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PubMed2001

Characteristics of biofilm formation by Candida albicans.

A variety of manifestations of Candida albicans infections are associated with the formation of biofilms on the surface of biomaterials. Cells in biofilms display phenotypic traits that are dramatically different from their free-floating planktonic counterparts, such as increased resistance to anti-microbial agents and protection form host defenses. Here, we describe the characteristics of C. albicans biofilm development using a 96 well microtitre plate model, microscopic observations and a colorimetric method based on the use of a modified tetrazolium salt (2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide, XTT) to monitor metabolic activities of cells within the biofilm. C. albicans biofilm formation was characterized by initial adherence of yeast cells (0-2 h), followed by germination and micro-colony formation (2-4 h), filamentation (4-6 h), monolayer development (6-8 h), proliferation (8-24 h) and maturation (24-48 h). The XTT-reduction assay showed a linear relationship between cellular density of the biofilm and metabolic activity. Serum and saliva pre-conditioning films increased the initial attachment of C. albicans, but had minimal effect on subsequent biofilm formation. Scanning electron microscopy and confocal scanning laser microscopy were used to visualize C. albicans biofilms. Mature C. albicans biofilms consisted of a dense network of yeasts cells and hyphal elements embedded within exopolymeric material. C. albicans biofilms displayed a complex three dimensional structure which demonstrated spatial heterogeneity and a typical architecture showing microcolonies with ramifying water channels. Antifungal susceptibility testing demonstrated the increased resistance of sessile C. albicans cells against clinically used fluconazole and amphotericin B as compared to their planktonic counterparts.

Journal Article↗
PubMed2003

Anatomical evidence of microbial biofilms in tonsillar tissues: a possible mechanism to explain chronicity.

CONTEXT: Bacteria within biofilms are resistant to host defenses and antibiotics. The presence of bacterial biofilms within the tissue and crypts of inflamed tonsils may explain the chronicity and recurrent characteristics of some forms of tonsillitis. OBJECTIVE: To determine if microbial biofilms occur within clinically abnormal tonsils. DESIGN: In this study, we evaluated the histomorphological appearance of 19 human tonsils for evidence of biofilm formation using light and transmission electron microscopy. SUBJECTS: Human tissues were collected during surgical tonsillectomy. Fifteen specimens were removed because of a history of repeated infections, and 4 were removed because of hypertrophy and obstruction. INTERVENTIONS: No interventions were used in this study. MAIN OUTCOME MEASURE: Histological and ultrastructural evidence of bacterial biofilms within the crypts of tonsils. RESULTS: Gram-positive and gram-negative bacteria were seen within otherwise acellular deposits among crypts of 11 of 15 infected tonsils. Regions of accumulated bacteria possessed the ultrastructural appearance of typical amorphous polysaccharide biofilm matrix. Small clusters of bacterial colonies were seen in 3 of 4 tonsils removed because of hypertrophy. CONCLUSIONS: There is strong anatomical evidence for the presence of bacterial biofilms in chronically diseased tonsils. Because sessile bacteria within biofilms are resistant to host defenses and antibiotics, bacterial biofilms within tonsils may explain the chronicity and recurrent nature of some forms of tonsillitis.

Biofilms↗
PubMed2006

Hha, YbaJ, and OmpA regulate Escherichia coli K12 biofilm formation and conjugation plasmids abolish motility.

Escherichia coli Hha is an environmental-response regulator of the pathogenic hemolysin operon, and Hha and the contiguous YbaJ are both induced 30-fold in E. coli biofilms (Appl. Microbiol. Biotechnol. 64:515, 2004). Here it is shown that Hha and YbaJ regulate biofilm formation since the hha/ybaJ deletion reduces biofilm mass in microtitre plates (81% in minimal medium, 50% in complex medium) and in flow cells (1,000-fold less surface coverage in minimal medium). The addition of the derepressed conjugative plasmid R1drd19, which increases significantly biofilm formation, eliminated motility completely in wild-type E. coli K12, promoted cell aggregation 27.18 +/- 0.05-fold, and produced a flatter biofilm. Deletion of hha/ybaJ or ybaJ restored motility (this motility phenotype may be complemented by providing hha(+)/ybaJ(+) or ybaJ(+) in trans) and reduced cell aggregation to that of the wild-type strain that lacks the conjugation plasmid. This increase in motility due to deleting hha/ybaJ was found to be due to 8-fold induction of fliA transcription. In addition, deletion of ompA reduced biofilm mass by 80% in both LB medium and LB medium with glucose. Also, Hha/YbaJ promotes conjugation since there was five-fold less conjugation in the hha/ybaJ mutant. It appears that conjugation plasmids promote biofilm formation by promoting cell aggregation, and that Hha and YbaJ increase biofilm formation by increasing conjugation and by decreasing motility when a conjugative plasmid (R1drd19) is present (YbaJ plays the most important role in this regulation of motility). When hha/ybaJ are deleted, there is less conjugation, less aggregation, more motility, and less biofilm.

