Role of swarming in the formation of crystalline Proteus mirabilis biofilms on urinary catheters.

The care of many patients undergoing long-term bladder catheterization is frequently complicated by infection with Proteus mirabilis. These organisms colonize the catheter, forming surface biofilm communities, and their urease activity generates alkaline conditions under which crystals of magnesium ammonium phosphate and calcium phosphate are formed and become trapped in the biofilm. As the biofilm develops it obstructs the flow of urine through the catheter, causing either incontinence due to leakage of urine around the catheter or retention of urine in the bladder. The aim of this study was to investigate the role of the surface-associated swarming motility of P. mirabilis in the initiation and development of these crystalline catheter biofilms. A set of stable transposon mutants with a range of swimming and swarming abilities were tested for their ability to colonize silicone surfaces in a parallel-plate flow cell. A laboratory model of the catheterized bladder was then used to examine their ability to form crystalline, catheter-blocking biofilms. The results showed that neither swarming nor swimming motility was required for the attachment of P. mirabilis to silicone. Mutants deficient in swarming and swimming were also capable of forming crystalline biofilms and blocking catheters more rapidly than the wild-type strain.

Does the valve regulated release of urine from the bladder decrease encrustation and blockage of indwelling catheters by crystalline proteus mirabilis biofilms?

PURPOSE: We tested whether valve regulated, intermittent flow of urine from catheterized bladders decreases catheter encrustation. MATERIALS AND METHODS: Laboratory models of the catheterized bladder were infected with Proteus mirabilis. Urine was allowed to drain continuously through the catheters or regulated by valves to drain intermittently at predetermined intervals. The time that catheters required to become blocked was recorded and encrustation was visualized by scanning electron microscopy. RESULTS: When a manual valve was used to drain urine from the bladder at 2-hour intervals 4 times during the day, catheters required significantly longer to become blocked than those on continuous drainage (mean 62.6 vs 35.9 hours, p = 0.039). A similar 1.7-fold increase occurred when urine was drained at 4-hour intervals 3 times daily. Experiments with an automatic valve in which urine was released at 2 or 4-hour intervals through the day and night also showed a significant increase in mean time to blockage compared with continuous drainage (p = 0.001). Scanning electron microscopy confirmed that crystalline biofilm was less extensive on valve regulated catheters. CONCLUSIONS: Valve regulated, intermittent flow of urine through catheters increases the time that catheters require to become blocked with crystalline biofilm. The most beneficial effect was recorded when urine was released from the bladder at 4-hour intervals throughout the day and night by an automatic valve.

Genotyping demonstrates that the strains of Proteus mirabilis from bladder stones and catheter encrustations of patients undergoing long-term bladder catheterization are identical.

PURPOSE: We established the incidence of bladder stones in patients who experienced recurrent encrustation and blockage of indwelling bladder catheters and examined the relationship between isolates of Proteus mirabilis from the stones and from the crystalline biofilms on the catheters. MATERIALS AND METHODS: The first 100 patients attending a clinic for patients experiencing problems with the management of long-term bladder catheters were studied. Flexible cystoscopy was used to detect bladder stones. Catheter encrustation was assessed visually and by electron microscopy. Bacteriological analysis was performed on the stones and catheter biofilms. P. mirabilis isolates were genotyped by pulsed field gel electrophoresis of restriction enzyme digests of bacterial DNA. RESULTS: Most patients (85%) had been referred because of catheter blockage and in 61 (72%) the catheters were encrusted. P. mirabilis was recovered from 37 of 47 encrusted catheters (79%) that were examined but not from any nonencrusted catheters. Of the 61 patients with encrusted catheters 38 (62%) had bladder stones. Pairs of isolates of P. mirabilis from the stones and the catheter biofilms from 6 patients were genotyped. The DNA profiles of each pair of isolates were identical. CONCLUSIONS: The majority of patients (62%) with recurrent catheter encrustation had bladder stones. The stones harbored the strains of P. mirabilis that rapidly colonize replacement catheters with crystalline biofilm. Flexible cystoscopy to detect and remove stones might help resolve the problem of recurrent catheter encrustation.

Molecular epidemiology of Proteus mirabilis infections of the catheterized urinary tract.

