. The computationally intensive Bayesian approach is a valid method to account for incomplete observations, and we found that it performs generally better than the simple crude method, in particular with large amount of missing data. PMID:22354452

  4. Coordinated Bacteriocin Expression and Competence in Streptococcus pneumoniae Contributes to Genetic Adaptation through Neighbor Predation.

    Wei-Yun Wholey

    2016-02-01

    Full Text Available Streptococcus pneumoniae (pneumococcus has remained a persistent cause of invasive and mucosal disease in humans despite the widespread use of antibiotics and vaccines. The resilience of this organism is due to its capacity for adaptation through the uptake and incorporation of new genetic material from the surrounding microbial community. DNA uptake and recombination is controlled by a tightly regulated quorum sensing system that is triggered by the extracellular accumulation of competence stimulating peptide (CSP. In this study, we demonstrate that CSP can stimulate the production of a diverse array of blp bacteriocins. This cross stimulation occurs through increased production and secretion of the bacteriocin pheromone, BlpC, and requires a functional competence regulatory system. We show that a highly conserved motif in the promoter of the operon encoding BlpC and its transporter mediates the upregulation by CSP. The accumulation of BlpC following CSP stimulation results in augmented activation of the entire blp locus. Using biofilm-grown organisms as a model for competition and genetic exchange on the mucosal surface, we demonstrate that DNA exchange is enhanced by bacteriocin secretion suggesting that co-stimulation of bacteriocins with competence provides an adaptive advantage. The blp and com regulatory pathways are believed to have diverged and specialized in a remote ancestor of pneumococcus. Despite this, the two systems have maintained a regulatory connection that promotes competition and adaptation by targeting for lysis a wide array of potential competitors while simultaneously providing the means for incorporation of their DNA.

  5. A Multi-Scale Computational Study on the Mechanism of Streptococcus pneumoniae Nicotinamidase (SpNic

    Bogdan F. Ion

    2014-09-01

    Full Text Available Nicotinamidase (Nic is a key zinc-dependent enzyme in NAD metabolism that catalyzes the hydrolysis of nicotinamide to give nicotinic acid. A multi-scale computational approach has been used to investigate the catalytic mechanism, substrate binding and roles of active site residues of Nic from Streptococcus pneumoniae (SpNic. In particular, density functional theory (DFT, molecular dynamics (MD and ONIOM quantum mechanics/molecular mechanics (QM/MM methods have been employed. The overall mechanism occurs in two stages: (i formation of a thioester enzyme-intermediate (IC2 and (ii hydrolysis of the thioester bond to give the products. The polar protein environment has a significant effect in stabilizing reaction intermediates and in particular transition states. As a result, both stages effectively occur in one step with Stage 1, formation of IC2, being rate limiting barrier with a cost of 53.5 kJ•mol−1 with respect to the reactant complex, RC. The effects of dispersion interactions on the overall mechanism were also considered but were generally calculated to have less significant effects with the overall mechanism being unchanged. In addition, the active site lysyl (Lys103 is concluded to likely play a role in stabilizing the thiolate of Cys136 during the reaction.

  6. A functional genomics approach to establish the complement of carbohydrate transporters in Streptococcus pneumoniae.

    Alessandro Bidossi

    Full Text Available The aerotolerant anaerobe Streptococcus pneumoniae is part of the normal nasopharyngeal microbiota of humans and one of the most important invasive pathogens. A genomic survey allowed establishing the occurrence of twenty-one phosphotransferase systems, seven carbohydrate uptake ABC transporters, one sodium:solute symporter and a permease, underlining an exceptionally high capacity for uptake of carbohydrate substrates. Despite high genomic variability, combined phenotypic and genomic analysis of twenty sequenced strains did assign the substrate specificity only to two uptake systems. Systematic analysis of mutants for most carbohydrate transporters enabled us to assign a phenotype and substrate specificity to twenty-three transport systems. For five putative transporters for galactose, pentoses, ribonucleosides and sulphated glycans activity was inferred, but not experimentally confirmed and only one transport system remains with an unknown substrate and lack of any functional annotation. Using a metabolic approach, 80% of the thirty-two fermentable carbon substrates were assigned to the corresponding transporter. The complexity and robustness of sugar uptake is underlined by the finding that many transporters have multiple substrates, and many sugars are transported by more than one system. The present work permits to draw a functional map of the complete arsenal of carbohydrate utilisation proteins of pneumococci, allows re-annotation of genomic data and might serve as a reference for related species. These data provide tools for specific investigation of the roles of the different carbon substrates on pneumococcal physiology in the host during carriage and invasive infection.

  7. A Functional Genomics Approach to Establish the Complement of Carbohydrate Transporters in Streptococcus pneumoniae

    Bidossi, Alessandro; Mulas, Laura; Decorosi, Francesca; Colomba, Leonarda; Ricci, Susanna; Pozzi, Gianni; Deutscher, Josef; Viti, Carlo; Oggioni, Marco Rinaldo

    2012-01-01

    The aerotolerant anaerobe Streptococcus pneumoniae is part of the normal nasopharyngeal microbiota of humans and one of the most important invasive pathogens. A genomic survey allowed establishing the occurrence of twenty-one phosphotransferase systems, seven carbohydrate uptake ABC transporters, one sodium∶solute symporter and a permease, underlining an exceptionally high capacity for uptake of carbohydrate substrates. Despite high genomic variability, combined phenotypic and genomic analysis of twenty sequenced strains did assign the substrate specificity only to two uptake systems. Systematic analysis of mutants for most carbohydrate transporters enabled us to assign a phenotype and substrate specificity to twenty-three transport systems. For five putative transporters for galactose, pentoses, ribonucleosides and sulphated glycans activity was inferred, but not experimentally confirmed and only one transport system remains with an unknown substrate and lack of any functional annotation. Using a metabolic approach, 80% of the thirty-two fermentable carbon substrates were assigned to the corresponding transporter. The complexity and robustness of sugar uptake is underlined by the finding that many transporters have multiple substrates, and many sugars are transported by more than one system. The present work permits to draw a functional map of the complete arsenal of carbohydrate utilisation proteins of pneumococci, allows re-annotation of genomic data and might serve as a reference for related species. These data provide tools for specific investigation of the roles of the different carbon substrates on pneumococcal physiology in the host during carriage and invasive infection. PMID:22428019

  8. Mutational and Biochemical Analysis of the DNA-entry Nuclease EndA from Streptococcus pneumoniae

    M Midon; P Schafer; A Pingoud; M Ghosh; A Moon; M Cuneo; R London; G Meiss

    2011-12-31

    EndA is a membrane-attached surface-exposed DNA-entry nuclease previously known to be required for genetic transformation of Streptococcus pneumoniae. More recent studies have shown that the enzyme also plays an important role during the establishment of invasive infections by degrading extracellular chromatin in the form of neutrophil extracellular traps (NETs), enabling streptococci to overcome the innate immune system in mammals. As a virulence factor, EndA has become an interesting target for future drug design. Here we present the first mutational and biochemical analysis of recombinant forms of EndA produced either in a cell-free expression system or in Escherichia coli. We identify His160 and Asn191 to be essential for catalysis and Asn182 to be required for stability of EndA. The role of His160 as the putative general base in the catalytic mechanism is supported by chemical rescue of the H160A variant of EndA with imidazole added in excess. Our study paves the way for the identification and development of protein or low-molecular-weight inhibitors for EndA in future high-throughput screening assays.

  9. Transport of Streptococcus pneumoniae capsular polysaccharide in MHC Class II tubules.

    Tom Li Stephen

    2007-03-01

    Full Text Available Bacterial capsular polysaccharides are virulence factors and are considered T cell-independent antigens. However, the capsular polysaccharide Sp1 from Streptococcus pneumoniae serotype 1 has been shown to activate CD4(+ T cells in a major histocompatibility complex (MHC class II-dependent manner. The mechanism of carbohydrate presentation to CD4(+ T cells is unknown. We show in live murine dendritic cells (DCs that Sp1 translocates from lysosomal compartments to the plasma membrane in MHCII-positive tubules. Sp1 cell surface presentation results in reduction of self-peptide presentation without alteration of the MHCII self peptide repertoire. In DM-deficient mice, retrograde transport of Sp1/MHCII complexes resulting in T cell-dependent immune responses to the polysaccharide in vitro and in vivo is significantly reduced. The results demonstrate the capacity of a bacterial capsular polysaccharide antigen to use DC tubules as a vehicle for its transport as an MHCII/saccharide complex to the cell surface for the induction of T cell activation. Furthermore, retrograde transport requires the functional role of DM in self peptide-carbohydrate exchange. These observations open new opportunities for the design of vaccines against microbial encapsulated pathogens.

  10. Serotypes and antimicrobial resistance of meningeal isolates of Streptococcus pneumonia. Cuba, 2007-2012

    Gilda Toraño-Peraza

    2014-12-01

    Full Text Available An observational study was conducted to know the serotypes and antimicrobial susceptibility of isolates of Streptococcus pneumoniae responsible for meningitis in Cuba, where there is no vaccine yet to prevent invasive pneumococcal disease. The study included the total number of isolates submitted to the "Pedro Kourí" Institute between 2007 and 2012 (N=237. Serotypes identification was performed using capsular swelling test and antimicrobial susceptibility was studied by determining the minimum inhibitory concentration using the broth microdilution method. Predominant serotypes were 6A, 6B, 14, 19F and 23F and other non-vaccinal 18 serogroups/serotypes were identified in 29.1% of the isolates. A tendency to an increased resistance to penicillin (44.3 % was observed; the most common resistance patterns were: penicillin-trimethoprim/sulfamethoxazole and penicillin-erythromycin (21.1% and 10.5%, respectively. The largest number of isolates resistant to penicillin was in serotypes 6B, 14, 19F and 23F and the possibility of resistant non-vaccine serotypes emergence should be considered. The results show that 70.4 % of the isolates studied corresponds to the serotypes included in 13-valent conjugated pneumococcal vaccine, but with 10-valent it would achieve a lower vaccination potential coverage (56.1%. This information must be considered when evaluating the decision to use in Cuba any commercially available vaccine or the proposal of another strategy of vaccination from autochthonous vaccine candidates.

  11. Mechanism of β-lactam action in Streptococcus pneumoniae: the piperacillin paradox.

    Philippe, Jules; Gallet, Benoit; Morlot, Cécile; Denapaite, Dalia; Hakenbeck, Regine; Chen, Yuxin; Vernet, Thierry; Zapun, André

    2015-01-01

    The human pathogen Streptococcus pneumoniae has been treated for decades with β-lactam antibiotics. Its resistance is now widespread, mediated by the expression of mosaic variants of the target enzymes, the penicillin-binding proteins (PBPs). Understanding the mode of action of β-lactams, not only in molecular detail but also in their physiological consequences, will be crucial to improving these drugs and any counterresistances. In this work, we investigate the piperacillin paradox, by which this β-lactam selects primarily variants of PBP2b, whereas its most reactive target is PBP2x. These PBPs are both essential monofunctional transpeptidases involved in peptidoglycan assembly. PBP2x participates in septal synthesis, while PBP2b functions in peripheral elongation. The formation of the "lemon"-shaped cells induced by piperacillin treatment is consistent with the inhibition of PBP2x. Following the examination of treated and untreated cells by electron microscopy, the localization of the PBPs by epifluorescence microscopy, and the determination of the inhibition time course of the different PBPs, we propose a model of peptidoglycan assembly that accounts for the piperacillin paradox. PMID:25385114

  12. Two-component system response regulators involved in virulence of Streptococcus pneumoniae TIGR4 in infective endocarditis.

    Trihn, My; Ge, Xiuchun; Dobson, Alleson; Kitten, Todd; Munro, Cindy L; Xu, Ping

    2013-01-01

    Streptococci resident in the oral cavity have been linked to infective endocarditis (IE). While other viridans streptococci are commonly studied in relation to IE, less research has been focused on Streptococcus pneumoniae. We established for the first time an animal model of S. pneumoniae IE, and examined the virulence of the TIGR4 strain in this model. We hypothesized that two-component systems (TCS) may mediate S. pneumoniae TIGR4 strain virulence in IE and examined TCS response regulator (RR) mutants of TIGR4 in vivo with the IE model. Thirteen of the 14 RR protein genes were mutagenized, excluding only the essential gene SP_1227. The requirement of the 13 RRs for S. pneumoniae competitiveness in the IE model was assessed in vivo through use of quantitative real-time PCR (qPCR) and competitive index assays. Using real-time PCR, several RR mutants were detected at significantly lower levels in infected heart valves compared with a control strain suggesting the respective RRs are candidate virulence factors for IE. The virulence reduction of the ΔciaR mutant was further confirmed by competitive index assay. Our data suggest that CiaR is a virulence factor of S. pneumoniae strain TIGR4 for IE. PMID:23342132

  13. Frequency of Streptococcus pneumonia and Haemophilus influenza in acute exacerbation of chronic obstructive airway disease and their sensitivity to levofloxacin

    Objective: To determine the frequency of Streptococcus pneumoniae and Haemophilus influenzae in acute exacerbation of chronic obstructive pulmonary disease and their sensitivity to levofloxacin. Methods: The cross-sectional study was conducted at the Department of Medicine, AbbasiShaheed Hospital, Karachi, between July 2009 and January 2010. Patients already diagnosed with chronic obstructive pulmonary disease and admitted with symptoms of acute exacerbation were included in the study and their sputum samples were sent for microbiological evaluation. SPSS 16 was used for statistical analysis. Results: Of the total 105 patients in the study, 90 (85.17%) were males. Overall mean age at presentation was 62+-10.2 years. S. pneumoniae was isolated from sputum culture of 33 (31.4%) patients, while 13 (12.4%) patients showed growth of H. influenzae. Out of the 33 sputum specimens of S. pneumoniae, 32 (97.0%) were sensitive to levofloxacin, while 1 (3.0%) was resistant. All the 13 isolates of H. influenzae were sensitive to levofloxacin. Conclusion: S. pneumoniae and H. influenzae are still the most prevalent organisms isolated in acute exacerbation of chronic obstructive pulmonary disease in our population. Levofloxacin is still considered a highly sensitive antibiotic against these common micro-organisms in our population, but S. pneumoniae has started developing resistance against levofloxacin. Therefore, intermittent surveillance regarding development of resistance pattern of common micro-organisms against commonly prescribed antibiotics is required. (author)

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

    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

  15. Enhanced inflammation in aged mice following infection with Streptococcus pneumoniae is associated with decreased IL-10 and augmented chemokine production.

    Williams, Andrew E; José, Ricardo J; Brown, Jeremy S; Chambers, Rachel C

    2015-03-15

    Streptococcus pneumoniae is the most common cause of severe pneumonia in the elderly. However, the impact of aging on the innate inflammatory response to pneumococci is poorly defined. We compared the innate immune response in old vs. young adult mice following infection with S. pneumoniae. The accumulation of neutrophils recovered from bronchoalveolar lavage fluid and lung homogenates was increased in aged compared with young adult mice, although bacterial outgrowth was similar in both age groups, as were markers of microvascular leak. Aged mice had similar levels of IL-1β, TNF, IFN-γ, IL-17, and granulocyte colony-stimulating factor following S. pneumoniae infection, compared with young mice, but increased levels of the chemokines CXCL9, CXCL12, CCL3, CCL4, CCL5, CCL11, and CCL17. Moreover, levels of IL-10 were significantly lower in aged animals. Neutralization of IL-10 in infected young mice was associated with increased neutrophil recruitment but no decrease in bacterial outgrowth. Furthermore, IL-10 neutralization resulted in increased levels of CCL3, CCL5, and CXCL10. We conclude that aging is associated with enhanced inflammatory responses following S. pneumoniae infection as a result of a compromised immunomodulatory cytokine response. PMID:25595646

  16. Clinical Features and Outcomes of Serotype 19A Invasive Pneumococcal Disease in Calgary, Alberta

    Ricketson, Leah J; Otto G Vanderkooi; Wood, Melissa L; Jenine Leal; Kellner, James D

    2014-01-01

    BACKGROUND: Streptoccocus pneumoniae serotype 19A (ST19A) became an important cause of invasive pneumococcal disease (IPD) after the introduction of the conjugate vaccine.OBJECTIVE: To examine the severity and outcome of ST19A IPD compared with non-ST19A IPD.METHODS: The Calgary Area Streptococcus pneumoniae Epidemiology Research (CASPER) study collects clinical and laboratory data on all IPD cases in Calgary, Alberta. Analysis was performed on data from 2000 to 2010 comparing ST19A and non-S...

  17. Detection of lytA, pspC, and rrgA genes in Streptococcus pneumoniae isolated from healthy children

    Gholamhosseini-Moghaddam, Tahereh; Rad, Mehrnaz; Mousavi, Seyed Fazlollah; Ghazvini, Kiarash

    2015-01-01

    Background and Objectives: Many surface proteins are implicated in nasopharyngeal colonization and pathogenesis of Streptococcus pneumoniae. Some of these factors are candidate antigens for protein based vaccines. New vaccine designs focus on the surface proteins (e. g., pspA and pspC) and also cytolysin, and pneumolysin. In this study, 3 key virulence genes, lytA, pspC, and rrgA, which encoded surface proteins, were detected among S. pneumoniae isolates. Materials and Methods: A total of 260 nasopharyngeal swabs were collected from healthy children under 6 years old attending day care centers in Mashhad, Iran. Isolates of S. pneumoniae were confirmed by optochin susceptibility and colony appearance and also by PCR for cpsA gene. The presence of lytA, pspC, and rrgA genes were also detected by PCR. Results: A total of 59 isolates were confirmed as S. pneumoniae. Among these isolates, 50 (84.74%), 19 (32.20%), and 2 (3.38%) were positive for lytA, rrgA, and pspC genes respectively. The presence of these genes among S.pneumoniae isolates were as follows: 1) rrgA, lytA, pspC (1 isolate), 2) rrgA, lytA(17isolates), 3) pspC (2 isolate), 4) lytA (50 isolates). Conclusion: cpsA gene was specific for detection of S. pneumoniae isolates which were colonized in nasopharynx. The lytA gene was the most frequent gene among the S. pneumoniae isolates, and combination of rrgA, lytA was the most observed pattern. Thus, it is important for future monitoring of vaccine formulation in our country. PMID:26668703

  18. Pretreatment of epithelial cells with live Streptococcus pneumoniae has no detectable effect on influenza A virus replication in vitro.

    Kang Ouyang

    Full Text Available Influenza A virus (IAV and Streptococcus pneumoniae (pneumococcus are two major upper respiratory tract pathogens responsible for exacerbated disease in coinfected individuals. Despite several studies showing increased susceptibility to secondary bacterial infections following IAV infection, information on the direct effect of S. pneumoniae on IAV in vitro is unknown. This is an important area of investigation as S. pneumoniae is a common commensal of the human upper respiratory tract, present as an important coinfecting pathogen with IAV infection. A recent study showed that S. pneumoniae enhances human metapneumovirus infection in polarized bronchial epithelial cells in vitro. The aim of the current study was to determine whether treatment of epithelial cells with S. pneumoniae affects IAV replication using a standard immunofluorescence assay (IFA. For this study we used four IAV permissive epithelial cell lines including two human-derived cell lines, 12 pneumococcal strains including recent human clinical isolates which represent different genetic backgrounds and serotypes, and six IAV strains of varying genetic nature and pathogenic potential including the pandemic 2009 H1N1 virus. Our results suggested that pretreatment of MDCK cells with 7.5×10(6 colony-forming units (CFUs of live S. pneumoniae resulted in gradual cell-death in a time-dependent manner (0.5 to 4 hr. But, pretreatment of cell lines with 7.5×10(5 and lower CFUs of S. pneumoniae had no detectable effect on either the morphology of cells or on the IAV replication. However, unlike in epithelial cell lines, due to influence of secreted host factors the effect of pneumococci on IAV replication may be different during coinfections in vivo in the human upper respiratory tract, and in vitro with primary human polarized bronchial epithelial cells.

  19. Identification and characterization of noncoding small RNAs in Streptococcus pneumoniae serotype 2 strain D39.

    Tsui, Ho-Ching Tiffany; Mukherjee, Dhriti; Ray, Valerie A; Sham, Lok-To; Feig, Andrew L; Winkler, Malcolm E

    2010-01-01

    We report a search for small RNAs (sRNAs) in the low-GC, gram-positive human pathogen Streptococcus pneumoniae. Based on bioinformatic analyses by Livny et al. (J. Livny, A. Brencic, S. Lory, and M. K. Waldor, Nucleic Acids Res. 34:3484-3493, 2006), we tested 40 candidates by Northern blotting and confirmed the expression of nine new and one previously reported (CcnA) sRNAs in strain D39. CcnA is one of five redundant sRNAs reported by Halfmann et al. (A. Halfmann, M. Kovacs, R. Hakenbeck, and R. Bruckner, Mol. Microbiol. 66:110-126, 2007) that are positively controlled by the CiaR response regulator. We characterized 3 of these 14 sRNAs: Spd-sr17 (144 nucleotides [nt]; decreased in stationary phase), Spd-sr37 (80 nt; strongly expressed in all growth phases), and CcnA (93 nt; induced by competence stimulatory peptide). Spd-sr17 and CcnA likely fold into structures containing single-stranded regions between hairpin structures, whereas Spd-sr37 forms a base-paired structure. Primer extension mapping and ectopic expression in deletion/insertion mutants confirmed the independent expression of the three sRNAs. Microarray analyses indicated that insertion/deletion mutants in spd-sr37 and ccnA exerted strong cis-acting effects on the transcription of adjacent genes, indicating that these sRNA regions are also cotranscribed in operons. Deletion or overexpression of the three sRNAs did not cause changes in growth, certain stress responses, global transcription, or virulence. Constitutive ectopic expression of CcnA reversed some phenotypes of D39 Delta ciaR mutants, but attempts to link CcnA to -E to comC as a target were inconclusive in ciaR(+) strains. These results show that S. pneumoniae, which lacks known RNA chaperones, expresses numerous sRNAs, but three of these sRNAs do not strongly affect common phenotypes or transcription patterns. PMID:19854910

  20. LuxS mediates iron-dependent biofilm formation, competence, and fratricide in Streptococcus pneumoniae.

    Trappetti, Claudia; Potter, Adam J; Paton, Adrienne W; Oggioni, Marco R; Paton, James C

    2011-11-01

    During infection, Streptococcus pneumoniae exists mainly in sessile biofilms rather than in planktonic form, except during sepsis. The capacity to form biofilms is believed to be important for nasopharyngeal colonization as well as disease pathogenesis, but relatively little is known about the regulation of this process. Here, we investigated the effect of exogenous iron [Fe(III)] as well as the role of luxS (encoding S-ribosylhomocysteine lyase) on biofilm formation by S. pneumoniae D39. Fe(III) strongly enhanced biofilm formation at concentrations of ≥50 μM, while Fe(III) chelation with deferoxamine was inhibitory. Importantly, Fe(III) also upregulated the expression of luxS in wild-type D39. A luxS-deficient mutant (D39luxS) failed to form a biofilm, even with Fe(III) supplementation, whereas a derivative overexpressing luxS (D39luxS+) exhibited enhanced biofilm formation capacity and could form a biofilm without added Fe(III). D39luxS exhibited reduced expression of the major Fe(III) transporter PiuA, and the cellular [Fe(III)] was significantly lower than that in D39; in contrast, D39luxS+ had a significantly higher cellular [Fe(III)] than the wild type. The release of extracellular DNA, which is an important component of the biofilm matrix, also was directly related to luxS expression. Similarly, genetic competence, as measured by transformation frequency as well as the expression of competence genes comD, comX, comW, cglA, and dltA and the murein hydrolase cbpD, which is associated with fratricide-dependent DNA release, all were directly related to luxS expression levels and were further upregulated by Fe(III). Moreover, mutagenesis of cbpD blocked biofilm formation. We propose that competence, fratricide, and biofilm formation are closely linked in pneumococci, and that luxS is a central regulator of these processes. We also propose that the stimulatory effects of Fe(III) on all of these parameters are due to the upregulation of luxS expression, and that

  1. LuxS Mediates Iron-Dependent Biofilm Formation, Competence, and Fratricide in Streptococcus pneumoniae

    Trappetti, Claudia; Potter, Adam J.; Paton, Adrienne W.; Oggioni, Marco R.; Paton, James C.

    2011-01-01

    During infection, Streptococcus pneumoniae exists mainly in sessile biofilms rather than in planktonic form, except during sepsis. The capacity to form biofilms is believed to be important for nasopharyngeal colonization as well as disease pathogenesis, but relatively little is known about the regulation of this process. Here, we investigated the effect of exogenous iron [Fe(III)] as well as the role of luxS (encoding S-ribosylhomocysteine lyase) on biofilm formation by S. pneumoniae D39. Fe(III) strongly enhanced biofilm formation at concentrations of ≥50 μM, while Fe(III) chelation with deferoxamine was inhibitory. Importantly, Fe(III) also upregulated the expression of luxS in wild-type D39. A luxS-deficient mutant (D39luxS) failed to form a biofilm, even with Fe(III) supplementation, whereas a derivative overexpressing luxS (D39luxS+) exhibited enhanced biofilm formation capacity and could form a biofilm without added Fe(III). D39luxS exhibited reduced expression of the major Fe(III) transporter PiuA, and the cellular [Fe(III)] was significantly lower than that in D39; in contrast, D39luxS+ had a significantly higher cellular [Fe(III)] than the wild type. The release of extracellular DNA, which is an important component of the biofilm matrix, also was directly related to luxS expression. Similarly, genetic competence, as measured by transformation frequency as well as the expression of competence genes comD, comX, comW, cglA, and dltA and the murein hydrolase cbpD, which is associated with fratricide-dependent DNA release, all were directly related to luxS expression levels and were further upregulated by Fe(III). Moreover, mutagenesis of cbpD blocked biofilm formation. We propose that competence, fratricide, and biofilm formation are closely linked in pneumococci, and that luxS is a central regulator of these processes. We also propose that the stimulatory effects of Fe(III) on all of these parameters are due to the upregulation of luxS expression, and that

  2. Competence for genetic transformation in Streptococcus pneumoniae: mutations in σA bypass the comW requirement.

    Tovpeko, Yanina; Morrison, Donald A

    2014-11-01

    Competence for genetic transformation in the genus Streptococcus depends on an alternative sigma factor, σ(X), for coordinated synthesis of 23 proteins, which together establish the X state by permitting lysis of incompetent streptococci, uptake of DNA fragments, and integration of strands of that DNA into the resident genome. Initiation of transient accumulation of high levels of σ(X) is coordinated between cells by transcription factors linked to peptide pheromone signals. In Streptococcus pneumoniae, elevated σ(X) is insufficient for development of full competence without coexpression of a second competence-specific protein, ComW. ComW, shared by eight species in the Streptococcus mitis and Streptococcus anginosus groups, is regulated by the same pheromone circuit that controls σ(X), but its role in expression of the σ(X) regulon is unknown. Using the strong, but not absolute, dependence of transformation on comW as a selective tool, we collected 27 independent comW bypass mutations and mapped them to 10 single-base transitions, all within rpoD, encoding the primary sigma factor subunit of RNA polymerase, σ(A). Eight mapped to sites in rpoD region 4 that are implicated in interaction with the core β subunit, indicating that ComW may act to facilitate competition of the alternative sigma factor σ(X) for access to core polymerase. PMID:25112479

  3. The structural basis for T-antigen hydrolysis by Streptococcus pneumoniae: a target for structure-based vaccine design.

    Caines, Matthew E C; Zhu, Haizhong; Vuckovic, Marija; Willis, Lisa M; Withers, Stephen G; Wakarchuk, Warren W; Strynadka, Natalie C J

    2008-11-14

    Streptococcus pneumoniae endo-alpha-N-acetylgalactosaminidase is a cell surface-anchored glycoside hydrolase from family GH101 involved in the breakdown of mucin type O-linked glycans. The 189-kDa mature enzyme specifically hydrolyzes the T-antigen disaccharide from extracellular host glycoproteins and is representative of a broadly important class of virulence factors that have remained structurally uncharacterized due to their large size and highly modular nature. Here we report a 2.9 angstroms resolution crystal structure that remarkably captures the multidomain architecture and characterizes a catalytic center unexpectedly resembling that of alpha-amylases. Our analysis presents a complete model of glycoprotein recognition and provides a basis for the structure-based design of novel Streptococcus vaccines and therapeutics. PMID:18784084

  4. Role for Toll-like receptor 2 in the immune response to Streptococcus pneumoniae infection in mouse otitis media.

    Han, Fengchan; Yu, Heping; Tian, Cong; Li, Shengli; Jacobs, Michael R; Benedict-Alderfer, Cindy; Zheng, Qing Y

    2009-07-01

    Streptococcus pneumoniae is the most common pathogen associated with otitis media. To examine the role of Toll-like receptor 2 (TLR2) in host defense against Streptococcus pneumoniae infection in the middle ear, wild-type (WT; C57BL/6) and TLR2-deficient (TLR2(-/-)) mice were inoculated with Streptococcus pneumoniae (1 x 10(6) CFU) through the tympanic membrane. Nineteen of 37 TLR2(-/-) mice showed bacteremia and died within 3 days after the challenge, compared to only 4 of 32 WT mice that died. Of those that survived, more severe hearing loss in the TLR2(-/-) mice than in the WT mice was indicated by an elevation in auditory-evoked brain stem response thresholds at 3 or 7 days postinoculation. The histological pathology was characterized by effusion and tissue damage in the middle ear, and in the TLR2(-/-) mice, the outcome of infection became more severe at 7 days. At both 3 and 7 days postchallenge, the TLR2(-/-) mice had higher blood bacterial titers than the WT mice (P interleukin 1beta, MIP1alpha, Muc5ac, and Muc5b were significantly lower in the ears of TLR2(-/-) mice than in WT mice. In summary, TLR2(-/-) mice may produce relatively low levels of proinflammatory cytokines following pneumococcal challenge, thus hindering the clearance of bacteria from the middle ear and leading to sepsis and a high mortality rate. This study provides evidence that TLR2 is important in the molecular pathogenesis and host response to otitis media. PMID:19414550

  5. Distinct effects on diversifying selection by two mechanisms of immunity against Streptococcus pneumoniae.

    Yuan Li

    Full Text Available Antigenic variation to evade host immunity has long been assumed to be a driving force of diversifying selection in pathogens. Colonization by Streptococcus pneumoniae, which is central to the organism's transmission and therefore evolution, is limited by two arms of the immune system: antibody- and T cell- mediated immunity. In particular, the effector activity of CD4(+ T(H17 cell mediated immunity has been shown to act in trans, clearing co-colonizing pneumococci that do not bear the relevant antigen. It is thus unclear whether T(H17 cell immunity allows benefit of antigenic variation and contributes to diversifying selection. Here we show that antigen-specific CD4(+ T(H17 cell immunity almost equally reduces colonization by both an antigen-positive strain and a co-colonized, antigen-negative strain in a mouse model of pneumococcal carriage, thus potentially minimizing the advantage of escape from this type of immunity. Using a proteomic screening approach, we identified a list of candidate human CD4(+ T(H17 cell antigens. Using this list and a previously published list of pneumococcal Antibody antigens, we bioinformatically assessed the signals of diversifying selection among the identified antigens compared to non-antigens. We found that Antibody antigen genes were significantly more likely to be under diversifying selection than the T(H17 cell antigen genes, which were indistinguishable from non-antigens. Within the Antibody antigens, epitopes recognized by human antibodies showed stronger evidence of diversifying selection. Taken together, the data suggest that T(H17 cell-mediated immunity, one form of T cell immunity that is important to limit carriage of antigen-positive pneumococcus, favors little diversifying selection in the targeted antigen. The results could provide new insight into pneumococcal vaccine design.

  6. Co-Transcriptomes of Initial Interactions In Vitro between Streptococcus Pneumoniae and Human Pleural Mesothelial Cells.

    Claire J Heath

    Full Text Available Streptococcus pneumoniae (Spn is a major causative organism of empyema, an inflammatory condition occurring in the pleural sac. In this study, we used human and Spn cDNA microarrays to characterize the transcriptional responses occurring during initial contact between Spn and a human pleural mesothelial cell line (PMC in vitro. Using stringent filtering criteria, 42 and 23 Spn genes were up-and down-regulated respectively. In particular, genes encoding factors potentially involved in metabolic processes and Spn adherence to eukaryotic cells were up-regulated e.g. glnQ, glnA, aliA, psaB, lytB and nox. After Spn initial contact, 870 human genes were differentially regulated and the largest numbers of significant gene expression changes were found in canonical pathways for eukaryotic initiation factor 2 signaling (60 genes out of 171, oxidative phosphorylation (32/103, mitochondrial dysfunction (37/164, eIF4 and p70S6K signaling (28/142, mTOR signaling (27/182, NRF2-mediated oxidative stress response (20/177, epithelial adherens junction remodeling (11/66 and ubiquitination (22/254. The cellular response appeared to be directed towards host cell survival and defense. Spn did not activate NF-kB or phosphorylate p38 MAPK or induce cytokine production from PMC. Moreover, Spn infection of TNF-α pre-stimulated PMC inhibited production of IL-6 and IL-8 secretion by >50% (p<0.01. In summary, this descriptive study provides datasets and a platform for examining further the molecular mechanisms underlying the pathogenesis of empyema.

  7. Streptococcus pneumoniae Genome-wide Identification and Characterization of BOX Element-binding Domains.

    Zhang, Qiao; Wang, Changzheng; Wan, Min; Wu, Yin; Ma, Qianli

    2015-11-01

    The BOX elements are short repetitive DNA sequences that distribute randomly in intergenic regions of the Streptococcus pneumoniae genome. The function and origin of such elements are still unknown, but they were found to modulate expression of neighboring genes. Evidences suggested that the modulation's mechanism can be fulfilled by sequence-specific interaction of BOX elements with transcription factor family proteins. However, the type and function of these BOX-binding proteins still remain largely unexplored to date. In the current study we described a synthetic protocol to investigate the recognition and interaction between a highly conserved site of BOX elements and the DNA-binding domains of a variety of putative transcription factors in the pneumococcal genome. With the protocol we were able to predict those high-affinity domain binders of the conserved BOX DNA site (BOX DNA) in a high-throughput manner, and analyzed sequence-specific interaction in the domainDNA recognition at molecular level. Consequently, a number of putative transcription factor domains with both high affinity and specificity for the BOX DNA were identified, from which the helix-turn-helix (HTH) motif of a small heat shock factor was selected as a case study and tested for its binding capability toward the double-stranded BOX DNA using fluorescence anisotropy analysis. As might be expected, a relatively high affinity was detected for the interaction of HTH motif with BOX DNA with dissociation constant at nanomolar level. Molecular dynamics simulation, atomic structure examination and binding energy analysis revealed a complicated network of intensive nonbonded interactions across the complex interface, which confers both stability and specificity for the complex architecture. PMID:27491035

  8. Clustering of serotypes in a longitudinal study of Streptococcus pneumoniae carriage in three day care centres

    Tanskanen Antti

    2008-12-01

    Full Text Available Abstract Background Streptococcus pneumoniae (pneumococcus causes a wide range of clinical manifestations that together constitute a major burden of disease worldwide. The main route of pneumococcal transmission is through asymptomatic colonisation of the nasopharynx. Studies of transmission are currently of general interest because of the impact of the new conjugate-polysaccharide vaccines on nasopharyngeal colonisation (carriage. Here we report the first longitudinal study of pneumococcal carriage that records serotype specific exposure to pneumococci simultaneously within the two most important mixing groups, families and day care facilities. Methods We followed attendees (N = 59 with their family members (N = 117 and the employees (N = 37 in three Finnish day care centres for 9 months with monthly sampling of nasopharyngeal carriage. Pneumococci were cultured, identified and serotyped by standard methods. Results Children in day care constitute a core group of pneumococcal carriage: of the 36 acquisitions of carriage with documented exposure to homologous pneumococci, the attendee had been exposed in her/his day care centre in 35 cases and in the family in 9 cases. Day care children introduce pneumococci to the family: 66% of acquisitions of a new serotype in a family were associated with simultaneous or previous carriage of the same type in the child attending day care. Consequently, pneumococcal transmission was found to take place as micro-epidemics driven by the day care centres. Each of the three day care centres was dominated by a serotype of its own, accounting for 100% of the isolates of that serotype among all samples from the day care attendees. Conclusion The transmission of pneumococci is more intense within than across clusters defined by day care facilities. The ensuing micro-epidemic behaviour enhances pneumococcal transmission.

