Livjot Sachdeva

Identification of adenoid biofilms with middle ear pathogens in otitis-prone children utilizing SEM and FISH

OBJECTIVES Biofilms have been implicated in the development of several chronic infections. We sought to demonstrate middle ear pathogens in adenoid biofilms using scanning electron microscopy (SEM) and fluorescent in situ hybridization (FISH) with confocal laser scanning microscopy (CLSM). METHODS Comparative micro-anatomic investigation of adenoid mucosa using SEM and FISH with confocal scanning laser microscopic (CLSM) imaging from patients with recurrent acute otitis media (RAOM). RESULTS All otitis-prone children demonstrated biofilm surface area presence greater than 85% by SEM. FISH accompanied by CLSM imaging also demonstrated patchy biofilms All biofilms contained middle ear pathogens and were frequent in polymicrobial distributions: 4 of 6, 4 of 6 and 3 of 6 samples contained Haemophilus influenzae, Streptococcus pneumoniae and Moraxella catarrhalis, respectively. CONCLUSIONS Dense adenoid biofilms may act as a reservoir for reinfection of the tubotympanum. Aspiration of planktonic middle ear pathogens existing in resistant adenoid biofilms during a viral upper respiratory tract infection may be an important event in the development of RAOM.

Demonstration of Nasopharyngeal and Middle Ear Mucosal Biofilms in an Animal Model of Acute Otitis Media

Objectives: We performed this study to determine the role of nasopharyngeal and middle ear (ME) biofilms in acute otitis media (AOM). Methods: Sixty female 6-month-old chinchillas, free of ME disease, were utilized. Experimental animals were inoculated with influenza A followed by Streptococcus pneumonia 7 days later. Control animals were inoculated with Sorensens phosphate buffer. Daily otoscopy and tympanometry was performed, and the animals were painlessly sacrificed on days 1, 2, 5, 8, and 14. All mucosae were harvested and prepared for scanning electron microscopy. Results: The ME inflammation, initially detected on day 2 after bacterial inoculation, peaked on day 8. Eight percent of the dually inoculated chinchillas displayed type B tympanograms, and 40% displayed type C. Otoscopic evaluation of tympanic membrane inflammation was rated from 0 to 4 (0 = normal and 4 = severe drainage and/or inflammation) according to an otoscopic grading system. Ten percent of the experimental chinchillas had a grade 2 score, 20% had grade 3, and 6.7% had grade 4. The controls demonstrated no abnormal tympanometric or otoscopic findings. Scanning electron microscopic imaging showed dense biofilms on 83% of the nasopharynges and 67% of the MEs on day 8 in the experimental animals. All animals with ME biofilms had biofilms in the nasopharynx. The controls did not demonstrate biofilm formation. Conclusions: The study parallels the natural pathogenesis of AOM in humans. The demonstration of mucosal biofilms in both the nasopharynx (58%) and the ME (47%) of animals with ME inflammation and/or infection lends further support to the importance of mucosal biofilms in the pathogenesis of AOM.

Biofilms and chronic otitis media: an initial exploration into the role of biofilms in the pathogenesis of chronic otitis media

PURPOSE The aim of the study was to compare the extent of biofilm infection in percentage of mucosal surface area of adenoids removed from children with otitis media with effusion (OME) vs those with recurrent acute otitis media (RAOM) and obstructive sleep apnea (OSA). MATERIALS AND METHODS Comparative microanatomical investigation of adenoid mucosa using scanning electron microscopy obtained from 30 children with OME, RAOM, and OSA was used in this study. Seventeen males and 13 females ranging in age from 9 months to 10 years were included in this study. Percentage of biofilm surface area involvement was the main measure. RESULTS Adenoids removed from patients with OME had moderately dense mature biofilms covering the mucosal surface with a mean of 27.7% of their mucosal surface covered with mature biofilms. These results were distinct from results obtained from patients diagnosed with RAOM and OSA with means of 97.6% and 0.10% of their mucosal surfaces covered with mature biofilms, respectively. These differences were statistically significant at P < .0001. CONCLUSIONS Adenoids removed from patients with OME were characterized by distinctly different percentage of biofilm mucosal surface area coverage, with significantly more biofilm presence than OSA patients but significantly less biofilm presence than RAOM patients. Although previous investigations have supported a dominant role of nasopharyngeal biofilms in RAOM pathogenesis, these results suggest nasopharyngeal biofilms may play a different role in the pathogenesis of OME and that this clinical entity may be more multifactorial in nature.

New paradigms in the pathogenesis of otitis media in children.

