Genotypic and phenotypic profiles of virulence factors and antimicrobial resistance of Proteus mirabilis isolated from chicken carcasses: potential zoonotic risk.

Proteus mirabilis is an opportunistic pathogen often associated with a variety of human infections acquired both in the community and in hospitals. In this context, the present work aimed to evaluate the genotypic and phenotypic characteristics of the virulence factors and antimicrobial resistance determinants of 32 P. mirabilis strains isolated from chicken carcasses in a poultry slaughterhouse in the north of the state of Paraná, Brazil, in order to assess a potential zoonotic risk. The isolates presented a variety of virulence genes that contribute to the development of infection in humans. The mrpA, pmfA, atfA (fimbriae), ireA (siderophores receptor), zapA, ptA (Proteases), and hpmA (hemolysin) genes were found in 32 (100%) isolates and ucaA (fimbriae) in 16 (50%). All isolates showed aggregative adherence in HEp-2 cells and formed biofilms. Of all strains, 27 (84.38%) showed cytotoxic effects in Vero cells. Antimicrobial susceptibility was tested using 20 antimicrobials, in which 25 (78.13%) strains were considered multidrug-resistant. The presence of blaESBL and blaampC genes conferring resistance to ß-lactams and qnr to quinolones were also detected in the isolates after presumption in the phenotypic test, in which 7 (21.88%) isolates contained the CTX-M-2 group, 11 (34.38%) contained CIT group and 19 (59.38%) contained qnrD. Therefore, chicken carcasses contaminated with P. mirabilis may pose a health risk to the consumer, as these isolates have a variety of virulence and antimicrobial resistance characteristics that can be found in P. mirabilis strains isolated from human infections.

Dissemination of Multidrug-Resistant Proteus mirabilis Clones Carrying a Novel Integron-Borne blaIMP-1 in a Tertiary Hospital.

This study aimed to characterize multidrug-resistant Proteus mirabilis clones carrying a novel class 1 integron-borne blaIMP-1 In1359 was inserted into a large conjugative plasmid that also carried blaCTX-M-2 The production of carbapenemases in Enterobacteriaceae that are intrinsically resistant to polymyxins and tigecycline is very worrisome, representing a serious challenge to clinicians and infection control teams.

Regional Spread of CTX-M-2-Producing Proteus mirabilis with the Identical Genetic Structure in Japan.

AIMS: In this study, we analyzed the molecular epidemiology of extended-spectrum ß-lactamase (ESBL)-producing Proteus mirabilis isolates collected from the central region of Japan. MATERIALS AND METHODS: Between 2005 and 2012, 820 clinical P. mirabilis isolates were obtained from ten acute care hospitals in Japan. We characterized ESBL confirmatory test-positive isolates by sequencing the ESBL genes and their flanking regions, detecting plasmid replicons, and performing pulsed-field gel electrophoresis (PFGE). RESULTS: Ninety-six isolates (12%) were positive according to the ESBL confirmatory test; all these isolates possessed blaCTX-M-2 with the same flanking structure of upstream ΔISEcp1 and a downstream region identical to downstream blaKLUA-1. IncT was the prevalent, and only, replicon found in 63 isolates. PFGE analysis detected eight clusters with more than one isolate, among which three included 56 isolates and six included isolates from multiple hospitals. CONCLUSION: CTX-M-2-producing P. mirabilis with an identical genetic structure flanking blaCTX-M-2 is dominant in this Japanese region, and there is evidence for the clonal spread of isolates.

Molecular Epidemiology and Genetic Characteristics of Various blaPER Genes in Shanghai, China.

We describe the genetic characteristics and possible transmission mechanism of blaPER in 25 clinical Gram-negative bacilli in Shanghai. blaPER, including blaPER-1, blaPER-3, and blaPER-4, was located chromosomally or in different plasmids. Tn1213 harboring blaPER-1 was first identified in two Proteus mirabilis isolates in China. The other blaPER variants were preceded by an ISCR1 element inside the complex class 1 integron associated with IS26, Tn21, Tn1696, and a miniature inverted-repeat transposable element.

Antibiotic resistance pattern among biofilm producing and non producing Proteus strains isolated from hospitalized patients; matter of hospital hygiene and antimicrobial stewardship.

