Crystalline Biofilms Research Articles

Bacteriophage Can Prevent Encrustation and Blockage of Urinary Catheters by Proteus mirabilis.

Proteus mirabilis forms dense crystalline biofilms on catheter surfaces that occlude urine flow, leading to serious clinical complications in long-term catheterized patients, but there are presently no truly effective approaches to control catheter blockage by this organism. This study evaluated the potential for bacteriophage therapy to control P. mirabilis infection and prevent catheter blockage. Representative in vitro models of the catheterized urinary tract, simulating a complete closed drainage system as used in clinical practice, were employed to evaluate the performance of phage therapy in preventing blockage. Models mimicking either an established infection or early colonization of the catheterized urinary tract were treated with a single dose of a 3-phage cocktail, and the impact on time taken for catheters to block, as well as levels of crystalline biofilm formation, was measured. In models of established infection, phage treatment significantly increased time taken for catheters to block (∼ 3-fold) compared to untreated controls. However, in models simulating early-stage infection, phage treatment eradicated P. mirabilis and prevented blockage entirely. Analysis of catheters from models of established infection 10 h after phage application demonstrated that phage significantly reduced crystalline biofilm formation but did not significantly reduce the level of planktonic cells in the residual bladder urine. Taken together, these results show that bacteriophage constitute a promising strategy for the prevention of catheter blockage but that methods to deliver phage in sufficient numbers and within a key therapeutic window (early infection) will also be important to the successful application of phage to this problem.

Novel Insights into the Proteus mirabilis Crystalline Biofilm Using Real-Time Imaging.

The long-term use of indwelling catheters results in a high risk from urinary tract infections (UTI) and blockage. Blockages often occur from crystalline deposits, formed as the pH rises due to the action of urease-producing bacteria; the most commonly found species being Proteus mirabilis. These crystalline biofilms have been found to develop on all catheter materials with P. mirabilis attaching to all surfaces and forming encrustations. Previous studies have mainly relied on electron microscopy to describe this process but there remains a lack of understanding into the stages of biofilm formation. Using an advanced light microscopy technique, episcopic differential interference contrast (EDIC) microscopy combined with epifluorescence (EF), we describe a non-destructive, non-contact, real-time imaging method used to track all stages of biofilm development from initial single cell attachment to complex crystalline biofilm formation. Using a simple six-well plate system, attachment of P. mirabilis (in artificial urine) to sections of silicone and hydrogel latex catheters was tracked over time (up to 24 days). Using EDIC and EF we show how initial attachment occurred in less than 1 h following exposure to P. mirabilis. This was rapidly followed by an accumulation of an additional material (indicated to be carbohydrate based using lectin staining) and the presence of highly elongated, motile cells. After 24 h exposure, a layer developed above this conditioning film and within 4 days the entire surface (of both catheter materials) was covered with diffuse crystalline deposits with defined crystals embedded. Using three-dimensional image reconstruction software, cells of P. mirabilis were seen covering the crystal surfaces. EDIC microscopy could resolve these four components of the complex crystalline biofilm and the close relationship between P. mirabilis and the crystals. This real-time imaging technique permits study of this complex biofilm development with no risk of artefacts due to sample manipulation. A full understanding of the stages and components involved in crystalline encrustation formation will aid in the development of new protocols to manage and ultimately prevent catheter blockage.

Irrigation with N,N-dichloro-2,2-dimethyltaurine (NVC-422) in a citrate buffer maintains urinary catheter patency in vitro and prevents encrustation by Proteus mirabilis.

Long-term use of indwelling urinary catheters can lead to urinary tract infections and loss of catheter patency due to encrustation and blockage. Encrustation of urinary catheters is due to formation of crystalline biofilms by urease-producing microorganisms such as Proteus mirabilis. An in vitro catheter biofilm model (CBM) was used to evaluate current methods for maintaining urinary catheter patency. We compared antimicrobial-coated urinary Foley catheters, with both available catheter irrigation solutions and investigational solutions containing NVC-422 (N,N-dichloro-2,2-dimethyltaurine; a novel broad-spectrum antimicrobial). Inoculation of the CBM reactor with 10(8) colony-forming units of P. mirabilis resulted in crystalline biofilm formation in catheters by 48h and blockage of catheters within 5days. Silver hydrogel or nitrofurazone-coated catheters did not extend the duration of catheter patency. Catheters irrigated daily with commercially available solutions such as 0.25% acetic acid and isotonic saline blocked at the same rate as untreated catheters. Daily irrigations of catheters with 0.2% NVC-422 in 10mM acetate-buffered saline pH 4 or Renacidin maintained catheter patency throughout 10-day studies, but P. mirabilis colonization of the CBM remained. In contrast, 0.2% NVC-422 in citrate buffer (6.6% citric acid at pH 3.8) resulted in an irrigation solution that not only maintained catheter patency for 10days but also completely eradicated the P. mirabilis biofilm within one treatment day. These data suggest that an irrigation solution containing the rapidly bactericidal antimicrobial NVC-422 in combination with citric acid to permeabilize crystalline biofilm may significantly enhance catheter patency versus other approved irrigation solutions and antimicrobial-coated catheters.

