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Effect of androgens and glucocorticoids on microbial growth and antimicrobial susceptibility.

The effects of androgens, testosterone and dihydrotestosterone (DHT), of an environmental anti-androgen, 2,2-bis(4-chlorophenyl)-1,1-dichloroethylene (DDE), and of glucocorticoids, hydrocortisone and dexamethasone, on growth kinetics and antibiotic susceptibility of E. faecalis, E. coli, P. aeurginosa, and S. aureus were measured. For P. aeurginosa, the presence of either DHT or DDE caused at least a fourfold shift in the minimum inhibitory concentration (MIC) of cefepime and tobramycin. DHT and DDE also affected the response of E. faecalis to meropenem and norfloxacin, resulting in a shift from sensitive to intermediate resistance (four-fold increase in MIC). Hydrocortisone (2 microM) induced an increase in the sensitivity of S. aureus to erythromycin, as compared to hormone-free control (from 0.5 to 0.06 microg/mL). The susceptibility pattern of E. coli was unaffected by the hormones tested. These changes in susceptibility to antibiotics were unrelated to alterations in growth kinetics. For all organisms tested, the alterations in MICs occurred only in the presence of hormone, indicative of changes in the phenotype of these stable quality control strains.

Androgens↗

Agrobacterium-mediated transformation of protocorm-like bodies in Cymbidium.

Genetically transformed plants of Cymbidium were regenerated after cocultivating protocorm-like bodies (PLB) with Agrobacterium tumefaciens strain EHA101 (pIG121Hm) that harbored genes for beta-glucuronidase (gus), hygromycin phosphotransferase (hpt) and neomycin phosphotransferase II (nptII). PLB of three genotypes maintained in liquid new Dogashima medium (NDM), were subjected to transformation experiments. The PLB inoculated with Agrobacterium produced secondary PLB, 4 weeks after transfer onto 2.5 g L(-1) gellan gum-solidified NDM containing 10 g L(-1) sucrose, 20 mg L(-1) hygromycin and 40 mg L(-1) meropenem. Transformation efficiency was affected by genotype and the presence of acetosyringone during cocultivation. The highest transformation efficiency was obtained when PLB from the genotype L4 were infected and cocultivated with Agrobacterium on medium containing 100 muM acetosyringone. Transformation of the hygromycin-resistant plantlets regenerated from different sites of inoculated PLB was confirmed by histochemical GUS assay, PCR analysis and Southern blot hybridization.

Base Sequence↗

Listeria meningitis in transplant recipients.

INTRODUCTION: Meningitis is a rare complication following organ and stem-cell transplantation and can be caused by a variety of microorganisms. AIM: To retrospectively review the clinical course and outcome of five cases of listeriosis in four organ recipients and one stem-cell recipient during a seven-year period. PATIENTS AND METHODS: Patient records for more than 3500 patients undergoing organ or stem-cell transplantation at the university hospital of Innsbruck during a 27-year period were evaluated. Standard immunosuppression consisted of calcineurin inhibitor-based triple drug therapy with or without ATG or IL2 receptor antagonist induction. RESULTS: The first case affected a 35-year-old woman who received an allogenic bone marrow transplant for advanced breast cancer. Cases two and three related to two male heart recipients. Cases four and five were diagnosed in one male and one female renal recipient. Listeria monocytogenes was isolated from blood in two cases and from cerebrospinal fluid in three. Treatment consisted of ampicillin in all cases with the addition of tobramycin (1), TMPS (1), meropenem (2) or imipenem/cilastatin (1). The deaths of two patients were directly related to L. monocytogenes. CONCLUSIONS: Although listeriosis is a rare complication following transplantation, this infection should be ruled out in individuals presenting with neurological symptoms and fever.

Adult↗

Piperacillin-tazobactam is more effective than ceftriaxone plus gentamicin in febrile neutropenic patients with hematological malignancies: a randomized comparison.