Biofilms↗
PubMed2006

Three-dimensional biofilm model with individual cells and continuum EPS matrix.

An innovative type of biofilm model is derived by combining an individual description of microbial particles with a continuum representation of the biofilm matrix. This hybrid model retains the advantages of each approach, while providing a more realistic description of the temporal development of biofilm structure in two or three spatial dimensions. The general model derivation takes into account any possible number of soluble components. These are substrates and metabolic products, which diffuse and react in the biofilm within individual microbial cells. The cells grow, divide, and produce extracellular polymeric substances (EPS) in a multispecies model setting. The EPS matrix is described by a continuum representation as incompressible viscous fluid, which can expand and retract due to generation and consumption processes. The cells move due to a pushing mechanism between cells in colonies and by an advective mechanism supported by the EPS dynamics. Detachment of both cells and EPS follows a continuum approach, whereas cells attach in discrete events. Two case studies are presented for model illustration. Biofilm consolidation is explained by shrinking due to EPS and cell degradation processes. This mechanism describes formation of a denser layer of cells in the biofilm depth and occurrence of an irregularly shaped biofilm surface under nutrient limiting conditions. Micro-colony formation is investigated by growth of autotrophic microbial colonies in an EPS matrix produced by heterotrophic cells. Size and shape of colonies of ammonia and nitrite-oxidizing bacteria (NOB) are comparatively studied in a standard biofilm and in biofilms aerated from a membrane side.

Biofilms↗
PubMed2007

Biocidal effect of cathodic protection on bacterial viability in biofilm attached to carbon steel.

Biofilm formed on carbon steel by various species of bacterial cells causes serious problems such as corrosion of steel, choking of flow in the pipe, deterioration of the heat-transfer efficiency, and so on. Cathodic protection is known to be a reliable method for protecting carbon steel from corrosion. However, the initial attachment of bacteria to the surface and the effects of cathodic protection on bacterial viability in the biofilm have not been clarified. In this study, cathodic protection was applied to an artificial biofilm containing Pseudomonas aeruginosa (PAO1), a biofilm constituent, on carbon steel. The aims of this study were to evaluate the inhibition effect of cathodic protection on biofilm formation and to reveal the inhibition mechanisms. The viability of PAO1 in artificial biofilm of 5 mm thickness on cathodically protected steel decreased to 1% of the initial cell concentration. Analysis of pH distribution in the artificial biofilm by pH microelectrode revealed that pH in proximity to carbon steel increased to approximately 11 after cathodic protection for 5 h. Moreover, 99% of region in the artificial biofilm was under the pH conditions of over nine. A simulation of pH profile was shown to correspond to experimental values. These results indicate cells in the artificial biofilm were killed or damaged by cathodic protection due to pH increase.

Biofilms↗
PubMed2006

Biofilms.

Biofilms of bacteria, frequently in association with algae, protozoa and fungi, are found on all submerged structures in the marine environment. Although it is likely that for the majority of organisms a biofilmed surface is not a pre-requisite for settlement, in practice, colonization by spores and larvae of fouling organisms almost always takes place via a biofilmed surface. Therefore, the properties of the latter may be expected to influence colonization, positively or negatively. Biofilms are responsible for a range of surface-associated and diffusible signals, which may moderate the settling behaviour of cells, spores and larvae. However, there is no consensus view regarding either cause and effect or the mechanism(s) by which biofilms moderate settlement. Studies with mixed biofilms, especially field experiments, are difficult to interpret because of the conflicting signals produced by different members of the biofilm community as well as their spatial organisation. Molecular techniques highlight the deficiencies of culture methods in identifying biofilm bacteria; hence, the strains with the most impact on settlement of spores and larvae may not yet have been isolated and cultured. Furthermore, secondary products isolated from cultured organisms may not reflect the situation that pertains in nature. The evidence that bacterial quorum sensing signal molecules stimulate settlement of spores of the green macroalga, Ulva, is discussed in some detail. New molecular and analytical tools should provide the opportunity to improve our fundamental understanding of the interactions between fouling organisms and biofilms, which in turn may inform novel strategies to control biofouling.

