Bacteria In Chronic Rhinosinusitis Research Articles

Bacteriology of Different Phenotypes of Chronic Rhinosinusitis.

Chronic rhinosinusitis (CRS) reduces the health-related quality of life and subsequently causes a tremendous socio-economic impact. Although many studies have been conducted, few have identified a relationship between bacteriological characteristics and different phenotypes or endotypes. Therefore, this study aimed to elucidate the recent trends in bacterial cultures from different types of CRS in the Asian population. This retrospective case-control study recruited patients diagnosed with CRS who underwent functional endoscopic sinus surgery (FESS) at a tertiary hospital in Taiwan. The patients were classified into those with chronic rhinosinusitis with nasal polyps (CRSwNP)/chronic rhinosinusitis without nasal polyps (CRSsNP), eosinophilic chronic rhinosinusitis (eCRS)/non-eosinophilic chronic rhinosinusitis (NECRS), and central compartment atopic disease (CCAD)/lateral-dominant nasal polyp (LDNP) groups. The demographic data and bacteriological characteristics of the groups were analyzed. We included 503 patients, identifying no significant difference between CRSwNP and CRSsNP for several common bacteria in CRS. The number of Staphylococcus epidermidis isolates in culture was significantly higher in the NECRS group (50.46% vs. 32.56%, p = 0.0003) than that in the eCRS group. The number of methicillin-resistant Staphylococcus aureus (MRSA; 8.51% vs. 2.35%, p = 0.0221) positive isolates was significantly higher in the CCAD group than that in the LDNP group. This was the first study in Asia to analyze the relationship between bacteriological characteristics and CCAD. MRSA is significantly higher in the CCAD group than that in the LDNP group. Recognizing the unique microbiology of CRSwNP, eCRS, and CCAD is crucial when selecting antimicrobial therapy to lessen the socio-economic impact. 3 Laryngoscope, 134:1071-1076, 2024.

S-Nitrosoglutathione Reduces the Density of Staphylococcus aureus Biofilms Established on Human Airway Epithelial Cells.

Patients with chronic rhinosinusitis (CRS) often show persistent colonization by bacteria in the form of biofilms which are resistant to antibiotic treatment. One of the most commonly isolated bacteria in CRS is Staphylococcus aureus (S. aureus). Nitric oxide (NO) is a potent antimicrobial agent and disperses biofilms efficiently. We hypothesized that S-nitrosoglutathione (GSNO), an endogenous NO carrier/donor, synergizes with gentamicin to disperse and reduce the bacterial biofilm density. We prepared GSNO formulations which are stable up to 12 months at room temperature and show the maximum amount of NO release within 1 h. We examined the effects of this GSNO formulation on the S. aureus biofilm established on the apical surface of the mucociliary-differentiated airway epithelial cell cultures regenerated from airway basal (stem) cells from cystic fibrosis (CF) and CRS patients. We demonstrate that for CF cells, which are defective in producing NO, treatment with GSNO at 100 μM increased the NO levels on the apical surface and reduced the biofilm bacterial density by 2 log units without stimulating pro-inflammatory effects or inducing epithelial cell death. In combination with gentamicin, GSNO further enhanced the killing of biofilm bacteria. Compared to placebo, GSNO significantly increased the ciliary beat frequency (CBF) in both infected and uninfected CF cell cultures. The combination of GSNO and gentamicin also reduced the bacterial density of biofilms grown on sinonasal epithelial cells from CRS patients and improved the CBF. These findings demonstrate that GSNO in combination with gentamicin may effectively reduce the density of biofilm bacteria in CRS patients. GSNO treatment may also enhance the mucociliary clearance by improving the CBF.

The role of TAS2R38 genotype in surgical outcomes and culturable bacteria in chronic rhinosinusitis with or without nasal polyps.

