Middle Ear Fluid Cultures Research Articles

A novel immuno-molecular strategy for the detection of Streptococcus pneumoniae serotypes in human cerebrospinal and middle ear fluids

Streptococcus pneumoniae is one of the most common microorganisms causing acute otitis media (AOM) in children. While bacterial culture of middle ear fluid (MEF) is the gold standard to detect the etiological organisms, several host and pathogen factors impact the survival of the organisms resulting in false negatives. To overcome this limitation, we have developed and validated an innovative multiplex immuno-molecular assay to screen and detect the S. pneumoniae 15-valent pneumococcal conjugate vaccine (PCV15; STs 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, 22F, 23F, and 33F) vaccine serotypes in MEF. This novel in vitro approach involves two-step testing. First, the MEF specimens were tested for highly conserved pneumococcal genes, autolysin, lytA, and pneumolysin, ply using direct PCR to identify pneumococcus positive specimens. The pneumococcus positive specimens were screened for the presence of vaccine serotype specific pneumococcal polysaccharides using a 15-plex Pneumococcal Antigen Detection (PAD) assay, with specific capture and detection monoclonal antibodies. Due to the lack of availability of MEF samples, cerebrospinal fluid (CSF) was used as the surrogate matrix for the development and validation of the PCR-PAD assays. The PCR and PAD assays were separately evaluated for sensitivity and specificity. Subsequently, the PCR-PAD assays were cross-validated with human MEF samples (n = 39) which were culture confirmed to contain relevant bacterial strains. The combined PCR-PAD assays demonstrated high rate of agreement 94.9% (95% CI; 82.7, 99.4%) with historical Quellung serotype data of these MEF samples. This novel PCR-PAD assay demonstrates the feasibility of combining molecular and immunological assays to screen and identify PCV15 pneumococcal vaccine serotypes in AOM clinical samples.

Increase in Haemophilus influenzae Detection in 13-Valent Pneumococcal Conjugate Vaccine Immunized Children With Acute Otitis Media.

NTHi was the predominant pathogen in ear cultures from severe acute otitis media (AOM) episodes in PCV-13 vaccinated children, more commonly in girls. NTHi-AOM episodes were associated with more myringotomies due to a higher treatment failure incidence. The low rate of β-lactamase NTHi isolates in middle ear fluid cultures from PCV-13 vaccinated children presenting with AOM strengthens to still use amoxicillin as the first-line antibiotics.

Two decades of otitis media in northern Israel: Changing trends in the offending bacteria and antibiotic susceptibility

ObjectiveBacteriology and antibiotic resistance trends changed considerably following introduction of the pneumococcal conjugate vaccines (PCV) 7 and 13, with differences between geographic regions. The objective of this study was to evaluate changes in acute otitis media (AOM) bacteriology and antibiotic susceptibility from the pre-vaccination period (2002–2008) to after the introduction of PCV13 (2010–2019) in northern Israel. MethodsData were collected from 3277 middle ear fluid (MEF) cultures and 4822 common AOM-generating pathogens of children aged <5 years with otitis media, taken during 2002–2019. Age of the child, bacteriology, and antibiotic resistance were compared between 2002 and 2008, the pre-vaccination period when no vaccination was available and 2010–2019 when PCV13 was introduced. ResultsThe mean age of the children in the pre-vaccination and the vaccination periods was 18.7 ± 13.7 and 15.7 ± 12.5 months, respectively (p < 0.001); the mean age of those with group A streptococcus (GAS) positive cultures was older, p < 0.001.The prevalence of Streptococcus pneumoniae (S. pneumoniae) decreased between those periods, from 47% to 25.8%, p < 0.001, Haemophilus influenzae (H. influenza) increased from 38.4% to 47.1%, p < 0.001, GAS increased from 12.9% to 23.8%, p < 0.001, and Moraxella catarrhalis (M. cat) increased but not statistically significant from 1.7% to 3.1%. The yearly number of positive MEF cultures decreased from 395.1 to 205.6, p < 0.001. The antibiotic sensitivity rate of almost all antibiotics increased between the two study periods. ConclusionThe most common MEF bacteria in northern Israel today is H. influenzae. Comparing the pre-vaccination to the vaccination period, the incidence of S. pneumonia-positive cultures decreased while GAS and H. influenza cultures increased. The age of children with positive cultures increased, and the antibiotic sensitivity rate increased.KeyThis research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

