{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T16:50:25Z","timestamp":1791996625025,"version":"4.4.0"},"reference-count":37,"publisher":"MDPI AG","issue":"8","license":[{"start":{"date-parts":[[2024,8,12]],"date-time":"2024-08-12T00:00:00Z","timestamp":1723420800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003762","name":"Institut Pasteur","doi-asserted-by":"publisher","award":["24519E"],"award-info":[{"award-number":["24519E"]}],"id":[{"id":"10.13039\/501100003762","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Antibiotics"],"abstract":"<jats:p>Infections due to Haemophilus influnezae require prompt treatment using beta-lactam antibiotics. We used a collection of 81 isolates obtained between 1940 and 2001 from several countries. Whole genome sequencing showed the high heterogeneity of these isolates but allowed us to track the acquisition of beta-lactamase, which was first detected in 1980. Modifications of the ftsI gene encoding the penicillin-binding protein 3, PBP3, also involved in resistance to beta-lactams, appeared in 1991. These modifications (G490E, A502V, R517H, and N526K) were associated with resistance to amoxicillin that was not relieved by a beta-lactamase inhibitor (clavulanic acid), but the isolates retained susceptibility to third-generation cephalosporins (3GC). The modeling of the PBP3 structure suggested that these modifications may reduce the accessibility to the PBP3 active site. Other modifications appeared in 1998 and were associated with resistance to 3GC (S357N, M377I, S385T, and L389F). Modeling of the PBP3 structure suggested that they lie near the S379xN motif of the active site of PBP3. Overall resistance to amoxicillin was detected among 25 isolates (30.8%) of this collection. Resistance to sulfonamides was predicted by a genomic approach from the sequences of the folP gene (encoding the dihydropteroate synthase) due to difficulties in interpreting phenotypic anti-microbial testing and found in 13 isolates (16.0%). Our data suggest a slower spread of resistance to sulfonamides, which may be used for the treatment of H. influnezae infections. Genomic analysis may help in the prediction of antibiotic resistance, inform structure\u2013function analysis, and guide the optimal use of antibiotics.<\/jats:p>","DOI":"10.3390\/antibiotics13080761","type":"journal-article","created":{"date-parts":[[2024,8,13]],"date-time":"2024-08-13T13:35:49Z","timestamp":1723556149000},"page":"761","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["A Hunt for the Resistance of Haemophilus influnezae to Beta-Lactams"],"prefix":"10.3390","volume":"13","author":[{"given":"M\u00e9lanie","family":"Denizon","sequence":"first","affiliation":[{"name":"Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, Invasive Bacterial Infections Unit and National Reference Centre for Meningococci and Haemophilus influenzae, 28 Rue du Dr. Roux, CEDEX 15, 75724 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Hong","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, Invasive Bacterial Infections Unit and National Reference Centre for Meningococci and Haemophilus influenzae, 28 Rue du Dr. Roux, CEDEX 15, 75724 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aude","family":"Terrade","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, Invasive Bacterial Infections Unit and National Reference Centre for Meningococci and Haemophilus influenzae, 28 Rue du Dr. Roux, CEDEX 15, 75724 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0716-3174","authenticated-orcid":false,"given":"Muhamed-Kheir","family":"Taha","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, Invasive Bacterial Infections Unit and National Reference Centre for Meningococci and Haemophilus influenzae, 28 Rue du Dr. Roux, CEDEX 15, 75724 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2887-1732","authenticated-orcid":false,"given":"Ala-Eddine","family":"Deghmane","sequence":"additional","affiliation":[{"name":"Institut Pasteur, Universit\u00e9 Paris Cit\u00e9, Invasive Bacterial Infections Unit and National Reference Centre for Meningococci and Haemophilus influenzae, 28 Rue du Dr. Roux, CEDEX 15, 75724 Paris, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,8,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"2970","DOI":"10.1111\/j.1462-2920.2009.01972.x","article-title":"The role of antibiotics and antibiotic resistance in nature","volume":"11","author":"Aminov","year":"2009","journal-title":"Environ. Microbiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"25513","DOI":"10.1039\/C9CP00152B","article-title":"Reply to the \u2018Comment on \u201cPenicillin\u2019s catalytic mechanism revealed by inelastic neutrons and quantum chemical theory\u201d\u2019 by S. A. Glover Phys. Chem. Chem. Phys. 2019, 21, 18012","volume":"21","author":"Mucsi","year":"2019","journal-title":"Phys. Chem. Chem. Phys."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1111\/j.1574-6976.1999.tb00393.x","article-title":"Molecular determinants of the pathogenesis of disease due to non-typable Haemophilus influenzae","volume":"23","author":"Rao","year":"1999","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"S34","DOI":"10.1093\/infdis\/165-Supplement_1-S34","article-title":"Haemophilus influenzae Study Group Invasive Haemophilus influenzae disease: A population-based evaluation of the role of capsular polysaccharide serotype","volume":"165","author":"Wenger","year":"1992","journal-title":"J. Infect. Dis."