Recent studies reported the relationship between genetic variations and TAS2R38, which is a bitter taste receptor expressed in the cilia of human sinonasal epithelial cells, among the predisposing factors playing role in immune response to upper respiratory tract bacterial infection. The present study aims to examine the relationship of TAS2R38 genotype with the active microorganism and the effect of genotype on the surgical outcomes among chronic rhinosinusitis patients. 34 patients undergoing endoscopic sinus surgery (ESS) for chronic rhinosinusitis with or without polyps (23 CRSwNP, 11 CRSsNP) and 30 patients undergoing septoplasty surgery for isolated nasal septum deviation were included. All the patients were genotyped for TAS2R38. Scoring was made using endoscopic Modified Lund-Kennedy and radiological Lund-Mackay systems preoperatively. Sino-Nasal Outcome Test with 22 items (SNOT-22) was implemented preoperatively and postoperatively. Nasal swab culture samples were taken intraoperatively from CRS patients and the active microorganism were isolated. In the TAS2R38 genotyping of the study group, PAV/PAV was found in 32.4% of patients, PAV/AVI in 47.1%, and AVI/AVI in 20.6%. In the control group, PAV/PAV was found in 26.7%, PAV/AVI in 36.7%, and AVI/AVI in 36.7%. In the study group, there was no statistically significant difference between the CRS and CRS subgroups in terms of TAS2R38 genotype distributions. The changes in patients' preoperative and postoperative SNOT-22 scores were similar between the genotypes. Proliferation was detected in culture in the whole AVI-AVI group, 81.8% of PAV-PAV group, and 56.3% of PAV-AVI group but the difference was not found to be statistically significant. The proliferation level of Staphylococcus epidermidis by TAS2R38 genotype was found to be statistically significantly higher among patients, who had AVI-AVI genotype, in CRSwNP. We did not find a statistically significant relationship between the TAS2R38 genotype and CRS subtype, sinonasal bacterial infection risk increase and surgical success rate in CRS patients. Long-term and large-scale studies are needed, which are to be carried out by individual genotyping and sequencing to provide more information on the effects of these genetic variants.

Association between mucosal barrier disruption by Pseudomonas aeruginosa exoproteins and asthma in patients with chronic rhinosinusitis.

Chronic rhinosinusitis (CRS) is a common chronic respiratory condition, frequently associated with asthma and affecting the majority of cystic fibrosis (CF) patients. Pseudomonas aeruginosa infections and biofilms have been implicated in recalcitrant CRS. One of the mechanisms of action for bacteria in CRS and CF is mucosal barrier disruption by secreted products that contribute to the inflammation. However, the role of biofilm and planktonic forms of P. aeruginosa in this process is not known. The aim is to determine the effect of P. aeruginosa exoproteins isolated from CF and non-CF CRS patients on the mucosal barrier. Exoproteins from 40 P. aeruginosa isolates were collected in planktonic and biofilm forms and applied to air-liquid interface (ALI) cultures of primary human nasal epithelial cells (HNECs). Mucosal barrier integrity was evaluated by transepithelial electrical resistance (TEER), passage of FITC-dextrans and immunofluorescence of tight junction proteins. Cytotoxicity assays were performed to measure cell viability, and IL-6 ELISA was carried out to evaluate pro-inflammatory effects. Planktonic exoproteins from 20/40 (50%) clinical isolates had a significant detrimental effect on the barrier and significantly increased IL-6 production. Barrier disruption was characterized by a reduced TEER, increased permeability of FITC-dextrans and discontinuous immunolocalization of tight junction proteins and was significantly more prevalent in isolates harvested from patients with comorbid asthma (P<.05). Exoproteins from planktonic P. aeruginosa clinical isolates from asthmatic CRS patients have detrimental effects on the mucosal barrier and induce IL-6 production potentially contributing to the mucosal inflammation in CRS patients.

Diverse phenotypes and endotypes of fungus balls caused by mixed bacterial colonization in chronic rhinosinusitis.

The pathogenic roles of fungus and bacteria in chronic rhinosinusitis (CRS) remain unclear. Recently, we described the bacterial ball, which is distinct from the fungus ball, as an unusual phenotype of bacterial infection. In this study, we investigated the clinical, histopathologic, and immunologic characteristics of sinonasal microorganic materials, including fungus ball and bacterial ball. In this study, we enrolled 80 CRS patients with sinonasal microorganic materials and 10 control subjects who underwent skull base surgery or endoscopic dacryocystorhinostomy and had no signs or symptoms of nasal inflammation. All specimens were stained with hematoxylin-eosin, Gomori-methenamine-silver, and Gram stain to identify fungal organisms and Gram-positive/negative bacterial colonies. The expression of tumor necrosis factor (TNF)-α; interleukin (IL)-1β; S100A7; S100A8/A9; and short, palate, lung, and nasal epithelial clone 1 (SPLUNC1) were evaluated by enzyme-linked immunosorbent assay using sinus lavage fluid. We histologically classified sinonasal microorganic materials into the following 4 groups: fungus ball (n = 45); bacterial ball (n = 6); mixed ball (formed by a mixture of fungus and bacteria, n = 27); and double ball (formed by separate fungal and bacterial balls, n = 2). Compared with the fungus ball, the mixed ball was more frequently detected in immunocompromised patients (p < 0.0001). In addition, TNF-α expression was significantly higher in fungus and mixed balls than in control, whereas the mixed ball showed higher expression of IL-1β compared with the fungus ball. Moreover, the expression of S100A7 and S100A8/A9 protein in the mixed ball was significantly decreased when compared with the fungus ball, whereas there was no significant difference in SPLUNC1 expression between fungus and mixed balls. Our findings suggest that fungal and bacterial interactions are diverse in CRS. Specifically, the mixed ball is prevalent in CRS with an immunocompromised state and it may decrease epithelial barrier function.