BACTERIAL MENINGITIS: A SERIOUS COMPLICATION OF ACUTE OTITIS MEDIA

TOPIC: Critical Care TYPE: Medical Student/Resident Case Reports INTRODUCTION: Otitis media can be complicated by bacterial meningitis that can have high mortality and morbidity. With the advent of antibiotics this complication is significantly rare but delay in getting treatment for Otitis media increases the risk of meningitis exceptionally. (1) We present a case of otitis media complicated by bacterial meningitis in a young adult patient. CASE PRESENTATION: A 36 years old male presented with complaints of right ear pain and discharge. On presentation he was febrile to 102 F. On examination he was sick in appearance with a tender right ear and purulent discharge. He was intubated for airway protection and altered mental status. Initial lab investigations showed a WBC count of 24.3 k/ul. Lactic acid and Procalcitonin were raised. CT Scan of head showed right mastoid effusion. Blood cultures were sent and a Lumbar puncture with an opening pressure of 55 cmH2O was done. Patient was started on vancomycin and ceftriaxone along with intravenous steroids. CSF analysis showed a WBC count of 11400 k/ul, predominantly neutrophils. CSF protein content was >750 mg/dl with no glucose in it. A tympanostomy tube was placed in the right middle ear for drainage and purulent discharge was sent for culture and sensitivity. MRI Scan of head showed restricted diffusion in subarachnoid spaces consistent with pus and ventriculitis in the setting of meningitis without any evidence of abscess. Right sided mastoid effusion was also present. Magnetic resonance venography was negative for venous thrombosis. Blood, middle ear fluid and CSF cultures grew streptococcus pneumoniae and staphylococcus epidermidis. Patient improved clinically after 72 hours on antibiotics and was extubated after that with normalization of inflammatory markers. Antibiotics were tailored as per sensitivity data and will require mastoidectomy as outpatient after completion of intravenous antibiotics. DISCUSSION: Most common organisms causing otitis media and its complications include streptococcus pneumoniae, group A streptococci, staphylococcus aureus, H. Influenzae and pseudomonas aeruginosa. (2) In some cases no bacterial organism is isolated. Due to the development of antibiotic therapies the complications of otitis media can be controlled. Delay in receiving antibiotic therapy can complicate the middle ear infection. If otitis media is complicated by middle ear effusion then tympanostomy tube can help in drainage of the purulent material preventing the seeding of other surrounding structures. It can extend to mastoid and further to brain tissue if not treated in a timely fashion resulting in meningitis and abscess formation as happened in our patient it resulted in meningitis. Appropriate antibiotics with surgical intervention for drainage if needed is the course of treatment. CONCLUSIONS: Acute otitis media should be treated in a timely fashion to avoid the complication of bacterial meningitis. REFERENCE #1: 1. Harmes KM, Blackwood RA, Burrows HL, Cooke JM, Harrison RV, Passamani PP. Otitis media: diagnosis and treatment. Am Fam Physician. 2013 Oct 1;88(7):435-40. Erratum in: Am Fam Physician. 2014 Mar 1;89(5):318. Dosage error in article text. PMID: 24134083. REFERENCE #2: 2. Leichtle A, Hoffmann TK, Wigand MC. Otitis media – Definition, Pathogenese, Klinik, Diagnose und Therapie [Otitis media: definition, pathogenesis, clinical presentation, diagnosis and therapy]. Laryngorhinootologie. 2018 Jul;97(7):497-508. German. doi: 10.1055/s-0044-101327. Epub 2018 Jul 9. Erratum in: Laryngorhinootologie. 2018 Jul;97(7):E2. PMID: 29986368 DISCLOSURES: No relevant relationships by Muhammad Ibrar Islam, source=Web Response No relevant relationships by Namra Mahmood, source=Web Response No relevant relationships by Junaid mir, source=Web Response

Effectiveness of the 7- and 13-Valent Pneumococcal Conjugate Vaccines Against Vaccine-Serotype Otitis Media.