Hydrogenated amorphous carbon coatings on implants drastically reduce biofilm formation and water permeation

Inflammations and crystalline bacterial biofilms (encrustations) remain a major complication in long-term artificial urinary tract drainage. To solve this problem we present urological implants with coatings made of amorphous hydrogenated carbon (a-C:H) that show excellent protection from encrustation in-vitro as well as in-vivo. Part of the success of a-C:H coatings is attributed to their ability to act as a diffusion barrier between an implant and the body, which prevents leaching of solvents from polymeric implants. To further enhance their barrier properties a-C:H coatings are combined with parylene coatings to develop diffusion-barrier multilayer coatings with a total thickness between 0.2 μm and 0.8 μm. The combination of the two types of coatings leads to a reduction of water diffusion by a factor of up to ten with respect to uncoated 25 μm thick polyimide sub-strates. The diffusion of water vapour from a controlled atmospheric pressure chamber through coated foils to a vacuum chamber is measured in a custom-built device.

Clinical complications of urinary catheters caused by crystalline biofilms: something needs to be done

This review is largely based on a previous paper published in the journal Spinal Cord. The care of many patients undergoing long-term bladder catheterization is complicated by encrustation and blockage of their Foley catheters. This problem stems from infection by urease-producing bacteria, particularly Proteus mirabilis. These organisms colonize the catheter forming an extensive biofilm; they also generate ammonia from urea, thus elevating the pH of urine. As the pH rises, crystals of calcium and magnesium phosphates precipitate in the urine and in the catheter biofilm. The continued development of this crystalline biofilm blocks the flow of urine through the catheter. Urine then either leaks along the outside of the catheter and the patient becomes incontinent or is retained causing painful distension of the bladder and reflux of urine to the kidneys. The process of crystal deposition can also initiate stone formation. Most patients suffering from recurrent catheter encrustation develop bladder stones. P.mirabilis establishes stable residence in these stones and is extremely difficult to eliminate from the catheterized urinary tract by antibiotic therapy. If blocked catheters are not identified and changed, serious symptomatic episodes of pyelonephritis, septicaemia and endotoxic shock can result. All types of Foley catheters including silver- or nitrofurazone-coated devices are vulnerable to this problem. In this review, the ways in which biofilm formation on Foley catheters is initiated by P.mirabilis will be described. The implications of understanding these mechanisms for the development of an encrustation-resistant catheter will be discussed. Finally, the way forward for the prevention and control of this problem will be considered.

MP20-01 AURICLOSENE IRRIGATION SOLUTION PREVENTS ENCRUSTATION BY CRYSTALLINE BIOFILM DUE TO PROTEUS MIRABILIS IN AN IN VITRO URINARY CATHETER PATENCY MODEL

