Haemophilus influenzae carriage and antibiotic resistance profile in Belgian infants over a three-year period (2016–2018)

Background Non-typeable Haemophilus influenzae has become increasingly important as a causative agent of invasive diseases following vaccination against H. influenzae type b. The emergence of antibiotic resistance underscores the necessity to investigate typeable non-b carriage and non-typeable H. influenzae (NTHi) in children. Methods Nasopharyngeal swab samples were taken over a three-year period (2016–2018) from 336 children (6–30 months of age) attending daycare centers (DCCs) in Belgium, and from 218 children with acute otitis media (AOM). Biotype, serotype, and antibiotic resistance of H. influenzae strains were determined phenotypically. Mutations in the ftsI gene were explored in 129 strains that were resistant or had reduced susceptibility to beta-lactam antibiotics. Results were compared with data obtained during overlapping time periods from 94 children experiencing invasive disease. Results Overall, NTHi was most frequently present in both carriage (DCC, AOM) and invasive group. This was followed by serotype “f” (2.2%) and “e” (1.4%) in carriage, and “b” (16.0%), “f” (11.7%), and “a” (4.3%) in invasive strains. Biotype II was most prevalent in all studied groups, followed by biotype III in carriage and I in invasive strains. Strains from both groups showed highest resistance to ampicillin (26.7% in carriage vs. 18.1% in invasive group). A higher frequency of ftsI mutations were found in the AOM group than the DCC group (21.6 vs. 14.9% – p = 0.056). Even more so, the proportion of biotype III strains that carried a ftsI mutation was higher in AOM compared to DCC (50.0 vs. 26.3% – p < 0.01) and invasive group. Conclusion In both groups, NTHi was most frequently circulating, while specific encapsulated serotypes for carriage and invasive group were found. Biotypes I, II and III were more frequently present in the carriage and invasive group. The carriage group had a higher resistance-frequency to the analyzed antibiotics than the invasive group. Interestingly, a higher degree of ftsI mutations was found in children with AOM compared to DCC and invasive group. This data helps understanding the H. influenzae carriage in Belgian children, as such information is scarce.

[1]  B. Bozdoğan,et al.  Use of trans-complementation method to determine the effects of various ftsI mutations on β-lactamase-negative ampicillin-resistant (BLNAR) Haemophilus influenzae strains , 2022, Archives of Microbiology.

[2]  H. Goossens,et al.  Serotype 19A and 6C Account for One-Third of Pneumococcal Carriage Among Belgian Day-Care Children Four Years After a Shift to a Lower-Valent PCV , 2022, Journal of the Pediatric Infectious Diseases Society.

[3]  R. Drew,et al.  Haemophilus influenzae type f in the post-Haemophilus influenzae type b vaccination era: a systematic review. , 2022, Journal of medical microbiology.

[4]  M. Ulanova,et al.  Invasive Haemophilus influenzae Infections after 3 Decades of Hib Protein Conjugate Vaccine Use , 2021, Clinical microbiology reviews.

[5]  H. Claus,et al.  Cefotaxime resistance in invasive Haemophilus influenzae isolates in Germany 2016-19: prevalence, epidemiology and relevance of PBP3 substitutions. , 2021, Journal of Antimicrobial Chemotherapy.

[6]  N. Ahmad,et al.  Carriage of Haemophilus influenzae among children attending childcare centres in Kuala Lumpur, Malaysia in the post vaccination era: A cross-sectional study. , 2020, Vaccine.

[7]  R. Cunney,et al.  Epidemiology of Haemophilus influenzae in the Republic of Ireland, 2010–2018 , 2020, European Journal of Clinical Microbiology & Infectious Diseases.

[8]  W. Schaffner,et al.  Epidemiology of Invasive Haemophilus influenzae Serotype a Disease-United States, 2008-2017. , 2020, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[9]  E. Tanaka,et al.  Emergence of Haemophilus influenzae with low susceptibility to quinolones and persistence in tosufloxacin treatment. , 2019, Journal of global antimicrobial resistance.

