Rapid identification and antimicrobial susceptibility profiling of Gram-positive cocci in blood cultures with the Vitek 2 system

Rapid identification and antimicrobial susceptibility profiling of the bacteria in blood cultures can result in clinical and financial benefits. Addition of saponin to the fluid from blood culture bottles promotes the recovery of the bacteria and thus may shorten the turnaround time of the microbiological analyses. In this study we compared the identification and susceptibility profiles of saponin-treated and untreated (standard method) blood cultures monomicrobial for Gram-positive cocci using Vitek 2. We concordantly identified 49 (89%) of 55 monobacterial cultures using the results with the standard method as reference. Complete categorical agreement between the susceptibility profiles with the new and the standard method was found for 26 (53%) of 49 isolates, while discrepancies were seen for 23 (47%) cultures. E-tests indicated that the new method resulted in a correct susceptibility profile for 8 (35%) of these 23 blood cultures. Therefore, 34 (69%) of 49 cultures showed a concordant/correct susceptibility profile for all antimicrobials with an overall error rate of 2.3%. Thus, addition of saponin to the fluid from blood culture bottles of the Bactec 9240 leads to the rapid (results available ≥12 hours earlier) and reliable identification and susceptibility profiling of Gram-positive cocci in blood cultures with Vitek 2

Serratia marcescens internalization and replication in human bladder epithelial cells

BACKGROUND: Serratia marcescens, a frequent agent of catheterization-associated bacteriuria, strongly adheres to human bladder epithelial cells in culture. The epithelium normally provides a barrier between lumal organisms and the interstitium; the tight adhesion of bacteria to the epithelial cells can lead to internalization and subsequent lysis. However, internalisation was not shown yet for S. marcescens strains. METHODS: Elektronmicroscopy and the common gentamycin protection assay was used to assess intracellular bacteria. Via site directed mutagenesis, an hemolytic negative isogenic Serratia strain was generated to point out the importance of hemolysin production. RESULTS: We identified an important bacterial factor mediating the internalization of S. marcescens, and lysis of epithelial cells, as the secreted cytolysin ShlA. Microtubule filaments and actin filaments were shown to be involved in internalization. However, cytolysis of eukaryotic cells by ShlA was an interfering factor, and therefore hemolytic-negative mutants were used in subsequent experiments. Isogenic hemolysin-negative mutant strains were still adhesive, but were no longer cytotoxic, did not disrupt the cell culture monolayer, and were no longer internalized by HEp-2 and RT112 bladder epithelial cells under the conditions used for the wild-type strain. After wild-type S. marcescens became intracellular, the infected epithelial cells were lysed by extended vacuolation induced by ShlA. In late stages of vacuolation, highly motile S. marcescens cells were observed in the vacuoles. S. marcescens was also able to replicate in cultured HEp-2 cells, and replication was not dependent on hemolysin production. CONCLUSION: The results reported here showed that the pore-forming toxin ShlA triggers microtubule-dependent invasion and is the main factor inducing lysis of the epithelial cells to release the bacteria, and therefore plays a major role in the development of S. marcescens infections

Antimicrobial susceptibilities of 40 isolates of Bacillus anthracis isolated in Turkey

Forty clinical isolates of Bacillus anthracis were studied. The MIC90 values of penicillin G, doxycycline, ciprofloxacin, gatifloxacin, and levofloxacin were 0.016, 0.03, 0.06, 0.06 and 0.12 mg/l, respectively. Susceptibilities suggest that the quinolones may also be considered as an alternative therapy for anthrax. (C) 2003 Elsevier Science B.V. and the International Society of Chemotherapy. All rights reserved

Prospective evaluation of blood cultures in a Turkish university hospital: epidemiology, microbiology and patient outcome

The aims of this prospective study were to: (1) determine the rate of blood culture contamination; (2) describe and compare the epidemiologic, clinical and microbiological characteristics of hospital- and community-acquired bloodstream infections; and (3) determine the mortality resulting from bloodstream infections. The rate of true bacteremia was 12.1%, and 10.7% of cultures were contaminated. Of the 567 episodes of bloodstream infection, 73.4% were hospital-acquired, and 26.6% were community-acquired. The most commonly isolated microorganisms were staphylococci (44%, methicillin resistant 69.4%), enterococci (15%) and Escherichia coli (12.5%) in hospital-acquired episodes, and Brucella spp. (21.9%), E. coli (19.2%) and Staphylococcus aureus (14.6%, methicillin resistant 9.1%) in community-acquired episodes. While the overall mortality rate was 25.4%, death attributable to bloodstream infections was 16.6% in hospital-acquired episodes and 13.9% in community-acquired episodes. The highest mortality occurred in patients with bacteremia due to Pseudomonas aeruginosa (37.5%) in hospital-acquired episodes, and in patients with bacteremia due to Streptococcus pneumoniae (50%) in community-acquired episodes. Underlying diseases, severity of illness, presence of bladder catheter, previous use of antibiotics, tracheal intubation and adequacy of treatment were found to be significantly associated with death

