2008年至2010年泌尿系统感染中病原菌的分布及耐药性分析

目的了解泌尿系统感染的病原菌分布及药物耐药性。方法采用法国生物梅里埃公司的VITEK60分析仪对2008年至2010年宁波市妇女儿童医院疑为泌尿系统感染患者的尿液标本进行细菌培养、菌株鉴定,纸片扩散确证试验检测ESBLs。结果 2008年至2010年尿标本中共分离出病原菌1 561株,以大肠埃希菌最多见,占27.2%,其次肺炎克雷伯菌、奇异变形菌、粪肠球菌(D群),各占6.34%、6.28%和6.21%,再次是表皮葡萄球、白色念珠菌和屎肠球菌(D群),各占4.48%、4.36%和3.91%。表皮葡萄球、粪肠球菌(D群)、屎肠球菌(D群)对万古霉素均敏感,对利奈唑烷仅1株粪肠球菌(D群)耐药。3年中,无1例大肠埃希菌对亚胺培南耐药,1株肺炎克雷伯菌和1株奇异变形菌对亚胺培南耐药,其他药物均有不同程度耐药。结论大肠埃希菌是导致泌尿系统感染最常见的致病菌,产ESBLs的菌株已达46.6%。治疗由产ESBLs细菌引起的尿路感染首选亚胺培南和哌拉西林/他唑巴坦;引起尿路感染的革兰阳性菌主要为肠球菌,青霉素可作为治疗粪肠球菌引起的尿路感染,但不适合治疗屎肠球菌引起的尿路感染,耐药率已达95%以上,呋喃妥因、利奈坐烷、万古霉素可作为首选。     

Virulence of a Proteus mirabilis ATF isogenic mutant is not impaired in a mouse model of ascending urinary tract infection

Proteus mirabilis, a common cause of urinary tract infection, produces a number of different fimbriae, including ambient temperature fimbriae (ATF). These fimbriae are optimally expressed at 23 degrees C and their contribution to urinary tract infection has so far remained unknown. In the present study, a clinical isolate of P. mirabilis and an isogenic allelic replacement mutant unable to express ATF were tested for their ability to cause infection in the ascending urinary tract infection model in mice. The atf mutant colonised the urinary tract as well as the wild-type strain and was also able to outcompete the wild-type strain in a co-challenge experiment. Different non-clinical P. mirabilis isolates showed a reactive AtfA band after Western blot analysis using a polyclonal rabbit AtfA antiserum. These data together suggest that ATF does not play a role in P. mirabilis urinary tract infection.     

Molecular mimicry between HLA-DR alleles associated with rheumatoid arthritis and Proteus mirabilis as the Aetiological basis for autoimmunity

Molecular mimicry is one of the pathological mechanisms proposed to explain the association between microorganisms and autoimmune diseases. This review deals with the association between bacteria and rheumatic diseases with a special emphasis on rheumatoid arthritis where upper urinary tract infection by Proteus mirabilis is the possible cause of this severe, arthritic condition. Prospective trials involving anti-Proteus therapy should be carried out.     

Treatment of Infections with Colistimethate Sodium (Coly-Mycin)

Colistimethate sodium (Coly-Mycin) was used in the treatment of 17 patients: 13 had urinary tract infections (two of these had positive blood cultures), three had respiratory tract infections, and one patient had both urinary and respiratory tract infections. In nine of the 17 a foreign body—either a carcinoma, a catheter, or a stone—complicated the infection.The dosage used was 1.1-2.3 mg./lb./day with a maximum in one case of 2.4 g. given over an eight-day period. The organisms so treated included Pseudomonas, six; Aerobacter, six and E. coli, two. Both Pseudomonas and Aerobacter were encountered in three cases.On bacteriological grounds, six patients were cured, eight relapsed, and in three the infecting agent was replaced by another organism. The best responses were obtained in those patients with Pseudomonas infection. Side effects included nausea, vomiting, vertigo, paresthesias, and pain at the site of injection.Colistimethate sodium has a place in the treatment of Gram-negative infections excluding Proteus organisms.     