[10]  Hong Zhang,et al.  Antibiotic Resistance Profiles of Haemophilus influenzae Isolates from Children in 2016: A Multicenter Study in China , 2019, The Canadian journal of infectious diseases & medical microbiology = Journal canadien des maladies infectieuses et de la microbiologie medicale.

[11]  K. Jolley,et al.  High diversity of invasive Haemophilus influenzae isolates in France and the emergence of resistance to third generation cephalosporins by alteration of ftsI gene. , 2019, The Journal of infection.

[12]  E. Choi,et al.  Increasing Prevalence of Group III Penicillin-Binding Protein 3 Mutations Conferring High-Level Resistance to Beta-Lactams Among Nontypeable Haemophilus influenzae Isolates from Children in Korea. , 2019, Microbial drug resistance.

[13]  Lise Schotte,et al.  Detection of beta-lactamase-negative ampicillin resistance in Haemophilus influenzae in Belgium. , 2019, Diagnostic microbiology and infectious disease.

[14]  Hiroki Takahashi,et al.  Multiclonal Expansion and High Prevalence of β-Lactamase-Negative Haemophilus influenzae with High-Level Ampicillin Resistance in Japan and Susceptibility to Quinolones , 2018, Antimicrobial Agents and Chemotherapy.

[15]  H. Goossens,et al.  Nasopharyngeal s. pneumoniae carriage and density in Belgian infants after 9 years of pneumococcal conjugate vaccine programme. , 2018, Vaccine.

[16]  F. Ruiz-López,et al.  Epidemiology of Nasopharyngeal Carriage by Haemophilus influenzae in Healthy Children: A Study in the Mediterranean Coast Region , 2017, The Pediatric infectious disease journal.

[17]  M. Ulanova,et al.  The changing epidemiology of invasive Haemophilus influenzae disease: Emergence and global presence of serotype a strains that may require a new vaccine for control. , 2017, Vaccine.

[18]  M. Slack The evidence for non-typeable Haemophilus influenzae as a causative agent of childhood pneumonia , 2017, Pneumonia.

[19]  D. Kunde,et al.  Effect of epithelial cell type on in vitro invasion of non-typeable Haemophilus influenzae. , 2016, Journal of microbiological methods.

[20]  E. Redwan,et al.  Hib Vaccines: Past, Present, and Future Perspectives , 2016, Journal of immunology research.

[21]  S. Ladhani,et al.  Non-typeable Haemophilus influenzae, an under-recognised pathogen. , 2014, The Lancet. Infectious diseases.

[22]  G. Zosky,et al.  Genotypically defined β-lactamase-negative ampicillin-resistant isolates of non-typable Haemophilus influenzae are associated with increased invasion of bronchial epithelial cells in vitro. , 2014, Journal of medical microbiology.

[23]  A. Sundsfjord,et al.  Multilocus sequence typing and ftsI sequencing: a powerful tool for surveillance of penicillin-binding protein 3-mediated beta-lactam resistance in nontypeable Haemophilus influenzae , 2014, BMC Microbiology.

[24]  S. Ladhani,et al.  Invasive Haemophilus influenzae Serotype e and f Disease, England and Wales , 2012, Emerging infectious diseases.

[25]  T. Murphy,et al.  Haemophilus influenzae Infections in the H. influenzae Type b Conjugate Vaccine Era , 2011, Journal of Clinical Microbiology.

[26]  M. Cerquetti,et al.  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. , 2011, The Journal of antimicrobial chemotherapy.

[27]  E. Kasuga,et al.  An amino acid substitution in PBP-3 in Haemophilus influenzae associate with the invasion to bronchial epithelial cells. , 2010, Microbiological research.

[28]  Hideaki Suzuki,et al.  Nasopharyngeal Haemophilus influenzae Carriage in Japanese Children Attending Day-Care Centers , 2008, Journal of Clinical Microbiology.

