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Children diagnosed with severe tonsillar hypertrophy display discernible craniofacial features distinct from those with adenoid hypertrophy, prompting illuminating considerations regarding microbiota regulation in this non-inflammatory condition. The present study aimed to characterize the salivary microbial profile in children with tonsillar hypertrophy and explore the potential functionality therein. A total of 112 children, with a mean age of 7.79 ± 2.41 years, were enrolled and divided into the tonsillar hypertrophy (TH) group ( n = 46, 8.4 ± 2.5 years old), adenoid hypertrophy (AH) group ( n = 21, 7.6 ± 2.8 years old), adenotonsillar hypertrophy (ATH) group ( n = 23, 7.2 ± 2.1 years old), and control group ( n = 22, 8.6 ± 2.1 years old). Unstimulated saliva samples were collected, and microbial profiles were analyzed by 16S rRNA sequencing of V3-V4 regions. Diversity and composition of salivary microbiome and the correlation with parameters of overnight polysomnography and complete blood count were investigated. As a result, children with tonsillar hypertrophy had significantly higher α-diversity indices ( P ＜0.05). β-diversity based on Bray-Curtis distance revealed that the salivary microbiome of the tonsillar hypertrophy group had a slight separation from the other three groups ( P ＜0.05). The linear discriminant analysis effect size (LEfSe) analysis indicated that Gemella was most closely related to tonsillar hypertrophy, and higher abundance of Gemella , Parvimonas , Dialister , and Lactobacillus may reflect an active state of immune regulation. Meanwhile, children with different degrees of tonsillar hypertrophy shared similar salivary microbiome diversity. This study demonstrated that the salivary microbiome in pediatric tonsillar hypertrophy patients had different signatures, highlighting that the site of upper airway obstruction primarily influences the salivary microbiome rather than hypertrophy severity.IMPORTANCETonsillar hypertrophy is the most frequent cause of upper airway obstruction and one of the primary risk factors for pediatric obstructive sleep apnea (OSA). Studies have discovered that children with isolated tonsillar hypertrophy exhibit different craniofacial morphology features compared with those with isolated adenoid hypertrophy or adenotonsillar hypertrophy. Furthermore, characteristic salivary microbiota from children with OSA compared with healthy children has been identified in our previous research. However, few studies provided insight into the relationship between the different sites of upper airway obstruction resulting from the enlargement of pharyngeal lymphoid tissue at different sites and the alterations in the microbiome. Here, to investigate the differences in the salivary microbiome of children with tonsillar hypertrophy and/or adenoid hypertrophy, we conducted a cross-sectional study and depicted the unique microbiome profile of pediatric tonsillar hypertrophy, which was mainly characterized by a significantly higher abundance of genera belonging to phyla Firmicutes and certain bacteria involving in the immune response in tonsillar hypertrophy, offering novel perspectives for future related research., Competing Interests: The authors declare no conflict of interest.

The present study aimed to investigate the pharyngeal and nasal microbiota composition in children with adenotonsillar hypertrophy (AH) and assess longitudinal alterations in both microbiota after a probiotic oral spray treatment. A cohort of 57 AH patients were enrolled and randomly assigned to the probiotic and placebo groups for a 5-month treatment course. Pharyngeal and nasal swabs were collected before and after treatment and analyzed by 16S rRNA-based metataxonomics and axenic cultures for pathobiont identification. 16S rRNA sequences from pharyngeal and nasal swabs of 65 healthy children (HC) were used as microbiota reference profiles. We found that the pharyngeal and nasal microbiota of AH children were similar. When compared to HC, we observed an increase of the genera Rothia , Granulicatella , Streptococcus , Neisseria , and Haemophilus , as well as a reduction of Corynebacterium , Pseudomonas, Acinetobacter, and Moraxella in both microbiota of AH patients. After probiotic treatment, we confirmed the absence of adverse effects and a reduction of upper respiratory tract infections (URTI). Moreover, the composition of pharyngeal microbiota was positively influenced by the reduction of potential pathobionts, like Haemophilus spp., with an increase of beneficial microbial metabolic pathways. Finally, the probiotic reduced the abundance of the pathobionts Streptococcus mitis and Gemella haemolysans in relation to AH severity. In conclusion, our results highlight the alterations of the pharyngeal and nasal microbiota associated with AH. Moreover, probiotic administration conferred protection against URTI and reduced the presence of potential pathobionts in patients with AH., Importance: Adenotonsillar hypertrophy (AH) is considered the main cause of breathing disorders during sleep in children. AH patients, after significant morbidity and often multiple courses of antibiotics, often proceed to tonsillectomy and/or adenoidectomy. Given the potential risks associated with these procedures, there is a growing interest in the use of nonsurgical adjuvant therapies, such as probiotics, that could potentially reduce their need for surgical intervention. In this study, we investigated the pharyngeal and nasal microbiota in patients with AH compared with healthy children. Furthermore, we tested the effects of probiotic spray administration on both disease symptoms and microbiota profiles, to evaluate the possible use of this microbial therapy as an adjuvant for AH patients., Competing Interests: Authors I.P. and G.M. are employed by GenomeUp SRL, Viale Pasteur, 6, 00144, Rome, Italy. Authors V.D. and L.M. are employed by DMG, Pomezia, Italy. Author L.P. received research support from DMG. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

BACKGROUND The adenoids are lymphatic tissue located in the nasopharynx and play a role in upper-airway immunity. Inflammation of the adenoids is called adenoiditis, which can cause a variety of symptoms. This is a common condition and is due to acute viral or bacterial infection. Most patients experience mild symptoms of upper-respiratory tract infection with a self-limiting course. CASE REPORT A 5-year-old female patient was brought into the clinic by her parents with concerns regarding hearing and sleep. Clinical assessment was consistent with persistent otitis media with effusion and sleep-disordered breathing. She was scheduled for surgery, including nasendoscopy, adenoidectomy, and bilateral grommet insertion. During surgery, direct visualization of the postnasal space showed complete obstruction by hypertrophic, inflamed adenoids covered in a thick, white film. A biopsy was taken, which detected herpes virus cytopathic effect. A diagnostic workup excluded a neoplastic process and other bacterial or fungal infections. A trial of oral antiviral medication was successful and follow-up nasendoscopy showed resolution of adenoid hypertrophy. CONCLUSIONS Direct visualization of the postnasal space, with a transoral mirror or 120-degree endoscope, prior to adenoidectomy can aid diagnosis. Adenoiditis may be caused by a wide range of organisms, including herpes virus. Active mucopurulent discharge should raise concern for infection by bacteria, fungi, or virus. Previous research on viral infection of the adenoids have been in asymptomatic patients with presumed latent infection and undergoing elective adenoidectomy. To our knowledge, this is the first paper to report on successful treatment with antiviral medication alone.

Background and Objectives : the upper respiratory tract harbors the highest bacterial density in the whole respiratory system. Adenoids, which are located in the nasopharynx , are a major site of bacterial colonies in the upper airways. Our goal was to use culture-independent molecular techniques to identify the breadth of bacterial diversity in the adenoid vegetations of children suffering from chronic rhinosinusitis and obstructive sleep apnea. Materials and methods : in total, 21 adenoid samples were investigated using amplification and sequencing of the V3-V4 hypervariable region of the bacterial 16S rRNA gene. Results : among the most common bacterial species found were Veillonella atypica , Fusobactrium nucelatum , Shaalia odontolytica , and Moraxella catarrhalis . Veillonella atypica and Fusbacterium nucelatum dominated the microbiome in all 21 samples, attributing to more than 60% of all detected genetic material. Conclusions : since both Veillonella atypica and Fusobacterium nucleatum are, predominantly, oral cavity and dental microorganisms, our findings may suggest oral microbiome migration deeper into the oropharynx and nasopharynx where these bacteria colonize adenoid vegetations.

Chlamydophila pneumoniae is an intracellular pathogen responsible for respiratory tract infections. The isolation of the microorganism from clinical specimens is essential for a diagnosis. However, the identification of C. pneumoniae by cell cultures is very difficult besides strongly depending on the sample conditions. The study aimed to investigate, in adult patients with pharyngotonsillitis, the frequency of Chlamydophila pneumoniae detection by cell cultures and three conventional PCRs (a conventional PCR targeting the 16S rRNA gene and two nested PCRs, targeting the 16S rRNA gene and the ompA gene, respectively). The presence of chlamydial inclusion in cell cultures was observed in 11/94 samples (11.70%) by IFA. C. pneumoniae DNA was detected in 12/94 (12.76%) specimens by the 16S rRNA gene nested PCR, 4/94 (4.26%) by ompA gene nested PCR, and in 2/94 (2.13%) by 16S rRNA single-step PCR. Our data show poor agreement between the three applied DNA-amplification methods; in fact, only 16S rRNA gene nested PCR showed a statistically significant difference. Moreover, this result allowed us to achieve a definitive confirmation of the previous finding and to avoid the risk of an overestimation of the C. pneumoniae as a pathogen in pharyngotonsillitis.

