{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,14]],"date-time":"2026-02-14T05:14:08Z","timestamp":1771046048543,"version":"3.50.1"},"reference-count":30,"publisher":"Public Library of Science (PLoS)","issue":"7","license":[{"start":{"date-parts":[[2022,7,25]],"date-time":"2022-07-25T00:00:00Z","timestamp":1658707200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003354","name":"Garnett Passe and Rodney Williams Memorial Foundation","doi-asserted-by":"publisher","award":["AT"],"award-info":[{"award-number":["AT"]}],"id":[{"id":"10.13039\/501100003354","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["Investigator Grant 1174876"],"award-info":[{"award-number":["Investigator Grant 1174876"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p><jats:italic>Haemophilus influenzae<\/jats:italic>, <jats:italic>Streptococcus pneumoniae<\/jats:italic> and <jats:italic>Moraxella catarrhalis<\/jats:italic> are bacterial species which frequently co-colonise the nasopharynx, but can also transit to the middle ear to cause otitis media. Chronic otitis media is often associated with a polymicrobial infection by these bacteria. However, despite being present in polymicrobial infections, the molecular interactions between these bacterial species remain poorly understood. We have previously reported competitive interactions driven by pH and growth phase between <jats:italic>H<\/jats:italic>. <jats:italic>influenzae<\/jats:italic> and <jats:italic>S<\/jats:italic>. <jats:italic>pneumoniae<\/jats:italic>. In this study, we have revealed competitive interactions between the three otopathogens, which resulted in reduction of <jats:italic>H<\/jats:italic>. <jats:italic>influenzae<\/jats:italic> viability in co-culture with <jats:italic>S<\/jats:italic>. <jats:italic>pneumoniae<\/jats:italic> and in triple-species culture. Transcriptomic analysis by mRNA sequencing identified a central role of arginine in mediating these interactions. Arginine supplementation was able to increase <jats:italic>H<\/jats:italic>. <jats:italic>influenzae<\/jats:italic> survival in a dual-species environment with <jats:italic>S<\/jats:italic>. <jats:italic>pneumoniae<\/jats:italic>, and in a triple-species environment. Arginine was used by <jats:italic>H<\/jats:italic>. <jats:italic>influenzae<\/jats:italic> for ATP production, and levels of ATP generated in dual- and triple-species co-culture at early stages of growth were significantly higher than the combined ATP levels of single-species cultures. These results indicate a central role for arginine-mediated ATP production by <jats:italic>H<\/jats:italic>. <jats:italic>influenzae<\/jats:italic> in the polymicrobial community.<\/jats:p>","DOI":"10.1371\/journal.pone.0271912","type":"journal-article","created":{"date-parts":[[2022,7,25]],"date-time":"2022-07-25T17:26:03Z","timestamp":1658769963000},"page":"e0271912","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":8,"title":["The central role of arginine in Haemophilus influenzae survival in a polymicrobial environment with Streptococcus pneumoniae and Moraxella catarrhalis"],"prefix":"10.1371","volume":"17","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9920-0245","authenticated-orcid":true,"given":"Alexandra","family":"Tikhomirova","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peter S.","family":"Zilm","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claudia","family":"Trappetti","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James C.","family":"Paton","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2118-1651","authenticated-orcid":true,"given":"Stephen P.","family":"Kidd","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2022,7,25]]},"reference":[{"issue":"3","key":"pone.0271912.ref001","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/S1473-3099(10)70012-8","article-title":"Otitis media and its consequences: beyond the earache","volume":"10","author":"A Vergison","year":"2010","journal-title":"Lancet Infect Dis"},{"key":"pone.0271912.ref002","first-page":"0502","article-title":"Otitis media with effusion in children","volume":"2015","author":"I. Williamson","year":"2015","journal-title":"BMJ Clin Evidence"},{"issue":"11","key":"pone.0271912.ref003","doi-asserted-by":"crossref","first-page":"1344","DOI":"10.1001\/jama.1993.03510110084037","article-title":"Use of antibiotics in preventing recurrent acute otitis media and in treating otitis media with effusion: A meta-analytic attempt to resolve the brouhaha","volume":"270","author":"RL Williams","year":"1993","journal-title":"JAMA"},{"key":"pone.0271912.ref004","doi-asserted-by":"crossref","first-page":"e20170125","DOI":"10.1542\/peds.2017-0125","article-title":"Effectiveness of Tympanostomy Tubes for Otitis Media: A Meta-analysis","author":"DW Steele","year":"2017","journal-title":"Pediatrics"},{"issue":"1","key":"pone.0271912.ref005","first-page":"1","volume":"13","author":"HC