Characteristics of Haemophilus influenzae invasive isolates from Portugal following routine childhood vaccination against H. influenzae serotype b (2002–2010)

  • M. P. Bajanca-Lavado
  • A. S. Simões
  • C. R. Betencourt
  • R. Sá-Leão
  • The Portuguese Group for the Study of Haemophilus influenzae invasive infection
Article

Abstract

We aimed to characterize Haemophilus influenzae invasive isolates recovered in Portugal over a 9-year period (2002–2010) following the inclusion of H. influenzae serotype b (Hib) conjugate vaccination in the National Immunization Program (NIP) in the year 2000 and compare the results with those obtained in a similar study from the pre-vaccination era (1989–2001) previously described by us. As part of a laboratory-based passive surveillance system, 144 invasive isolates obtained in 28 Portuguese hospitals were received at the National Reference Laboratory for Bacterial Respiratory Infections and were characterized. Capsular types and antibiotic susceptibility patterns were determined. The ftsI gene encoding PBP3 was sequenced for β-lactamase-negative ampicillin-resistant (BLNAR) isolates. Genetic relatedness among isolates was examined by multilocus sequencing typing (MLST). Most isolates (77.1 %) were non-capsulated, a significant increase compared to the pre-vaccination era (19.0 %, p < 0.001). Serotype b strains decreased significantly (from 81.0 to 13.2 %, p < 0.001) and serotype f increased significantly (from 0.8 to 6.9 %, p = 0.03). Ten percent of the isolates were β-lactamase producers, a value lower than that previously observed (26.9 %, p = 0.005). Eight percent of all isolates were BLNAR. A high genetic diversity among non-capsulated isolates was found. By contrast, capsulated isolates were clonal. The implementation of Hib vaccination has resulted in a significant decline in the proportion of serotype b H. influenzae invasive disease isolates. Most episodes of invasive disease occurring in Portugal are now due to fully susceptible, highly diverse, non-capsulated strains. Given the evolving dynamics of this pathogen and the increase in non-type b capsulated isolates, continuous surveillance is needed.

References

  1. 1.
    Watt JP, Wolfson LJ, O’Brien KL, Henkle E, Deloria-Knoll M, McCall N, Lee E, Levine OS, Hajjeh R, Mulholland K, Cherian T; Hib and Pneumococcal Global Burden of Disease Study Team (2009) Burden of disease caused by Haemophilus influenzae type b in children younger than 5 years: global estimates. Lancet 374(9693):903–911PubMedCrossRefGoogle Scholar
  2. 2.
    Rubach MP, Bender JM, Mottice S, Hanson K, Weng HY, Korgenski K, Daly JA, Pavia AT (2011) Increasing incidence of invasive Haemophilus influenzae disease in adults, Utah, USA. Emerg Infect Dis 17(9):1645–1650PubMedCentralPubMedCrossRefGoogle Scholar
  3. 3.
    Rubin LG, Medeiros AA, Yolken RH, Moxon ER (1981) Ampicillin treatment failure of apparently beta-lactamase-negative Haemophilus influenzae type b meningitis due to novel beta-lactamase. Lancet 2(8254):1008–1010PubMedCrossRefGoogle Scholar
  4. 4.
    Tristram S, Jacobs MR, Appelbaum PC (2007) Antimicrobial resistance in Haemophilus influenzae. Clin Microbiol Rev 20(2):368–389PubMedCentralPubMedCrossRefGoogle Scholar
  5. 5.
    Ubukata K, Shibasaki Y, Yamamoto K, Chiba N, Hasegawa K, Takeuchi Y, Sunakawa K, Inoue M, Konno M (2001) Association of amino acid substitutions in penicillin-binding protein 3 with beta-lactam resistance in beta-lactamase-negative ampicillin-resistant Haemophilus influenzae. Antimicrob Agents Chemother 45(6):1693–1699PubMedCentralPubMedCrossRefGoogle Scholar
  6. 6.