Despite the demonstrated impact of pneumococcal vaccine (PCV) implementation on otitis media (OM), demonstration of real-life serotype-specific effectiveness of the 7-valent and 13-valent PCVs (PCV7 and PCV13) is lacking owing to the paucity of culture-positive cases. Furthermore, prelicensure PCV13 efficacy against OM was not studied. The study was conducted from October 2009 to July 2013. Case patients were children aged 5-35 months with OM (mostly complex OM [recurrent/nonresponsive, spontaneously draining, chronic with effusion]) from whom middle-ear fluid culture was obtained; controls were contemporary children with rotavirus-negative gastroenteritis in a prospective population-based rotavirus surveillance, from the same age group with similar ethnic distribution and geographic location. Vaccine effectiveness (VE) was estimated as 1 minus the odds ratio using unconditional logistic regression, adjusting for time since PCV implementation, age, and ethnicity. A total of 223 case patients and 1370 controls were studied. Serotypes 19F and 19A together caused 56.1% of all vaccine-type (VT) OM. VE of ≥2 PCV doses in children aged 5-35 months was demonstrated as follows: PCV7 against OM due to PCV7 serotypes, 57.2% (95% confidence interval, 6.0%-80.5%); PCV13 against OM due to PCV13 serotypes, 77.4% (53.3%-92.1%); PCV13 against OM due to the 6 additional non-PCV7 serotypes 67.4% (17.6%-87.1%); PCV13 against OM due to serotype 19F, 91.3% (1.4%-99.2%); and PCV13 against OM due to serotype 3, 85.2% (23.9%-98.4%). PCV7 and PCV13 VE against OM due to serotype 19A in children aged 12-35 months was 72.4% (95% confidence interval, 6.2%-91.9%) and 94.6% (33.9%-99.6%), respectively. PCV7 and PCV13 were effective against complex OM caused by the targeted serotypes.

Is Multidrug Resistance in Acute Otitis Media with Streptococcus pneumoniae Associated with a More Severe Disease?

Objectives: Streptococcus pneumoniae is the leading bacterial etiologic agent in acute otitis media (AOM), and it produces a more severe inflammatory response than other otopathogens. Additionally, the presence of multidrug-resistant (MDR) S. pneumoniae is an important issue in the management of AOM. The present pilot study aimed to ascertain whether MDR S. pneumoniae is associated with a higher inflammatory response and/or a more severe disease. Methods: This was a prospective, single-center study on nonpneumococcal conjugate vaccine-immunized pediatric patients with severe AOM. Demographic and clinical characteristics were recorded. Middle ear fluid was obtained and cultured for each patient; antibiotic-resistance profiling was tested for S. pneumoniae isolates. The C-reactive protein (CRP) level and complete blood count were determined. Patients with positive middle ear fluid culture for S. pneumoniae were divided into 2 groups according to antibiotic resistance profile: MDR and non-MDR. Results: MDR S. pneumoniae was identified in 15 (35.7%) of the 42 eligible patients. Children in this group had significantly higher CRP levels (72.23 ± 62.92 vs. 14.96 ± 15.57 mg/L, p < 0.001), higher absolute neutrophil count (8.46 ± 3.97 vs. 5.22 ± 4.5 × 10³/mm³, p = 0.004), higher percentage of neutrophils (52.85 ± 13.49% vs. 38.34 ± 16.16%, p = 0.004), and were more prone to develop acute mastoiditis (p = 0.01). Receiver operating characteristic analysis identified CRP as the best biomarker to discriminate between the 2 groups of patients (AUC = 0.891). Conclusion: MDR S. pneumoniae was associated with a more severe inflammatory response and a higher incidence of mastoiditis.

Substantial reduction of antibiotic-non-susceptible pneumococcal otitis media following PCV7/PCV13 sequential introduction.

In the pre-pneumococcal conjugated vaccines (PCVs) era, serotypes included in the 7/13-valent PCVs (PCV7/PCV13) caused most pneumococcal otitis media (OM) and antibiotic-non-susceptible pneumococcal OM (ANSP-OM) episodes. In southern Israel, sequential PCV7/PCV13 introduction resulted in >90% reduction of vaccine-serotype OM. We assessed the dynamics of ANSP-OM necessitating middle ear fluid culture following PCV7/PCV13 sequential introduction in young children. This was a prospective, population-based, active surveillance. All episodes in children <3 years old, during 2004-16, were included. Two subperiods were defined: (i) pre-PCV: 2004-08; and (ii) PCV13: 2014-16. ANSP was defined for the following antibiotics: penicillin (MIC ≥0.1 mg/L and ≥1.0 mg/L), macrolide, tetracycline, clindamycin, ceftriaxone, trimethoprim/sulfamethoxazole and chloramphenicol. MDR was defined as ANSP for ≥3 classes. Overall, 2270 pneumococcal OM episodes were identified. Annual overall pneumococcal, PCV13 and non-PCV13 serotype OM incidence declined by 86%, 97% and 33%, respectively, comparing pre-PCV with the PCV13 period. During 2004-08, 95% of ANSP was observed in vaccine serotypes. Incidence of penicillin (MIC ≥0.1 mg/L and ≥1.0 mg/L), macrolide, tetracycline, clindamycin, ceftriaxone and multidrug ANSP-OM declined by >90% in the PCV13 period. Rates of trimethoprim/sulfamethoxazole and chloramphenicol ANSP-OM declined by 85% and 79%, respectively. The proportions of ANSP of all pneumococcal isolates declined by ∼70% for penicillin, ceftriaxone and erythromycin; 53% for tetracycline; and 55% for MDR, versus no significant reductions observed for chloramphenicol, trimethoprim/sulfamethoxazole and clindamycin. PCV7/PCV13 sequential introduction resulted in rapid and substantial ANSP-OM reduction, in parallel with the near disappearance of PCV13-serotype OM and no increase in replacement disease.