Fatal peritonitis due to Trichoderma sp in a patient undergoing continuous ambulatory peritoneal dialysis

We describe a rare case of peritonitis caused by an unusual fungus, Trichoderma sp., in a patient on continuous ambulatory peritoneal dialysis. Management of the patient consisted of Tenckhoff catheter removal and antifungal chemotherapy, but the patient died

Evaluation of susceptibility patterns and BRO beta-lactamase types among clinical isolates of Moraxella catarrhalis

The aims of this study were to detect BRO beta-lactamase types and to evaluate any correlation with the susceptibility patterns of 90 clinical isolates of Moraxella catarrhalis. The overall prevalences of the bro-1 and bro-2 genes were 78% and 12%, respectively. Penicillin G MICs for BRO-1(+) isolates were significantly higher than those for BRO-2(+) isolates. All the isolates were susceptible to amoxycillin-clavulanate, levofloxacin and cefixime. Resistance to clarithromycin, tetracycline and trimethoprim-sulphamethoxazole was 1.1%, 2.2% and 1.1%, respectively. One-step, length-based PCR was an efficient method to screen for BRO beta-lactamase genes

The efficacy of intravitreal piperacillin/tazobactam in rabbits with experimental Staphylococcus epidermidis endophthalmitis: A comparison with vancomycin

Background: To investigate the efficacy of intravitreal piperacillin/tazobactam in rabbit eyes with experimental S. epidermidis endophthalmitis and to compare the outcomes with intravitreal vancomycin application. Material and Methods: Twenty-four New Zealand white albino rabbits were divided into three equal groups (n=8 in each), and the right eyes received 0.1-ml intravitreal injections of S. epidermidis suspension. The left eyes served as uninfected controls and were injected with 0.1 ml of saline solution. The right eyes of rabbits in group 1 were treated with intravitreal injection of 250 mu g/0.1 ml piperacillin/tazobactam 24 h after intravitreal inoculation of S. epidermidis whereas group 2 eyes received intravitreal 1 mg/0.1 ml vancomycin. Group 3 eyes received no treatment and served as infected controls. Clinical examination of the eyes in each group was performed on the 1st, 3rd and 6th day after the inoculation of S. epidermidis. On the 6th day, 0.1-ml vitreous aspirates were obtained for microbiological analysis, and then the eyes were enucleated for histopathological evaluation. Results: There were no statistically significant differences in mean clinical scores between the groups on the first day after S. epidermidis inoculation (p>0.05). On the 6th day, the mean clinical score of group 3 was significantly higher ( p0.05). Conclusion: Intravitreal application of 250 mu g/0.1 ml piperacillin/tazobactam seems to be approximately equally effective with intravitreal 1 mg/0.1 vancomycin application in the treatment of experimental S. epidermidis endophthalmitis. Therefore, intravitreal piperacillin/tazobactam may be an alternative therapeutic option in the treatment of S. epidermidis endophthalmitis. Copyright (C) 2005 S. Karger AG, Basel

A comparison of intravitreal piperacillin/tazobactam with ceftazidime in experimental Pseudomonas aeruginosa endophthalmitis

In the present study, we aimed at comparing the efficacies of intravitreal piperacillin/tazobactam and ceftazidime applications in the treatment of experimental Pseudomonas aeruginosa endophthalmitis in rabbit eyes. Twenty-four New Zealand white albino rabbits were divided into three groups (n=8 in each), and the right eyes received 0.1 ml intravitreal injections of P. aeruginosa suspension. The left eyes served as uninfected control and were injected with 0.1 ml of saline solution. The right eyes of rabbits in group 1 were treated with intravitreal injection of 250 mu g/0.1 ml piperacillin/tazobactam 24 hr after intravitreal inoculation of P. aeruginosa, whereas group 2 eyes received intravitreal 1 mg/O (.) 1 ml ceftazidime. Group 3 eyes received no treatment and served as infected controls. Clinical, microbiological and histopathological evaluations of the eyes in each group were performed on the 1st, 3rd, and 6th day after the inoculation of P. aeruginosa. The mean clinical scores of each group were similar at the first day after P. aeruginosa inoculation (P > 0.05). At the 6th day, there was no statistically significant difference in mean clinical scores between group 1 and 2, but mean clinical score of group 3 was significantly higher (P 0.05). Group 3 eyes had a significantly more CFU/ml and higher histopathological score (for each, P < 0.001). In conclusion, intravitreal application of 250 mu g/0.1 ml piperacillin/tazobactam seems to be effective in the treatment of P. aeruginosa endophthalmitis in rabbits, but is not superior to intravitreal ceftazidime application. Therefore, intravitreal piperacillin/tazobactam may be a useful alternative to ceftazidime for pseudomonal endophthalmitis. (c) 2004 Elsevier Ltd. All rights reserved