New aspects of the role of MR/P fimbriae in Proteus mirabilis urinary tract infection

Proteus mirabilis, a common cause of urinary tract infection (UTI), produces a number of different fimbriae including mannose-resistant Proteus-like fimbriae (MR/P). The precise role of different P. mirabilis fimbriae in ascending UTI has not yet been elucidated. In this study, a clinical isolate of P. mirabilis and an isogenic mutant unable to express MR/P were tested using different experimental approaches. They were tested for their ability to cause infection in an ascending co-infection model of UTI and in a haematogenous model in the mouse. In both models, the mutant was less able than the wild-type strain to colonise the lower and upper urinary tracts although infectivity was not abolished. In vitro adherence to uroepithelial cells was also assessed. Significant differences in adherence between both strains were observed at 1 h but not at 15 min post infection. We have also shown that a wild-type strain carries two copies of the mrpA gene. These data reinforce the importance of MR/P fimbriae in P. mirabilis UTI although other virulence factors may be necessary for efficient colonisation and development of infection.     

Site of Action of Nitrofurantoin in Experimental Urinary Tract Infection

The effectiveness of nitrofurantoin in suppressing bacterial growth in the urinary tract was evaluated by using two different experimental models. Pyelonephritis was produced in rats by direct inoculation of 10(4)Escherichia coli in the medulla of left kidney. Ascending urinary tract infection was induced by inoculation into the urinary bladder of 10(7)Proteus mirabilis, after a partial cystectomy. Nitrofurantoin was shown to be effective in suppressing bladder bacteriuria, in preventing ascending pyelonephritis, and also in preventing bacterial multiplication in kidney tissue following direct inoculation.     

Detection of volatile compounds produced by microbial growth in urine by selected ion flow tube mass spectrometry (SIFT-MS)

Selected ion flow tube-mass spectrometry has been used to measure the volatile compounds occurring in the headspace of urine samples inoculated with common urinary tract infection (UTI)-causing microbes Escherichia coli, Proteus vulgaris, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus epidermidis, Klebsiella pneumoniae, Enterococcus faecalis, or Candida albicans. This technique has the potential to offer rapid and simple diagnosis of the causative agent of UTIs.     

Virulence factors in Proteus bacteria from biofilm communities of catheter-associated urinary tract infections

More than 40% of nosocomial infections are those of the urinary tract, most of these occurring in catheterized patients. Bacterial colonization of the urinary tract and catheters results not only in infection, but also various complications, such as blockage of catheters with crystalline deposits of bacterial origin, generation of gravels and pyelonephritis. The diversity of the biofilm microbial community increases with duration of catheter emplacement. One of the most important pathogens in this regard is Proteus mirabilis. The aims of this study were to identify and assess particular virulence factors present in catheter-associated urinary tract infection (CAUTI) isolates, their correlation and linkages: three types of motility (swarming, swimming and twitching), the ability to swarm over urinary catheters, biofilm production in two types of media, urease production and adherence of bacterial cells to various types of urinary tract catheters. We examined 102 CAUTI isolates and 50 isolates taken from stool samples of healthy people. Among the microorganisms isolated from urinary catheters, significant differences were found in biofilm-forming ability and the swarming motility. In comparison with the control group, the microorganisms isolated from urinary catheters showed a wider spectrum of virulence factors. The virulence factors (twitching motility, swimming motility, swarming over various types of catheters and biofilm formation) were also more intensively expressed.     

