cefoxitin and Pseudomonas-Infections

Pseudomonas aeruginosa possessing chromosomally inducible blaPDCalong with other intrinsic mechanism causes infection with high mortality rate. It is difficult to detect inducible AmpC enzymes in this organism and is usually overlooked by routine testing that may lead to therapeutic failure. Therefore, three different inducers were evaluated in the present study to assess their ability of induction of blaPDCin P. aeruginosa.. A total of 189 consecutive Pseudomonas isolates recovered from different clinical specimens (November 2011-April 2013) were selected for the study. Isolates were screened with cefoxitin for AmpC β-lactamases and confirmed by modified three-dimensional extract test (M3DET). Inductions were checked using three inducers, namely, clavulanic acid, cefoxitin and imipenem along with ceftazidime. Molecular screening of AmpC β-lactamase genes was performed by PCR assay. Antimicrobial susceptibility and minimum inhibitory concentrations (MICs) were determined, and repetitive extragenic palindromic-PCR of all blaPDCharbouring isolates was performed.. Inducible phenotype was observed in 42 (24.3%) of 97 (56%) isolates confirmed by M3DET. Among these, 22 isolates harboured chromosomal blaPDCgene, and cocarriage of both chromosomal and plasmid-mediated blaAmpC genes was observed in seven isolates. Cefoxitin-ceftazidime-based test gave good sensitivity and specificity for detecting inducible AmpC enzymes. Isolates harbouring blaPDCshowed high MIC against all tested cephalosporins and monobactam. DNA fingerprinting of these isolates showed 22 different clones of P. aeruginosa.. P. aeruginosa harbouring inducible (chromosomal) and plasmid-mediated AmpC β-lactamase is a matter of concern as it may limit therapeutic option. Using cefoxitin-ceftazidime-based test is simple and may be used for detecting inducible AmpC β-lactamase amongst P. aeruginosa.

Topics: Bacterial Proteins; beta-Lactamases; Cefoxitin; Cephalosporin Resistance; Cephalosporins; DNA Fingerprinting; Humans; Microbial Sensitivity Tests; Plasmids; Pseudomonas aeruginosa; Pseudomonas Infections

This cross-sectional study, performed in an oncology hospital in Goiania, aimed to characterize the prevalence of oral colonization and antimicrobial susceptibility of Pseudomonas spp. isolated from the saliva of healthcare workers. Microorganisms were subjected to biochemical tests, susceptibility profile, and phenotypic detection. Of 76 participants colonized with Gram negative bacilli, 12 (15.8%) harbored Pseudomonas spp. Of all isolates, P. aeruginosa (75.0%), P. stutzeri (16.7%), and P. fluorescens (8.3%), were resistant to cefoxitin, and therefore likely to be AmpC producers. The results are clinically relevant and emphasize the importance of surveillance to minimize bacterial dissemination and multiresistance.

Topics: Anti-Bacterial Agents; beta-Lactam Resistance; Brazil; Cancer Care Facilities; Cefoxitin; Cross-Sectional Studies; Health Personnel; Humans; Microbial Sensitivity Tests; Mouth; Prevalence; Pseudomonas; Pseudomonas Infections; Saliva

Pseudomonas luteola (formerly classified as CDC group Ve-1 and named Chryseomonas luteola) is an unusual pathogen implicated in rare but serious infections in humans. A novel beta-lactamase gene, bla(LUT-1), was cloned from the whole-cell DNA of the P. luteola clinical isolate LAM, which had a weak narrow-spectrum beta-lactam-resistant phenotype, and expressed in Escherichia coli. This gene encoded LUT-1, a 296-amino-acid Ambler class A beta-lactamase with a pI of 6 and a theoretical molecular mass of 28.9 kDa. The catalytic efficiency of this enzyme was higher for cephalothin, cefuroxime, and cefotaxime than for penicillins. It was found to be 49% to 59% identical to other Ambler class A beta-lactamases from Burkholderia sp. (PenA to PenL), Ralstonia eutropha (REUT), Citrobacter sedlakii (SED-1), Serratia fonticola (FONA and SFC-1), Klebsiella sp. (KPC and OXY), and CTX-M extended-spectrum beta-lactamases. No gene homologous to the regulatory ampR genes of class A beta-lactamases was found in the vicinity of the bla(LUT-1) gene. The entire bla(LUT-1) coding region was amplified by PCR and sequenced in five other genetically unrelated P. luteola strains (including the P. luteola type strain). A new variant of bla(LUT-1) was found for each strain. These genes (named bla(LUT-2) to bla(LUT-6)) had nucleotide sequences 98.1 to 99.5% identical to that of bla(LUT-1) and differing from this gene by two to four nonsynonymous single nucleotide polymorphisms. The bla(LUT) gene was located on a 700- to 800-kb chromosomal I-CeuI fragment, the precise size of this fragment depending on the P. luteola strain.

