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PubMed2004

First identification of an Escherichia coli clinical isolate producing both metallo-beta-lactamase VIM-2 and extended-spectrum beta-lactamase IBC-1.

An Escherichia coli strain with decreased susceptibility to carbapenems was isolated from a hospitalised patient in Athens, Greece. The strain was resistant to all beta-lactams, including aztreonam, whereas the MIC of imipenem and meropenem was 0.5 mg/L. A positive EDTA-disk synergy test suggested the production of a metallo-beta-lactamase. PCR experiments revealed the presence of the bla(VIM-2), bla(IBC-1), and bla(TEM-1) genes. Resistance to beta-lactams was not transferable by conjugation. This is the first report of a clinical isolate of E. coli producing VIM-2, and the first report of the coexistence of bla(VIM-2) and bla(IBC-1) in a single clinical isolate.

Anti-Bacterial Agents↗

PubMed2005

IMPs, VIMs and SPMs: the diversity of metallo-beta-lactamases produced by carbapenem-resistant Pseudomonas aeruginosa in a Brazilian hospital.

Pseudomonas aeruginosa isolates (n=183), collected from bacteraemic patients hospitalised in Sao Paulo Hospital (Brazil) during 2000-2001, were screened for susceptibility to antimicrobial agents. The polymyxins were the most active compounds (100% susceptibility), followed by amikacin and cefepime (59.0%), meropenem (57.4%), and imipenem and gentamicin (55.2%). Imipenem-resistant isolates were ribotyped and screened for production of metallo-beta-lactamases (MBLs) by PCR with primers for bla(IMP), bla(VIM) and bla(SPM). MBL production was detected in 36 isolates (19.7% of the entire collection; 43.9% of the imipenem-resistant isolates) and the MBLs included SPM-1-like (55.6%), VIM-2-like (30.6%) and IMP-1-like (8.3%) enzymes.

Anti-Bacterial Agents↗

PubMed2005

An evidence-based evaluation of important aspects of empirical antibiotic therapy in febrile neutropenic patients.

Febrile neutropenia is still associated with a high mortality rate, making timely and efficient empirical antibiotic therapy absolutely vital. For these reasons, evidence-based guidelines are urgently needed. The guidelines published so far are mainly based on clinical experience and selective citation. This review summarises studies and meta-analyses concerning empirical antibiotic therapy in high-risk neutropenic patients: (1) No benefit results from the addition of an aminoglycoside to the initial empirical therapy. On the contrary, patients who received an aminoglycoside had a significantly higher rate of adverse events, especially nephrotoxicity. (2) The empirical addition of a glycopeptide after 3-4 days of persistent fever was evaluated in two randomised controlled trials. Combined analysis demonstrates that in clinically stable patients without resistant or skin/soft tissue infections, the use of a glycopeptide can be delayed for another 3-4 days. (3) The choice of drugs for monotherapy is currently being evaluated; preliminary results demonstrate that ceftazidime has a significantly inferior response rate (without modification) to other evaluated antibiotics. In conclusion, guidelines should be based on the systematic evaluation of all relevant clinical trials. The analysis of the existing data leads to the recommendation of monotherapy, without aminoglycoside, using piperacillin-tazobactam, cefepime, meropenem or imipenem-cilastin, any of which may be continued for up to 7 days in persistently febrile, clinically stable patients without skin/soft tissue infections. The choice of drug as standard first-line therapy should depend on drug costs, local resistance rates and the potential for resistance induction.

Anti-Bacterial Agents↗

PubMed2005

Contribution of clonal dissemination and selection of mutants during therapy to Pseudomonas aeruginosa antimicrobial resistance in an intensive care unit setting.

Rates of antibiotic resistance in Pseudomonas aeruginosa isolates from intensive care unit (ICU) patients are expected to be dependent on the selection of resistance mutations during therapy, the availability of exogenous resistance determinants and their dissemination potential, and the efficiency of transmission of the resistant strains. The relative contributions of these three factors were studied in an ICU with no apparent outbreak in 216 sequential P. aeruginosa isolates recovered from 102 patients between September 2002 and November 2003. Analysis of pulsed-field gel electrophoresis patterns revealed the presence of 82 different clones. Thus, the dissemination of particular resistant clones had a minimal effect on the relatively high overall resistance frequencies found for imipenem (32%), cefepime (25%), ceftazidime (24%), meropenem (22%), ciprofloxacin (18%) and tobramycin (2%). Rates of primary resistance were relatively low, and resistance development during treatment (secondary resistance) was the main factor contributing to the overall high resistance rates. In ICU settings with a low prevalence of epidemic resistant strains, the main strategy for resistance control should focus on the design of targeted regimens to avoid the development of resistance.