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1093\/clinids\/12.1.75","article-title":"Global genetic structure and molecular epidemiology of encapsulated Haemophilus influenzae","volume":"12","author":"Musser","year":"1990","journal-title":"Rev. Infect. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1016\/S0031-3955(05)70210-8","article-title":"Update on Haemophilus influenzae serotype b and meningococcal vaccines","volume":"47","author":"Rosenstein","year":"2000","journal-title":"Pediatr. Clin. North. Am."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1715","DOI":"10.1093\/cid\/cit579","article-title":"Invasive Haemophilus influenzae type b disease in England and Wales: Who is at risk after 2 decades of routine childhood vaccination?","volume":"57","author":"Collins","year":"2013","journal-title":"Clin. Infect. Dis."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"6615","DOI":"10.1016\/j.vaccine.2018.09.060","article-title":"Increasing trend in invasive non-typeable Haemophilus influenzae disease and molecular characterization of the isolates, Italy, 2012\u20132016","volume":"36","author":"Fabiani","year":"2018","journal-title":"Vaccine"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1300","DOI":"10.1016\/j.cell.2014.11.017","article-title":"Beta-lactam antibiotics induce a lethal malfunctioning of the bacterial cell wall synthesis machinery","volume":"159","author":"Cho","year":"2014","journal-title":"Cell"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1126\/science.125.3238.99","article-title":"Strominger, Mode of action of penicillin","volume":"125","author":"Park","year":"1957","journal-title":"Science"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.jinf.2019.05.007","article-title":"High diversity of invasive Haemophilus influenzae isolates in France and the emergence of resistance to third generation cephalosporins by alteration of ftsI gene","volume":"79","author":"Deghmane","year":"2019","journal-title":"J. Infect."},{"key":"ref_12","first-page":"1314","article-title":"Antibiotic susceptibility and molecular analysis of invasive Haemophilus influenzae in Canada, 2007 to 2014","volume":"72","author":"Tsang","year":"2017","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5359","DOI":"10.2147\/IDR.S424468","article-title":"Epidemic Trends and Biofilm Formation Mechanisms of Haemophilus influenzae: Insights into Clinical Implications and Prevention Strategies","volume":"16","author":"Xiao","year":"2023","journal-title":"Infect. Drug Resist."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1093\/jac\/38.4.615","article-title":"Antimicrobial resistance and epidemiological study of Haemophilus influenzae strains isolated in Portugal. The Multicentre Study Group","volume":"38","author":"Casin","year":"1996","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"6227","DOI":"10.1016\/j.vaccine.2015.09.082","article-title":"Carriage of Haemophilus influenzae in the oropharynx of young children and molecular epidemiology of the isolates after fifteen years of H. influenzae type b vaccination in Italy","volume":"33","author":"Daprai","year":"2015","journal-title":"Vaccine"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.ijid.2020.08.040","article-title":"Analysis of Haemophilus species in patients with respiratory tract infections in Yaounde, Cameroon","volume":"100","author":"Tchatchouang","year":"2020","journal-title":"Int. J. Infect. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1089\/mdr.2015.0189","article-title":"Contribution of PBP3 Substitutions and TEM-1, TEM-15, and ROB-1 Beta-Lactamases to Cefotaxime Resistance in Haemophilus influenzae and Haemophilus parainfluenzae","volume":"22","author":"Sondergaard","year":"2016","journal-title":"Microb. Drug Resist."},{"key":"ref_18","unstructured":"World Health Organization (2024). World Health Organization WHO Bacterial Priority Pathogens List, 2024: Bacterial Pathogens of Public Health Importance to Guide Research, Development and Strategies to Prevent and Control Antimicrobial Resistance, WHO."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2208","DOI":"10.1128\/AAC.46.7.2208-2218.2002","article-title":"Diversity of beta-lactam resistance-conferring amino acid substitutions in penicillin-binding protein 3 of Haemophilus influenzae","volume":"46","author":"Dabernat","year":"2002","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1128\/AAC.45.6.1693-1699.2001","article-title":"Association of amino acid substitutions in penicillin-binding protein 3 with beta-lactam resistance in beta-lactamase-negative ampicillin-resistant Haemophilus influenzae","volume":"45","author":"Ubukata","year":"2001","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"508","DOI":"10.1007\/s10156-012-0372-0","article-title":"Antimicrobial susceptibility and genetic characteristics of Haemophilus influenzae isolated from community-acquired respiratory tract infection patients in Shanghai City, China","volume":"18","author":"Qin","year":"2012","journal-title":"J. Infect. Chemother."