Non typable-Haemophilus influenzae biofilm formation and acute otitis media

BMC Infectious Diseases

Josette Raymond³  Corinne Levy²  Claire Poyart³  Stephane Bechet²  Stephane Bonacorsi¹  Emmanuelle Varon⁴  Robert Cohen²  Assaf Mizrahi³ 

[1] Université Diderot, Hôpital Robert Debré, Bactériologie, Paris, France;ACTIV, Saint Maur des Fossés, Paris, France;Université Paris Descartes, Hôpital Cochin, Bactériologie, 27 rue du Faubourg Saint Jacques, 75679 Paris cedex 14, France;Université Paris Descartes, Hôpital Georges Pompidou, Bactériologie, Paris, France

关键词: Conjunctivitis;    AOM;    Biofilm;    Haemophilus influenzae;   

Others  :  1127328
DOI  :  10.1186/1471-2334-14-400

 received in 2014-02-21, accepted in 2014-07-12,  发布年份 2014

【 摘 要 】

Background

Non-typable Haemophilus influenzae (NT-Hi) infection is frequently associated with acute otitis media (AOM) treatment failure, recurrence or chronic otitis media. Persistence of otopathogens in a biofilm-structured community was implicated in these situations. Here, we compared biofilm production by H. influenzae strains obtained by culture of middle ear fluid (MEF) from children with AOM treatment failure and by strains isolated from nasopharyngeal (NP) samples from healthy children or those with AOM (first episode or recurrence). We aimed to evaluate an association of clinical signs and in vitro biofilm formation and establish risk factors of carrying a biofilm-producing strain.

Methods

We used a modification of the microtiter plate assay with crystal violet staining to compare biofilm production by 216 H. influenzae strains: 41 in MEF from children with AOM treatment failure (group MEF), 43 in NP samples from healthy children (NP group 1), 88 in NP samples from children with a first AOM episode (NP group 2, n = 43) or recurrent (NP group 3, n = 45) and 44 in NP samples from children with AOM associated with conjunctivitis (NP group 4).

Results

At all, 106/216 (49%) H. influenzae strains produced biofilm as did 26/43 (60.5%) in NP samples from healthy children. Biofilm production in MEF samples and NP samples did not significantly differ (40.5% vs 60.5%, 55.8%, 56.8% and 31.1% for NP groups 1, 2, 3 and 4, respectively). On multivariate analysis, only presence of conjunctivitis was significantly associated with low biofilm production (OR = 0.3, CI [0.16-0.60], p = 0.001). The ampicillin resistance of H. influenzae produced by penicillin-binding protein modification was significantly associated with low biofilm production (p = 0.029).

Conclusion

We found no association of biofilm production and AOM treatment failure or recurrence. Biofilm production was low from H. influenzae strains associated with conjunctivitis-otitis syndrome and from strains with modified penicillin-binding protein.