Anti-Bacterial Agents↗

PubMed1994

Isolation and partial purification of a carbapenem-hydrolysing metallo-beta-lactamase from Pseudomonas cepacia.

A metallo-beta-lactamase has been isolated from a clinical strain of Pseudomonas cepacia and partially purified using Cibacron blue F3GA coupled agarose. The resulting preparation showed a single band of beta-lactamase activity (pI 8.45) after analytical isoelectric focusing. The enzyme was particularly effective in the hydrolysis of imipenem. Meropenem, biapenem, cephaloridine, ceftazidime, benzylpenicillin, ampicillin and carbenicillin were also hydrolysed, although at a lower rate. An unusual inhibition profile was noted. Inhibition by the metal ion chelators ethylenediaminetetraacetic acid and o-phenanthroline was reversed by addition of zinc, indicating a metallo-enzyme, whilst > 90% inhibition was attainable with 0.1 mM concentrations of tazobactam and clavulanic acid. A study of 8 other clinical isolates showed the enzyme to be present and inducible by imipenem in each case. This enzyme was assigned PCM-I (Pseudomonas cepacia metalloenzyme I).

Bacterial Proteins↗

PubMed1996

A kinetic study of NMC-A beta-lactamase, an Ambler class A carbapenemase also hydrolyzing cephamycins.

In order to analyze its kinetic parameters, an Ambler class A carbapenemase NMC-A was purified. NMC-A demonstrated unusually strong hydrolytic activity towards imipenem and meropenem. Moreover, it hydrolyzed cephamycins with kcat values uncommonly high for this class of beta-lactamases. Clavulanic acid and tazobactam had comparable inhibitory activity against NMC-A, whereas sulbactam was the least active inhibitor. Noticeably, NMC-A was more readily inhibited by brobactam. All these catalytic properties suggest that NMC-A possesses an original structure of its active site allowing hydrolysis of beta-lactams usually stable to the hydrolytic activity of class A beta-lactamases.

Bacterial Proteins↗

PubMed2005

Antibiotic susceptibility of Listeria monocytogenes in Denmark 1958-2001.

In order to see whether the susceptibility of Danish Listeria monocytogenes strains has changed over the years we examined a collection of human isolates from the period 1958-2001. We, furthermore, wanted to compare L. monocytogenes susceptibility testing using a disc diffusion assay with MIC measurements performed by the E-test. 106 strains isolated predominantly from blood cultures and cerebrospinal fluids were examined together with three reference strains. Susceptibility to the following antibiotics was tested by the E-test and by Oxoid discs using Iso-sensitest agar: penicillin G, ampicillin, meropenem, gentamicin, sulphamethoxazole, trimethoprim, ciprofloxacin, erythromycin, vancomycin, linezolid, chloramphenicol and tetracycline. The strains were in the main sensitive to all antibiotics examined using both methods, except for ciprofloxacin, where the strains were intermediate sensitive. However, for penicillin, ampicillin and sulphamethoxazole, while the disc diffusion assay found the strains to be sensitive, MIC measurements generally placed the strains one dilution above the breakpoint for sensitivity in the intermediate sensitive group. Based on the MIC measurements, the antibiotic susceptibility of L. monocytogenes has not changed in Denmark from 1958 to 2001, and the multiresistant strains found in human infections elsewhere have not been found in Denmark.

Biological Evolution↗

PubMed2005

Dysgonomonas capnocytophagoides bacteraemia in a neutropenic patient treated for acute myeloid leukaemia.

Dysgonomonas capnocytophagoides, formerly known as CDC group DF-3, is an opportunistic pathogen associated with diarrhoea and very rarely bacteraemia. We report a case of D. capnocytophagoides found in blood cultures from a severely neutropenic patient treated for acute myeloid leukaemia. The isolate was found resistant to penicillin, cephalosporins, meropenem, aminoglycosides and ciprofloxacin, and susceptible to ampicillin, tetracycline, chloramphenicol, clindamycin and trimethoprim-sulphamethoxazole. It was identified using conventional phenotypic testing but remained unidentified by the automated identification system (Vitek-2) as this system did not contain DF-3 or D. capnocytophagoides in its database.