4-Butyrolactone↗
PubMed2002

Microscale evaluation of the effects of grazing by invertebrates with contrasting feeding modes on river biofilm architecture and composition.

River biofilms are a valuable food resource for many invertebrates. In the present study biofilms were cultivated in a rotating annular bioreactor with river water as sole source of inoculum. The resulting biofilms were then presented to starved snails, ostracods, and mayflies as sole food source. The biofilms were then removed and microscopically examined to determine areas that had been grazed. The grazed and ungrazed areas were marked and analyzed for the effects of grazing using confocal laser scanning microscopy and image analyses. Samples were treated with fluorescent probes for nucleic acids to quantify bacterial biomass and fluor-conjugated lectins to quantify exopolymer, and far red autofluorescence was imaged to quantify algal or photosynthetic biomass. Grazing by snails significantly reduced algal biomass (1.1 +/- 0.6 micro m 3 micro m 2 to 0.02 +/- 0.04 micro m 3 micro m 2), exopolymer (5.3 +/- 3.4 micro m 3 micro m 2 to 0.18 +/- 0.18 micro m 3 micro m 2), and biofilm thickness (154 micro m +/- 50 to 11 micro m +/- 5.2; ANOVA, p < or= 0.05). Although bacterial biomass was influenced by grazing snails the impact was not statistically significant (p <or= 0.05). Ostracods had a significant (p <or= 0.05) impact on the algal biomass and exopolymer but not on bacteria. Mayfly grazing resulted in reduction of biofilm thickness to approximately 40 mm and reductions in all biofilm components with little evidence of selectivity. Thus grazing consistently resulted in a significant reduction in autotrophic biomass and exopolymer with a resultant increased importance of bacterial biomass within the grazed regions. Examination of grazed biofilms after a recovery period in the absence of grazing indicated that grazed regions remained predominantly bacterial after 28 days. A comparison of grazing in diclofop methyl and atrazine contaminated (1 ppb and 10 ppb) versus control biofilms indicated no significant influence of the contaminants on grazing patterns.

Animals↗
PubMed2005

Interaction of Klebsiella oxytoca and Burkholderia cepacia in dual-species batch cultures and biofilms as a function of growth rate and substrate concentration.

Dual-species microbial interactions have been extensively reported for batch and continuous culture environments. However, little research has been performed on dual-species interaction in a biofilm. This research examined the effects of growth rate and substrate concentration on dual-species population densities in batch and biofilm reactors. In addition, the feasibility of using batch reactor kinetics to describe dual-species biofilm interactions was explored. The scope of the research was directed toward creating a dual-species biofilm for the biodegradation of trichloroethylene, but the findings are a significant contribution to the study of dual-species interactions in general. The two bacterial species used were Burkholderia cepacia PR1-pTOM(31c), an aerobic organism capable of constitutively mineralizing trichloroethylene (TCE), and Klebsiella oxytoca, a highly mucoid, facultative anaerobic organism. The substrate concentrations used were different dilutions of a nutrient-rich medium resulting in dissolved organic carbon (DOC) concentrations on the order of 30, 70, and 700 mg/L. Presented herein are single- and dual-species population densities and growth rates for these two organisms grown in batch and continuous-flow biofilm reactors. In batch reactors, planktonic growth rates predicted dual-species planktonic species dominance, with the faster-growing organism (K. oxytoca) outcompeting the slower-growing organism (B. cepacia). In a dual-species biofilm, however, dual-species planktonic growth rates did not predict which organism would have the higher dual-species biofilm population density. The relative fraction of each organism in a dual-species biofilm did correlate with substrate concentration, with B. cepacia having a greater proportional density in the dual-species culture with K. oxytoca at low (30 and 70 mg/L DOC) substrate concentrations and K. oxytoca having a greater dual-species population density at a high (700 mg/L DOC) substrate concentration. Results from this research demonstrate the effectiveness of using substrate concentration to control population density in this dual-species biofilm.