  9. Clustering of serotypes in a longitudinal study of Streptococcus pneumoniae carriage in three day care centres

    Leino, Tuija; Hoti, Fabian; Syrjänen, Ritva; Tanskanen, Antti; Auranen, Kari

    2008-01-01

    Background Streptococcus pneumoniae (pneumococcus) causes a wide range of clinical manifestations that together constitute a major burden of disease worldwide. The main route of pneumococcal transmission is through asymptomatic colonisation of the nasopharynx. Studies of transmission are currently of general interest because of the impact of the new conjugate-polysaccharide vaccines on nasopharyngeal colonisation (carriage). Here we report the first longitudinal study of pneumococcal carriage that records serotype specific exposure to pneumococci simultaneously within the two most important mixing groups, families and day care facilities. Methods We followed attendees (N = 59) with their family members (N = 117) and the employees (N = 37) in three Finnish day care centres for 9 months with monthly sampling of nasopharyngeal carriage. Pneumococci were cultured, identified and serotyped by standard methods. Results Children in day care constitute a core group of pneumococcal carriage: of the 36 acquisitions of carriage with documented exposure to homologous pneumococci, the attendee had been exposed in her/his day care centre in 35 cases and in the family in 9 cases. Day care children introduce pneumococci to the family: 66% of acquisitions of a new serotype in a family were associated with simultaneous or previous carriage of the same type in the child attending day care. Consequently, pneumococcal transmission was found to take place as micro-epidemics driven by the day care centres. Each of the three day care centres was dominated by a serotype of its own, accounting for 100% of the isolates of that serotype among all samples from the day care attendees. Conclusion The transmission of pneumococci is more intense within than across clusters defined by day care facilities. The ensuing micro-epidemic behaviour enhances pneumococcal transmission. PMID:19116005

  10. Diverse Ecological Strategies Are Encoded by Streptococcus pneumoniae Bacteriocin-Like Peptides.

    Miller, Eric L; Abrudan, Monica I; Roberts, Ian S; Rozen, Daniel E

    2016-01-01

    The opportunistic pathogen Streptococcus pneumoniae is commonly carried asymptomatically in the human nasopharynx. Due to high rates of cocolonization with other pneumococcus strains, intraspecific competitive interactions partly determine the carriage duration of strains and thereby their potential to cause disease. These interactions may be mediated by bacteriocins, such as the type IIb bacteriocins encoded by the blp (bacteriocin-like peptide) locus. To understand blp diversity and evolution, we undertook a bioinformatic analysis of 4,418 pneumococcal genomes, including 168 newly sequenced genomes. We describe immense variation at all levels of genomic organization: Gene presence/absence, gene order, and allelic diversity. If we make the extreme and naive hypothesis that assumes all genes in this operon can assort randomly, this variation could lead to 10(15) distinct bacteriocin-related phenotypes, each potentially representing a unique ecological strategy; however, we provide several explanations for why this extreme is not realized. Although rarefaction analysis indicates that the number of unique strategies is not saturated, even after sampling thousands of genomes, we show that the variation is neither unbounded nor random. We delimit three bacteriocin groups, which contain group-specific bacteriocins, immunity genes, and blp operon gene order, and argue that this organization places a constraint on realized ecological strategies. We additionally show that ecological strategy diversity is significantly constrained by pneumococcal phylogeny and clonal structure. By examining patterns of association between alleles within the blp operon, we show that bacteriocin genes, which were believed to function in pairs, can be found with a broad diversity of partner alleles and immunity genes; this overall lack of allelic fidelity likely contributes to the fluid structure of this operon. Our results clarify the diversity of antagonistic ecological strategies in the

  11. Mechanistic investigation of the endo-alpha-N-acetylgalactosaminidase from Streptococcus pneumoniae R6.

    Willis, Lisa M; Zhang, Ran; Reid, Anne; Withers, Stephen G; Wakarchuk, Warren W

    2009-11-01

    The large (1767-amino acid) endo-alpha-N-acetylgalactosaminidase from Streptococcus pneumoniae (SpGH101) specifically removes an O-linked disaccharide Gal-beta-1,3-GalNAc-alpha from glycoproteins. While the enzyme from natural sources has been used as a reagent for many years, very few mechanistic studies have been performed. Using the recently determined three-dimensional structure of the recombinant protein as a background, we report here a mechanistic investigation of the SpGH101 retaining alpha-glycoside hydrolase using a combination of synthetic and natural substrates. On the basis of a model of the substrate complex of SpGH101, we propose D764 and E796 as the nucleophile and general acid-base residues, respectively. These roles were confirmed by kinetic and mechanistic analysis of mutants at those positions using synthetic substrates and anion rescue experiments. pK(a) values of 5.3 and 7.2 were assigned to D764 and E796 on the basis of the pK(a) values derived from the bell-shaped dependence of k(cat)/K(m) upon pH. The enzyme contains several putative carbohydrate binding modules whose glycan binding specificities were probed using the printed glycan array of the Consortium for Functional Glycomics using the inactive D764A and D764F mutants that had been labeled with Alexafluor 488. These studies revealed binding to galacto-N-biose, consistent with a role for these domains in localizing the enzyme near its substrates. PMID:19788271

  12. Identification of surface-exposed proteins and surface-derived structures from Streptococcus pneumoniae for vaccine and diagnostic tools purposes

    Olaya-Abril, Alfonso

    2014-01-01

    Streptococcus pneumoniae (neumococo) es un patógeno humano que puede colonizar el tracto respiratorio superior o puede causar infecciones invasivas como neumonía, meningitis y empiema, y no invasivas como otitis media, sinusitis y bronquitis, principalmente en niños, ancianos e inmunodeprimidos. Es la principal causa de muertes en niños menores de 5 años a nivel mundial, sobre todo en los países en vías de desarrollo. Existen más de 90 serotipos diferentes de acuerdo a la compo...

  13. Heteroduplex DNA mismatch repair system of Streptococcus pneumoniae: cloning and expression of the hexA gene

    Mutations affecting heteroduplex DNA mismatch repair in Streptococcus pneumoniae were localized in two genes, hexA and hexB, by fractionation of restriction fragments carrying mutant alleles. A fragment containing the hexA4 allele was cloned in the S. pneumoniae cloning system, and the hexA+ allele was introduced into the recombinant plasmid by chromosomal facilitation of plasmid transfer. Subcloning localized the functional hexA gene to a 3.5-kilobase segment of the cloned pneumococcal DNA. The product of this gene was shown in Bacillus subtilis minicells to be a polypeptide with an M/sub r/ of 86,000. Two mutant alleles of hexA showed partial expression of the repair system when present in multicopy plasmids. A model for mismatch repair, which depends on the interaction of two protein components to recognize the mismatched base pair and excise a segment of DNA between strand breaks surrounding the mismatch, is proposed

  14. Seasonal Variation of Escherichia coli, Staphylococcus aureus, and Streptococcus pneumoniae Bacteremia According to Acquisition and Patient Characteristics

    Gradel, Kim Oren; Nielsen, Stig Lønberg; Pedersen, Court; Knudsen, Jenny Dahl; Østergaard, Christian; Arpi, Magnus; Jensen, Thøger Gorm; Kolmos, Hans Jørn; Søgaard, Mette; Lassen, Annmarie Touborg; Schønheyder, Henrik Carl

    Seasonal variation analysis. METHODS In 3 Danish health regions (2.3 million total inhabitants), patients with bacteremia were identified from 2000 through 2011 using information from laboratory information systems. Analyses were confined to Escherichia coli, Staphylococcus aureus, and Streptococcus...... pneumoniae. Additional data were obtained from the Danish National Hospital Registry for the construction of admission histories and calculation of the Charlson comorbidity index (CCI). Bacteremias were categorized as community acquired, healthcare associated (HCA), and hospital acquired. We defined multiple....... coli, 6,924 S. aureus, and 4,884 S. pneumoniae bacteremia cases. For E. coli, the seasonal variation was highest for community-acquired cases (PTT ratio, 1.24; 95% CI, 1.17-1.32), was diminished for HCA (PTT ratio, 1.14; 95% CI, 1.04-1.25), and was missing for hospital-acquired cases. No seasonal...

  15. Increased lethality and defective pulmonary clearance of Streptococcus pneumoniae in microsomal prostaglandin E synthase-1-knockout mice.

    Dolan, Jennifer M; Weinberg, Jason B; O'Brien, Edmund; Abashian, Anya; Procario, Megan C; Aronoff, David M; Crofford, Leslie J; Peters-Golden, Marc; Ward, Lindsay; Mancuso, Peter

    2016-06-01

    The production of prostaglandin E2 (PGE2) increases dramatically during pneumococcal pneumonia, and this lipid mediator impairs alveolar macrophage (AM)-mediated innate immune responses. Microsomal prostaglandin E synthase-1 (mPGES-1) is a key enzyme involved in the synthesis of PGE2, and its expression is enhanced during bacterial infections. Genetic deletion of mPGES-1 in mice results in diminished PGE2 production and elevated levels of other prostaglandins after infection. Since PGE2 plays an important immunoregulatory role during bacterial pneumonia we assessed the impact of mPGES-1 deletion in the host defense against pneumococcal pneumonia in vivo and in AMs in vitro. Wild-type (WT) and mPGES-1 knockout (KO) mice were challenged with Streptococcus pneumoniae via the intratracheal route. Compared with WT animals, we observed reduced survival and increased lung and spleen bacterial burdens in mPGES-1 KO mice 24 and 48 h after S. pneumoniae infection. While we found modest differences between WT and mPGES-1 KO mice in pulmonary cytokines, AMs from mPGES-1 KO mice exhibited defective killing of ingested bacteria in vitro that was associated with diminished inducible nitric oxide synthase expression and reduced nitric oxide (NO) synthesis. Treatment of AMs from mPGES-1 KO mice with an NO donor restored bacterial killing in vitro. These results suggest that mPGES-1 plays a critical role in bacterial pneumonia and that genetic ablation of this enzyme results in diminished pulmonary host defense in vivo and in vitro. These results suggest that specific inhibition of PGE2 synthesis by targeting mPGES-1 may weaken host defense against bacterial infections. PMID:27059285

  16. Two Pharmacodynamic Models for Assessing the Efficacy of Amoxicillin-Clavulanate against Experimental Respiratory Tract Infections Caused by Strains of Streptococcus pneumoniae

    Woodnutt, Gary; Berry, Valerie

    1999-01-01

    Two models of respiratory tract infection were used to investigate the pharmacodynamics of amoxicillin-clavulanate against Streptococcus pneumoniae. Eight strains of S. pneumoniae were used in a mouse model in which the animals were infected intranasally and were then treated with a range of doses and dose intervals. The time that the plasma amoxicillin concentration remained above the MIC (T>MIC) correlated well with bacterial killing, such that if T>MIC was below 20% there was no effect on ...

  17. High prevalence of Streptococcus pneumoniae in adenoids and nasopharynx in preschool children with recurrent upper respiratory tract infections in Poland – distribution of serotypes and drug resistance patterns

    Niedzielski, Artur; Korona-Glowniak, Izabela; Malm, Anna

    2013-01-01

    Background Streptococcus pneumoniae is one of the major bacterial pathogens colonizing nasopharynx, and often causes upper respiratory tract infections in children. We investigated the prevalence of S. pneumoniae in nasopharynx and adenoid core in 57 children aged 2–5 years who underwent adenoidectomy for recurrent pharyngotonsillitis, and we determined serotypes and antibiotic resistance patterns of the isolated pneumococci. Material/Methods The nasopharyngeal specimens obtained before adeno...

  18. A novel computational method identifies intra- and inter-species recombination events in Staphylococcus aureus and Streptococcus pneumoniae.

    Lisa Sanguinetti

    Full Text Available Advances in high-throughput DNA sequencing technologies have determined an explosion in the number of sequenced bacterial genomes. Comparative sequence analysis frequently reveals evidences of homologous recombination occurring with different mechanisms and rates in different species, but the large-scale use of computational methods to identify recombination events is hampered by their high computational costs. Here, we propose a new method to identify recombination events in large datasets of whole genome sequences. Using a filtering procedure of the gene conservation profiles of a test genome against a panel of strains, this algorithm identifies sets of contiguous genes acquired by homologous recombination. The locations of the recombination breakpoints are determined using a statistical test that is able to account for the differences in the natural rate of evolution between different genes. The algorithm was tested on a dataset of 75 genomes of Staphylococcus aureus and 50 genomes comprising different streptococcal species, and was able to detect intra-species recombination events in S. aureus and in Streptococcus pneumoniae. Furthermore, we found evidences of an inter-species exchange of genetic material between S. pneumoniae and Streptococcus mitis, a closely related commensal species that colonizes the same ecological niche. The method has been implemented in an R package, Reco, which is freely available from supplementary material, and provides a rapid screening tool to investigate recombination on a genome-wide scale from sequence data.

  19. Fatal purpura fulminans and Waterhouse-Friderichsen syndrome from fulminant Streptococcus pneumoniae sepsis in an asplenic young adult.

    Hale, Andrew J; LaSalvia, Mary; Kirby, James E; Kimball, Allison; Baden, Rachel

    2016-01-01

    Asplenic patients are at increased risk for sepsis and fulminant infection. Sepsis in these patients is typically secondary to encapsulated bacteria, with Streptococcus pneumoniae being the most frequent pathogen. Rare complications of severe sepsis include purpura fulminans and bilateral adrenal hemorrhage (Waterhouse-Friderichsen syndrome). We present the case of a 36-year-old woman, healthy except for splenectomy years prior for idiopathic thrombocytopenic purpura treatment, who presented with fever. Upon presentation to our hospital, three hours after symptoms onset, she had purpura fulminans and shock. Despite timely antimicrobials and maximal resuscitative efforts, her disease progressed and she expired 12 hours after symptoms onset. Autopsy revealed bilateral adrenal hemorrhage; acute adrenal crisis likely contributed to her refractory shock. Prior to her presentation, she had not received guideline-based post-splenectomy care. Sepsis in asplenic patients can be fulminant and rapidly fatal. Streptococcus pneumoniae remains the most frequent cause, despite decreasing rates in recent years related to widespread pneumococcal vaccination. Guideline-based vaccinations and "pill-in-pocket" therapy can be life-saving for asplenic patients. Purpura fulminans represents an extreme manifestation of disseminated intravascular coagulation, is more common in asplenic patients, and portends a poor prognosis. Waterhouse-Friderichsen syndrome can be seen concurrently with purpura fulminans and further portends a poor prognosis; pre-mortem diagnosis requires a high index of suspicion. PMID:27583208

  20. 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.

    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.

  1. Prevalence of antimicrobial resistance of Streptococcus pneumoniae in Chinese children: four hospitals surveillance

    沈叙庄; 陆权; 叶启慈; 张国成; 俞桑洁; 张泓; 邓秋莲; 杨永弘

    2003-01-01

    Objective To investigate the nasal carriage of antibiotic-resistant pneumococci in children of <5 years old in the following four cities, Beijing, Shanghai, Guangzhou and Xi'an.Methods A total of 647 pneumococci strains were isolated and detected. Minimal inhibition concentrations (MICs) of antibiotics were determined by E-test. Disk diffusion test was used for the measurement of antimicrobial susceptibility.Results Prevalence of penicillin non-susceptible Streptococcus pneumoniae in the four cities was 41%, with Guangzhou (60.8%) ranking first, followed by Xi'an (45%), Shanghai (37%) and Beijing (25.9%). The majority of penicillin non-susceptibility isolates (23.9%-53.8%) had a low level of resistance (MIC 0.64-1.5 μg/ml). The most sensitive antimicrobials in terms of percentage of susceptible organisms were amoxicillin-clavulanic acid (99.4%), followed by ceftriaxone (92.1%); cefurxime and cefaclor were slightly more sensitive than penicillin with susceptibility of 74.8% and 77.9%. Erythromycin, tetracycline and TMP-SMZ were highly resistant (83.6%, 82.1% and 76.2% respectively). Among erythromycin resistant isolates, 100% were resistant to azithromycin, 98.6% to clarithromycin, 97.2% to roxithromycin and spiramycin, and 96.6% to clindamycin. 97.2% (141/145) were typical of the macrolides-lincosamides-streptogramons B (MLSB ) resistance phenotype, and 2.8% (4/145) were M phenotype. The group of PRSP was with significantly higher rates of non-susceptibility for ceftriaxone (18.4%), cefurxime (58.6%), cefaclor (53.4%), compared with the group of PEN-S (0.5%, 1.8% and 0.2%, respectively) and the rate of multi-drug resistance in the isolates of PRSP group (92.9%) was significantly higher than that of PEN-S group (59.2%).Conclusion The rates of penicillin and multi-drug resistance among isolates of pneumococci carried nasally in are high children and the high prevalence of multi-drug resistance in the Chinese population may be becoming one of the most serious

  2. Antibiotic selection pressure and macrolide resistance in nasopharyngeal Streptococcus pneumoniae: a cluster-randomized clinical trial.

    Alison H Skalet

    Full Text Available BACKGROUND: It is widely thought that widespread antibiotic use selects for community antibiotic resistance, though this has been difficult to prove in the setting of a community-randomized clinical trial. In this study, we used a randomized clinical trial design to assess whether macrolide resistance was higher in communities treated with mass azithromycin for trachoma, compared to untreated control communities. METHODS AND FINDINGS: In a cluster-randomized trial for trachoma control in Ethiopia, 12 communities were randomized to receive mass azithromycin treatment of children aged 1-10 years at months 0, 3, 6, and 9. Twelve control communities were randomized to receive no antibiotic treatments until the conclusion of the study. Nasopharyngeal swabs were collected from randomly selected children in the treated group at baseline and month 12, and in the control group at month 12. Antibiotic susceptibility testing was performed on Streptococcus pneumoniae isolated from the swabs using Etest strips. In the treated group, the mean prevalence of azithromycin resistance among all monitored children increased from 3.6% (95% confidence interval [CI] 0.8%-8.9% at baseline, to 46.9% (37.5%-57.5% at month 12 (p = 0.003. In control communities, azithromycin resistance was 9.2% (95% CI 6.7%-13.3% at month 12, significantly lower than the treated group (p < 0.0001. Penicillin resistance was identified in 0.8% (95% CI 0%-4.2% of isolates in the control group at 1 year, and in no isolates in the children-treated group at baseline or 1 year. CONCLUSIONS: This cluster-randomized clinical trial demonstrated that compared to untreated control communities, nasopharyngeal pneumococcal resistance to macrolides was significantly higher in communities randomized to intensive azithromycin treatment. Mass azithromycin distributions were given more frequently than currently recommended by the World Health Organization's trachoma program. Azithromycin use in this setting

  3. Kinetic and Binding Studies of Streptococcus pneumoniae Type 2 Isopentenyl Diphosphate:Dimethylallyl Diphosphate Isomerase.

    Janczak, Matthew Walter; Poulter, C Dale

    2016-04-19

    Type 2 isopentenyl diphosphate:dimethylallyl diphosphate isomerase (IDI-2) converts isopentenyl diphosphate (IPP) to dimethylallyl diphosphate (DMAPP), the two fundamental building blocks of isoprenoid molecules. IDI-2 is found in many species of bacteria and is a potential antibacterial target since this isoform is non-homologous to the type 1 enzyme in Homo sapiens. IDI-2 requires a reduced flavin mononucleotide to form the catalytically active ternary complex, IDI-2·FMNH2·IPP. For IDI-2 from the pathogenic bacterium Streptococcus pneumoniae, the flavin can be treated kinetically as a dissociable cosubstrate in incubations with IPP and excess NADH. Under these conditions, the enzyme follows a modified sequential ordered mechanism where FMN adds before IPP. Interestingly, the enzyme shows sigmoidal behavior when incubated with IPP and NADH with varied concentrations of FMN in aerobic conditions. In contrast, sigmoidal behavior is not seen in incubations under anaerobic conditions where FMN is reduced to FMNH2 before the reaction is initiated by addition of IPP. Stopped-flow experiments revealed that FMN, whether bound to IDI-2 or without enzyme in solution, is slowly reduced in a pseudo-first-order reaction upon addition of excess NADH (kred(FMN) = 5.7 × 10(-3) s(-1) and kred(IDI-2·FMN) = 2.8 × 10(-3) s(-1)), while reduction of the flavin is rapid upon addition of NADH to a mixture of IDI-2·FMN, and IPP (kred(IDI-2·FMN·IPP) = 8.9 s(-1)). Similar experiments with dithionite as the reductant gave kred(FMN) = 221 s(-1) and kred(IDI-2·FMN) = 411 s(-1). Dithionite reduction of FMN in the IDI-2·FMN and IPP mixture was biphasic with kred(IDI-2·FMN·IPP (fast)) = 326 s(-1) and kred(IDI-2·FMN·IPP (slow)) = 6.9 s(-1) The pseudo-first-order rate constant for the slow component was similar to those for NADH reduction of the flavin in the IDI-2·FMN and IPP mixture and may reflect a rate-limiting conformational change in the enzyme. PMID:27003727

  4. Factors Contributing to Hydrogen Peroxide Resistance in Streptococcus pneumoniae Include Pyruvate Oxidase (SpxB) and Avoidance of the Toxic Effects of the Fenton Reaction

    Pericone, Christopher D.; Park, Sunny; Imlay, James A.; Weiser, Jeffrey N.

    2003-01-01

    Aerobic growth of Streptococcus pneumoniae results in production of amounts of hydrogen peroxide (H2O2) that may exceed 1 mM in the surrounding media. H2O2 production by S. pneumoniae has been shown to kill or inhibit the growth of other respiratory tract flora, as well as to have cytotoxic effects on host cells and tissue. The mechanisms allowing S. pneumoniae, a catalase-deficient species, to survive endogenously generated concentrations of H2O2 that are sufficient to kill other bacterial s...

  5. Children from 0 to 4 Years Old Carrying Streptococcus Pneumoniae. A Community Study Niños de 0 a 4 años portadores de Streptococcus pneumoniae. Estudio en la comunidad

    Olga Olivia Tejeda Hernández

    2011-08-01

    Full Text Available Background: Streptococcus pneumoniae is the leading cause of community-acquired pneumonia. The infection is preceded by nasopharyngeal colonization. Therefore, it becomes very important to determine the carrier’s health condition as well as patterns of susceptibility and resistance. Objective: To determine the prevalence of the Streptococcus pneumoniae carrier state among a population of children under four years old. Methods: A cross-sectional study including 179 children from urban and rural areas in San Antonio de los Baños, Artemisa was conducted. The municipality was stratified by the number of births in each year, the percentage they represented in the sample was determined and a number of children (proportional to the size of the stratified group was placed in each group. Children were randomly selected in different clinics. Samples from nasal and pharyngeal swabs were collected. Strains identification was performed by morphological and cultural characteristics, the optochin test and the bile solubility. Swab cultures confirmation was performed by latex agglutination. Antimicrobial susceptibility was determined by the Kirby-Bauer method. Results: 20, 6% of children were nasal carriers and 33, 6% were pharyngeal carriers, the last ones being more frequent in urban areas. There were no differences by sex; 60% of the strains were susceptible to penicillin. A 52% of resistance to trimethoprim-sulfamethoxazole and a 25% to erythromycin was found. The total number of isolated strains was susceptible to ofloxacin, cefotaxime, azithromycin, and vancomycin. Conclusions: The carrier state was high.Fundamento: el Streptococcus pneumoniae es la primera causa de neumonías comunitarias; a la infección la precede la colonización de la nasofaringe, de ahí la importancia de determinar el estado de portador y los patrones de susceptibilidad y

  6. Ascorbic acid-dependent gene expression in Streptococcus pneumoniae and the activator function of the transcriptional regulator UlaR2

    Afzal, Muhammad; Shafeeq, Sulman; Kuipers, Oscar P

    2015-01-01

    In this study, we have explored the impact of ascorbic acid on the transcriptome of Streptococcus pneumoniae D39. The expression of several genes and operons, including the ula operon (which has been previously shown to be involved in ascorbic acid utilization), the AdcR regulon (which has been prev

  7. Cellobiose-Mediated Gene Expression in Streptococcus pneumoniae : A Repressor Function of the Novel GntR-Type Regulator BguR

    Shafeeq, Sulman; Kuipers, Oscar P.; Kloosterman, Tomas G.; Leite, Luciana C.C.

    2013-01-01

    The human pathogen Streptococcus pneumoniae has the ability to use the carbon-and energy source cellobiose due to the presence of a cellobiose-utilizing gene cluster (cel locus) in its genome. This system is regulated by the cellobiose-dependent transcriptional activator CelR, which has been previou

  8. Clinical application of a ligation-independent pathway of multiplex ligation-dependent probe amplification for the determination of quinolone susceptibility of Streptococcus pneumoniae.

    Uno, Naoki; Araki, Nobuko; Kaku, Norihito; Kosai, Kosuke; Hasegawa, Hiroo; Yanagihara, Katsunori

    2016-09-01

    We previously uncovered a ligation-independent pathway of multiplex ligation-dependent probe amplification (MLPA) through which products of MLPA could be amplified without both hybridization and ligation reactions. Here, we utilized this pathway to detect an antibiotic resistance mutation of quinolones in Streptococcus pneumoniae. PMID:27343683

  9. New Real-Time PCR Assay Using Locked Nucleic Acid Probes To Assess Prevalence of ParC Mutations in Fluoroquinolone-Susceptible Streptococcus pneumoniae Isolates from France

    Decousser, Jean-Winoc; Methlouthi, Imen; Pina, Patrick; Collignon, Anne; Allouch, Pierre

    2006-01-01

    A real-time PCR assay with locked nucleic acid probes was developed to screen mutations at codons 79 and 83 of the Streptococcus pneumoniae parC gene. Only silent mutations were detected among 236 French invasive fluoroquinolone-susceptible strains. This test could be useful for some high-risk patients or in national surveys.

  10. New real-time PCR assay using locked nucleic acid probes to assess prevalence of ParC mutations in fluoroquinolone-susceptible Streptococcus pneumoniae isolates from France.

    Decousser, Jean-Winoc; Methlouthi, Imen; Pina, Patrick; Collignon, Anne; Allouch, Pierre

    2006-04-01

    A real-time PCR assay with locked nucleic acid probes was developed to screen mutations at codons 79 and 83 of the Streptococcus pneumoniae parC gene. Only silent mutations were detected among 236 French invasive fluoroquinolone-susceptible strains. This test could be useful for some high-risk patients or in national surveys. PMID:16569894

  11. Pneumonia

    ... viruses, such as the influenza virus (flu) and adenovirus . Other viruses, such as respiratory syncytial virus (RSV) ... your local health department to see when these vaccines are available. Because pneumonia ... influenza pneumonia, for example, someone may become sick as ...

  12. Serotype 19A bacteremic pneumococcal pneumonia after 4 doses of 13-valent conjugate vaccine: a review of pneumococcal conjugate vaccine effectiveness

    IrohTam, Pui-Ying; Hanisch, Benjamin R.; Forward, Brennan

    2014-01-01

    We report a case of bacteremic pneumococcal pneumonia due to serotype 19A in a child who had received four doses of the 13-valent pneumococcal conjugate vaccine, and review the literature on effectiveness of this vaccine. Pediatricians should be alert to the fact that up-to-date immunization status with pneumococcal vaccine does not preclude illness from invasive disease caused by a vaccine serotype.

  13. A genomic approach to understand interactions between Streptococcus pneumoniae and its bacteriophages

    Leprohon, Philippe; Gingras, Hélène; Ouennane, Siham; Moineau, Sylvain; Ouellette, Marc

    2015-01-01

    Background Bacteriophage replication depends on bacterial proteins and inactivation of genes coding for such host factors should interfere with phage infection. To gain further insights into the interactions between S. pneumoniae and its pneumophages, we characterized S. pneumoniae mutants selected for resistance to the virulent phages SOCP or Dp-1. Results S. pneumoniae R6-SOCPR and R6-DP1R were highly resistant to the phage used for their selection and no cross-resistance between the two ph...

  14. Alcohol abuse and Streptococcus pneumoniae infections: Consideration of Virulence Factors and Impaired Immune Responses

    Bhatty, Minny; Pruett, Stephen B.; Swiatlo, Edwin; Nanduri, Bindu

    2011-01-01

    Alcohol is the most frequently abused substance in the world. Both acute and chronic alcohol consumption have diverse and well documented effects on the human immune system, leading to increased susceptibility to infections like bacterial pneumonia. S. pneumoniae is the most common bacterial etiology of community acquired pneumonia world-wide. The frequency and severity of pneumococcal infections in individuals with a history of alcohol abuse is much higher than the general population. Despit...

  15. Gene expression in the DpnI and DpnII restriction enzyme systems of Streptococcus pneumoniae

    Lacks, S.A.; Sabelnikov, A.G.; Chen, Jau-Der; Greenberg, B.

    1992-12-31

    Although a number of bacterial species are naturally transformable, that is, their cells are able to take up external DNA in substantial amounts and integrate it into the chromosome without artificial manipulation of the cell surface, Streptococcus pneumoniae, the first species in which this phenomenon was detected, remains a prototype of such transformation. Cells of S. pneumonias also contain potent restriction endonucleases able to severely restrict DNA introduced during viral infection. Our current understanding of the genetic basis of the complementary DpnI and DpnII restriction systems and of the biochemistry of their component enzymes are briefly reviewed. The manner in which these enzymes impinge on the transfer of chromosomal genes and of plasmeds will be examined in detail. It will be seen that far from acting against foreign DNA in general, the restriction systems seem to be designed to exclude only infecting viral DNA The presence of complementary restriction systems in different cells of S. pneumonias enhances their effectiveness in blocking viral infection and promoting species survival. This enhanced effectiveness requires the expression of alternative restriction systems. Therefore, the ability of the cells to transfer the restriction enzyme genes and to regulate their expression are important for survival of the species.

  16. Meningitis caused by Neisseria Meningitidis, Hemophilus Influenzae Type B and Streptococcus Pneumoniae during 2005–2012 in Turkey

    Ceyhan, Mehmet; Gürler, Nezahat; Ozsurekci, Yasemin; Keser, Melike; Aycan, Ahmet Emre; Gurbuz, Venhar; Salman, Nuran; Camcioglu, Yildiz; Dinleyici, Ener Cagri; Ozkan, Sengul; Sensoy, Gulnar; Belet, Nursen; Alhan, Emre; Hacimustafaoglu, Mustafa; Celebi, Solmaz; Uzun, Hakan; Faik Oner, Ahmet; Kurugol, Zafer; Ali Tas, Mehmet; Aygun, Denizmen; Oncel, Eda Karadag; Celik, Melda; Yasa, Olcay; Akin, Fatih; Coşkun, Yavuz

    2014-01-01

    Successful vaccination policies for protection from bacterial meningitis are dependent on determination of the etiology of bacterial meningitis. Cerebrospinal fluid (CSF) samples were obtained prospectively from children from 1 month to ≤ 18 years of age hospitalized with suspected meningitis, in order to determine the etiology of meningitis in Turkey. DNA evidence of Neisseria meningitidis (N. meningitidis), Streptococcus pneumoniae (S. pneumoniae), and Hemophilus influenzae type b (Hib) was detected using multiplex polymerase chain reaction (PCR). In total, 1452 CSF samples were evaluated and bacterial etiology was determined in 645 (44.4%) cases between 2005 and 2012; N. meningitidis was detected in 333 (51.6%), S. pneumoniae in 195 (30.2%), and Hib in 117 (18.1%) of the PCR positive samples. Of the 333 N. meningitidis positive samples 127 (38.1%) were identified as serogroup W-135, 87 (26.1%) serogroup B, 28 (8.4%) serogroup A and 3 (0.9%) serogroup Y; 88 (26.4%) were non-groupable. As vaccines against the most frequent bacterial isolates in this study are available and licensed, these results highlight the need for broad based protection against meningococcal disease in Turkey. PMID:25483487

  17. Cloning, recombinant production, crystallization and preliminary X-ray diffraction analysis of a family 101 glycoside hydrolase from Streptococcus pneumoniae

    The catalytic module of a family 101 glycoside hydrolase from S. pneumoniae was cloned, recombinantly produced and crystallized. Streptococcus pneumoniae is a serious human pathogen that is responsible for a wide range of diseases including pneumonia, meningitis, septicaemia and otitis media. The full virulence of this bacterium is reliant on carbohydrate processing and metabolism, as revealed by biochemical and genetic studies. One carbohydrate-processing enzyme is a family 101 glycoside hydrolase (SpGH101) that is responsible for catalyzing the liberation of galactosyl β1,3-N-acetyl-d-galactosamine (Galβ1,3GalNAc) α-linked to serine or threonine residues of mucin-type glycoproteins. The 124 kDa catalytic module of this enzyme (SpGH101CM) was cloned and overproduced in Escherichia coli and purified. Crystals were obtained in space group P21 and diffracted to 2.0 Å resolution, with unit-cell parameters a = 81.86, b = 88.91, c = 88.77 Å, β = 112.46°. SpGH101CM also qualitatively displayed good activity towards the synthetic substrate p-nitrophenyl-2-acetamido-2-deoxy-3-O-(β-d-galactopyranosyl) -α-d-galactopyranoside, which is consistent with the classification of this enzyme as an endo-α-N-acetylgalactosaminidase

  18. A case report of bacteremia manifesting as an overwhelming postsplenectomy infection due to Streptococcus pneumoniae post vaccination.

    Hirose, Kosuke; Okabe, Hirohisa; Yoshizumi, Tomoharu; Uchiyama, Hideaki; Ikegami, Toru; Harimoto, Norifumi; Itoh, Shinji; Kimura, Koichi; Baba, Hideo; Maehara, Yoshihiko

    2016-12-01

    A 62-year-old woman was admitted for acute epigastralgia and high-grade fever of over 39 °C. The patient had undergone splenectomy for idiopathic portal hypertension 1 year ago and vaccination against Streptococcus pneumoniae immediately post operation. She developed localized peritoneal irritation and abdominal distension. Her serum creatinine had increased to 1.5 mg/dL and procalcitonin was 12.5 ng/ml. Computed tomography of the abdomen revealed edematous large intestine and increased ascites. From these results, the patient was considered to have spontaneous bacterial peritonitis (SBP). Vancomycin (VCM) and doripenem (DRPM) were administered to control the infection. Unexpectedly, S. pneumoniae was detected in the blood culture. Hence, ampicillin/sulbactam was administered after discontinuing VCM. The patient recovered without any life-threatening complications and was discharged after 10 days. In conclusion, overwhelming postsplenectomy infection (OPSI) due to S. pneumoniae could develop in patient with splenectomy even after vaccination. Although the bacteremia probably due to SBP and acute renal dysfunction was accompanied by OPSI, our patient recovered rapidly. PMID:27221131

  19. An explosive outbreak of Streptococcus pneumoniae serotype-8 infection in a highly vaccinated residential care home, England, summer 2012.

    Thomas, H L; Gajraj, R; Slack, M P E; Sheppard, C; Hawkey, P; Gossain, S; Drew, C M; Pebody, R G

    2015-07-01

    In August 2012, an explosive outbreak of severe lower respiratory tract infection (LRTI) due to Streptococcus pneumoniae serotype-8 occurred in a highly vaccinated elderly institutionalized population in England. Fifteen of 23 residents developed LRTI over 4 days (attack rate 65%); 11 had confirmed S. pneumoniae serotype-8 disease, and two died. Following amoxicillin chemoprophylaxis and pneumococcal polysaccharide vaccine (PPV) re-vaccination no further cases occurred in the following 2 months. No association was found between being an outbreak-associated case and age (P = 0.36), underlying comorbidities [relative risk (RR) 0.84 95% confidence interval (CI) 0.34-2.09], or prior receipt of PPV (RR 1.4, 95% CI 0.60-3.33). However, the median number of years since PPV was significantly higher for cases (n = 15, 10.2 years, range 7.3-17.9 years) than non-cases (n = 8, 7.2 years, range 6.8-12.8 years) (P = 0.045), provided evidence of waning immunity. Alternative vaccination strategies should be considered to prevent future S. pneumoniae outbreaks in institutionalized elderly populations. PMID:25298247

  20. Co2+-dependent gene expression in Streptococcus pneumoniae: opposite effect of Mn2+ and Co2+ on the expression of the virulence genes psaBCA, pcpA, and prtA

    Manzoor, Irfan; Shafeeq, Sulman; Kloosterman, Tomas G.; Oscar P. Kuipers

    2015-01-01

    Manganese (Mn2+)-, zinc (Zn2+)- and copper (Cu2+) play significant roles in transcriptional gene regulation, physiology, and virulence of Streptococcus pneumoniae. So far, the effect of the important transition metal ion cobalt (Co2+) on gene expression of S. pneumoniae has not yet been explored. Here, we study the impact of Co2+ stress on the transcriptome of S. pneumoniae strain D39. BLAST searches revealed that the genome of S. pneumoniae encodes a putative Co2+-transport operon (cbi opero...