Acute Disease↗

PubMed1994

Bacterial biofilm in chronic airway infection.

We hypothesized that bacterial biofilm formation could be an important factor that makes some infections intractable, and conducted the following study to confirm the role of bacterial biofilm in airway infection. We first microscopically examined airway surface in patients with an intractable airway infection and detected bacterial biofilms adhering to the airway surface. Most of the airway biofilm diseases were diffuse panbronchiolitis and bronchiectasia due to Pseudomonas aeruginosa or Klebsiella pneumoniae. In vivo examination revealed that chemotaxis of neutrophils in patients with biofilm bacteria was less than that in patients with floating type bacteria. Interaction of Pseudomonas biofilm with antibacterial agents was examined in vitro. The rate of survival of biofilm bacteria was higher than that of floating bacteria in contact with twice the minimal bactericidal concentration of ciprofloxacin (CPFX) or two to 10 times the minimal inhibitory concentration of cefclidin and meropenem which are highly potent antibacterial agents against P. aeruginosa. Additionally, the effects of clarithromycin (CAM) on biofilm bacteria were studied in order to investigate new therapeutic maneuvers against a bacterial biofilm. Also, the combination of CPFX and CAM was more effective in decreasing the bacterial survival rate than CPFX alone. The results suggest that administration CAM can be one of the therapeutic maneuvers against biofilm bacteria.

Animals↗

PubMed1998

[Early manifestation of septic shock and disseminated intravascular coagulation complicated by acute myocardial infarction in a patient suspected of having Legionnaires' disease].

Legionellosis is an important cause of severe pneumonia in the community. Inadequate therapy will lead to respiratory distress syndrome, disseminated intravascular coagulation (DIC) and finally fatal multiple organ failure. We encountered a rare case in which early manifestation included septic shock and DIC complicated by acute myocardial infarction (AMI) suspected to be derived from Legionnaires' disease. A 54-year-old healthy female complained of lumbago, high fever and dry cough 10 days after visiting a hot spring spa. She was emmergently admitted due to shock. Physical examination demonstrated hypotension, high fever, course creakle in the right lower lung. Hepatosplenomegaly, lymphadenopathy and eruption were not found. WBC count was 34600/microliters with nuclear shift. CRP elevated. FDP, D dimer and TAT also elevated CPK elevated with dominance of the MB isozyme. Chest roentogenography revealed congestive heart failure, pleural effusion and obscure pneumonic shadow and EKG showed ST segment elevation in leads I, II, III, aVF, V4, V5, and V6. The patient was diagnosed as having septic shock, DIC and AMI. She was treated with gabexate mesilate, high dose methyl prednisolone and dopamine hydrochloride as well as piperacillin, meropenem, isepamycin and fluconzaole. Despite intensive care, the blood pressure fell again and pneumonia had progressed on the 8th hospital day. These antibiotics appeared to be ineffective. Erythromycin was then administered and a dramatic effect. was obtained as the patient recovered. Serum titer of Legionella pneumophila (serogroup 1) rose to 128-fold 2 weeks after the onset. Other serum titers such as Chlamydia psittaci, Rickettsia, Mycoplasma were all negative. Cultures obtained from the sputum, throat swab, urine and blood did not yield any microorganisms. Although the diagnosis could not be confirmed because the titer did not elevate over 256-fold of 4-fold within 2 weeks after the onset, Legionella infection was highly suspected from the clinical features. This is a rare case in which septic shock and DIC with AMI preceded pulmonary symptoms in a non-immunocompromised patient.

Diagnosis, Differential↗

PubMed1999

[A case of endophthalmitis and abscesses in the liver and the lung caused by Klebsiella pneumoniae].