Objective: Interleukin-17 (IL-17) C is a cytokine expressed by epithelial cells in response to bacterial stimulation. In contrast to other members of the IL-17 family of cytokines, IL-17C is upregulated early during infection, maintains integrity of the epithelial layer barrier, and mediates the innate immune response. We investigated the expression profile of IL-17C in pediatric adenoids. Methods: Pediatric adenoid tissues and lavage fluids were collected from a total of 38 subjects. The Limulus amebocyte lysate test and real-time PCR using Staphylococcus aureus primers were performed to evaluate bacterial contents in adenoids. Expression of IL-17RE in adenoids was analyzed using real-time polymerase chain reaction and western blot. The expression of IL-17C was evaluated by western blot and immunohistochemistry and compared between allergic rhinitis (AR) and control subjects. The levels of Hsp27, Hsp70, and IL-17C in adenoid lavage fluids were evaluated by enzyme-linked immunosorbent assay, and the correlation between these molecules was statistically analyzed. Results: The pediatric adenoids were found to be exposed to bacteria and had a normal flora comprising both gram-negative and -positive bacteria. IL-17RE, an IL-17C specific receptor, was highly expressed in the epithelium of adenoids. IL-17C was expressed in all evaluated adenoid tissue samples, irrespective of the allergic status of the patient. IL-17C secretion was detected in half of the adenoid lavage fluid samples and was associated with Hsp70 level. Conclusion: Our findings indicate the possible role of pediatric adenoids in innate immunity modulation via an innate immunity-associated cytokine., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Introduction: Otitis media with effusion (OME) is one of the most common pediatric diseases worldwide. Several studies have analyzed the diversity of the microbiomes found in the middle ear effusions (MEEs) of populations from developed countries. However, no microbiological studies of MEEs from Chinese children with OME have been reported. This study investigated the middle ear and adenoid microbiological profiles of children with OME, and compared the microbial flora of the adenoid between children with and without otitis media., Methods: MEEs and adenoid swabs were acquired from 15 children undergoing ventilation tube insertion and adenoidectomy. Adenoid swabs from 15 patients with no ear disease were used as controls. Samples were analyzed by 16S rRNA sequencing. Operational taxonomic units (OTUs) abundance information were normalized. Alpha diversity analyses were used to assess the richness and diversity of the microbial community for each sample. Beta diversity analyses were used to determine the inter-group variability between microbiome structure., Results: Based on the mean relative abundance, the MEEs were dominated by Haemophilus (14.75%), Staphylococcus (9.37%) and Halomonas (7.85%), and the bacterial compositions of the adenoids in the OME groups were dominated by Haemophilus (21.87%), Streptococcus (19.65%), and Neisseria (5.8%). The bacterial compositions in the adenoids of the controls were dominated by Haemophilus (15.96%), Streptococcus (13.33%), and Moraxella (12.28%). Alpha diversity analyses showed that there were no significant differences in microbiome richness or diversity between the middle ear effusions (TM) and adenoids (TA) of OME subjects. Adenoid samples from OME patients (TA) and control patients (CA) were also similar. Beta diversity analyses showed that the microbiomes of the adenoids in OME patients were also similar to that of controls. However, the microbiome structure of middle ear effusions was dissimilar to those of the adenoids in OME patients according to beta diversity analyses., Conclusions: Our results confirmed the microbial diversity of MEEs among Chinese children. However, the dissimilar microbiome composition between samples taken from the surface of the adenoids and from the middle ear effusions challenges the conventional theory that the adenoid serves as a microbial reservoir in children with otitis media with effusion., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Mycobacterium tuberculosis (Mtb) can enter the body through multiple routes, including via specialized transcytotic cells called microfold cells (M cell). However, the mechanistic basis for M cell entry remains undefined. Here, we show that M cell transcytosis depends on the Mtb Type VII secretion machine and its major virulence factor EsxA. We identify scavenger receptor B1 (SR-B1) as an EsxA receptor on airway M cells. SR-B1 is required for Mtb binding to and translocation across M cells in mouse and human tissue. Together, our data demonstrate a previously undescribed role for Mtb EsxA in mucosal invasion and identify SR-B1 as the airway M cell receptor for Mtb., Competing Interests: HK, VN, CR, SA, JG, LF, LH, PS, JK, YX, RM, MS No competing interests declared, (© 2020, Khan et al.)

Background : We previously described that adenoid tissue in children with chronic otitis media (COM) contained more mucosal biofilms than adenoid tissue removed for hypertrophy. Aims/objectives: The aim of the second part was to characterize nasopharyngeal microbiota and explore virulence of the most common middle ear pathogens. Material and methods : Bacteriological analysis was performed following a culture-based approach on the samples recovered from 30 patients of COM group (15 biofilm-positive and 15 biofilm-negative) and from 30 patients of a control group (15 biofilm-positive and 15 biofilm-negative). Virulence factors of Streptococcus pneumoniae, Streptococcus pyogenes , and Haemophilus influenzae were investigated. Results : The most frequent species were Firmicutes followed by Proteobacteria and Actinobacteria . The presence of biofilm was statistically associated with an increase of the number of bacterial species and Firmicutes phylum regardless of the condition (case/control). No virulence factors associated with invasive isolates were found for the most common middle ear pathogens. Conclusions and significance : This case-control study demonstrated that the presence of COM plus biofilm was associated with a given microbiota which contained more Firmicutes. Our study allows a better understanding of physiopathological mechanisms involved in chronic otitis media and paves the way for further investigations.

Streptococcus pneumonia, one of the major colonizers in nasopharyngeal adenoids, has been the predominant pathogen causing acute otitis media (AOM) in children. Recent evidence suggests an association between IL-17A-mediated immune response and the clearance of pneumococcal colonization in nasopharyngeal adenoids. Here, we evaluated the expressions of IL-17A and associated genes in hypertrophic adenoid tissues of children with sleep-disordered breathing (SDB) and otitis media with effusion (OME) and their association with pneumococcal carriage. Sixty-six pediatric patients with adenoid hypertrophy were enrolled. During adenoidectomy, nasopharyngeal swab and adenoid tissues were used to determine pneumococcal carriage and IL-17A expression. Our results revealed significantly higher levels of IL-17A and IL-17A:IL-10 mRNA in the SDB patients positive for nasopharyngeal pneumococcal carriage than those negative. However, these differences were not significant in the OME group. These results suggested, in OME patients, prolonged or chronic pneumococcal carriage may occur because of insufficient IL-17A-mediated mucosal clearance, and could further lead to AOM and OME development.

Objectives: The adenoid pad, which is located between the orifice of the Eustachian tube (ET) and posterior nasal cavity, can affect the development of otitis media with effusion (OME) because of its anatomical location. The aim of the present study was to evaluate adenoid microbial colonization through 16S ribosomal RNA (rRNA) pyrosequencing, an advanced molecular technique, and to document the relationship with OME., Materials and Methods: Adenoid samples were collected using sterile cotton from 32 children during ventilation tube insertion. Sixteen children with OME who underwent tonsillectomy and adenoidectomy due to obstructive symptoms were assigned to the OME group and sixteen children without OME were assigned to the control group. We performed a 16S rRNA-based culture-independent survey of bacterial communities using the MiSeq platform., Results: The diversity index, mean operational taxonomic units, and Shannon index were lower in the OME group than those in the control group. A taxonomic analysis showed differences in microbiota distribution between the OME and control groups at the phylum, genus, and species levels. The analysis, which was based on weighted UniFrac distances, revealed differences in microbial composition between the two groups., Conclusion: Bacterial community analysis using 16S rRNA pyrosequencing allows us to understand the relationship between the microbial communities of adenoids and the development of OME better.

