Current Issue
Online First
Volumes and Issues
For Authors
Manuscript Guidelines
Review Process
Conflict of Interest
Copyright
About the Journal
Editorial Board
Aim and Scope
Policy and Ethical Guidelines
Promotion of the Journal
Print-Version Subscription
Publisher and Contact Information
Contact Information
Sign in as an AUTHOR/REVIEWER
ORIGINAL ARTICLE
WIDEBAND ACOUSTIC IMMITTANCE (WAI) AND HISTOPATHOLOGY IN A MOUSE MODEL OF OTITIS MEDIA
1
Audiology Program, Communication Sciences and Disorders, Missouri State University, Springfield, United States
2
Communicative Disorders and Sciences, University at Buffalo, United States
3
Department of Otolaryngology, Washington University School of Medicine, Washington, United States
These authors had equal contribution to this work
A - Research concept and design; B - Collection and/or assembly of data; C - Data analysis and interpretation; D - Writing the article; E - Critical revision of the article; F - Final approval of article;
Submission date: 2025-05-23
Final revision date: 2025-11-08
Acceptance date: 2025-11-17
Online publication date: 2026-02-02
Publication date: 2026-02-02
Corresponding author
Wafaa Kaf
Audiology Program, Communication Sciences and Disorders, Missouri State University, 901 S National Avenue, 65897, Springfield, United States
Audiology Program, Communication Sciences and Disorders, Missouri State University, 901 S National Avenue, 65897, Springfield, United States
J Hear Sci 2025;15(4):30-45
KEYWORDS
TOPICS
ABSTRACT
Introduction:
This study aimed to: (1) quantify and compare middle ear status in two mice strains, C57BL/6J and CBA/CaJ, using wideband acoustic immittance (WAI) to measure wideband absorbance at ambient pressure (WBA) and at tympanometric peak pressures (WBT); (2) determine the percentage of mice with histologic evidence of otitis media (OM) after nasal inoculations with Bordetella hinzii; (3) assess how B. hinzii affects WBA and WBT; and (4) evaluate if antibiotic treatment reduces OM and restores absorbance to normal.
Material and methods:
Eight C57BL/6J and eight CBA/CaJ mice were used in Experiment 1. WBA and WBT (averaged across 0.5–8 kHz) were measured at baseline and 3 and 6 weeks after B. hinzii inoculation. Middle ear histopathology was performed to confirm the presence of OM. In Experiment 2, ten C57BL/6J mice with OM received antibiotics, and absorbance was tracked at baseline, during OM, and post-treatment.
Results:
(1) Baseline absorbance responses were reliably measured in both strains and both showed similar results with peak WBA (~0.4) near 1 kHz and maximal WBT at –50 daPa near 6–8 kHz; (2) None of the CBA/CaJ developed OM, whereas 13 of 16 C57BL/6J ears showed OM histologically; (3) WBA and WBT remained normal in CBA/CaJ mice post-inoculation. In C57BL/6J mice, WBA at ambient pressure was insensitive to OM, but WBT was significantly reduced at 3 and 6 weeks post-inoculation (p = 0.001); (4) Antibiotic-treated C57BL/6J mice showed WBT recovery as OM resolved histologically.
Conclusions:
Wideband acoustic immittance provides reliable absorbance measures in mice. C57BL/6J mice are susceptible to OM induced by B. hinzii, whereas CBA/CaJ are resistant. WBT can be used to detect and monitor OM in mice. Limitations of the study include a modest sample size and relative rather than absolute values of WBA and WBT due to species differences in calibration.
This study aimed to: (1) quantify and compare middle ear status in two mice strains, C57BL/6J and CBA/CaJ, using wideband acoustic immittance (WAI) to measure wideband absorbance at ambient pressure (WBA) and at tympanometric peak pressures (WBT); (2) determine the percentage of mice with histologic evidence of otitis media (OM) after nasal inoculations with Bordetella hinzii; (3) assess how B. hinzii affects WBA and WBT; and (4) evaluate if antibiotic treatment reduces OM and restores absorbance to normal.
Material and methods:
Eight C57BL/6J and eight CBA/CaJ mice were used in Experiment 1. WBA and WBT (averaged across 0.5–8 kHz) were measured at baseline and 3 and 6 weeks after B. hinzii inoculation. Middle ear histopathology was performed to confirm the presence of OM. In Experiment 2, ten C57BL/6J mice with OM received antibiotics, and absorbance was tracked at baseline, during OM, and post-treatment.
