Biofilms In Patients Research Articles

Comparative evaluation of changes in the periodontal microbiome in patients with chronic generalized periodontitis after Vector-therapy

Relevance. The development and implementation into wide clinical practice an effective methods for removing subgingival biofilms, which allow qualitatively removing periodontopathogens and preserving the structure of the root surface for the subsequent restoration of the supporting-retaining apparatus of the tooth, is an important aspect in the treatment of chronic generalized periodontitis. Purpose. To evaluate the effectiveness of removing microbial biofilm in patients with chronic generalized periodontitis using Vector-therapy in comparison with hand-held instruments. Materials and methods . The study involved 119 patients (68 women and 51 men) in the age of 26-70 years (mean age 47.0 ± 12.5 years) with chronic generalized periodontitis of moderate and severe degree. The patients were divided into 3 groups depending on the method of root surface treatment (Curettes, Vector Paro, combined Curette Vector Paro treatment). Patients of all groups was evaluated such parameters, as BOP, PD / HPA, CAL / RFP and radiological parameters. For microbiological examination, real-time PCR was used with the sampling of the contents of the periodontal pocket before treatment for 10 days and 6 weeks. The dynamics of clinical indicators, frequency of occurrence, absolute, and the participation of key periodontal pathogenic microorganisms (A. actinomycetemcomitans, P. Gingivalis, P. intermedia, T. forsythia, T. denticola) and Candida albicans from the total bacterial mass of the contents of the periodontal pocket were assessed. The results were processed statistically using the Statistica software. Results. When comparing the groups with each other, a statistically significant relationship was found for a decrease in PB in group 2 (Vector) and group 3 (Curettes Vector Paro) in group 1 (Curettes), an inverse correlation between the level of bleeding (BOR) at a follow-up of 10 days (rs -0.463, p < 0.001) and 6 weeks (rs -0.342, p-value 0.025) and by the method of removing subgingival dental plaque. Statistically significant changes in the frequency of occurrence, number and proportion of periodontal pathogenic microorganisms in the groups were found in the dynamics of observation, indicating changes in the structure of the subgingival microbiome. Conclusion. The inclusion of Vector therapy in conservative treatment significantly accelerates the timing of inflammation in the periodontal tissues and leads to some significant shifts in the structure of the subgingival microbiome.

Analysis of subclinical infections and biofilm formation in cases of capsular contracture after silicone augmentation rhinoplasty: Prevalence and microbiological study.

BackgroundImplant-related deformities in aesthetic rhinoplasty are a major problem for rhinoplasty surgeons. Capsular contracture is believed to be the pathological cause of delayed contour deformities, comparable to breast implant-related contracture. This study investigated the prevalence of bacterial biofilms and other epidemiological factors related to capsular contracture in cases of silicone augmentation rhinoplasty.MethodsThirty-three patients who underwent corrective rhinoplasty due to a delayed contour deformity or aesthetic revision after implant rhinoplasty were studied from December 2014 to December 2016. All recruited patients received surgical correction by the authors. The patients were categorized by clinical severity into four grades. Demographic data and related confounding factors were recorded. Samples of capsular tissue and silicone removed from each patient were analyzed for the presence of a biofilm by ultrasonication with bacterial culture and scanning electron microscopy.ResultsThirty-three paired samples of capsular tissue and silicone implants from the study group were analyzed. Biofilms were detected in one of 10 subjects (10%) with grade 1 contracture, two of four (50%) with grade 2 contracture, 10 of 14 (71.40%) with grade 3 contracture, and four of five (80%) with grade 4 contracture (P<0.05). The organisms found were Staphylococcus epidermidis (47.10%), coagulase-negative staphylococci (35.30%), and Staphylococcus aureus (17.60%).ConclusionsAs with breast implant-related capsular contracture, silicone nasal augmentation deformities likely result from bacterial biofilms. We demonstrated the prevalence of biofilms in patients with various degrees of contracture. Implant type and operative technique seemed to have only vague correlations with biofilm presence.