INTRODUCTION AND OBJECTIVES: Long-term indwelling urinary catheters are susceptible to blockage due to formation of crystalline biofilms by urease-producing microorganisms such as Proteus mirabilis. An in vitro catheter biofilm model (CBM) was developed to compare current methods for maintaining urinary catheter patency. We compared various bladder irrigation solutions or antimicrobial-coated urinary catheters, versus a novel anti-microbial catheter irrigation solution containing auriclosene (N,N-dichloro-2,2-dimethyltaurine; formerly designated NVC-422). METHODS: CBM units were fed artificial urine at 0.5 mL per min. The artificial bladder chamber was inoculated with 10 8 colony forming units (CFU) of P. mirabilis and biofilm was allowed to establish for 48 hours before daily treatments commenced. A single treatment consisted of two sequential 50-mL irrigations. Each irrigation was retained in the catheter for 15 min and then drained, with a 30-min washout period between the two irrigations. Experiments were conducted for up to 10 days or until catheter blockage. The pH of the effluent, CFU counts in the bladder chamber and the time to catheter blockage were recorded. The area of catheter encrustation was measured using Stereo Zoom imaging. RESULTS: Inoculation of the CBM reactor with 10 8 CFU of P. mirabilis resulted in blockage of the urinary catheters within 5 days. The use of silver-hydrogel or nitrofurazone-coated catheters did not extend the period of catheter patency. Catheters irrigated with 0.25% acetic acid, 10 mM acetate-buffered saline or isotonic saline blocked at the same rate as untreated catheters. Catheter irrigation with a citrate-buffered formulation of 0.2% auriclosene resulted in complete eradication of P. mirabilis biofilm within one treatment day. In contrast, daily irrigations of infected catheters with 0.2% auriclosene in 10 mM acetate-buffered saline (at pH 4) or Renacidin ® Irrigation Solution had no effect on P. mirabilis colonization of the bladder chamber, even though catheter patency was maintained throughout 10-day studies. CONCLUSIONS: Irrigation with the rapidly bactericidal antimicrobial auriclosene in a buffered acidic formulation - termed Auriclosene Irrigation Solution - significantly enhanced catheter patency in vitro versus other irrigation solutions and antimicrobial- coated urinary catheters. Clinical evaluation of Auriclosene Irrigation Solution is ongoing. Results (continued) Abstract

Soft Robotic Concepts in Catheter Design: an On‐Demand Fouling‐Release Urinary Catheter

Infectious biofilms are problematic in many healthcare-related devices and are especially challenging and ubiquitous in urinary catheters. This report presents an on-demand fouling-release methodology to mechanically disrupt and remove biofilms, and proposes this method for the active removal of infectious biofilms from the previously inaccessible main drainage lumen of urinary catheters. Mature Proteus mirabilis crystalline biofilms detach from silicone elastomer substrates upon application of strain to the substrate, and increasing the strain rate increases biofilm detachment. The study presents a quantitative relationship between applied strain rate and biofilm debonding through an analysis of biofilm segment length and the driving force for debonding. Based on this mechanism, hydraulic and pneumatic elastomer actuation is used to achieve surface strain selectively within the lumen of prototypes of sections of a fouling-release urinary catheter. Proof-of-concept prototypes of sections of active, fouling-release catheters are constructed using techniques typical to soft robotics including 3D printing and replica molding, and those prototypes demonstrate release of mature P. mirabilis crystalline biofilms (e.g., ≈90%) from strained surfaces. These results provide a basis for the development of a new urinary catheter technology in which infectious biofilms are effectively managed through new methods that are entirely complementary to existing approaches.

A clinical evaluation of a sensor to detect blockage due to crystalline biofilm formation on indwelling urinary catheters

To test the performance and acceptability of an early warning sensor to predict encrustation and blockage of long-term indwelling urinary catheters. In all, 17 long-term indwelling catheter users, 15 'blockers' and two 'non-blockers' (controls) were recruited; 11 participants were followed prospectively until catheter change, three withdrew early and three did not start. Two sensors were placed in series between the catheter and the urine bag at catheter change. The sensor nearest the bag was changed at the same time as the bag change (weekly); the sensor nearest the catheter remained in situ for the duration of the catheter's life. Bacteriology and pH determinations were performed on urine samples at each bag, sensor and catheter change. The colour of the sensors was recorded daily. On removal, each sensor and the catheter were examined for visible evidence of encrustation and blockage. Participants were asked to keep a daily diary to record colour change and any other relevant observations and to complete a psychosocial impact of assistive devices tool at the end of the study. Participants and carers/healthcare professionals (when involved in urine bag or catheter change) were asked to complete a questionnaire about the sensor. Urease-producing bacteria were isolated from seven of the 14 patients (including early withdrawals; P. mirabilis in four, Morganella or Providencia in three). In six of the seven patients the sensors turned blue-black; two of these were early withdrawals, two went to planned catheter change (one of these was recruited as a 'non-blocker') and three had catheter blockage. The number of days of catheterisation before blockage was 22, 23 and 25 days, and the sensor changed colour within 24-48 h after insertion. The urine mean (range) pH of the sensors that turned blue-black was 7.6 (5.5-9.0) and of the sensors that remained yellow 6.1 (5.1-7.5). The sensor was generally well-received and was positive in the psychosocial assessment. The sensor is a useful indicator of urine pH and of the conditions that lead to catheter blockage. It may be particularly useful for new indwelling catheter users. To be a universally acceptable predictor of catheter blockage, the time from sensor colour change to blockage needs to be reduced.