[Case report] A 65-year-old male war admitted to a local hospital because of fever. He was treated with piperacillin and clindamycin without noticeable effect. He began to complain of loss of vision on the third hospital day and culture of the blood specimen yielded Klebsiella pneumoniae. He was diagnosed as endophthalmitis and referred to our hospital for further examination. The hematological laboratory test showed leukocytosis (12,700/microliter) and increased CRP (20.4 mg/dl). A computed tomographic (CT) scan of the thorax revealed multiple lung abscesses. An abdominal ultrasonographic scan and a CT scan of the abdomen revealed multiple liver abscesses. We drained the abscess in the liver and Klebsiella pneumoniae was detected from the sample of aspirated fluid and his sputum. Meropenem was administered intravenously. Fever started to improve on the tenth hospital day and the size of both liver and lung abscesses were reduced. He has lost vision of his right eye. He was discharged after sixty days. He did not have any immunosuppressive underlying disease including HIV infection and diabetes mellitus which cause these lesions.

Aged↗

PubMed2001

[Antimicrobial activities and mechanisms of carbapenem resistance in clinical isolates of carbapenem-resistant Pseudomonas aeruginosa and Acinetobacter spp].

We tested the antimicrobial activities of meropenem (MEPM), imipenem (IPM), panipenem (PAPM), piperacillin (PIPC), cefepime (CFPM), aztreonam (AZT), amikacin (AMK), and levofloxacin (LVFX) against 106 clinical Pseudomonas aeruginosa isolates and 64 clinical Acinetobacter spp. isolates with reduced susceptibility to carbapenems. Using NCCLS breakpoints, the percentages of P. aeruginosa strains susceptible to AMK and Acinetobacter spp. strains susceptible to LVFX were found to be 51.1% and 55.6%, respectively, which represented the highest activity among 8 antimicrobial agents in each organism. Referring to the correlations among MICs of carbapenems, MEPM showed a higher activity than IPM and PAPM in both organisms; 29 of the 94 strains (30.9%) of IPM-resistant P. aeruginosa were susceptible to MEPM. Further study for resistance mechanisms to carbapenems by the disk diffusion method using 2-mercaptopropionic acid revealed that 8 of the 64 Acinetobacter spp. isolates (12.5%) were metallo-beta-lactamase producers, while none of 106 P. aeruginosa isolates were metallo-beta-lactamase producers. PCR analysis using blaIMP-specific primers confirmed that 4 of the 8 metallo-beta-lactamase-producing Acinetobacter spp. isolates detected by the disk diffusion method were carrying the blaIMP gene. The identification of metallo-beta-lactamase-producing Acinetobacter spp. isolates implies that metallo-beta-lactamase genes have been disseminated among various gram-negative pathogens.

Acinetobacter↗

PubMed2002

[Resistance to antimicrobiotics and mutation of PBPs in Streptococcus pneumoniae isolated from Kinki region of Japan].

We studied the susceptibility to penicillin G (PCG) and other antimicrobiotics in 235 clinical isolates of Streptococcus pneumoniae. Samples were collected between April 1 and June 30, 2000 from nine medical institutions of the Kinki Region of Japan. We classified the minimum inhibitory concentration (MIC) of PCG according to the National Committee for Clinical Laboratory Standards (NCCLS) criteria. The overall rate of all types of S. pneumoniae resistance was 53.2% (penicillin-susceptible S. pneumoniae (PSSP): 46.8%, penicillin-intermediate S. pneumoniae (PISP): 42.6%, penicillin-resistant S. pneumoniae (PRSP): 10.6%). In other antimicrobiotics, the resistance (R)/intermediate susceptibility (I) rates (R/I%) were as follows: ceftriaxone, 28.9%; cefotaxime, 7.7%; imipenem, 8.9%; meropenem, 9.8%; clarithromycin, 82.6%; clindamycin, 42.1%; levofloxacin, 0.4%; vancomycin, 0%. We used the polymerase chain reaction to study the mutations of the penicillin-binding proteins pbp1 a, pbp2b, and pbp2x in 140 strains of S. pneumoniae in the MIC for PCG was < 0.5 microgram/ml. Among the 109 strains of PSSP, 32 (29.4%) had no mutation and 77 (70.6%) showed mutation of more than one of the pbp mutations. Among the 31 strains of PISP, only 1 strain (3.2%) was not mutated. Since 70.6% of the strains classified as PSSP had pbp mutations, S. pneumoniae clearly can acquire resistance to anti-microbiotics. In the future, a comprehensive surveillance of S. pneumoniae is necessary.