Proteus mirabilis compromises the care of many patients undergoing long-term indwelling bladder catheterization. It forms crystalline bacterial biofilms in catheters which block the flow of urine, causing either incontinence due to leakage or painful distention of the bladder due to urinary retention. If it is not dealt with, catheter blockage can lead to pyelonephritis and septicemia. We have examined the epidemiology of catheter-associated P. mirabilis infections by use of pulsed-field gel electrophoresis (PFGE) of NotI restriction enzyme digests of bacterial DNA. This technique was shown to be more discriminatory than the classical phenotypic Dienes typing technique. We demonstrated that each of 42 isolates from diverse environmental sources and 10 of 12 isolates from blood, wound swabs, and mid-stream urine samples of hospitalized patients had distinct genotypes. Examination of a set of 55 isolates of P. mirabilis, each from a different clinical or environmental source, identified 49 distinct genotypes and 43 Dienes types. The index of discrimination was 0.993 for the PFGE method and 0.988 for the Dienes method. Applying the PFGE method to isolates from catheter-associated urinary tract infections confirmed that the strains present in the crystalline catheter biofilms were identical to those isolated from the same patient's urine. An analysis of samples taken during a prospective study of infections in catheterized nursing home patients revealed that a single genotype of P. mirabilis can persist in the urinary tract despite many changes of catheter, periods of noncatheterization, and antibiotic therapy.

The migration of Proteus mirabilis and other urinary tract pathogens over Foley catheters.

OBJECTIVE: To examine the ability of organisms that infect the catheterized urinary tract to migrate over the surfaces of Foley catheters. MATERIALS AND METHODS: In a simple laboratory model, organisms were challenged to migrate across sections of hydrogel-coated latex, hydrogel/silver-coated latex, silicone-coated latex and all-silicone catheters. The sections (1 cm long) were placed as bridges in channels between blocks of agar and the test organisms inoculated onto the agar adjacent to one side of each bridge. The plates were incubated at 37 degrees C for 24 h and examined for growth of the test organisms on the agar on the other side of the bridges. A collection of swarming, swimming and nonmotile species were tested in the model. The relative mobilities of the test organisms were expressed as migration indices, calculated as the percentage of tests in which bacterial migration was observed over each type of catheter bridge. RESULTS: The swarmer cells of Proteus mirabilis and P. vulgaris migrated successfully (migration indices of 73-100) over all four types of catheter. The migration index of Serratia marcescens swarmers was reduced to 33 over the silver-coated catheters, but these cells crossed over the other catheter surfaces with ease (indices of 100). Pseudomonas aeruginosa was the most mobile of the swimming, non-swarming organisms with indices of 70-22, but this group was less capable of migration than the swarmers. Indices were 0-33 for nonmotile organisms. The mean migration indices for the nine species for each type of catheter were 57 (hydrogel-coated latex), 49 (silver/hydrogel-coated latex), 41 (silicone-coated latex) and 35 (all-silicone). The swarmer cells of P. mirabilis moved through populations of Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus and Enterococcus faecalis, and then migrated over sections of hydrogel-coated latex catheters with little or no reduction in migration index. They were also capable of transporting the nonmotile cells of K. pneumoniae and S. aureus over the catheters. The migration index of P. mirabilis swarmers was substantially reduced in the presence of Ps. aeruginosa and S. marcescens. CONCLUSIONS: Hydrogel coatings facilitate the migration of urinary tract pathogens over catheter surfaces. With the exception of S. marcescens, the incorporation of silver into the hydrogel did not inhibit migration. Swarmer cells were particularly effective at moving over catheters and P. mirabilis swarmers were also capable of transporting other species. This suggests that inhibitors of swarming could be useful in controlling catheter-associated infection and the complications resulting from the spread of bacterial biofilm over catheters.

Studies on the formation of crystalline bacterial biofilms on urethral catheters.

A model of the catheterised bladder was used to test the ability of urease-producing urinary tract pathogens to encrust urethral catheters. Encrustation was assessed by determining the amounts of calcium and magnesium deposited on the catheters and visualised by scanning electron microscopy. Urease-positive Morganella morganii, Klebsiella pneumoniae, and Pseudomonas aeruginosa failed to raise the urinary pH and form crystalline biofilms. In contrast, strains of Proteus mirabilis, Proteus vulgaris, and Providencia rettgeri generated alkaline urine (pH 8.3-8.6) and extensive catheter encrustation within 24 h.