    Hasegawa K, Yamamoto K, Chiba N, Kobayashi R, Nagai K, Jacobs MR, Appelbaum PC, Sunakawa K, Ubukata K (2003) Diversity of ampicillin-resistance genes in Haemophilus influenzae in Japan and the United States. Microb Drug Resist 9(1):39–46PubMedCrossRefGoogle Scholar
  7. 7.
    Clinical and Laboratory Standards Institute (CLSI) (2011) Performance standards for antimicrobial susceptibility testing; Twenty-first informational supplement. CLSI document M100-S21. CLSI, Wayne, PA, USAGoogle Scholar
  8. 8.
    Tristram SG, Burdach JG (2007) Effect of cloned inhibitor-resistant TEM beta-lactamases on the susceptibility of Haemophilus influenzae to amoxicillin/clavulanate. J Antimicrob Chemother 60(5):1151–1154PubMedCrossRefGoogle Scholar
  9. 9.
    Meats E, Feil EJ, Stringer S, Cody AJ, Goldstein R, Kroll JS, Popovic T, Spratt BG (2003) Characterization of encapsulated and noncapsulated Haemophilus influenzae and determination of phylogenetic relationships by multilocus sequence typing. J Clin Microbiol 41(4):1623–1636PubMedCentralPubMedCrossRefGoogle Scholar
  10. 10.
    Bajanca P, Caniça M; Multicenter Study Group (2004) Emergence of nonencapsulated and encapsulated non-b-type invasive Haemophilus influenzae isolates in Portugal (1989–2001). J Clin Microbiol 42(2):807–810PubMedCentralPubMedCrossRefGoogle Scholar
  11. 11.
    Falla TJ, Crook DW, Brophy LN, Maskell D, Kroll JS, Moxon ER (1994) PCR for capsular typing of Haemophilus influenzae. J Clin Microbiol 32(10):2382–2386PubMedCentralPubMedGoogle Scholar
  12. 12.
    Mendonça N, Ferreira E, Caniça M (2006) Occurrence of a novel SHV-type enzyme (SHV-55) among isolates of Klebsiella pneumoniae from Portuguese origin in a comparison study for extended-spectrum beta-lactamase-producing evaluation. Diagn Microbiol Infect Dis 56(4):415–420PubMedCrossRefGoogle Scholar
  13. 13.
    Cerquetti M, Giufrè M, Cardines R, Mastrantonio P (2007) First characterization of heterogeneous resistance to imipenem in invasive nontypeable Haemophilus influenzae isolates. Antimicrob Agents Chemother 51(9):3155–3161PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Barbosa AR, Giufrè M, Cerquetti M, Bajanca-Lavado MP (2011) Polymorphism in ftsI gene and beta-lactam susceptibility in Portuguese Haemophilus influenzae strains: clonal dissemination of beta-lactamase-positive isolates with decreased susceptibility to amoxicillin/clavulanic acid. J Antimicrob Chemother 66(4):788–796PubMedCentralPubMedCrossRefGoogle Scholar
  15. 15.
    Dabernat H, Delmas C, Seguy M, Pelissier R, Faucon G, Bennamani S, Pasquier C (2002) Diversity of beta-lactam resistance-conferring amino acid substitutions in penicillin-binding protein 3 of Haemophilus influenzae. Antimicrob Agents Chemother 46(7):2208–2218PubMedCentralPubMedCrossRefGoogle Scholar
  16. 16.
    Livorsi DJ, Macneil JR, Cohn AC, Bareta J, Zansky S, Petit S, Gershman K, Harrison LH, Lynfield R, Reingold A, Schaffner W, Thomas A, Farley MM (2012) Invasive Haemophilus influenzae in the United States, 1999–2008: epidemiology and outcomes. J Infect 65(6):496–504PubMedCrossRefGoogle Scholar
  17. 17.
    Ulanova M, Tsang RS (2009) Invasive Haemophilus influenzae disease: changing epidemiology and host-parasite interactions in the 21st century. Infect Genet Evol 9(4):594–605PubMedCrossRefGoogle Scholar
  18. 18.