Studying PCV impact on clinical presentation of otitis media helps to understand its pathogenesis

BackgroundComplex otitis media (OM) may present with intact tympanic membrane or spontaneous otorrhea. We compared dynamics of intact tympanic membrane and spontaneous otorrhea OM following 7- and 13-valent conjugated vaccines (PCV7, PCV13) implementation, since differences in dynamics may imply different underlying mechanisms. MethodsA prospective, population-based, active surveillance. Episodes with middle-ear fluid cultures in children < 3 years were included. Defined sub-periods were: pre-pneumococcal conjugated vaccines (PCV) (2004–2008); PCV7 (2009–2011); PCV13 (2014–2016). ResultsOf 7705 episodes, 57.2% had intact tympanic membrane, 16.8% spontaneous otorrhea, 26.0% unknown.In the pre-PCV period, the spontaneous otorrhea group was older and had higher proportions of factors associated with recurrence/chronicity.During the PCV7 period, spontaneous otorrhea and intact tympanic membrane episodes caused by PCV13 serotypes decreased significantly (43% and 51%, respectively) and those caused by non-PCV13 serotypes and culture-negative episodes increased significantly. However, rates increases were steeper in the spontaneous otorrhea group for both non-PCV13 serotypes (117% vs. 38%) and culture-negative (720% vs. 69%). In the spontaneous otorrhea group, nontypeable Haemophilus influenzae rates increased non-significantly by 10% and all-cause OM rates increased significantly by 56%, while in the intact tympanic membrane group the respective rates decreased significantly by 22% and 11%. These trends were especially pronounced in ages 24–35 months.Despite these differences, after PCV13 introduction, both spontaneous otorrhea and intact tympanic membrane rates declined for all outcomes. ConclusionsSpontaneous otorrhea was associated with older age, frequent history of complex OM and delayed PCV impact, suggesting a higher proportion of advanced-stage complex OM.

Pneumococcal Phenotype and Interaction with Nontypeable Haemophilus influenzae as Determinants of Otitis Media Progression.

All-cause otitis media (OM) incidence has declined in numerous settings following introduction of pneumococcal conjugate vaccines (PCVs) despite increases in carriage of nonvaccine pneumococcal serotypes escaping immune pressure. To understand the basis for the declining incidence, we assessed the intrinsic capacity of pneumococcal serotypes to cause OM independently and in polymicrobial infections involving nontypeable Haemophilus influenzae (NTHi) using samples obtained from middle ear fluid and nasopharyngeal cultures before PCV7/13 rollout. Data included samples from OM episodes (11,811) submitted for cultures during a 10-year prospective study in southern Israel and nasopharyngeal samples (1,588) from unvaccinated asymptomatic children in the same population. We compared data representing pneumococcal serotype diversity across carriage and disease isolates with and without NTHi coisolation. We also measured associations between the pneumococcal phenotype and the rate of progression from colonization to OM in the presence and absence of NTHi. Whereas pneumococcal serotype diversity was lower in single-species OM than in single-species colonization, levels of serotype diversity did not differ significantly between colonization and OM in mixed-species episodes. Serotypes differed roughly 100-fold in progression rates, and those differences were attenuated in polymicrobial episodes. Vaccine serotype pneumococci had higher rates of progression than nonvaccine serotypes. While serotype invasiveness was a weak predictor of the OM progression rate, efficient capsular metabolic properties-traditionally thought to serve as an advantage in colonization-predicted an enhanced rate of progression to complex OM. The lower capacity of nonvaccine serotypes to cause OM may partially account for reductions in all-cause OM incidence despite serotype replacement in carriage following rollout of PCVs.