Antibiotic Sensitivity Patterns of Urine and Biofilms in Patients with Indwelling Urinary Catheter in Denden Hospital, Asmara, Eritrea

The intricate infections leading to long-term morbidity of catheterized patients are due to the presence of a covering and blocking the lumen of urinary catheters by biofilms which have increased ability of resistance to host immune system and antibiotic treatment. The biofilm mode of growth is a basic survival strategy implemented by bacteria in a wide range of settings such as environmental, industrial and clinical aquatic settings. Bacterial growth on the inner surface of the catheter with biofilm formation is frequent and may occur within days of catheter placement. This study investigated the formation of biofilm inside catheter lumen of patients from Denden hospital, Asmara, Eritrea. And also, it assessed the antimicrobial sensitivity pattern of biofilm isolates and compared it with urine isolates. Resistance to antibiotics was observed in biofilm isolates more than urine isolates. E. coli was the most frequently isolated organism in both biofilm and urine samples.

Features of filiform papillary lesion of the tongue and its mucosal microflora biofilm in patients with gastroesophageal reflux disease

To identify filiform papillary lesions, localization changes, and the composition of the microflora of the dorsal lingual surface in patients with gastroesophageal reflux disease (GERD). Dorsal lingual surface biopsy specimens were investigated in 7 patients with GERD (3 men, 4 women) and in 6 individuals without digestive diseases (3 men, 3 women). The diagnosis of GERD was based on a set of clinical data, daily pH-metry, EGDS, and the specialized GerdQ questionnaire. Scanning electron microscopy and fluorescence confocal microscopy were used. Intact filiform lingual papillae had a complex structure and consisted of primary and secondary papillae. Foci of increased epithelial desquamation with partial or complete loss of secondary papillae were detected in patients with GERD. There was a microflora biofilm only on the epithelium of the secondary papillae in the intact areas and in the areas with preserved secondary papillae in patients with GERD. On the contrary, the foci of lesion and those with completely lost secondary papillae in GERD patients exhibited the microflora (more diverse in its morphological characteristics) on the surface of the epithelial cells of primary papillae, except for their cup-shaped hollows in the area of the lost secondary processes. These cup-shaped hollows preserved dense intercellular contacts of epithelial cells. The lingual mucosal filiform papillae in GERD patients are characterized by the appearance of areas with partial or complete loss of secondary papillae and with increased desquamation of epithelial cells. There is microflora biofilm translocation to the primary papillae, except for their cup-shaped hollows (an area of the lost secondary papillae). Translocation of the microflora increased its morphological diversity.

Effect of ozonized olive oil on oral levels of Candida spp. in patients with denture stomatitis

Objective: The aim of this study was to evaluate the effect of ozonized oil (OZ) on the oral levels of Candida spp. in patients with denture stomatitis. Material and Methods: In vitro tests were performed to validate antifungal activity and to standardize OZ conditions. Antifungal activity was screened against C. albicans and five non-albicans species (C. tropicalis, C. dubliniensis, C. krusei, C. guilliermondii, and C. parapsilosis). Also, the effects on C. albicans planktonic and biofilm were evaluated. After validation, OZ was included in a therapeutic protocol of denture stomatitis in vivo. Thirty patients used OZ and 20 used sodium bicarbonate (SB) for 14 days. After 7 and 14 days, clinical evaluation, isolation and identification of yeasts were performed. Isolates were identified by phenotypic and genotypic tests. Ozonized oil showed in vitro antifungal activity against all species of Candida. Ozonized oil reduced the number of viable cells in C. albicans biofilms. Oral candidal levels were lower in relation to baseline both after after 14 days of treatment with SB and OZ. Results: A total of 493 Candida spp. isolates was obtained and 80% were identified as C. albicans. Remission of denture stomatitis was observed in all patients after 7 days of treatment in both groups. Conclusion: Within the limits of the study we can conclude that ozonized olive oil can be a new alternative for the control of biofilm in patients with denture stomatitis. KeywordsOzone; Candida; Antifungal Agents; Stomatitis; Denture.