Biodegradation, Environmental↗
PubMed2005

Molecular assessment of inoculated and indigenous bacteria in biofilms from a pilot-scale perchlorate-reducing bioreactor.

Bioremediation of perchlorate-contaminated groundwater can occur via bacterial reduction of perchlorate to chloride. Although perchlorate reduction has been demonstrated in bacterial pure cultures, little is known about the efficacy of using perchlorate-reducing bacteria as inoculants for bioremediation in the field. A pilot-scale, fixed-bed bioreactor containing plastic support medium was used to treat perchlorate-contaminated groundwater at a site in Southern California. The bioreactor was inoculated with a field-grown suspension of the perchlorate-respiring bacterium Dechlorosoma sp. strain KJ and fed groundwater containing indigenous bacteria and a carbon source amendment. Because the reactor was flushed weekly to remove accumulated biomass, only bacteria capable of growing in biofilms in the reactor were expected to survive. After 26 days of operation, perchlorate was not detected in bioreactor effluent. Perchlorate remained undetected by ion chromatography (detection limit 4 mug L(-1)) during 6 months of operation, after which the reactor was drained. Plastic medium was subsampled from top, middle, and bottom locations of the reactor for shipment on blue ice and storage at -80 degrees C prior to analysis. Microbial community DNA was extracted from successive washes of thawed biofilm material for PCR-based community profiling by 16S-23S ribosomal intergenic spacer analysis (RISA). No DNA sequences characteristic of strain KJ were recovered from any RISA bands. The most intense bands yielded DNA sequences with high similarities to Dechloromonas spp., a closely related but different genus of perchlorate-respiring bacteria. Additional sequences from RISA profiles indicated presence of representatives of the low G+C gram-positive bacteria and the Cytophaga-Flavobacterium-Bacteroides group. Confocal scanning laser microscopy and fluorescence in situ hybridization (FISH) were also used to examine biofilms using genus-specific 16S ribosomal RNA probes. FISH was more sensitive than RISA profiling in detecting possible survivors from the initial inoculum. FISH revealed that bacteria hybridizing to Dechlorosoma probes constituted <1% of all cells in the biofilms examined, except in the deepest portions where they represented 3-5%. Numbers of bacteria hybridizing to Dechloromonas probes decreased as biofilm depth increased, and they were most abundant at the biofilm surface (23% of all cells). These spatial distribution differences suggested persistence of low numbers of the inoculated strain Dechlorosoma sp. KJ in parts of the biofilm nearest to the plastic medium, concomitant with active colonization or growth by indigenous Dechloromonas spp. in the biofilm exterior. This study demonstrated the feasibility of post hoc analysis of frozen biofilms following completion of field remediation studies.

Bacteria↗
PubMed2003

Sulfate-reducing bacterial community structure and their contribution to carbon mineralization in a wastewater biofilm growing under microaerophilic conditions.

The community structure of sulfate-reducing bacteria (SRB) and the contribution of SRB to carbon mineralization in a wastewater biofilm growing under microaerophilic conditions were investigated by combining molecular techniques, molybdate inhibition batch experiments, and microelectrode measurements. A 16S rDNA clone library of bacteria populations was constructed from the biofilm sample. The 102 clones analyzed were grouped into 53 operational taxonomic units (OTUs), where the clone distribution was as follows: Cytophaga- Flexibacter- Bacteroides (41%), Proteobacteria (41%), low-G+C Gram-positive bacteria (18%), and other phyla (3%). Three additional bacterial clone libraries were also constructed from SRB enrichment cultures with propionate, acetate, and H(2) as electron donors to further investigate the differences in SRB community structure due to amendments of different carbon sources. These libraries revealed that SRB clones were phylogenetically diverse and affiliated with six major SRB genera in the delta-subclass of the Proteobacteria. Fluorescent in situ hybridization (FISH) analysis revealed that Desulfobulbus and Desulfonema were the most abundant SRB species in this biofilm, and this higher abundance (ca. 2-4x10(9) cells cm(-3) and 5x10(7) filaments cm(-3), respectively) was detected in the surface of the biofilm. Microelectrode measurements showed that a high sulfate-reducing activity was localized in a narrow zone located just below the oxic/anoxic interface when the biofilm was cultured in a synthetic medium with acetate as the sole carbon source. In contrast, a broad sulfate-reducing zone was found in the entire anoxic strata when the biofilm was cultured in the supernatant of the primary settling tank effluent. This is probably because organic carbon sources diffused into the biofilm from the bulk water and an unknown amount of volatile fatty acids was produced in the biofilm. A combined approach of molecular techniques and batch experiments with a specific inhibitor (molybdate) clearly demonstrated that Desulfobulbus is a numerically important member of SRB populations and the main contributor to the oxidation of propionate to acetate in this biofilm. However, acetate was preferentially utilized by nitrate-reducing bacteria but not by acetate-utilizing SRB.