Aminoacyltransferases↗

PubMed2002

[Mutation of penicillin-binding protein genes and antimicrobial susceptibility of Haemophilus influenzae isolated from spinal fluid or blood in children].

Between September 1999 and August 2001, we studied serotypes to capsular antigen, beta-lactamase production, mutation of penicillin binding protein (PBP) genes by PCR method, and antimicrobial susceptibilities of 13 strains of Haemophilus influenzae isolated from spinal fluid or blood in children. Diseases of patients were meningitis in 11, pneumonia in 1, and laryngitis in 1. The age range of the patients was from 26 days to 5 years. The serotypes of all strains were b. Four of the 13 strains were beta-lactamase-positive. The mutation of genes of pbp3 was revealed from 4 isolates and 2 of the strains were beta-lactamase-positive. MICs of ampicillin to beta-lactamase-negative strains ranged from 0.125 to 1 microgram/ml and those to beta-lactamase-positive were more than 32 micrograms/ml. MICs of 2 strains of beta-lactamase-negative and mutation-positive were 0.5 and 1 microgram/ml. The excellent active antimicrobials in our study was cefotaxime (MIC90 0.06 microgram/ml), meropenem (MIC90 0.125 microgram/ml), ceftazidime (MIC90 0.25 microgram/ml), and cefepime (MIC90 0.25 microgram/ml).

Anti-Bacterial Agents↗

PubMed2004

[Molecular epidemiology of Haemophilus influenzae strains isolated from patients with meningitis during 1999 to 2003].

A total of 535 Haemophilus influenzae strains from 226 Japanese institutions participating in the Nationwide Surveillance Study Group for Bacterial Meningitis were sent to our laboratory during 1999 to 2003. All strains were analyzed by PCR to identify the beta-lactam resistance genes, and their susceptibilities to beta-lactam agents were determined. These strains were classified into 6 genotype patterns and MIC90 values for ampicillin (ABPC): (i) beta-lactamase nonproducing, ABPC susceptible (BLNAS) strains and lacked all resistance genes (27.7% of isolates; MIC90, 0.5 microg/ml); (ii) beta-lactamase producing, ABPC resistant (BLPAR) strains (12.9%, 16 microg/ml); (iii) beta-lactamase nonproducing, ABPC resistant (Low-BLNAR) strains with a Asn526Lys amino acid substitution in ftsI gene encoding PBP3 (31.2%, 2 microg/ml); (iv) beta-lactamase nonproducing, ABPC resistant (BLNAR) strains with Ser385Thr and Asn526Lys substitutions in ftsI (17.2%, 8 microg/ml); (v) amoxicillin/clavlanic acid resistant (BLPACR I) strains, having beta-lactamase gene and a Asn526Lys amino acid substitution in ftsI (9.2%, 32 microg/ml); and (vi) amoxicillin/clavlanic acid resistant (BLPACR II), having beta-lactamase gene and ftsI substitutions as for BLNAR strains (1.9%, 64 microg/ml). All but 4 strains were serotype b. The prevalence of BLNAR strains has increased exponentially: 0% (n = 0/41) in 1999, 5.8% (n = 4/69) in 2000, 14.1% (n = 19/139) in 2001, 20.1% (n = 32/159) in 2002, and 29.1% (n = 37/127) in 2003. The MIC90s of BLNAR isolates except for ABPC were as follows: piperacillin, 0.125 microg/ml; ceftriaxone, 0.25 microg/ml; meropenem, 0.5 microg/ml; cefotaxime, 1 microg/ml; panipenem, 2 microg/ml; cefozopran, 16 microg/ml; and cefotiam, 64 microg/ml. To prevent such resistance from increasing, expedited vaccination, correct identification of the BLNAR molecularly, and the proper selection of proper antibiotics based on PK/PD must be taken.

Adult↗

PubMed2005

[Severe community-acquired Acinetobacter baumannii bacteremia with unknown foci].

We report an unusual case of Acinetobacter baumannii (A. baumannii) bacteremia in a man without significant comorbidities. A 50-year-old man who noticed a sudden onset of high fever was admitted to Kobe Nishi City Hospital. After the admission, his condition deteriorated rapidly with development of shock, anuria and agitation. Antibiotic therapy with meropenem and amikacin was begun for suspected septic shock along with the mechanical ventilation and continuous hemodialysis. Venous blood cultures yielded A. baumannii. No definite source of bacteremia was detected. Intensive medical management was effective, and he was discharged on the 27th day. While the most common source of infection that leads to A. baumannii bacteremia is in the respiratory tract, community-acquired bacteremia with unknown foci is considered rare.

