Extended-spectrum beta-lactamase-producing Serratia marcescens outbreak in a Bulgarian hospital.

We describe a nosocomial outbreak of extended-spectrum beta-lactamase (ESBL)-producing Serratia marcescens in a Bulgarian university hospital affecting nine patients on four wards. Phenotypic and genotypic (plasmid profile, randomly amplified polymorphic DNA analysis and amplified ribosomal DNA restriction analysis) analysis of the isolates indicated a single clone. The epidemic strain was resistant to oxyimino beta-lactams, aztreonam, aminoglycosides, tetracycline and chloramphenicol. It produced CTX-M-3 ESBL as demonstrated by isoelectric focusing, CTX-M PCR-RFLP and gene sequencing. The isolate was also found in the environment and from a nurse's hands, suggesting transmission by staff handling. The outbreak was controlled by patient isolation and intensified hand washing. This is the first report from Bulgaria describing a hospital outbreak caused by CTX-M-3 ESBL-producing S. marcescens.

Nosocomial bloodstream infections with Burkholderia stabilis.

Burkholderia stabilis was grown from blood cultures of seven patients presenting with signs and symptoms of septicaemia in the intensive care unit at Mersin University Hospital, Mersin, Turkey between July and October 2002. Four patients had one B. stabilis-positive blood culture, two patients had two, and one patient had four. Isolates from six of seven patients had the same resistotype and random amplified polymorphic DNA analysis type. Despite treatment with ciprofloxacin and imipenem, to which the strains were susceptible, all patients died one to eight days after isolation of B. stabilis from their blood. B. stabilis should be regarded as an opportunistic pathogen that may cause nosocomial bloodstream infections.

Bacteriostatic and bactericidal activity of penicillins at constant and variable concentrations.

Temocillin, the first beta-lactamase stable penicillin which is active against Gram-negative bacteria, was shown to be much more potent than ampicillin, piperacillin and mezlocillin against most species of Enterobacteriaceae. All isolates of Escherichia coli producing TEM 1, TEM 2 or OXA 1 beta-lactamases were sensitive to temocillin. When the prolonged kinetics of elimination of temocillin were simulated in an in vitro system, more rapid killing and less regrowth occurred as compared with penicillins with more rapid elimination.

Sulbactam/ampicillin versus cefotaxime as initial therapy in serious soft tissue, joint and bone infections.

In an open, randomised comparative study, 23 patients with bone, joint or soft tissue infections were treated with ampicillin 2g plus sulbactam 1g 3 times a day or cefotaxime 2g 3 times a day as an initial 2-week therapy. Monoinfections with Staphylococcus aureus were the most common bone or joint infections. Clinical cure or improvement 2 weeks after the end of therapy was observed in all 13 patients treated with sulbactam/ampicillin and in 7 of the 8 patients evaluated for efficacy after treatment with cefotaxime. Most organisms identified before the onset of therapy were susceptible to the antibiotic randomly selected for therapy, although the majority of infections due to beta-lactamase-producing staphylococci could not have been treated with ampicillin without sulbactam. Treatment failed to eradicate S. aureus in 1 patient from each group. In addition, S. aureus infection recurred in 2 patients in the cefotaxime group within 2 weeks after the end of therapy. No serious side effects were observed.

Biliary concentrations of temocillin.

Serum and bile concentrations of temocillin were measured after intravenous administration of 1g twice daily to 16 patients with biliary tract infections. The concentration of temocillin in the bile showed high inter-individual variations; however, in patients drug concentrations were attained which were considerably higher than the minimum inhibitory concentrations of the biliary tract pathogens identified throughout the study period. All the patients admitted to the study were treated successfully by temocillin, with the tolerance of the drug being very good, and no side effects or drug-related deteriorations of laboratory data being reported.

Temocillin treatment of gynaecological infections with special reference to blood and tissue concentrations.

Tissue concentrations of temocillin were determined in samples from gynaecological surgical patients. Measurable concentrations of temocillin were observed during the entire time period investigated (up to 7 hours post administration). Inhibitory concentrations for the majority of susceptible bacteria were achieved. The therapeutic results observed in 40 patients with various infections (90% fully effective, 5% partially effective) confirm the high efficacy of temocillin in the treatment of gynaecological infections.

Decrease in nosocomial pneumonia in ventilated patients by selective oropharyngeal decontamination (SOD).