Microbiome and Culture Based Analysis of Chronic Rhinosinusitis Compared to Healthy Sinus Mucosa.

The role of bacteria in chronic rhinosinusitis (CRS) is still not well understood. Whole microbiome analysis adds new aspects to our current understanding that is mainly based on isolated bacteria. It is still unclear how the results of microbiome analysis and the classical culture based approaches interrelate. To address this, middle meatus swabs and tissue samples were obtained during sinus surgery in 5 patients with CRS with nasal polyps (CRSwNP), 5 patients with diffuse CRS without nasal polyps (CRSsNP), 5 patients with unilateral purulent maxillary CRS (upm CRS) and 3 patients with healthy sinus mucosa. Swabs were cultured, and associated bacteria were identified. Additionally, parts of each tissue sample also underwent culture approaches, and in parallel DNA was extracted for 16S rRNA gene amplicon-based microbiome analysis. From tissue samples 4.2 ± 1.2 distinct species per patient were cultured, from swabs 5.4 ± 1.6. The most frequently cultured species from the swabs were Propionibacterium acnes, Staphylococcus epidermidis, Corynebacterium spp. and Staphylococcus aureus. The 16S-RNA gene analysis revealed no clear differentiation of the bacterial community of healthy compared to CRS samples of unilateral purulent maxillary CRS and CRSwNP. However, the bacterial community of CRSsNP differed significantly from the healthy controls. In the CRSsNP samples Flavobacterium, Pseudomonas, Pedobacter, Porphyromonas, Stenotrophomonas, and Brevundimonas were significantly enriched compared to the healthy controls. Species isolated from culture did not generally correspond with the most abundant genera in microbiome analysis. Only Fusobacteria, Parvimonas, and Prevotella found in 2 unilateral purulent maxillary CRS samples by the cultivation dependent approach were also found in the cultivation independent approach in high abundance, suggesting a classic infectious pathogenesis of odontogenic origin in these two specific cases. Alterations of the bacterial community might be a more crucial factor for the development of CRSsNP compared to CRSwNP. Further studies are needed to investigate the relation between bacterial community characteristics and the development of CRSsNP.

The impact of bitter taste receptor genetics on culturable bacteria in chronic rhinosinusitis.

Extra-oral bitter taste receptors have been associated with innate bacterial defence mechanisms. Genetic variation in T2R38 functionality has been shown to be associated with susceptibility to upper respiratory tract infections and chronic rhinosinusitis (CRS). We sought to independently assess the influence of bitter taste receptor genotype on the presence of culturable bacteria in the sinuses. A cross-sectional analysis of patients with CRS undergoing surgery was performed. Middle meatal nasal swabs were sent for microbiological evaluation at the time of the procedure. Mucosal biopsies were taken and sent for bitter taste receptor genotype analysis. Sequencing of 3 polymorphisms in the TAS2R38 gene was performed to identify genotypes as super-tasters (PAV/PAV), non-tasters (AVI/AVI) or heterozygous expression (PAV/AVI). The presence of culturable organisms and common pathogens were compared with bitter taste receptor genotypes. 25 patients (age 52.4 +/- 18.28 years, 51% female) were assessed. Super-tasters comprised 16% of the group, 24% were non-tasters and 48% had heterozygous expression. A cultured pathogen was grown in 48% of patients; 32% gram-positive, 20% gram-negative, 28% grew Staphylococcus aureus and 12% Pseudomonas aeruginosa. A non-taster genotype was predictive of colonised pathogens. Tissue eosinophilia (more than 10 HPF) was seen in 48%. Even in a small sample of patients with CRS, non-taster T2R38 genotype appears to predict the presence of culturable bacteria colonising the sinus cavity at the time of surgery for their condition. A genetic link to patients more likely to become infected is likely.