The proteins of Fusobacterium spp. involved in hydrogen sulfide production from L-cysteine

BackgroundHydrogen sulfide (H2S) is a toxic foul-smelling gas produced by subgingival biofilms in patients with periodontal disease and is suggested to be part of the pathogenesis of the disease. We studied the H2S-producing protein expression of bacterial strains associated with periodontal disease. Further, we examined the effect of a cysteine-rich growth environment on the synthesis of intracellular enzymes in F. nucleatum polymorphum ATCC 10953. The proteins were subjected to one-dimensional (1DE) and two-dimensional (2DE) gel electrophoresis An in-gel activity assay was used to detect the H2S-producing enzymes; Sulfide from H2S, produced by the enzymes in the gel, reacted with bismuth forming bismuth sulfide, illustrated as brown bands (1D) or spots (2D) in the gel. The discovered proteins were identified with liquid chromatography – tandem mass spectrometry (LC-MS/MS).ResultsCysteine synthase and proteins involved in the production of the coenzyme pyridoxal 5′phosphate (that catalyzes the production of H2S) were frequently found among the discovered enzymes. Interestingly, a higher expression of H2S-producing enzymes was detected from bacteria incubated without cysteine prior to the experiment.ConclusionsNumerous enzymes, identified as cysteine synthase, were involved in the production of H2S from cysteine and the expression varied among Fusobacterium spp. and strains. No enzymes were detected with the in-gel activity assay among the other periodontitis-associated bacteria tested. The expression of the H2S-producing enzymes was dependent on environmental conditions such as cysteine concentration and pH but less dependent on the presence of serum and hemin.

Factors affecting dental biofilm in patients wearing fixed orthodontic appliances

BackgroundThe aim of this study is to investigate the amount and the distribution of biofilm in patients wearing fixed appliances and its relation with age, gender, frequency of tooth brushing, and patient motivation.MethodsThe sample comprised 52 patients (15.5 ± 3.6 years old, 30 females and 22 males) wearing fixed orthodontic appliances. Dental biofilm was assessed using a modified plaque index (PI). A questionnaire was used to collect patient’s information, including gender, age, treatment motivation, and frequency of tooth brushing.ResultsGingival (PI score = 0.9 ± 0.7), mesial (0.8 ± 0.6), and distal (0.8 ± 0.5) areas accumulated more biofilm than occlusal areas (0.3 ± 0.3) (P < 0.038). The maxillary lateral incisors (1.1 ± 0.8) and maxillary canines (1.0 ± 0.8) had more biofilm than other teeth (P < 0.05). The maxillary arch (0.8 ± 0.7) had significantly more biofilm than mandibular arch (0.6 ± 0.6) (P = 0.042). No significant difference was found between the right side (0.7 ± 0.7) and left side (0.7 ± 0.6) (P = 0.627). Less biofilm was found in females (0.6 ± 0.5), adults (0.3 ± 0.3), and “self-motivated” patients (0.3 ± 0.3), compared with males (0.9 ± 0.5), children (0.8 ± 0.6), and “family-motivated” patients (1.1 ± 0.5) (P < 0.001). The amount of biofilm was associated with self-report of the frequency of daily tooth brushing (P < 0.001).ConclusionsPatients wearing fixed orthodontic appliances have the highest biofilm accumulation on the maxillary lateral incisors and maxillary canines, particularly in the gingival area and areas behind arch wires. Less biofilm was observed in female and adult patients and in those who were self-motivated and brushed their teeth more often.

Normal Skin Microbiota is Altered in Pre-clinical Hidradenitis Suppurativa.

Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease defined by recurrent nodules, tunnels (sinus tracts) and scarring involving the intertriginous regions. The clinical course of HS is compatible with a biofilm-driven disease, and biofilm has been described in lesional HS skin. We therefore hypothesized that clinically unaffected HS skin would also have an increased presence of biofilm compared with that of healthy controls. We conducted a case-control study, investigating the morphology of the axillary skin microbiota. Peptide nucleic acid - fluorescence in situ hybridization probes were used in combination with confocal laser scanning microscopy. Significant differences were found in both distribution and quantity of the cutaneous microbiota in clinically non-affected axillary skin of patients with HS compared with healthy controls. Surprisingly, we detected fewer bacteria and less biofilm in patients with HS. The reduced microbiota in patients with HS may play an important role in the early course of the disease.

Atomic Force Microscopy: A three-dimensional reconstructive tool of oral microbiota in gingivitis and periodontitis.