GOALS: Efficacy and costs of empirical antibacterial therapy in febrile neutropenic patients are important issues. Several strategies have been reported to be similarly effective: monotherapy with cefepime, ceftazidime or a carbapenem or duotherapy with an antipseudomonal beta-lactam antibiotic or ceftriaxone in combination with an aminoglycoside. Piperacillin-tazobactam monotherapy is promising, but its role in this setting still has to be defined. PATIENTS AND METHODS: Of 212 consecutive febrile episodes in 130 neutropenic patients with hematological malignancies randomized to receive either piperacillin-tazobactam (4.5 g every 8 h; group A) or ceftriaxone (2 g once daily plus gentamicin 5 mg/kg once daily; group B), 183 episodes (98 group A, 85 group B) were evaluable for response. RESULTS: Defervescence within 72 h without modification of the antibiotic therapy was achieved in 56/98 episodes (57.1%) in group A and in 30/85 (35.3%) in group B (P=0.0047). If fever persisted, teicoplanin plus gentamicin (group A) or teicoplanin plus ciprofloxacin (group B) were added. All patients still febrile then received meropenem, teicoplanin and amphotericin B. With these modifications of antibiotic therapy, 89.8% of patients in group A had responded at 21 days but only 71.8% in group B (P=0.005). The mean total antibiotic drug cost in group A was only 39.4% of that in group B (euro 445 versus euro 1129; P=0.010). CONCLUSION: Piperacillin-tazobactam monotherapy is significantly more effective and cost-efficient than ceftriaxone plus gentamicin as first-line therapy in febrile neutropenic patients with hematological malignancies.

Adolescent↗

Drug susceptibility of isolates from severe postoperative intraperitoneal infections causing multiple organ failure.

PURPOSE: To select the most appropriate antibiotic regimens for life-threatening postoperative infections, we obtained isolates from patients with severe postoperative infections over a 12-year-period, and examined their drug susceptibility. METHODS: The subjects of this study were 55 patients with multiple organ failure (MOF) caused by postoperative infection. RESULTS: All strains of Methicillin-resistant Staphylococcus aureus (MRSA) were susceptible to Vancomycin (VCM) and Teicoplanin (TEIC). Only 0.3% of all the Pseudomonas aeruginosa strains were resistant to Imipenem (IPM), but 53.6% of the strains from the severe infections were resistant to IPM. On the other hand, there were few P. aeruginosa strains resistant to Meropenem (MEPM), Ceftazidime (CAZ), Ciprofloxacin (CPFX), and Pazufloxacin (PZFX), even among strains isolated from severe infections. The resistant rate of Bacteroides fragilis to Clindamycin (CLDM) was 35.9%, but there were strains resistant to IPM and Panipenem. CONCLUSION: These findings suggest that VCM or TEIC are most appropriate for severe abdominal abscess caused by MRSA, whereas MEPM, CAZ, CPFX, and PZFX are more effective against P. aeruginosa infections. The only antibiotic effective against B. fragilis infections in this study was IPM.

Abdominal Abscess↗

[Soft tissue infections in oral, maxillofacial, and plastic surgery. Bacterial spectra and antibiotics].

BACKGROUND: Soft tissue infections in the maxillofacial region are mainly caused by Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli but also by members of the genera Enterococcus, Klebsiella, and Enterobacter, respectively. METHODS: In a prospective study 96 patients with severe maxillofacial non-odontogenic postoperative soft tissue infections were analyzed with regard to the bacterial spectrum and resistance patterns against antibiotics. The dominating bacteria were Streptococci (25% of the isolates) and Staphylococci (24% of the isolates). In addition, members of Enterobacteriaceae were isolated in approximately 10% of the cases. The most frequent anaerobes found were as follows: Peptostreptococcus, Eubacterium, Prevotella and Fusobacterium. RESULTS: The resistance rates against antibiotics found were: penicillin G 36%, ampicillin 42%, and doxycycline 36%. In addition, the resistance rate against erythromycin and clindamycin was 26% and 7%, respectively. No resistant strains were detected against vancomycin and teicoplanin. All anaerobes showed a low antimicrobial resistance as previously described for odontogenic infections. CONCLUSION: In summary, soft tissue infections in the maxillofacial region present a different spectrum of bacteria in contrast to the well-investigated odontogenic infections. Antibiotic administration should be, whenever possible, performed after differentiation of involved strains and resistogram. The most promising antibiotics are imipenem, meropenem, and ciprofloxacin as well as cefotiam. Modern fluoroquinolones will be the antibiotics of the future.