Elucidating the Genetic Basis of Crystalline Biofilm Formation in Proteus mirabilis

Proteus mirabilis forms extensive crystalline biofilms on urethral catheters that occlude urine flow and frequently complicate the management of long-term-catheterized patients. Here, using random transposon mutagenesis in conjunction with in vitro models of the catheterized urinary tract, we elucidate the mechanisms underpinning the formation of crystalline biofilms by P. mirabilis. Mutants identified as defective in blockage of urethral catheters had disruptions in genes involved in nitrogen metabolism and efflux systems but were unaffected in general growth, survival in bladder model systems, or the ability to elevate urinary pH. Imaging of biofilms directly on catheter surfaces, along with quantification of levels of encrustation and biomass, confirmed that the mutants were attenuated specifically in the ability to form crystalline biofilms compared with that of the wild type. However, the biofilm-deficient phenotype of these mutants was not due to deficiencies in attachment to catheter biomaterials, and defects in later stages of biofilm development were indicated. For one blocking-deficient mutant, the disrupted gene (encoding a putative multidrug efflux pump) was also found to be associated with susceptibility to fosfomycin, and loss of this system or general inhibition of efflux pumps increased sensitivity to this antibiotic. Furthermore, homologues of this system were found to be widely distributed among other common pathogens of the catheterized urinary tract. Overall, our findings provide fundamental new insight into crystalline biofilm formation by P. mirabilis, including the link between biofilm formation and antibiotic resistance in this organism, and indicate a potential role for efflux pump inhibitors in the treatment or prevention of P. mirabilis crystalline biofilms.

Prevention of Proteus mirabilis Biofilm by Surfactant Solution

Background: The crystalline biofilms formed by Proteus mirabilis can seriously complicate the care of patients undergoing bladder catheterization, the prevention of crystalline biofilms is important to avoid urinary catheter complications. The surfactant of Lactobacillus acidophilus can be interacting with attachment of many microorganisms. Aims of the study: 1- To detect the biofilm formation ability of Proteus mirabilis isolates in urinary catheter. 2- To detect the growth and biofilm formation ability of Proteus mirabilis isolates on 96-wells microtiter plates. 3- To study the role of Lactobacillus acidophilus surfactant in biofilm formed by Proteus mirabilis isolates. Materials and methods: Since Jan/2011 to Jan/2012 a 48 isolates of Proteus mirabilis were isolated from the encrusted catheter of a patient undergoing indwelling catheterization in Al-Ramadi General Hospital while Lactobacillus acidophilus isolate was obtained from urogenital tracts of healthy woman by vaginal swab and surfactant was extracted from it. The biofilm formation on catheters, quantitative assays of biofilm formation and biofilm inhibition assay by surfactant were studied. Results: Fourty five 45 (93.75%) P. mirabilis isolates produced a biofilm in urinary catheter while 3 (6.25%) isolates were not produce biofilm while 48 (100%) isolates forms good biofilms results on 96-wells microtiter plate, in brain heart infusion broth with 0.2% glucose, and at 37°C. The growth of P. mirabilis isolates in 96-well microtiter plates were determine at OD630, the optimum growth of P. mirabilis isolates were recorded after 48 hours of incubation (OD630= 0.65). Biofilm kinetics of P. mirabilis isolates refers to that attached cells increased with time and the maximum biofilm formation ratio occurs at 48 hours of incubation (OD550= 0.70). Surfactant show a good ability to inhibit biofilm formation, the increasing amounts of surfactant led to a decrease in the amount of biofilm formed by P. mirabilis isolates and that 6.0 µg/ml of surfactant was more than sufficient to completely abolish biofilm formation.