Acute otitis media (AOM) is a multifactorial disease with a significant socioeconomic impact. The pathogenesis of AOM is attributed to a variety of well-established internal and extrinsic factors. Recent evidence strongly points to bacterial biofilm formation as an important contributor to this disease entity. The nasopharynx is a likely reservoir for infection with subsequent seeding of pathogens to the middle ear via planktonic shedding. Various modalities have been used to directly detect biofilm formation in the middle ear mucosa of children with AOM. Further insights into this disease may lead to new strategies for prevention and treatment.

Potential role of biofilms in deep cervical abscess

OBJECTIVES Deep neck abscesses are complex head & neck problems that can lead to significant complications including life threatening infections. It is understood that the pathology of these infections is primarily polymicrobial. Although broad spectrum antibiotics can be effective for small abscesses, larger abscesses can be recalcitrant and difficult to treat with antibiotics. It has been demonstrated for several infectious diseases, including some of head & neck, that biofilm phenotypes present a unique model for recurrence and chronicity of infectious diseases. It is suspected that biofilm phenotypes could play a crucial role in the recalcitrance of large deep neck abscesses. This study presents initial evidence indicating the presence of polymicrobial biofilms in deep neck space infections. METHODS Fourteen samples obtained via biopsy of abscess walls from deep neck spaces of patients undergoing surgical drainage. Eight patients were male and 6 were female. All but one patient were pediatric with ages ranging from 18 months to 32 years. All samples were processed and analyzed with scanning electron microscopy. RESULTS Electron micrographs of 12 out of 14 specimens showed discrete biofilm architecture with individual bacteria, both rods and cocci, embedded within the matrix. This was starkly different from tissue surfaces devoid of biofilms. CONCLUSIONS This initial evidence suggests that biofilm phenotypes could play a role in the pathogenesis and recalcitrance of deep neck infections, particularly in larger abscesses.

Frequency of Otitis Media Based on Otoendoscopic Evaluation in Preterm Infants

Objective This study was conducted to determine the frequency of otitis media in preterm neonates using otoendoscopy and tympanometry. Study Design Prospective study. Setting Wayne State University, Hutzel Women’s Hospital Neonatal Intensive Care Unit. Subjects and Methods Eighty-six preterm infants were included (gestational age <36 weeks). Otoendoscopy and tympanometry were performed to detect the presence of otitis media. Kappa statistic and logistic regression were used for statistical analysis. Results Otoendoscopy was performed in 85 patients. The frequency of otoendoscopy-diagnosed otitis media was 72.9% (62/85). Tympanometry could be performed on 69.76% of the ears. There was 73.5% agreement between the findings of tympanometry and those of otoendoscopy. The association between the presence of otitis media and gestational age at birth was statistically significant. The lower the gestational age, the higher the frequency of otoendoscopy-diagnosed otitis media (P = .001). Conclusion Otoendoscopically diagnosed otitis media is frequent in preterm neonates. There was agreement between the results of tympanometry and those of otoendoscopy. The frequency of otitis media increased with lower gestational age.

Grand Challenges in Pediatric Otolaryngology

The field of pediatric otolaryngology has undergone significant changes over the past two decades. These changes have resulted in new areas of investigations and challenges. This article will summarize some of the more recent advances and challenges facing this field. Specific focus of this grand challenge will include: (1) Role of biofilms in infectious diseases in otolaryngology, (2) tissue engineering in pediatric airway reconstructive surgery, (3) minimally invasive surgical techniques, and (4) advances in molecular biology of sensorineural hearing loss (SNHL).

Incidence of Otitis Media in Preterm Infants

Objectives: 1) Determine if objective tympanometry findings correlate with the subjective endoscopy finding in diagnosing middle ear disease (AOM or OME) in premature infants. 2) Determine if the incidence of middle ear disease increases with lower gestational weight in premature infants. Methods: Our prospective study includes 76 preterm infants, (< 36wks) admitted to the Neonatal Intensive Care Unit (NICU) at Harper Hospital. Endoscopic otoscopy and tympanometry were performed to detect the presence of otitis media within the first 4 days of birth. The Fisher exact test was used for statistical analysis. Results: 1) 44 ears had normal otoendoscopic findings, of which 35 had normal tympanometry findings (79.54 %). 63 ears had middle ear disease, of which 48 showed abnormal reading on tympanometry (76.19 %). 2) 26 babies weighed less than 1300 grams at birth (average gestational age 26.99 weeks), 23 of which had middle ear disease on otoendoscopy. 25 babies weighed more than 1300 grams (average gestational age 31.85 weeks), 12 of which had middle ear disease on otoendoscopy. A P value of 0.0025 suggests a significant correlation between low birth weight and middle ear disease. Conclusions: We demonstrate that endoscopic otoscopy technique correlates with objective tympanometry data, although there are several limitations to comparing these techniques. We provide evidence that the incidence of otitis media in LBW preterm infants is higher based on otoendoscopy.