Adolescent↗

Emergence of cotrimoxazole- and quinolone-resistant Campylobacter infections in bone marrow transplant recipients.

Clinical and microbiological data were collected prospectively from 704 patients who underwent bone marrow transplantation (BMT) during an 11-year period (1991-2001), and the first two cases of Campylobacter infection occurring in BMT recipients in the pre-engraftment period were identified. The two cases occurred on days 2 and 3 post-BMT, respectively. Both patients had Campylobacter jejuni enteritis, and one case was complicated by bacteraemia. In both cases the presenting symptoms were indistinguishable from hospital-acquired pre-engraftment diarrhoea, which is commonly caused by Clostridium difficile. Both of the Campylobacter jejuni isolates were resistant to cotrimoxazole and ciprofloxacin. Both patients responded to intravenous meropenem and subsequently had uneventful marrow engraftment.

Adolescent↗

Use of the E test to assess synergy of antibiotic combinations against isolates of Burkholderia cepacia-complex from patients with cystic fibrosis.

Treatment of Burkholderia cepacia-complex infections in cystic fibrosis patients is problematic, since the microorganism is often resistant to most antimicrobial agents. In this study, the Epsilometer test, or E test, was used to assess the activity of antimicrobial combinations against Burkholderia cepacia-complex. In a preliminary evaluation, the E test was compared to the checkerboard method using 10 test organisms. Synergy testing by the E test was then performed on 131 clinical isolates of Burkholderia cepacia-complex using various combinations of antimicrobial agents. Agreement between the E test and the checkerboard method was 90%. The rate of resistance to individual agents ranged from 48% for meropenem to 100% for tobramycin, chloramphenicol, and rifampin. In 71.6%, 15.6%, and 12.6% of the test evaluations performed, the combinations tested resulted in additivity/indifference, synergism, and antagonism, respectively. The highest rates of synergy were observed with combinations of ciprofloxacin-piperacillin (44%), rifampin-ceftazidime (33%), chloramphenicol-ceftazidime (22%), cotrimoxazole-piperacillin/tazobactam (22%), and ciprofloxacin-ceftazidime (21%). Rates of antagonism for cotrimoxazole and chloramphenicol in combination with beta-lactam agents were higher than those observed for ciprofloxacin plus beta-lactam agents. These results suggest that the E test is a valuable and practical method to be considered for improving the identification of possible therapeutic options in cystic fibrosis patients infected with organisms belonging to the Burkholderia cepacia-complex.

Bacteriological Techniques↗

Comparison of the Micronaut Merlin automated broth microtiter system with the standard agar dilution method for antimicrobial susceptibility testing of mucoid and nonmucoid Pseudomonas aeruginosa isolates from cystic fibrosis patients.

The aim of this study was to compare a commercially available automated broth microdilution system (Merlin; Micronaut, Germany) with the standard agar dilution method for susceptibility testing of pulmonary isolates from cystic fibrosis patients. Accurate susceptibility testing of bacterial isolates from cystic fibrosis patients is known to pose problems. Although commercially available automated test systems could facilitate susceptibility testing of such isolates in routine diagnostics, these systems have not been recommended thus far. However, a pilot study recently indicated that the Merlin system, which is based on an endpoint measurement rather than on growth curves, might be applicable in the susceptibility testing of isolates from cystic fibrosis patients. In the present study, the Merlin system was further evaluated using an extended panel of nonmucoid and mucoid Pseudomonas aeruginosa isolates. The results showed that the MICs obtained with the Merlin system tended to be lower than those obtained with the agar dilution method, a finding that became increasingly apparent when mucoid Pseudomonas aeruginosa strains were tested. The correlation coefficients (r values) of the MIC results for all strains tested were between 0.6 and 0.8 for five of the seven antimicrobial agents, with r values exceeding 0.8 for only meropenem and ciprofloxacin. However, since the overall rate of serious discrepancies was within an acceptable range, the Merlin system appears to be applicable for routine clinical use in susceptibility testing of P. aeruginosa isolates from cystic fibrosis patients.