Biofilms↗
PubMed1998

The evolution of biofilms in venous access devices implanted in children with Wilms' tumour.

Biofilms form on all implanted foreign materials. In venous access ports (VAPs), the biofilm with entrapped organisms may be the source of recurring bacteraemia. At present, little is known of the development of biofilms in VAPs. In this study light, scanning, and transmission electron microscopy were used to investigate the evolution of biofilms in VAPs implanted in 15 African children with Wilms' tumour. The VAPs were removed either emergently because of infection (n = 6) or blockage (n = 3), or electively at the end of chemotherapy (n = 6). Intact biofilms were obtained from lengths of the catheter attached to ports that had been in place for 11 days to 3 years. Each was prepared for light and electron microscopy. In infected ports, shortly after implantation biofilms were thin and comprised of apparently healthy erythrocytes (RBCs) and occasional platelets, leucocytes (WBCs), and bacteria enmeshed in a network of fibrin. Three weeks after implantation, RBCs had autolysed and large numbers of WBCs and bacterial colonies were present within and on the luminal surface. In 1 instance, the lumen of a VAP had been occluded by a "plug" of WBCs. In non-infected patients, the biofilms in long-standing VAPs were of varying thickness and primarily composed of an amorphous granular material. In most cases, healthy and necrotic bacteria were present both within the core and on the surface of the biofilms. The results suggest that while bacteria, per se, are an important factor, the presence and degradation of blood components may be an equally important factor in the development of biofilms in VAP catheters.

Antineoplastic Agents↗
PubMed2002

Minimal attachment killing (MAK): a versatile method for susceptibility testing of attached biofilm-positive and -negative Staphylococcus epidermidis.

Due to its ability to attach to polymeric surfaces Staphylococcus epidermidis is a common pathogen in chronic, medical device-associated infections. Attached S. epidermidis displays reduced susceptibility against a variety of antimicrobial substances, and little correlation between standard susceptibility test results and clinical outcome of antibiotic treatment is observed. In this study we established a new, versatile, and easy method of antimicrobial susceptibility testing for attached Staphylococcus epidermidis, suitable for both biofilm-negative and biofilm-positive attached bacteria using readily available equipment. For three biofilm-positive wild-type strains and their biofilm-negative mutants minimal attachment killing concentrations (MAK) of penicillin, oxacillin, vancomycin, and gentamicin were determined. Depending on strain and investigated antibiotics, a heterogeneous MAK (MAK(hetero)) could be differentiated from a homogeneous resistance (MAK(homo)), favoring a model of few persisters within attached cells under antibiotic treatment. For the biofilm-negative mutants, a lower MAK(homo) was detected than for the corresponding wild types for some of the tested antibiotics, which probably resulted from higher bacterial inocula of wild-type strains, whereas the MAK(hetero) were comparable for mutants and wild types for most of the tested antibiotics and strains. These data indicate that biofilm formation is not a necessary prerequisite for persistence of attached S. epidermidis cells under antibiotic treatment, which could explain therapeutic failure in foreign body-associated infections due to biofilm-negative S. epidermidis isolates. The highly individual resistance phenotypes of the investigated strains with different antibiotics suggests that MAK determination could help to predict the therapeutic outcome of foreign body-associated infections with both biofilm-positive and biofilm-negative S. epidermidis.

Anti-Bacterial Agents↗
PubMed2006

Characterization and performance of constructed nitrifying biofilms during nitrogen bioremediation of a wastewater effluent.