Combined Staphylococcus-Proteus-Pseudomonas vaccine. II. The toxicity of the vaccine in "chronic" experiments and its ability to protect animals against Proteus and Staphylococcus infections

The combined preparation consisting of the antigenic complexes of staphylococci (1 part), Proteus (1 part) and P. aeruginosa parts) was capable of protecting mice from infection with staphylococci, Proteus and P. aeruginosa and prolonging the survival time of rabbits under the conditions of the development of staphylococcal sepsis. The staphylococcal component of the combined preparation possessed adjuvant activity, increasing the immunogenicity of Proteus antigen. The combined vaccine enhanced a short-time increase in the nonspecific resistance of the animals. The moderate toxicity of the preparation permits making multiple injections of the preparation to mice without inhibiting the weight gain of the animals.     

Surface immobilization of kanamycin-chitosan nanoparticles on polyurethane ureteral stents to prevent bacterial adhesion

Bacterial adhesion is a major problem that can lead to the infection of implanted urological stents. In this study, kanamycin-chitosan nanoparticles (KMCSNPs) were immobilized on the surface of a polyurethane ureteral stent (PUS) to prevent urinary bacterial infection. KMCSNPs were synthesized using the ionic gelation method. The synthesized KMCSNPs appeared spherical with a ζ-average particle size of 225 nm. KMCSNPs were immobilized on the PUS surface by covalent immobilization techniques. The surface-modified PUS was characterized using attenuated total reflectance Fourier transform infrared spectroscopy, field emission scanning electron microscopy, and energy dispersive X-ray spectroscopy. The surface-modified PUS showed significantly increased antibacterial activity against Escherichia coli MTCC 729 and Proteus mirabilis MTCC 425 relative to the surface of an unmodified PUS. These findings suggest that the KMCSNP-immobilized PUS has the potential to prevent bacterial infection in the human urinary tract.     

Preventive and therapeutic administration of an indigenous <Emphasis Type="Italic">Lactobacillus</Emphasis> sp. strain against <Emphasis Type="Italic">Proteus mirabilis</Emphasis> ascending urinary tract infection in a mouse model

Probiotics are increasingly being considered as non-pharmaceutical and safe potential alternatives for the treatment and prevention of a variety of pathologies including urinary tract infections. These are the most common infections in medical practice and are frequently treated with antibiotics, which have generated an intense selective pressure over bacterial populations. Proteus mirabilis is a common cause of urinary tract infections in catheterised patients and people with abnormalities of the urinary tract. In this work we isolated, identified and characterised an indigenous Lactobacillus murinus strain (LbO2) from the vaginal tract of a female mouse. In vitro characterisation of LbO2 included acid and bile salts tolerance, growth in urine, adherence to uroepithelial cells and in vitro antimicrobial activity. The selected strain showed interesting properties, suitable for its use as a probiotic. The ability of LbO2 to prevent and even treat ascending P. mirabilis urinary tract infection was assessed using an experimental model in the mouse. Kidney and bladder P. mirabilis counts were significantly lower in mice preventively treated with the probiotic than in non-treated mice. When LbO2 was used for therapeutic treatment, bladder counts of treated mice were significantly lower although no significant differences were detected in P. mirabilis kidney colonisation of treated and non-treated animals. These results are encouraging and prompt further research related to probiotic strains and the basis of their effects for their use in human and animal health.     

Trimethoprim resistance in Finland after five years' use of plain trimethoprim.

A total of 1388 urinary bacterial pathogens were tested for resistance to plain trimethoprim after five years'' use of this drug for prophylaxis against urinary tract infections. Samples were obtained in Turku, Finland, where use of the drug is much greater than in other parts of Finland. Resistance to trimethoprim (greater than 8 mg/l; agar-dilution method) occurred in 20.3% of strains isolated from outpatients and 39.8% of strains isolated from inpatients. Escherichia coli and Micrococcus showed low incidences of resistance (11% and 13% respectively in ouptatients and 23% and 19% respectively in inpatients); Enterobacter, Streptococcus faecalis, and Staphylococcus epidermidis occupied an intermediate position; and Proteus mirabilis and Klebsiella were resistant in 41-76% of cases. Similar incidences of resistance were observed to sulphamethoxazole-trimethoprim, sulphamethoxazole, ampicillin, and nitrofurantoin. These findings together with the rare occurrence of side effects and convenient dosage confirm the usefulness of plain trimethoprim for urinary tract infection.     