Results:
(1) Baseline absorbance responses were reliably measured in both strains and both showed similar results with peak WBA (~0.4) near 1 kHz and maximal WBT at –50 daPa near 6–8 kHz; (2) None of the CBA/CaJ developed OM, whereas 13 of 16 C57BL/6J ears showed OM histologically; (3) WBA and WBT remained normal in CBA/CaJ mice post-inoculation. In C57BL/6J mice, WBA at ambient pressure was insensitive to OM, but WBT was significantly reduced at 3 and 6 weeks post-inoculation (p = 0.001); (4) Antibiotic-treated C57BL/6J mice showed WBT recovery as OM resolved histologically.
Conclusions:
Wideband acoustic immittance provides reliable absorbance measures in mice. C57BL/6J mice are susceptible to OM induced by B. hinzii, whereas CBA/CaJ are resistant. WBT can be used to detect and monitor OM in mice. Limitations of the study include a modest sample size and relative rather than absolute values of WBA and WBT due to species differences in calibration.
ACKNOWLEDGEMENTS
The authors thank Suellen Greco for preparation of B. hinzii inoculates, Kelly G. Lytte for data collection, and Pat Keller for histology. We also thank the RStat Institute at Missouri State University for helping with data analysis.
FUNDING
This research was supported by the National Institute on Deafness and Other Communication Disorders (NIDCD) grants T35DC008765 (PI, William W. Clark) and P30DC004665 (PI, Richard A. Chole) and funding from the Graduate College of Missouri State University.
REFERENCES (59)
1.
Bhutta MF, Hedge EA, Parker A, Cheeseman MT, Brown SD. Oto-endoscopy: a reliable and validated technique for phenotyping otitis media in the mouse. Hear Res, 2011; 272(1–2): 5–12. https://doi.org/10.1016/j.hear....
2.
MacArthur CJ, Pillers DA, Pang J, Kempton JB, Trune DR. Altered expression of middle and inner ear cytokines in mouse otitis media. Laryngoscope, 2011; 121(2): 365–71. https://doi.org/10.1002/lary.2....
3.
Leichtle A, Lai Y, Wollenberg B, Wasserman SI, Ryan AF. Innate signaling in otitis media: pathogenesis and recovery. Curr Allergy Asthma Rep, 2011; 11(1): 78–84. https://doi.org/10.1007/s11882....
4.
Ryan AF, Ebmeyer J, Furukawa M, Pak K, Melhus A, Wasserman SI, Chung WH. Mouse models of induced otitis media. Brain Res, 2006; 1091(1): 3–8. https://doi.org/10.1016/j.brai....
5.
Lanphear BP, Byrd RS, Auinger P, Hall CB. Increasing prevalence of recurrent otitis media among children in the United States. Pediatrics, 1997; 99(3): E1. https://doi.org/10.1542/peds.9....
6.
Lim DJ, Chun YM, Lee HY, Moon SK, Chang KH, Li JD, Andalibi A. Cell biology of tubotympanum in relation to pathogenesis of otitis media: a review. Vaccine, 2000; 19 (Suppl. 1): S17–25. https://doi.org/10.1016/s0264-....
7.
Bodet E, Romeu C, Gomez X, Manonellas A, Palomar V. [Experimental model of recurrent acute otitis media: morphological study of the cochlear damage using scanning electron microscopy]. Acta Otorrinolaringol Esp, 2005; 56(4): 139–42. https://doi.org/10.1016/s0001-....
8.
Clark JM, Brinson G, Newman MK, Jewett BS, Sartor BR, Prazma J, Pillsbury HC. An animal model for the study of genetic predisposition in the pathogenesis of middle ear inflammation. Laryngoscope, 2000; 110(9): 1511–5. https://doi.org/10.1097/000055....
9.
Dewan KK, Taylor-Mulneix DL, Campos LL, Skarlupka AL, Wagner SM, Ryman VE, et al. A model of chronic, transmissible otitis media in mice. PLoS Pathog, 2019; 15(4): e1007696. https://doi.org/10.1371/journa....
10.
Giebink GS. Otitis media: the chinchilla model. Microb Drug Resist, 1999; 5(1): 57–72. https://doi.org/10.1089/mdr.19....
11.
Guan X, Seale TW, Gan RZ. Factors affecting sound energy absorbance in acute otitis media model of chinchilla. Hear Res, 2017; 350: 22–31. https://doi.org/10.1016/j.hear....
12.