Iron oxide and calcite associated withLeptothrixsp. biofilms within an estavelle in the upper Floridan aquifer

Florea L.J., Noe-Stinson C.L., Brewer J., Fowler R., Kearns B.J. and Greco A.M. 2011. Iron oxide and calcite associated with Leptothrix sp. biofilms within an estavelle in the upper Floridan aquifer. International Journal of Speleology, 40 (2), 205-219. Tampa, FL (USA). ISSN 0392-6672. DOI 10.5038/1827-806X.40.2.12 In Thornton’s Cave, an estavelle in west-central Florida, SEM, EDS, and XRD data reveal biofilms that are predominantly comprised of FeOOH-encrusted hollow sheaths that are overgrown and intercalated with calcite. Fragments of this crystalline biofilm adhere to the walls and ceiling as water levels vary within the cave. Those on the wall have a ‘cornflake’ appearance and those affixed to the ceiling hang as fibrous membranes. PCR of DNA in the active biofilm, combined with morphologic data from the tubes in SEM micrographs, point to Leptothrix sp., a common Fe-oxidizing bacteria, as the primary organism in the biofilm. Recent discoveries of ‘rusticles’ in other Florida caves suggest that Fe-oxidizing bacteria may reside elsewhere in Florida groundwater and may play a role in the mobility of trace metals in the Upper Florida aquifer. SEM micrographs from two marble tablets submerged for five months, one exposed to microbial activity and a second isolated from microbial action, revealed no visible etchings or borings and very limited loss of mass. EDS data from the electron micrographs of the unfiltered tablet document the same FeOOH-encrusted hollow sheaths and similar deposits of calcite as seen in the ‘cornflakes’. These results, combined with water chemistry data imply that the biofilm may focus or even promote calcite precipitation during low-water level conditions when CO 2 degasses from the cave pools.

A three-phase in-vitro system for studying Pseudomonas aeruginosa adhesion and biofilm formation upon hydrogel contact lenses

BackgroundPseudomonas aeruginosa is commonly associated with contact lens (CL) -related eye infections, for which bacterial adhesion and biofilm formation upon hydrogel CLs is a specific risk factor. Whilst P. aeruginosa has been widely used as a model organism for initial biofilm formation on CLs, in-vitro models that closely reproduce in-vivo conditions have rarely been presented.ResultsIn the current investigation, a novel in-vitro biofilm model for studying the adherence of P. aeruginosa to hydrogel CLs was established. Nutritional and interfacial conditions similar to those in the eye of a CL wearer were created through the involvement of a solid:liquid and a solid:air interface, shear forces and a complex artificial tear fluid. Bioburdens varied depending on the CL material and biofilm maturation occurred after 72 h incubation. Whilst a range of biofilm morphologies were visualised including dispersed and adherent bacterial cells, aggregates and colonies embedded in extracellular polymer substances (EPS), EPS fibres, mushroom-like formations, and crystalline structures, a compact and heterogeneous biofilm morphology predominated on all CL materials.ConclusionsIn order to better understand the process of biofilm formation on CLs and to test the efficacy of CL care solutions, representative in-vitro biofilm models are required. Here, we present a three-phase biofilm model that simulates the environment in the eye of a CL wearer and thus generates biofilms which resemble those commonly observed in-situ.

Silver or Nitrofurazone Impregnation of Urinary Catheters Has a Minimal Effect on Uropathogen Adherence

Bacterial adherence to the urinary catheter is an early step in biofilm formation and the pathogenesis of catheter associated urinary tract infection. We studied in vitro the effect of silver or nitrofurazone impregnation of urinary catheters on uropathogen ability to adhere to urinary catheters. We studied commercially available nitrofurazone-silicone, silicone only, silver-silicone-hydrogel, silicone-hydrogel, silver-latex-hydrogel and latex-hydrogel catheters. Catheters were incubated in sterile broth for 0, 3, 5, 7 and 10 days, respectively, before inoculation and overnight incubation with Escherichia coli or Enterococcus faecalis. Adherence of E. coli and E. faecalis to nitrofurazone catheters was significantly decreased compared to that of silicone-only catheters when catheters were fresh. The anti-adherence effect of nitrofurazone on E. coli decreased with time but was still significant at 5 days. For E. faecalis the effect of nitrofurazone was lost by 3 days of pre-incubation. E. coli adherence was not significantly decreased on silver impregnated catheters compared to that on control catheters of the same base material. Silver was associated with a significant decrease in E. faecalis adherence to latex-hydrogel catheters but not to silicone-hydrogel catheters. The adherence of each species to silicone catheters with hydrogel was significantly lower than that to silicone-only control catheters. Silver impregnation had little effect on bacterial adherence in our model and nitrofurazone impregnation had a significant effect only for the first 5 days. Our results do not support a role for silver urinary catheters to prevent catheter associated urinary tract infection by decreasing bacterial adherence.