Three Innate Cytokine Biomarkers Predict Presence of Acute Otitis Media and Relevant Otopathogens

1.1Diagnosis of Acute Otitis Media (AOM) is challenging, resulting in frequent over diagnosis and improper prescription of antibiotics. A serum biomarker of AOM would significantly improve pediatric care for this common illness. 1.2Serum samples were studied from 197 children 6-36 months old during health, during viral Upper Respiratory Infection (URI) without middle ear involvement, and at the onset of AOM (confirmed by tympanocentesis). Serum concentrations of S100A12, IL-10, and ICAM-1 were measured by ELISA. Otopathogens were identified by culture of middle ear fluid. A predictive model for infection and causative otopathogen was developed based on density distributions of the measured cytokines. 1.3A biomarker score derived from subject age and serum concentrations of S100A12, IL-10, and ICAM-1 was significantly able to distinguish both between health and disease and between upper respiratory infections with and without middle ear involvement (AOM vs URI), and further predicted the specific causative bacterial pathogen. This biomarker could also identify recurrent OM-prone children. 1.4For the first time we show that a biomarker risk score derived from serum cytokine levels can predict the presence of bacterial AOM, the likely Otopathogen, and the recurrent OM-prone child. 1.5(1) AOM is a widespread pediatric infection with a substantial economic burden. (2) Three serum cytokines can discriminate between URI and AOM, reducing over diagnosis. (3) Prediction of responsible pathogen enables targeted antibiotic prescription.

Haemophilus influenzae-protein D specific antibody correlate with protection against acute otitis media in young children

BackgroundHaemophilus influenzae (Hi) causes respiratory infections and pathogenesis of this microbe begins in the human nasopharynx (NP). The objective of this study was to assess the correlation of NP colonization-induced serum antibody levels to Hi protein D with risk of acute otitis media (AOM) in children <2 yr. Methods455 sera from 213 children (age 6–24 months old) were collected when they were colonized with Hi and when the children developed AOM. Presence of Hi during AOM was confirmed by culture of middle ear fluid. Quantitative ELISA was used to determine serum IgG against protein D antigen. ResultsAsymptomatic Hi NP colonization reduced the risk of future AOM infections. Higher serum IgG titers against Hi protein D were correlated with reduced future AOM risk. ConclusionColonization by Hi reduces future AOM risk. Higher antibody levels against protein D correlates with lower risk of AOM caused by Hi.

Epidemiology of Acute Otitis Media in the Postpneumococcal Conjugate Vaccine Era.

To study the epidemiology of acute otitis media (AOM), especially the otitis-prone condition, during the pneumococcal conjugate vaccines 7 and 13 era. Six hundred and fifteen children were prospectively managed from 6 to 36 months of life during a 10-year time frame (June 2006-June 2016). All clinical diagnoses of AOM were confirmed by tympanocentesis and bacterial culture of middle ear fluid. By 1 year of age, 23% of the children experienced ≥1 episode of AOM; by 3 years of age, 60% had ≥1 episodes of AOM, and 24% had ≥3 episodes. The peak incidence occurred at 6 to 12 months of life. Multivariable analysis of demographic and environmental data revealed a significantly increased risk of AOM associated with male sex, non-Hispanic white race, family history of recurrent AOM, day care attendance, and early occurrence of AOM. Risk factors for stringently defined (tympanocentesis-confirmed) otitis proneness, in which children suffered at least 3 episodes of AOM in a 6-month period or at least 4 within a year, were male sex, day care attendance, and family history of AOM, whereas breastfeeding in the first 6 months of life was protective. Stringently defined otitis prone children were also likely to experience their first AOM episode at a younger age. The proportion of Streptococcus pneumoniae, Haemophilus influenzae, and Moraxella catarrhalis causing AOM had dynamic changes during the past decade. We conclude that the epidemiology but not the risk factors for AOM have undergone substantial changes since the introduction of pneumococcal conjugate vaccines.

Pan-serotype Reduction in Progression of Streptococcus pneumoniae to Otitis Media After Rollout of Pneumococcal Conjugate Vaccines.

