John G. Thomas

Chronic rhinosinusitis and biofilms.

BACKGROUND AND HYPOTHESIS Biofilms have been implicated in several head and neck infectious processes such as the following: dental and periodontal disease, otitis media, tympanostomy tube otorrhea, and chronic tonsillitis. We believe that biofilms also are associated with chronic rhinosinusitis. No information is known regarding the presence of biofilms in chronic rhinosinusitis. STUDY DESIGN AND SETTING With institutional review board approval, tissue was obtained from consenting chronic rhinosinusitis patients who were undergoing functional endoscopic sinus surgery. Specimens were taken bilaterally from the ethmoid and maxillary sinuses. Inclusion criteria consisted of a positive diagnosis with pathologic tissue confirmation of chronic inflammation. Diagnosis was based on patient history, physical exam, and coronal sinus CT findings. Once collected, the specimens were labeled and fixed in formalin. The specimens were subsequently dehydrated, with successive immersions in increasing concentrations of diluted ethanol. The specimens were allowed to air dry and then were affixed to aluminum stubs with colloidal carbon. The sample surface was coated with a gold and palladium layer. The specimens were examined under an electron microscope. Areas of interest were photographed. RESULTS Specimens from 5 patients were examined. All revealed bacterial biofilms. Invariably, biofilms were seen in the ethmoid, as well as in other samples. Denudation of ciliated and goblet cells was noted in all specimens. Biofilms resembled that of Staphylococcus species. Unidentified biofilms were also seen. CONCLUSIONS This is the first documentation of biofilms in association with chronic rhinosinusitis. Further investigation is warranted, especially with control research subjects.

Biofilms and bacterial imbalances in chronic wounds: anti-Koch.

Microbial imbalances and synergistic relationships between bacteria in medically important biofilms are poorly researched. Consequently, little is known about how synergy between bacteria may increase the net pathogenic effect of a biofilm in many diseases and infections, including chronic wounds. Microbial synergy in chronic wounds may increase virulence and pathogenicity, leading to enhanced tissue degradation, malodour and in some cases, an impairment of the host immune response. Microbial synergy and growth within a biofilm provide a competitive advantage to the microorganisms cohabiting in a wound, thereby promoting their survival and tolerance and resistance to antimicrobial agents. The aim of this article was to provide greater insight into microbial imbalances found within wound biofilms and the significance they may have on non healing and infected wounds. We also present two possible hypotheses which could explain the role microorganisms play in non healing chronic wounds and offer possible strategies for combating harmful and detrimental biofilms.

Study protocol of the Center for Oral Health Research in Appalachia (COHRA) etiology study

BackgroundPeople in Appalachia experience some of the worst oral health in the United States. To develop effective intervention and prevention strategies in Appalachia, we must understand the complex relationships among the contributing factors and how they affect the etiology of oral diseases. To date, no such comprehensive analysis has been conducted. This report summarizes the characteristics of the sample and describes the protocol of a study determining contributions of individual, family, and community factors to oral diseases in Appalachian children and their relatives.Methods/DesignFamilies participated in a comprehensive assessment protocol involving interviews, questionnaires, a clinical oral health assessment, a microbiological assessment, and collection of DNA. The design of the study is cross-sectional.ConclusionDue to its multilevel design and large, family-based sample, this study has the potential to greatly advance our understanding of factors that contribute to oral health in Appalachian children.

The antimicrobial efficacy of a silver alginate dressing against a broad spectrum of clinically relevant wound isolates

Wound dressings impregnated with silver have a role to play in aiding to reduce both the dressing and wound microbial bioburden. It is therefore imperative that antimicrobial wound dressings have efficacy on a broad range of clinical significant microorganisms. Accordingly, this study aimed to determine the antimicrobial efficacy of a silver alginate dressing against 115 wound isolates that had been isolated routinely from patients at West Virginia University Hospital. Standardised corrected zones of inhibition (CZOIs) were performed on all clinical isolates. It was found that the silver alginate dressing was able to inhibit the growth of all microorganisms tested. In particular, the silver alginate dressing inhibited the growth of Candida albicans and yeasts with CZOI of 3–11·5 mm. All meticillin‐resistant Staphylococcus aureus (MRSA) strains were found to be sensitive to the silver alginate dressing with a CZOI range calculated at 3–7·8 mm. Sensitivity to the silver alginate dressing was also evident for S. aureus and vancomycin‐resistant Enterococci. CZOIs of 4·25 mm were calculated for Enterococcus faecium and 9·8 mm for viridans streptococcus. The bacteria which demonstrated the highest tolerance to ionic silver included Enterobacter cloacae and Acinetobacter baumannii. Contrary to this the most responsive microorganisms to ionic silver included strains of staphylococci, viridans streptococcus and Candida albicans. No antibiotic‐resistant isolates, as identified by Kirby Bauer Clinical Laboratory Standards Institute classification system, were found to be resistant to ionic silver. When a selected number of microorganisms were grown in the biofilm phenotypic state enhanced tolerance to silver was observed, compared to their non biofilm counterparts. Overall, this study has demonstrated the broad antimicrobial activity of a silver alginate dressing on wound isolates grown in the non biofilm and biofilm state. This finding is clinically relevant as both the non biofilm and biofilm phenotypic states of microorganisms are evident in wounds and therefore significant to delayed healing. Consequently, it is imperative that antimicrobial wound dressings demonstrate antimicrobial activity against microorganisms in both phenotypic states.