Topics: Amino Acid Sequence; Anti-Bacterial Agents; beta-Lactamases; Catheters, Indwelling; Chromosomes, Bacterial; Cloning, Molecular; Electrophoresis, Gel, Pulsed-Field; Genes, Bacterial; Genetic Variation; Isoelectric Focusing; Kinetics; Microbial Sensitivity Tests; Molecular Sequence Data; Phylogeny; Plasmids; Pseudomonas; Pseudomonas Infections; Recombinant Proteins; Reverse Transcriptase Polymerase Chain Reaction; Substrate Specificity

A Pseudomonas aeruginosa isolate recovered in Belgium produced a novel extended-spectrum ss-lactamase, BEL-2, differing from BEL-1 by a single Leu162Phe substitution. That modification significantly altered the kinetic properties of the enzyme, increasing its affinity for expanded-spectrum cephalosporins. The bla(BEL-2) gene was identified from a P. aeruginosa isolate clonally related to another bla(BEL-1)-positive isolate.

Topics: Belgium; beta-Lactamases; Cephalosporinase; Cephalosporins; Cross Infection; Escherichia coli; Genes, Bacterial; Humans; Kinetics; Microbial Sensitivity Tests; Pseudomonas aeruginosa; Pseudomonas Infections

GES-13 beta-lactamase, a novel GES variant possessing Lys-104 and Asn-170, was identified in Pseudomonas aeruginosa. bla(GES-13) was the single gene cassette of a class 1 integron probably located in the chromosome. GES-13 efficiently hydrolyzed broad-spectrum cephalosporins and aztreonam. Imipenem was a potent inhibitor of GES-13 but was not hydrolyzed at measurable rates.

Topics: Anti-Bacterial Agents; Aztreonam; beta-Lactamases; beta-Lactams; Cephalosporins; Genetic Variation; Humans; Integrons; Microbial Sensitivity Tests; Molecular Sequence Data; Pseudomonas aeruginosa; Pseudomonas Infections; Sequence Analysis, DNA

CXA-101, previously designated FR264205, is a new antipseudomonal cephalosporin. We evaluated the activity of CXA-101 against a highly challenging collection of beta-lactam-resistant Pseudomonas aeruginosa mutants selected in vitro and after antipseudomonal treatment of intensive care unit (ICU) patients. The in vitro mutants investigated included strains with multiple combinations of mutations leading to several degrees of AmpC overexpression (ampD, ampDh2, ampDh3, and dacB [PBP4]) and porin loss (oprD). CXA-101 remained active against even the AmpD-PBP4 double mutant (MIC = 2 microg/ml), which shows extremely high levels of AmpC expression. Indeed, this mutant showed high-level resistance to all tested beta-lactams, except carbapenems, including piperacillin-tazobactam (PTZ), aztreonam (ATM), ceftazidime (CAZ), and cefepime (FEP), a cephalosporin considered to be relatively stable against hydrolysis by AmpC. Moreover, CXA-101 was the only beta-lactam tested (including the carbapenems imipenem [IMP] and meropenem [MER]) that remained fully active against the OprD-AmpD and OprD-PBP4 double mutants (MIC = 0.5 microg/ml). Additionally, we tested a collection of 50 sequential isolates that were susceptible or resistant to penicillicins, cephalosporins, carbapenems, or fluoroquinolones that emerged during treatment of ICU patients. All of the mutants resistant to CAZ, FEP, PTZ, IMP, MER, or ciprofloxacin showed relatively low CXA-101 MICs (range, 0.12 to 4 microg/ml; mean, 1 to 2 microg/ml). CXA-101 MICs of pan-beta-lactam-resistant strains ranged from 1 to 4 microg/ml (mean, 2.5 microg/ml). As described for the in vitro mutants, CXA-101 retained activity against the natural AmpD-PBP4 double mutants, even when these exhibited additional overexpression of the MexAB-OprM efflux pump. Therefore, clinical trials are needed to evaluate the usefulness of CXA-101 for the treatment of P. aeruginosa nosocomial infections, particularly those caused by multidrug-resistant isolates that emerge during antipseudomonal treatments.

Topics: Anti-Bacterial Agents; Bacterial Proteins; beta-Lactam Resistance; beta-Lactamases; Cephalosporins; Cross Infection; Humans; Intensive Care Units; Microbial Sensitivity Tests; Mutation; Pseudomonas aeruginosa; Pseudomonas Infections

A carbapenem-resistant Pseudomonas stutzeri strain isolated from a Dutch patient was analyzed in detail. This isolate produced a metallo-beta-lactamase (MBL) whose gene, with 43.5% GC content, was cloned and expressed in Escherichia coli. beta-Lactamase DIM-1 (for Dutch imipenemase) was weakly related to other Ambler class B beta-lactamases, sharing <52% amino acid identity with the most closely related MBL, GIM-1, and 45% identity with IMP-type MBLs. The beta-Lactamase DIM-1 significantly hydrolyzed broad-spectrum cephalosporins and carbapenems and spared aztreonam. This MBL gene was embedded in a class 1 integron containing two other gene cassettes, encoding resistance to aminoglycosides and disinfectants, that was located on a 70-kb plasmid.