Reductions in otitis media (OM) burden following rollout of pneumococcal conjugate vaccines (PCVs) have exceeded predictions of vaccine impact. In settings with active surveillance, reductions in OM caused by vaccine-targeted pneumococcal serotypes have co-occurred with reductions in OM caused by other pathogens carried in the upper-respiratory tract of children. To understand these changes, we investigated the progression of vaccine-targeted and non-vaccine pneumococcal serotypes from carriage to OM before and after vaccine rollout. Nasopharyngeal carriage prevalence of pneumococcus was monitored in prospective studies of Bedouin and Jewish children <3 years old in southern Israel between 2004 and 2016. Incidence of OM necessitating middle-ear fluid culture (predominantly complex OM including recurrent, spontaneously-draining, non-responsive, and chronic cases) was monitored via prospective, population-based active surveillance. We estimated rates of pneumococcal serotype-specific progression from carriage to disease before and after rollout of PCV7/13, measured as OM incidence per carrier. We pooled serotype-specific estimates using Bayesian random-effects models. On average, rates of progression declined 92% (95% credible interval: 79-97%) and 80% (46-93%) for PCV7/13 serotypes among Bedouin and Jewish children <12 months old, respectively, and 32% (-58-71%) and 61% (-5-86%) among children aged 12-35m. For non-vaccine serotypes, rates of progression among Bedouin and Jewish children aged <12m declined 74% (55-85%) and 43% (4-68%), respectively. Vaccine-targeted and non-vaccine pneumococcal serotypes showed lower rates of progression to complex OM after rollout of PCV7/13. Early-life OM episodes historically associated with vaccine-serotype pneumococci may impact the susceptibility of children to OM progression.

Using Pneumococcal Carriage Data to Monitor Postvaccination Changes in the Incidence of Pneumococcal Otitis Media.

Pneumococcal conjugate vaccines (PCVs) have substantially reduced the burden of pneumococcal disease, including the incidence of otitis media (OM). However, in most countries, no surveillance exists to monitor the change in pneumococcal OM incidence after the introduction of PCVs. We explored whether measuring pneumococcal carriage was a useful surrogate for monitoring postvaccination changes in the incidence of pneumococcal OM. The 7-valent PCV was introduced to Israel's national immunization program in July 2009 and gradually replaced by the 13-valent PCV starting in November 2010. Each day since 2009, nasopharyngeal swabs have been obtained from the first 4 Bedouin children and the first 4 Jewish children who were younger than 5 years old and attended a pediatric emergency room in southern Israel. During the same time, OM surveillance in southern Israel included all children younger than 2 years of age who were diagnosed with OM and had undergone a middle-ear fluid culture. The relative change in the prevalence of vaccine-serotype (VT) pneumococcal carriage was predictive of the relative change in incidence of OM due to VT pneumococcus. However, the serotype replacement observed in non-VT carriage is not paralleled in the incidence of OM due to non-VT pneumococcus. This could indicate that there are more complex mechanisms of the immune response involved in preventing initial and consecutive episodes of OM, which has been changed through declining prevalence of the most virulent serotypes as a result of vaccination.

Ten-Year Study of the Stringently Defined Otitis-prone Child in Rochester, NY.

This review summarizes a prospective, longitudinal 10-year study in Rochester, NY, with virtually every clinically diagnosed acute otitis media (AOM) confirmed by bacterial culture of middle ear fluid. Children experiencing 3 episodes within 6 months or 4 episodes in 12 months were considered stringently defined otitis prone (sOP). We found stringent diagnosis compared with clinical diagnosis reduced the frequency of children meeting the OP definition from 27% to 6% resulting in 14.8% and 2.4% receiving tympanostomy tubes, respectively. Significantly more often respiratory syncytial virus infection led to AOM in sOP than non-otitis-prone children that correlated with diminished total respiratory syncytial virus-specific serum IgG. sOP children produced low levels of antibody to Streptococcus pneumoniae and Haemophilus influenzae candidate vaccine protein antigens and to routine pediatric vaccines. sOP children generated significantly fewer memory B cells, functional and memory T cells to otopathogens following nasopharyngeal colonization and AOM than non-otitis-prone children and they had defects in antigen-presenting cells.

Ten-Year Study of Acute Otitis Media in Rochester, NY.