Transmission of Helicobacter pylori and the role of water and biofilms.

Documented evidence relating to the survival of Helicobacter pylori outside the gastric niche is extremely limited. To date the primary transmission routes of H. pylori have yet to be confirmed and when this is achieved preventive infection control measures can be implemented to reduce and ultimately prevent human infection from this pathogen. There is mounting evidence which suggests that the prevalence of H. pylori infection has a strong correlation with access to clean water, suggesting a transmission route to the host. However, there are no established culture methods for the detection of viable H. pylori in the environment, in particular drinking water supplies, preventing the development of true epidemiological and risk assessments. The aim of this review is to highlight the available data to date that suggests drinking water and possible survival in biofilms as a probable transmission mode for H. pylori.

Increases in Endotracheal Tube Resistance Are Unpredictable Relative to Duration of Intubation

BACKGROUND Accumulated secretions after intubation can affect the resistance of an endotracheal tube (ETT). Our objective was to measure extubated patient tubes and size-matched controls to evaluate differences in resistance. METHODS New ETTs, with internal diameters of 7.0 through 8.5 mm, were tested as controls to establish the resistance of each size group as measured by pressure drop. Measurements were obtained using a mass flowmeter and pressure transducer. Pressure drop was measured at three flow rates. Seventy-one patient ETTs were evaluated after extubation by an identical method and compared with controls. RESULTS In each control group, pressure drop was tightly clustered with low variation and no overlap between sizes. A total of 73 to 79% of the patient ETTs had a pressure drop of > 3 SDs of size-matched controls at all flow rates. Pressure drop in 48 to 56% (across three flow rates) of extubated tubes was equivalent to the next smaller size of controls. At 60 and 90 L/min, 10% and 15% of patient tubes, respectively, had the pressure drop of a control tube three sizes smaller. The pressure drop was unpredictable relative to the duration of intubation. CONCLUSIONS Organized secretions can significantly increase resistance as measured by the pressure drop of ETTs. The degree of change was highly variable, occurs in all sizes, and was unrelated to the duration of intubation. The performance of an ETT may be comparable to new tubes one to four sizes smaller. This may impact the tolerance of ventilator weaning.

Molecular analysis of microbial communities in endotracheal tube biofilms

Background Ventilator-associated pneumonia is the most prevalent acquired infection of patients on intensive care units and is associated with considerable morbidity and mortality. Evidence suggests that an improved understanding of the composition of the biofilm communities that form on endotracheal tubes may result in the development of improved preventative strategies for ventilator-associated pneumonia. Methodology/Principal Findings The aim of this study was to characterise microbial biofilms on the inner luminal surface of extubated endotracheal tubes from ICU patients using PCR and molecular profiling. Twenty-four endotracheal tubes were obtained from twenty mechanically ventilated patients. Denaturing gradient gel electrophoresis (DGGE) profiling of 16S rRNA gene amplicons was used to assess the diversity of the bacterial population, together with species specific PCR of key marker oral microorganisms and a quantitative assessment of culturable aerobic bacteria. Analysis of culturable aerobic bacteria revealed a range of colonisation from no growth to 2.1×108 colony forming units (cfu)/cm2 of endotracheal tube (mean 1.4×107 cfu/cm2). PCR targeting of specific bacterial species detected the oral bacteria Streptococcus mutans (n = 5) and Porphyromonas gingivalis (n = 5). DGGE profiling of the endotracheal biofilms revealed complex banding patterns containing between 3 and 22 (mean 6) bands per tube, thus demonstrating the marked complexity of the constituent biofilms. Significant inter-patient diversity was evident. The number of DGGE bands detected was not related to total viable microbial counts or the duration of intubation. Conclusions/Significance Molecular profiling using DGGE demonstrated considerable biofilm compositional complexity and inter-patient diversity and provides a rapid method for the further study of biofilm composition in longitudinal and interventional studies. The presence of oral microorganisms in endotracheal tube biofilms suggests that these may be important in biofilm development and may provide a therapeutic target for the prevention of ventilator-associated pneumonia.