Mouse models of short- and long-term foreign body in the urinary bladder: analogies to the bladder segment of urinary catheters.

Catheter-associated bacteriuria is the most common infection occurring in hospitals, where urethral catheters are generally in place for a few days, and in nursing homes, where catheters may be in place for months or years. We developed murine models with intrabladder urinary catheters for studying complications of bacteriuria in short- and long-term catheterization. In the short-term model, a catheter segment was inserted transurethrally and lay free within the bladder lumen. Half of the animals expelled segments during a 2-to-7-day period, durations similar to catheterizations in hospitalized patients. For studies of long-term catheter use, the catheter segment was secured within the bladder by a single suture for up to 12 months. Antibiotics administered for 7 days after catheter placement and housing mice in cages with wire screen floors reduced spontaneous bacteriuria to an acceptably low incidence rate of only 7%. Proteus mirabilis bacteriuria of high concentration provoked the same complications that are common in patients with long-term catheters: acute pyelonephritis, chronic renal inflammation, and struvite stone formation. These models allow inoculation of the bacteria of interest and are suitable for studies of short- and long-term foreign body-associated bacteriuria and its complications.     

Clinical and economic expedience of ertapenem therapy of complicated urinary tract infection

Clinical and economic investigation of various antibiotics use in the treatment of complicated urinary tract infection (CUTI) was performed under the Russian economic environment. The drugs of comparison were ertapenem, ceftriaxone and levofloxacin. Direct costs and their structure were shown, and the cost efficiency was calculated. Alternative analysis and one-side susceptibility analysis were performed. In complicated urinary tract infections when the major pathogens were Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis it was clinically and economically reasonable to start the treatment with ceftriaxone or ertapenem, while levofloxacin could be an alternative strategy. When the effects of the acquired resistance on the treatment effectiveness were evaluated (SIS model) it was shown that the pathogens susceptibility to ertapenem was preserved for a significantly longer time than that to ceftriaxone or levofloxacin (60 months). Such a parameter may serve as an additional evidence of the reasonable use of ertapenem as the starting treatment of CUTI.     

Síndrome de la bolsa de orina púrpura en dos pacientes institucionalizados

Purple urine bag syndrome (PUBS) is an uncommon but particularly striking phenomenon characterised by a chemical reaction involving the urine, plastic and certain enzymes from some sulphatase- and phosphatase-producing bacteria, including Proteus mirabilis, Escherichia coli and Morganella morganii, amongst others. Following this reaction, the catheter and the bag may be stained red, blue or purple. This phenomenon tends to occur in patients with multiple pathology and with urinary catheters, as part of a urinary tract infection. We describe two clinical cases of PUBS in institutionalised patients with permanent urinary catheters.     

ZapA, the IgA-degrading metalloprotease of Proteus mirabilis, is a virulence factor expressed specifically in swarmer cells

The IgA-degrading metalloprotease, ZapA, of the urinary tract pathogen Proteus mirabilis is co-ordinately expressed along with other proteins and virulence factors during swarmer cell differentiation. In this communication, we have used zapA to monitor IgA protease expression during the differentiation of vegetative swimmer cells to fully differentiated swarmer cells. Northern blot analysis of wild-type cells and beta-galactosidase measurements using a zapA:lacZ fusion strain indicate that zapA is fully expressed only in differentiated swarmer cells. Moreover, the expression of zapA on nutrient agar medium is co-ordinately regulated in concert with the cycles of cellular differentiation, swarm migration and consolidation that produce the bull's-eye colonies typically associated with P. mirabilis. ZapA activity is not required for swarmer cell differentiation or swarming behaviour, as ZapA- strains produce wild-type colony patterns. ZapA- strains fail to degrade IgA and show decreased survival compared with the wild-type cells during infection in a mouse model of ascending urinary tract infection (UTI). These data underscore the importance of the P. mirabilis IgA-degrading metalloprotease in UTI. Analysis of the nucleotide sequences adjacent to zapA reveals four additional genes, zapE, zapB, zapC and zapD, which appear to possess functions required for ZapA activity and IgA proteolysis. Based on homology to other known proteins, these genes encode a second metalloprotease, ZapE, as well as a ZapA-specific ABC transporter system (ZapB, ZapC and ZapD). A model describing the function and interaction of each of these five proteins in the degradation of host IgA during UTI is presented.     