In Vitro Multicompartmental Bladder Model for Assessing Blockage of Urinary Catheters: Effect of Hydrogel Coating on Dynamics of Proteus mirabilis Growth

To investigate the effect of a hydrogel coating on the dynamics of bacterial growth in laboratory models of the catheterized bladder. Infection of the urinary tract by Proteus mirabilis can result in catheter blockage by crystalline biofilm, a common complication in patients undergoing long-term bladder catheterization. Two series of catheters were tested in the infected bladder models: test series 1, silicone catheters impregnated with triclosan (0.5%, 1%, 4%), or silicone catheters with 0% triclosan impregnated with pure solvents and hydrogel coated (based on polyvinylpyrrolidone); and test series 2, silicone catheters, hydrogel-coated with hydrogel plus iodine (polyvinylpyrrolidone plus iodine) or hydrogel plus polyhexamethylene biguanide. Test series 1 was used to detect the influence of triclosan, solvents, impregnation time, and the presence of hydrogel coating on the interval to catheter blockage by P. mirabilis biofilm. The experiments with test series 2 focused on the dynamic interaction of the hydrogel coating and biofilm formation. The division of the catheterized bladder model into 3 sampling zones brought more information about the spatial segregation of the bacterial population. The bacteriostatic efficiency of the water-soluble polyhexamethylene biguanide and polyvinylpyrrolidone iodine complex was limited to the first hours after catheterization. Only catheters containing triclosan resisted encrustation for significantly longer (up to >7 days). In contrast, the uncoated and hydrogel-coated catheters were occluded by day 2. The hydrogel layer can increase aggregation of the planktonic cells and newly nucleated crystals, leading to even faster catheter blockage than in the case of uncoated silicone. However, the addition of active agents were able to suppress this negative effect.

The encrustation and blockage of long-term indwelling bladder catheters: a way forward in prevention and control

To review the literature showing that understanding how Foley catheters become encrusted and blocked by crystalline bacterial biofilms has led to strategies for the control of this complication in the care of patients undergoing long-term indwelling bladder catheterization. A comprehensive PubMed search of the literature published between 1980 and December 2009 was made for relevant articles using the Medical Subject Heading terms 'biofilms', 'urinary catheterization', 'catheter-associated urinary tract infection' and 'urolithiasis'. Papers on catheter-associated urinary tract infections and bacterial biofilms collected during 40 years of working in the field were also reviewed. There is strong experimental and epidemiological evidence that infection by Proteus mirabilis is the main cause of the crystalline biofilms that encrust and block Foley catheters. The ability of P. mirabilis to generate alkaline urine and to colonize all available types of indwelling catheters allows it to take up stable residence in the catheterized tract in bladder stones and cause recurrent catheter blockage. The elimination of P. mirabilis by antibiotic therapy as soon as it appears in the catheterized urinary tract could improve the quality of life for many patients and reduce the current expenditure of resources when managing the complications of catheter encrustation and blockage. For patients who are already chronic blockers and stone formers, antibiotic treatment is unlikely to be effective owing to the resistance of cells in the crystalline biofilms. Strategies such as increasing fluid intake with citrated drinks could control the problem until bladder stone removal can be organized.

Crystalline bacterial biofilm formation on urinary catheters by urease-producing urinary tract pathogens: a simple method of control

The problem of catheter encrustation stems from infection by urease-producing bacteria. These organisms generate ammonia from urea, elevate the pH of urine and cause crystals of calcium and magnesium phosphates to form in the urine and the biofilm that develops on the catheter. In this study, a laboratory model was used to compare the ability of 12 urease-positive species of urinary tract pathogens to encrust and block catheters. Proteus mirabilis, Proteus vulgaris and Providencia rettgeri were able to raise the urinary pH above 8.3 and produce catheter-blocking crystalline biofilms within 40 h. Morganella morganii and Staphylococcus aureus elevated the pH of urine to 7.4 and 6.9, respectively, and caused some crystal deposition in the biofilms but did not block catheters in the 96 h experimental period. Isolates of Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, Serratia marcescens, Pseudomonas aeruginosa and Providencia stuartii were only capable of raising the pH of urine to a maximum of 6.4 and failed to cause crystal deposition in the biofilm. The most effective way to prevent catheter encrustation was shown to be diluting urine and increasing its citrate concentration. This strategy raises the nucleation pH (pH(n)) at which calcium and magnesium phosphates crystallize from urine. Increasing the fluid intake of a healthy volunteer with citrated drinks resulted in urine with a pH(n) of >8.0 in which catheter encrustation was inhibited. It is suggested that this dietary strategy will be an effective means of controlling catheter encrustation, whichever bacterial species is causing the problem.