Kariya S, Okano M, Fukushima K, Nomiya S, Kataoka Y, Nomiya R, Akagi H, Nishizaki K. Expression of inflammatory mediators in the otitis media induced by Helicobacter pylori antigen in mice. Clin Exp Immunol, 2008; 154(1): 134–40. https://doi.org/10.1111/j.1365....
13.
MacArthur CJ, Trune DR. Mouse models of otitis media. Curr Opin Otolaryngol Head Neck Surg, 2006; 14(5): 341–6. https://doi.org/10.1097/01.moo....
14.
Melhus A, Ryan AF. A mouse model for acute otitis media. APMIS, 2003; 111(10): 989–94. https://doi.org/10.1034/j.1600....
15.
Mitchell CR, Kempton JB, Scott-Tyler B, Trune DR. Otitis media incidence and impact on the auditory brain stem response in lipopolysaccharide-nonresponsive C3H/HeJ mice. Otolaryngol Head Neck Surg, 1997; 117(5): 459–64. https://doi.org/10.1016/S0194-....
16.
Zheng QY, Tong YC, Alagramam KN, Yu H. Tympanometry assessment of 61 inbred strains of mice. Hear Res, 2007; 231(1–2): 23–31. https://doi.org/10.1016/j.hear....
17.
Chen J, Ingham N, Clare S, Raisen C, Vancollie VE, Ismail O, et al. Mcph1-deficient mice reveal a role for MCPH1 in otitis media. PLoS One, 2013; 8(3): e58156. https://doi.org/10.1371/journa....
18.
Rosowski JJ, Brinsko KM, Tempel BI, Kujawa SG. The aging of the middle ear in 129S6/SvEvTac and CBA/CaJ mice: measurements of umbo velocity, hearing function, and the incidence of pathology. J Assoc Res Otolaryngol, 2003; 4(3): 371–83. https://doi.org/10.1007/s10162....
19.
Rye MS, Bhutta MF, Cheeseman MT, Burgner D, Blackwell JM, Brown SD, et al. Unraveling the genetics of otitis media: from mouse to human and back again. Mamm Genome, 2011; 22(1–2): 66–82. https://doi.org/10.1007/s00335....
20.
Trune DR, Zheng QY. Mouse models for human otitis media. Brain Res, 2009; 1277: 90–103. https://doi.org/10.1016/j.brai....
21.
Zheng QY, Johnson KR, Erway LC. Assessment of hearing in 80 inbred strains of mice by ABR threshold analyses. Hear Res, 1999; 130(1–2): 94–107. https://doi.org/10.1016/s0378-....
22.
Zheng T, Huang W, Yu H, Hu BH, Song P, McCarty CM, et al. gom1 mutant mice as a model of otitis media. J Assoc Res Otolaryngol, 2022; 23(2): 213–23. https://doi.org/10.1007/s10162....
23.
Margolis RH, Paul S, Saly GL, Schachern PA, Keefe DH. Wideband reflectance tympanometry in chinchillas and human. J Acoust Soc Am, 2001; 110(3 Pt 1): 1453–64. https://doi.org/10.1121/1.1394....
24.
Peake WT, Rosowski JJ. Impedance matching, optimum velocity, and ideal middle ears. Hear Res, 1991; 53(1): 1–6. https://doi.org/10.1016/0378-5....
25.
Voss SE, Rosowski JJ, Merchant SN, Peake WT. How do tympanic-membrane perforations affect human middle-ear sound transmission? Acta Otolaryngol, 2001; 121(2): 169–73. https://doi.org/10.1080/000164....
26.
AlMakadma H, Kei J, Yeager D, Feeney MP. Fundamental concepts for assessment and interpretation of wideband acoustic immittance measurements. Semin Hear, 2023; 44(1): 17–28. https://doi.org/10.1055/s-0043....
27.
Keefe DH, Sanford CA, Ellison JC, Fitzpatrick DF, Gorga MP. Wideband aural acoustic absorbance predicts conductive hearing loss in children. Int J Audiol, 2012; 51(12): 880–91. https://doi.org/10.3109/149920....
28.
Karuppannan A, Barman A. Wideband absorbance pattern in adults with otosclerosis and ossicular chain discontinuity. Auris Nasus Larynx, 2021; 48(4): 583–9. https://doi.org/10.1016/j.anl.....
29.
Karuppannan A, Barman A. Wideband absorbance tympanometry: a novel method in identifying otosclerosis. Eur Arch Otorhinolaryngol, 2021; 278(11): 4305–14. https://doi.org/10.1007/s00405....