Anti-Bacterial Agents↗

Pneumonia involving Aspergillus and Rhizopus spp. after a near-drowning incident with subsequent Nocardia cyriacigeorgici and N. farcinica coinfection as a late complication.

Reported here is the case of a 22-year-old man who developed pneumonia with unusual pathogens after a near-drowning incident. On day 7 following admission, Rhizopus spp. and Aspergillus fumigatus were cultured from the patient's bronchoalveolar lavage fluid. One week later, sputum cultures revealed N. cyriacigeorgici as well as N. farcinica. The patient recovered fully after prolonged therapy with liposomal amphotericin B, amikacin, meropenem and cotrimoxazole.

Adult↗

Effect of oxygen limitation on the in vitro antimicrobial susceptibility of clinical isolates of Pseudomonas aeruginosa grown planktonically and as biofilms.

Pseudomonas aeruginosa, the predominant causative pathogen of chronic lung infection in patients with cystic fibrosis, may grow under anaerobic conditions as biofilms in the lungs of cystic fibrosis patients. To determine if growth under anaerobic conditions affects the antimicrobial susceptibility of P. aeruginosa, the susceptibility of clinical isolates of P. aeruginosa grown planktonically and as biofilms to a range of antibiotics was determined. Growth under anaerobic conditions did not reduce the ability of ceftazidime, meropenem, aztreonam, piperacillin, or piperacillin/tazobactam to inhibit planktonic growth, with MIC50 values for these antibiotics remaining unchanged or decreasing. However, tobramycin was less effective at inhibiting planktonic bacterial growth under anaerobic conditions, with the MIC50 of tobramycin increasing twofold. Growth under anaerobic conditions also decreased the bactericidal activity of tobramycin, with the MBC50 of tobramycin increasing fourfold. The killing kinetics of tobramycin was also examined under aerobic and anaerobic conditions for selected isolates. When isolates 6A and 12A were grown aerobically, concentration-dependent decreases in total viable count were apparent with tobramycin. In contrast, when these isolates were grown anaerobically, tobramycin at the same concentrations did not decrease the total viable count. When isolates were grown as biofilms under both aerobic and anaerobic conditions, isolate- and concentration-dependent differences in killing of the biofilms by tobramycin were apparent. However, tobramycin at concentrations up to 128 mg/l was unable to eradicate biofilms of any of the isolates tested, whether biofilms were grown aerobically or anaerobically. These results show that oxygen limitation may reduce, in a strain-dependent manner, the susceptibility to tobramycin of P. aeruginosa grown planktonically and as biofilms.

Aerobiosis↗

Evolution of resistance among clinical isolates of Acinetobacter over a 6-year period.

The aim of this report was to study the evolution of susceptibilities of 1532 clinical isolates of Acinetobacter recovered over a period of 6 years. The minimal inhibitory concentrations (MICs) of 15 antimicrobial agents were determined for all the isolates. The respective percentages of resistant strains in the years 1991 and 1996 were as follows: ciprofloxacin, 54.4% and 90.4%; tobramycin, 33% and 71.8%; amikacin, 21% and 83.7%; ampicillin plus sulbactam, 65.7% and 84.1%; ceftazidime, 57.4% and 86.8%; ticarcillin, 70% and 89.4%; trimethoprim plus sulfamethoxazole, 41.1% and 88.9%; and imipenem, 1.3% and 80%. The MIC90s of ciprofloxacin, sparfloxacin, biapenem, meropenem, imipenem, cefepime, cefpirome, and rifampicin against 250 imipenem-resistant Acinetobacter strains were >32, >32, 128, >256, 256, >256, 256, and 16 mg/l, respectively. With serious infections, it was necessary to resort to the use of colistin, the only antibiotic active in vitro.

Acinetobacter↗

Antimicrobial resistance in European isolates of Pseudomonas aeruginosa. European SENTRY Participants.

Pseudomonas aeruginosa is responsible for a substantial fraction of hospital infections. Twenty-five European university hospitals submitted a total of 1411 Pseudomonas aeruginosa isolates for susceptibility testing during 1997 and 1998. The isolates showed highest susceptibility to amikacin (87.5%), meropenem (87.3%) and piperacillin/tazobactam (86.8%). Susceptibility to ciprofloxacin was 73.2%. There was no clear geographical distribution of resistance, although isolates from northwestern Europe tended to be more susceptible than those from southeastern Europe. Isolates that were resistant to one class of antibiotics were also often resistant to at least one other class of antibiotics. Imipenem-resistant isolates were generally not clonally related.