Acinetobacter Infections↗

PubMed2005

[Clinical effects of piperacillin and tazobactam/piperacillin on Haemophilus influenzae lower respiratory tract infection in pediatric patients].

OBJECTIVE: The prevalence of beta-lactamase-nonproducing ampicillin-resistant (BLNAR) Haemophilus influenzae (H. influenzae) has been increasing in recent years. Piperacillin (PIPC) is one of a few beta-lactams possessing good activity against BLNAR H. influenzae. We studied clinical efficacy of piperacillin and its beta-lactamase inhibitor, tazobactam/piperacillin (TAZ/PIPC) in children with lower respiratory tract infection caused by H. influenzae including resistance strains. METHODS: Subjects were 20 children with lower respiratory tract infection caused by H. influenzae treated with PIPC 100mg/kg/day (7 cases) or TAZ/PIPC 125mg/kg/day (13 cases). We selected cases from which resistant H. influenzae strains might be detected. Patients received prior antimicrobial therapy within two weeks before admission, or with underlying diseases. We examined patient profiles, clinical efficacy, susceptibilities for 6 beta-lactam antibiotics [PIPC, TAZ/PIPC, ampicillin (ABPC), cefotaxime (CTX), ceftriaxone (CTRX), and meropenem (MEPM)] and analyzed 6 genotype patterns of beta-lactam resistant genes by PCR. RESULTS: Efficacy was 7/7 in patients in PIPC group and 12/13 in patients in TAZ/PIPC group. Diminished efficacy was seen in only one case complicated with severe RSV infection. The susceptibility of all strains but one beta-lactamase producing, ABPC resistant (BLP) strain to PIPC and of all to TAZ/ PIPC was below 0.25 microg/mL. The genotype of the 15 strains isolated from the sputum on administration was as follows; beta-lactamase nonproducing, ABPC-susceptible (gBLNAS) strains were 4, gBLP strain was 1, beta-lactamase nonproducing, and ABPC-resistant (gLow-BLNAR) strains were 2, beta-lactamase nonproducing, ABPC resistant (gBLNAR) strains were 8. CONCLUSION: PIPC and TAZ/PIPC were useful against lower respiratory tract infection caused by H. influenzae including BLNAR in children.

Anti-Bacterial Agents↗

PubMed2006

[Extended-spectrum-beta-lactamase-producing Proteus mirabilis: laboratory-based surveillance in cooperation with 12 clinical laboratories in the Kinki Region of Japan].

We studied 247 strains of Proteus mirabilis collected during the 6 months from November 2003 to April 2004 from 12 clinical laboratories in the Kinki region of Japan for the production of extended-spectrum beta-lactamase (ESBL). Eighteen strains (7.3%) showed MICs for cefpodoxime of > or = 2 microg/mL and 13 strains (5.2%) were positive for the double-disk synergy test. Susceptibility depended on genotype. MICs for cefepime, cefozopran, and cefpirome were high (> or = 8 microg/mL), and that for ceftazidime was low (0.12-0.5 microg/mL). Meropenem showed the lowest MIC (< or = 0.03-0.25 microg/mL) of the calbapenems, while other calbapenems showed somewhat higher values (0.5-2 microg/mL). The MIC of tazobactam/piperacillin was also relatively low (< or = 0.25-1 microg/mL). Analysis of the ESBL genotype by the polymerase chain reaction showed that 12 of 13 strains were CTX-M2 types. CTX-M9 was detected in a single laboratory. The clinical background showed 5 strains in urine samples. Twelve of 13 strains were detected in patients with minimal devices use. No symptoms were found in most cases of established syndrome. Analysis of PCR fingerprint profiles of random amplified polymorphic DNA patterns showed that 6 of 7 strains from hospital 1 showed the same pattern, and 5 of 5 strains from hospital 13 showed the same pattern, suggesting the nosocomial spread of P. mirabilis in each hospital.

Drug Resistance, Bacterial↗