30.
Karuppannan A, Barman A, Mamatha NM. Wideband absorbance pattern and its diagnostic value in adults with middle ear effusions and tympanic membrane perforation. J Int Adv Otol, 2024; 20(2): 158–63. https://doi.org/10.5152/iao.20....
31.
Feeney MP, Hunter LL, Kei J, Lilly DJ, Margolis RH, Nakajima HH, et al. Consensus statement: Eriksholm workshop on wideband absorbance measures of the middle ear. Ear Hear, 2013; 34 (Suppl 1): 78S–79S. https://doi.org/10.1097/AUD.0b....
32.
Guan X, Jiang S, Seale TW, Hitt BM, Gan RZ. Morphological changes in the tympanic membrane associated with Haemophilus influenzae-induced acute otitis media in the chinchilla. Int J Pediatr Otorhinolaryngol, 2015; 79(9): 1462–71. https://doi.org/10.1016/j.ijpo....
33.
Chen GD, Li L, McCall A, Ding D, Xing Z, Yu YE, Salvi R. Hearing impairment in murine model of Down syndrome. Front Genet, 2022; 13: 936128. https://doi.org/10.3389/fgene.....
34.
Leiberman A, Leibovitz E, Piglansky L, Raiz S, Press J, Yagupsky P, et al. Bacteriologic and clinical efficacy of trimethoprim–sulfamethoxazole for treatment of acute otitis media. Pediatr Infect Dis J, 2001; 20(3): 260–4. https://doi.org/10.1097/000064....
35.
Sanford CA, Keefe DH, Liu YW, Fitzpatrick D, McCreery RW, Lewis DE, et al. Sound-conduction effects on distortion-product otoacoustic emission screening outcomes in newborn infants: test performance of wideband acoustic transfer functions and 1-kHz tympanometry. Ear Hear, 2009; 30(6): 635–52. https://doi.org/10.1097/AUD.0b....
36.
Rosowski JJ, Mehta RP, Merchant SN. Diagnostic utility of laser-Doppler vibrometry in conductive hearing loss with normal tympanic membrane. Otol Neurotol, 2003; 24(2): 165–75. https://doi.org/10.1097/001294....
37.
Koay G, Heffner R, Heffner H. Behavioral audiograms of homozygous med(J) mutant mice with sodium channel deficiency and unaffected controls. Hear Res, 2002; 171(1–2): 111–8. https://doi.org/10.1016/s0378-....
38.
Radziwon KE, June KM, Stolzberg DJ, Xu-Friedman MA, Salvi RJ, Dent ML. Behaviorally measured audiograms and gap detection thresholds in CBA/CaJ mice. J Comp Physiol A Neuroethol Sens Neural Behav Physiol, 2009; 195(10): 961–9. https://doi.org/10.1007/s00359....
39.
Clark WW, Clark CS, Moody DB, Stebbins WC. Noise-induced hearing loss in the chinchilla, as determined by a positive-reinforcement technique. J Acoust Soc Am, 1974; 56(4): 1202–9. https://doi.org/10.1121/1.1903....
40.
Salvi RJ, Hamernik RP, Henderson D. Auditory nerve activity and cochlear morphology after noise exposure. Arch Otorhinolaryngol, 1979; 224(1–2): 111–6. https://doi.org/10.1007/BF0045....
41.
Vrettakos PA, Dear SP, Saunders C. Middle ear structure in the chinchilla: a quantitative study. Am J Otolaryngol, 1988; 9(2): 58–67. https://doi.org/10.1016/s0196-....
42.
Aithal S, Aithal V, Kei J, Anderson S, Liebenberg S. Eustachian tube dysfunction and wideband absorbance measurements at tympanometric peak Pressure and 0 daPa. J Am Acad Audiol, 2019; 30(9): 781–91. https://doi.org/10.3766/jaaa.1....
43.
Pan JL, Yang J. [The clinical value of wideband tympanometry in the diagnosis of otitis media with effusion]. Lin Chuang Er Bi Yan Hou Tou Jing Wai Ke Za Zhi, 2018; 32(17): 1309–15. https://doi.org/10.13201/j.iss... [in Chinese].
44.
Ahlstedt S. Experimental Escherichia coli 06 infection in mice. 1. Effect of immunisation on resistance in relation to 06 antibody levels in mice of different strains. Acta Pathol Microbiol Scand C, 1980; 88(1): 39–45.
45.