【 授权许可】

   
2014 Mizrahi et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

【 参考文献 】

• [1]Casey JR, Pichichero ME: Changes in frequency and pathogens causing acute otitis media in 1995-2003. Pediatr Infect Dis J 2004, 23(9):824-828.
• [2]Block SL, Hedrick J, Harrison CJ, Tyler R, Smith A, Findlay R, Keegan E: Community-wide vaccination with the heptavalent pneumococcal conjugate significantly alters the microbiology of acute otitis media. Pediatr Infect Dis J 2004, 23(9):829-833.
• [3]Couloigner V, Levy C, François M, Bidet P, Hausdorff WP, Pascal T, Boucherat M, Bingen E, Mariani P, Pierrot S, Bille E, Carbonnelle E, Varon E, Cohen R: Pathogens implicated in acute otitis media failures after 7-valent pneumococcal conjugate vaccine implementation in France: distribution, serotypes, and resistance levels. Pediatr Infect Dis J 2012, 31(2):154-158.
• [4]Schwartz RH, Rodriguez WJ, Khan WN: Persistent purulent otitis media. Clin Pediatr (Phila) 1981, 20(7):445-447.
• [5]Barkai G, Leibovitz E, Givon-Lavi N, Dagan R: Potential contribution by nontypable Haemophilus influenzae in protracted and recurrent acute otitis media. Pediatr Infect Dis J 2009, 28(6):466-471.
• [6]Faden H, Duffy L, Wasielewski R, Wolf J, Krystofik D, Tung Y: Relationship between nasopharyngeal colonization and the development of otitis media in children: Tonawanda/Williamsville Pediatrics. J Infect Dis 1997, 175(6):1440-1445.
• [7]Cohen R, Levy C, Bingen E, Bechet S, Derkx V, Werner A, Koskas M, Varon E: [Nasopharyngeal carriage of children 6 to 60 months during the implementation of the 13-valent pneumococcal conjugate vaccine]. Arch Pediatr 2012, 19(10):1132-1139.
• [8]Kaur R, Chang A, Xu Q, Casey JR, Pichichero M: Phylogenetic relatedness and diversity of non-typable Haemophilus influenzae in the nasopharynx and middle ear fluid of children with acute otitis media. J Med Microbiol 2011, 60(Pt 12):1841-1848.
• [9]Murphy TF, Kirkham C: Biofilm formation by nontypeable Haemophilus influenzae: strain variability, outer membrane antigen expression and role of pili. BMC Microbiol 2002, 2:7.
• [10]Hall-Stoodley L, Hu FZ, Gieseke A, Nistico L, Nguyen D, Hayes J, Forbes M, Greenberg DP, Dice B, Burrows A, Wackym PA, Stoodley P, Post JC, Ehrlich GD, Kerschner JE: Direct detection of bacterial biofilms on the middle-ear mucosa of children with chronic otitis media. JAMA 2006, 296(2):202-211.
• [11]Costerton JW, Stewart PS, Greenberg EP: Bacterial biofilms: a common cause of persistent infections. Science 1999, 284(5418):1318-1322.
• [12]Otto M: Staphylococcal Infections: mechanisms of biofilm maturation and detachment as critical determinants of pathogenicity. Annu Rev Med 2013, 64:175-188.
• [13]Fernández-Olmos A, García-Castillo M, Maiz L, Lamas A, Baquero F, Cantón R: In vitro prevention of Pseudomonas aeruginosa early biofilm formation with antibiotics used in cystic fibrosis patients. Int J Antimicrob Agents 2012, 40(2):173-176.
• [14]Hotomi M, Yamanaka N, Shimada J, Ikeda Y, Faden H: Factors associated with clinical outcomes in acute otitis media. Ann Otol Rhinol Laryngol 2004, 113(10):846-852.
• [15]Moxon ER, Sweetman WA, Deadman ME, Ferguson DJP, Hood DW: Haemophilus influenzae biofilms: hypothesis or fact? Trends Microbiol 2008, 16(3):95-100.
• [16]Agence Française de sécurité sanitaire des produits de santé: Antibiotic therapy in current practice in upper respiratory tract infections in adults and children. arguments. Méd mal infect 2005, 35:635-694.
• [17]CASFM: Recommendations. 2012. Available at: http://www.sfm-microbiologie.org webcite
• [18]O'Toole GA, Kolter R: Initiation of biofilm formation in Pseudomonas fluorescens WCS365 proceeds via multiple, convergent signalling pathways: a genetic analysis. Mol Microbiol 1998, 28(3):449-461.
• [19]Sutherland I: Biofilm exopolysaccharides: a strong and sticky framework. Microbiology 2001, 147:3-9.
• [20]Spinali S: Pasteurellaceae Identification by MALDI-TOF Mass Spectrometry. MD Thesis. 2011.