This review summarizes a prospective, longitudinal 10-year study in Rochester, NY, involving 760 children where virtually all clinically diagnosed acute otitis media (AOM) was confirmed by bacterial culture of middle ear fluid. This review describes detection of otopathogens in middle ear fluid, nasopharyngeal (NP) otopathogen colonization patterns, AOM risk factor analysis, biomarkers of AOM and antibody responses to NP colonization by otopathogens. After licensure of PCV13, there was an immediate drop in AOM caused by Streptococcus pneumoniae (Spn) vaccine serotypes and shortly thereafer an increase in nonvaccine types 16, 21 and 35B. When NP co-colonization occurred, nontypeable Haemophilus influenzae (NTHi) predominated over Spn to cause AOM, and NTHi and Spn both predominated over Moraxella catarrhalis. Transcriptome analysis of peripheral blood mononuclear cells identified unique signatures for NTHi AOM compared with Spn AOM. Elevation of 3 cytokines in serum (S100A12, intercellular adhesion molecule 1 and interleukin 10) accurately predicted the presence and recovery from AOM and the likely otopathogen. NP colonization was an immunizing event.

Efficacy of Solithromycin (CEM-101) for Experimental Otitis Media Caused by Nontypeable Haemophilus influenzae and Streptococcus pneumoniae.

Solithromycin (CEM-101) is a "fourth-generation" macrolide, as it has three binding site and is acid stable. The three binding sites confer activity against bacteria resistant to the older macrolides and ketolides, including multidrug-resistant Streptococcus pneumoniae and nontypeable Haemophilus influenzae (NTHi). The objective of this study was to evaluate solithromycin pharmacokinetics (PK), middle ear fluid (MEF) concentrations, and microbiologic efficacy in a chinchilla model of experimental otitis media (EOM) due to strains of S. pneumoniae or NTHi. Plasma PK (maximum concentration of drug in serum [Cmax] and area under the concentration-time curve from 0 to 24 h [AUC0-24]) and middle ear fluid (MEF) concentrations were determined. Isolates with specified antimicrobial susceptibility patterns were inoculated directly into the middle ear (ME). Plasma and MEF were collected for PK and MEF cultures performed to determine efficacy. Solithromycin administered at 150 mg/kg of body weight/day resulted in Cmax and AUC0-24 values of 2.2 μg/ml and 27.4 μg · h/ml in plasma and 1.7 μg/ml and 28.2 μg · h/ml in extracellular MEF on day 1. By day 3, Cmax and AUC0-24 values had increased to 4.5 μg/ml and 54 μg · h/ml in plasma and 4.8 μg/ml and 98.6 μg · h/ml in extracellular MEF. For NTHi EOM, three isolates with MIC/minimal bactericidal concentration (MBC) ratios of 0.5/1 μg/ml (isolate BCH1), 2/2 μg/ml (isolate BMC1247C), and 4/4 μg/ml (isolate BMC1213C) were selected. The MEF of >85% of animals infected with BCH1 and BMC1247C was sterilized. For NTHi BMC1213, >85% of MEF cultures remained positive. For S. pneumoniae EOM, 3 isolates with MIC/MBC ratios of 0.06/0.125 μg/ml (S. pneumoniae 331), 0.125/1 μg/ml (S. pneumoniae CP-645 [MLSB phenotype]), and 0.5/2 μg/ml (CP-712 [mefA subclass mefA resistance]) were selected. Solithromycin sterilized MEF in 100% of animals infected with S. pneumoniae 331 and S. pneumoniae CP-645. ME infection persisted in 60% of animals infected with CP-712. In a model of EOM, solithromycin sterilized MEF in >85% of animals challenged with NTHi with an MIC of ≤2 μg/ml and 100% of ME infected with S. pneumoniae with an MIC of ≤0.125 μg/ml.

Impact of Widespread Introduction of Pneumococcal Conjugate Vaccines on Pneumococcal and Nonpneumococcal Otitis Media.

Pneumococcal conjugated vaccines (PCVs) impact on complex otitis media (OM; including recurrent, nonresponsive, and chronic OM with effusion) was greater than that on simple, acute OM in previous studies. Since complex OM is often a polymicrobial disease, we speculated that reduction of complex OM by PCVs would be associated with reduction of non-pneumococcal OM. In a prospective, population-based, active surveillance, all OM episodes submitted for middle ear fluid culture in children <3 years from 2004 through 2015 were included. Three sub-periods were established: pre-PCV, PCV7, and PCV13. Incidence rate ratios (IRRs) comparing the 3 periods were calculated for pneumococcal, nontypable Haemophilus influenzae (NTHi), Moraxella catarrhalis, Streptococcus pyogenes, and culture-negative OM. Overall, 7475 episodes were included. Of all-NTHi cases in the pre-PCV period, 34% were mixed with Streptococcus pneumoniae IRRs (95% confidence interval) comparing the pre-PCV to the PCV13 period were 0.02 (0.01-0.04), 0.12 (0.08-0.20), and 0.18 (0.15-0.21) for PCV7+6A serotypes, 5 additional PCV13 serotypes, and all-pneumococcal OM, respectively; non-PCV13 serotype episodes were not significantly reduced. IRRs for single NTHi, mixed NTHi + S. pneumoniae, and all-NTHi OM were 0.30 (0.25-0.35), 0.18 (0.13-0.24), and 0.25 (0.22-0.29), respectively. Moraxella catarrhalis, S. pyogenes, and culture-negative episodes were also significantly reduced. Both pneumococcal and non-pneumococcal OM episodes, enriched with complex cases, declined substantially in children <3 years following sequential PCV7/PCV13 introduction. The reduction in non-pneumococcal episodes may be attributed to early OM episodes prevention, resulting in a lower rate of complex, often non-pneumococcal OM.