  1. Co2+-dependent gene expression in Streptococcus pneumoniae: Opposite effect of Mn2+ and Co2+ on the expression of the virulence genes psaBCA, pcpA and prtA

    Irfan eManzoor; Sulman eShafeeq; Kloosterman, Tomas G.; Oscar P. Kuipers

    2015-01-01

    Manganese (Mn2+)-, zinc (Zn2+)- and copper (Cu2+) play significant roles in transcriptional gene regulation, physiology and virulence of Streptococcus pneumoniae. So far, the effect of the important transition metal ion cobalt (Co2+) on gene expression of S. pneumoniae has not yet been explored. Here, we study the impact of Co2+ stress on the transcriptome of S. pneumoniae strain D39. BLAST searches revealed that the genome of S. pneumoniae encodes a putative Co2+-transport operon (cbi operon...

  2. Allergic lung inflammation alters neither susceptibility to Streptococcus pneumoniae infection nor inducibility of innate resistance in mice

    Evans Christopher M

    2009-07-01

    Full Text Available Abstract Background Protective host responses to respiratory pathogens are typically characterized by inflammation. However, lung inflammation is not always protective and it may even become deleterious to the host. We have recently reported substantial protection against Streptococcus pneumoniae (pneumococcal pneumonia by induction of a robust inflammatory innate immune response to an inhaled bacterial lysate. Conversely, the allergic inflammation associated with asthma has been proposed to promote susceptibility to pneumococcal disease. This study sought to determine whether preexisting allergic lung inflammation influences the progression of pneumococcal pneumonia or reduces the inducibilty of protective innate immunity against bacteria. Methods To compare the effect of different inflammatory and secretory stimuli on defense against pneumonia, intraperitoneally ovalbumin-sensitized mice were challenged with inhaled pneumococci following exposure to various inhaled combinations of ovalbumin, ATP, and/or a bacterial lysate. Thus, allergic inflammation, mucin degranulation and/or stimulated innate resistance were induced prior to the infectious challenge. Pathogen killing was evaluated by assessing bacterial CFUs of lung homogenates immediately after infection, the inflammatory response to the different conditions was evaluated by measurement of cell counts of bronchoalveolar lavage fluid 18 hours after challenge, and mouse survival was assessed after seven days. Results We found no differences in survival of mice with and without allergic inflammation, nor did the induction of mucin degranulation alter survival. As we have found previously, mice treated with the bacterial lysate demonstrated substantially increased survival at seven days, and this was not altered by the presence of allergic inflammation or mucin degranulation. Allergic inflammation was associated with predominantly eosinophilic infiltration, whereas the lysate-induced response

  3. Association of Streptococcus pneumoniae nasopharyngeal colonization and other risk factors with acute otitis media in an unvaccinated Indian birth cohort.

    Rupa, V; Isaac, R; Rebekah, G; Manoharan, A

    2016-07-01

    In order to study the epidemiology of acute otitis media (AOM) and Streptococcus pneumoniae nasopharyngeal colonization in the first 2 years of life, we followed up an unvaccinated birth cohort monthly and at visits when sick, with otoscopy to detect AOM and performed nasopharyngeal swabbing to detect S. pneumoniae. Serotyping of positive cultures was also performed. Of 210 babies who were enrolled at birth, 61 (29·05%) experienced 128 episodes of AOM [relative risk 2·63, 95% confidence interval (CI) 1·21-5·75] with maximum incidence in the second half of the first year of life. Episodes ranged from 1 to 7 (mean 2·1 episodes). Most (86·9%) babies with AOM had a positive culture swab giving an odds ratio (OR) of 1·93 (95% CI 1·03-3·62, P = 0·041) for this association. Other risk factors identified for AOM were winter season (OR 3·46, 95% CI 1·56-7·30, P = 0·001), upper respiratory infection (OR 2·43, 95% CI 1·43-4·51, P = 0·005); residents of small households were less likely to develop AOM (OR 0·32, 95% CI 0·17-0·57, P < 0·01). Common S. pneumoniae serotypes isolated during episodes were 19, 6, 15, 35, 7, 23, 9 and 10 which indicated a theoretical coverage for pneumococcal vaccines PCV10 and PCV13 constituent serotypes of 62·8%. We conclude that AOM in Indian infants is often associated with S. pneumoniae colonization of the nasopharynx as well as other risk factors. PMID:26931207

  4. Characterization of biofilm matrix, degradation by DNase treatment and evidence of capsule downregulation in Streptococcus pneumoniae clinical isolates

    Johnson Candice

    2008-10-01

    Full Text Available Abstract Background Streptococcus pneumoniae is a common respiratory pathogen and a major causative agent of respiratory infections, including otitis media (OM. Pneumococcal biofilms have been demonstrated on biopsies of the middle ear mucosa in children receiving tympanostomy tubes, supporting the hypothesis that chronic OM may involve biofilm development by pathogenic bacteria as part of the infectious process. To better understand pneumococcal biofilm formation six low-passage encapsulated nasopharyngeal isolates of S. pneumoniae were assessed over a six-eight day period in vitro. Results Multiparametric analysis divided the strains into two groups. Those with a high biofilm forming index (BFI were structurally complex, exhibited greater lectin colocalization and were more resistant to azithromycin. Those with a low BFI developed less extensive biofilms and were more susceptible to azithromycin. dsDNA was present in the S. pneumoniae biofilm matrix in all strains and treatment with DNase I significantly reduced biofilm biomass. Since capsule expression has been hypothesized to be associated with decreased biofilm development, we also examined expression of cpsA, the first gene in the pneumococcal capsule operon. Interestingly, cpsA was downregulated in biofilms in both high and low BFI strains. Conclusion All pneumococcal strains developed biofilms that exhibited extracellular dsDNA in the biofilm matrix, however strains with a high BFI correlated with greater carbohydrate-associated structural complexity and antibiotic resistance. Furthermore, all strains of S. pneumoniae showed downregulation of the cpsA gene during biofilm growth compared to planktonic culture, regardless of BFI ranking, suggesting downregulation of capsule expression occurs generally during adherent growth.

  5. Rapid detection of Streptococcus pneumoniae by real-time fluorescence quantitation PCR%实时荧光定量PCR快速诊断肺炎链球菌

    颜善活; 孙雷; 符可鹏; 卓永光; 杨善业; 樊祖茜; 黄永霞

    2013-01-01

    目的 建立实时荧光定量聚合酶链反应(PCR),用于肺炎链球菌检测和流行病学调查.方法 以肺炎链球菌自溶素(lyt)和溶血素(ply)的基因为目的序列分别设计引物和探针,将其分别与10株肺炎链球菌、13株非肺炎链球菌DNA以及不同浓度梯度的肺炎链球菌DNA进行荧光定量PCR,并将引物和探针应用于200例临床标本检测,同时通过传统的微生物培养鉴定的方法来验证荧光定量PCR的特异性和敏感性.结果 10株肺炎链球菌均获得了明显的扩增产物,13株非肺炎链球菌DNA无明显的扩增信号,其检测敏感性可达100 fg;200例临床标本,实时荧光定量PCR检测出42例肺炎链球菌阳性(阳性率为21%),而培养法阳性16例(阳性率为8%).结论 实时荧光定量PCR是一种敏感、特异、快速的检测肺炎链球菌方法,可用于肺炎链球菌的诊断和流行病学调查.%Objective To establish real-time fluorescence quantitation polymerase chain reaction ( PCR) for the detection and epidemiological studies of Streptococcus pneumoniae. Methods The autolysin (lyt) gene and hemolysin (ply) gene sequences were selected to develop primers and probe. A total of 10 strains of Streptococcus pneumoniae,13 strains of non-Streptococcus pneumoniae DISA and the different concentration strains of Streptococcus pneumoniae DNA were detected by real-time fluorescence quantitation PCR, and 200 clinical specimens were detected by primers and probe. The specificity and sensitivity were also analyzed. Results The 10 strains of Streptococcus pneumoniae were measured to obtain amplification products. However, 13 strains of non-Streptococcus pneumoniae DNA had no obvious amplification, and the sensitivity was 100fg. Among the 200 clinical specimens, real-time fluorescence quantitation PCR detected 42 cases of Streptococcus pneumoniae-positive (the positive rate was 21% ), while the culture method detected 16 cases of Streptococcus pneumoniae

  6. Susceptibility of respiratory isolates of Streptococcus pneumoniae isolated from children hospitalized in the Clinical center Niš

    Dinić Marina M.

    2016-01-01

    Full Text Available Introduction. Streptococcus pneumoniae is one of the most common causes of respiratory infections. The aim was to study the susceptibility to antimicrobial agents of respiratory isolates of Streptococcus pneumoniae obtained from hospitalized children. Material and Methods. A total of 190 respiratory pneumococcal isolates obtained from children aged from 0 to 14 years were isolated and identified by using standard microbiological methods. Susceptibility to oxacillin, erythromycin, clindamycin, tetracycline, cotrimoxazole, ofloxacin and rifampicin was tested by disc diffusion method. Minimal inhibitory concentrations for amoxicillin and ceftriaxone were determined by means of E test. The macrolide-resistant phenotype was detected by double disc diffusion test. Results. All tested isolates were susceptible to amoxicillin and ceftriaxone. The minimal amoxicillin concentration inhibiting the growth of 50% of isolates and of 90% of isolates was 0.50 μg/ml and 1.0 μg/ml, respectively and the minimal ceftriaxone concentration inhibiting the growth of 50% of isolates and of 90% of isolates was 0.25 μg/ml and 0.50 μg/ml, respectively. Susceptibility to erythromycin and clindamycin was observed in 21.6% and 29.47% of isolates, respectively. The resistence to macrolides - M phenotype was detected in 10.07% of isolates and constitutive macrolide-lincosamide-streptogramin phenotype (constitutive MLS phenotype was found in 89.93% of isolates. All tested isolates were susceptible to ofloxacin and rifampicin. Conclusion. Amoxicillin could be the therapy of choice in pediatric practice. The macrolides should not be recommended for the empirical therapy of pneumococcal respiratory tract infection in our local area.

  7. Treatment of Streptococcus pneumoniae disease in children%儿童肺炎链球菌性疾病的治疗

    陆权

    2011-01-01

    肺炎链球菌性疾病(尤其侵袭性肺炎链球菌性疾病)是5岁以下儿童较常见的感染性疾病,肺炎链球菌对抗菌药物的耐药性给临床治疗带来新的挑战.本文综述儿科肺炎链球菌性疾病、肺炎链球菌的耐药现状,并重点评论肺炎链球菌性疾病的治疗策略,包括肺炎链球菌性肺炎、中耳炎、鼻窦炎、脑膜炎和其他侵袭性肺炎链球菌性疾病,积极防治肺炎链球菌性疾病将加速联合国千年发展目标的实现.%Streptococcus pneumoniae disease(PD) especially as invasive pneumococcal disease (IPD)is more common in children younger than 5. The resistance of Streptococcus pneumoniae against antibacterial agents brings new challenges for clinical treatment. This paper reviews PD and Streptococcus pneumoniae resistance, especially focuses on the strategies of treatment for PD including pneumococcal pneumonia, otitis media, sinusitis, meningitis and other invasive pneumococcal diseases. Active prevention and control of pneumococcal disease will speed up the implementation of the United Nations Millennium Development Goals.

  8. A study on molecular characterization of macrolide resistance mechanism among isolates of Streptococcus pneumoniae from the southern Marmara region of Turkey, as well as resistance to macrolides and penicillin in these isolates

    ÖZAKIN, Cüneyt; GÜLER, Hicran; GÜRCÜOĞLU, Emel; ÖZBEY, Saliha BAKIR; Kazak, Esra; SINIRTAŞ, Ayşe Melda

    2012-01-01

    To determine the distribution of macrolide resistance genes as well as resistance rates in isolated Streptococcus pneumoniae strains from the southern Marmara region of Turkey. Materials and methods: Antimicrobial resistance rates and MIC values were determined by the E-test method in 300 Streptococcus pneumoniae isolates that were isolated from clinical samples. Resistance genes were determined by the polymerase chain reaction (PCR) method in erythromycin-resistant strains. Results: It ...

  9. ANALYSIS OF STREPTOCOCCUS PNEUMONIA 'S DRUG RESISTANCE TO MACROLIDES%肺炎链球菌对大环内酯类耐药性分析

    郁宝慧

    2015-01-01

    目的 探索郯城县区肺炎链球菌分离株对大环内酯类耐药性及其治疗情况.方法 收集郯城县社区和临床分离的肺炎链球菌 ,根据CLSI标准,用K-B法、Etest法检测肺炎链球菌对红霉素和克林霉素的耐药性及对红霉素的耐药表型.结果 493株肺炎链球菌中红霉素不敏感菌株489株 ,对克林霉素不敏感菌株482株,其中 ermB基因介导的红霉素耐药菌株482株 ,占98.6% , mefE基因介导的红霉素耐药菌株7株,占1.4%.结论 郯城县区对红霉素和克林霉素近乎全部耐药 ,ermB基因介导的靶位改变是郯城县区肺炎链球菌对大环内酯类抗菌药物的主要耐药机制.%Objective To explore streptococcus pneumonia's drug resistance to macrolides . Methods Samples of streptococcus pneumonia were collected from the communities in Tancheng County and the iso-lated ones from clinical treatment .According to the CLSI standard ,K-B method and Etest were used to detect streptococcus pneumonia ' s drug resistance to erythromycin and clindamycin and the resistance phenotype of erythromycin . Results In the 493 strains of streptococcus pneumonia ,489 strains were not sensi-tive to erythromycin ,482 strains were not sensitive to clindamycin .482 drug-resistant strains were mediated by ermB gene ,accounting for 98 .6% of all the strains .7 drug-resistant strains were mediated by mefE gene ,accounting for 1 .4% .Conclusion Erythro-mycin and clindamycin has no obvious effect on streptococcus pneumoniae in this county . T he target change of mediating by ErmB gene is the main mechanism of streptococcus pneumonia 's resistance to mac-rolides .

  10. Choline Binding Proteins from Streptococcus pneumoniae: A Dual Role as Enzybiotics and Targets for the Design of New Antimicrobials.

    Maestro, Beatriz; Sanz, Jesús M

    2016-01-01

    Streptococcus pneumoniae (pneumococcus) is an important pathogen responsible for acute invasive and non-invasive infections such as meningitis, sepsis and otitis media, being the major cause of community-acquired pneumonia. The fight against pneumococcus is currently hampered both by insufficient vaccine coverage and by rising antimicrobial resistances to traditional antibiotics, making necessary the research on novel targets. Choline binding proteins (CBPs) are a family of polypeptides found in pneumococcus and related species, as well as in some of their associated bacteriophages. They are characterized by a structural organization in two modules: a functional module (FM), and a choline-binding module (CBM) that anchors the protein to the choline residues present in the cell wall through non-covalent interactions. Pneumococcal CBPs include cell wall hydrolases, adhesins and other virulence factors, all playing relevant physiological roles for bacterial viability and virulence. Moreover, many pneumococcal phages also make use of hydrolytic CBPs to fulfill their infectivity cycle. Consequently, CBPs may play a dual role for the development of novel antipneumococcal drugs, both as targets for inhibitors of their binding to the cell wall and as active cell lytic agents (enzybiotics). In this article, we review the current state of knowledge about host- and phage-encoded pneumococcal CBPs, with a special focus on structural issues, together with their perspectives for effective anti-infectious treatments. PMID:27314398

  11. Crystallization and preliminary crystallographic analysis of the transpeptidase domain of penicillin-binding protein 2B from Streptococcus pneumoniae

    Yamada, Mototsugu, E-mail: mototsugu-yamada@meiji.co.jp; Watanabe, Takashi; Baba, Nobuyoshi; Miyara, Takako; Saito, Jun; Takeuchi, Yasuo [Pharmaceutical Research Center, Meiji Seika Kaisha Ltd, 760 Morooka-cho, Kohoku-ku, Yokohama 222-8567 (Japan)

    2008-04-01

    The selenomethionyl-substituted transpeptidase domain of penicillin-binding protein (PBP) 2B from S. pneumoniae was isolated from a limited proteolysis digest of the soluble form of recombinant PBP 2B and then crystallized. MAD data were collected to 2.4 Å resolution. Penicillin-binding protein (PBP) 2B from Streptococcus pneumoniae catalyzes the cross-linking of peptidoglycan precursors that occurs during bacterial cell-wall biosynthesis. A selenomethionyl (SeMet) substituted PBP 2B transpeptidase domain was isolated from a limited proteolysis digest of a soluble form of recombinant PBP 2B and then crystallized. The crystals belonged to space group P4{sub 3}2{sub 1}2, with unit-cell parameters a = b = 86.39, c = 143.27 Å. Diffraction data were collected to 2.4 Å resolution using the BL32B2 beamline at SPring-8. The asymmetric unit contains one protein molecule and 63.7% solvent.

  12. Crystallization and preliminary X-ray analysis of enoyl-acyl carrier protein reductase (FabK) from Streptococcus pneumoniae

    Saito, Jun, E-mail: jun-saito@meiji.co.jp; Yamada, Mototsugu; Watanabe, Takashi; Kitagawa, Hideo; Takeuchi, Yasuo [Pharmaceutical Research Center, Meiji Seika Kaisha Ltd, 760 Morooka-cho, Kohoku-ku, Yokohama 222-8567 (Japan)

    2006-06-01

    Enoyl-acyl carrier protein (ACP) reductases are responsible for bacterial type II fatty-acid biosynthesis and are attractive targets for developing novel antibiotics. The S. pneumoniae enoyl-ACP reductase (FabK) was crystallized and selenomethionine MAD data were collected to 2 Å resolution. The enoyl-acyl carrier protein (ACP) reductase from Streptococcus pneumoniae (FabK; EC 1.3.1.9) is responsible for catalyzing the final step in each elongation cycle of fatty-acid biosynthesis. Selenomethionine-substituted FabK was purified and crystallized by the hanging-drop vapour-diffusion method at 277 K. The crystal belongs to space group P2{sub 1}, with unit-cell parameters a = 50.26, b = 126.70, c = 53.63 Å, β = 112.46°. Diffraction data were collected to 2.00 Å resolution using synchrotron beamline BL32B2 at SPring-8. Two molecules were estimated to be present in the asymmetric unit, with a solvent content of 45.1%.

  13. The interactome of Streptococcus pneumoniae and its bacteriophages show highly specific patterns of interactions among bacteria and their phages.

    Mariano, Rachelle; Wuchty, Stefan; Vizoso-Pinto, Maria G; Häuser, Roman; Uetz, Peter

    2016-01-01

    Although an abundance of bacteriophages exists, little is known about interactions between their proteins and those of their bacterial hosts. Here, we experimentally determined the phage-host interactomes of the phages Dp-1 and Cp-1 and their underlying protein interaction network in the host Streptococcus pneumoniae. We compared our results to the interaction patterns of E. coli phages lambda and T7. Dp-1 and Cp-1 target highly connected host proteins, occupy central network positions, and reach many protein clusters through the interactions of their targets. In turn, lambda and T7 targets cluster to conserved and essential proteins in E. coli, while such patterns were largely absent in S. pneumoniae. Furthermore, targets in E. coli were mutually strongly intertwined, while targets of Dp-1 and Cp-1 were strongly connected through essential and orthologous proteins in their immediate network vicinity. In both phage-host systems, the impact of phages on their protein targets appears to extend from their network neighbors, since proteins that interact with phage targets were located in central network positions, have a strong topologically disruptive effect and touch complexes with high functional heterogeneity. Such observations suggest that the phages, biological impact is accomplished through a surprisingly limited topological reach of their targets. PMID:27103053

  14. Multicentric study in five African countries of antibiotic susceptibility for three main pathogens: Streptococcus pneumoniae, Staphylococcus aureus, and Pseudomonas aeruginosa.

    Zerouali, Khalid; Ramdani-Bouguessa, Nadjia; Boye, Cheikh; Hammami, Adnane

    2016-08-01

    Antibiotic resistance is a growing clinical and epidemiological problem. We report on the antibiotic susceptibility of three pathogens isolated from patients in Algeria, Egypt, Morocco, Senegal, and Tunisia during 2010-2011. In total, 218 Streptococcus pneumoniae, 428 Staphylococcus aureus, and 414 Pseudomonas aeruginosa strains were collected. S. pneumoniae resistance was noted against penicillin (30.2%), erythromycin (27.4%), cefpodoxime (19.1%), amoxicillin (12.0%), cefotaxime (7.4%), and levofloxacin (3.2%). All the strains were teicoplanin susceptible. Staphylococcus aureus methicillin resistance differed between countries, from 5.0% in Senegal to 62.7% in Egypt. Levofloxacin resistance was low in all countries, and the highest rate (in Egypt) was still only 13.6% for intermediate and resistant strains combined. Most strains were susceptible to fosfomycin (99.3%) and pristinamycin (94.2%). P. aeruginosa resistance was found against levofloxacin (30.4%), ciprofloxacin (29.9%), tobramycin (19.7%), ceftazidime (19.2%), and imipenem (17.9%), but not colistin. Antibiotic susceptibility varied widely between countries, with resistance typically most prevalent in Egypt. PMID:25363146

  15. Cloning, recombinant production, crystallization and preliminary X-ray diffraction analysis of a family 101 glycoside hydrolase from Streptococcus pneumoniae.

    Gregg, Katie J; Boraston, Alisdair B

    2009-02-01

    Streptococcus pneumoniae is a serious human pathogen that is responsible for a wide range of diseases including pneumonia, meningitis, septicaemia and otitis media. The full virulence of this bacterium is reliant on carbohydrate processing and metabolism, as revealed by biochemical and genetic studies. One carbohydrate-processing enzyme is a family 101 glycoside hydrolase (SpGH101) that is responsible for catalyzing the liberation of galactosyl beta1,3-N-acetyl-D-galactosamine (Galbeta1,3GalNAc) alpha-linked to serine or threonine residues of mucin-type glycoproteins. The 124 kDa catalytic module of this enzyme (SpGH101CM) was cloned and overproduced in Escherichia coli and purified. Crystals were obtained in space group P2(1) and diffracted to 2.0 A resolution, with unit-cell parameters a = 81.86, b = 88.91, c = 88.77 A, beta = 112.46 degrees. SpGH101CM also qualitatively displayed good activity towards the synthetic substrate p-nitrophenyl-2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)-alpha-D-galactopyranoside, which is consistent with the classification of this enzyme as an endo-alpha-N-acetylgalactosaminidase. PMID:19194003

  16. The interactome of Streptococcus pneumoniae and its bacteriophages show highly specific patterns of interactions among bacteria and their phages

    Mariano, Rachelle; Wuchty, Stefan; Vizoso-Pinto, Maria G.; Häuser, Roman; Uetz, Peter

    2016-01-01

    Although an abundance of bacteriophages exists, little is known about interactions between their proteins and those of their bacterial hosts. Here, we experimentally determined the phage-host interactomes of the phages Dp-1 and Cp-1 and their underlying protein interaction network in the host Streptococcus pneumoniae. We compared our results to the interaction patterns of E. coli phages lambda and T7. Dp-1 and Cp-1 target highly connected host proteins, occupy central network positions, and reach many protein clusters through the interactions of their targets. In turn, lambda and T7 targets cluster to conserved and essential proteins in E. coli, while such patterns were largely absent in S. pneumoniae. Furthermore, targets in E. coli were mutually strongly intertwined, while targets of Dp-1 and Cp-1 were strongly connected through essential and orthologous proteins in their immediate network vicinity. In both phage-host systems, the impact of phages on their protein targets appears to extend from their network neighbors, since proteins that interact with phage targets were located in central network positions, have a strong topologically disruptive effect and touch complexes with high functional heterogeneity. Such observations suggest that the phages, biological impact is accomplished through a surprisingly limited topological reach of their targets. PMID:27103053

  17. Dual Acting Neuraminidase Inhibitors Open New Opportunities to Disrupt the Lethal Synergism between Streptococcus pneumoniae and Influenza Virus

    Elisabeth eWalther

    2016-03-01

    Full Text Available Secondary infections with Streptococcus pneumoniae cause severe pneumonia and enhance lethality during influenza epidemics and pandemics. Structural and functional similarities with viral neuraminidase (NA suggest that the highly prevalent pneumococcal NAs, NanA and NanB, might contribute to this lethal synergism by supporting viral replication and that dual acting NA inhibitors (NAIs will disrupt it. To verify this hypothesis, NanA and NanB were expressed in E. coli. After confirming their activity in enzyme assays, in vitro models with influenza virus A/Jena/8178/09 (Jena/8178 and the recombinant NanA or NanB (rNanA and rNanB were established in A549 and MDCK cells to mimic the role of these pneumococcal NAs during co-infection. Studies on the influence of both NAs on viral receptor expression, spread, and yield revealed a distinct effect of NanA and NanB on viral replication in these in vitro models. Both enzymes were able to support Jena/8178 replication at certain concentrations. This synergism was disrupted by the NAIs oseltamivir, DANA, katsumadain A, and artocarpin exerting an inhibitory effect on viral NA and NanA. Interestingly, katsumadain A and artocarpin inhibited rNanA and rNanB similarly. Zanamivir did not show activity. These results demonstrate a key role of pneumococcal NAs in the lethal synergism with influenza viruses and reveal opportunities for its effective disruption.

  18. Role of an iron-dependent transcriptional regulator in the pathogenesis and host response to infection with Streptococcus pneumoniae.

    Radha Gupta

    Full Text Available Iron is a critical cofactor for many enzymes and is known to regulate gene expression in many bacterial pathogens. Streptococcus pneumoniae normally inhabits the upper respiratory mucosa but can also invade and replicate in lungs and blood. These anatomic sites vary considerably in both the quantity and form of available iron. The genome of serotype 4 pneumococcal strain TIGR4 encodes a putative iron-dependent transcriptional regulator (IDTR. A mutant deleted at idtr (Δidtr exhibited growth kinetics similar to parent strain TIGR4 in vitro and in mouse blood for up to 48 hours following infection. However, Δidtr was significantly attenuated in a murine model of sepsis. IDTR down-regulates the expression of ten characterized and putative virulence genes in nasopharyngeal colonization and pneumonia. The host cytokine response was significantly suppressed in sepsis with Δidtr. Since an exaggerated inflammatory response is associated with a poor prognosis in sepsis, the decreased inflammatory response could explain the increased survival with Δidtr. Our results suggest that IDTR, which is dispensable for pneumococcal growth in vitro, is associated with regulation of pneumococcal virulence in specific host environments. Additionally, IDTR ultimately modulates the host cytokine response and systemic inflammation that contributes to morbidity and mortality of invasive pneumococcal disease.

  19. In vitro biofilm development of Streptococcus pneumoniae and formation of choline-binding protein-DNA complexes.

    Domenech, Mirian; Ruiz, Susana; Moscoso, Miriam; García, Ernesto

    2015-10-01

    Extracellular deoxyribonucleic acid (eDNA) is an essential component of bacterial biofilm matrices, and is required in their formation and maintenance. Extracellular DNA binds to exopolysaccharides or extracellular proteins, affording biofilms greater structural integrity. Recently, we reported evidence of intercellular eDNA-LytC complexes in pneumococcal biofilms. The LytC lysozyme is a member of the choline-binding family of proteins (CBPs) located on the pneumococcal surface. The present work shows that other CBPs, i.e. LytA, LytB, Pce, PspC and CbpF, which have a pI between 5 and 6, can bind DNA in vitro. This process requires the presence of divalent cations other than Mg(2+). This DNA binding capacity of CBPs appears to be independent of their enzymatic activity and, at least in the case of LytA, does not require the choline-binding domain characteristic of CBPs. Positively charged, surface-exposed, 25 amino acid-long peptides derived from the catalytic domain of LytB, were also found capable of DNA binding through electrostatic interactions. Confocal laser scanning microcopy revealed the existence of cell-associated LytB-eDNA complexes in Streptococcus pneumoniae biofilms. These and other findings suggest that these surface-located proteins of S. pneumoniae could play roles of varying importance in the colonization and/or invasion of human host where different environmental conditions exist. PMID:25950767

  20. Pneumonia

    ... restroom and before eating. Use lukewarm water and soap for at least 20 seconds. If soap and water are not available, using an alcohol- ... at higher risk for pneumonia? Do I have bacterial, viral or fungal pneumonia? What’s the best treatment? ...

  1. Trends in antibacterial resistance among Streptococcus pneumoniae isolated in the USA: update from PROTEKT US Years 1–4

    Brown Steven D

    2008-01-01

    Full Text Available Abstract Background The increasing prevalence of resistance to established antibiotics among key bacterial respiratory tract pathogens, such as Streptococcus pneumoniae, is a major healthcare problem in the USA. The PROTEKT US study is a longitudinal surveillance study designed to monitor the susceptibility of key respiratory tract pathogens in the USA to a range of commonly used antimicrobials. Here, we assess the geographic and temporal trends in antibacterial resistance of S. pneumoniae isolates from patients with community-acquired respiratory tract infections collected between Year 1 (2000–2001 and Year 4 (2003–2004 of PROTEKT US. Methods Antibacterial minimum inhibitory concentrations were determined centrally using the Clinical and Laboratory Standards Institute (CLSI broth microdilution method; susceptibility was defined according to CLSI interpretive criteria. Macrolide resistance genotypes were determined by polymerase chain reaction. Results A total of 39,495 S. pneumoniae isolates were collected during 2000–2004. The percentage of isolates resistant to erythromycin, penicillin, levofloxacin, and telithromycin were 29.3%, 21.2%, 0.9%, and 0.02%, respectively, over the 4 years, with marked regional variability. The proportion of isolates exhibiting multidrug resistance (includes isolates known as penicillin-resistant S. pneumoniae and isolates resistant to ≥ 2 of the following antibiotics: penicillin; second-generation cephalosporins, e.g. cefuroxime; macrolides; tetracyclines; and trimethoprim-sulfamethoxazole remained stable at ~30% over the study period. Overall mef(A was the most common macrolide resistance mechanism. The proportion of mef(A isolates decreased from 68.8% to 62.3% between Year 1 and Year 4, while the percentage of isolates carrying both erm(B and mef(A increased from 9.7% to 18.4%. Over 99% of the erm(B+mef(A-positive isolates collected over Years 1–4 exhibited multidrug resistance. Higher than previously

  2. Streptococcus pneumoniae infection regulates expression of neurotrophic factors in the olfactory bulb and cultured olfactory ensheathing cells.

    Ruiz-Mendoza, S; Macedo-Ramos, H; Santos, F A; Quadros-de-Souza, L C; Paiva, M M; Pinto, T C A; Teixeira, L M; Baetas-da-Cruz, W

    2016-03-11

    Streptococcus pneumoniae is the causative agent of numerous diseases including severe invasive infections such as bacteremia and meningitis. It has been previously shown that strains of S. pneumoniae that are unable to survive in the bloodstream may colonize the CNS. However, information on cellular components and pathways involved in the neurotropism of these strains is still scarce. The olfactory system is a specialized tissue in which olfactory receptor neurons (ORNs) are interfacing with the external environment through several microvilli. Olfactory ensheathing cells (OECs) which also form the glial limiting membrane at the surface of the olfactory bulb (OB) are the only cells that ensheathe the ORNs axons. Since previous data from our group showed that OECs may harbor S. pneumoniae, we decided to test whether infection of the OB or OEC cultures modulates the expression levels of neurotrophic factor's mRNA and its putative effects on the activation and viability of microglia. We observed that neurotrophin-3 (NT-3) and glial cell-line-derived neurotrophic factor (GDNF) expression was significantly higher in the OB from uninfected mice than in infected mice. A similar result was observed when we infected OEC cultures. Brain-derived neurotrophic factor (BNDF) expression was significantly lower in the OB from infected mice than in uninfected mice. In contrast, in vitro infection of OECs resulted in a significant increase of BDNF mRNA expression. An upregulation of high-mobility group box 1 (HMGB1) expression was observed in both OB and OEC cultures infected with S. pneumoniae. Moreover, we found that conditioned medium from infected OEC cultures induced the expression of the pro-apoptotic protein cleaved-caspase-3 and an apparently continuous nuclear factor-kappa B (NF-κB) p65 activation in the N13 microglia. Altogether, our data suggest the possible existence of an OEC-pathogen molecular interface, through which the OECs could interfere on the activation and

  3. Multiple-locus variable-number tandem-repeat analysis of Streptococcus pneumoniae and comparison with multiple loci sequence typing

    van Cuyck Hélène

    2012-10-01

    Full Text Available Abstract Background Streptococcus pneumoniae infections remain a major cause of morbidity and mortality worldwide. The diversity of pneumococci was first evidenced by serotyping of their capsular polysaccharides, responsible of virulence, resolving into more than 93 serotypes. Molecular tools have been developed to track the emergence and the spread of resistant, hyper virulent or non-vaccine type clones, particularly DNA-based methods using genetic polymorphism. Pulsed-Field Gel Electrophoresis analysis (PFGE and Multiple Loci Sequence Typing (MLST are the most frequently used genotyping techniques for S. pneumoniae. MLST is based on sequence comparison of housekeeping genes clustering isolates within sequence types. The availability of genome sequence data from different S. pneumoniae strains facilitated the search for other class of genetic markers as polymorphic DNA sequences for a Multiple-Locus Variable-Number Tandem-Repeat Analysis (MLVA. This study aims at confirming the relevance of MLVA of S. pneumoniae, comparing MLST and MLVA performances when discriminating subgroups of strains belonging to the same Sequence Type (ST, and defining a restricted but universal set of MLVA markers that has at least the same discriminatory power as MLST for S. pneumoniae by applying marker sets used by different authors on 331 isolates selected in UK. Results A minimum spanning tree was built including the serotypes distribution and comparing MLVA and MLST results. 220 MLVA types were determined grouped in 10 Sequence Types (ST. MLVA differentiated ST162 in two clonal complexes. A minimal set was defined: ms 25 and ms37, ms17, ms19, ms33, ms39, and ms40 including two universal markers. The selection was based on MLVA markers with a Diversity Index >0.8 and a selection of others depending of the population tested and the aim of the study. This set of 7 MLVA markers yields strain clusters similar to those obtained by MLST. Conclusions MLVA can discriminate

  4. Pesquisa de módulos de genes associados à invasividade de Streptococcus pneumoniae usando semelhanças semânticas

    Malaquias, João José dos Reis

    2014-01-01

    Tese de mestrado em Bioquímica, apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2014 O Streptococcus pneumoniae, ou pneumococcus, e uma bactéria causadora de doenças em humanos, tais como pneumonia, meningite bacteriana e otite, entre outras. Apresenta uma elevada taxa de mortalidade em certas faixas etárias e regiões do mundo. Desde a década de 70 do séc. XX que têm sido desenvolvidas vacinas que conseguiram diminuir a taxa de infecção pelo pneumococcus. No entanto...