Aim:This study aims to ascertain the advantages of Atomic Force Microscopy (AFM) in the morphologic study of microorganisms and their interactions within the subgingival biofilm in patients with gingivitis and periodontitis.Settings and Design:Conducted a study on twenty patients, ten patients with severe periodontitis with probing the pocket depth of ≥8 mm, with a clinical attachment loss (CAL) of ≥6 mm CAL and ten patients with gingivitis: ≥5 mm pocket depth, and no attachment loss, was selected for the study.Materials and Methods:Bacterial biofilms were collected and slide preparation done. Morphological study was done using AFM. AFM consists of a cantilever-mounted tip, a piezoelectric scanner, a photodetector diode, a laser diode, and a feedback control. The laser beam is reflected from back of the cantilever into the quadrant of the photodetector. AFM works on the principle of interaction between the tip and the sample which causes the cantilever to deflect, thereby changing the position of laser onto the photodetector. Methodology used for studying the bacteria through AFM includes the following: (1) Probe type: Platinum coated silicon nitrate tip. (2) Probe force: 0.11 N/m. (3) Probe geometry: Triangular shaped tip. (4) Probe frequency: 22 KHz. (5) Probe immobilization: Used in Contact mode. AFM Solver Pro-M (NT-MDT) equipped with ETALON probe was used to take images in Nova software.Results:The investigation showed various morphological features, such as shape, size, and secretory product-like vesicles of the bacterial species involved in gingivitis and periodontitis. More bacterial surface details were studied by reproducing a three-dimensional reconstruction using AFM.Conclusions:The morphological variations of bacteria of different sizes, and shapes, cell wall structures, secretory product-like vesicles flagellated and filamentous microorganisms, polymorphonuclear leukocytes, and bacterial coaggregation analysis were done by AFM. Results of the present study conclude that AFM is a quite a reliable method for studying morphology of bacterial species involving periodontal diseases and is also used to study microbial interactions in biofilm.

BIOFILM BACTERIA PLAYS A ROLE IN CSOM PATHOGENESIS AND HAS SIGNIFICANT CORRELATION WITH UNSAFE TYPE CSOM

Chronic suppurative otitis media (CSOM) may cause severe morbidity and mortality and remains a major health problem worldwide. The incidence of CSOM in Indonesia (1994-1996) is estimated at about 8.36 million people and CSOM general prevalence is 3.8% (Helmi 2005). This study aims to prove the existence of bacterial biofilm in patients with safe type and hazard type CSOM from mastoidectomy. The design was observational analytic with cross sectional approach. This study was conducted at the Teratai Wards, IBP Dr. Soetomo Hospital, and Electron Microscopy Unit, Faculty of Medicine, Airlangga University Surabaya, from November 2013 to June 2014. Samples of pathological tissues were taken by consecutive sampling and bacterial biofilms examination was done by SEM. SEM results categorized the biofilm bacteria as positive or negative. Biofilm bacteria tested positive when it shows three-dimensional representation of bacteria with clear shapes and sizes and clusters, the formation of amorphous material consisting of glycocalyx material surrounding the bacteria, and surface attachment. Biofilm bacteria was regarded as negative when there is no bacterial cluster, glycocalyx and surface attachment. This study was performed on 33 CSOM patients. Samples were divided into unsafe type CSOM (17 patients) and safe type CSOM (16 patients). Positive biofilm bacteria was found in 12 patients with unsafe type (75%) and 6 patients with safe type (35.35%). Negative biofilm bacteria was found in 4 patients with the unsafe type (25%) and 11 patients with safe type (67.9%). Logistic regression analysis revealed p value = 0.027, indicating the biofilm bacteria have a significant correlation with unsafe type CSOM (p &lt;0.05). In conclusion, biofilm bacteria plays a role in CSOM pathogenesis of biofilm bacteria has significant correlation with unsafe type CSOM.

Biofilm Producing Staphylococcus epidermidis Strains Isolated From Clinical Samples in Tehran, Iran

Background: Staphylococcus epidermidis is well documented as an opportunistic pathogen causing biofilm in patients and healthy individuals. Objectives: The aim of this experimental study was to describe the antibiotic resistance patterns of biofilm producing S. epidermidis strains isolated from clinical samples in Tehran, Iran. Moreover, the role of different genes in biofilm formation was also described. Patients and Methods: A total of 250 S. epidermidis strains were isolated from patients in a private hospital of Tehran, Iran from February to December 2014. The biofilm formation of each strain was determined using combination of qualitative Congo-Red agar and quantitative microtiter plate assay, and presence of different genes involved in control and formation of biofilm was detected by the polymerase chain reaction (PCR). Susceptibility of S. epidermidis strains to 19 antibiotics was examined. Results: The results of the biofilm assay revealed that 82% of strains produced black colonies on Congo red agar plates and 68% were able to attach strongly to polystyrene microplates. One hundred percent, 88%, 84%, 64% and 60% of biofilm-producing strains were resistant to penicillin, cefoxitin, erythromycin, trimethoprim-sulfamethoxazole and kanamycin, respectively. On the other hand, none of the strains showed resistance to vancomycin, linezolid, quinupristin/dalfopristin. The icaA, icaD, aap and atlE genes were detected in all biofilm-producing strains and presence of IS256 transposon was limited to 84% of biofilm positive strains. Conclusions: The results of this study illustrated the high prevalence of antibiotic resistant biofilm-producing S. epidermidis strains in this hospital, which could be a reservoir for antibiotic resistance genes.