A retrospective study on antimicrobial susceptibility and biofilm production were carried out for eighty eight strains of Proteus strains isolated from UTI and other hospital samples during April 2011-April 2012. The antibiotic susceptibility was carried out by Kirby-Bauer disk diffusion and MIC by E-test. Biofilm production was measured by microtiter method and confirmed by Scanning electron microscopy. Plasmids from biofilm producing isolates were detected by alkaline lysis technique. From 88 patients infected by proteus species, 58% were female and 42% were mail. The most frequent age range was 20-29 (77.39%) and the least were 60-69 years old (3.4%) (p = 0.05). Eighty one isolates were identified as P. mirabilis while, 7 identified as P. vulgaris. 67.04% [n = 59] of the isolates showed MIC range (16-32 +/- 0.05 microg mL(-1)) to ceftriaxone, 46.59% [n = 41] exhibited least MIC range to chloramphenicol (8-64 +/- 0.08 microg mL(-1)). 31% [n = 28] of the isolates also exhibited MIC range 1-4 microg mL(-1) to ciprofloxacin. 17% [n = 15] of the isolates exhibited strong biofilm while, 6% [n = 6] did not show any biofilm (p < or = 0.05). Plasmid isolation from biofilm producing isolates revealed that stains number 19, 24 and 87' that produced strong biofilm carried similar high M. Wt. plasmid. From above results it can be concluded that the majority of Proteus isolated from UTI patients were belong to P. mirabilis. Ciprofloxacin was the most effective antibiotic for treatment of the infected patients. Limited number of the isolates could produce strong biofilm that were bearing plasmids. Majority of the biofilm producing isolates were also resistance at least to 4 antibiotics routinely prescribed in our hospital.

Rapidly spreading CTX-M-type beta-lactamase-producing Proteus mirabilis in Japan.

In recent years, increased isolation of extended-spectrum beta-lactamase (ESBL)-producing Proteus mirabilis has been reported in Japan. We undertook an investigation to determine the prevalence of ESBL-producing P. mirabilis isolated in Japan and to characterise the genotype. Seventy-four P. mirabilis isolates recovered from specimens at 54 hospitals in Japan between March and October 2006 were included in the study. Of the 74 P. mirabilis isolates examined, 28 (37.8%) were ESBL-producers. The bla(CTX-M-2) gene was found in 27 isolates, whilst 1 isolate possessed bla(CTX-M-3). Amongst the 28 ESBL-producers, 25 (89.3%) were non-susceptible to ciprofloxacin, whilst 11 (23.9%) of 46 ESBL-non-producing isolates were non-susceptible to ciprofloxacin. Pulsed-field gel electrophoresis (PFGE) analysis of the 28 ESBL-producing isolates from 19 hospitals revealed 17 clusters. The same PFGE type was observed in two or more hospitals especially in the greater Tokyo area, suggesting possible clonal spread and the need for monitoring to determine whether emergence of a dominant clone occurs. Our results show that in Japan there is a high prevalence of CTX-M-type beta-lactamase-producing P. mirabilis. Moreover, these isolates are characterised by reduced susceptibility to fluoroquinolones.

Identification of plasmid-encoded extended spectrum beta-lactamases produced by a clinical strain of Proteus mirabilis.

Emergence and dissemination of multiresistant strain of Proteus mirabilis have made infections treatment more difficult that this bacterium is responsible. The aim of this study is to determine the implication of the enzymatic mechanism and to describe the properties of ESBLs (extended spectrum beta-lactamases). A clinical strain of Proteus mirabilis SM514 isolated in the intensive care unit at the Military hospital in Tunisia during the period 2004 was found to be highly resistant to cephalosporins and penicilins. Cells sonicate of the isolate hydrolysed cefotaxime more efficiently than ceftriaxone and ceftazidime and had three beta-lactamases bands of approximate of isoelectric points (pI) of 5.4; 5.6 and superior to 7.6. The specific activities (AS) vary from 5.26 to 7.77U/mg of protein respectively for cefotaxime and the benzylpenicillin. These activities are inhibited by the clavulanic acid and the sulbactam. The values of the IC(50) are respectively 3.7 and 11.7muM. Only the beta-lactamases of pI 5.4 and superior to 7.6 hydrolyze the cefotaxime. Transformant produces the ESBLs of pI 5.4; 7.45 and greater than 7.6. The genes coding for this enzymes are carried by a transferable plasmids.

Transmission in the community of clonal Proteus mirabilis carrying VIM-1 metallo-beta-lactamase.