OBJECTIVE: To determine the influence of selective oropharyngeal decontamination (SOD) on the rate of colonization and infection of the respiratory tract in intensive care patients requiring mechanical ventilation for more than 4 days. A financial assessment was also performed. DESIGN: Randomized, prospective, controlled study using amphotericin B, colistin sulfate (polymyxin E), and tobramycin applied to the oropharynx and systemic cefotaxime prophylaxis. SETTING: Anesthesiology intensive care unit (ICU) of a 1500-bed hospital. PATIENTS: A total of 88 patients admitted as emergencies and intubated within less than 24 h were enrolled. Fifty-eight patients received SOD and 30 patients served as controls. Randomization was in the proportion of 2 : 1 study patients to controls. INTERVENTIONS: Microbiological samples from the oropharynx and other infected sites were taken at the time of admission, then twice a week and after extubation. MEASUREMENTS AND RESULTS: With the use of SOD, colonization was significantly reduced. Furthermore, the infection rate decreased from 77% in the controls to 22% in the study patients. Staphylococcus aureus was the main potential pathogen causing colonization and pneumonia. Number of days in the ICU, duration of ventilation, and mortality were not significantly decreased. The total cost of antibiotics was reduced. Development of resistance was not observed. CONCLUSIONS: The use of SOD significantly reduced the colonization and pneumonia and the total charge for antibiotics. The length of stay in the ICU, duration of ventilation, and mortality were similar. No resistance was observed. Staphylococcus aureus was selected by SOD in some patients and the clinical relevance needs further observation.

Comparative antimicrobial spectrum and activity of ceftibuten against clinical isolates from West Germany.

The in vitro activity of a new oral cephalosporin, ceftibuten, was determined against 837 clinical isolates by agar dilution technique and compared with that of the oral cephalosporins, cefaclor, cefuroxime, cefixime, cefpodoxime, and cefprozil. Against Enterobacteriaceae, ceftibuten was the most active of the compounds. Ceftibuten MIC90s were less than or equal to 0.25 micrograms/ml for most members of the family Enterobacteriaceae, 0.13 microgram/ml for Haemophilus influenzae, 4 micrograms/ml for Moraxella catarrhalis, and 0.5 microgram/ml for Neisseria gonorrhoeae. Ceftibuten also was active against beta-haemolytic streptococci (serogroups A, C, and G) and penicillin-susceptible strains of Streptococcus pneumoniae (MIC90, 4 micrograms/ml), but was not active against Staphylococcus spp. or the anaerobic bacteria studied. Cefpodoxime and cefuroxime were the most active of the cephalosporins against nonenteric streptococci; cefprozil and cefuroxime were the most active against staphylococci, and cefaclor demonstrated the greatest activity against some Bacteroides spp. Most strains of Acinetobacter baumanii, Pseudomonas spp., and methicillin-resistant staphylococci, as well as all strains of Clostridium difficile, were resistant to each of the cephalosporins tested.

Ceftibuten and bactericidal kinetics. Comparative in vitro activity against Enterobacteriaceae producing extended spectrum beta-lactamases.

Ceftibuten, compared to cefixime, cefetamet, cefpodoxime, loracarbef, cefprozil, cefuroxime, cefaclor, and cefadroxil, was the most active oral cephalosporin derivative against Enterobacteriaceae producing plasmid-encoded broad spectrum beta-lactamases. In a pharmacodynamic model, ceftibuten was bactericidal for Haemophilus influenzae and Streptococcus pneumoniae at concentrations simulating human serum levels following 200 mg, p.o., b.i.d.

A noncomparative study of cefprozil at two dose levels in the treatment of acute uncomplicated bacterial sinusitis.

The efficacy and safety of cefprozil at two dose levels were evaluated in 110 patients with acute uncomplicated bacterial sinusitis in an uncontrolled, noncomparative, Phase II trial. Ninety patients received 250 mg of cefprozil (low-dose group) and 20 patients received 500 mg of cefprozil (high-dose group) every 12 hours for ten days. Evaluable patients had symptoms consistent with acute sinusitis, pathogens isolated at pretreatment susceptible to cefprozil, and a radiograph positive for sinusitis within 48 hours before treatment. A satisfactory clinical response was achieved in 34 of 39 evaluable patients (87%) in the low-dose group and in all 16 evaluable patients (100%) in the high-dose group. Pathogens were eradicated in 35 of 39 patients (90%) in the low-dose group and in 15 of 16 patients (94%) in the high-dose group. A total of 140 of 155 pathogens (90%) isolated pretreatment were susceptible to cefprozil. Six patients (7%) in the low-dose group and one patient (5%) in the high-dose group reported at least one adverse clinical event.