Constructed ammonium oxidizing biofilms (CAOB) and constructed nitrite oxidizing biofilms (CNOB) were characterized during the bioremediation of a wastewater effluent. The maximum ammonium removal rate and removal efficiency in CAOB was 322 mg N-NH4+ m(-3) d(-1) and 96%, respectively, while in CNOB a maximum removal rate of 255 mg N-NH4+ m(-3) d(-1) and a removal efficiency of 76% was achieved. Both constructed biofilms on low-density polyester Dacron support achieved removal efficiencies higher than that of the concentrations normally present in reactors without constructed biofilms (P < 0.05). Nitrifying bacteria from the constructed biofilms cultures were typed by sequencing 16S rRNA genes that had been amplified by PCR from genomic DNA. Analysis of enrichment biofilms has therefore provided evidence of high removal of ammonium and the presence of Nitrosomonas eutropha, N. halophila and N. europaea in CAOB, while in CNOB Nitrobacter hamburgensis, N. winogradskyi and N. alkalicus were identified according to 16S rRNA gene sequences comparison. The biofilm reactors were nitrifying over the whole experimental period (15 days), showing a definite advantage of constructed biofilms for enhancing a high biomass concentration as evidenced by environmental electron microscopic analysis (ESEM). Our research demonstrates that low-density polyester Dacron can be effectively used for the construction of nitrifying biofilms obtaining high removal efficiencies of nitrogen in a relatively short time from municipal effluents from wastewater treatment plants. CAOB and CNOB are potentially promissory for the treatment of industrial wastewaters that otherwise requires very large and expensive reactors for efficient bioremediation of effluents.

Bacteria↗
PubMed1996

Effect of low-intensity ultrasound upon biofilm structure from confocal scanning laser microscopy observation.

Ultrasonic irradiation at 500 kHz and 10 mW cm-2 of a 24 h old biofilm of P. aeruginosa enhanced the killing of bacteria by gentamicin. To determine whether this bioacoustic effect was caused by ultrasonic-induced changes in the biofilm morphology (biofilm breakup or disruption), the biofilms were examined by confocal scanning laser microscopy (CSLM). Such disruption would be undesirable in the possible ultrasonic treatment of implant infections. The CSLM results showed that the biofilm is a partial monolayer of cells with occasional aggregates of cells, non-cellular materials and extracellular spaces. The aggregates contained large amounts of exopolysaccharide. The structure of biofilm was not changed when the biofilm was exposed to continuous ultrasound at 500 kHz and 10 mW cm-2, the same irradiation parameters that increased cell killing by nearly two orders of magnitude. The observation that low-intensity ultrasound does not disrupt biofilm or disperse the bacteria has significance in the possible use of ultrasound to enhance the action of antibiotics against biofilms.

Acoustics↗
PubMed2005

Biofilms and antibiotic therapy: is there a role for combating bacterial resistance by the use of novel drug delivery systems?

The conventional view of antibiotic resistance is one where bacteria exhibit significantly reduced susceptibility to antimicrobials in laboratory tests by mechanisms such as altered drug uptake, altered drug target and drug inactivation. Whilst these mechanisms undoubtedly make a major contribution to antibiotic failure in the clinic, the phenomenon of clinical failure in spite of sensitivity in laboratory tests is also well recognised. It is in this context that attention has focussed on bacteria growing as adherent biofilms, not only as the mode of growth of device-related infections associated for example with artificial joints and venous catheters, but also with other chronic infections such as those occurring in the respiratory tract. Growth as a biofilm almost always leads to a significant decrease in susceptibility to antimicrobial agents compared with cultures grown in suspension and, whilst there is no generally agreed mechanism for the resistance of biofilm bacteria, it is largely phenotypic. That is, when biofilm bacteria are grown in conventional laboratory suspension culture they become susceptible to antimicrobials. A number of elements in the process of biofilm formation have been studied as targets for novel drug delivery technologies. These include surface modification of devices to reduce bacterial attachment and biofilm development as well as incorporation of antimicrobials-again to prevent colonisation. Electrical approaches have been used either to release antimicrobials from device surfaces or to drive antimicrobials through the biofilm. Other technologies not specifically focussed on biofilms include aerosolized delivery of antibiotics to the lung and formulation into liposome and polymer-based vehicles. Liposomal systems have been widely studied, either to target antibiotics to the surface of bacterial biofilms, or by virtue of their property of being taken up cells of the reticuloendothelial system, to target antibiotics towards intracellular bacteria. Many polymer-based carrier systems have also been proposed, including those based on biodegradable polymers such as poly(lactide-co-glycolide) as well as thermoreversible hydrogels. Their contribution to the prevention or resolution of infection is reviewed.