[29]  N. Yamanaka,et al.  Genetic Characteristics and Clonal Dissemination of β-Lactamase-Negative Ampicillin-Resistant Haemophilus influenzae Strains Isolated from the Upper Respiratory Tract of Patients in Japan , 2007, Antimicrobial Agents and Chemotherapy.

[30]  J. Campos,et al.  Ampicillin-Resistant Non-β-Lactamase-Producing Haemophilus influenzae in Spain: Recent Emergence of Clonal Isolates with Increased Resistance to Cefotaxime and Cefixime , 2007, Antimicrobial Agents and Chemotherapy.

[31]  G. Swingler,et al.  Conjugate vaccines for preventing Haemophilus influenzae type B infections. , 2007, The Cochrane database of systematic reviews.

[32]  S. Awasthi,et al.  High ampicillin resistance in different biotypes and serotypes of Haemophilus influenzae colonizing the nasopharynx of healthy school-going Indian children. , 2006, Journal of medical microbiology.

[33]  Sabine Mérienne,et al.  Programme , 1953, Neuromuscular Disorders.

[34]  K. Sunakawa,et al.  Diversity of ampicillin-resistance genes in Haemophilus influenzae in Japan and the United States. , 2003, Microbial drug resistance.

[35]  K. Sunakawa,et al.  Differentiation of β-lactamase-negative ampicillin-resistant Haemophilus influenzae from other H. influenzae strains by a disc method , 2002, Journal of infection and chemotherapy : official journal of the Japan Society of Chemotherapy.

[36]  K. Yamamoto,et al.  Association of Amino Acid Substitutions in Penicillin-Binding Protein 3 with β-Lactam Resistance in β-Lactamase-Negative Ampicillin-Resistant Haemophilus influenzae , 2001, Antimicrobial Agents and Chemotherapy.

[37]  H. Peltola Worldwide Haemophilus influenzae type b disease at the beginning of the 21st century: global analysis of the disease burden 25 years after the use of the polysaccharide vaccine and a decade after the advent of conjugates. , 2000, Clinical microbiology reviews.

[38]  D. Madore Impact of immunization on Haemophilus influenzae type b disease. , 1996, Infectious agents and disease.

[39]  C. Svanborg,et al.  Nasopharyngeal colonization during the first year of life. , 1992, The Journal of infectious diseases.

[40]  J. Righter,et al.  Haemophilus influenzae from four laboratories in one Canadian city. , 1988, The Journal of antimicrobial chemotherapy.

[41]  K C Watson,et al.  Temporal changes in biotypes of Haemophilus influenzae isolated from patients with cystic fibrosis. , 1988, Journal of medical microbiology.

[42]  J. Musser,et al.  Genetic and phenotypic diversity among ampicillin-resistant, non-beta-lactamase-producing, nontypeable Haemophilus influenzae isolates , 1987, Infection and immunity.

[43]  G. Jacoby,et al.  An animal source for the ROB-1 beta-lactamase of Haemophilus influenzae type b , 1986, Antimicrobial Agents and Chemotherapy.

[44]  P. Gilligan,et al.  Biotype of Haemophilus influenzae: correlation with virulence and ampicillin resistance. , 1983, The Journal of infectious diseases.

[45]  B. Mittelman,et al.  On control. , 1979, Dental management.

[46]  J. Brunton,et al.  Biochemical characteristics of Haemophilus influenzae in relationship to source of isolation and antibiotic resistance , 1978, Journal of clinical microbiology.

[47]  P. Mäkelä,et al.  Haemophilus influenzae type b capsular polysaccharide vaccine in children: a double-blind field study of 100,000 vaccinees 3 months to 5 years of age in Finland. , 1977, Pediatrics.

[48]  M. Kilian A taxonomic study of the genus Haemophilus, with the proposal of a new species. , 1976, Journal of general microbiology.

[49]  M. Pittman VARIATION AND TYPE SPECIFICITY IN THE BACTERIAL SPECIES HEMOPHILUS INFLUENZAE , 1931, The Journal of experimental medicine.