Changing trends of acute otitis media bacteriology in central Israel in the pneumococcal conjugate vaccines era.

The widespread use of 7-valent pneumococcal conjugate vaccine (PCV7) has changed acute otitis media (AOM) bacteriology. Only scattered data with regard to this effect of PCV13 have been published so far. We retrospectively identified children <6 years of age who presented to our hospital with AOM, and had middle ear fluid (MEF) cultures obtained during tympanocentesis or from spontaneous otorrhea during 2008-2013, when PCV7 (2009) and PCV13 (2010) were gradually introduced in the Israeli National Immunization Program. Data were extracted for demographics, clinical and microbiologic parameters, according to vaccination status. Of the 295 eligible AOM episodes reported in 279 children, 224 (76%) had MEF cultures from tympanocentesis and 71 (24%) from spontaneous otorrhea. Boys and children <2 years of age contributed 178 (60%) and 219 (74%) AOM episodes, respectively. Acute mastoiditis complicated 58 (20%) of these episodes. None of the children were PCV immunized in 2008, but >90% had received ≥1 PCV dose(s) by 2011 or later. Of the 106 (36%) MEF cultures which tested positive for otopathogens, Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and multiple bacteria grew in 60 (57%), 39 (37%), 2 (2%) and 5 (5%) episodes, respectively. S. pneumonia-positive MEF culture rate in unimmunized children (31, 69%) was significantly higher than in PCV7-immunized children (22, 59%) or PCV13-immunized children (12, 50%), P = 0.04 and P = 0.02, respectively. PCV7 and PCV13 implementations in the Israeli National Immunization Program were associated with a rapid reduction of "severe" pneumococcal AOM episodes.

Could fungi be detected in the fluid of persistent otitis media with effusion?

Abstract Otitis media with effusion (OME) often is considered a direct extension of the inflammatory process that occurs during long-lasting or recurrent episodes of acute otitis media. The observations above suggest that OME has an infectious etiology. Most bacterial and viral cultures of middle ear fluid that had been performed were often negative suggesting that other infectious agents may be involved such as fungi. Thirty patients (group A) suffering from chronic secretory otitis media (OME) were enrolled in this study. Three samples were collected and investigated using PCR assay with universal fungal primers and Sabouraud agar. The first sample was obtained from the fluid of the middle ear before insertion of the ventilation tube; the second sample was obtained from nasal secretions; and the third sample was obtained from the ipsilateral peritubal area of the nasopharynx. Thirty patients (group B) with comparable age group without history of ear diseases scheduled for tonsillectomy or adenotonsillectomy were added as a control group. Samples from peritubal area of the nasopharynx of patients and nasal secretion were tested using PCR assay with universal fungal primers and Sabouraud agar. PCR examination of the middle ear aspirate in group (A) cases was positive in 7 cases (23.3%), in nasal secretions samples 2 cases only (13.3%) were positive and no positive cases were detected in nasopharyngeal swab samples. In group (A), Sabouraud agar culture was positive for fungal culture of middle ear aspirates in 5 cases (16.6%) but in no cases for nasal secretion samples. Group A showed also negative (N0) growth in 30 (100%) patients for nasopharyngeal swab on Sabouraud agar. In group B, the findings of nasopharyngeal swab were negative (N0) growth in all examined samples on Sabouraud agar, and nasal secretions were also negative for fungal DNA detection using PCR assay. In this study, fungal DNA could be detected in the middle ear fluid in seven (23.3%) of 30 patients with persistent OME using PCR assay, and fungi could be detected in five (16.6%) patients on Sabouraud agar. A significant relationship was found between detection of fungi in the middle ear fluid and the duration of the disease, associated adenoid, and history of asthma.