Prevalence of fungal biofilms in patients with nasal polyposis: a systematic review

Introduction: There are many internal and external factors that are considered as the main causes of nasal infections and inflammation in chronic rhinosinusitis (CRS) leading to polyposis. It is suggested that bacterial and fungal elements have an important role in the development of these diseases. Method: Scopus and PubMed were searched thoroughly on 1 February 2015 with the following search terms: (fungal biofilms) AND (nasal polyposis) to find the articles in which the prevalence of fungal biofilms had been evaluated in patients with nasal polyposis. Only English language articles with no time limitation were included in this study.Result: Of 48 records found by initial search, only 10 articles met the inclusion criteria. Data showed that the presence of biofilms is associated with nasal polyposis in 110 of 303 patients and 13 of controls.Discussion: The results of studies confirm that the fungal biofilms play an important role in the pathogenesis of chronic rhinosinusitis with nasal polyposis.Conclusions: According to the results of included studies, there is a close association between the presence of fungal biofilm and different types of nasal disorders including nasal polyposis.

Impact of osteitis and biofilm formation and correlation between both in diffuse sinonasal polyposis in Egyptian adults; a prospective clinical and histopathologic study

BackgroundThe pathogenesis of diffuse sinonasal polyposis is still not completely established, possible explanations are osteitis, aeroallergens, fungal sinusitis and biofilms. There are no reports in Egypt about osteitis and biofilms in those patients. PurposeTo study the incidence and impact of osteitis and biofilms in Egyptian patients on diffuse sinonasal polyposis patients. Patients and methodsFifty patients (22 males, mean age of 30.68±7.24years) submitted to surgery for diffuse sinonasal polyposis. Computerized scan on sinuses ordered and scored by Lund–Mackay staging protocol, severity of Osteitis using the Global Osteitis Scoring Scale. Tissue samples were taken from diseased sinuses to be analyzed histopathologically for osteitis, and with scanning electron microscopy to detect bacterial biofilms. Another ten patients as a control scheduled for septoplasty or turbinectomy with no evidence of sinusitis, and tissue specimens were obtained 1cm behind the anterior end of inferior turbinate and processed in the same manner for biofilm comparison. Study designContemporary prospective cross-sectional cohort study. ResultsIn 70% (35/50) of the polyposis patients, histopathology was positive for osteitis. Biofilms were detected by electron microscope in 39 (78%). Two of controls (20%) were biofilm positive with a significant difference (p=0.035). The mean Lund–Mackay was 19.08±3.67 and mean osteitis score was 18.68±11.99. There was a significant correlation between Lund–Mackay and osteitis score (p<0.001) and between both and histopathologically proven osteitis (p=0.049), biofilms (p=0.005) and postoperative endoscopic healing (p=0.046) where increased soft tissue disease and osteitis and biofilm were associated with bad healing and vice versa. ConclusionOsteitis and bacterial biofilms underlie the majority of Polypoidal chronic rhinosinusitis and both correlated significantly. Scanning electron microscope is a good tool for detecting bacterial biofilms. Sinus surgery with surgical ventilation, mechanical disruption of biofilms and osteitis is a mandatory therapeutic choice with prolonged treatment with antibiotics and nasal wash.