The impact of bitter taste receptor genetics on culturable bacteria in chronic rhinosinusitis

The impact of bitter taste receptor genetics on culturable bacteria in chronic rhinosinusitis

Bacterial community collapse: a meta-analysis of the sinonasal microbiota in chronic rhinosinusitis.

Chronic rhinosinusitis (CRS) is a common, debilitating condition characterized by long-term inflammation of the nasal cavity and paranasal sinuses. The role of the sinonasal bacteria in CRS is unclear. We conducted a meta-analysis combining and reanalysing published bacterial 16S rRNA sequence data to explore differences in sinonasal bacterial community composition and predicted function between healthy and CRS affected subjects. The results identify the most abundant bacteria across all subjects as Staphylococcus, Propionibacterium, Corynebacterium, Streptococcus and an unclassified lineage of Actinobacteria. The meta-analysis results suggest that the bacterial community associated with CRS patients is dysbiotic and ecological networks fostering healthy communities are fragmented. Increased dispersion of bacterial communities, significantly lower bacterial diversity, and increased abundance of members of the genus Corynebacterium are associated with CRS. Increased relative abundance and diversity of other members belonging to the phylum Actinobacteria and members from the genera Propionibacterium differentiated healthy sinuses from those that were chronically inflamed. Removal of Burkholderia and Propionibacterium phylotypes from the healthy community dataset was correlated with a significant increase in network fragmentation. This meta-analysis highlights the potential importance of the genera Burkholderia and Propionibacterium as gatekeepers, whose presence may be important in maintaining a stable sinonasal bacterial community.

Bacteriology of the Middle Meatus in Chronic Rhinosinusitis with and without Polyposis

There are only limited data in the literature, and none specifically from the Middle East, on the pathogenic bacteria in chronic rhinosinusitis (CRS) as opposed to healthy nasal cavities and their association with disease severity. The present study was conducted in the department of head and neck surgery of a tertiary medical center. Middle meatal swabs were taken preoperatively from patients with CRS with nasal polyposis (CRSwNP) (n = 60), CRS without nasal polyposis (CRSsNP) (n = 50), and control patients with septal deviation (n = 26) or no nasal abnormalities (n = 27). Culture findings were compared among the groups and correlated with CRS severity. Positive pathogenic culture rates were 78% in the CRSwNP group and 64% in the CRSsNP group. Twenty pathogenic bacterial species were identified; the most common was Staphylococcus aureus (27%). The most common Gram-negative isolate was Citrobacter spp. (17%). Gram-negative species were significantly more prevalent in the CRSwNP group than the others. Mean Lund-Mackay scores were 12.8 in the CRSwNP group and 6.9 in the CRSsNP group, and were unrelated to the culture findings. Positive culture rates were significantly higher in the septal deviation (54%) than the nasal healthy group (26%), although both values were significantly lower than in the CRS groups. In conclusion, patients with CRS have higher rates of bacterial isolates than patients without CRS. CRSwNP is associated with more Gram-negative bacteria than CRSsNP, regardless of disease severity. The relatively high positive culture rate in patients with septal deviation merits investigation.

The characteristics of intramucosal bacteria in chronic rhinosinusitis: a prospective cross-sectional analysis

We have observed subepithelial bacterial microcolonies within the mucosa of patients with chronic rhinosinusitis (CRS). These were predominantly Staphylococcus aureus and did not appear to elicit a local inflammatory response. We hypothesized that these microcolonies had made adaptations allowing them to exist apparently undetected within the mucosa. We sought to determine whether the tissue colonies had genotypic or phenotypic variations from the surface bacteria. Mucosal swabs and tissue biopsies were taken from 31 patients with CRS undergoing functional endoscopic sinus surgery, and 9 with normal sinuses having transnasal pituitary surgery. Biopsied tissues were assessed histologically, by routine culture, and by culture techniques facilitating growth of small colony variants (SCVs). Genotypic typing compared isolates of S. aureus that were cultured from both swab and tissue samples. The activity of the accessory gene regulator (agr) gene, a global regulator of S. aureus virulence, was evaluated indirectly by determining the hemolytic activity of the colonies on blood agar. SCVs were grown from 2 samples but these were found not to possess the nuc gene, specific to S. aureus. When S. aureus was recovered from both swab and mucosa, the genetic profiles were indistinguishable in all but 1 patient. All S. aureus cultured from mucosa demonstrated β-hemolysis, implying normal agr activity. Intramucosal S. aureus are genetically closely related and phenotypically similar to surface S. aureus. Further studies are needed to explore the possible mechanisms by which intramucosal colonies become less immunogenic, and the role of the colonies in the pathophysiology of CRS.