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/j.diagmicrobio.2018.10.009","article-title":"Detection of beta-lactamase-negative ampicillin resistance in Haemophilus influenzae in Belgium","volume":"93","author":"Schotte","year":"2019","journal-title":"Diagn. Microbiol. Infect. Dis."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Jolley, K.A., and Maiden, M.C. (2010). BIGSdb: Scalable analysis of bacterial genome variation at the population level. BMC Bioinform., 11.","DOI":"10.1186\/1471-2105-11-595"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1934","DOI":"10.1128\/AAC.46.6.1934-1939.2002","article-title":"Sulfonamide resistance in Haemophilus influenzae mediated by acquisition of sul2 or a short insertion in chromosomal folP","volume":"46","author":"Enne","year":"2002","journal-title":"Antimicrob. Agents Chemother."},{"key":"ref_25","unstructured":"European Committee on Antimicrobial Susceptibility Testing (2024, April 15). Breakpoint Tables for Interpretation of MICs and Zone Diameters. Version 14.0. Available online: http:\/\/www.eucast.org."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1203","DOI":"10.1099\/00221287-136-7-1203","article-title":"Urogenital, maternal and neonatal isolates of Haemophilus influenzae: Identification of unusually virulent serologically non-typable clone families and evidence for a new Haemophilus species","volume":"136","author":"Quentin","year":"1990","journal-title":"J. Gen. Microbiol."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Andrzejczuk, S., Kosikowska, U., Chwiejczak, E., St\u0119pie\u0144-Py\u015bniak, D., and Malm, A. (2019). Prevalence of Resistance to beta-Lactam Antibiotics and bla Genes. Among Commensal Haemophilus parainfluenzae Isolates from Respiratory Microbiota in Poland. Microorganisms, 7.","DOI":"10.3390\/microorganisms7100427"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"222","DOI":"10.1016\/j.jgar.2022.09.008","article-title":"Haemophilus influenzae drug resistance in France from 2017 to 2021: Consideration for treatment of otitis media","volume":"31","author":"Taha","year":"2022","journal-title":"J. Glob. Antimicrob. Resist."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"234","DOI":"10.1111\/j.1574-6976.2008.00105.x","article-title":"The penicillin-binding proteins: Structure and role in peptidoglycan biosynthesis","volume":"32","author":"Sauvage","year":"2008","journal-title":"FEMS Microbiol. Rev."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3501","DOI":"10.1016\/j.jmb.2019.07.010","article-title":"Novel and Improved Crystal Structures of H. influenzae, E. coli and P. aeruginosa Penicillin-Binding Protein 3 (PBP3) and N. gonorrhoeae PBP2: Toward a Better. Understanding of beta-Lactam Target-Mediated Resistance","volume":"431","author":"Bellini","year":"2019","journal-title":"J. Mol. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"16322","DOI":"10.1021\/ja038445l","article-title":"A mechanism-based inhibitor targeting the DD-transpeptidase activity of bacterial penicillin-binding proteins","volume":"125","author":"Lee","year":"2003","journal-title":"J. Am. Chem. Soc."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"241","DOI":"10.1007\/s13205-013-0147-z","article-title":"In silico study on Penicillin derivatives and Cephalosporins for upper respiratory tract bacterial pathogens","volume":"4","author":"Kumar","year":"2014","journal-title":"3 Biotech"},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Zapun, A., Morlot, C., and Taha, M.K. (2016). Resistance to beta-Lactams in Neisseria ssp Due to Chromosomally Encoded Penicillin-Binding Proteins. Antibiotics, 5.","DOI":"10.3390\/antibiotics5040035"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3163","DOI":"10.1093\/jac\/dkaa318","article-title":"Genomic epidemiology of Neisseria gonorrhoeae elucidating the gonococcal antimicrobial resistance and lineages\/sublineages across Brazil, 2015\u20132016","volume":"75","author":"Golparian","year":"2020","journal-title":"J. Antimicrob. Chemother."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2382","DOI":"10.1128\/jcm.32.10.2382-2386.1994","article-title":"PCR for capsular typing of Haemophilus influenzae","volume":"32","author":"Falla","year":"1994","journal-title":"J. Clin. Microbiol."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1093\/molbev\/msj030","article-title":"Application of phylogenetic networks in evolutionary studies","volume":"23","author":"Huson","year":"2006","journal-title":"Mol. Biol. Evol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1605","DOI":"10.1002\/jcc.20084","article-title":"UCSF Chimera--a visualization system for exploratory research and analysis","volume":"25","author":"Pettersen","year":"2004","journal-title":"J. Comput. Chem."}],"container-title":["Antibiotics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2079-6382\/13\/8\/761\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T15:35:18Z","timestamp":1760110518000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2079-6382\/13\/8\/761"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,8,12]]},"references-count":37,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2024,8]]}},"alternative-id":["antibiotics13080761"],"URL":"https:\/\/doi.org\/10.3390\/antibiotics13080761","relation":{},"ISSN":["2079-6382"],"issn-type":[{"value":"2079-6382","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,8,12]]}}}