Bacterial biofilms in chronic rhinosinusitis; distribution and prevalence

Conclusion: Biofilms were more prevalent in patients with CRSwNP compared to both CRSsNP and controls, and also on the ethmoid bulla compared to the middle turbinate, supporting a biofilm-related pathogenesis of CRSwNP.Objective: To investigate the prevalence of biofilms in patients with chronic rhinosinusitis with nasal polyps (CRSwNP) compared to patients with chronic rhinosinusitis without nasal polyps (CRSsNP) and controls. To examine the prevalence of biofilms in different anatomical localizations.Study design: Cross-sectional.Methods: This study comprised 27 patients with CRSsNP, 34 patients with CRSwNP, and 25 controls. Biopsies from the middle turbinate, the uncinate process, and the ethmoid bulla were harvested pre-operatively, snap frozen in isopentane, cooled, and stored at −80°C. Prepared with Invitrogens’ Baclight LiveDead kit and investigated with confocal scanning laser microscopy.Results: Biofilms were studied in 33/34 (97%) CRSwNP, 22/27 (82%) CRSsNP, and 14/25 (56%) controls. The difference in point prevalence between patients with CRSwNP vs CRSsNP (p = 0.042, χ2 = 4.12), CRSwNP vs Controls (p < 0.001, χ2 = 15.0), and CRSsNP vs controls (p = 0.047, χ2 = 3.96) were all significant. Biofilms were found in 43/54 (80%) ethmoid bulla, 39/55 (71%) uncinate process, and 31/50 (62%) middle turbinate. The difference between the ethmoid bulla and the middle turbinate locations (p = 0.047, χ2 = 3.93) was significant.

The pathophysiological role of bacterial biofilms in chronic sinusitis.

Chronic rhinosinusitis (CRS) is a very common disorder that remains poorly understood from a pathogenic standpoint. Recent research on the pathogenesis of CRS has been focused on the potential role of biofilms in this chronic infection. The aim of this study was to assess the sinuses’ microflora and biofilm formation on the sino-nasal mucosa in patients with CRS. Paranasal sinus mucosa specimens were harvested at the time of functional endoscopic sinus surgery (FESS). Classical microbiology techniques for the isolation and identification of sinus mucosa microbial flora were used. Scanning electron microscopy (SEM) was used to detect biofilm on the surface of mucosa. A microtiter plate assay for in vitro biofilm formation was employed, divided into three aliquots. One part was assessed for bacterial presence, utilizing an API manual system and the Vitek® 2 Compact system. The two remaining aliquots were tested by in vitro conventional microbiological assay with the use of the Infinite M200 (Tecan) microtiter plate reader, and also by scanning electron microscopy (SEM). A microbiological examination of mucosal specimens had taken during FESS operation revealed the presence of various types of bacteria in 29 out of 30 tested samples. Out of 62 different strains isolated from patients with CRS, 23 strains of coagulase-negative Staphylococcus epidermidis and 6 strains of Escherichia coli were the most frequently isolated microorganisms, accounting for 37.1 and 9.7 %, respectively. Among the 62 isolated strains, 58 were used to assess biofilm formation. From the total of 58 isolates, 8.6 % were strong biofilm producers, 20.7 % were moderate, and 70.7 % of isolates were considered to be non- or weak biofilm producers. SEM of the 30 nasal concha mucosal samples taken from patients with CRS revealed biofilm in 23 specimens. A marked destruction of the epithelium was observed, with variation in degrees of severity, from disarrayed cilia to complete absence of cilia. The vast majority of nasal concha mucosal samples of patients affected by chronic sinusitis presented with biofilm formation. Our study showed that 76.7 % of patients having FESS for CRS had evidence of biofilms on SEM micrographs. Although certain detection methods could lead to various discrepancies in the amount of biofilm produced, the consistent demonstration of biofilms in patients with CRS suggests that this convoluted three-dimensional structures might play a significant role in either the pathogenesis or persistence of chronic rhinosinusitis.

Sinus biofilms in patients with cystic fibrosis: is adjusted eradication therapy needed?

The paranasal sinuses can be a focus for colonisation of the cystic fibrosis (CF) lungs with pathogens. In the sinuses, bacteria can adapt to the lung environment and enhance their antibiotic resistance, with biofilm formation thought to be the most important adaptive mechanism, causing recalcitrant disease. The presence of biofilms in CF sinuses is sparsely described. In this descriptive cross-sectional study, the sinus mucosa from 16 CF patients were analysed by fluorescence in situ hybridization using specific peptide nucleic acid (PNA-FISH) probes for Pseudomonas aeruginosa and Staphylococcus aureus to demonstrate the presence of biofilms. Small clusters of biofilm were visualised lining the sinus mucosa of CF patients. Biofilms were found in 10 out of 18 cases; 7 with intermittent lung colonisation, 2 chronically infected, and one lung transplanted patient. Finding P. aeruginosa biofilms in intermittently lung-colonised patients encourage us to intensify the attempt to eradicate pathogenic bacteria from the CF sinuses in an early stage using combined antibiotic therapy in the prolonged exposure of the sinus-mucosal surface.