Objective: The aim of this study was to evaluate the composition and the diversity of bacteriome in middle ear effusion (MEE) and adenoid specimens of pediatric patients having otitis media with effusion (OME)., Materials and Methods: Sample collection from children with OME followed by next generation sequencing. Seventeen adenoid and 43 middle ear effusion specimens from 25 children having OME were evaluated. Microbiome analysis was performed via Ion 16S rRNA metagenomics kit., Results: Twenty-two different bacterial species were identified from all of the samples analyzed. There were variations in the prevalence and relative abundance of the bacteriome observed between adenoid and MEE samples. MEE microbiome was significantly dominated by Alloicoccus otitis (44%), Turicella otitidis (6%), and Staphylococcus auricularis (3%). Whereas, Rothia mucilaginosa (39%), R. dentocariosa (11%), S. aureus (5%), Veillonella rogosae (2%), Granulicatella elegans (2%), Granulicatella adiacens (2%), Eikenella corrodens (1%), and Prevotella nanceiensis (1%) had significantly higher relative abundance in adenoid samples. Overall, there was no statistically significant difference in alpha diversity of MEE and adenoid samples, whereas adenoid samples constituted a cluster in the beta diversity graph., Conclusion: Bacteriome of MEE is mostly dominated by A. otitis yet accompanied by other bacteria with lower relative abundances suggests that OME is likely to be a polymicrobial process. Despite similarities, significant differences in relative abundances of several predominant species between bacteriome in the MEE and adenoid put the theory that OME in children is originated from the adenoids under question., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Biofilms are communities of bacteria embedded in a self-produced glycocalyx matrix. Adenoids have been shown to harbor bacterial biofilms. Aim/objectives: To compare the prevalence of biofilms in adenoid of children with chronic otitis media (COM) (group1) versus a control group without any COM (group 2) having adenoids removed because of hypertrophy., Material and Methods: One hundred and three children were prospectively enrolled in this case-control study, group 1 (n = 52) and group 2 (n = 51). The main outcome measurement was the prevalence of biofilm in adenoidectomy specimens analyzed using confocal laser scanning microscopy. Children in group 1 who had middle ear (ME) effusion and requiring the insertion of a tympanostomy tube underwent biopsy of the ME mucosa and effusion sampling., Results: Biofilms were found in adenoids' specimens of both groups and in the ME biopsy and effusion. The biofilm prevalence in adenoids was 63.5% (33/52) in group 1 and 47.1% (24/51) in group 2. Day nursery and previous antibiotics intake were significantly more frequent in group 1 than in group 2., Conclusions and Significance: This case-control study demonstrates that adenoid tissue in children with COM contains more mucosal biofilms than adenoid tissue removed for hypertrophy. Biofilm was seen in ME biopsies and effusion.

Objective: Adenoidectomy is a surgical procedure with potential adverse events. Effective nonsurgical therapy could reduce patient risk and harm. The aim of this study was to evaluate the role of bacteriotherapy to reduce the necessity of adenoid surgery., Patients and Methods: This experimental study was conducted as an open study in 44 children (30 males and 14 females, mean age 4.9 years) who were candidates for adenoidectomy and tympanocentesis as treatment for adenoidal hypertrophy and otitis media with effusion. Twenty-two children were treated with Streptococcus salivarius 24SMB and Streptococcus oralis 89a nasal spray, administered as 2 puffs per nostril twice a day for a week for 3 months (study group). The other half of the children was treated with hypertonic saline nasal lavage on the same schedule (control group). Tympanometry and adenoid size assessment were evaluated throughout the intervention period., Results: In the study group, 6/22 children required surgery, compared to 20/22 children in the study group (p<0.0001). The clinical change in the treated children was a significant reduction of adenoid size (p<0.0001) and improvement of middle ear effusion measured with tympanometry (p<0.0001)., Conclusions: Bacteriotherapy with Streptococcus salivarius 24SMB and Streptococcus oralis 89a nasal spray could significantly reduce the need for adenoid surgery.

Introduction: Adenotonsillar and middle ear diseases result in some of the most frequently performed operations in the pediatric population worldwide. The pathogen reservoir hypothesis (PRH) suggests that the adenoids act as a reservoir of bacteria which play a potential pathogenic role in otitis media. Evidence supporting this hypothesis is limited. This study sought to comprehensively determine and compare associations between the adenotonsillar and middle ear bacterial microbiota within individual patients via next-generation sequencing and microbial network analyses., Methods: Bacterial 16S rRNA gene-targeted amplicon sequencing was used to determine the bacterial composition of ten pediatric patients undergoing adenotonsillectomy and ventilation tube insertion for otitis media with effusion. At the time of surgery, swabs were taken from the adenoid surface, tonsil crypts and middle ear clefts (through the myringotomy incision)., Results: The most abundant sequences within the bacterial community at genus level across all anatomical sites were Fusobacterium, Haemophilus, Neisseria, and Porphyromonas. There was an observable difference in the relative abundance of bacterial communities, with a higher proportion of Haemophilus and Moraxella in the adenoid when compared with the middle ear. Furthermore, only one module (consisting of 4 bacterial OTUs) from one patient was identified through microbial network analyses to be significantly associated between middle ear and adenoid. In addition, microbial network analysis revealed that the adenoid and tonsil microbiota share greater similarity than do the adenoid and middle ear., Conclusion: The results of this study suggest that the adenoid microenvironment does not correlate to the middle ear microenvironment. A future study at the species level, and over time, is required to further investigate whether the differing relationship between the microbiota of the adenoid and middle ear rejects the pathogen reservoir hypothesis., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Tonsil and adenoid-tissue hypertrophy (AH) is the most common cause of pediatric sleep-disordered breathing (SDB), with AH possibly initiated by repeated exposure to infectious agents or allergens. Here, we evaluated IL-17A activity in adenoid tissue from children with SDB and its association with AH and pneumococcal carriage. Thirty-five children (aged 3-12 years) with SDB and receiving adenoidectomy and tonsillectomy were enrolled. During surgery, nasopharyngeal carriage was determined by bacterial culture and multiplex PCR via nasopharyngeal swab, and adenoid samples were collected. IL-17A and associated cytokine expression was evaluated by real-time PCR and western blotting. The mRNA analysis showed that IL-17A level, IL-17A:IL-10 ratio, and RAR-related orphan receptor-γt:forkhead box P3 ratio were significantly higher in adenoid tissues with AH, as were IL-17A level and IL-17A:IL-10 ratio in adenoid tissues with pneumococcal carriage. Additionally, pneumococcal carriage was more common in nasopharyngeal adenoids from patients without AH than those with AH. IL-17A was upregulated in adenoid tissues from patients with AH and with pneumococcal carriage. These results suggested that pneumococcal carriage initiates an IL-17A-mediated immune response in nasopharyngeal adenoids, which might be associated with AH in patients with SDB.

Introduction: Adenoids are nasopharyngeal lymphoid tissue with a relevant role in host defence against infection of upper respiratory tract. Nevertheless, adenoids are also a reservoir of microorganisms that can cause infections of upper respiratory tract and otitis particularly in children., Objective: Evaluate and compare the association between biofilm assembly on adenoids and the incidence of recurrent infections in a paediatric population submitted to adenoidectomy by either infectious or non-infectious indication., Methods: Scanning electron microscopy was used to assess biofilms on adenoid surface; biofilm assembly in vitro was monitored by crystal violet assay; antibiotic susceptibility was assessed following EUCAST guidelines; Hinfluenzae capsular typing was performed by PCR., Results: Biofilms were present in 27.4% of adenoid samples and no statistical difference was found between infectious and non-infectious groups. In vitro, the most clinically relevant bacteria, H.influenzae, S.aureus, S.pyogenes, S.pneumoniae and M.catarrhalis, were mostly moderate biofilm assemblers (71.7%). 55.3% of these bacteria were intermediate/resistant to at least one of the tested antibiotics. No association was found between the ability to assemble biofilms in vitro and the presence of biofilms on adenoids nor antibiotic resistance. All H.influenzae were characterized as non-typeable., Conclusion: The presence of biofilms on adenoid surface was independent from clinical sample background. Bacterial ability to assemble biofilms in vitro cannot be used to predict biofilm assembly in vivo. The lack of correlation between biofilm formation and infectious respiratory diseases found contributes to question the relevance of biofilms on the pathogenesis of infectious diseases.

Introduction: Culture-independent methods, based on bacterial 16 S rRNA gene sequencing, have been used previously to investigate the adenotonsillar microbiota. However, these studies have focused on a single sampling site (usually a surface swab). We aimed to investigate potential differences in adenotonsillar microbiota according to sampling location, both on and within the adenoids and palatine tonsils., Methods: Pediatric patients (n = 28, mean age five years) undergoing adenotonsillectomy were recruited for this study. At the time of surgery, a mucosal adenoid surface swab and an adenoid tissue biopsy was collected. Immediately following surgery, the crypts of the right and left tonsils were swabbed, and a surface and core tissue sample from the right tonsil were also collected. Bacterial 16 S rRNA gene-targeted amplicon sequencing was used to determine the bacterial composition of the collected samples., Results: There was no significant difference in diversity or composition of the adenoid microbiota based on sampling site. However, the Shannon-Wiener and Inverse-Simpson diversity indices differed significantly (p < 0.05) between the microbial communities of the three different tonsil sampling sites. There was a higher average relative abundance of members from the genera Streptococcus, Actinobacillus, and Neisseria in the tonsil crypts when compared with surface and core tonsil tissue samples., Conclusion: Our results indicate that there is variation in bacterial diversity and composition based on sampling sites in the tonsils but not the adenoids. The difference in microbiota between the surface and the tissue may have implications for our understanding of the pathogenesis of recurrent tonsillitis and have treatment implications., (Copyright © 2018. Published by Elsevier B.V.)