Topics: Amino Acid Sequence; Bacterial Proteins; Base Sequence; beta-Lactamases; Cloning, Molecular; DNA, Bacterial; Drug Resistance, Bacterial; Genes, Bacterial; Humans; Integrons; Middle Aged; Molecular Sequence Data; Netherlands; Pseudomonas Infections; Pseudomonas stutzeri; Sequence Homology, Amino Acid; Substrate Specificity

Genetic structures surrounding the carbapenem-hydrolyzing Ambler class A bla KPC gene were characterized in several KPC-positive Klebsiella pneumoniae and Pseudomonas aeruginosa strains isolated from the United States, Colombia, and Greece. The bla KPC genes were associated in all cases with transposon-related structures. In the K. pneumoniae YC isolate from the United States, the beta-lactamase bla KPC-2 gene was located on a novel Tn3-based transposon, Tn4401. Tn4401 was 10 kb in size, was delimited by two 39-bp imperfect inverted repeat sequences, and harbored, in addition to the beta-lactamase bla KPC-2 gene, a transposase gene, a resolvase gene, and two novel insertion sequences, ISKpn6 and ISKpn7. Tn4401 has been identified in all isolates. However, two isoforms of this transposon were found: Tn4401a was found in K. pneumoniae YC and K. pneumoniae GR from the United States and Greece, respectively, and differed by a 100-bp deletion, located just upstream of the bla KPC-2 gene, compared to the sequence of Tn4401b, which was found in the Colombian isolates. In all isolates tested, Tn4401 was flanked by a 5-bp target site duplication, the signature of a recent transposition event, and was inserted in different open reading frames located on plasmids that varied in size and nature. Tn4401 is likely at the origin of carbapenem-hydrolyzing beta-lactamase KPC mobilization to plasmids and its further insertion into various-sized plasmids identified in nonclonally related K. pneumoniae and P. aeruginosa isolates.

Topics: Base Sequence; beta-Lactamases; beta-Lactams; Cloning, Molecular; Colombia; DNA Transposable Elements; Greece; Humans; Klebsiella Infections; Klebsiella pneumoniae; Microbial Sensitivity Tests; Molecular Sequence Data; Pseudomonas aeruginosa; Pseudomonas Infections; Sequence Analysis, DNA; United States

A detailed biochemical characterization of the Pseudomonas aeruginosa VIM-11 metallo-beta-lactamase (MbetaL) is reported. The only substitution differentiating VIM-11 from VIM-2 (N165S) promoted a slightly improved catalytic efficiency of the former on 3 out of 12 substrates, notably the bulky cephalosporins. Thus, MbetaL-mediated resistance also may be modulated by remote mutations.

Topics: beta-Lactam Resistance; beta-Lactamases; beta-Lactams; Binding Sites; Escherichia coli; Humans; Kinetics; Microbial Sensitivity Tests; Models, Molecular; Plasmids; Pseudomonas aeruginosa; Pseudomonas Infections

Topics: Abdominal Abscess; Appendicitis; Cefoxitin; Ceftriaxone; Cefuroxime; Child; Community-Acquired Infections; Drug Resistance, Microbial; Enterobacteriaceae; Enterobacteriaceae Infections; Humans; Metronidazole; Pseudomonas; Pseudomonas Infections; Randomized Controlled Trials as Topic

Most of the open pelvic fractures are associated with skin wounds of varying anatomy. Injuries of the pelvis, which only momentarily communicate with the exterior or into a hollow organ are often difficult to diagnose. We managed a patient with an occult open displaced fracture of the public rami that had probably torn the hip capsule and was not recognised on admission as the open wound was in the vaginal wall. It subsequently resulted in septic arthritis of the ipsilateral hip joint. Vaginal and rectal examination in grossly displaced pelvic fractures should be done to diagnose at an early stage any occult open wound which may pose high risk of contamination of the fracture site.