    Giufrè M, Cardines R, Caporali MG, Accogli M, D’Ancona F, Cerquetti M (2011) Ten years of Hib vaccination in Italy: prevalence of non-encapsulated Haemophilus influenzae among invasive isolates and the possible impact on antibiotic resistance. Vaccine 29(22):3857–3862PubMedCrossRefGoogle Scholar
  19. 19.
    Kastrin T, Paragi M, Kolman J, Cizman M, Kraigher A, Gubina M; Slovenian Meningitidis Study Group (2010) Characterisation of invasive Haemophilus influenzae isolates in Slovenia, 1993–2008. Eur J Clin Microbiol Infect Dis 29(6):661–668PubMedCrossRefGoogle Scholar
  20. 20.
    Laupland KB, Schønheyder HC, Østergaard C, Knudsen JD, Valiquette L, Galbraith J, Kennedy KJ, Gradel KO; International Bacteremia Surveillance Collaborative (2011) Epidemiology of Haemophilus influenzae bacteremia: a multi-national population-based assessment. J Infect 62(2):142–148PubMedCrossRefGoogle Scholar
  21. 21.
    Resman F, Ristovski M, Ahl J, Forsgren A, Gilsdorf JR, Jasir A, Kaijser B, Kronvall G, Riesbeck K (2012) Invasive disease caused by Haemophilus influenzae in Sweden 1997–2009; evidence of increasing incidence and clinical burden of non-type b strains. Clin Microbiol Infect 17(11):1638–1645CrossRefGoogle Scholar
  22. 22.
    Zanella RC, Bokermann S, Andrade AL, Flannery B, Brandileone MC (2011) Changes in serotype distribution of Haemophilus influenzae meningitis isolates identified through laboratory-based surveillance following routine childhood vaccination against H. influenzae type b in Brazil. Vaccine 29(48):8937–8942PubMedCrossRefGoogle Scholar
  23. 23.
    Ladhani SN, Collins S, Vickers A, Litt DJ, Crawford C, Ramsay ME, Slack MP (2012) Invasive Haemophilus influenzae serotype e and f disease, England and Wales. Emerg Infect Dis 18(5):725–732PubMedCentralPubMedCrossRefGoogle Scholar
  24. 24.
    Campos J, Hernando M, Román F, Pérez-Vázquez M, Aracil B, Oteo J, Lázaro E, de Abajo F; Group of Invasive Haemophilus Infections of the Autonomous Community of Madrid, Spain (2004) Analysis of invasive Haemophilus influenzae infections after extensive vaccination against H. influenzae type b. J Clin Microbiol 42(2):524–529PubMedCentralPubMedCrossRefGoogle Scholar
  25. 25.
    Bruun B, Gahrn-Hansen B, Westh H, Kilian M (2004) Clonal relationship of recent invasive Haemophilus influenzae serotype f isolates from Denmark and the United States. J Med Microbiol 53(Pt 11):1161–1165PubMedCrossRefGoogle Scholar
  26. 26.
    Calado R, Betencourt C, Gonçalves H, Cristino N, Calhau P, Lavado PB (2011) Complicated meningitis caused by a rare serotype of Haemophilus influenzae in Portugal. Diagn Microbiol Infect Dis 69(1):111–113PubMedCrossRefGoogle Scholar
  27. 27.
    Ladhani S, Slack MP, Heath PT, von Gottberg A, Chandra M, Ramsay ME; European Union Invasive Bacterial Infection Surveillance participants (2010) Invasive Haemophilus influenzae Disease, Europe, 1996–2006. Emerg Infect Dis 16(3):455–463PubMedCentralPubMedCrossRefGoogle Scholar
  28. 28.
    García-Cobos S, Campos J, Cercenado E, Román F, Lázaro E, Pérez-Vázquez M, de Abajo F, Oteo J (2008) Antibiotic resistance in Haemophilus influenzae decreased, except for beta-lactamase-negative amoxicillin-resistant isolates, in parallel with community antibiotic consumption in Spain from 1997 to 2007. Antimicrob Agents Chemother 52(8):2760–2766PubMedCentralPubMedCrossRefGoogle Scholar
  29. 29.