Anti-Bacterial Agents↗

Frequency of isolation and antimicrobial resistance of gram-negative and gram-positive bacteria from patients in intensive care units of 25 European university hospitals participating in the European arm of the SENTRY Antimicrobial Surveillance Program 1997-1998.

A total of 3,981 isolates from patients treated at intensive care units were collected in 25 European university hospitals during 1997 and 1998 as part of the SENTRY Antimicrobial Surveillance Program. Overall, the most important species isolated were Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, coagulase-negative staphylococci (CNS), Enterobacter spp., Haemophilus influenzae, Streptococcus pneumoniae, and Enterococcus faecalis. Thirty-nine percent of all Staphylococcus aureus isolates were resistant to oxacillin. All Staphylococcus aureus isolates were fully susceptible to linezolid and vancomycin. Moreover, all CNS isolates were susceptible to vancomycin and minocycline. All Enterococcus faecalis isolates were susceptible to vancomycin, and 99% of these isolates were also susceptible to ampicillin. The antimicrobial agents most effective against Pseudomonas aeruginosa isolates were amikacin, piperacillin/tazobactam, meropenem, and cefepime, with 87, 85, 84, and 83% of isolates being susceptible, respectively. Escherichia coli isolates were fully susceptible to carbapenems, and at least 99% of these isolates were susceptible to ceftriaxone, cefepime, and amikacin. The Enterobacter spp. were also highly susceptible to carbapenems, amikacin, and cefepime, with 99, 97, and 96% of isolates being susceptible, respectively. Haemophilus influenzae was susceptible to most of the antibiotics tested. Only 68% of the pneumococcal isolates were fully susceptible to penicillin, yet 100% were susceptible to a number of fluoroquinolones and vancomycin. There are still sufficient treatment options for patients infected with the most important bacterial species involved in infections in intensive care units. However, the situation for patients with Pseudomonas aeruginosa infections is critical.

Anti-Bacterial Agents↗

Antimicrobial susceptibility of respiratory isolates of Enterobacteriaceae and Staphylococcus aureus in Italy: incidence and trends over the period 1997-1999.

The antibiotic susceptibility of members of the family Enterobacteriaceae and of Staphylococcus aureus strains isolated from the respiratory tract was assessed over the period 1997-1999 as part of the Italian Epidemiological Observatory survey sponsored by the Smith-Kline Foundation. A standardised method was used to determine the MICs of 22 antibiotics against isolates of Klebsiella pneumoniae (n=870), Escherichia coli (n=684), Enterobacter cloacae (n=342), Enterobacter aerogenes (n=187) and Serratia marcescens (n=135) as well as the MICs of 11 antibiotics against isolates of Staphylococcus aureus (n=1,606). Overall, the susceptibility rate of Enterobacteriaceae isolates was > or = 90% to 5 agents (meropenem, imipenem, amikacin, cefepime and gentamicin); 89-80% to 2 agents (ciprofloxacin and tobramycin); and <80% to 11 agents (cefotaxime, piperacillin-tazobactam, trimethoprim-sulfamethoxazole, cefetamet, ceftriaxone, ceftazidime, aztreonam, ticarcillin-clavulanate, tetracycline, piperacillin, cefuroxime, chloramphenicol, ticarcillin, amoxicillin-clavulanate and amoxicillin). During the 3-year monitoring period, antibiotic susceptibility increased in Klebsiella pneumoniae against amoxicillin-clavulanate, in Escherichia coli against third-generation cephalosporins and aztreonam, in Enterobacter aerogenes against amoxicillin and piperacillin-tazobactam and in Serratia marcescens against most of the antibiotics. In contrast, Enterobacter cloacae showed a tendency to develop resistance to cefetamet, amikacin and ciprofloxacin. Of the total number of Staphylococcus aureus strains, 38% were methicillin resistant. Nearly 80% of the methicillin-resistant strains displayed a multiresistance pattern (additional resistance to 2 or more non-beta-lactam antibiotics). Rates of susceptibility of particular species (Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus) were compared using strains from different geographical areas of Italy (northern, central and southern) and from different nosocomial areas (outpatients, intensive care unit [ICU] inpatients, non-ICU inpatients). Susceptibility of Klebsiella pneumoniae to several antibiotics was lower in southern Italy, whereas the incidence of methicillin-resistant strains was higher in northern and central Italy. The susceptibility of Escherichia coli was similar in all three areas. No significant differences in susceptibility of Klebsiella pneumoniae or Escherichia coli were found between strains from inpatients and outpatients or from inpatients admitted to ICU and non-ICU units. The incidence of methicillin-resistant Staphylococcus aureus was higher in ICU inpatients (52%) than in non-ICU inpatients (38%) and lower in outpatients (19%) than in inpatients.