Topics: Adult; Amikacin; Arthritis, Infectious; Cefoxitin; Drug Therapy, Combination; Female; Fractures, Open; Hip Joint; Humans; Klebsiella Infections; Pelvic Bones; Pseudomonas Infections

A study of determinants of outcome in adult patients with intra-abdominal or skin/soft tissue infections treated with cefotetan, cefoxitin, or ampicillin/sulbactam monotherapy was undertaken. Patients were matched for principal infectious process, surgery performed for the management of the infection, year of hospital admission, age, and sex. The criteria for inclusion, exclusion, and matching of patients and assignment of clinical and microbiological outcome were based on the 1992 Infectious Diseases Society of America/Federal Drug Administration guidelines for the evaluation of anti-infective drug products. One hundred and thirty-seven cases of intra-abdominal or skin and soft tissue infections treated with cefotetan (n = 47), cefoxitin (n = 43), or ampicillin/sulbactam (n = 47) monotherapy were selected without knowledge of outcome and analyzed using a single blinded analysis. The baseline characteristics did not differ between the treatment groups, nor did the rates of clinical or microbiological failure. A multivariate analysis showed that isolation of an organism resistant to the treatment regimen, including Pseudomonas spp., [odds ratio (OR) = 14.9, p = 0.001], being on antibiotic therapy at the time of admission (OR = 4.5, p = 0.007), and diagnosis of a complicated intra-abdominal infection (OR = 3.5, p = 0.014) were independently associated with clinical failure. These data support the assertion that antibiotic resistant organisms in mixed anaerobic/aerobic infections are associated with clinical failure and suggest that the antibiotic regimen should be modified to include Pseudomonas spp. in its spectrum when this organism is isolated from patients with such infections.

Topics: Abdominal Abscess; Adult; Age Factors; Aged; Ampicillin; Anti-Bacterial Agents; Cefotetan; Cefoxitin; Cephamycins; Drug Resistance, Microbial; Female; Gram-Negative Bacterial Infections; Gram-Positive Bacterial Infections; Humans; Male; Matched-Pair Analysis; Middle Aged; Multivariate Analysis; Penicillins; Pseudomonas Infections; Retrospective Studies; Risk Factors; Sex; Skin Diseases, Bacterial; Soft Tissue Infections; Sulbactam; Treatment Outcome

The efficacy of beta-lactam antibiotics and amikacin alone and in various combinations against Pseudomonas aeruginosa was studied in a rabbit model simulating a closed-space infection in a locally neutropenic site. Six strains of P. aeruginosa were studied in semipermeable chambers placed subcutaneously in rabbits. Therapy was begun 4 h after inoculation of 5 X 10(4) CFU of bacteria per ml of pooled rabbit serum into the chambers. Antibiotics were administered intramuscularly every 6 h for 16 doses. Quantitative bacteriology was measured at the start of therapy and at 20, 44, and 92 h thereafter. Antibiotic concentrations were measured in blood and chamber fluid. Results were compared with in vitro tests of susceptibility and synergy. No single-agent therapy eradicated any of the six test organisms. Azlocillin (100 mg/kg per dose) plus amikacin (20 mg/kg per dose) eliminated five of six organisms by 92 h, and ceftizoxime (100 mg/kg per dose) plus amikacin (20 mg/kg per dose) eliminated three of six test strains. Azlocillin plus ceftizoxime (each 100 mg/kg per dose) failed to eliminate any of the six strains. To eliminate P. aeruginosa in this model, two drugs were required, with one being an aminoglycoside. In vitro susceptibility tests of synergy were predictive of successful therapy whenever the antibiotic concentrations (free and total) at the infection site exceeded the MBC for both the aminoglycoside alone and the beta-lactam when tested in combination with amikacin.

Topics: Agranulocytosis; Amikacin; Animals; Anti-Bacterial Agents; Azlocillin; Cefotaxime; Cefoxitin; Ceftizoxime; Disease Models, Animal; Drug Combinations; Female; Hydrogen-Ion Concentration; Kanamycin; Microbial Sensitivity Tests; Neutropenia; Penicillins; Protein Binding; Pseudomonas Infections; Rabbits

Our clinical experience with new antibiotics giving special consideration to the individual cephalosporin groups is discussed. Although newer cephalosporins from cefamandol and cefoxitin to cefotiam and cefoperazon already showed increased effectiveness (for example, cefoxitin in bacteroides infection) in comparison to older ones, the real breakthrough regarding enterobacteriaceae was only made with cephalosporins of the cefotaxime group. This group's main indication is non-specific initial therapy of severe nosocomial infections, especially processes in which the presence of resistant enterobacteriaceae must be assumed. Because of its broad spectrum of action, cefotaxime can to a large extent replace the combinations with aminoglycosides which were used previously. When required, cefotaxime proves to be a good partner for combinations with pseudomonas antibiotics.

Topics: Bacteroides Infections; Cefotaxime; Cefoxitin; Cephalosporins; Cross Infection; Drug Therapy, Combination; Enterobacteriaceae Infections; Gentamicins; Humans; Pseudomonas Infections; Staphylococcal Infections; Structure-Activity Relationship