Extended-spectrum beta-lactamase (ESBL) CTX-M-15-producing Escherichia coli and Klebsiella pneumoniae in Sofia, Bulgaria.

During a survey of extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae in Bulgaria in 2001-2002, three isolates from Sofia (two Escherichia coli, one Klebsiella pneumoniae) showed cefotaxime MICs that were decreased in the presence of clavulanate and were 2-8-fold higher than those of ceftazidime. Resistance was transferred to a sensitive recipient strain of E. coli. Both wild-type and transconjugant strains produced a cefotaxime-hydrolysing beta-lactamase of pI 8.8. Sequencing of the PCR product obtained with oligonucleotide primers binding outside the coding region identified this beta-lactamase as CTX-M-15. To our knowledge, this is the first report of CTX-M-15 in Bulgaria.

The influence of sulbactam on the in vitro activity of mezlocillin, piperacillin and cefotaxime.

In a multicentre study, the in-vitro activity of mezlocillin (MEZ, Chemical Abstract Service [CAS] 51481-65-3), piperacillin (PIP, CAS 61477-96-1) and cefotaxime (CTX, CAS 63527-52-6) against mezlocillin-resistant organisms was determined alone and in combination with the beta-lactamase inhibitor sulbactam (SBT, CAS 68373-14-8). A total of 870 strains were investigated (481 Enterobacteriaceae, 57 Pseudomonas aeruginosa, 41 Acinetobacter spp., 194 Bacteroides fragilis and 97 Staphylococcus spp.). MIC values were determined using the agar dilution test (aerobic organisms) or the microbroth dilution test (Bacteroides spp.) in accordance with Deutsche Industrie für Normung 58 940. SBT was added in fixed concentrations of 5 mg/l and 10 mg/l. For all combinations with SBT investigated, the geometric mean of the MIC and the MIC(50) and MIC(90) values were reduced as compared with the antibiotic alone (without SBT). Consequently, the proportion of sensitive strains was appreciably increased, for example in the Enterobacteriaceae: MEZ 1%, MEZ + 10 mg/l SBT 53%; PIP 4%, PIP + 10 mg/l SBT 54%; CTX 52%, CTX + 10 mg/l SBT 68%. The effect of SBT was especially pronounced on Bacteroides spp. For this organism, the proportion of sensitive strain rose from 2% to 97% (MEZ), 6% to 95% (PIP) and from 7% to 98% (CTX). The results show that adding SBT appreciably enhances the activity of MEZ, PIP and CTX against resistant strains of microorganism, and extends the activity spectrum to include anaerobic organisms. Thus the availability of SBT as a single-agent preparation for use in combination with various beta-lacta antibiotics represents a worthwhile enlargement of the therapeutic armamentarium for treating bacterial infections.

Epidemiological analysis of the spread of pathogens from a urological ward using genotypic, phenotypic and clinical parameters.

Surveillance studies using molecular typing methods help clinicians assess the rate of potential spread of pathogens. The rate of cross transmission of uropathogens among patients on a urological ward was investigated. Urine samples were collected from 144 patients with urinary catheters and a significant bacteriuria. In a subgroup of 54 of these patients, cultures from a rectal swab were also made. Typing by PFGE, RAPD or bacteriocins showed that 41% of uropathogens were related and represented by 38 typing patterns. Endogenous infection was present in 30% and exogenous infection in 38% of isolates. Altogether, there was a high rate of clonal relationship amongst uropathogens in our urological ward and we conclude that hygienic means and measures are far from being optimal.

Placebo-controlled, double-blind, randomized study of aerosolized tobramycin for early treatment of Pseudomonas aeruginosa colonization in cystic fibrosis.

In chronic Pseudomonas aeruginosa pulmonary infection of patients with cystic fibrosis (CF), antibiotic therapy generally fails to eradicate the bacterial pathogen. The mucoid bacterial phenotype, high sputum production by the host, and low airway levels of antibiotics seem to be responsible for the observed decrease in antibiotic efficacy. We hypothesized that early antibiotic treatment by inhalation in CF patients may be able to prevent or at least delay airway infection. In a prospective placebo-controlled, double-blind, randomized multicenter study, 22 CF patients received either 80 mg b.i.d. of aerosolized tobramycin or placebo for a period of 12 months shortly after the onset of P. aeruginosa pulmonary colonization. Two patients in the tobramycin and six patients in the placebo group stopped inhalation before the 12 month treatment period. Using life table analysis, the time to conversion from a P. aeruginosa-positive to a P. aeruginosa-negative respiratory culture was significantly shorter in the tobramycin-treated group than in the placebo group (P < 0.05, log rank test). Lung function parameters and markers of inflammation did not change in either group during treatment. The results of this study suggest that early tobramycin inhalation may prevent and/or delay P. aeruginosa pulmonary infection in CF patients.