Smith-Vaughan","year":"2013","journal-title":"Dominance of Haemophilus influenzae in ear discharge from Indigenous Australian children with acute otitis media with tympanic membrane perforation"},{"issue":"2","key":"pone.0271912.ref006","doi-asserted-by":"crossref","first-page":"202","DOI":"10.1001\/jama.296.2.202","article-title":"Direct detection of bacterial biofilms on the middle-ear mucosa of children with chronic otitis media","volume":"296","author":"L Hall-Stoodley","year":"2006","journal-title":"JAMA"},{"issue":"7","key":"pone.0271912.ref007","doi-asserted-by":"crossref","first-page":"1045","DOI":"10.1093\/cid\/cix941","article-title":"Dynamics of bacterial colonization with Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis during symptomatic and asymptomatic viral upper respiratory tract infection","volume":"66","author":"GP DeMuri","year":"2018","journal-title":"Clin Infect Dis"},{"issue":"12","key":"pone.0271912.ref008","doi-asserted-by":"crossref","first-page":"e0167725","DOI":"10.1371\/journal.pone.0167725","article-title":"Increased nasopharyngeal density and concurrent carriage of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis are associated with pneumonia in febrile children","volume":"11","author":"S Chochua","year":"2016","journal-title":"PLOS One"},{"issue":"8","key":"pone.0271912.ref009","doi-asserted-by":"crossref","first-page":"881","DOI":"10.1016\/j.ijmm.2015.09.003","article-title":"The outcome of H. influenzae and S. pneumoniae inter-species interactions depends on pH, nutrient availability and growth phase","volume":"305","author":"A Tikhomirova","year":"2015","journal-title":"Int J Med Microbiol"},{"issue":"7","key":"pone.0271912.ref010","doi-asserted-by":"crossref","first-page":"fty074","DOI":"10.1093\/femspd\/fty074","article-title":"Specific growth conditions induce a Streptococcus pneumoniae non-mucoidal, small colony variant and determine the outcome of its co-culture with Haemophilus influenzae","volume":"76","author":"A Tikhomirova","year":"2018","journal-title":"Pathog Dis"},{"issue":"2","key":"pone.0271912.ref011","doi-asserted-by":"crossref","first-page":"114","DOI":"10.1111\/2049-632X.12073","article-title":"Haemophilus influenzae and Streptococcus pneumoniae: living together in a biofilm","volume":"69","author":"A Tikhomirova","year":"2013","journal-title":"Pathog Dis"},{"issue":"9","key":"pone.0271912.ref012","doi-asserted-by":"crossref","first-page":"705","DOI":"10.1080\/23744235.2018.1463451","article-title":"Antibody responses against Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis in children with acute respiratory infection with or without nasopharyngeal bacterial carriage","volume":"50","author":"DC Andrade","year":"2018","journal-title":"Infect Dis"},{"issue":"13","key":"pone.0271912.ref013","doi-asserted-by":"crossref","first-page":"2458","DOI":"10.1016\/j.vaccine.2006.09.020","article-title":"Modelling the co-occurrence of Streptococcus pneumoniae with other bacterial and viral pathogens in the upper respiratory tract","volume":"25","author":"P Jacoby","year":"2007","journal-title":"Vaccine"},{"issue":"10","key":"pone.0271912.ref014","doi-asserted-by":"crossref","first-page":"1584","DOI":"10.3201\/eid1410.080119","article-title":"Microbial interactions during upper respiratory tract infections","volume":"14","author":"MM Pettigrew","year":"2008","journal-title":"Emerging Infect Dis"},{"issue":"4","key":"pone.0271912.ref015","doi-asserted-by":"crossref","first-page":"630.e1","DOI":"10.1016\/j.cmi.2020.04.033","article-title":"Decreased prevalence of Moraxella catarrhalis in addition to Streptococcus pneumoniae in children with upper respiratory tract infection after introduction of conjugated pneumococcal vaccine: a retrospective cohort study","volume":"27","author":"N Littorin","year":"2021","journal-title":"Clin Microbiol Infect"},{"issue":"7","key":"pone.0271912.ref016","doi-asserted-by":"crossref","first-page":"1152","DOI":"10.1093\/infdis\/jit289","article-title":"Mixed pneumococcal\u2013nontypeable Haemophilus influenzae otitis media is a distinct clinical entity with unique epidemiologic characteristics and pneumococcal serotype distribution","volume":"208","author":"R Dagan","year":"2013","journal-title":"J Infect Dis"},{"issue":"5","key":"pone.0271912.ref017","doi-asserted-by":"crossref","first-page":"545","DOI":"10.1016\/j.micinf.2009.03.001","article-title":"The incidence of Streptococcus pneumoniae otitis media is affected by the polymicrobial environment particularly Moraxella catarrhalis in a mouse nasal colonisation model","volume":"11","author":"A Krishnamurthy","year":"2009","journal-title":"Microbes Infec"},{"issue":"8","key":"pone.0271912.ref018","doi-asserted-by":"crossref","first-page":"3845","DOI":"10.1128\/AAC.01772-10","article-title":"Moraxella catarrhalis outer membrane vesicles carry \u03b2-lactamase and promote survival of Streptococcus pneumoniae