Exposure to tobacco smoke is associated with a higher risk of respiratory tract diseases. The aim of this study was to determine the influence of passive smoking on selected characteristics of children with adenoid hypertrophy. Sixty-one children with adenoid hypertrophy were enrolled in the prospective study. Differences in bacterial colonisation of middle nasal meatus and nasopharynx and changes in selected laboratory immune and inflammatory markers according to the tobacco smoke exposure were analysed. Exposure to tobacco smoke was associated with significantly higher colonisation of pathogenic bacteria and polymicrobial growth of pathogenic bacteria (≥ 2 bacteria) in middle nasal meatus compared to non-exposed children (P = 0.045, P = 0.032, respectively). Identification of pathogenic bacteria in the middle nasal meatus did not correlate with isolation of pathogenic bacteria in the nasopharynx in either group of children. Parameters of humoral immunity in serum, IgA and IgG, were detected at higher concentrations in children exposed to tobacco smoke (P = 0.047, P = 0.031, respectively). Differences in selected parameters of cellular immunity in peripheral blood according to passive smoking were not observed. Tobacco smoke exposure is related to increased colonisation by pathogenic bacteria in middle nasal meatus and elevation of IgA and IgG in peripheral blood, but does not seem to influence markers of cellular immunity parameters in children with adenoid hypertrophy. Avoidance of passive smoking could be recommended as a universal preventive strategy against microbial colonisation of the upper airways and development of various inflammatory diseases in children, e.g. adenoid hypertrophy., (Copyright © 2018 Società Italiana di Otorinolaringologia e Chirurgia Cervico-Facciale, Rome, Italy.)

Pathogenic bacteria associated with the adenoids and tonsils cause much morbidity in the paediatric population. Hyperplasia of the adenoids is associated with otitis media with effusion and hyperplasia of the palatine tonsils is associated with both recurrent tonsillitis and obstructive sleep apnoea. Most current knowledge of the microbiology of the upper airways has been derived from culture-based studies, which usually reflect only a small fraction of the bacteria present on the mucosal surface. Culture-independent molecular surveys based on 16S ribosomal RNA sequencing are now being employed to determine the microbiota on the surface and within the tissue of adenoids and palatine tonsils. This review describes the new techniques applied in determining the microbiome and summarises the results of studies employing these techniques., Competing Interests: Competing interests: None declared., (© Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2018. All rights reserved. No commercial use is permitted unless otherwise expressly granted.)

This paper attempts to identify the sources and evaluate the prevalence of halitosis in children with adenoid hypertrophy. The study included pediatric patients admitted for adenoidectomy due to obstructive symptoms. Patients with possibly other causes of halitosis, were excluded from the study. Halitosis was detected in 30 out of 136 children (22.1%). The effect of adenoid hypertrophy on halitosis was confirmed by the significant reduction in the organoleptic score rated according to the Rosenberg scale (on average by 2°) and the levels of volatile sulfur compounds (on average 84 ppb) post-adenoidectomy, as compared to pre-surgery results. In total, we reported a statistically significant decrease in the abovementioned values in 90% of patients with halitosis. It was found that in patients with halitosis, as compared to patients with no oral malodor, anaerobic bacteria and Staphylococcus aureus were more frequently observed in the adenoid tissue. Streptococcus oralis strains were more commonly reported in the material taken during adenoidectomy in children without oral malodor.

Objective: To compare the microbial flora of adenoids from patients with recurrent acute otitis media (AOM) and patients with obstructive sleep apnea (OSA)., Materials and Methods: This study was prospective and controlled. Adenoids were obtained from children undergoing adenoidectomy for recurrent AOM (n=7) or OSA (n=13). Specimens were processed for total deoxyribonucleic acid (DNA) isolation. 16s DNA 454-pyrosequencing was performed on AOM (n=5) and OSA (n=5) specimens. All specimens were analyzed by real-time polymerase chain reaction for the quantification of the oral commensal bacteria, Streptococcus salivarius., Results: All adenoid specimens had evidence of microbes. Haemophilus influenzae, Moraxella catarrhalis, Streptococcus pneumoniae, Pseudomonas aeruginosa, and Staphylococcus aureus were among the dominant species in all samples. S. salivarius, Prevotella sp. and Terrahaemophilus aromaticivorans were more common on adenoids from OSA patients (p<0.05). Bradyrhizobium sp. was more common on adenoids from patients with recurrent AOM (p<0.05). The microbial profiles associated with recurrent AOM were different from, but overlapped with OSA. S. salivarius quantified by real-time PCR was not different between the two groups., Conclusion: Microbes are present on all adenoid specimens, though the microbial profile differs between recurrent AOM and OSA. The clinical significance of these differences remains to be determined.

Adenoids play a key role in both respiratory and ear infection in children. It has also been shown that adenoidectomy improves these symptoms in this population. The main goal of the present study was to evaluate adenoid bacterial colonization and document a possible relation with infectious respiratory disease. A prospective observational study was designed to evaluate the proposed hypothesis in a paediatric population submitted to adenoidectomy by either infectious or non-infectious indications and compare these two cohorts. A total of 62 patients with ages ranging from 1 to 12 years old were enrolled in the study. Adenoid surface, adenoid core and middle meatus microbiota were compared. A close association between adenoid colonization and nasal infection was found, supporting that adenoids may function as bacterial reservoir for upper airway infection. The obtained results also contribute to explain the success of adenoidectomy in patients with infectious indications.

Objectives: To evaluate the presence of viruses and bacteria in middle ear and adenoids of patients with and without otitis media with effusion (OME)., Methods: Adenoid samples and middle ear washes (MEW) were obtained from children with OME associated with adenoid hypertrophy undergoing adenoidectomy and tympanostomy, and compared to those obtained from patients undergoing cochlear implant surgery, as a control group. Specific DNA or RNA of 9 respiratory viruses (rhinovirus, influenza virus, picornavirus, syncytial respiratory virus, metapneumovirus, coronavirus, enterovirus, adenovirus and bocavirus) and 5 bacteria (S. pneumoniae, H. influenzae, M. catarrhalis, P. aeruginosa and S. aureus) were extracted and quantified by real-time PCR., Results: 37 OME and 14 cochlear implant children were included in the study. At the adenoid, virus and bacteria were similarly detected in both OME and control patients. At the middle ear washes, however, a higher prevalence of bacteria was observed in patients with OME (p = 0.01). S. pneumoniae (p = 0.01) and M. catarrhalis (p = 0.022) were the bacteria responsible for this difference. Although total virus detection was not statistically different from controls at the middle ear washes (p = 0.065), adenovirus was detected in higher proportions in adenoid samples of OME patients than controls (p = 0.019)., Conclusions: Despite both OME and control patients presented similar rates of viruses and bacteria at the adenoid, children with OME presented higher prevalence of S. pneumonia, M. catarrhalis in middle ear and adenovirus in adenoids when compared to controls. These findings could suggest that these pathogens could contribute to the fluid persistence in the middle ear.

Importance: The adenoid pad has long been considered a reservoir for bacteria in the pathogenesis of otitis media with effusion (OME). However, bacteria more reminiscent of external auditory canal (EAC) commensals are often demonstrated within middle ear aspirates., Objective: To compare the microbiota of the EAC, the middle ear with OME, and the adenoid pad to further clarify the true source of middle ear bacteria., Design, Setting, and Participants: Middle ear fluid (MEF) aspirates and EAC lavages were collected from 18 children with OME undergoing ventilation tube insertion from June 1, 2014, to August 31, 2015, at Women and Children's Hospital, Adelaide, Australia. Adenoid pad and MEF samples were included from a previous study. Samples were analyzed using sequencing of the 16S ribosomal RNA gene. Previously collected microbiota data from the adenoid pad were collated for analysis., Main Outcome Measures: Mean relative abundance of top bacterial genera for the MEF, EAC, and adenoid pad samples., Results: Eighteen pediatric patients with chronic OME (6 female; 12 male; mean [SD] age, 48 [36] months) were recruited prospectively, with 34 paired MEF and EAC samples. The MEF microbiota (mean relative abundance [SD]) consisted of Alloiococcus otitidis (37.5% [40.0%]), Haemophilus (14.4% [29.1%]), Moraxella (10.0% [26.4%]), Staphylococcus (8.2% [21.9%]), and Streptococcus (3.8% [13.1%]). The mean relative abundance (SD) microbiota of the EAC demonstrated a sparsity of classic otopathogens, including Haemophilus (0.3% [0.8%]), Moraxella (0.3% [0.7%]), and Streptococcus (0.2% [0.6%]), but had a high abundance of Alloiococcus (58.0% [44.1%]), Staphylococcus (20.8% [34.0%]), and Pseudomonas (3.2% [17.1%]). In contrast, based on previously collected data, the microbiota of the adenoid pad showed a high abundance of the classic otopathogens with a sparsity of EAC genera for Alloiococcus (0.1% vs 28.9%, respectively; P < .001), Haemophilus (25.2% vs 18.2%, respectively; P = .002), Staphylococcus (0.2% vs 10.8%, respectively; P = .02), Streptococcus (12.7% vs 4.2%, respectively; P < .001), and Pseudomonas (0 vs 2.1% respectively; P < .001). The microbiota of the MEF collected during 2 consecutive years were similar (Alloiococcus, 22.7% vs 37.5%; Haemophilus, 22.5% vs 14.0%; Staphylococcus, 10.9% vs 10.7%; Moraxella, 5.0% vs 9.7%; Corynebacteria, 6.2% vs 3.1%; Streptococcus, 4.8% vs 3.7%; and Pseudomonas, 1.1% vs 3.0%; P ≥ .05)., Conclusions and Relevance: The EAC and the nasopharynx could serve as reservoirs for microbiota of the middle ear. Furthermore, the microbiota of the middle ear with effusion appear to be relatively stable over time and between populations with OME.