Intramucosal Bacterial Microcolonies Exist in Chronic Rhinosinusitis without Inducing a Local Immune Response

Although chronic rhinosinusitis (CRS) causes very significant morbidity, much about its pathogenesis remains uncertain. Recent studies have identified polymicrobial biofilms on the surface of sinus mucosa and Staphylococcus aureus within the sinus mucosa of patients with CRS, both with and without nasal polyps. The pathogenic implications of intramucosal bacteria in CRS are unknown. This study was designed to determine the prevalence and species of bacterial colonies within the sinus mucosa of adult patients with and without CRS and to describe the relationship of these bacterial colonies to the host immune response. Sinus mucosa from patients with and without CRS was examined using Gram and Giemsa staining, immunohistochemistry, bacterial culture, and fluorescence in situ hybridization techniques. Bacterial microcolonies were observed within the mucosa in 14 of 18 patients with CRS. In 10 of these patients colonies were positively identified as S. aureus. Staphylococcal microcolonies were observed at a lower level (1 of 8 patients) in normal sinus mucosa. There was no correlation between detection of S. aureus on the mucosal surface and microcolonization of the mucosa. Surprisingly, there was no evidence of an immune reaction to microcolonies. Indeed, fewer T lymphocytes (p = 0.03) and eosinophils (p = 0.03) were counted immediately surrounding the microcolonies compared with uninfected areas of the same tissue. Bacterial microcolonies are prevalent within paranasal sinus mucosa and are commonly S. aureus. These microcolonies do not provoke immune detection and may represent a phenotype that actively evades host immunity. This may underpin the recalcitrance of CRS to antibiotic therapy. These findings challenge classic views of both infection and mucosal immunity in human chronic disease. The presence of intramucosal bacteria in samples of normal sinus mucosa also questions the sensitivity of detecting nasal carriage of pathogens by swabbing the surface of the anterior nares.

Chronic rhinosinusitis in children—Bacteriological analysis in terms of cytological examination

IntroductionThe mechanisms of inflammatory response occurring in chronic rhinosinusitis in children are multifactorial. Besides the history and a physical examination, amongst diagnostic tools there are cytological and bacteriological examinations. Objectives1Determining the nature of the bacterial flora present in the nasal cavities and paranasal sinuses in children with chronic rhinosinusitis amongst patients of The Department of Paediatric Otolaryngology, Warsaw Medical University.2Determining the relation between bacterial strains and cytological examination of nasal mucosa in children with chronic rhinosinusitis. Materials and methodsThe study group included 64 patients with chronic rhinosinusitis without polyps. The control group included 30 randomly chosen children. Diagnostic tests performed in both groups were: middle meatal culture and cytological examination from the inferior nasal concha and middle meatus. Statistical analysis was accomplished with Statistica 8.0. Results1The most common strains of bacteria found in patients with chronic rhinosinusitis are gram-positive bacteria such as coagulase-negative staphylococci and Staphylococcus aureus, and gram-negative bacteria such as: Haemophilus influenzae and Moraxella catarrhalis.2The analysis of relation between bacterial strains and nasal cytology revealed:(a)The presence of the coagulase-negative staphylococci was correlated with less severe inflammatory changes, expressed by a higher count of epithelial columnar cells and lower count of neutrophils in the cytology results.(b)The presence of S. aureus was associated with a significantly higher percentage of squamous cells in cytology obtained from the middle nasal turbinate (for S. aureus found in the maxillary sinus). This observation suggests drawing attention to the transformation of mucosa into squamous, as a possible risk factor for colonisation of S. aureus in the maxillary sinus.(c)Occurrence of H. influenzae and M. catarrhalis in the smear correlated with a greater potential for infection, expressed by a significantly higher count of neutrophils. ConclusionsDamage to the respiratory epithelial surface is understood as damage to the innate immune barrier, and repeated antibiotic therapy with the subsequent repopulation of the epithelium accidentally by various bacteria can become responsible for the pathogenic effect of bacteria in chronic rhinosinusitis.