and Haemophilus influenzae by inactivating amoxicillin","volume":"55","author":"V Schaar","year":"2011","journal-title":"Antimicrobial Agents Chemother"},{"issue":"1","key":"pone.0271912.ref019","doi-asserted-by":"crossref","first-page":"218","DOI":"10.1186\/s13568-017-0521-z","article-title":"Inhibitory effect of Streptococci on the growth of M. catarrhalis strains and the diversity of putative bacteriocin-like gene loci in the genomes of S. pneumoniae and its relatives","volume":"7","author":"L Ikryannikova","year":"2017","journal-title":"AMB Express"},{"issue":"1","key":"pone.0271912.ref020","doi-asserted-by":"crossref","first-page":"ftaa077","DOI":"10.1093\/femspd\/ftaa077","article-title":"A single nucleotide polymorphism in an IgA1 protease gene determines Streptococcus pneumoniae adaptation to the middle ear during otitis media","volume":"79","author":"A Tikhomirova","year":"2021","journal-title":"Pathog Dis"},{"key":"pone.0271912.ref021","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1016\/j.fm.2019.01.007","article-title":"Morphophysiological responses of detached and adhered biofilms of Pseudomonas fluorescens to acidic electrolyzed water","volume":"82","author":"L-l Cai","year":"2019","journal-title":"Food Microbiol"},{"issue":"1","key":"pone.0271912.ref022","doi-asserted-by":"crossref","first-page":"e00180","DOI":"10.1128\/JB.00180-20","article-title":"Site-Specific mutations of GalR affect galactose metabolism in Streptococcus pneumoniae","volume":"203","author":"KT McLean","year":"2020","journal-title":"J Bacteriol"},{"issue":"3","key":"pone.0271912.ref023","doi-asserted-by":"crossref","first-page":"279","DOI":"10.1016\/S0960-9822(02)00478-5","article-title":"Metabolism and evolution of Haemophilus influenzae deduced from a whole-genome comparison with Escherichia coli","volume":"6","author":"RL Tatusov","year":"1996","journal-title":"Curr Biol"},{"issue":"10","key":"pone.0271912.ref024","doi-asserted-by":"crossref","first-page":"4959","DOI":"10.1128\/IAI.00073-07","article-title":"Metabolic analysis of Moraxella catarrhalis and the effect of selected in vitro growth conditions on global gene expression","volume":"75","author":"W Wang","year":"2007","journal-title":"Infect Immun"},{"issue":"52","key":"pone.0271912.ref025","doi-asserted-by":"crossref","first-page":"44594","DOI":"10.1074\/jbc.M111.295832","article-title":"Regulation of arginine acquisition and virulence gene expression in the human pathogen Streptococcus pneumoniae by transcription regulators ArgR1 and AhrC","volume":"286","author":"TG Kloosterman","year":"2011","journal-title":"J Biol Chem"},{"key":"pone.0271912.ref026","doi-asserted-by":"crossref","first-page":"42","DOI":"10.1186\/1471-2180-14-42","article-title":"Arginine deiminase pathway is far more important than urease for acid resistance and intracellular survival in Laribacter hongkongensis: a possible result of arc gene cassette duplication","volume":"14","author":"L Xiong","year":"2014","journal-title":"BMC Microbiol"},{"issue":"3","key":"pone.0271912.ref027","doi-asserted-by":"crossref","first-page":"429","DOI":"10.1016\/S1055-7903(02)00277-4","article-title":"Evolution of arginine deiminase (ADI) pathway genes","volume":"25","author":"M Z\u00fa\u00f1iga","year":"2002","journal-title":"Mol Phylogenet Evol"},{"issue":"27","key":"pone.0271912.ref028","doi-asserted-by":"crossref","first-page":"6053","DOI":"10.1021\/bi200497y","article-title":"Structural basis for the catalytic mechanism of aspartate ammonia lyase","volume":"50","author":"G Fibriansah","year":"2011","journal-title":"Biochemistry"},{"issue":"10","key":"pone.0271912.ref029","doi-asserted-by":"crossref","first-page":"3646","DOI":"10.1128\/JB.00088-08","article-title":"Regulation of gene expression in a mixed-genus community: stabilized arginine biosynthesis in Streptococcus gordonii by coaggregation with Actinomyces naeslundii","volume":"190","author":"NS Jakubovics","year":"2008","journal-title":"J Bacteriol"},{"issue":"35","key":"pone.0271912.ref030","doi-asserted-by":"crossref","first-page":"21185","DOI":"10.1074\/jbc.M115.644401","article-title":"Arginine-ornithine antiporter ArcD controls arginine metabolism and interspecies biofilm development of Streptococcus gordonii","volume":"290","author":"A Sakanaka","year":"2015","journal-title":"J Biol Chem"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0271912","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,7,25]],"date-time":"2022-07-25T17:26:42Z","timestamp":1658770002000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0271912"}},"subtitle":[],"editor":[{"given":"Hendrik W.","family":"van Veen","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2022,7,25]]},"references-count":30,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2022,7,25]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0271912","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,7,25]]}}}