    Resman F, Ristovski M, Forsgren A, Kaijser B, Kronvall G, Medstrand P, Melander E, Odenholt I, Riesbeck K (2012) Increase of beta-lactam-resistant invasive Haemophilus influenzae in Sweden, 1997 to 2010. Antimicrob Agents Chemother 56(8):4408–4415PubMedCentralPubMedCrossRefGoogle Scholar
  30. 30.
    Shuel ML, Tsang RS (2009) Canadian beta-lactamase-negative Haemophilus influenzae isolates showing decreased susceptibility toward ampicillin have significant penicillin binding protein 3 mutations. Diagn Microbiol Infect Dis 63(4):379–383PubMedCrossRefGoogle Scholar
  31. 31.
    Hotomi M, Fujihara K, Billal DS, Suzuki K, Nishimura T, Baba S, Yamanaka N (2007) Genetic characteristics and clonal dissemination of beta-lactamase-negative ampicillin-resistant Haemophilus influenzae strains isolated from the upper respiratory tract of patients in Japan. Antimicrob Agents Chemother 51(11):3969–3976PubMedCentralPubMedCrossRefGoogle Scholar
  32. 32.
    García-Cobos S, Campos J, Román F, Carrera C, Pérez-Vázquez M, Aracil B, Oteo J (2008) Low beta-lactamase-negative ampicillin-resistant Haemophilus influenzae strains are best detected by testing amoxicillin susceptibility by the broth microdilution method. Antimicrob Agents Chemother 52(7):2407–2414PubMedCentralPubMedCrossRefGoogle Scholar
  33. 33.
    Jansen WT, Verel A, Beitsma M, Verhoef J, Milatovic D (2006) Longitudinal European surveillance study of antibiotic resistance of Haemophilus influenzae. J Antimicrob Chemother 58(4):873–877PubMedCrossRefGoogle Scholar
  34. 34.
    Sanbongi Y, Suzuki T, Osaki Y, Senju N, Ida T, Ubukata K (2006) Molecular evolution of beta-lactam-resistant Haemophilus influenzae: 9-year surveillance of penicillin-binding protein 3 mutations in isolates from Japan. Antimicrob Agents Chemother 50(7):2487–2492PubMedCentralPubMedCrossRefGoogle Scholar
  35. 35.
    Hasegawa K, Kobayashi R, Takada E, Ono A, Chiba N, Morozumi M, Iwata S, Sunakawa K, Ubukata K; Nationwide Surveillance for Bacterial Meningitis (2006) High prevalence of type b beta-lactamase-non-producing ampicillin-resistant Haemophilus influenzae in meningitis: the situation in Japan where Hib vaccine has not been introduced. J Antimicrob Chemother 57(6):1077–1082PubMedCrossRefGoogle Scholar
  36. 36.
    Sevillano D, Giménez MJ, Cercenado E, Cafini F, Gené A, Alou L, Marco F, Martínez-Martínez L, Coronel P, Aguilar L (2009) Genotypic versus phenotypic characterization, with respect to beta-lactam susceptibility, of Haemophilus influenzae isolates exhibiting decreased susceptibility to beta-lactam resistance markers. Antimicrob Agents Chemother 53(1):267–270PubMedCentralPubMedCrossRefGoogle Scholar
  37. 37.
    Melo-Cristino J, Fernandes ML, Serrano N; Portuguese Surveillance Group for the Study of Respiratory Pathogens (2001) A multicenter study of the antimicrobial susceptibility of Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis isolated from patients with community-acquired lower respiratory tract infections in 1999 in Portugal. Microb Drug Resist 7(1):33–38PubMedCrossRefGoogle Scholar
  38. 38.