The effect of EDTA instillations on the rate of development of encrustation and biofilms in Foley catheters

The aim of this research was to examine whether a daily instillation of tetra sodium ethylenediaminetetraacetic acid (EDTA) solution could reduce the rate at which encrustation by crystalline Proteus mirabilis biofilms blocks urinary catheters. Sets of three bladder models were fitted with size 14 all-silicone catheters. Tetra sodium EDTA solution was instilled into the catheter following biofilm development. Catheters were examined by digital photography and scanning electron microscopy for evidence of encrustation. The results showed that the mean time to blockage of the control catheters was 45 h for saline, 57 h for water and 67 h for those exposed to daily instillations of the EDTA solution. Statistical analysis confirmed that the mean encrustation rate on the EDTA-treated catheters was significantly lower than on the control-treated devices (P = 0.047). This in vitro study indicates that EDTA may have beneficial effects in reducing the complication of catheter encrustation and blockage by crystalline biofilms.

The high-affinity phosphate transporter Pst in Proteus mirabilis HI4320 and its importance in biofilm formation

Proteus mirabilis causes urinary tract infections (UTIs) in individuals requiring long-term indwelling catheterization. The pathogenesis of this uropathogen is mediated by a number of virulence factors and the formation of crystalline biofilms. In addition, micro-organisms have evolved complex systems for the acquisition of nutrients, including the phosphate-specific transport system, which has been shown to be important in biofilm formation and pathogenesis. A functional Pst system is important during UTIs caused by P. mirabilis HI4320, since transposon mutants in the PstS periplasmic binding protein and the PstA permease protein were attenuated in the CBA mouse model of UTI. These mutants displayed a defect in biofilm formation when grown in human urine. This study focuses on a comparison of the proteomes during biofilm and planktonic growth in phosphate-rich medium and human urine, and microscopic investigations of biofilms formed by the pst mutants. Our data suggest that (i) the Deltapst mutants, and particularly the DeltapstS mutant, are defective in biofilm formation, and (ii) the proteomes of these mutants differ significantly from that of the wild-type. Therefore, since the Pst system of P. mirabilis HI4320 negatively regulates biofilm formation, this system is important for the pathogenesis of these organisms during complicated UTIs.

Reduction of Biofilm Formation on a‐C:H Coated Implants: Investigation of Biofilm‐Surface Interactions by Variation of Thin Film Properties

AbstractFormation of crystalline bacterial biofilms (encrustations) on urological implants remains a major complication in patient care. Recently, it was shown that amorphous carbon (a‐C:H) coatings deposited on polyurethane (PU) stents provide a distinct reduction in biofilm formation with an effect more pronounced in‐vivo than in‐vitro. To investigate the underlying mechanisms, the influence of surface properties on the encrustation tendency of plasma coated and uncoated PU tubes was tested in an extended in‐vitro model with protein‐spiked artificial urine and urease. In the presence of albumin the formation of hollow crystalline spheres was observed mainly on uncoated PU. We suggest that evolving gas bubbles influence protein adherence and attachment of crystalline bacterial biofilm to the implant surfaces.

Impact of wastewater fouling on contact angle

Capillary dependent systems are highly influenced by surface fouling and may degrade as material surface properties change. In anticipation of a spacecraft microgravity fluids management system exposed to highly variable wetting conditions, the impact of urine wastewater fouling on capillary contact angle was examined. The results indicate that, in general, surface fouling can decrease the contact angle when crystalline structures or biofilms form. Small crystalline growth on the order of 10 μm can lower advancing contact angles θadv by approximately 30°, while biofilm growth can lower it by approximately 15°. Vacuum drying of fouled surfaces increased θadv by about 8°, and defects greater in height than 5% of the capillary length increased θadv by approximately 30°. These trends may indicate that promotion of wastewater fouling may improve the performance of capillary dependent fluids management systems. These results may also influence terrestrial technologies, including medical catheters and sustainable wastewater treatment systems exposed to wastewater fouling.