Drug Resistance, Bacterial↗

Changes of antimicrobial resistance and extended-spectrum beta-lactamase production in Klebsiella spp. strains.

Changes of antimicrobial resistance and extended-spectrum beta-lactamase (ESBL) production of Klebsiella spp. strains isolated at the Central Military Hospital, Ruzomberok, Slovakia, with a special focus on the Anesthesiology--Resuscitation and Intensive Medicine Department during the years 1998-2002, were analyzed. Of 3920 gram-negative strains isolated from clinical materials during this period, Klebsiella spp. represented 8%. The incidence of ESBL-producing Klebsiella spp. isolates increased from 29% in 1998 to 69% in 2002. Of 17 antibiotics tested, meropenem was found to be the most effective drug (100%). The overall efficacy of cefotaxime was 31%, that of gentamicin 23%, and that of ciprofloxacin 54%. Analyzed Klebsiella isolates were characterized also by a high degree of multiresistance (53%). The high incidence of reduced antibiotic susceptibility among Klebsiella spp. strains isolated at the intensive-care department suggests that more effective strategies are necessary to control the selection and spread of resistant organisms in this hospital.

Anti-Bacterial Agents↗

In vitro activity of beta-lactams and quinolones against AmpC beta-lactamase-producing Escherichia coli.

We studied the antimicrobial susceptibility of AmpC beta-lactamase-producing Escherichia coli isolates collected at ten medical institutions in the Kinki area of Japan during a 6-month period (November 2002 through April 2003). Of 2845 E. coli isolates tested, 29 (1.0%) showed a minimum inhibitory concentration (MIC) for cefazolin of more than 8 microg/ml and were three-dimensional extract test positive. In standard inoculum susceptibility tests against these 29 strains, the MIC90s for the four carbapenems tested ranged from 0.06 microg/ml to 0.5 microg/ml, and these compounds were more active than the other beta-lactams, with meropenem being the most active. The MIC90s for beta-lactams, except carbapenems, ranged from 4 microg/ml to 32 microg/ml, with cefepime being the most active. In high inoculum susceptibility tests against these strains, the MIC90s for the four carbapenems and cefepime were 8 microg/ml or less, and these compounds were more active than other beta-lactams. The MIC90s for beta-lactams, except carbapenems and cefepime, were 32 microg/ml or more. The MIC90s for the five quinolones tested ranged from 4 microg/ml to 16 microg/ml, and the order of increasing susceptibility was ciprofloxacin > levofloxacin, gatifloxacin and pazufloxacin > prulifloxacin.

Anti-Bacterial Agents↗

Estimation of outer membrane permeability of carbapenem antibiotics to Pseudomonas aeruginosa.

The outer membrane permeability of carbapenems (imipenem [IPM], panipenem [PAPM], meropenem [MEPM], and biapenem [BIPM]) and ceftazidime (CAZ) to Pseudomonas aeruginosa was determined by the Zimmermann and Rosselet method. The permeability coefficients of beta-lactams tested at 50 &mgr;M concentration of substrates ranged from (0.40 +/- 0.10) x 10-6 cm/s for CAZ to (2.33 +/- 0.33) x 10-6 cm/s for IPM, indicating that the outer membrane permeability of carbapenems to P. aeruginosa was high in comparison with that of CAZ. In particular, IPM and BIPM showed a higher rate of penetration than MEPM and PAPM.

Journal Article↗