RU 29 246, the active compound of the cephalosporin-prodrug-ester HR 916. I. Antibacterial activity in vitro.

The aminothiazolyl-cephalosporin RU 29 246 is the active metabolite of the prodrug-pivaloyl-oxyethyl-ester HR 916. RU 29 246 in vitro activity includes a wide range of clinically relevant bacterial pathogens. Against methicillin-sensitive Staphylococci RU 29 246 (MIC90 of 0.25 approximately 2 micrograms/ml) was clearly more active than cefaclor, cefuroxime, cefpodoxime, cefixime and ceftibuten, but slightly less active than cefdinir. RU 29 246 inhibited hemolytic Streptococci of the serogroups A, B, C and G as well as penicillin-sensitive Streptococcus pneumoniae at concentrations similar to cefdinir, cefpodoxime and cefuroxime (MIC90 less than or equal to 0.13 micrograms/ml), but less than the other oral cephalosporins investigated (cefixime, cefaclor and ceftibuten). MIC90s of RU 29 246 against Escherichia coli, Klebsiella pneumoniae, Klebsiella oxytoca, Salmonella spp., Shigella spp., Proteus mirabilis and Haemophilus influenzae were less than or equal to 0.5 micrograms/ml. Only RU 29 246 and cefdinir demonstrated moderate activity against Acinetobacter baumannii (MIC90 greater than or equal to 4 micrograms/ml). Most strains of Pseudomonas spp., Serratia marcescens, Enterobacter spp., Hafnia alvei and Bacteroides spp. were resistant to RU 29 246. RU 29 246 killed Escherichia coli and Staphylococcus aureus at a rate of 99% to 99.9% at concentrations of two times MIC. The pH value of the medium (range 5.5 to 8.5) and the inoculum size (range 10(5) to 10(7) cfu/ml) had no or only low influence on the antibacterial activity of RU 29 246. RU 29 246 is a broad spectrum cephalosporin including in its activity both Gram-positive and Gram-negative pathogens and therefore--depending on the bioavailability of its prodrug--looks promising as to its therapeutic perspective.

RU 29 246, the active compound of the cephalosporin-prodrug-ester HR 916. II. Stability to beta-lactamases and affinity for penicillin-binding proteins.

The aminothiazolyl-cephalosporin RU 29 246, the active metabolite of the prodrug-ester HR 916, is active against strains producing the widespread plasmid-encoded TEM-1, TEM-2 and SHV-1 beta-lactamases. Except for TEM-7 the activity of RU 29 246 against strains producing extended broad spectrum beta-lactamases (TEM-3, TEM-5, TEM-6, SHV-2, SHV-4, SHV-5, CMY-1, CTX-M), however, is low. Relative hydrolysis rates of RU 29 246 are comparable with those of cefpodoxime, the active metabolite of CS-807, and are extremely low for the TEM-1 and SHV-1 beta-lactamases. The compound demonstrates remarkable inhibitory activity against the chromosomal beta-lactamase of Enterobacter cloacae P99. In the presence of 1.7 microM this enzyme loses 50% of its activity. At concentrations of 0.43, 0.003 and 0.01 micrograms/ml the compound binds preferentially to the penicillin-binding protein (PBP) 3 of Escherichia coli K12, to the PBPs 2x and 3 of Streptococcus pneumoniae R6 and to PBP 1 of Staphylococcus aureus SG 511, respectively.

Plasmid-encoded AmpC beta-lactamases: how far have we gone 10 years after the discovery?

The dogma that ampC genes are located exclusively on the chromosome was dominant until about 10 years ago. Since 1989 over 15 different plasmid-encoded AmpC beta-lactamases have been reported from several countries. Most of these enzymes evolved in two clusters. The major cluster includes several enzymes with a high similarity to CMY-2, which is the closest related chromosomal AmpC enzyme of Citrobacter freundii. A second cluster centers around CMY-1. It is less homogeneous and not closely related chromosomal AmpC enzymes. Molecular diversification by amino acid substitutions does not usually translate into a change in the resistance phenotype. At this time, CMY-2 appears to be the most prevalent and widely distributed. Further global increase of prevalence and diversity of plasmidic AmpC beta-lactamases have to be anticipated in the next millenium.