Anti-Bacterial Agents↗
PubMed2005

Prevalence of Staphylococcus epidermidis strains with biofilm-forming ability in isolates from conjunctiva and facial skin.

PURPOSE: To compare the prevalence of biofilm-forming strains of Staphylococcus epidermidis in the conjunctival and facial skin microflora. DESIGN: Experimental study. METHODS: The prevalence of biofilm-forming ability of 10 S epidermidis strains obtained from the conjunctival sac of healthy volunteers was compared with 40 strains obtained from the facial skin of healthy volunteers. The ability to form biofilm was determined by the presence of the icaA gene, and the production of biofilm was examined by qualitative (Congo red agar [CRA]) and quantitative (microtiter plate) assays. Additionally, the prevalence of 36 S epidermidis strains obtained from the conjunctival sac of precataract patients to form biofilm was investigated. RESULTS: The icaA gene was detected in 60% of the isolates from the conjunctival sac of volunteers and 15% of those from the facial skin. Fifty percent of the isolates from the conjunctiva of volunteers and 5% from the facial skin were CRA positive. Biofilm production was significantly greater in isolates from the conjunctiva of volunteers. Of the nine pairs of isolates found in the same volunteers, six conjunctival sac isolates were positive for the icaA gene with biofilm-forming ability except one strain, whereas only one of the facial skin isolates was positive for the icaA gene and none exhibited biofilm-forming phenotype. Sixty-nine percent and 44% of the isolates from the conjunctival sac of precataract patients were positive for icaA gene and CRA test, respectively. CONCLUSIONS: The prevalence of biofilm-forming S epidermidis isolates is higher in the conjunctival sac than the facial skin.

Aged↗
PubMed2005

Investigating the suitability of the Calgary Biofilm Device for assessing the antimicrobial efficacy of new agents.

This study investigated the suitability of the Calgary Biofilm Device (CBD), originally designed as a test surrogate for indwelling medical devices, for assessing the efficacy of antimicrobials developed for food and food contact surface disinfection applications. The conditions for the development of uniform biofilms from pure and mixed bacterial cultures of wild type Escherichia coli and Listeria innocua were optimized. We were able to recover approximately 2 x 10(6) colony forming units (CFU) from the biofilms formed on the individual pegs of the device in 24 h. Further, the parameters for the consistent release of the cells from the biofilms were optimized; test showed that the number of cells released was uniform and reproducible. The consistency and reproducibility of the biofilms formed on the pegs was evaluated using scanning electron microscopy and by plate count method. The efficacies of disinfectants on cells residing in biofilms versus planktonic cells were compared. For both species, higher concentrations of disinfectants were needed to eliminate attached cells as compared with planktonic cells. This study establishes the value of the CBD for generating consistent biofilms from either pure or mixed cultures. These biofilms can be used to assess efficacies of disinfectants against cells that have colonized the surfaces of foods and food-processing equipment. Such a system could serve as a standard surrogate for evaluating new disinfectants designed to reduce or eliminate biofilms from food-contact surfaces.

Anti-Infective Agents↗
PubMedDate not supplied

Candida biofilm resistance.

Device-related infections in most nosocomial diseases can be traced to the formation of biofilms (microbial communities encased within polysaccharide-rich extracellular matrix) by pathogens on surfaces of these devices. Candida species are the most common fungi isolated from these infections, and biofilms formed by these fungal organisms are associated with drastically enhanced resistance against most antimicrobial agents. This enhanced resistance contributes to the persistence of this fungus despite antifungal therapy. Candida biofilms exhibit enhanced resistance against most antifungal agents, except echinocandins and lipid formulations of AmB. The expression of drug efflux pumps during the early phase of biofilm formation and alterations in membrane sterol composition contribute to resistance of these biofilms against azoles. Metabolic dormancy and ECM do not appear to contribute to resistance, although in a mixed-species biofilm, ECM does retard the diffusion of drugs across biofilm. These multifactorial mechanisms of resistance in fungal biofilms constitute a broad-spectrum defense that is effective against many types of antifungal agents, and represent a common theme present across microbial biofilms.

Animals↗