Long-Term Sub-Antimicrobial Doxycycline (Periostat®) as Adjunctive Management in Adult Periodontitis: Effects on Subgingival Bacterial Population Dynamics

Previous trials had indicated that various schedules of sub-antimicrobial doxycycline significantly reduced gingival crevicular fluid (GCF) collagenase activity in adult patients with periodontitis with no evidence of emergent tetracycline-resistant (Tcr) marker oral flora. The purpose of this nine-month study was to expand these observations, emphasizing newer microbial diagnostic methods. Subgingival paper point samples were obtained at baseline (BL), 3, 6, and 9 months. Four subject treatment groups in a double-blind design were evaluated by mechanical scaling and root planing (SRP) and/or 20 mg doxycycline BID (Periostat®). Thirty-eight patients entered the study at baseline (BL). Dark-field microscopy on 260 samples showed that morphotype distribution was independent of treatment schedule. Culture analysis of the 3 most prevalent isolates recovered showed that Streptococcus and Prevotella species accounted for approximately 85% of the 724 cultures. There did not appear to be any overgrowth or replacement by opportunistic oral flora. Of 658 susceptibility patterns evaluated by Etest, the MIC 50/90 and mode MIC showed stable patterns, independent of treatment group. Our findings were different from those of previously published reports, but may be partly explained by the lack of universally standardized methods in oral microbiology and interpretive criteria for susceptibility testing.

The antimicrobial efficacy of silver on antibiotic‐resistant bacteria isolated from burn wounds

The antibiotic‐resistant bacteria are a major concern to wound care because of their ability to resist many of the antibiotics used today to treat infections. Consequently, other antimicrobials, in particular ionic silver, are considered ideal topical agents for effectively helping to manage and prevent local infections. Little is known about the antimicrobial efficacy of ionic silver on antibiotic‐resistant bacteria at different pH values. Consequently, in this study our aim was to evaluate the effect of pH on the antimicrobial efficacy of a silver alginate (SA) and a silver carboxymethyl cellulose (SCMC) dressing on antibiotic‐resistant bacteria isolated from burn patients. Forty‐nine antibiotic‐resistant bacteria, including Vancomycin‐resistant Enterococcus faecium, meticillin‐resistant Staphylococcus aureus, multidrug‐resistant (MDR) Pseudomonas aeruginosa, MDR Vibrio sp, MDR Stenotrophomonas maltophilia, extended‐spectrum ß‐lactamase (ESBL) producing Salmonella sp, ESBL producing Klebsiella pneumoniae, ESBL producing Proteus mirabilis, ESBL producing Escherichia coli and MDR Acinetobacter baumannii, routinely isolated from burn wounds were used in the study and evaluated for their susceptibility to two silver containing wound dressings using a standardised antimicrobial efficacy screening assay [corrected zone of inhibition (CZOI)]. The mean overall CZOI for the Gram‐positive isolates at a pH of 5·5 were very similar for both dressings. A mean CZOI of 5 mm was recorded for the SCMC dressing, which was slightly higher, at 5·4 mm for the SA dressing. At a pH of 7·0 both dressings, in general, showed a similar activity. However, at a pH of 8·5 the mean CZOI of the SCMC dressing was found to be significantly (P < 0·05) higher than the SA dressing for a select number of isolates. The mean overall CZOI for the Gram‐negative bacteria followed a similar pattern as observed with the Gram‐positive bacteria. Susceptibility to silver ions did vary significantly between genera and species of bacteria. Interestingly, when pH was changed from 8·5 to 5·5 antimicrobial activity for both dressings in general increased significantly (P < 0·05). Overall, all forty‐nine antibiotic‐resistant bacteria isolated from burn wounds showed susceptibility to the antimicrobial activity of both silver containing wound dressings over all pH ranges. In addition, the study showed that the performance of both dressings apparently increased when pH became more acidic. The findings in this study may help to further enhance our knowledge of the role pH plays in affecting both bacterial susceptibility and antimicrobial activity of silver containing wound dressings.

Impact of a Multimodal Antimicrobial Stewardship Program on Pseudomonas aeruginosa Susceptibility and Antimicrobial Use in the Intensive Care Unit Setting

Objective. To study the impact of our multimodal antibiotic stewardship program on Pseudomonas aeruginosa susceptibility and antibiotic use in the intensive care unit (ICU) setting. Methods. Our stewardship program employed the key tenants of published antimicrobial stewardship guidelines. These included prospective audits with intervention and feedback, formulary restriction with preauthorization, educational conferences, guidelines for use, antimicrobial cycling, and de-escalation of therapy. ICU antibiotic use was measured and expressed as defined daily doses (DDD) per 1,000 patient-days. Results. Certain temporal relationships between antibiotic use and ICU resistance patterns appeared to be affected by our antibiotic stewardship program. In particular, the ICU use of intravenous ciprofloxacin and ceftazidime declined from 148 and 62.5 DDD/1,000 patient-days to 40.0 and 24.5, respectively, during 2004 to 2007. An increase in the use of these agents and resistance to these agents was witnessed during 2008–2010. Despite variability in antibiotic usage from the stewardship efforts, we were overall unable to show statistical relationships with P. aeruginosa resistance rate. Conclusion. Antibiotic resistance in the ICU setting is complex. Multimodal stewardship efforts attempt to prevent resistance, but such programs clearly have their limits.