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Acinetobacter baumannii and Pseudomonas aeruginosa are among the most prevalent pathogens causing a wide range of serious infections in hospitalized patients and contaminating intensive care units and inanimate surfaces. The purpose of this study was to investigate the mechanism of carbapenem resistance in clinical and hospital environmental isolates of A. baumannii and P. aeruginosa recovered from a Libyan hospital. From a total of 82 Gram-negative bacteria, 8 isolates of A. baumannii and 3 isolates of P. aeruginosa exhibited resistance to imipenem with minimum inhibitory concentrations ranging from 16 to >32 μg/mL. Five isolates of A. baumannii harbored bla OXA-23 gene, from which three isolates were collected from patients and two from hospital environment. Only one isolate harbored bla NDM-1 gene, which was responsible for carbapenem resistance in A. baumannii . The OprD gene seems to be disturbed by an insertion sequence ( IS ) in two isolates and affected by polymorphism in one isolate. Pulsed-field gel electrophoresis results showed high genetic diversity among carbapenemase producing A. baumannii . This study highlights the dissemination of bla OXA-23 and bla NDM-1 genes in a Libyan setting. Therefore, infection prevention and control practices, antimicrobial stewardship initiatives, and antimicrobial resistance surveillance systems should be implemented to prevent the wide spread of antimicrobial resistance.

We propose a simple and rapid method to discriminate SHV-type extended spectrum beta-lactamase (ESBL) genes in P. aeruginosa based on PCR techniques (PCR-RFLP and RSI-PCR). We studied 22 producing ESBL P. aeruginosa strains isolated from seven immunocompromised patients (19 isolates) and from environmental swabs (three isolates) at the Bone Marrow Transplantation Center of Tunis. Screening PCR with primer pairs designed to detect gene encoding TEM, SHV, OXA group I, OXA group II, OXA-18 and PER-1 ESBL was positive for bla(OXA18) and bla(SHV) genes in all isolates. Pulsed field gel electrophoresis using SpeI endonuclease defined five genotypic groups. For at least one isolate corresponding to each genotype observed, restriction of PCR products by DdeI and BsrI revealed the same restriction pattern that the bla(SHV-1) negative control; in the same way, RSI-PCR products digestion by NruI, thus excluding 35, 238 and 240 mutations characterizing reported ESBL in P. aeruginosa (SHV-2a, SHV5 et SHV12), and suggesting that studied bla(SHV) genes were not ESBL ones. Genomic DNA hybridization by southern blot with probe consisting in bla(SHV-1) gene was positive in these isolates. Sequencing the full-length open reading frame revealed nucleotide sequence of the bla(SHV-1). PCR-RFLP and RSI-PCR results were then confirmed. This approach is effective for screening P. aeruginosa for ESBL genes carriage in epidemiological studies and for detecting new variants.

Twelve multidrug-resistant Pseudomonas aeruginosa (MDRPA) isolates were recovered over a period of two years in the National Bone Marrow Transplant Centre of Tunisia. MDRPA isolates were isolated from seven patients and from three environmental samples. Isoelectric focusing revealed pIs of 8.2, 5.5 and 7.6 in all MDRPA isolates. These strains produced the OXA-18 extended spectrum beta-lactamase and an SHV type beta-lactamase as shown by screening PCR analysis. DNA hybridization confirmed this inference, detecting bla(SHV) gene in these isolates. Pulsed-field gel electrophoresis (PFGE) defined one predominant genomic group; group A (seven isolates) and four different genotypes containing one to two isolates. Clonally related isolates were recovered from three patients and from two washbasins. Sequencing DNA of cluster representative strains identified the classical bla(SHV-1) gene. For these strains, the nucleotide sequence of the structural bla(SHV-1) gene was nearly identical to those previously described. Such enzyme has not been reported from P. aeruginosa. This is the first report of the SHV-1 penicillinase in epidemic P. aeruginosa strain.