Current issues in the management of sinonasal methicillin-resistant Staphylococcus aureus

Staphylococcus aureus is an important pathogen, contributing to both pediatric and adult infections in the USA. In recent years, methicillin-resistant Staphylococcus aureus (MRSA) has become a source of public fear and outcry. In this article, we review facts and fiction of sinonasal MRSA. Although the exact role of bacteria in chronic rhinosinusitis continues to be debated, the role of MRSA in these patients is even less apparent. When MRSA is involved, there are few to guide treatment. Oral, parenteral, and topical antibiotics are all used in the treatment of sinonasal MRSA. MRSA eradication has been advocated by some; however, there are few data to support the efficacy of eradication. Current treatment regimens by the authors are reviewed. There is much that is not known about the role of MRSA in sinonasal infection, and the impact of sinonasal MRSA on the health quality of the community also remains undefined. Treatment outlines are presented; however, with the current state of data, these treatment preferences lack a solid, evidence-based foundation that is favored. Future studies will lay the foundation for more rigorously supported treatment algorithms.

Bacterial Cultures of the Middle Meatus and Bronchoalveolar Lavage in Chronic Rhinosinusitis

The role of bacteria in chronic rhinosinusitis (CRS) is still debated and their effect on the lower airways is less known. The aim of this study was to identify the individual bacterial species in CRS and to correlate these microbiological data with the clinical findings and associated bacterial cultures of the lower airways. This prospective study included 25 CRS patients scheduled for functional endoscopic sinus surgery. Preoperatively, pulmonary function and CT examinations were performed together with endoscopic and clinical assessment. Under aseptic conditions, with the patient under general anesthesia, 25 bronchoalveolar lavages (BALs) via the endotracheal tube and 50 endoscopically guided middle meatal specimens were obtained. All the specimens were examined for aerobic and anaerobic bacteria and correlated to the associated neutrophilic cellular changes. Positive bacterial cultures were obtained in 71% of the middle meatal specimens and 8% of the BAL specimens. Three subgroups of lower airway involvement were identified, i.e. asthma, small airway disease (SAD) and normal lower airways. In SAD only, there was a significant correlation between the cultured bacteria of the left (p = 0.04) and the right (p = 0.04) middle meatus and the presence of local neutrophilia. In asthma, middle meatal bacterial cultures correlated with nasal blockage (p = 0.04) and CT Lund-Mackay score (p = 0.04). Conclusion: Middle meatal cultures were frequently positive, mainly aerobic Gram-positive bacteria. In SAD, a mixed flora of Gram-positive aerobic bacteria and anaerobic bacteria was observed that correlated with neutrophilic inflammation. BAL cultures do not show infection or colonization of the lower airways, despite functional abnormalities in 60% of the patients.

Use of an in Vitro Assay for Determination of Biofilm-Forming Capacity of Bacteria in Chronic Rhinosinusitis

Bacterial biofilms are increasingly implicated in the pathogenesis of chronic disease and have been established in several chronic ear, nose, and throat conditions, including chronic sinusitis (CRS). However, this relies on specialized imaging methods not widely available. We wished to assess the capacity of an easily performed, inexpensive in vitro test to assess biofilm production by bacteria recovered from individuals with CRS with or without nasal polyposis. Bacterial isolates were recovered from patients consulting an academic tertiary rhinology practice. Biofilm formation was determined with an in vitro staining method using crystal violet. Ten isolates of Pseudomonas aeruginosa, Staphylococcus aureus, and 11 of coagulase-negative staphylococcus from patients with CRS having previously undergone endoscopic sinus surgery for >1 year were assessed. Samples were cultured 24 hours at 37 degrees C on 96-well plates in tryptic soy broth 0.5% glucose medium. After staining with crystal violet, optical density at 570 nm was measured to quantify biofilm production. Biofilm-forming capacity was compared with positive and negative controls for each species obtained from commercial sources. Positive controls all grew biofilms, with a tendency of lesser biofilm formation at higher dilutions. Twenty-two of 31 clinical samples produced a biofilm greater or equal to the positive control. Biofilm was recovered consistently for all three species studied. This in vitro assessment method is capable of detecting biofilm-forming capacity in bacteria recovered from individuals with CRS. This simple assay may be a useful complement to existing techniques for clinical research.