Immunogenicity of a vaccine made from soluble Proteus antigens

Hemagglutinins with different specificity were determined in 270 subjects: of these, 101 were examined in the time course before and after immunization with Proteus vaccine prepared from soluble antigens. The preparation possessed pronounced immunological potency and stimulated the formation of antibodies to the vaccine strain, to heterologous Proteus strains and to the common antigens of Gram-negative bacteria. The combined scheme of the subcutaneous and local administration of the above-mentioned Proteus vaccine ensured an increase in the synthesis of IgM and IgA in patients with Proteus wound infection. Active immunization ensured an essential rise in the level of hemagglutinins to Re-glycolipid in donors and oncological patients not infected with Proteus, but did not ensure the statistically significant shifts in the antibody level to Re-glycolipids in patients with chronic Proteus infection.     

Prevalence of multidrug-resistant Proteus spp. strains in clinical specimens and their susceptibility to antibiotics

Proteus sp. are opportunistic microorganisms which cause urinary tract and wounds infections, bacteriaemia and sepsis. The aim of this study was analysis of prevalence of multidrug-resistant Proteus sp. strains in clinical specimens and evaluation of their susceptibility to selected antibiotics. The study was carried out of 1499 Proteus sp. strains were isolated in 2000-2003 from patients of departments and dispensaries of the University Hospital CM in Bydgoszcz UMK in Torun. The strains were identified on the basis of appearance of bacterial colonies on bloody and McConkey's agars, movement ability, indole and urease production and in questionable cases biochemical profile in ID GN or ID E (bio-Mérieux) tests was also included. Antibiotic susceptibility was tested by disk diffusion method. Isolated strains were regarded as multidrug-resistant when they were resistant to three kinds of antibiotics at least. Received Proteus sp. the most frequently belonged to P. mirabilis species (92.3%). Most of these bacteria were isolated from urine from patients of Rehabilitation Clinic. All of multidrug-resistant strains were resistant to penicillins and cephalosporins, 98.9% to co-trimoxazole, 77.7% to quinolones, 63.8% to tetracyclines, 38.5% to aminoglycosides, 19.3% to monobactams and 3.4% to carbapenems. Almost 25% multidrug-resistant Proteus sp. produced ESBL.     

Parathyphoid B-antigen containing a complex of protective surface antigens]

The authors obtained a complex antigen from paratyphoid B bacilli containing complete O-, K- and H-antigens. The preparation was nontoxic and was characterised by marked antigenic properties. In intravenous and oral administration it stimulated production of specific O-, K-, and H-antibodies in high titres. Complex paratyphoid B antigen possessed a marked protective activity and provided intense immunity in subcutaneous and oral administration to experimental animals.     

Characteristics of bacteria in the genus Proteus isolated from patients with sporadic and group intestinal diseases

The biochemical and biological properties of 148 Proteus strains isolated from patients both in sporadic intestinal infections and in a case of group infection in children's hospital was studied. The study revealed that the etiological factor of the group infection was P. mirabilis belonging to rare serovar 48:2. Proteus organisms isolated in sporadic infections belonged to a great number of serovars. No relationship between the isolated serovar and the nosological form of the intestinal disease was established. Among the Proteus strains under study, 82 strains showed atypical biochemical properties in 1 test or more. No correlation between the clinical diagnosis and the occurrence of atypical strains was established.