Objectives/hypothesis: The adenoid pad has been considered a reservoir for bacteria in the pathogenesis of otitis media with effusion. This study aimed to characterize the middle ear microbiota in children with otitis media with effusion and establish whether a correlation exists between the middle ear and adenoid microbiota., Study Design: Prospective, controlled study., Methods: Middle ear aspirates adenoid pad swabs were collected from 23 children undergoing ventilation tube insertion. Adenoid swabs from patients without ear disease were controls. Samples were analyzed using 16S rRNA sequencing on the Illumina MiSeq platform., Results: Thirty-five middle ear samples were collected. The middle ear effusion microbiota was dominated by Alloiococcus otitidis (23% mean relative abundance), Haemophilus (22%), Moraxella (5%), and Streptococcus (5%). Alloiococcus shared an inverse correlation with Haemophilus (P = .049) and was found in greater relative abundance in unilateral effusion (P = .004). The microbiota of bilateral effusions from the same patient were similar (P < .001). However, the otitis media with effusion microbiota were found to be dissimilar to that of the adenoid (P = .01), whereas the adenoid microbiota of otitis media with effusion and control patients were similar (P > .05) (permutational multivariate analysis of the variance)., Conclusions: Dissimilarities between the local microbiota of the adenoid and the middle ear question the theory that the adenoid pad is a significant reservoir to the middle ear in children with otitis media with effusion. A otitidis had the greatest cumulative relative abundance, particularly in unilateral effusions, and shares an inverse correlation with the relative abundance of Haemophilus., Level of Evidence: NA Laryngoscope, 126:2844-2851, 2016., (© 2016 The American Laryngological, Rhinological and Otological Society, Inc.)

Biofilms have been implicated in the development of several chronic upper respiratory tract infections. Role of bacterial biofilms has been well studied in the pathogenesis of chronic rhinosinusitis. However, its impact on development of middle ear effusion is still a matter of debate. To study the extent of surface adenoid biofilm and evaluate its role in the pathogenesis of chronic otitis media with effusion in children. The study was carried out on 40 children in Alexandria Main University Hospital between 1 and 16 years of age without sex predilection, who were divided into two groups. The first group (20 children) had otitis media with effusion associated with adenoid hypertrophy, whereas the second group (20 children) had adenoid hypertrophy without middle ear effusion. Adenoidectomy with ventilation tube insertion was done for group 1 cases, whereas, only Adenoidectomy was done for group 2 cases. The samples were processed for the detection of biofilms by scanning electron microscopy. The biofilm formation was graded according to extension. Biofilm formation was detected on all samples for group 1. Adenoids removed from patients with otitis media with effusion had higher-grade biofilm formation than the other group (P 0.0001). No correlation was found between adenoid size and biofilm formation. In pediatric population, adenoid surface biofilm formation may be involved in the pathogenesis otitis media with effusion.

Otitis media with effusion (OME) is a highly prevalent disease in children, but the exact pathogenesis and role of bacteria are still not well understood. This study aimed to investigate the presence of otopathogenic bacteria in the middle ear effusion (MEE) and adenoid of children with chronic OME (COME), and to investigate in vivo whether these bacteria, especially Haemophilus influenzae, are organized as a biofilm in the middle ear fluid. MEE and adenoid samples were collected from 21 patients with COME. Extensive bacterial culturing and genotyping was performed on all middle ear and adenoid samples. Fluorescence in situ hybridization (FISH) and confocal laser scanning microscopy (CLSM) was used to visualize possible biofilm structures for a selection of middle ear effusion samples. 34 MEE samples were collected from 21 patients of which 64.7 % were culture positive for bacteria and 47.0 % were culture positive for Haemophilus influenzae, Moraxella catarrhalis, Staphylococcus aureus and/or Streptococcus pneumoniae. All 21 adenoid samples were culture positive for one or more of these four otopathogens. H. influenzae (35.3 %) and S. pneumoniae (76.2 %) were the most frequently cultured bacteria in the MEE and adenoid samples, respectively. The same bacterial species was found in MEE and adenoid for 84.6 % of the patients and in 81.2 % of the cases where the same species was found in more than one site it involved the same bacterial genotype. FISH and CLSM demonstrated the presence of H. influenzae specific biofilm structures in five of the eight culture positive MEEs that were tested, but in none of the two culture negative MEEs. The findings in this study indicate that the adenoid acts as a reservoir for bacteria in MEE and confirms that biofilms, in at least half of the cases consisting of H. influenzae, are indeed present in the MEE of children with COME. Biofilms may thus play a crucial role in the pathogenesis of COME, which is important in the understanding of this disease and the development of potential future treatment options.

Objective: A tropism to epithelial cells and lymphocytes, an inhibition of apoptosis in host cells, an ability to occurrence in persistent form resistant to antibiotic treatment are the features of Chlamydia pneumoniae, which can have connection with chronic inflammation of an adenoid tissue and adenoid hypertrophy. This study aimed to (1) detect the C. pneumoniae in an adenoid in children undergoing adenoidectomy, (2) estimate a connection between C. pneumoniae occurrence and the size of adenoid, (3) demonstration in which of adenoid cells C. pneumoniae occurs most often., Material and Methods: The examined group consisted of 200 children aged from 2 to 16 years (mean age 6,4) undergoing adenoidectomy. In all children during qualification for adenoidectomy a fiberoscopic examination of the nasopharynx was performed. A part of removed adenoid tissue was analysed by real-time PCR for C. pneumoniae. Adenoids from children with positive PCR examination and from 10 children with negative PCR examination were examined using immunohistochemistry (IHC)., Results: C. pneumoniae in the adenoid was present in 5,5% children. Positive results were obtained most frequently (24,14%, 7/29) in the eldest group (10-16 years). A statistical analysis demonstrated the correlation between C. pneumoniae occurrence in an adenoid tissue and the size of adenoid. In immunohistochemistry C. pneumoniae was found the most frequently in lymphocytes and in epithelial cells., Conclusions: A presence of C. pneumoniae in lymphocytes and epithelial cells of the adenoid first of all in older children with adenoid hypertrophy confirms the participation of this bacteria in adenoid pathology.

Background: We aimed to investigate the potentially pathogenic bacteria of upper respiratory tract infections (URTIs) and their susceptibilities to different antibiotics., Methods: Two-hundred adenoid and tonsil specimens from 100 patients who had undergone adenotonsillectomy were obtained and analyzed bacteriologically. Identification of the pathogens was made by conventional or commercial identification systems and antibiotic susceptibility tests were carried out by disk diffusion method., Results: A total of 274 pathogens were recovered from 81% specimens of 73% of the patients. Haemophilus influenzae (31.8%) was the most prevalent pathogen, followed by Staphylococcus aureus (17.2%), Group A beta hemolytic Streptococci, GABHS (12.0%), Moraxella catarrhalis (7.7%), Streptococcus pneumoniae (7.3%), and nine other bacterial species (24.0%). Penicillins (penicillin, ampicillin) had 100% activity against GABHS followed by 96.5% in H. influenzae, 45% in S. pneumoniae, and 0% in S. aureus strains. The efficacy of beta-lactamase inhibitor antibiotics (ampicillin/sulbactam, amoxycillin/clavulanic acid) were similar to those of penicillins but had superior activity (89.4%) against S. aureus strains. Cefotaxime had high activity (100%) against GABHS and H. influenzae followed by S. aureus (89.4%). Cotrimoxazole was also active in S. aureus (97.8%) and H. influenzae (83.9%) but revealed intermediate activity (45%) in S. penumoniae and was not efficient (0%) in GABHS. Macrolids (erythromycin, clindamycin) were very efficient (100%) in GABHS followed by S. aureus (95.7%) and had intermediate activity (50%) in S. pneumoniae. Levofloxacin, telithromycin, and vancomycin had 100% activity against S. pneumoniae strains., Conclusions: Our finding have shown that H. influenzae was the most prevalent pathogen followed by S. aureus, GABHS, M. catarrhalis, and S. pneumoniae and that there was no unique antibiotic to combat all prevalent pathogens, but penicillins could be the choice in GABHS and H. influenzae; beta-lactamase inhibitors and cefotaxime for GABHS, H. influenzae, and S. aureus; macrolids in GABHS and S. aureus; cotrimoxazole in H. influenzae and S. aureus; and levofloxacin and telithromycin in the treatment of S. penumoniae related URTIs.