  5. Characterization of a Novel Fucose-Regulated Promoter (PfcsK) Suitable for Gene Essentiality and Antibacterial Mode-of-Action Studies in Streptococcus pneumoniae

    Chan, Pan F.; O'Dwyer, Karen M.; Palmer, Leslie M.; Ambrad, Jennifer D.; Ingraham, Karen A.; So, Chi; Lonetto, Michael A.; Biswas, Sanjoy; Rosenberg, Martin; Holmes, David J.; Zalacain, Magdalena

    2003-01-01

    The promoter of the Streptococcus pneumoniae putative fuculose kinase gene (fcsK), the first gene of a novel fucose utilization operon, is induced by fucose and repressed by glucose or sucrose. When the streptococcal polypeptide deformylase (PDF) gene (def1, encoding PDF) was placed under the control of PfcsK, fucose-dependent growth of the S. pneumoniae (PfcsK::def1) strain was observed, confirming the essential nature of PDF in this organism. The mode of antibacterial action of actinonin, a...

  6. 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

    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...... 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...

  7. 96株儿童肺炎链球菌耐药情况分析%Resistance of 96 strains of streptococcus pneumoniae in children

    樊有; 姜静; 蒋德升; 尚宁; 施毅

    2011-01-01

    Objective To investigate antibiotic resistance of streptococcus pneumoniae to children in Nanjing.Methods Totally 96 strains of streptococcus pneumoniae were collected to test the MICs of various antibiotics by agar dilution method according to the approved standard of CLIS.Results Among 96 strains of streptococcus pneumoniae ,63 (65.6%) strains were resistant to penicillin ( MIC≥2 mg/L).87.5% ,29.2%,8.3% and 4.2% of streptococcus pneumoniae were resistant against the cefuroxime,cefotaxim,amoxicillin ,and ceftriaxone ,retrospectively.The resistance rates to other antibiotic agents, such as erythromycin, azithromycin,tetracycline,and Chloram-phenicol were 96.9% ,95.8% ,95.8% ,94.8%, respectively.All 96 strains of streptococcus pneumoniae were sensitive to vancomycin, tigocycline and linezolid.Conclusion The antibiotic resistance to streptococcus pneumoniae is serious in Nanjing.Most of them are multi-resistant strains.Except for vancomycin,tigecycline,linezolid, and ceftriaxone, most antibiotic agents have lost the reactivities against streptococcus pneumoniae.%目的 了解南京地区儿童感染肺炎链球菌对常用抗菌药物的耐药性.方法 琼脂稀释法测定96株肺炎链球菌对14种抗菌药物最低抑菌浓度.结果 96株肺炎链球菌中,耐青霉素肺炎链球菌(penicillin resist streptococcus pneumoniae,PRSP)最小抑菌浓度(minimal inhibitory concentration,MIC)≥2mg/L的检出率为65.6%;头孢呋辛、头孢噻肟、阿莫西林、头孢曲松的耐药率依次为87.5%、29.2%、8.3%和4.2%;红霉素、四环素、阿奇霉素和克林霉素耐药率分别为96.9%、95.8%、95.8%、94.8%;万古霉素、替加环素、利奈唑胺均敏感.结论 南京地区儿童肺炎链球菌对青霉素、红霉素、阿奇霉素、克林霉素和四环素、头孢呋辛等抗生素耐药性高,应注意合理选择用药.

  8. Co2+-dependent gene expression in Streptococcus pneumoniae: opposite effect of Mn2+ and Co2+ on the expression of the virulence genes psaBCA, pcpA, and prtA

    Manzoor, Irfan; Shafeeq, Sulman; Kloosterman, Tomas; Kuipers, Oscar

    2015-01-01

    Manganese (Mn2+)-, zinc (Zn2+)- and copper (Cu2+) play significant roles in transcriptional gene regulation, physiology, and virulence of Streptococcus pneumoniae. So far, the effect of the important transition metal ion cobalt (Co2+) on gene expression of S. pneumoniae has not yet been explored. He

  9. Crystal structure of enoyl–acyl carrier protein reductase (FabK) from Streptococcus pneumoniae reveals the binding mode of an inhibitor

    Saito, Jun; Yamada, Mototsugu; Watanabe, Takashi; Iida, Maiko; Kitagawa, Hideo; Takahata, Sho; Ozawa, Tomohiro; Takeuchi, Yasuo; Ohsawa, Fukuichi

    2008-01-01

    Enoyl–acyl carrier protein (ACP) reductases are critical for bacterial type II fatty acid biosynthesis and thus are attractive targets for developing novel antibiotics. We determined the crystal structure of enoyl–ACP reductase (FabK) from Streptococcus pneumoniae at 1.7 Å resolution. There was one dimer per asymmetric unit. Each subunit formed a triose phosphate isomerase (TIM) barrel structure, and flavin mononucleotide (FMN) was bound as a cofactor in the active site. The overall structure...

  10. Streptococcus pneumoniae colonisation in children and adolescents with asthma: impact of the heptavalent pneumococcal conjugate vaccine and evaluation of potential effect of thirteen-valent pneumococcal conjugate vaccine

    Esposito, S.; L. Terranova; M.F. Patria; Marseglia, G L; Miraglia del Giudice, M; Bodini, A; Martelli, A.; Baraldi, E; O. Mazzina; Tagliabue, C.; A. Licari; V. Ierardi; M. Lelii; Principi, N

    2016-01-01

    Background The main aim of this study was to evaluate Streptococcus pneumoniae carriage in a group of school-aged children and adolescents with asthma because these results might indicate the theoretical risk of invasive pneumococcal disease (IPD) of such patients and the potential protective efficacy of the 13-valent pneumococcal conjugate vaccine (PCV13). Methods Oropharyngeal samples were obtained from 423 children with documented asthma (300 males, 70.9 %), and tested for the autolysin-A-...

  11. Alterations of Penicillin-Binding Proteins 1A, 2X, and 2B in Streptococcus pneumoniae Isolates for Which Amoxicillin MICs Are Higher than Penicillin MICs

    Kosowska, K.; Jacobs, M R; Bajaksouzian, S.; Koeth, L.; Appelbaum, P. C.

    2004-01-01

    Penicillin-binding proteins (PBPs) of 15 selected penicillin- and amoxicillin-resistant Streptococcus pneumoniae isolates (MICs of 2 to 8 and 8 to 16 μg/ml, respectively) were studied. In addition to typical changes in PBPs 1A and 2X, these strains had 10 unique changes in PBP 2B, including a 618A-G substitution, which may be the key alteration associated with amoxicillin resistance.

  12. In vitro selection of one-step mutants of Streptococcus pneumoniae resistant to different oral beta-lactam antibiotics is associated with alterations of PBP2x.

    Sifaoui, F; Kitzis, M D; Gutmann, L

    1996-01-01

    Many oral penicillins and cephalosporins are used to treat clinical infections caused by Streptococcus pneumoniae. Therefore, using different beta-lactams as selectors, we estimated the frequencies of one-step mutations leading to resistance. Resistant mutants were obtained from penicillin-susceptible, intermediately resistant, and penicillin resistant strains. For cefixime, cefuroxime, cefpodoxime, cefotaxime, and ceftriaxone, the frequencies of mutation ranged from 10(-6) to 10(-8) when res...

  13. Outbreaks of Streptococcus pneumoniae carriage in day care cohorts in Finland – implications for elimination of transmission

    Auranen Kari; Leino Tuija; Erästö Panu; Hoti Fabian

    2009-01-01

    Abstract Background Day care centre (DCC) attendees play a central role in maintaining the circulation of Streptococcus pneumoniae (pneumococcus) in the population. Exposure within families and within DCCs are the main risk factors for colonisation with pneumococcal serotypes in DCC attendees. Methods Transmission of serotype specific carriage was analysed with a continuous time event history model, based on longitudinal data from day care attendees and their family members. Rates of acquisit...

  14. Drug resistance current situation and mechanism of Streptococcus pneumoniae%肺炎链球菌的耐药现状及其机制研究进展

    景文莹

    2014-01-01

    Streptococcus pneumoniae is one of the main pathogenic bacteria causing community-ac-quired infections in children, is a common pathogenic bacteria of respiratory tract infections, and is a main pathogenic bacteria of meningitis, bacteremia, otitis media and sinusitis. At pres-ent, antibiotics are still the effective way of treatment for pneumococcal disease. But recently, drug-resistance streptococcus pneumoniae to various antibiotics is increasing, and it has a large regional differences in different areas. Multi-drug resistant Streptococcus pneumoniae has become a global problem, this paper reviewed the drug resistance streptococcus pneumoniae and the resis-tance mechanisms to macrolide , tetracycline and quinolone antibiotic.%肺炎链球菌是引起儿童社区获得性感染的主要病原菌之一,是呼吸道感染的常见病原菌,也是脑膜炎、菌血症、中耳炎和鼻窦炎的主要致病菌。目前,抗菌药物的应用仍然是治疗肺炎链球菌疾病的有效手段。然而近年来,世界各地肺炎链球菌对各种抗生素的耐药率不断增加,且不同地区差异较大。肺炎链球菌多重耐药已逐渐成为全球性问题,本文综述了肺炎链球菌的耐药现状及其对大环内酯类、四环素类及喹诺酮类等抗生素的耐药机制。

  15. Type 3-specific synthase of Streptococcus pneumoniae (Cap3B) directs type 3 polysaccharide biosynthesis in Escherichia coli and in pneumococcal strains of different serotypes

    Arrecubieta, C; López, Rubens; García, Ernesto

    1996-01-01

    The cap3B gene, which is involved in the formation of the capsule of Streptococcus pneumoniae type 3, encodes a 49-kD protein that has been identified as a polysaccharide synthase. Escherichia coli cells harboring the recombinant plasmid pTBP3 (cap3B) produced pneumococcal type 3 polysaccharide, as demonstrated by immunological tests. Biochemical and cell fractionation analyses revealed that this polysaccharide had a high molecular mass and was localized in substantial amounts in the periplas...

  16. Distribution and Genetic Diversity of the ABC Transporter Lipoproteins PiuA and PiaA within Streptococcus pneumoniae and Related Streptococci

    Whalan, Rachael H.; Funnell, Simon G.P.; Bowler, Lucas D.; Hudson, Michael J.; Robinson, Andrew; Dowson, Christopher G.

    2006-01-01

    Streptococcus pneumoniae is a major cause of morbidity and mortality worldwide. The existence of approximately 90 antigenically distinct capsular serotypes has greatly complicated the development of an effective pneumococcal vaccine. Virulence-associated proteins common and conserved among all capsular types now represent the best strategy to combat pneumococcal infections. PiuA and PiaA are the lipoprotein components of two pneumococcal iron ABC transporters and are required for full virulen...

  17. In Vitro Activities of Cethromycin (ABT-773), a New Ketolide, against Streptococcus pneumoniae Strains That Are Not Susceptible to Penicillin or Macrolides

    Mason, Edward O.; Lamberth, Linda B.; Wald, Ellen R.; Bradley, John S.; Barson, William J.; Kaplan, Sheldon L.

    2003-01-01

    Pneumococcal resistance to antimicrobials presents problems to physicians for empirical treatment of acute otitis media (AOM). Three hundred thirty-three isolates of Streptococcus pneumoniae selected for nonsusceptibility to penicillin (MIC >0.1 μg/ml) from the middle ear (n = 325) or mastoid (n = 8) of children seen between 1994 and 2000 at four children's hospitals in the United States were tested by broth microdilution for susceptibility to nine antibiotics. Using NCCLS 2002 breakpoints, r...

  18. Development of approaches to a third-generation carbohydrate-conjugate vaccine against Streptococcus pneumoniae: the search for optimal oligosaccharide ligands

    Gening, M. L.; Kurbatova, E. A.; Tsvetkov, Yu E.; Nifantiev, N. E.

    2015-11-01

    The review addresses the application of synthetic oligosaccharides related to fragments of capsular polysaccharides from different serotypes of the bacterium Streptococcus pneumoniae for the design of third-generation pneumococcal conjugate vaccines. Special focus is given to characteristic features of the chemical structures of oligosaccharides required for the induction of the protective immune response when using synthetic glycoconjugate vaccines based on oligosaccharide ligands and carrier proteins. The bibliography includes 101 references.

  19. Population structure and drug resistance patterns of emerging non-PCV-13 Streptococcus pneumoniae serotypes 22F, 15A, and 8 isolated from adults in Ontario, Canada.

    Duvvuri, Venkata R; Deng, Xianding; Teatero, Sarah; Memari, Nader; Athey, Taryn; Fittipaldi, Nahuel; Gubbay, Jonathan B

    2016-08-01

    The introduction of pneumococcal conjugate vaccines has led to the emergence of non-vaccine serotypes, which contributed to invasive pneumococcal disease in Canada and worldwide. A significant increase in the prevalence of non-13-valent pneumococcal conjugate vaccine (PCV-13)-included serotypes 22F, 15A, and 8 was observed from 2009 to 2013 in Ontario (all p valuesStreptococcus pneumoniae population structures and dynamics, and its utility in molecular surveillance. PMID:27071529

  20. PCR-Based Serotyping of Streptococcus pneumoniae from Culture-Negative Specimens: Novel Primers for Detection of Serotypes within Serogroup 18.

    Tanmoy, Arif M; Saha, Senjuti; Darmstadt, Gary L; Whitney, Cynthia G; Saha, Samir K

    2016-08-01

    Six multiplex-compatible PCR primers were designed to distinguish Streptococcus pneumoniae serotypes within serogroup 18 from culturable/nonculturable pneumococcal specimens, with no cross-reactivity with other serotypes and respiratory organisms. These primers will aid in the generation of better data on vaccine/nonvaccine serotypes in invasive and carriage pneumococcal surveillance and contribute to future vaccine formulation and impact studies. PMID:27252464

  1. Fucose-Mediated Transcriptional Activation of the fcs Operon by FcsR in Streptococcus pneumoniae

    Manzoor, Irfan; Shafeeq, Sulman; Afzal, Muhammad; Kuipers, Oscar P

    2015-01-01

    In this study, we explore the impact of fucose on the transcriptome of S. pneumoniae D39. The expression of various genes and operons, including the fucose uptake PTS and utilization operon (fcs operon) was altered in the presence of fucose. By means of quantitative RT-PCR and β-galactosidase analys

  2. Structure of the fucose mutarotase from Streptococcus pneumoniae in complex with l-­fucose

    Melanie A Higgins; Boraston, Alisdair B.

    2011-01-01

    SpFcsU is a fucose mutarotase from S. pneumoniae that links the harvesting of fucose from glycans by glycoside hydrolases to processing of the fucose monosaccharide by subsequent enzymes in a fucose-utilization pathway. Here, the decameric structure of SpFcsU in complex with fucose is described.

  3. From knock-out phenotype to three-dimensional structure of a promising antibiotic target from Streptococcus pneumoniae.

    Con Dogovski

    Full Text Available Given the rise in drug-resistant Streptococcus pneumoniae, there is an urgent need to discover new antimicrobials targeting this pathogen and an equally urgent need to characterize new drug targets. A promising antibiotic target is dihydrodipicolinate synthase (DHDPS, which catalyzes the rate-limiting step in lysine biosynthesis. In this study, we firstly show by gene knock out studies that S. pneumoniae (sp lacking the DHDPS gene is unable to grow unless supplemented with lysine-rich media. We subsequently set out to characterize the structure, function and stability of the enzyme drug target. Our studies show that sp-DHDPS is folded and active with a k(cat = 22 s(-1, K(M(PYR = 2.55 ± 0.05 mM and K(M(ASA = 0.044 ± 0.003 mM. Thermal denaturation experiments demonstrate sp-DHDPS exhibits an apparent melting temperature (T(M(app of 72 °C, which is significantly greater than Escherichia coli DHDPS (Ec-DHDPS (T(M(app = 59 °C. Sedimentation studies show that sp-DHDPS exists in a dimer-tetramer equilibrium with a K(D(4→2 = 1.7 nM, which is considerably tighter than its E. coli ortholog (K(D(4→2 = 76 nM. To further characterize the structure of the enzyme and probe its enhanced stability, we solved the high resolution (1.9 Å crystal structure of sp-DHDPS (PDB ID 3VFL. The enzyme is tetrameric in the crystal state, consistent with biophysical measurements in solution. Although the sp-DHDPS and Ec-DHDPS active sites are almost identical, the tetramerization interface of the s. pneumoniae enzyme is significantly different in composition and has greater buried surface area (800 Å(2 compared to its E. coli counterpart (500 Å(2. This larger interface area is consistent with our solution studies demonstrating that sp-DHDPS is considerably more thermally and thermodynamically stable than Ec-DHDPS. Our study describe for the first time the knock-out phenotype, solution properties, stability and crystal structure of DHDPS from S. pneumoniae, a

  4. SURFACE PROTEINS AND PNEUMOLYSIN OF ENCAPSULATED AND NONENCAPSULATED STREPTOCOCCUS PNEUMONIAE MEDIATE VIRULENCE IN A CHINCHILLA MODEL OF OTITIS MEDIA

    Lance E. Keller

    2016-05-01

    Full Text Available Streptococcus pneumoniae infections result in a range of human diseases and are responsible for almost one million deaths annually. Pneumococcal disease is mediated in part through surface structures and an anti-phagocytic capsule. Recent studies have shown that nonencapsulated Streptococcus pneumoniae (NESp make up a significant portion of the pneumococcal population and are able to cause disease. NESp lack some common surface proteins expressed by encapsulated pneumococci, but express surface proteins unique to NESp. A chinchilla model of otitis media (OM was used to determine the effect various pneumococcal mutations have on pathogenesis in both NESp and encapsulated pneumococci. Epithelial cell adhesion and invasion assays were used to examine the effects in relation to deletion of intrinsic genes or expression of novel genes. A mouse model of colonization was also utilized for comparison of various pneumococcal mutants. It was determined that pneumococcal surface protein K (PspK and pneumolysin (Ply affect NESp middle ear pathogenesis, but only PspK affected epithelial cell adhesion. Experiments in an OM model were done with encapsulated strains testing the importance of native virulence factors and treatment of OM. First, a triple deletion of the common virulence factors PspA, PspC, and Ply, (ΔPAC, from an encapsulated background abolished virulence in an OM model while a PspC mutant had detectable, but reduced amounts of recoverable bacteria compared to wildtype. Next, treatment of OM was effective when starting antibiotic treatment within 24 hrs with resolution by 48 hrs post treatment. Expression of NESp-specific virulence factor PspK in an encapsulated strain has not been previously studied, and we showed significantly increased adhesion and invasion of human epithelial cells by pneumococci. Murine colonization was not significantly increased when an encapsulated strain expressed PspK, but colonization was increased when a capsule

  5. Antibiotic treatment and the diagnosis of Streptococcus pneumoniae in lower respiratory tract infections in adults

    Korsgaard, Jens; Møller, Jens Kjølseth; Kilian, Mogens

    2005-01-01

    patient was positive in both systems, making a total of 22 patients with documented pneumococcal infection. As a positive culture test was dependent on the absence of antibiotic treatment, whereas a positive urine antigen test depended on antibiotic treatment within 48 hours, the two tests were...... complementary in the diagnosis of infection with S. pneumoniae. The third group of patients with probable pneumococcal infection were identified as 26% and 20% of the remaining 137 patients with unknown or known non-pneumococcal etiology, respectively, who received recent antibiotic treatment within 2-4 weeks...... of diagnostic sampling. By comparison, 0% (p < 0.01) with documented pneumococcal infection received antibiotic treatment in weeks 2-4 prior to microbiological sampling. As such a further eight patients should be expected to have infection with S. pneumoniae but would test negative in both culture...

  6. Serotype Distribution, Antibiotic Resistance and Clonality of Streptococcus pneumoniae Isolated from Immunocompromised Patients in Tunisia

    Raddaoui, Anis; Simões, Alexandra S.; Baaboura, Rekaya; Félix, Sofia; Achour, Wafa; Ben Othman, Tarek; Béjaoui, Mohamed; Sá-Leão, Raquel; Ben Hassen, Assia

    2015-01-01

    Background Pneumococcal disease, a major cause of morbidity and mortality globally, has higher incidence among young children, the elderly and the immunocompromised of all ages. In Tunisia, pneumococcal conjugate vaccines (PCVs) are not included in the national immunization program. Also, few studies have described the epidemiology of S. pneumoniae in this country and, in particular, no molecular typing studies have been performed. The aim of this study was to evaluate serotype distribution, ...

  7. KLF4-dependent IL-10 expression in lung epithelial cells by Streptococcus pneumoniae

    Steinicke, Robert

    2011-01-01

    Although the release of potent pro-inflammatory mediators is central for mounting an efficient host response in infections, excessive inflammation may lead to deleterious tissue damage. This is highlighted in severe pneumococcal pneumonia, in which the delicate balance among a robust inflammatory response to kill pneumococci and loss of organ function determines outcome of disease. Therefore we assessed the regulation of the potent anti-inflammatory cytokine IL-10 in pneumococci infection. S....

  8. Genome-wide identification of Streptococcus pneumoniae genes essential for bacterial replication during experimental meningitis

    Molzen, T E; Burghout, P; Bootsma, H J;

    2010-01-01

    of invasive pneumococcal disease is required in order to enable the development of new or adjunctive treatments and/or pneumococcal vaccines that are efficient across serotypes. We applied genomic array footprinting (GAF) in the search for S. pneumoniae genes that are essential during experimental...... relevant as targets for future therapy and prevention of pneumococcal meningitis, since their mutants were attenuated in both models of infection as well as in competitive growth in human cerebrospinal fluid in vitro....

  9. Phenotypic and Genotypic Characterization of Streptococcus pneumoniae Strains Colonizing Children Attending Day-Care Centers in Norway▿

    Vestrheim, Didrik F.; Høiby, E. Arne; Aaberge, Ingeborg S.; Caugant, Dominique A.

    2008-01-01

    A cross-sectional study of nasopharyngeal colonization with Streptococcus pneumoniae was performed among 573 children attending 29 day-care centers (DCCs) in Norway prior to the start of mass vaccination with the heptavalent pneumococcal conjugate vaccine (PCV-7). A sensitive sampling method was employed, including transport in an enrichment broth and serotyping of pneumococci directly from the broth, in addition to traditional single-colony isolation from blood agar plates. The prevalence of carriage was high, peaking at 88.7% in 2-year-olds. More than one serotype was isolated from 12.7% of the carriers. Of 509 isolates obtained, 227 (44.6%) belonged to the PCV-7 serotypes. Penicillin nonsusceptibility was rare (1.8% of the isolates). Nonsusceptibility to erythromycin (5.9%), clindamycin (2.0%), and tetracycline (5.5%) was associated with PCV-7 serotypes (P < 0.001). Multilocus sequence typing was performed on the whole strain collection, revealing 102 sequence types (STs), of which 31 (30.4%) were novel. Eleven isolates (2.2%) belonged to the England14-9 clone, and 19 isolates (3.7%) belonged to, or were single-locus variants of, the Portugal19F-21 clone. The pneumococcal populations within the DCCs were composed of a majority of isolates with STs shared between the DCCs and a minority of isolates with STs unique for each DCC. The highest numbers of different STs, including novel STs, were found within the most frequent serotypes. Our study indicates that carriage of S. pneumoniae is highly prevalent among children in Norwegian DCCs, with a genetically diverse pneumococcal population consisting of unique microepidemic DCC populations. PMID:18524970

  10. Phenotypic and genotypic characterization of Streptococcus pneumoniae strains colonizing children attending day-care centers in Norway.

    Vestrheim, Didrik F; Høiby, E Arne; Aaberge, Ingeborg S; Caugant, Dominique A

    2008-08-01

    A cross-sectional study of nasopharyngeal colonization with Streptococcus pneumoniae was performed among 573 children attending 29 day-care centers (DCCs) in Norway prior to the start of mass vaccination with the heptavalent pneumococcal conjugate vaccine (PCV-7). A sensitive sampling method was employed, including transport in an enrichment broth and serotyping of pneumococci directly from the broth, in addition to traditional single-colony isolation from blood agar plates. The prevalence of carriage was high, peaking at 88.7% in 2-year-olds. More than one serotype was isolated from 12.7% of the carriers. Of 509 isolates obtained, 227 (44.6%) belonged to the PCV-7 serotypes. Penicillin nonsusceptibility was rare (1.8% of the isolates). Nonsusceptibility to erythromycin (5.9%), clindamycin (2.0%), and tetracycline (5.5%) was associated with PCV-7 serotypes (P < 0.001). Multilocus sequence typing was performed on the whole strain collection, revealing 102 sequence types (STs), of which 31 (30.4%) were novel. Eleven isolates (2.2%) belonged to the England(14)-9 clone, and 19 isolates (3.7%) belonged to, or were single-locus variants of, the Portugal(19F)-21 clone. The pneumococcal populations within the DCCs were composed of a majority of isolates with STs shared between the DCCs and a minority of isolates with STs unique for each DCC. The highest numbers of different STs, including novel STs, were found within the most frequent serotypes. Our study indicates that carriage of S. pneumoniae is highly prevalent among children in Norwegian DCCs, with a genetically diverse pneumococcal population consisting of unique microepidemic DCC populations. PMID:18524970

  11. Identification of a human immunodominant B-cell epitope within the immunoglobulin A1 protease of Streptococcus pneumoniae

    De Paolis, Francesca; Beghetto, Elisa; Spadoni, Andrea; Montagnani, Francesca; Felici, Franco; Oggioni, Marco R; Gargano, Nicola

    2007-01-01

    Background The IgA1 protease of Streptococcus pneumoniae is a proteolytic enzyme that specifically cleaves the hinge regions of human IgA1, which dominates most mucosal surfaces and is the major IgA isotype in serum. This protease is expressed in all of the known pneumococcal strains and plays a major role in pathogen's resistance to the host immune response. The present work was focused at identifying the immunodominant regions of pneumococcal IgA1 protease recognized by the human antibody response. Results An antigenic sequence corresponding to amino acids 420–457 (epiA) of the iga gene product was identified by screening a pneumococcal phage display library with patients' sera. The epiA peptide is conserved in all pneumococci and in two out of three S. mitis strains, while it is not present in other oral streptococci so far sequenced. This epitope was specifically recognized by antibodies present in sera from 90% of healthy adults, thus representing an important target of the humoral response to S. pneumoniae and S. mitis infection. Moreover, sera from 68% of children less than 4 years old reacted with the epiA peptide, indicating that the human immune response against streptococcal antigens occurs during childhood. Conclusion The broad and specific recognition of the epiA polypeptide by human sera demonstrate that the pneumococcal IgA1 protease contains an immunodominant B-cell epitope. The use of phage display libraries to identify microbe or disease-specific antigens recognized by human sera is a valuable approach to epitope discovery. PMID:18088426

  12. Identification of a human immunodominant B-cell epitope within the immunoglobulin A1 protease of Streptococcus pneumoniae

    Felici Franco

    2007-12-01

    Full Text Available Abstract Background The IgA1 protease of Streptococcus pneumoniae is a proteolytic enzyme that specifically cleaves the hinge regions of human IgA1, which dominates most mucosal surfaces and is the major IgA isotype in serum. This protease is expressed in all of the known pneumococcal strains and plays a major role in pathogen's resistance to the host immune response. The present work was focused at identifying the immunodominant regions of pneumococcal IgA1 protease recognized by the human antibody response. Results An antigenic sequence corresponding to amino acids 420–457 (epiA of the iga gene product was identified by screening a pneumococcal phage display library with patients' sera. The epiA peptide is conserved in all pneumococci and in two out of three S. mitis strains, while it is not present in other oral streptococci so far sequenced. This epitope was specifically recognized by antibodies present in sera from 90% of healthy adults, thus representing an important target of the humoral response to S. pneumoniae and S. mitis infection. Moreover, sera from 68% of children less than 4 years old reacted with the epiA peptide, indicating that the human immune response against streptococcal antigens occurs during childhood. Conclusion The broad and specific recognition of the epiA polypeptide by human sera demonstrate that the pneumococcal IgA1 protease contains an immunodominant B-cell epitope. The use of phage display libraries to identify microbe or disease-specific antigens recognized by human sera is a valuable approach to epitope discovery.

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

    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.

  14. Serum level of YKL-40 is elevated in patients with Streptococcus pneumoniae bacteremia and is associated with the outcome of the disease

    Kronborg, Gitte; Ostergaard, Christian; Nielsen, Henrik; Obel, Niels; Pedersen, Svend S; Price, Paul A; Johansen, Julia S; Weis, Nina

    2002-01-01

    YKL40 is secreted by activated macrophages and neutrophils. Elevated serum concentrations of YKL40 are found in patients with diseases characterized by inflammation or ongoing fibrosis. The aim of this study was to evaluate serum YKL-40 levels in patients with Streptococcus pneumoniae bacteremia...... and to correlate these levels with clinical findings and outcomes. YKL40 was determined by ELISA and 89 patients were included in the study. Serum YKL-40 levels were significantly higher in patients with S. pneumoniae bacteremia (median 342 microg/l; range 20-20,400 microg/l) than in age...... was an independent prognostic factor of survival in logistic multivariate regression analysis (p = 0.002). In conclusion, high serum levels of YKL40 indicated a poorer prognosis for patients with S. pneumoniae bacteremia....

  15. In vitro activity of fluoroquinolones (gatifloxacin, levofloxacin and trovafloxacin and seven other antibiotics against Streptococcus pneumoniae

    Nicodemo A.C.

    2001-01-01

    Full Text Available In recent years, the level of resistance of S. pneumoniae to beta-lactam and/or macrolides has increased around the world including some countries in South America. Because of this resistance, it is necessary to test the therapeutic alternatives for treating this pathogen, including the newer quinolones. This study was carried out in order to compare the in vitro activity of fluoroquinolones gatifloxacin, levofloxacin and trovafloxacin, to penicillin G, amoxicillin, amoxicillin-clavulanate, cufuroxime sodium, ceftriaxone, azithromycin and clarithromycin, against 300 strains of S. pneumoniae. Of the 300 samples tested, 18.6% were not susceptible to penicillin (56 strains and 7% (21 strains were resistant to the second generation cephalosporin. Among the macrolides, resistance ranged from 6.7% for clarithromycin to 29.6% for azithromycin. Susceptibility to the newer quinolones was 100% including the 56 strains not susceptible to penicillin. Among the 10 antibiotics evaluated, the fluoroquinolones gatifloxacin, levofloxacin, and trovafloxacin displayed high levels of in vitro activity against S. pneumoniae.

  16. Colonização e resistência antimicrobiana de Streptococcus pneumoniae isolado em nasofaringe de crianças com rinofaringite aguda Nasopharyngeal colonization and antimicrobial resistance of Streptococcus pneumoniae isolated in children with acute rinofaringitis

    Lêda Lúcia M. Ferreira

    2001-06-01

    ças com infecção respiratória alta podem ser usados na vigilância da resistência antimicrobiana numa determinada comunidade.OBJECTIVE: to determine the prevalence and risk factors for nasopharyngeal colonization by, and to evaluate antimicrobial susceptibility of Streptococcus pneumoniae strains in children with acute rhinopharyngitis. METHODS: we collected nasopharyngeal swab specimens from 400 children aged 3 months to 5 years and with clinical status of acute rhinopharyngitis from June 16, 1997 to May 20, 1998 at the outpatient clinics of two hospitals in the city of São Paulo. Nasopharyngeal specimens were collected pernasally using a calcium alginate swab and plated immediately after collection onto trypticose soy agar with 5% sheep blood and garamicin 5 mcg/ml. Penicillin susceptibility was determined by oxacillin 1 mcg disk screening test and the minimal inhibitory concentration by the E-test. RESULTS: Pneumococci were recovered from 139 children, indicating a colonization prevalence of 35%. The risk factors analyzed indicated that the colonization was more prevalent in children attending day-care centers, children with siblings younger than 5 years, and children with recent use of antimicrobial agents. The prevalence of penicillin non-susceptible strains was of 16 % (20 strains. All strains were intermediately resistant (0.1mcg/ ml < MIC < 1.0 mcg/ ml. Out of the penicillin intermediately resistant strains, 7 (37% showed intermediate resistance to cotrimoxazol and 2 (11% full resistance to trimethoprim-sulfamethoxazole. No strains were resistant to ceftriaxone, amoxicillin, clarithromicin, or chloramphenicol. CONCLUSIONS: our findings indicate that the prevalence of nasopharyngeal colonization by Streptococcus pneumoniae in children with upper respiratory infections was of 34.8%. Children attending day-care centers and children with younger siblings showed higher levels of colonization The results of prevalence of bacterial resistance were similar to those

  17. Streptococcus pneumoniae and Haemophilus influenzae as etiological agents of conjunctivitis outbreaks in the region of Ribeirão Preto, SP, Brazil

    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.

  18. Streptococcus pneumoniae PstS production is phosphate responsive and enhanced during growth in the murine peritoneal cavity

    Orihuela, C. J.; Mills, J.; Robb, C. W.; Wilson, C. J.; Watson, D. A.; Niesel, D. W.

    2001-01-01

    Differential display-PCR (DDPCR) was used to identify a Streptococcus pneumoniae gene with enhanced transcription during growth in the murine peritoneal cavity. Northern dot blot analysis and comparative densitometry confirmed a 1.8-fold increase in expression of the encoded sequence following murine peritoneal culture (MPC) versus laboratory culture or control culture (CC). Sequencing and basic local alignment search tool analysis identified the DDPCR fragment as pstS, the phosphate-binding protein of a high-affinity phosphate uptake system. PCR amplification of the complete pstS gene followed by restriction analysis and sequencing suggests a high level of conservation between strains and serotypes. Quantitative immunodot blotting using antiserum to recombinant PstS (rPstS) demonstrated an approximately twofold increase in PstS production during MPC from that during CCs, a finding consistent with the low levels of phosphate observed in the peritoneum. Moreover, immunodot blot and Northern analysis demonstrated phosphate-dependent production of PstS in six of seven strains examined. These results identify pstS expression as responsive to the MPC environment and extracellular phosphate concentrations. Presently, it remains unclear if phosphate concentrations in vivo contribute to the regulation of pstS. Finally, polyclonal antiserum to rPstS did not inhibit growth of the pneumococcus in vitro, suggesting that antibodies do not block phosphate uptake; moreover, vaccination of mice with rPstS did not protect against intraperitoneal challenge as assessed by the 50% lethal dose.

  19. 99mTcN-gatifloxacin dithiocarbamate complex. A novel multi-drug-resistance Streptococcus pneumoniae (MRSP) infaction radiotracer

    Gatifloxacin (GTN) was derivatized to its dithiocarbamate derivative and its radiolabeling with technetium-99m (99mTc) using the [99mTc≡N]2+ core was investigated. The appropriateness of the 99mTcN - gatifloxacin dithiocarbamate (99mTcN - GTND) complex as a potential multi-drug-resistance Streptococcus pneumoniae (MRSP) infection radiotracer was evaluated in terms of stability in saline, serum, in vitro binding with MRSP and biodistribution in artificially MRSP infected Male Wistar Rats (MWR). In saline the 99mTcN - GTND complex showed more than 90% labeling yield up to 4 h with a maximum yield of 98.25 ± 0.20%, after reconstitution. In serum the 99mTcN - GTND complex showed stability up to 16 h of incubation with the appearance of insignificant 15.95% undesirable side products. The 99mTcN - GTND complex demonstrated saturated in vitro binding with MRSP with a maximum value of 75.50 ± 1.00% (at 90 min). In MWR model of group A, almost six times higher uptake of the labeled GTND was monitored in the muscle of MWR infected with live MRSP as compared to the inflamed and normal muscles. Based on the higher labeling yield in saline, in vitro stability in serum, saturated in vitro binding with live MRSP and promising biodistribution in MWR model we recommend 99mTcN - gatifloxacin dithiocarbamate complex as a potential MRSP infection radiotracer. (author)

  20. Structure of MurF from Streptococcus pneumoniae co-crystallized with a small molecule inhibitor exhibits interdomain closure

    Longenecker, Kenton L.; Stamper, Geoffrey F.; Hajduk, Philip J.; Fry, Elizabeth H.; Jakob, Clarissa G.; Harlan, John E.; Edalji, Rohinton; Bartley, Diane M.; Walter, Karl A.; Solomon, Larry R.; Holzman, Thomas F.; Gu, Yu Gui; Lerner, Claude G.; Beutel, Bruce A.; Stoll, Vincent S. (Abbott)

    2010-07-19

    In a broad genomics analysis to find novel protein targets for antibiotic discovery, MurF was identified as an essential gene product for Streptococcus pneumonia that catalyzes a critical reaction in the biosynthesis of the peptidoglycan in the formation of the cell wall. Lacking close relatives in mammalian biology, MurF presents attractive characteristics as a potential drug target. Initial screening of the Abbott small-molecule compound collection identified several compounds for further validation as pharmaceutical leads. Here we report the integrated efforts of NMR and X-ray crystallography, which reveal the multidomain structure of a MurF-inhibitor complex in a compact conformation that differs dramatically from related structures. The lead molecule is bound in the substrate-binding region and induces domain closure, suggestive of the domain arrangement for the as yet unobserved transition state conformation for MurF enzymes. The results form a basis for directed optimization of the compound lead by structure-based design to explore the suitability of MurF as a pharmaceutical target.