• [21]Kadurugamuwa JL, Sin L, Albert E, Yu J, Francis K, DeBoer M, Rubin M, Bellinger-Kawahara C, Parr Jr TR Jr, Contag PR: Direct continuous method for monitoring biofilm infection in a mouse model. Infect Immun 2003, 71(2):882-890.
• [22]Soto SM, Smithson A, Horcajada JP, Martinez JA, Mensa JP, Vila J: Implication of biofilm formation in the persistence of urinary tract infection caused by uropathogenic Escherichia coli. Clin Microbiol Infect 2006, 12(10):1034-1036.
• [23]Niu C, Gilbert ES: Colorimetric method for identifying plant essential oil components that affect biofilm formation and structure. Appl Environ Microbiol 2004, 70(12):6951-6956.
• [24]Naves P, del Prado G, Huelves L, Gracia M, Ruiz V, Blanco J, Rodríguez-Cerrato V, Ponte MC, Soriano F: Measurement of biofilm formation by clinical isolates of Escherichia coli is method-dependent. J Appl Microbiol 2008, 105(2):585-590.
• [25]Norskov-Lauritsen N, Overballe MD, Kilian M: Delineation of the species Haemophilus influenzae by phenotype, multilocus sequence phylogeny, and detection of marker genes. J Bacteriol 2009, 191(3):822-831.
• [26]Bingen E, Cohen R, Jourenkova N, Gehanno P: Epidemiologic study of conjunctivitis-otitis syndrome. Pediatr Infect Dis J 2005, 24(8):731-732.
• [27]van Dongen TM, van der Heijden GJ, van Zon A, Bogaert D, Sanders EA, Schilder AG: Evaluation of concordance between the microorganisms detected in the nasopharynx and middle ear of children with otitis media. Pediatr Infect Dis J 2013, 32(5):549-552.
• [28]Swords WE: Nontypeable Haemophilus influenzae biofilms: role in chronic airway infections. Front Cell Infect Microbiol 2012, 2:97.
• [29]Langereis JD, Hermans PW: Novel concepts in nontypeable Haemophilus influenzae biofilm formation. FEMS Microbiol Lett 2013, 346(2):81-89.
• [30]Torretta S, Marchisio P, Drago L, Baggi E, De Vecchi E, Garavello W, Nazzari E, Pignataro L, Esposito S: Nasopharyngeal biofilm-producing otopathogens in children with nonsevere recurrent acute otitis media. Otolaryngol Head Neck Surg 2012, 146(6):991-996.
• [31]Webster P, Wu S, Gomez G, Apicella M, Plaut AG, St Geme JW III: Distribution of bacterial proteins in biofilms formed by non-typeable Haemophilus influenzae. J Histochem Cytochem 2006, 54(7):829-842.
• [32]Chavant P, Gaillard-Martinie B, Talon R, Hébraud M, Bernardi T: A new device for rapid evaluation of biofilm formation potential by bacteria. J Microbiol Methods 2007, 68(3):605-612.
• [33]Moriyama S, Hotomi M, Shimada J, Billal DS, Fujihara K, Yamanaka N: Formation of biofilm by Haemophilus influenzae isolated from pediatric intractable otitis media. Auris Nasus Larynx 2009, 36(5):525-531.
• [34]Rayner MG, Zhang Y, Gorry MC, Chen Y, Post JC, Ehrlich GD: Evidence of bacterial metabolic activity in culture-negative otitis media with effusion. JAMA 1998, 279(4):296-299.
• [35]Weimer KE, Armbruster CE, Juneau RA, Hong W, Pang B, Swords WE: Coinfection with Haemophilus influenzae promotes pneumococcal biofilm formation during experimental otitis media and impedes the progression of pneumococcal disease. J Infect Dis 2010, 202(7):1068-1075.
• [36]Tikhomirova A, Kidd SP: Haemophilus influenzae and Streptococcus pneumoniae: living together in a biofilm. Pathog Dis 2013, 69(2):114-126.
• [37]Slinger R, Chan F, Ferris W, Yeung SW, St Denis M, Gaboury I, Aaron SD: Multiple combination antibiotic susceptibility testing of nontypeable Haemophilus influenzae biofilms. Diagn Microbiol Infect Dis 2006, 56(3):247-253.
• [38]Starner TD, Zhang N, Kim G, Apicella MA, McCray PB Jr: Haemophilus influenzae forms biofilms on airway epithelia: im plications in cystic fibrosis. Am J Respir Crit Care Med 2006, 174(2):213-220.
• [39]Ehrlich GD, Veeh R, Wang X, Costerton JW, Hayes JD, Hu FZ, Daigle BJ, Ehrlich MD, Post JC: Mucosal biofilm formation on middle-ear mucosa in the chinchilla model of otitis media. JAMA 2002, 287(13):1710-175.
• [40]Sanchez CJ, Kumar N, Lizcano A, Shivshankar P, Dunning Hotopp JC, Jorgensen JH, Tettelin H, Orihuela CJ: Streptococcus pneumoniae in biofilms are unable to cause invasive disease due to altered virulence determinant production. PLoS One 2011, 6(12):e28738.