Streptococcus pneumoniaeis one of the key pathogens responsible for otitis media (OM), the most common infection in children and the largest cause of childhood antibiotic prescription. Novel therapeutic strategies that reduce the overall antibiotic consumption due to OM are required because, although widespread pneumococcal conjugate immunization has controlled invasive pneumococcal disease, overall OM incidence has not decreased. Biofilm formation represents an important phenotype contributing to the antibiotic tolerance and persistence ofS. pneumoniaein chronic or recurrent OM. We investigated the treatment of pneumococcal biofilms with nitric oxide (NO), an endogenous signaling molecule and therapeutic agent that has been demonstrated to trigger biofilm dispersal in other bacterial species. We hypothesized that addition of low concentrations of NO to pneumococcal biofilms would improve antibiotic efficacy and that higher concentrations exert direct antibacterial effects. Unlike in many other bacterial species, low concentrations of NO did not result inS. pneumoniaebiofilm dispersal. Instead, treatment of bothin vitrobiofilms andex vivoadenoid tissue samples (a reservoir forS. pneumoniaebiofilms) with low concentrations of NO enhanced pneumococcal killing when combined with amoxicillin-clavulanic acid, an antibiotic commonly used to treat chronic OM. Quantitative proteomic analysis using iTRAQ (isobaric tag for relative and absolute quantitation) identified 13 proteins that were differentially expressed following low-concentration NO treatment, 85% of which function in metabolism or translation. Treatment with low-concentration NO, therefore, appears to modulate pneumococcal metabolism and may represent a novel therapeutic approach to reduce antibiotic tolerance in pneumococcal biofilms., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Objective: To determine Helicobacter pylori presence on antral lavages, adenoids and salival inmunoglobuline A on paediatric patients with chronic rhinosinusitis without nasal polyps (CRSsNP) and adenoid hypertrophy., Methods: Adenoid tissue, liquid obtained from antral lavages and saliva from 28 children diagnosed with CRSsNP, from the paediatric otorhinolaryngology practice of "Dr. Domingo Luciani" Hospital was taken and processed by means of polymerase chain reaction (PCR) using cagA, vacA and babA primers, also anatomopathological examination using Giemsa stain of the adenoids, determination of salivary specific secretory inmunoglobuline A (sIgA), socio-economic condition using the Graffar scale and associated gastrointestinal symptoms were assessed., Results: No evidence of Helicobacter pylori neither in antral lavages liquid nor adenoid tissue was found using PCR and Giemsa stain. sIgA was present in 28.6% of the subjects. The most frequently found symptoms were, diarrhea in 17.9%, distension and abdominal pain in 10.7%, 64.3% of the patients were in working (28.6%) and low middle (35.7%) classes., Conclusions: Helicobacter pylori is not present neither in maxillary sinuses nor adenoid tissue of the evaluated patients, sIgA it is a non-invasive method for assessment of immunologic challenge with the bacteria, not the presence of acute or chronic infection., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

The objective of the present retrospective study based on the analysis of 791 medical cards was to investigate microflora localized at the pharyngeal tonsil surface in the children living in the city of Chelyabinsk and presenting with exacerbation of chronic adenoiditis. 66% of the patients with this condition were 4-6 year old children. The most commonly documented were the associations composed of resident species S. alpha-haemoliticus and S. epidermidis together with pathobiontic S. aureus. S. beta-haemoliticus and S. pneumoniae occurred most frequently whereas H. influenza and some representatives of the genus Enterobacteriaceae were less numerous. Fungi of the genus Candida were identified in 25.4% of the samples at a titer above 10^4. In 23.7% of the cases, these fungi were found in the associations with S. aureus which resulted in mutual potentiation of the pathogenicity factor.

Adenoids as a first line of host defense against respiratory microbes play an important role in majority of upper airway infectious and noninfectious illnesses. Bacterial pathogen can colonize on the adenoid tissue and probably act as a reservoir for them. To determine phenotypic and genotypic characterization of biofilm forming capacity of Staphylococcus aureus isolates from children with adenoid hypertrophy and prevalence of Panton-Valentine leukocidin (PVL) gene we collected 17 consecutive, clinically significant S. aureus isolates from children with adenoid hypertrophy undergoing adenoidectomy with one or more of the upper airway obstruction symptoms, nasal obstruction, mouth breathing, snoring, or sleep apnea. Biofilm formation was evaluated by colorimetric microtiter plate's assay. Gene encoding PVL and adhesion- or biofilm formation-encoding genes were targeted by polymerase chain reaction (PCR) assay. According to the results, all strains produced biofilm. Seven (41.2%) isolates produced strong biofilm whereas 7 (41.2%) isolates produced week and 3 (17.6%) isolates produced medium biofilm. Regarding the adhesion- or biofilm formation-encoding genes, 16 (94.1%) isolates were positive for the gene eno, 13(76.4%) for icaA, 13 (76.4%) for icaD, 10 (58.8%) for fib, 10 (58.8%) for fnbB, 4(23.5%) for can, and 1(5.8%) for fnbA. The high prevalence of genes encoding biofilms and adhesins and phenotypic ability to form a biofilm by S. aureus strains emphasizes the pathogenic character of strains isolated from children with adenoid hypertrophy., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Introduction: Indigenous Australian children have a high prevalence of otitis media with effusion (OME) and associated conductive hearing loss. Only three microbiological studies of middle ear fluid (MEF) from Indigenous Australian children with OME have been reported. All of these were reliant on culture or species-specific PCR assays. The aim of this study was to characterise the middle ear fluid (MEF), adenoid and nasopharyngeal (NP) microbiomes of Indigenous Australian children, using culture-independent 16S rRNA gene sequencing., Methods: MEF, NP swabs and adenoid specimens were collected from 11 children in the Alice Springs region of Central Australia. Bacterial communities in these specimens were characterised using 16S rRNA gene sequencing., Results: The microbiota in MEF samples were dominated (>50% relative abundance) by operational taxonomic units (OTUs) consistent with Alloiococcus otitidis (6/11), Haemophilus influenzae (3/11) or Streptococcus sp. (specifically, Mitis group streptococci which includes Streptococcus pneumoniae) (1/11). Anatomical site selectivity was indicated by the presence of a single conserved Haemophilus OTU in 7/11 MEF samples. In comparison, there were ten distinct Haemophilus OTUs observed across the NP and adenoid samples. Despite significant differences between the MEF and NP/adenoid microbiomes, Streptococcus sp., H. influenzae and Moraxella catarrhalis OTUs were common to all sample types. Co-occurrence of classical otopathogens in paired MEF and NP/Adenoid samples is consistent with earlier culture-based studies., Conclusion: These data highlight the need to further assess H. influenzae traits important in otitis media and to understand the role of canal flora, especially A. otitidis, in populations with a high prevalence of tympanic membrane perforation., (Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.)

Conclusion: Lactobacillus rhamnosus GG (L.GG) was present in all adenoids of children receiving the L. GG product. However, since L.GG was also found from the placebo group, one cannot confirm its effect on the occurrence of rhinovirus (RV) or enterovirus (EV)., Objectives: The present study was conducted to determine whether a 3-week oral consumption of L.GG would lead to presence of the probiotic in adenoid tissue. Furthermore, nasopharyngeal RV and EV findings and symptom data were investigated., Method: The tissue samples were collected from 40 children aged 1-5 years about to undergo adenotomy due to recurrent acute/secretory otitis media, chronic rhinitis, or recurrent sinusitis after a 3-week daily consumption of L.GG (n = 20) or placebo (n = 20). Strain-specific real-time PCR was used to detect RV, EV, and L.GG in adenoid tissue., Results: L.GG was recovered in the adenoid sample in 100% of children in the L.GG group and in 76% in the placebo group (p = 0.07). Both RV and EV were found in 31% of children in the L.GG group and in 18% of children in the placebo group (p = 0.67). The majority of the positive samples were positive for both RV and EV. Study diaries showed no differences in symptoms between the groups.