OBJECTIVES: Several infections among patients attending our outpatient clinic were caused by imipenem-resistant Proteus mirabilis that were phenotypically metallo-beta-lactamase (MBL)-positive. The aim of the study was to investigate this extrahospital dissemination and the mechanisms of resistance to carbapenems. METHODS: During a 1 year period (December 2005-December 2006), the characteristics of the outpatients with infections caused by isolates of P. mirabilis with reduced susceptibility to imipenem (MIC > 4 mg/L) were prospectively collected. The isolates were tested by agar dilution MICs, phenotypic MBL testing and enterobacterial repetitive intergenic consensus PCR. PCR assays and nucleotide sequencing were used for the identification of bla gene types and mapping of the integron carrying the MBL gene. The location of the MBL allele was investigated by mating experiments, plasmid analysis and hybridization of the Southern-blotted plasmid extract with a bla(VIM-1) probe. RESULTS: During the study, 12 MBL-positive P. mirabilis isolates were recovered from urinary tract infections of community patients. In all cases, the patients had a previous hospitalization in a Greek regional hospital and had received fluoroquinolones and/or aminoglycosides and beta-lactams. MICs of imipenem ranged from 32 to >128 mg/L, whereas those of meropenem ranged from 1 to 8 mg/L and those of ertapenem ranged from 0.5 to 4 mg/L. The isolates originated from the same clonal strain and harboured a bla(VIM-1) gene in a common integron structure. Conjugation experiments, plasmid analysis and hybridization assays indicated the chromosomal location of the bla(VIM-1) gene. CONCLUSIONS: This is the first study that documents transmission in the extrahospital setting of acquired MBL-producing Gram-negatives causing community-onset infections.

Nosocomial outbreak of infections by Proteus mirabilis that produces extended-spectrum CTX-M-2 type beta-lactamase.

Nineteen multidrug-resistant Proteus mirabilis strains were isolated from 19 patients suffering from infections probably caused by P. mirabilis. These strains were recovered from urine or other urogenital specimens of 16 inpatients and three outpatients with a hospitalization history in a urology ward of Funabashi Medical Center, from July 2001 to August 2002. These strains demonstrated resistance to cefotaxime, ceftriaxone, cefpodoxime, and aztreonam, while they were highly susceptible to ceftazidime (MIC,

Post-keratoplasty endophthalmitis caused by Proteus mirabilis.

A diabetic patient who underwent a triple procedure (penetrating keratoplasty, cataract extraction and posterior chamber intraocular lens implantation), developed endophthalmitis caused by Proteus mirabilis. The source of infection was the infected donor cornea, which was imported from Sri Lanka. The organism was resistant to gentamicin, which was the only antibiotic present in the storage medium. On top of an aggressive antibiotic treatment regime (topical, systemic and intravitreal), exchange of the infected with a fresh corneal graft and pars plana vitrectomy were performed. The patient had a speedy recovery and the visual outcome was 20/40 two and a half years after the incident. To our knowledge, post-keratoplasty endophthalmitis caused by P. mirabilis has never been reported in the literature. We report herein such a case with good visual outcome. Newer storage medium, such as Optisol GS, might have a role in preventing keratoplasty-related infection.

Proteus penneri urosepsis in a patient with diabetes mellitus.

Proteus penneri has been isolated from many different clinical sources, including surgical wound infections, urine, and blood. We describe the first reported case of P. penneri nosocomial urosepsis in a patient with diabetes. P. penneri was subsequently isolated from bronchoalveolar lavage fluid and a pulmonary artery catheter tip.

Retrograde transmission of Proteus mirabilis during platelet transfusion and the use of arbitrarily primed polymerase chain reaction for bacteria typing in suspected cases of transfusion transmission of infection.

BACKGROUND: When bacteria are found, after a platelet transfusion, in the recipient's blood as well as in the platelet concentrate (PC), a causal relationship is normally suspected, with the PC as the causative agent. The other alternative, that the patient has bacteremia and contaminated the PC, is less well documented in the literature. CASE REPORT: Arbitrarily primed polymerase chain reaction (AP-PCR) was used for testing strains of Proteus mirabilis isolated from a patient's blood before and after a platelet transfusion and from the PC. Because of a febrile reaction after a platelet transfusion, bacterial culture was performed on the PC used, showing growth of P. mirabilis. The same species was found in the patient's blood after the transfusion. Posttransfusion sepsis caused by a contaminated PC was suspected, and anti-sepsis treatment was given to the recipient. Later, it became apparent that the patient had had bacteremia before the transfusion and that P. mirabilis was one of the species in the isolate. With AP-PCR, the identity of the three P. mirabilis isolates could be distinguished. CONCLUSION: AP-PCR is a useful technique for distinguishing the identity of bacterial isolates from patients and blood components. A patient with bacteremia can contaminate a PC in conjunction with a platelet transfusion. With AP-PCR, the PC could be ruled out as the cause of the posttransfusion sepsis.