  1. Breakage-reunion domain of Streptococcus pneumoniae topoisomerase IV: crystal structure of a gram-positive quinolone target.

    Ivan Laponogov

    Full Text Available The 2.7 A crystal structure of the 55-kDa N-terminal breakage-reunion domain of topoisomerase (topo IV subunit A (ParC from Streptococcus pneumoniae, the first for the quinolone targets from a gram-positive bacterium, has been solved and reveals a 'closed' dimer similar in fold to Escherichia coli DNA gyrase subunit A (GyrA, but distinct from the 'open' gate structure of Escherichia coli ParC. Unlike GyrA whose DNA binding groove is largely positively charged, the DNA binding site of ParC exhibits a distinct pattern of alternating positively and negatively charged regions coincident with the predicted positions of the grooves and phosphate backbone of DNA. Based on the ParC structure, a new induced-fit model for sequence-specific recognition of the gate (G segment by ParC has been proposed. These features may account for the unique DNA recognition and quinolone targeting properties of pneumococcal type II topoisomerases compared to their gram-negative counterparts.

  2. Nanogel-based PspA intranasal vaccine prevents invasive disease and nasal colonization by Streptococcus pneumoniae.

    Kong, Il Gyu; Sato, Ayuko; Yuki, Yoshikazu; Nochi, Tomonori; Takahashi, Haruko; Sawada, Shinichi; Mejima, Mio; Kurokawa, Shiho; Okada, Kazunari; Sato, Shintaro; Briles, David E; Kunisawa, Jun; Inoue, Yusuke; Yamamoto, Masafumi; Akiyoshi, Kazunari; Kiyono, Hiroshi

    2013-05-01

    To establish a safer and more effective vaccine against pneumococcal respiratory infections, current knowledge regarding the antigens common among pneumococcal strains and improvements to the system for delivering these antigens across the mucosal barrier must be integrated. We developed a pneumococcal vaccine that combines the advantages of pneumococcal surface protein A (PspA) with a nontoxic intranasal vaccine delivery system based on a nanometer-sized hydrogel (nanogel) consisting of a cationic cholesteryl group-bearing pullulan (cCHP). The efficacy of the nanogel-based PspA nasal vaccine (cCHP-PspA) was tested in murine pneumococcal airway infection models. Intranasal vaccination with cCHP-PspA provided protective immunity against lethal challenge with Streptococcus pneumoniae Xen10, reduced colonization and invasion by bacteria in the upper and lower respiratory tracts, and induced systemic and nasal mucosal Th17 responses, high levels of PspA-specific serum immunoglobulin G (IgG), and nasal and bronchial IgA antibody responses. Moreover, there was no sign of PspA delivery by nanogel to either the olfactory bulbs or the central nervous system after intranasal administration. These results demonstrate the effectiveness and safety of the nanogel-based PspA nasal vaccine system as a universal mucosal vaccine against pneumococcal respiratory infection. PMID:23460513

  3. Alta prevalência de crianças portadoras de Streptococcus pneumoniae resistentes à penicilina em creches públicas High prevalence of children colonized with penicillin-resistant Streptococcus pneumoniae in public day-care centers

    Patrícia A. G. Velasquez

    2009-12-01

    Full Text Available OBJETIVOS: Investigar a prevalência de Streptococcus pneumoniae (pneumococos na nasofaringe de crianças sadias atendidas em creches municipais da cidade de Umuarama (PR. Avaliar a susceptibilidade aos antimicrobianos dos pneumococos isolados. MÉTODOS: Secreção da nasofaringe de 212 crianças foi coletada no período de abril a outubro de 2008. Após semeadura dos espécimes em ágar sangue e incubação a 37 °C por 24-48 horas, as colônias suspeitas de pertencerem a S. pneumoniae foram identificadas pela α-hemólise, sensibilidade à optoquina e bile solubilidade. A susceptibilidade à penicilina foi investigada pelos testes de disco-difusão e de diluição. A susceptibilidade aos demais antimicrobianos indicados no tratamento das infecções pneumocócicas foi realizada por disco-difusão RESULTADOS: A prevalência de pneumococos na nasofaringe foi de 43,4% (92/212, sendo maior em crianças com idade entre 2 e 5 anos (p = 0,0005. Não houve diferença significativa entre os sexos. Resistência intermediária e resistência plena à penicilina foram encontradas respectivamente em 34,8 (32/92 e 22,8% (21/92 dos isolados. Sessenta e sete amostras (72,8% foram resistentes ao sulfametoxazol-trimetoprim, oito (8,7% à eritromicina e seis (6,5% à tetraciclina. Uma amostra apresentou resistência à clindamicina (1,1%, e outra ao cloranfenicol (1,1%. Todas as amostras foram sensíveis a levofloxacina, ofloxacina, rifampicina, telitromicina, linezolide e vancomicina. Nove amostras foram consideradas multirresistentes, por apresentarem resistência a três ou mais classes de antimicrobianos. CONCLUSÕES: O presente estudo registrou uma alta prevalência de crianças portadoras sadias de amostras de S. pneumoniae resistentes à penicilina que podem constituir importantes reservatórios desse patógeno na comunidade.OBJECTIVES: To investigate the prevalence of Streptococcus pneumoniae (pneumococci in the nasopharynx of healthy children enrolled

  4. In vitro activity of Tedizolid phosphate against multidrug-resistant Streptococcus pneumoniae isolates from Asian countries.

    Baek, Jin Yang; Kang, Cheol-In; Kim, So Hyun; Ko, Kwan Soo; Chung, Doo Ryeon; Peck, Kyong Ran; Hsueh, Po-Ren; Thamlikitkul, Visanu; So, Thomas Man-Kit; Lee, Nam Yong; Song, Jae-Hoon

    2016-06-01

    Tedizolid phosphate is a second-generation oxazolidinone prodrug that is potential activity against a wide range of Gram-positive pathogens, including methicillin-resistant Staphylococcus aureus, penicillin-resistant streptococci, and vancomycin-resistant enterococci. The in vitro activity of tedizolid and other comparator agents against multidrug-resistant (MDR) pneumococci from various Asian countries were evaluated. Of the S. pneumoniae clinical pneumonia isolates collected during 2008 and 2009 from 8 Asian countries (Korea, Taiwan, Thailand, Hong Kong, Vietnam, Malaysia, Philippines, and Sri Lanka), 104 isolates of MDR pneumococci were included in this study. Antimicrobial susceptibility testing for 18 antimicrobial agents was performed by broth microdilution method. Tedizolid was highly active against pneumococci. All isolates tested were inhibited at a tedizolid minimum inhibitory concentration (MIC) value of ≤0.25μg/ml (ranged from ≤0.03μg/ml to 0.25μg/ml). The MIC50 and MIC90 of tedizolid against MDR pneumococci were both 0.12μg/ml, while MIC50 and MIC90 of linezolid were 0.5μg/ml and 1μg/ml, respectively. In addition, tedizolid maintained the activity against S. pneumoniae regardless of the extensively drug-resistant (XDR) phenotype of the isolates. The activity of tedizolid was excellent against all types of MDR pneumococci, exhibiting and maintaining at least 4-fold-greater potency compared to linezolid, regardless of resistance phenotypes to other commonly utilized agents. Tedizolid has the potential to be an agent to treat infections caused by MDR pneumococci in the Asia. PMID:27083121

  5. Multiplex quantitative PCR for detection of lower respiratory tract infection and meningitis caused by Streptococcus pneumoniae, Haemophilus influenzae and Neisseria meningitidis

    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

  6. Pneumonia in Pregnancy

    Maurizio Maccato

    1995-01-01

    Pneumonia complicating pregnancy requires a prompt diagnosis and the institution of adequate supportive and antimicrobial therapy. In a patient with a classic presentation of pneumonia, the most likely pathogens are Streptococcus pneumoniae and Haemophilus influenzae. In a patient with an atypical presentation of pneumonia, Mycoplasma pneumoniae and Chlamydia pneumoniae are frequently encountered. In a patient suffering from acquired immunodeficiency syndrome (AIDS), Pneumocystis carinii is t...

  7. Establishment and Application of AFLP Method for Rapid Diagnosis of Streptococcus Pneumoniae%肺炎链球菌的扩增片段长度多态性快速检测方法的建立和应用

    杨锦红; 刘洋; 王新林; 李方去; 陶洪群; 李向阳

    2012-01-01

    目的 建立扩增片段长度多态性(AFLP)快速检测肺炎链球菌.方法 针对肺炎链球菌的管家基因16SrRNA基因设计特异引物,优化反应条件,建立肺炎链球菌的AFLP检测方法.通过对20株肺炎链球菌和15株非肺炎链球菌进行检测,检验该方法的灵敏度、特异度和实用性.结果 活菌计数方法表明建立的AFLP方法检测肺炎链球菌的灵敏度为1.5×103CFU/ml.检测20株肺炎链球菌均为阳性,15株非肺炎链球菌则全部阴性,特异度为100%.结论 建立的AFLP方法检测肺炎链球菌灵敏度、特异度高,可用于肺炎链球菌的检测和流行病学调查.%Objective To establish a amplified fragment length polymorphism (AFLP) method for rapid diagnosis of Streptococcus pneumonia. Methods Based on the 16SrRNA nucleic sequence of streptococcus pneumonia,a pair of primers was designed for AFLP. The reaction conditions were optimized, and the specificity, sensitivity, and practicability of AFLP were tested using 20 streptococcus pneumonia strains and IS non - streptococcus pneumonia strainss in blood. Results The results of streptococcus pneumonia count showed that AFLP was capable of detecting streptococcus pneumonia at a level as low as 1.5 ×103CFU/ml. All the 20 strains of streptococcus pneumonia yielded positive results in AFLP,and the 15 strains of other bacteria all showed negative results, with a detection specificity of 100% . Conclusion The established AFLP method has high specificity and sensitivity for detecting streptococcus pneumonia and is applicable in field monitoring and epidemiological study of streptococcus pneumonia.

  8. Status of research and development of pediatric vaccines for Streptococcus pneumoniae.

    Alderson, Mark R

    2016-06-01

    Pneumococcal disease is a major cause of morbidity and mortality in young children, particularly in the developing world. Vaccines are a critical strategy for protecting children from pneumococcal disease and licensed pneumococcal conjugate vaccines (PCVs) are having a significant impact on invasive pneumococcal disease and pneumococcal pneumonia throughout the world. Currently available PCVs do not, however, cover all pneumococcal serotypes and are complicated and relatively expensive to manufacture. While new PCV development is focused on either higher valency or more inherent affordability for developing countries, new vaccines are needed that offer serotype-independent protection. Vaccines containing proteins that are common to all pneumococcal serotypes could provide broad protection to children worldwide. Protein subunit and whole cell vaccines have advanced into Phase 1 and 2 clinical trials but face considerable challenges before they can become licensed and widely distributed. PMID:27083428

  9. Endocarditis in Greenland with special reference to endocarditis caused by Streptococcus pneumoniae

    Madsen, Rasmus Gaarde; Ladefoged, Karin; Kjaergaard, Jens Jørgen;

    2009-01-01

    in studies on Caucasian populations, where pneumococcal infection was seen in 1-3% of endocarditis cases. The overall mortality rate was 12%. Pneumococcal endocarditis (PE) had the clinical characteristics of fulminant disease with frequent heart failure, complications and need for surgery. Among......OBJECTIVES: The aim of this retrospective study was to determine the incidence and outcome of infectious endocarditis in Greenland with an emphasis on pneumococcal endocarditis. STUDY DESIGN: Retrospective, non-interventional study. METHODS: Review of files and medical history of all patients with...... infectious endocarditis from the Patient Registry in Greenland in the 11-year period 1995-2005. RESULTS: There were 25 cases of endocarditis, giving an incidence rate of 4.0/100,000 per year. Twenty-four percent of these cases were caused by Streptoccous pneumonia, which is significantly more frequent than...

  10. Progress of streptococcus pneumoniae infection and immunity%肺炎链球菌的感染与免疫研究进展

    赵冰(综述); 潘家华(审校)

    2014-01-01

    肺炎链球菌是儿童侵袭性细菌感染的主要病原菌,可以引起肺炎、脑膜炎和脓毒症等危及生命的疾病。作为上呼吸道常见定植菌,了解其感染过程中机体的免疫反应对于疾病的治疗至关重要。该文综述肺炎链球菌抗原结构、肺炎链球菌结合疫苗及主要毒力因子,并重点描述肺炎链球菌感染后机体的免疫反应,包括固有免疫、巨噬细胞、中性粒细胞和T细胞的作用。%Streptococcus pneumoniae( SP) is the leading pathogenic bacteria of invasive bacterial infec-tions in children. It can cause some life-threatening diseases such as pneumonia,meningitis and sepsis. Strepto-coccus pneumoniae frequently colonizes the upper respiratory tract. It is essential to know the host immune re-sponse during the infection. This paper reviews antigenic structure of SP,pneumococcal conjugate vaccine and pneumococcal virulence factors,especially focuses on the immune response including the effect of innate immu-nity,macrophage,neutrophil and T-cell.

  11. Low Concentrations of Nitric Oxide Modulate Streptococcus pneumoniae Biofilm Metabolism and Antibiotic Tolerance.

    Allan, Raymond N; Morgan, Samantha; Brito-Mutunayagam, Sanjita; Skipp, Paul; Feelisch, Martin; Hayes, Stephen M; Hellier, William; Clarke, Stuart C; Stoodley, Paul; Burgess, Andrea; Ismail-Koch, Hasnaa; Salib, Rami J; Webb, Jeremy S; Faust, Saul N; Hall-Stoodley, Luanne

    2016-04-01

    Streptococcus pneumoniaeis one of the key pathogens responsible for otitis media (OM), the most common infection in children and the largest cause of childhood antibiotic prescription. Novel therapeutic strategies that reduce the overall antibiotic consumption due to OM are required because, although widespread pneumococcal conjugate immunization has controlled invasive pneumococcal disease, overall OM incidence has not decreased. Biofilm formation represents an important phenotype contributing to the antibiotic tolerance and persistence ofS. pneumoniaein chronic or recurrent OM. We investigated the treatment of pneumococcal biofilms with nitric oxide (NO), an endogenous signaling molecule and therapeutic agent that has been demonstrated to trigger biofilm dispersal in other bacterial species. We hypothesized that addition of low concentrations of NO to pneumococcal biofilms would improve antibiotic efficacy and that higher concentrations exert direct antibacterial effects. Unlike in many other bacterial species, low concentrations of NO did not result inS. pneumoniaebiofilm dispersal. Instead, treatment of bothin vitrobiofilms andex vivoadenoid tissue samples (a reservoir forS. pneumoniaebiofilms) with low concentrations of NO enhanced pneumococcal killing when combined with amoxicillin-clavulanic acid, an antibiotic commonly used to treat chronic OM. Quantitative proteomic analysis using iTRAQ (isobaric tag for relative and absolute quantitation) identified 13 proteins that were differentially expressed following low-concentration NO treatment, 85% of which function in metabolism or translation. Treatment with low-concentration NO, therefore, appears to modulate pneumococcal metabolism and may represent a novel therapeutic approach to reduce antibiotic tolerance in pneumococcal biofilms. PMID:26856845

  12. Surface Proteins and Pneumolysin of Encapsulated and Nonencapsulated Streptococcus pneumoniae Mediate Virulence in a Chinchilla Model of Otitis Media

    Keller, Lance E.; Bradshaw, Jessica L.; Pipkins, Haley; McDaniel, Larry S.

    2016-01-01

    Streptococcus pneumoniae infections result in a range of human diseases and are responsible for almost one million deaths annually. Pneumococcal disease is mediated in part through surface structures and an anti-phagocytic capsule. Recent studies have shown that nonencapsulated S. pneumoniae (NESp) make up a significant portion of the pneumococcal population and are able to cause disease. NESp lack some common surface proteins expressed by encapsulated pneumococci, but express surface proteins unique to NESp. A chinchilla model of otitis media (OM) was used to determine the effect various pneumococcal mutations have on pathogenesis in both NESp and encapsulated pneumococci. Epithelial cell adhesion and invasion assays were used to examine the effects in relation to deletion of intrinsic genes or expression of novel genes. A mouse model of colonization was also utilized for comparison of various pneumococcal mutants. It was determined that pneumococcal surface protein K (PspK) and pneumolysin (Ply) affect NESp middle ear pathogenesis, but only PspK affected epithelial cell adhesion. Experiments in an OM model were done with encapsulated strains testing the importance of native virulence factors and treatment of OM. First, a triple deletion of the common virulence factors PspA, PspC, and Ply, (ΔPAC), from an encapsulated background abolished virulence in an OM model while a PspC mutant had detectable, but reduced amounts of recoverable bacteria compared to wildtype. Next, treatment of OM was effective when starting antibiotic treatment within 24 h with resolution by 48 h post-treatment. Expression of NESp-specific virulence factor PspK in an encapsulated strain has not been previously studied, and we showed significantly increased adhesion and invasion of human epithelial cells by pneumococci. Murine colonization was not significantly increased when an encapsulated strain expressed PspK, but colonization was increased when a capsule mutant expressed PspK. The

  13. [Sensitivity surveillance of Streptococcus pneumoniae isolates for several antibacterial agents in Gifu and Aichi prefectures (2011-2012)].

    Funatsu, Tori; Mizunaga, Shingo; Fukuda, Yoshiko; Nomura, Nobuhiko; Hashido, Hikonori; Mitsuyama, Junichi; Hatano, Masakazu; Yamaoka, Kazukiyo; Watanabe, Kunitomo; Asano, Yuko; Suematsu, Hiroyuki; Sawamura, Haruki; Matsukawa, Yoko; Ohta, Hirotoshi; Yamagishi, Yuka; Mikamo, Hiroshige; Matsubara, Shigenori; Shibata, Naohiro

    2015-08-01

    We investigated the susceptibility to antibacterial agents, genotype of penicillin-binding protein (PBP) genes and macrolide resistant genes, and the serotypes against 270 strains of Streptococcus pneumoniae isolated from medical facilities in Gifu and Aichi prefectures between October 2011 and April 2012. These results were compared with those against S. pneumoniae isolated in 2008-2009 and 2010-2011. The number of gPSSP with 3 normal PBP genes, gPISP with 1 or 2 normal PBP genes and gPRSP with 3 abnormal genes isolated in 2011-2012 was 15 (5.6%), 162 (60.0%) and 93 (34.4%) strains, respectively. Compared with those isolated in 2008-2009 and 2010-2011, the numbers of gPRSP were decreasing. On the other hand, the isolates with no macrolide-resistant gene, only mefA, only ermB, and both mefA and ermB were 16 (5.9%), 75 (27.8%), 153 (56.7%) and 26 (9.6%). Compared with those isolated in 2008-2009 and 2010-2011, the numbers of isolates with ermB, which was usually associated with high-level resistance, were increasing. The prevalent pneumococcal serotypes in children were type 3 (14.4%), following by type 15 and 19F (9.3%). The coverages of 7-valent pneumococcal conjugate vaccine (PCV7) and 13-valent pneumococcal conjugate vaccine (PCV13) were calculated as 22.9% and 49.2%, respectively. The coverages of PCV7 and PCV13 in gPRSP isolated from children were 47.7% (21/44 strains) and 72.7% (32/44 strains). The MIC90 of each antibacterial agent was as follows; 0.125pg/mL for imipenem, panipenem and garenoxacin, 0.25 μg/mL for meropenem and doripenem, 0.5 μg/mL for cefditoren, moxifloxacin and tosufloxacin, 1 μg/mL for amoxicillin, clavulanic acid/amoxicillin, cefteram, cefcapene and ceftriaxone, 2 μg/mL for benzylpenicillin, ampicillin, sulbactam/ampicillin, piperacillin, tazobactam/piperacillin and levofloxacin, 4 μg/mL for cefdinir, flomoxef and pazufloxacin, 16 μg/mL for minocycline, > 64 μg/mL for clarithromycin and azithromycin, and these MIC90s were about the

  14. Surface Proteins and Pneumolysin of Encapsulated and Nonencapsulated Streptococcus pneumoniae Mediate Virulence in a Chinchilla Model of Otitis Media.

    Keller, Lance E; Bradshaw, Jessica L; Pipkins, Haley; McDaniel, Larry S

    2016-01-01

    Streptococcus pneumoniae infections result in a range of human diseases and are responsible for almost one million deaths annually. Pneumococcal disease is mediated in part through surface structures and an anti-phagocytic capsule. Recent studies have shown that nonencapsulated S. pneumoniae (NESp) make up a significant portion of the pneumococcal population and are able to cause disease. NESp lack some common surface proteins expressed by encapsulated pneumococci, but express surface proteins unique to NESp. A chinchilla model of otitis media (OM) was used to determine the effect various pneumococcal mutations have on pathogenesis in both NESp and encapsulated pneumococci. Epithelial cell adhesion and invasion assays were used to examine the effects in relation to deletion of intrinsic genes or expression of novel genes. A mouse model of colonization was also utilized for comparison of various pneumococcal mutants. It was determined that pneumococcal surface protein K (PspK) and pneumolysin (Ply) affect NESp middle ear pathogenesis, but only PspK affected epithelial cell adhesion. Experiments in an OM model were done with encapsulated strains testing the importance of native virulence factors and treatment of OM. First, a triple deletion of the common virulence factors PspA, PspC, and Ply, (ΔPAC), from an encapsulated background abolished virulence in an OM model while a PspC mutant had detectable, but reduced amounts of recoverable bacteria compared to wildtype. Next, treatment of OM was effective when starting antibiotic treatment within 24 h with resolution by 48 h post-treatment. Expression of NESp-specific virulence factor PspK in an encapsulated strain has not been previously studied, and we showed significantly increased adhesion and invasion of human epithelial cells by pneumococci. Murine colonization was not significantly increased when an encapsulated strain expressed PspK, but colonization was increased when a capsule mutant expressed PspK. The

  15. Identification of ComW as a new component in the regulation of genetic transformation in Streptococcus pneumoniae.

    Luo, Ping; Li, Haiying; Morrison, Donald A

    2004-10-01

    Regulation of competence for genetic transformation in Streptococcus pneumoniae depends on a quorum-sensing system, genes involved in DNA uptake and recombination and a link between these two gene sets. The alternative sigma factor ComX provides this link. ComE, the response regulator of the quorum-sensing system, is required for expression of ComX and other early genes. However, an unknown ComE-dependent regulator is also required for competence when comX is expressed under control of the raffinose-responsive promoter of the aga operon. The gene comW (SP0018) is required for a high level of competence and is regulated by the quorum-sensing system, but its function is unknown. To explore its role further, comW was cloned into the multicopy plasmid pMSP3535, under the control of a nisin-inducible promoter (P(N)), and transformed into pneumococcal strains containing a raffinose-inducible comX gene (P(R)::comX). Further introduction of a comE deletion blocked the endogenous CSP signal transduction pathway. In the resulting strain, competence was independent of CSP but depended on treatment with both nisin and raffinose, showing that coexpression of comW and comX complemented the comE deficiency. ComX protein accumulation and expression of a late competence gene in the above strain support the conclusion that ComW is a new positive factor involved in competence regulation. PMID:15458414

  16. Induction of prophages by fluoroquinolones in Streptococcus pneumoniae: implications for emergence of resistance in genetically-related clones.

    Elena López

    Full Text Available Antibiotic resistance in Streptococcus pneumoniae has increased worldwide by the spread of a few clones. Fluoroquinolone resistance occurs mainly by alteration of their intracellular targets, the type II DNA topoisomerases, which is acquired either by point mutation or by recombination. Increase in fluoroquinolone-resistance may depend on the balance between antibiotic consumption and the cost that resistance imposes to bacterial fitness. In addition, pneumococcal prophages could play an important role. Prophage induction by fluoroquinolones was confirmed in 4 clinical isolates by using Southern blot hybridization. Clinical isolates (105 fluoroquinolone-resistant and 160 fluoroquinolone-susceptible were tested for lysogeny by using a PCR assay and functional prophage carriage was studied by mitomycin C induction. Fluoroquinolone-resistant strains harbored fewer inducible prophages (17/43 than fluoroquinolone-susceptible strains (49/70 (P = 0.0018. In addition, isolates of clones associated with fluoroquinolone resistance [CC156 (3/25; CC63 (2/20, and CC81 (1/19], had lower frequency of functional prophages than isolates of clones with low incidence of fluoroquinolone resistance [CC30 (4/21, CC230 (5/20, CC62 (9/21, and CC180 (21/30]. Likewise, persistent strains from patients with chronic respiratory diseases subjected to fluoroquinolone treatment had a low frequency of inducible prophages (1/11. Development of ciprofloxacin resistance was tested with two isogenic strains, one lysogenic and the other non-lysogenic: emergence of resistance was only observed in the non-lysogenic strain. These results are compatible with the lysis of lysogenic isolates receiving fluoroquinolones before the development of resistance and explain the inverse relation between presence of inducible prophages and fluoroquinolone-resistance.

  17. Streptococcus pneumoniae Endohexosaminidase D, Structural and Mechanistic Insight into Substrate-Assisted Catalysis in Family 85 Glycoside Hydrolases

    Endo-?-d-glucosaminidases from family 85 of glycoside hydrolases (GH85 endohexosaminidases) act to cleave the glycosidic linkage between the two N-acetylglucosamine units that make up the chitobiose core of N-glycans. Endohexosaminidase D (Endo-D), produced by Streptococcus pneumoniae, is believed to contribute to the virulence of this organism by playing a role in the deglycosylation of IgG antibodies. Endohexosaminidases have received significant attention for this reason and, moreover, because they are powerful tools for chemoenzymatic synthesis of proteins having defined glycoforms. Here we describe mechanistic and structural studies of the catalytic domain (SpGH85) of Endo-D that provide compelling support for GH85 enzymes using a catalytic mechanism involving substrate-assisted catalysis. Furthermore, the structure of SpGH85 in complex with the mechanism-based competitive inhibitor NAG-thiazoline (Kd = 28 ?m) provides a coherent rationale for previous mutagenesis studies of Endo-D and other related GH85 enzymes. We also find GH85, GH56, and GH18 enzymes have a similar configuration of catalytic residues. Notably, GH85 enzymes have an asparagine in place of the aspartate residue found in these other families of glycosidases. We propose that this residue, as the imidic acid tautomer, acts analogously to the key catalytic aspartate of GH56 and GH18 enzymes. This topographically conserved arrangement of the asparagine residue and a conserved glutamic acid, coupled with previous kinetic studies, suggests these enzymes may use an unusual proton shuttle to coordinate effective general acid and base catalysis to aid cleavage of the glycosidic bond. These results collectively provide a blueprint that may be used to facilitate protein engineering of these enzymes to improve their function as biocatalysts for synthesizing glycoproteins having defined glycoforms and also may serve as a guide for generating inhibitors of GH85 enzymes.

  18. Streptococcus pneumoniae Endohexosaminidase D, Structural and Mechanistic Insight into Substrate-Assisted Catalysis in Family 85 Glycoside Hydrolases

    Abbott, D.; Macauley, M; Vocadlo, D; Boraston, A

    2009-01-01

    Endo-?-d-glucosaminidases from family 85 of glycoside hydrolases (GH85 endohexosaminidases) act to cleave the glycosidic linkage between the two N-acetylglucosamine units that make up the chitobiose core of N-glycans. Endohexosaminidase D (Endo-D), produced by Streptococcus pneumoniae, is believed to contribute to the virulence of this organism by playing a role in the deglycosylation of IgG antibodies. Endohexosaminidases have received significant attention for this reason and, moreover, because they are powerful tools for chemoenzymatic synthesis of proteins having defined glycoforms. Here we describe mechanistic and structural studies of the catalytic domain (SpGH85) of Endo-D that provide compelling support for GH85 enzymes using a catalytic mechanism involving substrate-assisted catalysis. Furthermore, the structure of SpGH85 in complex with the mechanism-based competitive inhibitor NAG-thiazoline (Kd = 28 ?m) provides a coherent rationale for previous mutagenesis studies of Endo-D and other related GH85 enzymes. We also find GH85, GH56, and GH18 enzymes have a similar configuration of catalytic residues. Notably, GH85 enzymes have an asparagine in place of the aspartate residue found in these other families of glycosidases. We propose that this residue, as the imidic acid tautomer, acts analogously to the key catalytic aspartate of GH56 and GH18 enzymes. This topographically conserved arrangement of the asparagine residue and a conserved glutamic acid, coupled with previous kinetic studies, suggests these enzymes may use an unusual proton shuttle to coordinate effective general acid and base catalysis to aid cleavage of the glycosidic bond. These results collectively provide a blueprint that may be used to facilitate protein engineering of these enzymes to improve their function as biocatalysts for synthesizing glycoproteins having defined glycoforms and also may serve as a guide for generating inhibitors of GH85 enzymes.

  19. Prevalence and clonal distribution of pcpA, psrP and Pilus-1 among pediatric isolates of Streptococcus pneumoniae.

    Laura Selva

    Full Text Available Streptococcus pneumoniae is the leading cause of vaccine-preventable deaths globally. The objective of this study was to determine the distribution and clonal type variability of three potential vaccine antigens: Pneumococcal serine-rich repeat protein (PsrP, Pilus-1, and Pneumococcal choline binding protein A (PcpA among pneumococcal isolates from children with invasive pneumococcal disease and healthy nasopharyngeal carriers. We studied by Real-Time PCR a total of 458 invasive pneumococcal isolates and 89 nasopharyngeal pneumococcal isolates among children (total = 547 strains collected in Barcelona, Spain, from January 2004 to July 2010. pcpA, psrP and pilus-1 were detected in 92.8%, 51.7% and 14.4% of invasive isolates and in 92.1%, 48.3% and 18% of carrier isolates, respectively. Within individual serotypes the prevalence of psrP and pilus-1 was highly dependent on the clonal type. pcpA was highly prevalent in all strains with the exception of those belonging to serotype 3 (33.3% in serotype 3 isolates vs. 95.1% in other serotypes; P<.001. psrP was significantly more frequent in those serotypes that are less apt to be detected in carriage than in disease; 58.7% vs. 39.1% P<.001. Antibiotic resistance was associated with the presence of pilus-1 and showed a negative correlation with psrP. These results indicate that PcpA, and subsequently Psrp and Pilus-1 together might be good candidates to be used in a next-generation of multivalent pneumococcal protein vaccine.

  20. Generation of genic diversity among Streptococcus pneumoniae strains via horizontal gene transfer during a chronic polyclonal pediatric infection.

    N Luisa Hiller

    Full Text Available Although there is tremendous interest in understanding the evolutionary roles of horizontal gene transfer (HGT processes that occur during chronic polyclonal infections, to date there have been few studies that directly address this topic. We have characterized multiple HGT events that most likely occurred during polyclonal infection among nasopharyngeal strains of Streptococcus pneumoniae recovered from a child suffering from chronic upper respiratory and middle-ear infections. Whole genome sequencing and comparative genomics were performed on six isolates collected during symptomatic episodes over a period of seven months. From these comparisons we determined that five of the isolates were genetically highly similar and likely represented a dominant lineage. We analyzed all genic and allelic differences among all six isolates and found that all differences tended to occur within contiguous genomic blocks, suggestive of strain evolution by homologous recombination. From these analyses we identified three strains (two of which were recovered on two different occasions that appear to have been derived sequentially, one from the next, each by multiple recombination events. We also identified a fourth strain that contains many of the genomic segments that differentiate the three highly related strains from one another, and have hypothesized that this fourth strain may have served as a donor multiple times in the evolution of the dominant strain line. The variations among the parent, daughter, and grand-daughter recombinant strains collectively cover greater than seven percent of the genome and are grouped into 23 chromosomal clusters. While capturing in vivo HGT, these data support the distributed genome hypothesis and suggest that a single competence event in pneumococci can result in the replacement of DNA at multiple non-adjacent loci.

  1. Current status of drug resistance ofStreptococcus pneumoniae isolated from children in China%儿童肺炎链球菌耐药现状

    葛玲丽; 韩志英

    2016-01-01

    Streptococcus pneumoniae is the most common bacterial pathogen of community- acquired infections in children, and antibiotics are the effective way of treatment for pneumococcal disease. Disease burden of pneumococcal infections has increased due to enhanced antibiotic resistance of Streptococcus pneumoniae, and it has a large regional differences. In recent years, surveillance for bacterial resistance ofStreptococcus pneumoniaeamong children has enhanced, and the related researches are increasing in China. This article provided an overview of the current status of drug resistance of Streptococcus pneumoniaeto β-lactam antibiotics, macrolide antibiotics and other antibiotics in children in China.%肺炎链球菌是儿童社区获得性感染最常见细菌病原,抗菌药物是治疗肺炎链球菌疾病的有效手段。由于抗生素的长期过度使用,肺炎链球菌对抗生素的耐药性逐渐增强,肺炎链球菌感染性疾病造成的负担逐渐加重,且不同地区差异较大。近年来国内对儿童肺炎链球菌抗生素耐药性的监测加强,文献报道增多,此文就国内儿童肺炎链球菌对β-内酰胺类、大环内酯类及其他种类抗生素的耐药状况进行综述。

  2. Phenotypes and genotypes of macrolide-resistant Streptococcus pneumoniae in Serbia

    Hadnađev Mirjana

    2014-01-01

    Full Text Available Although macrolides are widely used for treating pneumococcal infections, an increase in macrolide resistance might compromise their use. The objective of this study was to determine the prevalence of macrolide-resistant phenotypes and genotypes in macrolide-resistant S. pneumoniae isolates in Serbia. A total of 228 macrolide-resistant strains isolated during the period of 2009-2012, were analyzed. Macrolide resistance phenotypes were determined by a double disk diffusion test. The presence of macrolide resistance genes was detected by PCR. Antibiotics susceptibilities were tested using the VITEK2 system and E test. Among the examined isolates, the MLSB phenotype which is linked to the presence of the erm(B gene dominated (83.3%, while the mef(A gene which is associated with the M phenotype, was identified in 16.7% isolates. Over 40% of isolates expressed co-resistance to penicillin. A multiple-resistant pattern was found in 36.4% strains, more frequently in children. However, all strains were susceptible to telithromycin, vancomycin, linezolid, fluoroquinolones and rifampicin. [Projekat Ministarstva nauke Republike Srbije, br. 175039

  3. Serotype distribution of Streptococcus pneumoniae causing invasive disease in the Republic of Ireland.

    Vickers, I

    2011-05-01

    The 7-valent pneumococcal conjugate vaccine (PCV7) was included in the routine infant immunization schedule in Ireland in September 2008. We determined the serotype of 977 S. pneumoniae isolates causing invasive disease between 2000-2002 and 2007-2008, assessed for the presence of the recently described serotype 6C and determined the susceptibility of isolates during 2007-2008 to penicillin and cefotaxime. Serotype 14 was the most common serotype during both periods and 7·7% of isolates previously typed as serotype 6A were serotype 6C. During 2000-2002 and 2007-2008, PCV7 could potentially have prevented 85% and 74% of invasive pneumococcal disease in the target population (i.e. children aged <2 years), respectively. The level of penicillin non-susceptibility was 17% in 2007-2008. Ongoing surveillance of serotypes is required to determine the impact of PCV7 in the Irish population and to assess the potential of new vaccines with expanded valency.