Objective: To evaluate the role played by adenoids as a reservoir for infection in children assigned for adenoidectomy., Methodology: The study included 35 children with adenoid hypertrophy. All patients underwent clinical examination and adenoidectomy, adenotonsillectomy, or myringotomy with insertion of aeration tube according to indications. Surgical specimens were processed for conventional bacterial culture examination and to assay for biofilm formation. The obtained adherence values using spectrophotometer at 595 nm (OD595) was used to classify isolates according to its biofilm forming capacity., Results: We did adenotonsillectomy and myringotomy with insertion of aeration tube in 5 patients having adenotonsillitis with otitis media with effusion. We did adenotonsillectomy in 12 patients having adenotonsillitis and adenoidectomy in 18 patients having adenoid hypertrophy. Thirty-one surgical specimens showed bacterial growth on conventional media, while 4 specimens failed to give growth. The predominant organism was H influenzae then Staph aureus and Strept pneumoniae. Thirty-two specimens showed biofilm forming capacity (BFC) of variable extent, while others showed no BFC., Conclusion: Adenoids act as a bacterial reservoir secondary to bacterial biofilm formation so could induce chronicity and initiate development of complications. Determination of BFC using the proposed protocol is feasible, inexpensive, and available and spares the need for sophisticated instruments or approaches., (© The Author(s) 2015.)

Introduction: There is an ongoing debate about the existence and effects of Helicobacter pylori (Hp) in adenotonsillar tissue., Objective: A clinical study was conducted to assess the existence of Hp in the adenoid and/or adenotonsillar tissues, which were surgically excised due to chronic adenotonsillitis., Methods: Phosphoglucosamine mutase gene for the detection of Hp and cytotoxin-associated gene as virulence gene were examined in 84 adenotonsillar tissues obtained from 64 patients and patients' serum by using polymerase chain reaction., Results: Hp IgG was detected in 57 (89%) patients' serum. A total of seven tissue samples from 64 patients (10.9%) were found positive for Hp DNA, of which five were adenoids and two were tonsil tissues. All polymerase chain reaction positive samples were also positive for the cytotoxin-associated gene, which is a virulence determinant for the organism., Conclusion: This study suggests that children are exposed to Hp at an early age of their life in this province. Hp may have a role in the pathogenesis of chronic adenotonsillitis, especially in endemic areas., (Copyright © 2015 Associação Brasileira de Otorrinolaringologia e Cirurgia Cérvico-Facial. Published by Elsevier Editora Ltda. All rights reserved.)

Haemophili are pathogenic or opportunistic bacteria often colonizing the upper respiratory tract mucosa. The prevalence of Haemophilus influenzae (with serotypes distribution), and H. parainfluenzae in the nasopharynx and/or the adenoid core in children with recurrent pharyngotonsillitis undergoing adenoidectomy was assessed. Haemophili isolates were investigated for their ability to biofilm production.Nasopharyngeal swabs and the adenoid core were collected from 164 children who underwent adenoidectomy (2-5 years old). Bacteria were identified by the standard methods. Serotyping of H. influenzae was performed using polyclonal and monoclonal antisera. Biofilm formation was detected spectrophotometrically using 96-well microplates and 0.1% crystal violet.Ninety seven percent (159/164) children who underwent adenoidectomy were colonized by Haemophilus spp. The adenoid core was colonized in 99.4% (158/159) children, whereas the nasopharynx in 47.2% (75/159) children (P < 0.0001). In 32% (51/159) children only encapsulated (typeable) isolates of H. influenzae were identified, in 22.6% (36/159) children only (nonencapsulated) H. influenzae NTHi (nonencapsulated) isolates were present, whereas 7.5% (12/159) children were colonized by both types. 14.5% (23/159) children were colonized by untypeable (rough) H. influenzae. In 22% (35/159) children H. influenzae serotype d was isolated. Totally, 192 isolates of H. influenzae, 96 isolates of H. parainfluenzae and 14 isolates of other Haemophilus spp. were selected. In 20.1% (32/159) children 2 or 3 phenotypically different isolates of the same species (H. influenzae or H. parainfluenzae) or serotypes (H. influenzae) were identified in 1 child. 67.2% (129/192) isolates of H. influenzae, 56.3% (54/96) isolates of H. parainfluenzae and 85.7% (12/14) isolates of other Haemophilus spp. were positive for biofilm production. Statistically significant differences (P = 0.0029) among H. parainfluenzae biofilm producers and nonproducers in the adenoid core and the nasopharynx were detected.H. influenzae and H. parainfluenzae carriage rate was comparatively higher in the adenoid core than that in the nasopharynx in children undergoing adenoidectomy, suggesting that their involvement in chronic adenoiditis. The growth in the biofilm seems to be an important feature of haemophili colonizing the upper respiratory tract responsible for their persistence.

Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Staphylococcus aureus colonization of the adenoids and nasopharynx in 103 preschool children who underwent adenoidectomy for recurrent upper respiratory tract infections was examined. Bacterial interactions and risk factors for bacterial colonization of the nasopharynx and adenoids, separately, were analysed statistically. The prevalence of simultaneous isolation from both anatomical sites was 45·6% for S. pneumoniae, 29·1% for H. influenzae, 15·5% for M. catarrhalis and 18·4% for S. aureus. Three pathogens were significantly more frequent together from adenoid samples; nasopharyngeal swabs more often yielded a single organism, but without statistical significance. M. catarrhalis and S. aureus significantly more frequently co-existed with S. pneumoniae and H. influenzae than with each other and a positive association of S. pneumoniae and H. influenzae in adenoid samples was evident. Several differences between risk factors for nasopharyngeal and adenoid colonization by the individual pathogens were observed. We conclude that the adenoids and nasopharynx appear to differ substantially in colonization by pathogenic microbes but occurrence of H. influenzae and S. pneumoniae in the nasopharynx could be predictive of upper respiratory tract infections.

Background: Pneumococcal carriage in the nasopharynx is a primary means of transmission and a necessary prerequisite for pneumococcal disease., Objectives: We analyzed the relationship between expressions of Foxp3+ regulatory T (Treg) cells and Th17 cells, and pneumococcal carriage in the adenoids of children who were either positive or negative for pneumococci., Methods: We collected adenoidal tissue and nasopharyngeal swab samples from children undergoing an adenoidectomy. Adenoidal mononuclear cells were isolated, cultured and then stimulated with culture concentrated supernatant (CCS) obtained from a D39 bacterial strain., Results: Foxp3+ Treg cells were upregulated and Th17 cells were downregulated in populations of adenoidal mononuclear cells obtained from the pneumococcus-positive group. Following CCS stimulation, the increment in Foxp3+ Treg cells in the pneumococcus-positive group was significantly greater than that in the pneumococcus-negative group, while the increment in Th17 cells was less as compared to that in the pneumococcus-negative group. These results were consistent with variations in levels of Foxp3 mRNA and retinoic acid receptor-related orphan receptor-γt mRNA in adenoidal mononuclear cells. Levels of IL-17A and IL-6 in adenoid tissue were higher in the pneumococcus-negative group, and the levels of TGF-β in adenoid tissue were lower in the pneumococcus-negative group compared to the pneumococcus-positive group. Pneumococcal carriage in children was closely associated with the expressions of Foxp3+ Treg and Th17 cells in the adenoid., Conclusion: Upregulation of Foxp3+ Treg cells might downregulate the production of Th17 cells in the adenoid, resulting in decreased scavenging of Streptococcus pneumoniae and chronic pneumococcal carriage.

Introduction: Otitis media and rhinosinusitis are commonly encountered illnesses in pediatric population. Literature reports have documented the association between the occurrence of these two conditions and even their almost identical microbiological findings. Until recently, the key factor in the association of these two conditions was considered to be the hypertrophic adenoid tissue, but within the past few years there have been evidences in the literature about the presence of bacterial biofilms on the adenoids suggesting biofilms to be also responsible for both conditions, chronic otitis media with effusion and chronic rhinosinusitis., Aim: The aim of this study was to make a microbiological analysis of the adenoid tissue specimens taken from patients with chronic otitis media with effusion and chronic or recurrent rhinosinusitis and to determine their potential for biofilms formation., Methods: After the surgical intervention, adenoidectomy, microbiological evaluation and analysis of the adenoid tissue specimens taken from 20 patients were made. Having in mind the disease history, chronic otitis media with effusion was diagnosed in all 20 patients and chronic rhinosinusitis in 9 patients., Results: The results obtained from the microbiological analyses showed many potentially pathogenic bacteria in the adenoids that were almost identical with the most common organisms incorporated in the etiopathogenesis of both conditions, in chronic otitis media with effusion and in chronic rhinosinusitis. In 7 (35%) patients Haemophylus influenzae was isolated, in 6 (30%) Streptococcus pneumoniae, in 4 (20%) Moraxella catаrrhalis, in 2 (10%) patients Staphylococcus aureus and in 1 (5%) patient Streptococcus pyogenes - group A was isolated. One bacterium was isolated from all adenoid vegetations, except in one case when two bacteria (Haemophylus influenzae and Staphylococcus aureus) were concurrently isolated., Conclusion: Our results have shown that the key role in adenoid vegetations in chronic otitis media with effusion and chronic rhinosinusitis is not only the mechanism of rhinopharyngeal obstruction, but also the presence of bacterial strains with a large potential for formation of biofilms adhered to their surface, especially in cases with symptoms of chronic otitis media with effusion and chronic rhinosinusitis that were resistant to antibiotic therapy.