Development of a catheterised bladder model to evaluate urinary drainage equipment.

A catheterised bladder model was developed to examine the design features of urinary drainage bags. The drainage bag of 4 commonly used products was inoculated with either Escherichia coli or Proteus mirabilis. The speed of retrograde contamination towards the "bladder" was determined over 96 h. The effect of lifting the drainage bag above the level of the "bladder" was examined also. Within 4 days micro-organisms were detected in the backflow valves/drip chambers of all products, but contamination had not progressed as far as the catheter. Ps. mirabilis appeared to migrate more rapidly than Esch. coli. Lifting the drainage bag had no effect on the migration of either organism. The implications of these results for clinical practice are discussed.

Arbitrarily primed polymerase chain reaction provides rapid differentiation of Proteus mirabilis isolates from a pediatric hospital.

During a systematic survey, maternal carriage of Proteus mirabilis was found over a 25-day period in 18 pregnant women admitted to the delivery ward of our hospital maternity. Five neonates born to these mothers were found to be colonized with P. mirabilis. We report here on the use of DNA fingerprinting by the arbitrarily primed polymerase chain reaction technique (AP-PCR) for the epidemiological investigation of this sudden outbreak. This approach was compared with the analysis of restriction fragment length polymorphisms of ribosomal DNA regions (ribotyping). Results of the AP-PCR and of ribotyping were in complete agreement in showing the genetic unrelatedness of the isolates obtained from each mother. Moreover, the results showed mother-to-infant vertical transmission of P. mirabilis in the neonates. AP-PCR is a rapid and discriminative method which seems particularly well suited to the epidemiological study of P. mirabilis.

The Dienes effect as an epidemiological tool in a paraplegic unit.

The authors study the Dienes effect in 136 P.mirabilis and 2 P.vulgaris strains, isolated from different infection or colonization sites, in 27 spinal cord injured patients, admitted to a Paraplegic Unit, during a 9 month period. It is shown that 7 cross-infection and 18 cross-contamination episodes occurred, affecting 13 of 27 patients, which indicates the great spreading capacity of these microorganisms; 97% of the results obtained were observed once again after 6 months' storage of strains. In addition, in 7 of these strains their resistance to several antibiotics was cured by treating them with acriflavine salts, demonstrating that the Dienes effect persists even when resistance to antibiotics is modified; the authors conclude that the Dienes effect is an accurate stable epidemiological tool to identify cross-infections and its origins, and to facilitate the interruption of the chain of infection.

Effects of environment on infection in burn patients.

Burn patients in an early cohort (n = 173) treated in an intensive care ward without separate enclosures were compared with a later cohort (n = 213) treated in a renovated unit with separate bed enclosures. The number of patients developing infection was significantly reduced in the late group. Observed mortality was compared with mortality predicted on the basis of burn size and age alone. Reduction in observed compared with predicted mortality, inapparent in the early group, was seen in the late group and was restricted to the subgroup of patients with predicted mortality of 25% to 75%, in which the observed mortality of 28.3% was less than the predicted mortality of 48.7%. The incidence of infected patients was reduced from 58.1% in the early cohort to 30.4% in the late cohort. In comparison of the early cohort with the late cohort, the overall proportion of patients with bacteremia was reduced from 20.1% to 9.4%, while the incidences of both pneumonia and burn wound invasion remained unchanged. Providencia and Pseudomonas species, endemic in the early cohort, were eliminated in the late cohort. Reduction of infection by environmental manipulation in burn patients was possible and was associated with improved survival.

The use of the Dienes test in the epidemiology of proteus urinary tract infections in patients with spinal cord disease.

The incidence of urinary tract infections in 47 patients with spinal cord injuries (11 women and 36 men) was studied and the aetiological agents identified. A high incidence of Proteus mirabilis infection was found and both auto- and cross-infection were identified by the Dienes test.