Rowe WP, Hartley JW. Genes affecting mink cell focus-inducing (MCF) murine leukemia virus infection and spontaneous lymphoma in AKR F1 hybrids. J Exp Med, 1983; 158(2): 353–64. https://doi.org/10.1084/jem.15....
46.
Dewan KK, Sedney C, Caulfield AD, Su Y, Ma L, Blas-Machado U, et al. Probing immune-mediated clearance of acute middle ear infection in mice. Front Cell Infect Microbiol, 2021; 11: 815627. https://doi.org/10.3389/fcimb.....
47.
Perniss A, Schmidt N, Gurtner C, Dietert K, Schwengers O, Weigel M, et al. Bordetella pseudohinzii targets cilia and impairs tracheal cilia-driven transport in naturally acquired infection in mice. Sci Rep, 2018; 8: 5681. https://doi.org/10.1038/s41598....
48.
Gautam A, Dixit S, Embers M, Gautam R, Philipp MT, Singh SR, et al. Different patterns of expression and of IL-10 modulation of inflammatory mediators from macrophages of Lyme disease-resistant and -susceptible mice. PLoS One, 2012; 7(9): e43860. https://doi.org/10.1371/journa....
49.
Zhao T, Ma P, Zhao F, Zheng T, Yan B, Zhang Q, et al. Phenotypic differences in the inner ears of CBA/CaJ and C57BL/6J mice carrying missense and single base pair deletion mutations in the Cdh23 gene. J Neurosci Res, 2021; 99(10): 2743–58. https://doi.org/10.1002/jnr.24....
50.
Mateo M, Generous A, Sinn PL, Cattaneo R. Connections matter: how viruses use cell–cell adhesion components. J Cell Sci, 2015; 128(3): 431–9. https://doi.org/10.1242/jcs.15....
51.
Ikeda K, Takasaka T. In vitro activity of ototopical drops against middle ear pathogens. Am J Otol, 1993; 14(2): 170–1.
52.
Tong MC, Woo JK, van Hasselt CA. A double-blind comparative study of ofloxacin otic drops versus neomycin–polymyxin B–hydrocortisone otic drops in the medical treatment of chronic suppurative otitis media. J Laryngol Otol, 1996; 110(4): 309–14. https://doi.org/10.1017/s00222....
53.
Kristjansson S, Skuladottir HE, Sturludottir M, Wennergren G. Increased prevalence of otitis media following respiratory syncytial virus infection. Acta Paediatr, 2010; 99(6): 867–70. https://doi.org/10.1111/j.1651....
54.
Post JC. Direct evidence of bacterial biofilms in otitis media. Laryngoscope, 2001; 111(12): 2083–94. https://doi.org/10.1097/000055....
55.
Quaranta A, Bartoli R, Resta L, Lozupone E. Candida and stapedial otosclerosis: histopathological findings. ORL J Otorhinolaryngol Relat Spec, 1992; 54(6): 334–6. https://doi.org/10.1159/000276....
56.
Thornton JL, Chevallier KM, Koka K, Gabbard SA, Tollin DJ. Conductive hearing loss induced by experimental middle-ear effusion in a chinchilla model reveals impaired tympanic membrane-coupled ossicular chain movement. J Assoc Res Otolaryngol, 2013; 14(4): 451–64. https://doi.org/10.1007/s10162....
57.
Norgaard KM, Fernandez-Grande E, Schmuck C, Laugesen S. Reproducing ear-canal reflectance using two measurement techniques in adult ears. J Acoust Soc Am, 2020; 147(4): 2334. https://doi.org/10.1121/10.000....
58.
Keefe DH, Bulen JC, Arehart KH, Burns EM. Ear-canal impedance and reflection coefficient in human infants and adults. J Acoust Soc Am, 1993; 94(5): 2617–38. https://doi.org/10.1121/1.4073....
59.
Aithal S, Kei J, Aithal V, Manuel A, Myers J, Driscoll C, Khan A. Normative study of wideband acoustic immittance measures in newborn infants. J Speech Lang Hear Res, 2017; 60(5): 1417–26. https://doi.org/10.1044/2016_J....
We process personal data collected when visiting the website. The function of obtaining information about users and their behavior is carried out by voluntarily entered information in forms and saving cookies in end devices. Data, including cookies, are used to provide services, improve the user experience and to analyze the traffic in accordance with the Privacy policy. Data are also collected and processed by Google Analytics tool (more).
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
You can change cookies settings in your browser. Restricted use of cookies in the browser configuration may affect some functionalities of the website.