PBP binding and periplasmic concentration as determinants of the antibacterial activities of three new oral cephalosporins in Escherichia coli.

The antibacterial activities of cefetamet, cefixime and cefuroxime were investigated in Escherichia coli with regard to their penetration rates through the outer membrane and their affinities for PBPs in Escherichia coli. The permeability coefficient of cefetamet was measured in E. coli C600 carrying a pUC18 plasmid derivative, in which the gene of a transferable cephamycinase (CMY-2) was cloned, whereas diffusion of cefixime and cefuroxime was measured in E. coli C600 harbouring the OXA-1 beta-lactamase gene. It was found that cefetamet penetrated 5 and 9 times faster than cefixime and cefuroxime, respectively. The correlation between antibacterial activities and PBP affinities was studied in E. coli C600 not producing beta-lactamases. In this strain, cefetamet and cefixime shared the same inhibitory activity (MIC = 1 microgram/ml for both antibiotics) and the same affinity for PBP 3 (ID50 = 0.25 micrograms/ml). Cefuroxime had a lower inhibitory activity (MIC = 4 micrograms/ml) and a lower affinity for PBP 3 (ID50 = 0.5 microgram/ml). Cefixime and cefuroxime had 20 and 10 times higher affinity, respectively, than cefetamet for PBP 1s. It was concluded that the superior PBP affinity of cefixime can counterbalance the better penetration of cefetamet through the outer membrane, whilst differences in either permeability or PBP affinity seem sufficient to explain the lower antibacterial activity of cefuroxime compared to cefixime, which might well be related to the poor stability of the cefuroxime molecule.

[Imipenem resistance in Pseudomonas aeruginosa]. / Imipenem-Resistenz bei Pseudomonas aeruginosa.

In 1997 in western Austria, 9.9% of Pseudomonas aeruginosa strains from patients of general practitioners were resistant to imipenem as well as 18.2% of the isolates from hospitals and 20.2% of the strains at a university teaching hospital. Within the hospital the imipenem resistance varied from 9.9% among out-patients to 28.7% in isolates from intensive care units. In medical/surgical words, up to 15.1% of P. aeruginosa strains were resistant to imipenem. The incidence of imipenem-resistant P. aeruginosa strains correlates to the use of carbapenems. In June 1997, 10 consecutive isolates from 8 patients were obtained and typed using restriction fragment length polymorphism analysis (RFLP) and Pyocin typing. All 10 isolates were resistant to meropenem as well as to imipenem. The finding (by RFLP and Pyocin typing) of individual bacterial types in each isolate strongly contradicts the spread of infection by cross infection. However, all patients were proven to have been treated with imipenem during the 3 months prior to testing. In 1997, 13,880 g of imipenem were used at the university hospital in Innsbruck. The use of carbapenems appears to be the main cause for the increased incidence of imipenem-resistant P. aeruginosa strains.

[Intragastric formation of ammonia in Campylobacter pylori associated gastritis. Diagnostic and pathogenetic significance]. / Intragastrale Ammoniakbildung bei Campylobacter-pylori-assoziierter Gastritis. Diagnostische und pathogenetische Bedeutung.

The diagnostic relevance of measurement of ammonia (NH3) in not stimulated gastric juice in patients with campylobacter pylori associated gastritis (CPAG) is in discussion. The role of CP-urease induced NH3 in pathogenesis of active gastritis is unclear. In answering to this questions we evaluated the sensitivity and specifity of NH3-test and CLO-test in cases of CPAG (n = 50), non CPAG (n = 16) and normal gastric mucosa (n = 20). We found a 88% sensitivity and a 86% specifity for NH3-test, a sensitivity for CLO-test of 80% and a specifity rate of 87%. NH3-test correlated well with CLO-test (n = 51, p less than 0.01) and semiquantitative histological identification of CP (p less than 0.01, n = 22). On the other hand we tried to correlate the amount of NH3 in the gastric juice and the histological degree of gastritis activity (infiltration of leucocytes of the lamia propria) in CPAG (n = 78) and Non-CPAG (n = 32) and before and after therapy in CPAG (n = 9) with bismuthsubnitrate (2 g/d, 14 d). There was no correlation between the amount of NH3 and the degree of active chronic gastritis in patients with CPAG (with or without therapy) and patients with non CPAG. It seems that NH3 has a diagnostic but no pathogenetic role in the process of inflammatory activity of CPAG.