Objectives: The aim of this pilot study was to investigate an association between laryngopharyngeal reflux detected by combined multiple intraluminal impedance and pH monitoring and Helicobacter pylori in adenoid hyperplasia detected with real time polymerase chain reaction (PCR)., Methods: The study group consisted of 30 children (median age 5.34 years) with extraesophageal symptoms of gastroesophageal reflux disease with adenoid hyperplasia. All children underwent adenoidectomy with subsequent PCR detection of H. pylori DNA in the tissue and multiple intraluminal impedance and pH monitoring. The most proximal impedance sensor was located 1cm caudal to the entrance of the oesophagus., Results: We found significant differences in the number of reflux episodes among patients with PCR positivity (median 35) and negativity (median 0) of H. pylori (p-value of Mann-Whitney U-test 0.0056). Patients with PCR positivity of H. pylori had significantly more reflux episodes reaching the upper oesophageal sphincter (p-value of Mann-Whitney U-test 0.023). The absence of reflux episode was the only independent factor for PCR negativity of H. pylori in the multiple logistic regression model., Conclusions: These results support the hypothesis that reflux episodes reaching the upper oesophageal sphincter may play an important role in the transmission of H. pylori into lymphoid tissue of the nasopharynx and thus may contribute to adenoid hyperplasia in children., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Objective: The ability to occur in a persistent form and to inhibit apoptosis in host cells are the features of Chlamydia pneumoniae which may be associated with adenoid hypertrophy. This study aimed to (1) demonstrate the presence of C. pneumoniae DNA in the adenoid in children qualified for adenoidectomy, (2) evaluate the correlation between the presence of C. pneumoniae in the adenoid and the child's age, the size of adenoid, the incidence of recurrent respiratory tract infections, (3) demonstrate the co-occurrence of C. pneumoniae and typical bacteria in the adenoid tissue., Methods: A group of 200 children aged from 2 to 16 years (mean age 6.4) undergoing adenoidectomy from February 2010 to May 2011 were enrolled to the study. Adenoid tissue removed during planned adenoidectomy was analyzed for the presence of C. pneumoniae by real-time PCR and for the presence of typical bacteria by bacteriological culture. Statistical analysis was conducted using SPSS PASW Statistics 18 software., Results: C. pneumoniae was detected in the adenoid in 5.5% of the 200 children. Positive results were obtained most frequently (24.1%, 7/29) in the eldest group (10-16 years). In PCR positive children the degree of choanal narrowing by an adenoid tended to be larger than in PCR negative children. The occurrence of C. pneumoniae in the adenoid was not associated with a presence of a specific typical bacterium., Conclusions: The presence of C. pneumoniae in an adenoid was most frequently detected in the children of 10-16 years qualified for adenoidectomy., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Objectives: This study aims to investigate the presence of Helicobacter pylori (H. pylori) using the real-time polymerase chain reaction (PCR) method in the adenoid tissues in children undergoing surgical operation due to adenoid hypertrophy., Patients and Methods: Adenoid tissues of 23 children (8 girls, 15 boys; mean age 6.2 years; range 3 to 9 years) who were operated with the diagnosis of adenoid hypertrophy, tonsil + adenoid hypertrophy or tonsil + adenoid hypertrophy + serous otitis media in our clinic between January 2012 and April 2012 were examined., Results: Of 23 patients, H. pylori was detected in the adenoid tissues of two (8.7%). Regurgitation was present in seven patients. However, no regurgitation was found in H. pylori-positive patients., Conclusion: In this study the presence of H. pylori in patients with adenoid hypertrophy has been demonstrated using PCR method. To be able to support the hypothesis that H. pylori has a place in etiology of adenoid hypertrophy, multicenter studies are warranted.

Helicobacter pylori (H. pylori) is a major cause of chronic gastritis and gastric ulcers and considerable evidence supports the notion that infection with this bacterium is also associated with gastric malignancy in addition to various other conditions including pulmonary, vascular and autoimmune disorders. Gastric juice infected with H. pylori might play an important role in upper respiratory tract infection. Although direct and/or indirect mechanisms might be involved in the association between H. pylori and upper respiratory tract diseases, the etiological role of H. pylori in upper respiratory tract disorders has not yet been fully elucidated. Although various studies over the past two decades have suggested a relationship between H. pylori and upper respiratory tract diseases, the findings are inconsistent. The present overview describes the outcomes of recent investigations into the impact of H. pylori on upper respiratory tract and adjacent lesions.

Monocyte/macrophage cells from human nasopharyngeal lymphoid tissue can be a source of bacteria responsible for human chronic and recurrent upper respiratory tract infection. Detection and characterization of pathogens surviving intracellularly could be a key element in bacteriological diagnosis of the infections as well as in the study on interactions between bacteria and their host. The present study was undertaken to assess the possibility of isolation of viable bacteria from the cells expressing monocyte/macrophage marker CD14 in nasopharyngeal lymphoid tissue. Overall, 74 adenotonsillectomy specimens (adenoids and tonsils) from 37 children with adenoid hypertrophy and recurrent infections as well as 15 specimens from nine children with adenoid hypertrophy, which do not suffer from upper respiratory tract infections (the control group), were studied. The suitability of immunomagnetic separation for extraction of CD14(+) cells from lymphoid tissue and for further isolation of the intracellular pathogens has been shown. The coexistence of living pathogens including Haemophilus influenzae, Staphylococcus aureus, and Streptococcus pyogenes with the bacteria representing normal nasopharyngeal microbiota inside CD14(+) cells was demonstrated. Twenty-four strains of these pathogens from 32.4 % of the lysates of CD14(+) cells were isolated. Concurrently, the fluorescent in situ hybridization (FISH) with a universal EUB388, and the species-specific probes demonstrated twice more often the persistence of these bacterial species in the lysates of CD14(+) cells than conventional culture. Although the FISH technique appears to be more sensitive than traditional culture in the intracellular bacteria identification, the doubts on whether the bacteria are alive, and therefore, pathogenic would still exist without the strain cultivation.

Conclusion: Our results encourage the notion that the pharynx could be an extragastric reservoir of Helicobacter pylori (HP). The study confirmed the presence of HP in adenotonsillar tissue in children. It could have importance in the pathophysiology of upper respiratory diseases. However, its precise role in these processes remains unclear and requires further studies., Objective: A prospective study was carried out to evaluate the presence of HP in tonsillar and adenoid tissue in children. The study focused on real-time PCR analyzing CagA and VacA genotypes of HP strains., Methods: A total of 37 consecutive pediatric patients with adenotonsillar hypertrophy indicated for surgery were observed in a prospective study. Adenoidectomy and/or tonsillectomy was performed in each patient; 49 specimens were taken, 32 from adenoids and 17 from tonsils. The presence of HP and its genotype were tested in all samples by real-time PCR analysis., Results: Of 49 samples analyzed, 48 were positive for the presence of HP (98%), so only 1 sample was negative. While the genotype VacAs1bm2 was definitely dominant in adenoid tissue, wider distribution was observed in tonsillar tissue. Cag(+) strains represented one-fifth of all samples (21%).

Background: Otitis media with effusion (OME) is a common disease among children, in the pathogenesis of which bacterial infections play a critical role. It was suggested that adenoid tissue could serve as a reservoir for bacterial infection, the eustachian tubes being the migration routes of bacteria into the middle ear cavity. The aim of this study was to investigate the genetic similarity between isolates of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, obtained from adenoid tissue and middle ear fluid., Methods: A total of 60 specimens of middle ear fluids (MEFs) and 45 specimens of adenoid tissue were obtained from 45 children with OME. All the samples were inoculated on culture media for bacterial isolation and identification. The genetic similarity between bacterial isolates was determined by pulsed-field gel electrophoresis (PFGE)., Results: The same bacterial species were simultaneously isolated from adenoid tissue and MEFs of 14 patients, among which, 6 pairs of M. catarrhalis, 5 pairs of S. pneumoniae and 3 pairs of H. influenzae were identified., Conclusions: Based on the genetic similarities between isolate pairs, found by PFGE analysis, this study suggested that M. catarrhalis, S. pneumoniae and H. influenzae colonize the adenoid tissue, then migrate to the middle ear cavity and, hence, contribute to the total pathogenesis of OME., (Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.)