  4. X-ray crystal structure of N-6 adenine deoxyribose nucleic acid methyltransferase from Streptococcus pneumoniae

    Tran, Phidung Hong

    X-ray diffraction by using resonant anomalous scattering has become a popular tool for solving crystal structures in the last ten years with the expanded availability of tunable synchrotron radiation for protein crystallography. Mercury atoms were used for phasing. The crystal structure of N-6 deoxyribose nucleic acid methyltransferase from Streptoccocus pneumoniae (DpnM) was solved by using the Multiple Anomalous Diffraction technique. The crystal structure reveals the formation of mercaptide between the mercury ion and the thiol group on the cysteine amino acid in a hydrophobic environment. The crystal structure contains the bound ligand, S- adenosyl-l-methionine on the surface of the concave opening. The direction of the β-strands on the beta sheets are identical to other solved methyltransferases. The highly conserved motifs, DPPY and the FxGxG, are found to be important in ligand binding and possibly in methyl group transfer. The structure has a concave cleft with an opening on the order of 30 Å that can accommodate a DNA duplex. By molecular modelling coupled to sequence alignment, two other highly conserved residues Arg21 and Gly19 are found to be important in catalysis.

  5. Distribuição e diversidade de elementos genéticos associados à virulência em Streptococcus pneumoniae

    Lopes, Joana Gomes Martins, 1989-

    2013-01-01

    Streptococcus pneumoniae é uma bactéria de Gram-positivo e um microrganismo comensal que coloniza assintomaticamente o aparelho respiratório superior (nasofaringe), contudo pode causar ocasionalmente infeção no hospedeiro com particular incidência nas crianças, idosos e doentes com condições crónicas debilitantes. Devido à sua grande variabilidade genética, resultante de altas taxas de transferência horizontal de genes, existe um desigual potencial patogénico entre estirpes. Entender como a v...

  6. Quinupristin-Dalfopristin Resistance in Streptococcus pneumoniae: Novel L22 Ribosomal Protein Mutation in Two Clinical Isolates from the SENTRY Antimicrobial Surveillance Program

    Ronald N Jones; Farrell, David J.; Morrissey, Ian

    2003-01-01

    Resistance to quinupristin-dalfopristin (Q/D) among gram-positive cocci has been very uncommon. Two clinical isolates among 8,837 (0.02%) Streptococcus pneumoniae isolates were discovered in 2001 to 2002 with Q/D MICs of 4 μg/ml. Each had a 5-amino-acid tandem duplication (RTAHI) in the L22 ribosomal protein gene (rplV) preventing synergistic ribosomal binding of the streptogramin combination. Similar gene duplication has been reported in Q/D-resistant Staphylococcus aureus.

  7. Differential Release of Lipoteichoic and Teichoic Acids from Streptococcus pneumoniae as a Result of Exposure to β-Lactam Antibiotics, Rifamycins, Trovafloxacin, and Quinupristin-Dalfopristin

    Stuertz, K; Schmidt, H.; Eiffert, H.; Schwartz, P.; Mäder, M.; Nau, R.

    1998-01-01

    The release of lipoteichoic acid (LTA) and teichoic acid (TA) from a Streptococcus pneumoniae type 3 strain during exposure to ceftriaxone, meropenem, rifampin, rifabutin, quinupristin-dalfopristin, and trovafloxacin in tryptic soy broth was monitored by a newly developed enzyme-linked immunosorbent assay. At a concentration of 10 μg/ml, a rapid and intense release of LTA and TA occurred during exposure to ceftriaxone (3,248 ± 1,688 ng/ml at 3 h and 3,827 ± 2,133 ng/ml at 12 h) and meropenem ...

  8. Familias de la proteína de superficie PspA de Streptococcus pneumoniae: Relación con serotipos y localización

    2010-01-01

    PspA, proteína de superficie de Streptococcus pneumoniae es un factor de virulencia, fuertemente inmunogénica y común a todos los serotipos. Aunque el gen que codifica para esta proteína presenta una marcada heterogeneidad en la región correspondiente al N-terminal, la PspA contiene epitopes conservados de manera tal que la inmunización genera protección contra neumococos pertenecientes a diversos tipos capsulares y con distintas PspA. A pesar del marcado polimorfismo del gen pspA es posible ...

  9. The Efflux Pump Inhibitor Reserpine Selects Multidrug-Resistant Streptococcus pneumoniae Strains That Overexpress the ABC Transporters PatA and PatB▿ †

    Garvey, Mark I.; Piddock, Laura J. V.

    2008-01-01

    One way to combat multidrug-resistant microorganisms is the use of efflux pump inhibitors (EPIs). Spontaneous mutants resistant to the EPI reserpine selected from Streptococcus pneumoniae NCTC 7465 and R6 at a frequency suggestive of a single mutational event were also multidrug resistant. No mutations in pmrA (which encodes the efflux protein PmrA) were detected, and the expression of pmrA was unaltered in all mutants. In the reserpine-resistant multidrug-resistant mutants, the overexpressio...

  10. Structure of the gene complementing uvr-402 in Streptococcus pneumoniae: homology with Escherichia coli uvrB and the homologous gene in Micrococcus luteus.

    Sicard, N.; Oreglia, J; Estevenon, A M

    1992-01-01

    The repair ability for UV-induced damage observed for Streptococcus pneumoniae proceeds through a system similar to the Uvr-dependent system in Escherichia coli. The DNA sequence of a gene complementing uvr-402, a mutation conferring UV sensitivity, was determined. Alignments of the deduced amino acid sequence revealed an extensive sequence homology of 55% with the UvrB protein of E. coli and 59% with the UvrB-homologous protein of Micrococcus luteus. Nucleotide-binding site consensus was obs...

  11. Meningoencephalitis caused by Streptococcus pneumoniae: a diagnostic and therapeutic challenge. Diagnosis with diffusion-weighted MRI leading to treatment with corticosteroids

    Jorens, Philippe G.; Demey, Hendrik E. [University Hospital of Antwerp, UZA, Department of Intensive Care Medicine, Edegem (Belgium); Parizel, Paul M. [University of Antwerp, Department of Radiology, Edegem (Belgium); Smets, Katrien [University of Antwerp, Department of Neurology, Edegem (Belgium); General Hospital AZ Middelares, Department of Neurology, Sint-Niklaas (Belgium); Jadoul, Kris [General Hospital AZ Middelares, Department of Neurology, Sint-Niklaas (Belgium); Verbeek, M.M.; Wevers, R.A. [University Hospital of Nijmegen, Laboratory of Paediatrics and Neurology, Nijmegen (Netherlands); Cras, Patrick [University of Antwerp, Department of Neurology, Edegem (Belgium)

    2005-10-01

    Streptococcus pneumoniae is a common cause of bacterial meningitis but only rarely causes other infections such as brain abscess, encephalitis, encephalomyelitis or meningoencephalitis. We report on three adult patients with meningoencephalitis caused by S. pneumoniae. In all three, CT and MRI revealed widespread brain lesions, suggesting extensive parenchymal injury. Diffusion-weighted MRI showed lesions with restricted diffusion, reflecting local areas of ischaemia with cytotoxic oedema secondary to an immunologically mediated necrotising vasculitis and thrombosis. High levels of markers of neuronal, glial and myelin damage were found in the cerebrospinal fluid. According to the literature, brain parenchyma lesions in adults with pneumococcal meningoencephalitis are often associated with death or severe neurological deficit. Our patients were treated with pulse doses of glucocorticoids: this resulted in dramatic clinical improvement and an excellent final neurological recovery. (orig.)

  12. Purification, crystallization and preliminary X-ray diffraction analysis of RafE, a sugar-binding lipoprotein from Streptococcus pneumoniae

    Paterson, Neil G., E-mail: neison@chem.gla.ac.uk; Riboldi-Tunnicliffe, Alan [Department of Chemistry and WestCHEM, Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom); Mitchell, Timothy J. [Division of Infection and Immunity (IBLS), Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom); Isaacs, Neil W. [Department of Chemistry and WestCHEM, Glasgow Biomedical Research Centre (GBRC), University of Glasgow, 120 University Place, Glasgow G12 8TA,Scotland (United Kingdom)

    2006-07-01

    The mature form of RafE has been expressed, purified and crystallized. X-ray diffraction data have been collected to 3.65 and 2.90 Å resolution from native and selenomethionine-derivative crystals, respectively. Streptococcus pneumoniae contains a large number of sugar-transport systems and the system responsible for raffinose uptake has recently been identified. The substrate-binding protein component of this system shares strong sequence homology with the multiple sugar metabolism substrate-binding protein MsmE from S. mutans and contains a lipoprotein-attachment site at cysteine residue 23. A truncated form (residues 24–419) of RafE from S. pneumoniae was cloned and overexpressed in Escherichia coli. Native and selenomethionine-labelled protein have been crystallized in the hexagonal space group P6{sub 1}22. Diffraction data have been successfully phased to 2.90 Å using Se SAD data and model building is in progress.

  13. Purification, crystallization and preliminary X-ray diffraction analysis of RafE, a sugar-binding lipoprotein from Streptococcus pneumoniae

    The mature form of RafE has been expressed, purified and crystallized. X-ray diffraction data have been collected to 3.65 and 2.90 Å resolution from native and selenomethionine-derivative crystals, respectively. Streptococcus pneumoniae contains a large number of sugar-transport systems and the system responsible for raffinose uptake has recently been identified. The substrate-binding protein component of this system shares strong sequence homology with the multiple sugar metabolism substrate-binding protein MsmE from S. mutans and contains a lipoprotein-attachment site at cysteine residue 23. A truncated form (residues 24–419) of RafE from S. pneumoniae was cloned and overexpressed in Escherichia coli. Native and selenomethionine-labelled protein have been crystallized in the hexagonal space group P6122. Diffraction data have been successfully phased to 2.90 Å using Se SAD data and model building is in progress

  14. Crystallization and preliminary X-ray crystallographic studies of DesR, a thermosensing response regulator in a two-component signalling system from Streptococcus pneumoniae

    The response regulator DesR from S. pneumoniae was cloned, expressed, purified and crystallized. A complete data set was collected to 1.7 Å resolution. The response regulator DesR, which activates the transcription of the des gene by binding to a regulatory region, is essential for controlling the fluidity of membrane phospholipids. DesR from Streptococcus pneumoniae was overexpressed in Escherichia coli. The protein was purified and crystallized for structural analysis. Diffraction data were collected to 1.7 Å resolution using synchrotron radiation and the crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 46.91, b = 71.38, c = 117.73 Å. Assuming the presence of a dimer in the asymmetric unit, this corresponds to a VM of 2.21 Å3 Da−1

  15. Delineation of Streptococcus dysgalactiae, its subspecies, and its clinical and phylogenetic relationship to Streptococcus pyogenes

    Jensen, Anders; Kilian, Mogens

    2011-01-01

    The close phylogenetic relationship of the important pathogen Streptococcus pneumoniae and several species of commensal streptococci, particularly Streptococcus mitis and Streptococcus pseudopneumoniae, and the recently demonstrated sharing of genes and phenotypic traits previously considered...

  16. Exit from competence for genetic transformation in Streptococcus pneumoniae is regulated at multiple levels.

    Weng, Liming; Piotrowski, Andrew; Morrison, Donald A

    2013-01-01

    Development of natural competence in S. pneumoniae entails coordinated expression of two sets of genes. Early gene expression depends on ComE, a response regulator activated by the pheromone CSP (Competence-Stimulating-Peptide). Subsequently, an early gene product (the alternative sigma factor ComX) activates expression of late genes, establishing the competent state. Expression of both sets of genes is transient, rapidly shut off by a mechanism that depends on the late gene, dprA. It has been thought that the rapid shutoff of late gene expression is the combined result of auto-inhibition of ComE and the instability of ComX. However, this explanation seems incomplete, because of evidence for ComX-dependent repressor(s) that might also be important for shutting off the response to CSP and identifying dprA as such a gene. We screened individual late gene mutants to investigate further the roles of ComX-dependent genes in competence termination. A ΔdprA mutant displayed a prolonged late gene expression pattern, whereas mutants lacking cbpD, cibABC, cglEFG, coiA, ssbB, celAB, cclA, cglABCD, cflAB, or radA, exhibited a wild-type temporal expression pattern. Thus, no other gene than dprA was found to be involved in shutoff. DprA limits the amounts of ComX and another early gene product, ComW, by restriction of early gene expression rather than by promoting proteolysis. To ask if DprA also affects late gene expression, we decoupled late gene expression from early gene regulation. Because DprA did not limit ComX activity under these conditions, we also conclude that ComX activity is limited by another mechanism not involving DprA. PMID:23717566

  17. The ecology of nasal colonization of Streptococcus pneumoniae, Haemophilus influenzae and Staphylococcus aureus: the role of competition and interactions with host's immune response

    Yates Andrew

    2010-02-01

    Full Text Available Abstract Background The first step in invasive disease caused by the normally commensal bacteria Streptococcus pneumoniae, Staphylococcus aureus and Haemophilus influenzae is their colonization of the nasal passages. For any population to colonize a new habitat it is necessary for it to be able to compete with the existing organisms and evade predation. In the case of colonization of these species the competition is between strains of the same and different species of bacteria and the predation is mediated by the host's immune response. Here, we use a neonatal rat model to explore these elements of the ecology of nasal colonization by these occasionally invasive bacteria. Results When neonatal rats are colonized by any one of these species the density of bacteria in the nasal passage rapidly reaches a steady-state density that is species-specific but independent of inoculum size. When novel populations of H. influenzae and S. pneumoniae are introduced into the nasal passages of neonatal rats with established populations of the same species, residents and invaders coexisted. However, this was not the case for S. aureus - the established population inhibited invasion of new S. aureus populations. In mixed-species introductions, S. aureus or S. pneumoniae facilitated the invasion of another H. influenzae population; for other pairs the interaction was antagonistic and immune-mediated. For example, under some conditions H. influenzae promoted an immune response which limited the invasion of S. pneumoniae. Conclusions Nasal colonization is a dynamic process with turnover of new strains and new species. These results suggest that multiple strains of either H. influenzae or S. pneumoniae can coexist; in contrast, S. aureus strains require a host to have no other S. aureus present to colonize. Levels of colonization (and hence the possible risk of invasive disease by H. influenzae are increased in hosts pre-colonized with either S. aureus or S

  18. On the analysis of the virulence nature of TIGR4 and R6 strains of Streptococcus pneumoniae using genome comparison tools

    R Jothi; K Manikandakumar; K Ganesan; S Parthasarathy

    2007-09-01

    Comparative genome sequence analysis is a powerful technique for gaining insights into any genome of interest. Streptococcus pneumoniae is a human pathogen, which causes life-threatening diseases, such as pneumoniae, bacteremia, meningitis, etc. After the whole genome of two strains of S. pneumoniae, the virulent TIGR4 and non-pathogenic R6 were sequenced; there is a hope that comparing the genomes will allow an identification of the genes responsible for its virulence and thus the development of treatment and control. Many antimicrobial drugs have diminished the risk from pneumococcal disease because of its multi-drug resistance nature. Several pneumococcal proteins are also being investigated, as virulence factors as potential vaccine or drug targets. Structural and biochemical studies of these pneumococcal virulence factors have facilitated the development of novel antibiotics or protein antigen-based vaccines for the treatment of pneumococcal disease. Here we describe the comparison between the genomes of two strains of S. pneumoniae with few existing genomics databases and tools available in the public domain websites. By comparing nucleotide and protein sequences of the two strains, we investigate the existing differences and similarities. Mainly we focus on the virulence factors and its encoding genes in TIGR4 and how do they differ from R6 strain.

  19. 探针荧光定量PCR在肺炎链球菌检测中的应用%Clinical application of real-time fluorescence quantitive PCR for detecting Streptococcus pneumoniae

    曹东林; 胡亮彬; 林茂锐; 王婷; 黄基伟; 田军章

    2014-01-01

    目的:建立探针荧光定量PCR检测肺炎链球菌的方法。方法针对肺炎链球菌种属特异性基因lytA,设计合成了特异引物和探针,研究引物和探针的灵敏度和特异性,确定循环阈值(cycle threshold,ct)的临界值(cut-off value)。将荧光定量PCR和细菌培养法进行比较,同时检测158份肺炎患者痰液标本加以验证。结果针对LytA基因所设计的引物和探针能灵敏地检出常见致病的血清型肺炎链球菌株,检测灵敏度为每个反应100个基因组DNA拷贝。通过荧光量PCR方法,35株肺炎链球菌中检测结果为阳性有34株,检测结果为阴性的有1株;15株非肺炎链球菌全部为阴性。通过荧光定量PCR方法,158份痰液标本中共检测出34份肺炎链球菌阳性,其中10份培养出相应的病原菌。肺炎链球菌阳性患者的白细胞数量和住院时间均显著高于阴性患者(P<0.05)。肺炎链球菌阳性患者住院时间均显著长于阴性患者(P<0.05)。结论探针荧光定量PCR方法能特异地检测肺炎链球菌,具有很高的灵敏度,能提高临床肺炎链球菌感染患者标本的阳性检出率。%Objective To establish an assay for the detection of Streptococcus pneumoniae by real-time fluorescence quantititive polymerase chain reaction (PCR).Methods Special primers and probe for the autolysin A (lytA)gene were designed.The sensitivity and specificity of primers and probe were studied,and cut-off of cycle threshold was assayed.158 clinical specimens were confirmed by real-time fluorescence quantitative PCR and bacterial culture method.Results Primer and probe design for LytA gene could sensitively detect serotype Streptococcus pneumoniae strains of common pathogenic,and the sensitivity was 100 copies.Among 35 strains of Streptococcus pneumoniae,34 cases were detected to be positive for Streptococcus pneumoniae by real-time fluorescence quantitative PCR,while 1 case was

  20. Outbreaks of Streptococcus pneumoniae carriage in day care cohorts in Finland – implications for elimination of transmission

    Auranen Kari

    2009-06-01

    Full Text Available Abstract Background Day care centre (DCC attendees play a central role in maintaining the circulation of Streptococcus pneumoniae (pneumococcus in the population. Exposure within families and within DCCs are the main risk factors for colonisation with pneumococcal serotypes in DCC attendees. Methods Transmission of serotype specific carriage was analysed with a continuous time event history model, based on longitudinal data from day care attendees and their family members. Rates of acquisition, conditional on exposure, were estimated in a Bayesian framework utilising latent processes of carriage. To ensure a correct level of exposure, non-participating day care attendees and their family members were included in the analysis. Posterior predictive simulations were used to quantify transmission patterns within day care cohorts, to estimate the basic reproduction number for pneumococcal carriage in a population of day care cohorts, and to assess the critical vaccine efficacy against carriage to eliminate pneumococcal transmission. Results The model, validated by posterior predictive sampling, was successful in capturing the strong temporal clustering of pneumococcal serotypes in the day care cohorts. In average 2.7 new outbreaks of pneumococcal carriage initiate in a day care cohort each month. While 39% of outbreaks were of size one, the mean outbreak size was 7.6 individuals and the mean length of an outbreak was 2.8 months. The role of families in creating and maintaining transmission was minimal, as only 10% of acquisitions in day care attendees were from family members. Considering a population of day care cohorts, a child-to-child basic reproduction number was estimated as 1.4 and the critical vaccine efficacy against acquisition of carriage as 0.3. Conclusion Pneumococcal transmission occurs in serotype specific outbreaks of carriage, driven by within-day-care transmission and between-serotype competition. An amplifying effect of the day

  1. The α-Tocopherol Form of Vitamin E Reverses Age-Associated Susceptibility to Streptococcus pneumoniae Lung Infection by Modulating Pulmonary Neutrophil Recruitment

    Ghanem, Elsa N. Bou; Clark, Stacie; Du, Xiaogang; Wu, Dayong; Camilli, Andrew; Leong, John M.; Meydani, Simin N.

    2016-01-01

    Streptococcus pneumoniae infections are an important cause of morbidity and mortality in older patients. Uncontrolled neutrophil-driven pulmonary inflammation exacerbates this disease. To test whether the α-tocopherol (α-Toc) form of vitamin E, a regulator of immunity, can modulate neutrophil responses as a preventive strategy to mitigate the age-associated decline in resistance to S. pneumoniae, young (4 mo) and old (22–24 mo) C57BL/6 mice were fed a diet containing 30-PPM (control) or 500-PPM (supplemented) α-Toc for 4 wk and intratracheally infected with S. pneumoniae. Aged mice fed a control diet were exquisitely more susceptible to S. pneumoniae than young mice. At 2 d postinfection, aged mice suffered 1000-fold higher pulmonary bacterial burden, 2.2-fold higher levels of neutrophil recruitment to the lung, and a 2.25-fold higher rate of lethal septicemia. Strikingly, α-Toc supplementation of aged mice resulted in a 1000-fold lower bacterial lung burden and full control of infection. This α-Toc–induced resistance to pneumococcal challenge was associated with a 2-fold fewer pulmonary neutrophils, a level comparable to S. pneumoniae–challenged, conventionally fed young mice. α-Toc directly inhibited neutrophil egress across epithelial cell monolayers in vitro in response to pneumococci or hepoxilin-A3, an eicosanoid required for pneumococcus-elicited neutrophil trans-epithelial migration. α-Toc altered expression of multiple epithelial and neutrophil adhesion molecules involved in migration, including CD55, CD47, CD18/CD11b, and ICAM-1. These findings suggest that α-Toc enhances resistance of aged mice to bacterial pneumonia by modulating the innate immune response, a finding that has potential clinical significance in combating infection in aged individuals through nutritional intervention. PMID:25512603

  2. 91株肺炎链球菌的血清型分布及耐药性分析%Drug resistance and serotypes of 91 strains of Streptococcus pneumoniae

    张波; 陈瑶; 刘智勇; 林钟劝; 夏宇; 和昱辰; 府伟灵

    2012-01-01

    目的 了解重庆地区肺炎链球菌临床分离株的血清型分布及药物敏感性.方法 采用荚膜肿胀试验进行肺炎链球菌血清学分型,并计算疫苗(PVC7、PVC11、PVC13)覆盖率;肉汤稀释法测定抗菌药物的最低抑菌浓度(MIC).结果 91株肺炎链球菌的临床分离患者年龄呈典型双峰分布,以<5岁婴幼儿与>50岁中老年人群为主,占51.7%、27.5%;90株肺炎链球菌共鉴定出20个血清型,1株未能血清分型,常见的肺炎链球菌血清型为19F、19A、6B,PVC13覆盖率为74.4%;91株肺炎链球菌均表现出较高的耐药率,在67株β-内酰胺类抗菌药物不敏感株(BLAs)中,青霉素不敏感菌株(PNSP)占53.8%.结论 重庆地区肺炎链球菌临床分离株以19F、19A、6B血清型为主,PVC13的预防作用更显著;肺炎链球菌耐药性高尤其是大多数菌株呈多药耐药趋势,临床应注意合理选择用药.%OBJECTIVE To investigate the antibiotics resistance and the prevalence of serotypes of Streptococcus pneumoniae isolated from the clinical patients in Chongqing. METHODS The capsular serotypes were identified by Quellung reaction and compared with the spectrum of the pneumococcal conjugate vaccine. Minimal inhibition concentrations (MICs) of the antibiotics were determined by the broth microdilution method. RESULTS The age profile of clinical patients infected by 91 strains of S. pneumoniae revealed a typical twin-peak distribution, with most patients being either <5 years old (51. 7%) or ≥50 years old (27. 5%). Among 91 clinical isolates, 90 were divided into 20 serotypes and 1 was unable to serotype. The most prevalent serotypes of S. pneumoniae isolates were 19F, 19A, 6B, and PVC13 was expected to cover 74. 4%. The majority of 91 isolates were resistant to common antibiotics. 53. 8% of 67 BLAs were penicillin non-susceptible S. pneumoniae. CONCLUSION The prevalent serotypes are 19F, 19A, and 6B in Chongqing, and PVC13 is an effective vaccine

  3. Progress of drug resistance of Streptococcus pneumoniae in China%国内耐药肺炎链球菌研究进展

    韦秋玲

    2013-01-01

      Streptococcus pneumoniae is an important pathogen that causes devastating infectious diseases,such as bacterial pneumonia,otitis media and meningitis in both developed and developing countries. WHO estimates that each year about 1.6 milion people have been infected with the bacteria and death ,Including 700000 ~ 1 milion for children under the age of five. And most people in developing countries.Due to the excessive use of antibiotics, the resistance of pneumococcal isolates to many of many of the commonly used antibiotics results in fewer effective antibiotics available for treatment. As the resistant S.pneumoniae is gradualy approaching what the “superbug”is, t Facing the serious situation ,Scientists streptococcus pneumoniae resistance to research and drug resistance gene detection and vaccine development, so as to seek the ideal solution. This article provides an overview of the current status of drug resistance of S.pneumoniae and Resistance genes related detection methods and control strategy is summarized in this paper. in China.%  肺炎链球菌可引起细菌性肺炎、中耳炎和脑膜炎等多种侵袭性疾病,是当今发达国家和发展中国家共有的一个重要病原,WHO估计每年约有160万人因感染此菌而死亡,其中70万~100万为5岁以下儿童。且多数生活在发展中国家。由于抗生素长期的过度使用,许多肺炎链球菌菌株能够同时耐受多种常用抗生素,耐药肺炎链球菌正在朝着“超级细菌”方向发展。面对这一严峻形势,科学家致力于肺炎链球菌耐药性研究及耐药基因的检测以及疫苗的开发,以期寻求理想的解决方法。本文对国内肺炎链球菌的流行现状及其耐药基因相关检测方法和防治策略进行综述。

  4. Vinpocetine inhibits Streptococcus pneumoniae-induced upregulation of mucin MUC5AC expression via induction of MKP-1 phosphatase in the pathogenesis of otitis media.

    Lee, Ji-Yun; Komatsu, Kensei; Lee, Byung-Cheol; Miyata, Masanori; O'Neill Bohn, Ashley; Xu, Haidong; Yan, Chen; Li, Jian-Dong

    2015-06-15

    Mucin overproduction is a hallmark of otitis media (OM). Streptococcus pneumoniae is one of the most common bacterial pathogens causing OM. Mucin MUC5AC plays an important role in mucociliary clearance of bacterial pathogens. However, if uncontrolled, excessive mucus contributes significantly to conductive hearing loss. Currently, there is a lack of effective therapeutic agents that suppress mucus overproduction. In this study, we show that a currently existing antistroke drug, vinpocetine, a derivative of the alkaloid vincamine, inhibited S. pneumoniae-induced mucin MUC5AC upregulation in cultured middle ear epithelial cells and in the middle ear of mice. Moreover, vinpocetine inhibited MUC5AC upregulation by inhibiting the MAPK ERK pathway in an MKP-1-dependent manner. Importantly, ototopical administration of vinpocetine postinfection inhibited MUC5AC expression and middle ear inflammation induced by S. pneumoniae and reduced hearing loss and pneumococcal loads in a well-established mouse model of OM. Thus, these studies identified vinpocetine as a potential therapeutic agent for inhibiting mucus production in the pathogenesis of OM. PMID:25972475

  5. The clinical distribution and drug resistance monitoring of Streptococcus pneumonia%肺炎链球菌的临床分布及耐药性监测

    梁培松; 孙各琴; 张秀明; 黄福达; 卢兰芬

    2015-01-01

    目的:了解该院肺炎链球菌的临床分布及监测其耐药变迁,为临床有效抗感染提供参考依据。方法用VITEK 2 Compact进行细菌鉴定和药敏数据分析,用WHONET5.3软件和SPSS13.0软件进行统计分析。结果2008~2013年共分离出588株肺炎链球菌,主要分布于重症监护病房(IC U ),其次为呼吸内科、普通儿科;主要来源于痰液标本,其次为咽拭子和血液标本。红霉素耐药率最高,其次为青霉素和复方磺胺甲噁唑;肺炎链球菌对左旋氧氟沙星、氧氟沙星、万古霉素、氯霉素、利奈唑烷仍然敏感。结论肺炎链球菌的耐药率不断上升,应重视细菌耐药性监测,根据药敏试验结果合理选用抗菌药物。%Objective To understand the clinical distribution and monitoring the change of resistance of Streptococcus pneumoni‐ae ,effective for clinical anti infection to provide reference .Methods Using VITEK 2 Compact to analyze the bacteria identification and drug sensitivity data ,and using WHONET5 .3 software and SPSS13 .0 software for statistical analysis .Results From 2008 to 2013 ,588 strains of Streptococcus pneumoniae were isolated ,mainly distributed in the intensive care unit (ICU) ,followed by respir‐atory department of internal medicine ,general pediatrics ;mainly from sputum samples ,followed by the throat swabs and blood samples .The highest resistant rate was erythromycin ,followed by penicillin and cotrimoxazole ;Streptococcus pneumoniae remains sensitive to ofloxacin ,levofloxacin ,vancomycin ,linezolid ,chloramphenicol .Conclusion The resistance rate of Streptococcus pneu‐moniae was rising ,and that great attention should be paid to the bacterial drug resistance so as to reasonably use a antibiotics based on the result of drug susceptibility testing .

  6. Nasopharyngeal colonization and invasive disease are enhanced by the cell wall hydrolases LytB and LytC of Streptococcus pneumoniae.

    Elisa Ramos-Sevillano

    Full Text Available BACKGROUND: Streptococcus pneumoniae is a common colonizer of the human nasopharynx and one of the major pathogens causing invasive disease worldwide. Dissection of the molecular pathways responsible for colonization, invasion, and evasion of the immune system will provide new targets for antimicrobial or vaccine therapies for this common pathogen. METHODOLOGY/PRINCIPAL FINDINGS: We have constructed mutants lacking the pneumococcal cell wall hydrolases (CWHs LytB and LytC to investigate the role of these proteins in different phases of the pneumococcal pathogenesis. Our results show that LytB and LytC are involved in the attachment of S. pneumoniae to human nasopharyngeal cells both in vitro and in vivo. The interaction of both proteins with phagocytic cells demonstrated that LytB and LytC act in concert avoiding pneumococcal phagocytosis mediated by neutrophils and alveolar macrophages. Furthermore, C3b deposition was increased on the lytC mutant confirming that LytC is involved in complement evasion. As a result, the lytC mutant showed a reduced ability to successfully cause pneumococcal pneumonia and sepsis. Bacterial mutants lacking both LytB and LytC showed a dramatically impaired attachment to nasopharyngeal cells as well as a marked degree of attenuation in a mouse model of colonization. In addition, C3b deposition and phagocytosis was more efficient for the double lytB lytC mutant and its virulence was greatly impaired in both systemic and pulmonary models of infection. CONCLUSIONS/SIGNIFICANCE: This study confirms that the CWHs LytB and LytC of S. pneumoniae are essential virulence factors involved in the colonization of the nasopharynx and in the progress of invasive disease by avoiding host immunity.

  7. 国内耐药肺炎链球菌的流行现状%Current status of drug resistance of Streptococcus pneumoniae in China

    姚开虎; 张敬仁

    2011-01-01

    Streptococcus pneumoniae is an important pathogen that causes devastating infectious diseases, such as bacterial pneumonia, otitis media and meningitis in both developed and developing countries. Due to the excessive use of antibiotics, the re sistance of pneumococcal isolates to many of the commonly used antibiotics results in fewer effective antibiotics available for treat ment. As the resistant S. pneumoniae is gradually approaching what the “superbug” is, the emergent situation demands our full at tention and efforts in seeking short-and long-term solutions. This article provides an overview of the current status of drug resis tance of S. pneumoniae and the related research in China.%肺炎链球菌可引起细菌性肺炎、中耳炎和脑膜炎等疾病,是当今发达国家和发展中国家共有的一个重要病原.由于抗生素长期的过度使用,许多肺炎链球菌菌株能够同时耐受多种常用的抗生素,使得临床可应用的有效抗生素越来越少.耐药肺炎链球菌正在朝着"超级细菌"方向发展.这种现状值得我们关注并寻求近期和长期的解决方法.本文对国内肺炎链球菌耐药现状及相关研究进行综述.

  8. Bacteremia with Streptococcus pneumoniae

    Christensen, J S; Jensen, T G; Kolmos, H J;

    2012-01-01

    severe sepsis, and 11 (3 %) were in septic shock. Overall, the 30-day mortality was 16 %. Mortality increased with the severity of sepsis. There was no association between the focal diagnosis of SPB or the number of diagnoses and mortality. Nosocomial infection, male sex, increasing age, and increasing...

  9. 儿童鼻部携带肺炎链球菌的研究%Nasal carriage of Streptococcus pneumoniae among children in Beijing

    李洁; 袁林; 俞桑洁; 杨永弘

    2001-01-01

    Objective To investigate the antimicrobial susceptibility of Streptococcus pneumoniae carried in the nose among children in Beijing and the distribution of serotypes,and to analyze the risk factors for nasal carriage of penicillin non-susceptible S.pneumoniae.Methods A disk diffusion test was applied to detect the antimicrobial susceptibilities of S.pneumoniae to erythromycin,trimethoprim-sulfamethoxazole,chloramphenicol and tetracycline.The E test was applied to determine the minimal inhibitory concentrations of penicillin,cefuroxime,cefotaxime,augmentin and imipenem.S.pneumoniae isolates were serotyped by the Quellung reaction.Results S.pneumoniae that was resistant to penicillin or cefuroxime was not found,but S.pneumoniae intermediate resistant to penicillin and cefuroxime accounted for 8.2% and 2.1%,respectively.All of the isolates were susceptible to cefotaxime,augmentin and imipenem.S.pneumonia that was resistant to erythromycin,trimethoprim-sulfamethoxazole and tetracycline were extremely numerous,accounting for 72%,70% and 79%,respectively.Five serotypes(19,6,14,23,17)accounted for 54.7%,and nontypables accounted for 20.6% of all the S.pneumoniae.Previous history of otitis media was a risk factor we found for nasal carriage of penicillin non-susceptible S.pneumoniae.Conclusions Continued surveillance of the antimicrobial susceptibilities of S.pneumoniae is necessary.A larger scale investigation is needed to identify if the 7 or 9-valent conjugate pneumococcal vaccine is appropriate for Chinese children.%目的了解北京地区儿童鼻部携带的肺炎链球菌对抗生素的敏感性以及血清型分布,分析鼻部携带青霉素非敏感肺炎链球菌的危险因素.方法用纸片扩散法检测肺炎链球菌对红霉素,复方新诺明,氯霉素和四环素的敏感性;E-试验确定青霉素,头孢呋新,头孢噻肟,安灭菌和亚胺培南的最小抑菌浓度;Quellung反应确定肺炎链球菌的血清型.结果未发现对青霉素和头

  10. Evaluación y evidencia para la vacunación contra rotavirus y vacunación contra Streptococcus pneumonia en Costa Rica

    Ricardo Morales Vargas

    2006-12-01

    Full Text Available Se evaluaron y compararon dos posibles intervenciones en salud pública para ser aplicadas en Costa Rica: a vacunación contra el rotavirus y b vacunación contra el Streptococcus pneumonia. Se utilizó la técnica de revisión de la literatura impresa y electrónica usando como marco de referencia para la evaluación las recomendaciones de la International Network of Agencies of Health Care Technology Assessment -(INHATA y las recomendaciones de la OPS/OMS sobre evaluación de tecnologías sanitarias para países en desarrollo. Se utilizaron los criterios de seguridad, eficacia, efectividad, utilidad y costo-beneficio de ambas intervenciones, así como su impacto organizacional, ético y social. Se escogieron como grupos en riesgo las personas menores de 5 años y los mayores de 65 años. Se emplearon datos económicos publicados por otros autores sobre costos sanitarios y los precios de mercado para las vacunas. Considerando los aspectos anteriores, así como la calidad y amplitud de la literatura sobre ambas intervenciones, se concluye que la intervención contra el rotavirus podría ser una intervención de mayor impacto en cuanto a número de casos evitables en Costa Rica, por lo que se recomienda realizar estudios nacionales de Fase III para evaluar su efectividad. Si bien la intervención contra el Streptococcus pneumonia (C/B =2,4 es una intervención más favorable económicamente que la del rotavirus (C/B =2,7 en términos de costo/beneficio en el mediano y largo plazo, esta conclusión se revertiría si sólo se consideran los costos anuales luego del primer año. Resultados de la evaluación global, utilizando los criterios anteriores, propuesta por el autor, favorece la intervención ante el rotavirus.Two posible health interventions were evaluated and compared for application in Costa Rica: a vaccination against Rotavirus, and b vaccination against Streptococcus pneumonia. Analysis of existing printed and electronic literature was

  11. Resistance and serotype distribution of Streptococcus pneumoniae among adults and children in China%我国成人和儿童中分离的肺炎链球菌的耐药性与血清型研究

    肖素坤; 赵春江; 刘春林; 王辉

    2010-01-01

    Objective To investigate the serogroups/types distribution and antimicrobial susceptibility of clinical Streptococcus pneumoniae isolates of different serogroups/types in both children and adults in China, and to explore the significance of vaccines in preventing pneumococcal infections and control of epidemic Streptococcus pneumoniae. Methods A total of 580 consecutive and non-repetitive Streptococcuspneumoniae isolates were collected from 13 hospitals between 2005 and 2008. Agar dilution method was used to determine the minimal inhibitory concentrations ( MIC) of 11 antibacterial agents. Serotyping was performed by latex agglutination test and the Quellung reaction test. Results The most prevalent serogroups/types in 362 isolates from adults were 19F (55, 15.2%), 19A (46, 12.7%), 3 (44, 12.2%), 23F (24, 6.6%), 15 (23, 6.4%), and 17 (11, 3.0%), while in the 218 isolates from children, 19F (71, 32.6%), 19A (31, 14.2%), 23F (13, 6.0%), 15 (12, 5.5%), 14 (11, 5. 0% ) , and 6B ( 10, 4. 6% ) were the most prevalent Resistance to (J-lactams was related to the serotypes. 19F and 19A were more resistant toβ-lactams than the other serotypes. The prevalence of Penicillin Intermediate Streptococcus pneumoniae increased from 7. 4% in 2005 to 24. 9% in 2008. The coverage of pnuemococcal conjugate vaccine (PCV)-7, PCV-10 and PCV-13 among all age groups was 35.5% (206/580), 38.7% (224/580) and 61. 8% (358/580), respectively. The coverage of PCV-7, PCV-10 and PCV-13 among children under 5 years was 55. 7% (78/140), 58. 6% (82/140), and 77. 9% (109/140) respectively. Conclusion Penicillin intermediate isolates were on the rise with years. PCV-7, PCV-10 and PCV-13 vaccines showed a higher coverage in children than in adults.%目的 研究自我国成人和儿童中分离的肺炎链球菌的血清分型,探索不同血清型对常用抗菌药物的敏感度,评估肺炎链球菌疫苗在预防肺炎链球菌感染及控制肺炎链球菌流行传播中的价值.方法 收集2005

  12. Crystallization and preliminary crystallographic analysis of the bacterial capsule assembly-regulating tyrosine phosphatases Wzb of Escherichia coli and Cps4B of Streptococcus pneumoniae

    The crystallization is reported of two bacterial tyrosine phosphatases which belong to different enzyme families despite their ability to catalyse identical reactions. Bacterial tyrosine kinases and their cognate phosphatases are key players in the regulation of capsule assembly and thus are important virulence determinants of these bacteria. Examples of the kinase/phosphatase pairing are found in Gram-negative bacteria such as Escherichia coli (Wzc and Wzb) and in Gram-positive bacteria such as Streptococcus pneumoniae (CpsCD and CpsB). Although Wzb and Cps4B are both predicted to dephosphorylate the C-terminal tyrosine cluster of their cognate tyrosine kinase, they appear on the basis of protein sequence to belong to quite different enzyme classes. Recombinant purified proteins Cps4B of S. pneumoniae TIGR4 and Wzb of E. coli K-30 have been crystallized. Wzb crystals belonged to space-group family P3x21 and diffracted to 2.7 Å resolution. Crystal form I of Cps4B belonged to space-group family P4x212 and diffracted to 2.8 Å resolution; crystal form II belonged to space group P212121 and diffracted to 1.9 Å resolution

  13. Biological and Epidemiological Features of Antibiotic-Resistant Streptococcus pneumoniae in Pre- and Post-Conjugate Vaccine Eras: a United States Perspective.