Biofilm Formation and its Impact on Antifungal Therapy

Fungi can protect themselves from host defences and antifungal drugs by the production of an extracellular hydrophobic matrix. Candida biofilms exhibit resistance to antifungal agents from all classes including the azoles, echinocandins, amphotericin B complex, and flucytosine. Although demonstrated on polystyrene and bronchial epithelia cells, until today, only indirect evidence for A. fumigatus biofilms in patients is available. The antifungals with the most activity against biofilms are the liposomal formulation of amphotericin B and agents in the echinocandin drug class. Importantly, echinocandins show excellent anti-biofilm activity against C. albicans at therapeutic concentrations. However, other biofilms formed by moulds, including A. fumigatus, are relatively resistant to echinocandins. Multiple mechanisms contribute to the intrinsic and acquired antifungal resistance during the different stages of fungal biofilm development. During the growth phase of the early biofilm various factors account for biofilm resistance. Combinational and sequential antifungal therapy as well as combination with enhancers can improve the effect of a single drug. Further studies are warranted to develop new therapeutic strategies targeting fungal biofilm-specific resistance mechanisms.

Bacterial biofilms in patients with chronic rhinosinusitis: a confocal scanning laser microscopy study.

Recent research into the pathophysiology of chronic rhinosinusitis suggests an important role for biofilms. They can be detected in both healthy and diseased nasal mucosa. Several different methods of detecting biofilms have been described. This study investigates the presence of biofilm in a larger group of patients with chronic rhinosinusitis undergoing primary functional endoscopic surgery. Sixty-one patients with chronic rhinosinusitis and 25 controls, with septal deviation, were included from 2010 to 2012. Endonasal biopsies were harvested during surgery, snap frozen in isopentane, cooled on dry ice and stored at -80oC. The samples were prepared with Invitrogens’ BacLight LiveDead kit, and investigated with confocal scanning laser microscopy for the presence of biofilm. In the chronic rhinosinusitis group 55/61 were biofilm positive as opposed to 14/25 in the control group. The difference was highly significant. The odds ratio was 7.2. Patients with chronic rhinosinusitis have a highly significant increased point prevalence of biofilms compared to controls.

G385 RATNO – Reducing Antibiotic Tolerance using Nitric Oxide in Cystic Fibrosis: report of a proof of concept clinical trial

AimsTo investigate whether low dose Nitric Oxide (NO) enhances antibiotic therapy through disruption of Pseudomonas aeruginosa (PA) bacteria in biofilms in patients with Cystic Fibrosis (CF).BackgroundBacterial biofilms present a major...

Clinical characteristics of biofilms in patients with chronic rhinosinusitis: a prospective case-control study.

Recent studies have indicated that biofilms are involved in the pathogenesis of recurrent and recalcitrant chronic rhinosinusitis (CRS). The present study was performed to evaluate the presence of biofilms and to evaluate the relationships between the presence of biofilms and clinical features of CRS. A total of 33 patients were included in this study. Maxillary sinus mucosa from 26 CRS patients and the ethmoid mucosa from 7 patients undergoing septoplasty were collected. Biofilms were evaluated by scanning electron microscopy. Preoperative symptom scores, preoperative and intraoperative nasoendoscopic findings, and postoperative healing period were compared between the groups. Biofilms were detected in 14 (42.4%) of the 33 patients. Biofilms were present in 13 (50%) of the 26 patients in the CRS group, but in only one (14.3%) of the seven patients in the control group. There were no significant differences in preoperative symptom scores or preoperative nasal Lund-Kennedy endoscopic scores between the groups. However, the average Lund-Mackay and intraoperative sinus Lund-Kennedy endoscopic scores were significantly higher in biofilm-positive than biofilm-negative patients (P<0.05). In the follow-up period, the healing time was significantly longer in biofilm-positive than biofilm-negative patients (P<0.05). This study suggested that the presence of bacterial biofilms may contribute to the pathogenesis of CRS and the clinical characteristics of CRS patients after endoscopic sinus surgery.