Bacterial Infection in Chronic Rhinosinusitis: A Controlled Paired Analysis

The aim of this study was to determine if pathogenic bacteria are involved in the pathogenesis of chronic rhinosinusitis (CRS). A consecutive series of adult patients with unilateral sinus disease determined by unilateral radiographic involvement or unilateral purulent secretions was microbiologically studied. Aerobic and anaerobic bacterial and fungal cultures were obtained during endoscopic sinus surgery from purulent secretions or tissue culture. Positive culture rates were compared between the diseased sinus and the contralateral nondiseased (control) sinus to determine if pathogenic bacteria were more commonly recovered from the diseased sinuses. Forty-nine adult patients completed the study with appropriate microbiological data. Coagulase-negative staphylococci were the most commonly recovered bacteria followed by Staphylococcus aureus from the diseased side of the sinuses with similar findings for the control sinus. Bacterial species were recovered from 87.8% of the diseased side of the sinuses versus 85.7% from the control sinuses (p = 0.50). Reanalysis with coagulase-negative staphylococci considered as nonpathogen showed a 46.9 and 49.0% positive bacterial culture rate in diseased and control groups, respectively (p = 0.50). No significant difference in positive anaerobic culture rates were identified between groups (59.1% diseased versus 55.1% control, respectively, p = 0.61). Antibiotic resistance rates were no different between bacteria cultured from diseased sinuses versus control (p = 0.115). Both aerobic and anaerobic bacterial species may be recovered from both diseased and nondiseased sinuses in patients with CRS. These findings cast some doubt on the exact etiologic role of bacteria in CRS, suggesting other factors or other agents also may be responsible in CRS pathogenesis.

The Role of Atypical Bacteria in Chronic Rhinosinusitis

This study examines the presence of atypical bacteria in chronic rhinosinusitis (CRS) by utilizing the polymerase chain reaction (PCR). Eleven patients with CRS were prospectively enrolled. DNA was isolated from the mucosa samples and subjected to PCR using oligonucleotides specific for identification of atypical bacteria including: Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella pneumophila. Also, routine aerobic and anerobic cultures were processed. Sinus tissue samples from 6 fresh corpses without evident sinus disease served as controls. No atypical bacteria were identified in the 11 patient samples by PCR. One of 6 cadaver controls was positive by PCR for M pneumoniae. The most common organism obtained from routine culture was coagulase negative staphylococci. Using sensitive PCR techniques, CRS mucosa did not reveal DNA from M pneumoniae, C pneumoniae, or L pneumophila. Atypical bacteria were not identified in patients with CRS despite highly effective PCR methods and they may not play a significant role in the cause of CRS.

The role of atypical bacteria in chronic rhinosinusitis

Problem: Conflicting studies exist on the microbiology of chronic rhinosinusitis (CRS), but none identify atypical bacteria, owing to these bacteria’s fastidious nature. Atypical bacteria are common causes of pneumonia. However, their role in upper respiratory disease is less clear. Recently the polymerase chain reaction has been valuable in detecting aerobic and anaerobic bacteria, fungi, and viruses implicated in chronic rhinosinusitis. This study examines the role of atypical bacterial infection in CRS by utilizing the polymerase chain reaction. Methods: Eleven patients (8 males, 3 females) with CRS were prospectively enrolled. All failed medical therapy requiring endoscopic sinus surgery (ESS). Ethmoid sinus mucosa was processed by polymerase chain reaction (PCR) to identify the presence of Mycoplasma pneumoniae, Chlamydia pneumoniae, and Legionella pneumophila. Also, intra-operative routine aerobic and anaerobic cultures were processed. The control group included 6 fresh corpses (3 males, 3 females) without evident sinus disease from the Miami-Dade County Medical Examiner. Their ethmoid mucosa was also obtained endoscopically and processed by PCR. Results: Of the 17 ethmoid mucosa samples (11 patients, 6 cadaver controls), 1 cadaver control was positive by PCR for M pneumoniae. All other samples were negative for M pneumoniae, C pneumoniae, and L pneumophila. The most common organism obtained from routine culture was coagulase negative staphylococci. Conclusion: Using highly sensitive PCR techniques, chronic rhinosinusitis mucosa did not reveal DNA from M pneumoniae, C pneumoniae, or L pneumophila. Atypical bacterial infection may not play a significant role in adult CRS. Significance: This study demonstrates that PCR is highly sensitive in detecting atypical bacteria and these bacteria are not identified in patients with CRS. Support: None reported.