    Kim, Lindsay; McGee, Lesley; Tomczyk, Sara; Beall, Bernard

    2016-07-01

    Streptococcus pneumoniae inflicts a huge disease burden as the leading cause of community-acquired pneumonia and meningitis. Soon after mainstream antibiotic usage, multiresistant pneumococcal clones emerged and disseminated worldwide. Resistant clones are generated through adaptation to antibiotic pressures imposed while naturally residing within the human upper respiratory tract. Here, a huge array of related commensal streptococcal strains transfers core genomic and accessory resistance determinants to the highly transformable pneumococcus. β-Lactam resistance is the hallmark of pneumococcal adaptability, requiring multiple independent recombination events that are traceable to nonpneumococcal origins and stably perpetuated in multiresistant clonal complexes. Pneumococcal strains with elevated MICs of β-lactams are most often resistant to additional antibiotics. Basic underlying mechanisms of most pneumococcal resistances have been identified, although new insights that increase our understanding are continually provided. Although all pneumococcal infections can be successfully treated with antibiotics, the available choices are limited for some strains. Invasive pneumococcal disease data compiled during 1998 to 2013 through the population-based Active Bacterial Core surveillance program (U.S. population base of 30,600,000) demonstrate that targeting prevalent capsular serotypes with conjugate vaccines (7-valent and 13-valent vaccines implemented in 2000 and 2010, respectively) is extremely effective in reducing resistant infections. Nonetheless, resistant non-vaccine-serotype clones continue to emerge and expand. PMID:27076637

  14. Comparison of the serum-supplemented Todd-Hewitt and the new Haemophilus test media for broth microdilution susceptibility testing of Streptococcus pneumoniae.

    Roger, M; Lapointe, J R

    1996-06-01

    Horse serum-supplemented Todd-Hewitt broth (STH) in use at Hôpital Ste-Justine for the last 12 years was compared to the recently proposed Haemophilus test medium (HTM), for broth microdilution susceptibility testing of Streptococcus pneumoniae. One hundred and twenty S. pneumoniae isolates from pediatric clinical specimens were used in this study. In general, the minimum inhibitory concentrations (MICs) in STH for 15 antimicrobial agents were quite comparable to those determined in HTM but tended to be higher. Drugs which generated MICs within +/- 1 log2 concentration differences in both media included penicillin, ampicillin, oxacillin, cefuroxime, cefotaxime, cefixime, clindamycin, chloramphenicol, trimethoprim-sulfamethoxazole, rifampin, ciprofloxacin and vancomycin. Cefaclor and tetracycline MICs tended to be > or = 2 log2 dilutions higher with STH for most of the isolates tested, while erythromycin MICs were often 2 log2 dilutions lower with STH than with HTM. Despite some differences in MICs noted above, few very major (0.4%), major (0.2%) and minor interpretive category errors (4.4%) were observed. The visual reading of the MICs for most of the 120 clinical isolates tested was generally easier in STH which was superior in supporting best the bacterial growth as detected by spectrophotometry. The risk of false susceptibility is thus decreased by using STH rather than HTM; furthermore, STH is free of the technical problems of the lysed horse blood Mueller-Hinton (LHB-MH) recommended by the NCCLS. PMID:8808713

  15. New insights into histidine triad proteins: solution structure of a Streptococcus pneumoniae PhtD domain and zinc transfer to AdcAII.

    Beate Bersch

    Full Text Available Zinc (Zn(2+ homeostasis is critical for pathogen host colonization and invasion. Polyhistidine triad (Pht proteins, located at the surface of various streptococci, have been proposed to be involved in Zn(2+ homeostasis. The phtD gene, coding for a Zn(2+-binding protein, is organized in an operon with adcAII coding for the extracellular part of a Zn(2+ transporter. In the present work, we investigate the relationship between PhtD and AdcAII using biochemical and structural biology approaches. Immuno-precipitation experiments on purified membranes of Streptococcus pneumoniae (S. pneumoniae demonstrate that native PhtD and AdcAII interact in vivo confirming our previous in vitro observations. NMR was used to demonstrate Zn(2+ transfer from the Zn(2+-bound form of a 137 amino acid N-terminal domain of PhtD (t-PhtD to AdcAII. The high resolution NMR structure of t-PhtD shows that Zn(2+ is bound in a tetrahedral site by histidines 83, 86, and 88 as well as by glutamate 63. Comparison of the NMR parameters measured for apo- and Zn(2+-t-PhtD shows that the loss of Zn(2+ leads to a diminished helical propensity at the C-terminus and increases the local dynamics and overall molecular volume. Structural comparison with the crystal structure of a 55-long fragment of PhtA suggests that Pht proteins are built from short repetitive units formed by three β-strands containing the conserved HxxHxH motif. Taken together, these results support a role for S. pneumoniae PhtD as a Zn(2+ scavenger for later release to the surface transporter AdcAII, leading to Zn(2+ uptake.

  16. Seasonal Variation of Escherichia coli, Staphylococcus aureus, and Streptococcus pneumoniae Bacteremia According to Acquisition and Patient Characteristics: A Population-Based Study.

    Gradel, Kim Oren; Nielsen, Stig Lønberg; Pedersen, Court; Knudsen, Jenny Dahl; Østergaard, Christian; Arpi, Magnus; Jensen, Thøger Gorm; Kolmos, Hans Jørn; Søgaard, Mette; Lassen, Annmarie Touborg; Schønheyder, Henrik Carl

    2016-08-01

    OBJECTIVE Seasonal variation is a characteristic of many infectious diseases, but relatively little is known about determinants thereof. We studied the impact of place of acquisition and patient characteristics on seasonal variation of bacteremia caused by the 3 most common pathogens. DESIGN Seasonal variation analysis. METHODS In 3 Danish health regions (2.3 million total inhabitants), patients with bacteremia were identified from 2000 through 2011 using information from laboratory information systems. Analyses were confined to Escherichia coli, Staphylococcus aureus, and Streptococcus pneumoniae. Additional data were obtained from the Danish National Hospital Registry for the construction of admission histories and calculation of the Charlson comorbidity index (CCI). Bacteremias were categorized as community acquired, healthcare associated (HCA), and hospital acquired. We defined multiple subgroups by combining the following characteristics: species, acquisition, age group, gender, CCI level, and location of infection. Assuming a sinusoidal model, seasonal variation was assessed by the peak-to-trough (PTT) ratio with a 95% confidence interval (CI). RESULTS In total, we included 16,006 E. coli, 6,924 S. aureus, and 4,884 S. pneumoniae bacteremia cases. For E. coli, the seasonal variation was highest for community-acquired cases (PTT ratio, 1.24; 95% CI, 1.17-1.32), was diminished for HCA (PTT ratio, 1.14; 95% CI, 1.04-1.25), and was missing for hospital-acquired cases. No seasonal variation was observed for S. aureus. S. pneumoniae showed high seasonal variation, which did not differ according to acquisition (overall PTT ratio, 3.42; 95% CI, 3.10-3.83). CONCLUSIONS Seasonal variation was mainly related to the species although the place of acquisition was important for E. coli. Infect Control Hosp Epidemiol 2016;37:946-953. PMID:27142942

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

    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....... meningitidis in cells expressing TLR2, -4, or -9. It is interesting that the ability of S. pneumoniae and N. meningitidis to activate TLR9 was severely attenuated when bacteria had been heat-inactivated prior to stimulation of the cells. In human peripheral blood mononuclear cells, we blocked TLR2, -4, or -9...... infection with live pathogens may lead to activation of PRR not targeted by inactivated bacteria....

  18. The Vitamin B6 Biosynthesis Pathway in Streptococcus pneumoniae Is Controlled by Pyridoxal 5′-Phosphate and the Transcription Factor PdxR and Has an Impact on Ear Infection

    El Qaidi, Samir; Yang, Jun; Zhang, Jing-Ren; Metzger, Dennis W.; Bai, Guangchun

    2013-01-01

    Vitamin B6 is an essential cofactor for a large number of enzymes in both prokaryotes and eukaryotes. In this study, we characterized the pyridoxal 5′-phosphate (PLP) biosynthesis pathway in Streptococcus pneumoniae. Our results revealed that S. pneumoniae possesses a de novo vitamin B6 biosynthesis pathway encoded by the pdxST genes. Purified PdxS functionally displayed as PLP synthase, whereas PdxT exhibited glutaminase activity in vitro. Deletion of pdxS, but not pdxT, resulted in a vitami...

  19. Evaluation of oxacillin disk test for screening Streptococcus pneumoniae by penicillin Etest method%青霉素 Etest 法评价肺炎链球菌苯唑西林的纸片筛选试验

    罗湘蓉; 李成瑶; 袁军; 李红凌; 金婷婷; 胡方芳

    2014-01-01

    Objective To evaluate the oxacillin disk screening test for screening Streptococcus pneumoniae by the penicillin Etest method.Methods 96 clinically isolated non-meningitis strains of Streptococcus pneumoniae were collected.The sensitivity and spe-cificity of the oxacillin disc screening test was evaluated by the penicillin Etest method as a standard method for detecting the peni-cillin susceptibility.Results Among 96 non-meningitis strains of Streptococcus pneumoniae,the penicillin-susceptible Streptococcus pneumoniae(PSSP)strains detected by the penicillin Etest method accounted for 96.9%(93/96),the penicillin intermediate Strep-tococcus pneumoniae(PISP)strains accounted for 3.1%(3/96)and no penicillin resistant Streptococcus pneumoniae(PRSP)strain was found.But 16 PSSP strains were detected by the oxacillin disc screening test with the sensitivity of 17.2% and the specificity of 100.0%,respectively.The difference between the oxacillin disc screening test and the penicillin Etest method was statistically sig-nificant(χ2 =77,P <0.01 ).Conclusion The oxacillin disc screening test has the low sensitivity for preliminarily screening non-meningitis strains of Streptococcus pneumoniae.Most of Streptococcus pneumoniae must be detected by the minimum inhibitory concentration(MIC)methods such as the penicillin Etest method.%目的:青霉素 Etest 法评价肺炎链球菌苯唑西林的纸片筛选试验。方法对临床分离的96株肺炎链球菌非脑膜炎株,用 Etest 作为测定青霉素敏感性的标准方法,评价苯唑西林的纸片筛选试验的灵敏度和特异性。结果96株肺炎链球菌非脑膜炎株,用 Etest 测定青霉素敏感株(PSSP)为96.9%(93/96),青霉素中度敏感株(PISP)为3.1%(3/96),未发现青霉素耐药株(PRSP)。苯唑西林纸片法初筛出16株 PSSP,灵敏度为17.2%,特异性为100.0%。苯唑西林纸片法与 Etest 方法比较,差异有统计学意义(χ2=77

  20. 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

    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

  1. 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

    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...

  2. Toll-Like Receptor 3/TRIF-Dependent IL-12p70 Secretion Mediated by Streptococcus pneumoniae RNA and Its Priming by Influenza A Virus Coinfection in Human Dendritic Cells

    Spelmink, Laura; Sender, Vicky; Hentrich, Karina; Kuri, Thomas; Plant, Laura; Henriques-Normark, Birgitta

    2016-01-01

    ABSTRACT A functional immune response is crucial to prevent and limit infections with Streptococcus pneumoniae. Dendritic cells (DCs) play a central role in orchestrating the adaptive and innate immune responses by communicating with other cell types via antigen presentation and secretion of cytokines. In this study, we set out to understand how pneumococci activate human monocyte-derived DCs to produce interleukin-12 (IL-12) p70, an important cytokine during pneumococcal infections. We show ...

  3. Identification of Multiple Substrates of the StkP Ser/Thr Protein Kinase in Streptococcus pneumoniae

    Nováková, Linda; Bezoušková, Silvia; Pompach, Petr; Přenosilová, Lenka; Weiser, Jaroslav; Branny, Pavel

    2010-01-01

    Roč. 192, č. 14 (2010), s. 3629-3638. ISSN 0021-9193 R&D Projects: GA ČR GA204/08/0783; GA AV ČR IAA600200801; GA ČR GP204/07/P082 Institutional research plan: CEZ:AV0Z50200510 Keywords : GROUP-B STREPTOCOCCUS * EUKARYOTIC-TYPE * SERINE/THREONINE KINASE Subject RIV: EE - Microbiology, Virology Impact factor: 3.726, year: 2010

  4. Interaction between influenza virus and Streptococcus pneumoniae%流感病毒与肺炎链球菌相互作用的机制

    万天华

    2011-01-01

    流感死亡病例多与继发细菌感染,尤其是肺炎链球菌感染有关.病毒感染损伤正常保护性上皮层,引起小气道阻塞等呼吸道功能改变,有利于细菌定植和繁殖.流感病毒感染后的免疫反应也有利于继发细菌感染.细菌可增强病毒的致病力,细菌产生的蛋白酶可以裂解活化病毒血凝素的糖蛋白,使其获得感染力.细菌感染与原发流感病毒感染同时存在的患者症状严重,与抗生素治疗无效及细菌增强了原发感染病毒的毒力有关.认识流感病毒和细菌相互作用机制,将有助于制定预防和治疗重症流感病例的有效措施.%The mortality cases caused by influenza virus are offen associated with the secondary bacterial infection, Streptococcus pneumoniae infection in particularly. The mechanisms underlying the interaction between the virus and the bacteria are unclear at present. The damage of normal protective epithelial layer and changes in airway function caused by influenza virus provides increased numbers of attachment sites and culture spaces for the bacteria. The effect of influenza on immune system is a further opportunity for subsequent bacterial infection. The synergistic effect between influenza virus and Streptococcus pneumoniae might not be unidirectional. The bacteria caused the secondary infection could enhance the pathogenicity of the virus. The bacterium-derived proteases could cleave hemagglutinin to its active components that are required for infectivity of the virus. The effect of the bacteria on the virus could enhance the lethality of the primary viral disease, which can explain why antibiotic therapy was less successful to treat the influenza patient with subsequent bacteria infection. Effective strategies to prevent and treat the severe diseases could be suggested based on the knowledge of interaction between virus and bacteria.

  5. Pneumococcal Serotype 19F Conjugate Vaccine Induces Cross-Protective Immunity to Serotype 19A in a Murine Pneumococcal Pneumonia Model

    Jakobsen, Håvard; Sigurdsson, Viktor D.; Sigurdardottir, Sigurveig; Schulz, Dominique; Jonsdottir, Ingileif

    2003-01-01

    Immunization with a pneumococcal conjugate vaccine (PNC) containing serotype 19F induces cross-reactive antibodies to 19A in mice and human infants. Active immunization with PNC and passive immunization with serum samples from infants vaccinated with PNC containing serotype 19F, but not serotype 19A, protected against lung infection caused by both serotypes in a murine model.

  6. Effect of Ampicillin on the kinetics of colonization of Streptococcus pneumoniae and Lactobacillus fermentum in the respiratory tract of mice

    Silva Clara

    2004-10-01

    Full Text Available Abstract Ampicillin was selected to further study the effect of this antibiotic on the colonization capability of S. pneumoniae and L. fermentum intranasally inoculated in a mice experimental model. The sensitivity of S. pneumoniae and L. fermentum to antibiotics was evaluated by different "in vitro" techniques. The results showed that both microorganisms have a typical pattern of sensitivity to antibiotics in these assays. The "in vivo" experiments showed that the treatment with Ampicillin increased the number of lactobacilli and neumococci in the groups of mice treated only with one of the microorganisms. In those mice treated with Lactobacillus, challenged later with neumococci and treated with Ampicillin, the pathogen in lung decreased on the 4th day, disappearing completely after on. The histological studies showed that the antibiotic treatment decreased the inflammatory response produced by the pathogen at the lung and trachea levels.

  7. PspA Family Distribution, Antimicrobial Resistance and Serotype of Streptococcus pneumoniae Isolated from Upper Respiratory Tract Infections in Japan

    Muneki Hotomi; Akihisa Togawa; Masamitsu Kono; Yorihiko Ikeda; Shin Takei; Hollingshead, Susan K.; Briles, David E.; Kenji Suzuki; Noboru Yamanaka

    2013-01-01

    BACKGROUND: The protection against pneumococcal infections provided by currently available pneumococcal polysaccharide conjugate vaccines are restricted to the limited number of the serotypes included in the vaccine. In the present study, we evaluated the distribution of the pneumococcal capsular type and surface protein A (PspA) family of pneumococcal isolates from upper respiratory tract infections in Japan. METHODS: A total of 251 S. pneumoniae isolates from patients seeking treatment for ...

  8. Genome-wide association study of IgG1 responses to the choline-binding protein PspC of Streptococcus pneumoniae.

    Anderson, D; Fakiola, M; Hales, B J; Pennell, C E; Thomas, W R; Blackwell, J M

    2015-01-01

    Streptococcus pneumoniae causes invasive pneumococcal disease. Delayed development of antibodies to S. pneumoniae in infancy is associated with the development of atopy and asthma. Pneumococcal surface protein C (PspC) is a vaccine candidate and variation in its choline-binding region is associated with invasive disease. This study examined 523 060 single-nucleotide polymorphisms in The Western Australian Pregnancy Cohort (Raine) Study to find loci influencing immunoglobulin G1 (IgG1) responses to PspC measured at age 14 years (n=1152). Genome-wide significance (top SNP rs9275596; P=3.1 × 10(-14)) was only observed at human leucocyte antigen (HLA). Imputed HLA amino-acid polymorphisms showed the strongest associations at positions DRB1 47 (P=3.2 × 10(-11)), 13SRG (P=9.8 × 10(-10)) and 11SP (P=9.8 × 10(-10)), and at DQA1 34 (P=6.4 × 10(-10)), DQB1 167R (P=9.3 × 10(-6)) and HLA-B 95 W (P=1.2 × 10(-9)). Conditional analyses showed independent contributions from DRB1 47 and DQB1 167R to the signal at rs9275596, supported by an omnibus test showing a strong signal for the haplotype DRB1_47_DQB1_167 (P=9.02 × 10(-15)). In silico analysis showed that DRB1 four-digit allele groups defined by DRB1 47F bind to a greater complexity of core 9-mer epitopes compared with DRB1 47Y, especially across repeats in the C-term choline-binding region. Consequent differences in CD4 T-cell help for IgG1 to PspC could have implications for vaccine design. Further analysis in other cohorts is merited. PMID:25928883

  9. The highly conserved serine threonine kinase StkP of Streptococcus pneumoniae contributes to penicillin susceptibility independently from genes encoding penicillin-binding proteins

    Dias Ricardo

    2009-06-01

    Full Text Available Abstract Background The serine/threonine kinase StkP of Streptococcus pneumoniae is a major virulence factor in the mouse model of infection. StkP is a modular protein with a N-terminal kinase domain a C-terminal PASTA domain carrying the signature of penicillin-binding protein (PBP and prokaryotic serine threonine kinase. In laboratory cultures, one target of StkP is the phosphoglucosamine mutase GlmM involved in the first steps of peptidoglycan biosynthesis. In order to further elucidate the importance of StkP in S. pneumoniae, its role in resistance to β-lactams has been assessed by mutational analysis in laboratory cultures and its genetic conservation has been investigated in isolates from infected sites (virulent, asymptomatic carriers, susceptible and non-susceptible to β-lactams. Results Deletion replacement mutation in stkP conferred hypersensitivity to penicillin G and was epistatic on mutations in PBP2X, PBP2B and PBP1A from the resistant 9V clinical isolate URA1258. Genetic analysis of 55 clinical isolates identified 11 StkP alleles differing from the reference R6 allele. None relevant mutation in the kinase or the PASTA domains were found to account for susceptibility of the isolates. Rather the minimal inhibitory concentration (MIC values of the strains appeared to be determined by their PBP alleles. Conclusion The results of genetic dissection analysis in lab strain Cp1015 reveal that StkP is involved in the bacterial response to penicillin and is epistatic on mutations PBP 2B, 2X and 1A. However analysis of the clinical isolates did not allow us to find the StkP alleles putatively involved in determining the virulence or the resistance level of a given strain, suggesting a strong conservation of StkP in clinical isolates.

  10. Matrix metalloproteinase-9 deficiency impairs host defense mechanisms against Streptococcus pneumoniae in a mouse model of bacterial meningitis.

    Böttcher, Tobias; Spreer, Annette; Azeh, Ivo; Nau, Roland; Gerber, Joachim

    2003-03-01

    Matrix metalloproteinase-9 (MMP-9) appears to contribute to blood-brain barrier damage and neuronal injury in bacterial meningitis. To further explore the function of MMP-9 in meningeal inflammation, we injected 10(4) colony forming units (CFU) of a Streptoccocus pneumoniae type 3 strain into the right forebrain of MMP-9 deficient mice (MMP-9(-/-), n=16) and wild-type controls (129 x B6, n=15). The clinical course of the disease, leukocyte recruitment into the subarachnoid space and bacterial titers in the brain did not differ. Yet, clearance of the bacteria from blood (log CFU/ml 4.7 [3.8/5.4] vs. 3.6 [3.0/4.0]; P=0.005) and spleen homogenates (log CFU/ml 5.3 [4.8/5.5] vs. 4.0 [2.8/4.7]; P=0.01) was reduced in MMP-9 deficient mice. A reduced systemic bacterial clearance of MMP-9(-/-) mice was confirmed in experimental S. pneumoniae peritonitis/sepsis. This implies a compromised systemic, but not intracerebral host response against S. pneumoniae in MMP-9 deficiency. PMID:12581831

  11. Susceptibilidad antimicrobiana del Streptococcus pneumoniae determinando la concentración inhibitoria mínima, 1999

    Sara Morales de Santa Gadea

    2001-01-01

    Full Text Available Objetivo: Determinar la concentración inhibitoria mínima de S. pneumoniae frente a los antibióticos recomendados por OPS/OMS en el tratamiento de infecciones respiratorias agudas. Materiales y métodos: Estudio descriptivo de cultivos de S. pneumoniae aislados de niños menores de 5 años de edad con diagnóstico de neumonía y meningitis procedentes de siete hospitales del país. Se realizó el método de microdilución en placa para penicilina, cloranfenicol y cotrimoxazol siguiendo las pautas del Comité, Nacional de Estándares de Laboratorios Clínicos (NCCLS. Resultados: De las 30 muestras aisladas (18 de sangre y 12 de líquido cefalorraquídeo se observó susceptibilidad disminuida a penicilina en 8 (26.7%, sensibilidad intermedia en 4 (13.3% y resistencia alta en 4 (13.3%. En 21 aislamientos (70.0% se observó resistencia a cotrimoxazol (sulfametoxazol+trimetoprim y en 10 (33.0% a cloranfenicol. En 12 aislamientos (40.0% se observó resistencia a dos antimicrobianos y multirresistencia en 2 (6.7%. Conclusiones: Se logró determinar la presencia de cepas de S. pneumoniae resistentes a penicilina y a otros antimicrobianos, lo que nos obliga a mantener su vigilancia epidemiológica.

  12. Comparative in-vitro efficacy of fluoroquinolones against Streptococcus pneumoniae recovered from bacterial keratitis as determined by E-test

    Ramakrishnan R

    2010-04-01

    Full Text Available Background and Objectives: The advent of new fluoroquinolones has drawn the attention for reliable methods on the in-vitro susceptibility testing of Streptococccus pneumoniae. This study attempts to determine the minimum inhibitory concentration (MIC of second-generation (ciprofloxacin and ofloxacin, third-generation (levofloxacin and the fourth-generation (moxifloxacin and gatifloxacin fluoroquinolones against S. pneumoniae recovered from bacterial keratitis. Materials and Methods: In retrospect, the MICs of 50 strains of S. pneumoniae isolated from the corneal scrapes of patients with bacterial keratitis were determined against ciprofloxacin, ofloxacin, levofloxacin, gatifloxacin and moxifloxacin using E-tests. The National Committee of Clinical Laboratory Standards (NCCLS susceptibility patterns and the potencies of the MICs were statistically compared. Results: The median MIC of ciprofloxacin (0.25μg/ml was found to be lower than the median MICs of ofloxacin (0.5μg/ml (P < 0.449 and levofloxacin (1.0μg/ml (P < 0.001. The median MICs of gatifloxacin (0.1μg/ml was lower than the median MICs of ciprofloxacin (0.25μg/ml (P < 0.001, ofloxacin (0.5μg/ml (P < 0.001 and levofloxacin (1.0μg/ml (P < 0.001. Moxifloxacin (0.06μg/ml had showed lower median MICs than gatifloxacin (0.1μg/ml (P < 0.001 levofloxacin (1.0μg/ml (P < 0.001, ofloxacin (0.5μg/ml (P < 0.001 and ciprofloxacin (0.25μg/ml (P < 0.001. Moxifloxacin (0.06μg/ml had a lower MIC50 (μg/ml than gatifloxacin (0.1μg/ml, levofloxacin (1.0μg/ml, ciprofloxacin (0.25μg/ml and ofloxacin (0.5μg/ml. MIC90 (μg/ml of moxifloxacin (0.06μg/ml was found to be lower than the MIC90 (μg/ml of gatifloxacin (0.5μg/ml, levofloxacin (1.0μg/ml, ofloxacin (0.5μg/ml and ciprofloxacin (0.5μg/ml. Conclusion: Based on in-vitro testing, the five portrayed fluoroquinolones 100% sensitivity to S. pneumoniae. However, the fourth-generation fluoroquinolone, moxifloxacin appeared to be more

  13. The impact of pneumococcal conjugate vaccines on carriage of and disease caused by Streptococcus pneumoniae serotypes 6C and 6D in southern Israel.

    Porat, Nurith; Benisty, Rachel; Givon-Lavi, Noga; Trefler, Ronit; Dagan, Ron

    2016-05-27

    The introduction of the 7-valent pneumococcal conjugate vaccine (PCV7) followed by PCV13 resulted in a dramatic reduction in carriage and disease rates of Streptococcus pneumoniae (Sp) serotype 6B (Sp6B) and Sp6A. The structural modifications of the capsule of Sp6A and Sp6B to become Sp6C and Sp6D, respectively, raised a concern that eradication of Sp6A/Sp6B by PCV could be accompanied by an increase in Sp6C/Sp6D. This study examines the dynamics and clonal distribution of Sp6C/Sp6D relative to Sp6A/Sp6B during 1999-2014, pre- and post-PCV implementation. Sp were cultured from Blood/CSF and MEF of children pneumococcal disease, complete elimination of serogroup 6 was found in the PCV era. Similar clonal composition was found for Sp6C and Sp6D pre- and post-PCV. We conclude that Sp6C and Sp6D do not act as replacement serotypes for Sp6A and Sp6B following vaccination with PCV13. The major Sp6C and Sp6D clones present pre-PCV persisted also post-PCV implementation, suggesting that these clones possess an advantage retained post-vaccination. PMID:27113163

  14. Invasive Streptococcus pneumoniae in Canada, 2011-2014: Characterization of new candidate 15-valent pneumococcal conjugate vaccine serotypes 22F and 33F.

    Golden, Alyssa R; Adam, Heather J; Zhanel, George G

    2016-05-17

    Emerging non-PCV-13 Streptococcus pneumoniae serotypes 22F and 33F are included in a new 15-valent pneumococcal conjugate vaccine currently undergoing clinical trials in the United States. This study assessed the antimicrobial resistance and genetic relatedness of these two emerging pneumococcal serotypes. Of the 5075 invasive pneumococcal isolates collected in Canada from 2011 to 2014, 9.8% (497/5075) were serotype 22F and 3.2% (160/5075) were serotype 33F. Despite being among the top 4 most common serotypes collected each study year, serotype 22F demonstrated ≥98% susceptibility to all antimicrobials tested except clarithromycin and few were multi-drug resistant (MDR) (0.8%, 4/497). Serotype 22F isolates were highly clonal (ST433), with two isolates showing high relatedness to MDR international clone Sweden(15A)-25 (ST63). Conversely, serotype 33F showed greater antimicrobial resistance, greater genetic diversity and a higher proportion of MDR isolates (8.8%, 14/160). The prevalence of serotype 33F increased significantly during 2011-2014 (p=0.005). PMID:27085174

  15. Acyl-Acyl carrier protein regulates transcription of fatty acid biosynthetic genes via the FabT repressor in Streptococcus pneumoniae.

    Jerga, Agoston; Rock, Charles O

    2009-06-01

    Long-chain acyl-acyl carrier proteins (acyl-ACP) are established biochemical regulators of bacterial type II fatty acid synthases due to their ability to feedback-inhibit the early steps in the biosynthetic pathway. In Streptococcus pneumoniae, the expression of the fatty acid synthase (fab) genes is controlled by a helix-turn-helix transcriptional repressor called FabT. A screen of pathway intermediates identified acyl-ACP as a ligand that increased the affinity of FabT for DNA. FabT bound to a wide range of acyl-ACP chain lengths in the absence of DNA, but only the long-chain acyl-ACPs increase the affinity of FabT for DNA. FabT affinity for DNA increased with increasing acyl-ACP chain length with cis-vaccenoyl-ACP being the most effective ligand. Thus, FabT is a new ACP-interacting partner that acts as a transcriptional rheostat to fine tune the expression of the fab genes based on the demand for fatty acids. PMID:19376778

  16. Acyl-Acyl Carrier Protein Regulates Transcription of Fatty Acid Biosynthetic Genes via the FabT Repressor in Streptococcus pneumoniae*

    Jerga, Agoston; Rock, Charles O.

    2009-01-01

    Long-chain acyl-acyl carrier proteins (acyl-ACP) are established biochemical regulators of bacterial type II fatty acid synthases due to their ability to feedback-inhibit the early steps in the biosynthetic pathway. In Streptococcus pneumoniae, the expression of the fatty acid synthase (fab) genes is controlled by a helix-turn-helix transcriptional repressor called FabT. A screen of pathway intermediates identified acyl-ACP as a ligand that increased the affinity of FabT for DNA. FabT bound to a wide range of acyl-ACP chain lengths in the absence of DNA, but only the long-chain acyl-ACPs increase the affinity of FabT for DNA. FabT affinity for DNA increased with increasing acyl-ACP chain length with cis-vaccenoyl-ACP being the most effective ligand. Thus, FabT is a new ACP-interacting partner that acts as a transcriptional rheostat to fine tune the expression of the fab genes based on the demand for fatty acids. PMID:19376778

  17. 肺炎链球菌毒力因子研究新进展%Research progress of virulence factors of Streptococcus pneumoniae

    刘丽; 刘兆明; 朱德全; 冯尚彩

    2013-01-01

    Streptococcus pneumonia (Spn) is the major pathogen of human disease.There are several virulence factors that are involved in the pathogenicity of Spn.With genetic studies and mutant strain applications,it contributes to a deep understanding of the classic pneunmococcal virulence factors and identification of novel virulence factors.This review discusses the biological activity and pathogenic mechanism of several pneumococcal virulence factors,including polysaccharide capsule,pilus,proteins and proteases.%肺炎链球菌(Spn)是临床重要的致病菌.Spn致病性与很多毒力因子有关.随着基因组学的研究和变异菌株的应用,人们除了更深入地了解Spn经典毒力因子外,也认识了其新的毒力因子.此文针对Spn毒力因子,包括荚膜多糖、菌毛、蛋白和蛋白酶等,对其生物活性和在菌株致病过程中的作用机制进行综述.

  18. Reduced turnaround time and improved diagnosis of invasive serogroup B Neisseria meningitidis and Streptococcus pneumoniae infections using a lyophilized quadruplex quantitative PCR.

    McHugh, Martin P; Gray, Steve J; Kaczmarski, Edward B; Guiver, Malcolm

    2015-11-01

    Since 1996 the Meningococcal Reference Unit (MRU) in Manchester has provided a national service for PCR confirmation of meningococcal and pneumococcal disease. Neisseria meningitidis serogroup B is predominant in the UK, accounting for >60% of cases. In response to this, the MRU has developed a quadruplex quantitative PCR that detects N. meningitidis capsule transporter (ctrA), serogroup B sialyltransferase (siaDB), Streptococcus pneumoniae pneumolysin (ply) and an internal control. The assay was prepared in a ready-to-use lyophilized format by Applied Biosystems. Laboratory validation showed excellent performance in a specificity panel of 52 isolates and improved detection in comparison with the routine assay. Testing of 244 patient samples showed sensitivity of 93% [95% confidence interval (CI): 88-98%] for the ctrA assay, 95% (95% CI: 91-100%) for the siaDB assay and 100% (95% CI: 95-100%) for the ply assay. Specificity was 100% (95% CI: 98-100%) for both meningococcal targets and 95% (95% CI: 92-98%) for ply. The quadruplex also retained high performance in mixed samples and had acceptable reproducibility. After introduction of the quadruplex into routine use the turnaround time for N. meningitidis group B PCR confirmation reduced from 37 to 29 h and the internal control has proved useful for detecting inhibitory samples. The quadruplex assay provides rapid group B confirmation of meningococcal positive samples, enabling timely public health interventions for the most common disease-causing meningococcal serogroup in the UK. PMID:26253287

  19. A novel high-throughput method for molecular serotyping and serotype-specific quantification of Streptococcus pneumoniae using a nanofluidic real-time PCR system.