    Bajanca-Lavado MP, Casin I, Vaz Pato MV (1996) Antimicrobial resistance and epidemiological study of Haemophilus influenzae strains isolated in Portugal. The Multicentre Study Group. J Antimicrob Chemother 38(4):615–625PubMedCrossRefGoogle Scholar
  39. 39.
    García-Cobos S, Arroyo M, Pérez-Vázquez M, Aracil B, Oteo J, Campos J (2012) Evaluation of the EUCAST disc diffusion susceptibility testing method for Haemophilus influenzae based on the resistance mechanism to beta-lactam antibiotics. J Antimicrob Chemother 68(1):159–163PubMedCrossRefGoogle Scholar
  40. 40.
    Søndergaard A, Petersen MT, Fuursted K, Nørskov-Lauritsen N (2012) Detection of N526K-substituted penicillin-binding protein 3 conferring low-level mutational resistance to beta-lactam antibiotics in Haemophilus influenzae by disc diffusion testing on Mueller–Hinton agar according to EUCAST guidelines. J Antimicrob Chemother 67(6):1401–1404PubMedCrossRefGoogle Scholar
  41. 41.
    Alou L, Giménez MJ, Sevillano D, Aguilar L, González N, Echeverría O, Torrico M, Coronel P, Prieto J (2007) Are beta-lactam breakpoints adequate to define non-susceptibility for all Haemophilus influenzae resistance phenotypes from a pharmacodynamic point of view? J Antimicrob Chemother 59(4):652–657PubMedCrossRefGoogle Scholar
  42. 42.
    García-de-Lomas J, Lerma M, Cebrián L, Juan-Bañón JL, Coronel P, Giménez MJ, Aguilar L (2007) Influence of Haemophilus influenzae beta-lactamase production and/or ftsI gene mutations on in vitro activity of and susceptibility rates to aminopenicillins and second- and third-generation cephalosporins. Int J Antimicrob Agents 30(2):190–192PubMedCrossRefGoogle Scholar
  43. 43.
    García-Cobos S, Campos J, Lázaro E, Román F, Cercenado E, García-Rey C, Pérez-Vázquez M, Oteo J, de Abajo F (2007) Ampicillin-resistant non-β-lactamase-producing Haemophilus influenzae in Spain: recent emergence of clonal isolates with increased resistance to cefotaxime and cefixime. Antimicrob Agents Chemother 51(7):2564–2573PubMedCentralPubMedCrossRefGoogle Scholar
  44. 44.
    Ubukata K, Chiba N, Morozumi M, Iwata S, Sunakawa K; Working Group of Nationwide Surveillance for Bacterial Meningitis (2013) Longitudinal surveillance of Haemophilus influenzae isolates from pediatric patients with meningitis throughout Japan, 2000–2011. J Infect Chemother 19(1):34–41PubMedCrossRefGoogle Scholar
  45. 45.
    Heilmann KP, Rice CL, Miller AL, Miller NJ, Beekmann SE, Pfaller MA, Richter SS, Doern GV (2005) Decreasing prevalence of beta-lactamase production among respiratory tract isolates of Haemophilus influenzae in the United States. Antimicrob Agents Chemother 49(6):2561–2564PubMedCentralPubMedCrossRefGoogle Scholar
  46. 46.
    Connor TR, Corander J, Hanage WP (2012) Population subdivision and the detection of recombination in non-typable Haemophilus influenzae. Microbiology 158(Pt 12):2958–2964PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • M. P. Bajanca-Lavado
    • 1
  • A. S. Simões
    • 2
  • C. R. Betencourt
    • 1
  • R. Sá-Leão
    • 2
  • The Portuguese Group for the Study of Haemophilus influenzae invasive infection
  1. 1.National Reference Laboratory for Bacterial Respiratory Infections, Department of Infectious DiseaseNational Institute of Health Dr. Ricardo JorgeLisboaPortugal
  2. 2.Laboratory of Molecular Microbiology of Human PathogensInstituto de Tecnologia Química e Biológica, Universidade Nova de LisboaOeirasPortugal

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