    Dhoubhadel, Bhim Gopal; Yasunami, Michio; Yoshida, Lay-Myint; Thi, Hien Anh Nguyen; Thi, Thu Huong Vu; Thi, Thuy Ai Nguyen; Watanabe, Kiwao; Suzuki, Motoi; Morimoto, Konosuke; Dang, Duc Anh; Ariyoshi, Koya

    2014-04-01

    Serotype-specific quantification data are essential for elucidating the complex epidemiology of Streptococcus pneumoniae and evaluating pneumococcal vaccine efficacy. Various PCR-based assays have been developed to circumvent the drawback of labour-intensive and time-consuming culture-based procedures for serotype determination and quantification of pneumococcus. Here, we applied a nanofluidic real-time PCR system to establish a novel assay. Twenty-nine primer pairs, 13 of which were newly designed, were selected for the assay to cover 50 serotypes including all currently available conjugate and polysaccharide vaccine serotypes. All primer pairs were evaluated for their sensitivity, specificity, efficiency, repeatability, accuracy and reproducibility on the Fluidigm Biomark HD System, a nanofluidic real-time PCR system, by drawing standard curves with a serial dilution of purified DNA. We applied the assay to 52 nasopharyngeal swab samples from patients with pneumonia confirmed by chest X-ray to validate its accuracy. Minimum detection levels of this novel assay using the nanofluidic real-time PCR system were comparable to the conventional PCR-based assays (between 30 and 300 copies per reaction). They were specific to their targets with good repeatability (sd of copy number of 0.1), accuracy (within ±0.1 fold difference in log10 copy number) and reproducibility (sd of copy number of 0.1). When artificially mixed DNA samples consisting of multiple serotypes in various ratios were tested, all the serotypes were detected proportionally, including a minor serotype of one in 1000 copies. In the nasopharyngeal samples, the PCR system detected all the culture-positive samples and 22 out of 23 serotypes identified by the conventional method were matched with PCR results. We conclude that this novel assay, which is able to differentially quantify 29 pneumococcus groups for 45 test samples in a single run, is applicable to the large-scale epidemiological study of

  20. Sorotipos de Streptococcus pneumoniae isolados de líquido cefalorraquidiano no período de 1977-1988 na cidade de São Paulo, Brasil Serotypes of Streptococcus pneumoniae isolated from cerebrospinal fluid

    A.E. Taunay

    1990-02-01

    Full Text Available Desde 1977, o Instituto Adolfo Lutz (IAL vem promovendo a sorotipagem do S. pneumoniae ou pneumococo de infecções causadas por esta bacteria. As cepas isoladas têm sido encaminhadas ao WHO Pneumococcal Reference Center, Pensilvania, E.U.A.. De 1977 a 1988, 1.000 cepas de pneumococo isoladas de LCR foram sorotipadas, de acordo com a nomenclatura dinamarquesa, e 60 sorotipos foram identificados. A maior freqüência foi do sorotipo 1, secundado por 6B, 18C, 14, 5, 3, 6A, 23F, 19F e 38. Estes sorotipos distribuídos segundo faixas etárias demonstraram incidência variável, notando-se uma certa peculiaridade, ou seja, a predominância do sorotipo 6B na faixa de zero a menos de dois anos; do sorotipo 1 na faixa de 2 até 50 anos e do sorotipo 3 no grupo acima de 50 anos. Nos 12 anos considerados, 25 sorotipos apresentaram uma certa uniformidade na freqüência e o mesmo foi observado com relação às estações climáticas, apenas com um número maior de infecções meníngeas nos meses mais frios. Considerando a gravidade das infecções pneumocócicas notadamente as meningites, e a pouca informação relativa aos sorotipos pneumocócicos que ocorrem na região, julgamos importante essa informação relativa aos sorotipos, uma vez que tem sido usadas, com sucesso, vacinas polissacarídicas na prevenção dessas infecções.Since 1977, the Instituto Adolfo Lutz (IAL is having interest in the scrotyping of S. pneumoniae or pneumococcus from infections caused by this bacteria. The isolated strains have been sent to the WHO Pneumococcal Reference Center, Pennsylvania, U.S.A.. From 1977 to 1988, 1.000 pneumococcus strains isolated from cerebrospinal fluid were typed, according to Danish nomenclature, and 60 serotypes were identified. The most frequent serotypes were 1, 6B, 18C, 14, 5, 3, 6A, 23F, 19F, and 38. Among different age groups, they showed a variable incidence, with serotype 6B in the ages of zero to almost 2 years old, serotype 1 in the

  1. Streptococcus pneumoniae: vigilancia molecular de aislamientos invasivos resistentes a penicilina recuperados de julio 2003 a junio 2004 en Colombia

    Moreno Jaime

    2005-07-01

    Full Text Available Las infecciones neumocócicas afectan principalmente a niños y ancianos. Además, por el incremento de aislamientos resistentes a penicilina, S. pneumoniae es considerado como uno de los principales problemas de salud pública. El objetivo fue determinar las relaciones genéticas de los aislamientos
    invasores de S. pneumoniae con susceptibilidad disminuida a penicilina (SDP recuperados de julio-2003 a junio-2004. Se estudiaron 66 aislamientos utilizando la técnica de electroforesis de campo pulsado (PFGE, los patrones electroforéticos se compararon según los criterios de
    Tenover y se analizaron con el programa Fingerprinting TMII 3.0. En 12 aislamientos se identificaron los perfiles PBP, con la técnica del polimorfismo en la longitud de los fragmentos de restricción (RFLP de los genes pbp 2b, 2x y 1a, los cuales se interpretaron según lo establecido por el laboratorio. Las relaciones entre los grupos clonales y las características demográficas de los pacientes se analizaron con los programas EpiInfo 6.04 y MVSP 3.1. La mayoría de los aislamientos (71%
    presentaron el patrón PFGE B relacionado con el clon 3-España9V, seguido por los patrones C (6% agrupado con el clon 26-Colombia23F, D (4% con el clon 2-España6B y A (2% con el clon 1-España23F, los aislamientos restantes se distribuyeron en diez patrones no relacionados con clones internacionales. En los aislamientos relacionados con los clones 1, 2, y 26 se identificó el mismo perfil PBP del clon y los relacionados con el clon 3 presentaron una variante del perfil
    PBP del clon 3. En Colombia la prevalencia de aislamientos invasores resistentes a penicilina se debe a la circulación de los clones internacionales 1-España23F, 2-España6B, 3-España9V y
    26-Colombia23F.

  2. Streptococcus pneumoniae Serotypes and Mortality in Adults and Adolescents in South Africa: Analysis of National Surveillance Data, 2003 - 2008.

    Cheryl Cohen

    Full Text Available An association between pneumococcal serotypes and mortality has been suggested. We aimed to investigate this among individuals aged ≥15 years with invasive pneumococcal disease (IPD in South Africa.IPD cases were identified through national laboratory-based surveillance at 25 sites, pre-pneumococcal conjugate vaccine (PCV introduction, from 2003-2008. We assessed the association between the 20 commonest serotypes and in-hospital mortality using logistic regression with serotype 4 (the third commonest serotype with intermediate case-fatality ratio (CFR as referent.Among 3953 IPD cases, CFR was 55% (641/1166 for meningitis and 23% (576/2484 for bacteremia (p<0.001. Serotype 19F had the highest CFR (48%, 100/207, followed by serotype 23F (39%, 99/252 and serotype 1 (38%, 246/651. On multivariable analysis, factors independently associated with mortality included serotype 1 (OR 1.9, 95%CI 1.1-3.5 and 19F (OR 2.9, 95%CI 1.4-6.1 vs. serotype 4; increasing age (25-44 years, OR 1.8, 95%CI 1.0-3.0; 45-64 years, OR 3.6, 95%CI 2.0-6.4; ≥65 years, OR 5.2, 95%CI 1.9-14.1; vs. 15-24 years; meningitis (OR 4.1, 95%CI 3.0-5.6 vs. bacteremic pneumonia; and HIV infection (OR1.7, 95%CI 1.0-2.8. On stratified multivariate analysis, serotype 19F was associated with increased mortality amongst bacteremic pneumococcal pneumonia cases, while no serotype was associated with increased mortality in meningitis cases.Mortality was increased in HIV-infected individuals, which may be reduced by increased antiretroviral therapy availability. Serotypes associated with increased mortality are included in the 10-and-13-valent PCV and may become less common in adults due to indirect effects following routine infant immunization.

  3. Síndrome hemolítico-urêmica relacionada à infecção invasiva pelo Streptococcus pneumoniae Hemolytic-uremic syndrome complicating invasive pneumococcal disease

    Anna Leticia de O. Cestari

    2008-03-01

    Full Text Available OBJETIVO: A doença pneumocócica é importante problema de saúde pública e raramente há associação desta infecção com a síndrome hemolítico-urêmica (SHU grave. O objetivo deste artigo é relatar o caso de um paciente com esta associação. DESCRIÇÃO DO CASO: Criança do sexo masculino, com 17 meses de idade, admitida no hospital com insuficiência respiratória aguda e necessitando de suporte ventilatório. O exame radiológico mostrava extensa opacidade homogênea em hemitórax direito. A hemocultura foi positiva para Streptococcus pneumoniae. Nos exames de admissão, notaram-se: hemoglobina de 6,5g/dL, 38.000 plaquetas/mm³, uréia de 79mg/dL e creatinina de 1,64mg/dL. No primeiro dia, apresentou oligoanúria e hipervolemia, necessitando de hemodiafiltração. Evoluiu com disfunção de múltiplos órgãos e óbito no sétimo dia. A necrópsia mostrou áreas extensas de necrose cortical e tubular renal, com depósito de fibrina nas arteríolas. COMENTÁRIOS: A SHU associada ao pneumococo apresenta morbidade e mortalidade elevadas. Em crianças com doença pneumocócica invasiva e acometimento hematológico ou renal grave, deve-se estar atento a esta rara complicação. Merecem investigação os seguintes aspectos relacionados à doença: a função da detecção precoce de antígenos T ativados no diagnóstico e terapêutica, o papel do fator H na patogênese, o método ideal de substituição renal e a definição do prognóstico em longo prazo.OBJECTIVE: Pneumococcal diseases are a major public health problem. Severe hemolytic-uremic syndrome is an uncommon complication. The aim of this study is to report a child with this complication. CASE DESCRIPTION: A male child with 17 months old was admitted to the hospital, due to acute respiratory failure, needing ventilatory support. Roentgenogram demonstrated massive condensation of right lung and Streptococcus pneumonia was isolated from blood cultures. Laboratory tests showed

  4. Crystal structure of enoyl-acyl carrier protein reductase (FabK) from Streptococcus pneumoniae reveals the binding mode of an inhibitor.

    Saito, Jun; Yamada, Mototsugu; Watanabe, Takashi; Iida, Maiko; Kitagawa, Hideo; Takahata, Sho; Ozawa, Tomohiro; Takeuchi, Yasuo; Ohsawa, Fukuichi

    2008-04-01

    Enoyl-acyl carrier protein (ACP) reductases are critical for bacterial type II fatty acid biosynthesis and thus are attractive targets for developing novel antibiotics. We determined the crystal structure of enoyl-ACP reductase (FabK) from Streptococcus pneumoniae at 1.7 A resolution. There was one dimer per asymmetric unit. Each subunit formed a triose phosphate isomerase (TIM) barrel structure, and flavin mononucleotide (FMN) was bound as a cofactor in the active site. The overall structure was similar to the enoyl-ACP reductase (ER) of fungal fatty acid synthase and to 2-nitropropane dioxygenase (2-ND) from Pseudomonas aeruginosa, although there were some differences among these structures. We determined the crystal structure of FabK in complex with a phenylimidazole derivative inhibitor to envision the binding site interactions. The crystal structure reveals that the inhibitor binds to a hydrophobic pocket in the active site of FabK, and this is accompanied by induced-fit movements of two loop regions. The thiazole ring and part of the ureido moiety of the inhibitor are involved in a face-to-face pi-pi stacking interaction with the isoalloxazine ring of FMN. The side-chain conformation of the proposed catalytic residue, His144, changes upon complex formation. Lineweaver-Burk plots indicate that the inhibitor binds competitively with respect to NADH, and uncompetitively with respect to crotonoyl coenzyme A. We propose that the primary basis of the inhibitory activity is competition with NADH for binding to FabK, which is the first step of the two-step ping-pong catalytic mechanism. PMID:18305197

  5. Two distinct functions of ComW in stabilization and activation of the alternative sigma factor ComX in Streptococcus pneumoniae.

    Sung, Chang Kyoo; Morrison, Donald A

    2005-05-01

    Natural genetic transformation in Streptococcus pneumoniae is controlled by a quorum-sensing system, which acts through the competence-stimulating peptide (CSP) for transient activation of genes required for competence. More than 100 genes have been identified as CSP regulated by use of DNA microarray analysis. One of the CSP-induced genes required for genetic competence is comW. As the expression of this gene depended on the regulator ComE, but not on the competence sigma factor ComX (sigma(X)), and as expression of several genes required for DNA processing was affected in a comW mutant, comW appears to be a new regulatory gene. Immunoblotting analysis showed that the amount of the sigma(X) protein is dependent on ComW, suggesting that ComW may be directly or indirectly involved in the accumulation of sigma(X). As sigma(X) is stabilized in clpP mutants, a comW mutation was introduced into the clpP background to ask whether the synthesis of sigma(X) depends on ComW. The clpP comW double mutant accumulated an amount of sigma(X) higher (threefold) than that seen in the wild type but was not transformable, suggesting that while comW is not needed for sigma(X) synthesis, it acts both in stabilization of sigma(X) and in its activation. Modification of ComW with a histidine tag at its C or N terminus revealed that both amino and carboxyl termini are important for increasing the stability of sigma(X), but only the N terminus is important for stimulating its activity. PMID:15838032

  6. Suppression of a deletion mutation in the gene encoding essential PBP2b reveals a new lytic transglycosylase involved in peripheral peptidoglycan synthesis in Streptococcus pneumoniae D39.

    Tsui, Ho-Ching Tiffany; Zheng, Jiaqi J; Magallon, Ariel N; Ryan, John D; Yunck, Rachel; Rued, Britta E; Bernhardt, Thomas G; Winkler, Malcolm E

    2016-06-01

    In ellipsoid-shaped ovococcus bacteria, such as the pathogen Streptococcus pneumoniae (pneumococcus), side-wall (peripheral) peptidoglycan (PG) synthesis emanates from midcells and is catalyzed by the essential class B penicillin-binding protein PBP2b transpeptidase (TP). We report that mutations that inactivate the pneumococcal YceG-domain protein, Spd_1346 (renamed MltG), remove the requirement for PBP2b. ΔmltG mutants in unencapsulated strains accumulate inactivation mutations of class A PBP1a, which possesses TP and transglycosylase (TG) activities. The 'synthetic viable' genetic relationship between Δpbp1a and ΔmltG mutations extends to essential ΔmreCD and ΔrodZ mutations that misregulate peripheral PG synthesis. Remarkably, the single MltG(Y488D) change suppresses the requirement for PBP2b, MreCD, RodZ and RodA. Structural modeling and comparisons, catalytic-site changes and an interspecies chimera indicate that pneumococcal MltG is the functional homologue of the recently reported MltG endo-lytic transglycosylase of Escherichia coli. Depletion of pneumococcal MltG or mltG(Y488D) increases sphericity of cells, and MltG localizes with peripheral PG synthesis proteins during division. Finally, growth of Δpbp1a ΔmltG or mltG(Y488D) mutants depends on induction of expression of the WalRK TCS regulon of PG hydrolases. These results fit a model in which MltG releases anchored PG glycan strands synthesized by PBP1a for crosslinking by a PBP2b:RodA complex in peripheral PG synthesis. PMID:26933838

  7. The relBE2Spn toxin-antitoxin system of Streptococcus pneumoniae: role in antibiotic tolerance and functional conservation in clinical isolates.

    Concha Nieto

    Full Text Available Type II (proteic chromosomal toxin-antitoxin systems (TAS are widespread in Bacteria and Archaea but their precise function is known only for a limited number of them. Out of the many TAS described, the relBE family is one of the most abundant, being present in the three first sequenced strains of Streptococcus pneumoniae (D39, TIGR4 and R6. To address the function of the pneumococcal relBE2Spn TAS in the bacterial physiology, we have compared the response of the R6-relBE2Spn wild type strain with that of an isogenic derivative, Delta relB2Spn under different stress conditions such as carbon and amino acid starvation and antibiotic exposure. Differences on viability between the wild type and mutant strains were found only when treatment directly impaired protein synthesis. As a criterion for the permanence of this locus in a variety of clinical strains, we checked whether the relBE2Spn locus was conserved in around 100 pneumococcal strains, including clinical isolates and strains with known genomes. All strains, although having various types of polymorphisms at the vicinity of the TA region, contained a functional relBE2Spn locus and the type of its structure correlated with the multilocus sequence type. Functionality of this TAS was maintained even in cases where severe rearrangements around the relBE2Spn region were found. We conclude that even though the relBE2Spn TAS is not essential for pneumococcus, it may provide additional advantages to the bacteria for colonization and/or infection.

  8. High-Resolution Crystal Structures of Streptococcus pneumoniae Nicotinamidase with Trapped Intermediates Provide Insights into the Catalytic Mechanism and Inhibition by Aldehydes

    French, Jarrod B.; Cen, Yana; Sauve, Anthony A.; Ealick, Steven E. (Cornell); (Weill-Med)

    2010-11-11

    Nicotinamidases are salvage enzymes that convert nicotinamide to nicotinic acid. These enzymes are essential for the recycling of nicotinamide into NAD{sup +} in most prokaryotes and most single-cell and multicellular eukaryotes, but not in mammals. The significance of these enzymes for nicotinamide salvage and for NAD{sup +} homeostasis has stimulated interest in nicotinamidases as possible antibiotic targets. Nicotinamidases are also regulators of intracellular nicotinamide concentrations, thereby regulating signaling of downstream NAD{sup +}-consuming enzymes, such as the NAD{sup +}-dependent deacetylases (sirtuins). Here, we report several high-resolution crystal structures of the nicotinamidase from Streptococcus pneumoniae (SpNic) in unliganded and ligand-bound forms. The structure of the C136S mutant in complex with nicotinamide provides details about substrate binding, while a trapped nicotinoyl thioester in a complex with SpNic reveals the structure of the proposed thioester reaction intermediate. Examination of the active site of SpNic reveals several important features, including a metal ion that coordinates the substrate and the catalytically relevant water molecule and an oxyanion hole that both orients the substrate and offsets the negative charge that builds up during catalysis. Structures of this enzyme with bound nicotinaldehyde inhibitors elucidate the mechanism of inhibition and provide further details about the catalytic mechanism. In addition, we provide a biochemical analysis of the identity and role of the metal ion that orients the ligand in the active site and activates the water molecule responsible for hydrolysis of the substrate. These data provide structural evidence of several proposed reaction intermediates and allow for a more complete understanding of the catalytic mechanism of this enzyme.

  9. Physical structure and genetic expression of the sulfonamide-resistance plasmid pLS80 and its derivatives in Streptococcus pneumoniae and Bacillus subtilis

    Lopez, P.; Espinosa, M.; Lacks, S.A.

    1984-01-01

    The 10-kb chromosomal fragment of Streptococcus pneumoniae cloned in pLS80 contains the sul-d allele of the pneumococcal gene for dihydropteroate synthase. As a single copy in the chromosome this allele confers resistance to sulfanilamide at 0.2 mg/ml; in the multicopy plasmid it confers resistance to 2.0 mg/ml. The sul-d mutation was mapped by restriction analysis to a 0.4-kb region. A spontaneous deletion beginning approx. 1.5 kb to the right of the sul-d mutation prevented gene function, possibly by removing a promoter. This region could be restored by chromosomal facilitation and be demonstrated in the plasmid by selection for sulfonamide resistance. Under selection for a vector marker, tetracycline resistance, only the deleted plasmid was detectable, apparently as a result of plasmid segregation and the advantageous growth rates of cells with smaller plasmids. When such cells were selected for sulfonamide resistance, the deleted region returned to the plasmid, presumably by equilibration between the chromosome and the plasmid pool, to give a low frequency (approx. 10/sup -3/) of cells resistant to sulfanilamide at 2.0 mg/ml. Models for the mechanisms of chromosomal facilitation and equilibration are proposed. Several derivatives of pLS80 could be transferred to Bacillus subtilis, where they conferred resistance to sulfanilamide at 2 mg/ml, thereby demonstrating cross-species expression of the pneumococcal gene. Transfer of the plasmids to B. subtilis gave rise to large deletions to the left of the sul-d marker, but these deletions did not interfere with the sul-d gene function. Restriction maps of pLS80 and its variously deleted derivatives are presented.

  10. Familias de la proteína de superficie PspA de Streptococcus pneumoniae: Relación con serotipos y localización

    Clara Mayoral

    2010-10-01

    Full Text Available PspA, proteína de superficie de Streptococcus pneumoniae es un factor de virulencia, fuertemente inmunogénica y común a todos los serotipos. Aunque el gen que codifica para esta proteína presenta una marcada heterogeneidad en la región correspondiente al N-terminal, la PspA contiene epitopes conservados de manera tal que la inmunización genera protección contra neumococos pertenecientes a diversos tipos capsulares y con distintas PspA. A pesar del marcado polimorfismo del gen pspA es posible agrupar las distintas variantes en 3 familias mayoritarias. Estas propiedades las convierten en candidatas ideales para elaborar vacunas. Debido a que la mayoría de los trabajos sobre identificación de familias fueron realizados sobre serotipos frecuentes en otros países, el objetivo fue identificar las familias de PspA de aislamientos de pacientes de nuestra región y relacionarlas con los serotipos prevalentes y patologías. Se estudiaron 70 aislamientos, provenientes de niños con infecciones invasoras. Se aplicó una PCR utilizando cebadores específicos de cada familia. El 60% fueron familia 1 y 34% familia 2. En un 6% no se identificó ninguna de las familias de PspA. Los serotipos 1 y 5 presentaron familia 1 únicamente; los serotipos 14, 6B, 19F y 18C mostraron genes de ambas familias. La familia 1 se observó en 60% de las neumonías y 50% de las meningitis. La familia 2 en 33% de neumonías y 50% de meningitis. Esta información podría ser un valioso aporte para la formulación de una vacuna regional efectiva utilizando PspA recombinante como inmunógeno.

  11. Crystallization and preliminary X-ray diffraction studies of choline-binding protein F from Streptococcus pneumoniae

    Molina, Rafael [Grupo de Cristalografía Macromolecular y Biología Estructural, Instituto Química Física Rocasolano, CSIC, Serrano 119, 28006 Madrid (Spain); González, Ana; Moscoso, Miriam; García, Pedro [Departamento de Microbiología Molecular, Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, 28040 Madrid (Spain); Stelter, Meike; Kahn, Richard [Institut de Biologie Structurale J.-P. Ebel CEA CNRS UJF, Laboratoire de Cristallographie Macromoléculaire, 41 Rue Jules Horowitz, 38027 Grenoble CEDEX 1 (France); Hermoso, Juan A., E-mail: xjuan@iqfr.csic.es [Grupo de Cristalografía Macromolecular y Biología Estructural, Instituto Química Física Rocasolano, CSIC, Serrano 119, 28006 Madrid (Spain)

    2007-09-01

    The modular choline-binding protein F (CbpF) from S. pneumoniae has been crystallized by the hanging-drop vapour-diffusion method. A SAD data set from a gadolinium-complex derivative has been collected to 2.1 Å resolution. Choline-binding protein F (CbpF) is a modular protein that is bound to the pneumococcal cell wall through noncovalent interactions with choline moieties of the bacterial teichoic and lipoteichoic acids. Despite being one of the more abundant proteins on the surface, along with the murein hydrolases LytA, LytB, LytC and Pce, its function is still unknown. CbpF has been crystallized using the hanging-drop vapour-diffusion method at 291 K. Diffraction-quality orthorhombic crystals belong to space group P2{sub 1}2{sub 1}2, with unit-cell parameters a = 49.13, b = 114.94, c = 75.69 Å. A SAD data set from a Gd-HPDO3A-derivatized CbpF crystal was collected to 2.1 Å resolution at the gadolinium L{sub III} absorption edge using synchrotron radiation.

  12. Diverse Virulent Pneumophages Infect Streptococcus mitis

    Siham Ouennane; Philippe Leprohon; Sylvain Moineau

    2015-01-01

    Streptococcus mitis has emerged as one of the leading causes of bacterial endocarditis and is related to Streptococcus pneumoniae. Antibiotic resistance has also increased among strains of S. mitis and S. pneumoniae. Phages are being reinvestigated as alternatives to antibiotics for managing infections. In this study, the two virulent phages Cp-1 (Podoviridae) and Dp-1 (Siphoviridae), previously isolated from S. pneumoniae, were found to also infect S. mitis. Microbiological assays showed tha...

  13. Population genetic structure of Streptococcus pneumoniae in Kilifi, Kenya, prior to the introduction of pneumococcal conjugate vaccine.

    Angela B Brueggemann

    Full Text Available The 10-valent pneumococcal conjugate vaccine (PCV10 was introduced in Kenya in 2011. Introduction of any PCV will perturb the existing pneumococcal population structure, thus the aim was to genotype pneumococci collected in Kilifi before PCV10.Using multilocus sequence typing (MLST, we genotyped >1100 invasive and carriage pneumococci from children, the largest collection genotyped from a single resource-poor country and reported to date. Serotype 1 was the most common serotype causing invasive disease and was rarely detected in carriage; all serotype 1 isolates were members of clonal complex (CC 217. There were temporal fluctuations in the major circulating sequence types (STs; and although 1-3 major serotype 1, 14 or 23F STs co-circulated annually, the two major serotype 5 STs mainly circulated independently. Major STs/CCs also included isolates of serotypes 3, 12F, 18C and 19A and each shared ≤ 2 MLST alleles with STs that circulate widely elsewhere. Major CCs associated with non-PCV10 serotypes were predominantly represented by carriage isolates, although serotype 19A and 12F CCs were largely invasive and a serotype 10A CC was equally represented by invasive and carriage isolates.Understanding the pre-PCV10 population genetic structure in Kilifi will allow for the detection of changes in prevalence of the circulating genotypes and evidence for capsular switching post-vaccine implementation.

  14. Characterization of 19A-like 19F pneumococcal isolates from Papua New Guinea and Fiji.

    Dunne, E M; Tikkanen, L; Balloch, A; Gould, K; Yoannes, M; Phuanukoonnon, S; Licciardi, P V; Russell, F M; Mulholland, E K; Satzke, C; Hinds, J

    2015-09-01

    Molecular identification of Streptococcus pneumoniae serotype 19F is routinely performed by PCR targeting the wzy gene of the capsular biosynthetic locus. However, 19F isolates with genetic similarity to 19A have been reported in the United States and Brazil. We screened 78 pneumococcal carriage isolates and found six 19F wzy variants that originated from children in Papua New Guinea and Fiji. Isolates were characterized using multilocus sequence typing and opsonophagocytic assays. The 19F wzy variants displayed similar susceptibility to anti-19F IgG antibodies compared to standard 19F isolates. Our findings indicate that these 19F variants may be more common than previously believed. PMID:26339490

  15. LacR Is a Repressor of lacABCD and LacT Is an Activator of lacTFEG, Constituting the lac Gene Cluster in Streptococcus pneumoniae

    Afzal, Muhammad; Shafeeq, Sulman; Kuipers, Oscar P

    2014-01-01

    Comparison of the transcriptome of Streptococcus pneumoniae strain D39 grown in the presence of either lactose or galactose with that of the strain grown in the presence of glucose revealed the elevated expression of various genes and operons, including the lac gene cluster, which is organized into two operons, i.e., lac operon I (lacABCD) and lac operon II (lacTFEG). Deletion of the DeoR family transcriptional regulator lacR that is present downstream of the lac gene cluster revealed elevate...

  16. PspA family distribution, antimicrobial resistance and serotype of Streptococcus pneumoniae isolated from upper respiratory tract infections in Japan.

    Muneki Hotomi

    Full Text Available BACKGROUND: The protection against pneumococcal infections provided by currently available pneumococcal polysaccharide conjugate vaccines are restricted to the limited number of the serotypes included in the vaccine. In the present study, we evaluated the distribution of the pneumococcal capsular type and surface protein A (PspA family of pneumococcal isolates from upper respiratory tract infections in Japan. METHODS: A total of 251 S. pneumoniae isolates from patients seeking treatment for upper respiratory tract infections were characterized for PspA family, antibiotic resistance and capsular type. RESULTS: Among the 251 pneumococci studied, the majority (49.4% was identified as belonging to PspA family 2, while most of the remaining isolates (44.6% belonged to family 1. There were no significant differences between the distributions of PspA1 versus PspA2 isolates based on the age or gender of the patient, source of the isolates or the isolates' susceptibilities to penicillin G. In contrast, the frequency of the mefA gene presence and of serotypes 15B and 19F were statistically more common among PspA2 strains. CONCLUSION: The vast majority of pneumococci isolated from the middle ear fluids, nasal discharges/sinus aspirates or pharyngeal secretions represented PspA families 1 and 2. Capsular serotypes were generally not exclusively associated with certain PspA families, although some capsular types showed a much higher proportion of either PspA1 or PspA2. A PspA-containing vaccine would potentially provide high coverage against pneumococcal infectious diseases because it would be cross-protective versus invasive disease with the majority of pneumococci infecting children and adults.

  17. Serotypes and Clonal Diversity of Streptococcus pneumoniae Causing Invasive Disease in the Era of PCV13 in Catalonia, Spain.

    Eva Del Amo

    Full Text Available The aim of this study was to study the serotypes and clonal diversity of pneumococci causing invasive pneumococcal disease in Catalonia, Spain, in the era of 13-valent pneumococcal conjugate vaccine (PCV13. In our region, this vaccine is only available in the private market and it is estimated a PCV13 vaccine coverage around 55% in children. A total of 1551 pneumococcal invasive isolates received between 2010 and 2013 in the Molecular Microbiology Department at Hospital Sant Joan de Déu, Barcelona, were included. Fifty-two serotypes and 249 clonal types-defined by MLST-were identified. The most common serotypes were serotype 1 (n = 182; 11.7%, 3 (n = 145; 9.3%, 19A (n = 137; 8.8% and 7F (n = 122; 7.9%. Serotype 14 was the third most frequent serotype in children < 2 years (15 of 159 isolates. PCV7 serotypes maintained their proportion along the period of study, 16.6% in 2010 to 13.4% in 2013, whereas there was a significant proportional decrease in PCV13 serotypes, 65.3% in 2010 to 48.9% in 2013 (p<0.01. This decrease was mainly attributable to serotypes 19A and 7F. Serotype 12F achieved the third position in 2013 (n = 22, 6.4%. The most frequent clonal types found were ST306 (n = 154, 9.9%, ST191 (n = 111, 7.2%, ST989 (n = 85, 5.5% and ST180 (n = 80, 5.2%. Despite their decrease, PCV13 serotypes continue to be a major cause of disease in Spain. These results emphasize the need for complete PCV13 vaccination.

  18. Clinical analysis of purulent meningitis caused by streptococcus pneumoniae in 12 children%儿童肺炎链球菌所致化脓性脑膜炎12例临床分析

    彭琴玲; 廖红梅; 唐静文; 陈玫; 杨赛

    2013-01-01

    目的 探讨肺炎链球菌所致的小儿化脓性脑膜炎的临床特点.方法 对我院2007年1月至2011年10月收治的12例肺炎链球菌所致的化脓性脑膜炎患儿的临床资料进行回顾性分析.结果 12例均为青霉素耐药肺炎链球菌,年龄2个月~9岁9个月,其中5岁以下占75% (9/12),2岁以下41.6%(5/12),临床表现均有发热,并伴有神经系统受累症状.12例患儿均有合并症,9例合并脓毒症(75%),8例合并肺炎(66.7%),其中5例同时合并脓毒症及肺炎.实验室检查外周血白细胞计数、C反应蛋白、降钙素原、红细胞沉降率及脑脊液白细胞计数、蛋白多明显增高,脑脊液糖下降明显.12例患儿中有11例患儿最终均使用万古霉素联合三、四代头孢或其他抗生素治疗,8例治愈,2例留有严重后遗症,2例死亡.结论 应重视肺炎链球菌所致的脑膜炎,对临床怀疑肺炎链球菌所致的化脓性脑膜炎,要尽早给予包括万古霉素在内的抗生素联合治疗.%Objective To investigate the clinical characteristics of purulent meningitis caused by streptococcus pneumoniae.Methods We studied clinical features of 12 children with purulent meningitis caused by streptococcus pneumoniae who were hospitalized from Jan 2007 to Oct 2011 in our hospital.Results Twelve children were penicillin-resistant streptococcus pneumoniae.The ages ranged from 2 months to 10 years.Nine cases(75%) were aged less than 5 years,and 5 cases(41.6%) were aged less than 2 years.All cases were with fever,and with the nervous system symptoms involvement.All cases were with complications:9 cases(75%) with septicemia,8 cases (66.7%) with pneumonia.The white blood cells,blood C-reactive protein,peripheral blood procalcitonin,erythrocyte sedimentation rate,white cells and protein in cerebrospinal fluid were mostly significantly high,sugar in cerebrospinal fluid were low significantly.Eleven cases were eventually treatmented by vancomycin plus 3rd

  19. Molecular characterization and antimicrobial susceptibility of Streptococcus pneumoniae isolated from children hospitalized with respiratory infections in Suzhou, China.

    Qian Geng

    Full Text Available BACKGROUND: Dissemination of antibiotic resistant clones is recognized as an important factor in the emergence and prevalence of resistance in pneumococcus. This study was undertaken to survey the antimicrobial susceptibility and serotypes distribution of pneumococci and to explore the circulating clones in hospitalized children in Suzhou, China. METHODS: The pneumococci were isolated from the nasopharyngeal aspirates of children less than 5 years of age admitted to Soochow-University-Affiliated-Children's-Hospital with respiratory infections. The capsular serotypes were identified by multiplex polymerase chain reaction (PCR. Antimicrobial susceptibility was tested by E-test. The presence of ermB, mefA/E genes were detected by PCR and the genotypes were explored by Multilocus sequence typing (MLST. RESULTS: From July 2012 to July 2013, a total of 175 pneumococcal isolates were collected and all strains were resistant to erythromycin and clindamycin, about 39.4% strains were non-susceptible to penicillin G. Overall, 174 (99.4% isolates were resistant to ≥ 3 types of antibiotics. Serotypes 19F (28.1%, 6B (19.7%, 19A (18.0%, and 23F (17.4% were the most common serotypes in all identified strains. The serotypes coverage of PCV7 and PCV13 were 71.9% and 89.9%, respectively. Four international antibiotic-resistant clones, including Taiwan19F-14 (n = 79, Spain23F-1(n = 25, Taiwan23F-15(n = 7 and Spain6B-2(n = 7, were identified. The Taiwan19F-14 clones have a higher non-susceptibility rate in β-lactams than other clones and non-clone isolates (p<0.001. In addition, 98.7% Taiwan19F-14 clones were positive of both ermB and mefA/E genes, compare to 33.3% in other clones and non-clone strains. CONCLUSIONS: The spread of international antibiotic-resistant clones, especially Taiwan19F-14 clones, played a predominant role in the dissemination of antimicrobial resistant isolates in Suzhou, China. Considering the high prevalence of PCV7 serotypes and

  20. Vigilancia epidemiológica centinela de Haemophilus influenzae y Streptococcus pneumoniae en menores de 5 años en el Perú

    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.