Following systematic screening for ceftazidime-resistant (CAZ-R) Pseudomonas aeruginosa, 24 isolates producing extended-spectrum beta-lactamase (ESBL) were recovered during a 24-month period at the National Bone Marrow Transplant Centre of Tunisia. These isolates were from seven immunocompromised patients and from environmental swabs. ESBLs inhibited by clavulanic acid were detected by double-disk diffusion tests. Isoelectric focusing revealed that these isolates produced two to four beta-lactamases with pIs of 5.5, 6.1, 6.4, 7.6 or 8.2, and PCR detected the presence of bla(OXA-18), bla(SHV) and bla(TEM) genes in 24, 21 and two isolates, respectively. Pulsed-field gel electrophoresis defined two dominant genotypic groups: group A (16 isolates) and group B (four isolates). Sequencing of PCR products from representative isolates identified the bla(OXA-18) gene and revealed nucleotide sequences belonging to the bla(SHV-1) and bla(TEM-1) genes. Isolates producing OXA-18 belonged to genomic group A and were isolated from four immunocompromised patients in the haematology and graft units, and from two wash-basins in the graft unit. No immunocompromised patient harboured the clonal epidemic strain upon admission. This is the first report of the OXA-18-type ESBL in P. aeruginosa in Tunisia, and the first description of an outbreak caused by an OXA-18-producing strain of P. aeruginosa.

Objective: Investigation of an outbreak caused by an imipenem-resistant Pseudomonas aeruginosa strain and research of its hospital reservoir., Patients and Methods: Nine strains isolated from protected tracheal specimens during 14 weeks (October 2004 to January 2005) from 8 infants, and one strain from vacuum interrupter were studied. Epidemiological study was investigated by determination of antibiotics susceptibility, serotyping and Pulsed-field gel electrophoresis (PFGE)., Results: Strains were of O:12 serotype, they have the same antibiotype characterised by imipenem resistance. Strains were indistinguishable or closely related as determined by PFGE. The common source of P. aeruginosa O:12 strains was not determined, however eradication of the epidemic strain was obtained by amelioration of hygiene conditions and the change of disinfectors., Conclusion: Outbreak of respiratory infections due to an imipenem-resistant P. aeruginosa O:12. The common source of the epidemic strain was not determined.

Objective: Our study dealt with antibiotic resistance and serotypes of Pseudomonas aeruginosa strains isolated from immunocompromised patients in the National Bone Marrow Transplant Center of Tunis as well as molecular typing of ceftazidime resistant strains (CAZ-R)., Design: We studied a total of 87 non-replicate P. aeruginosa isolates from 36 patients (84 strains) or the hospital environment (3 strains)., Results: Rates of antimicrobial resistance were 36% for ceftazidime, 16% for imipenem, 38% for amikacin, and 57% for ciprofloxacin. The 31 CAZ-R strains were associated with O:11 serotype in 84% of the cases. Genetically characterization of CAZ-R strains by Pulsed Field Gel Electrophoresis (PFGE) after digestion of genomic DNA with SpeI revealed 2 genotypic groups. The first was composed of strains isolated from one outpatient between November 1998 and April 1999. Resistance phenotypes of these strains varied after use of antimicrobial drugs. The second was predominant (18/31 CAZ-R strains) in both hematology and graft units and persisted from June 1998 to June 2000 among 5/8 patients. These strains had O:11 serotype in 78% of the cases. The strains of this group were not isolated on patient admission and were isolated from 2 washbasins in the graft unit in May 1999., Conclusion: These results suggest the spread of multidrug-resistant O:11 P. aeruginosa clone from a tap water among hospitalized patients in our center, emphasizing the need of standard control of washbasins to eradicate this reservoir.

Following systematic screening for ceftazidime-resistant (CAZ-R) Pseudomonas aeruginosa, 24 isolates producing extended-spectrum β-lactamase (ESBL) were recovered during a 24-month period at the National Bone Marrow Transplant Centre of Tunisia. These isolates were from seven immunocompromised patients and from environmental swabs. ESBLs inhibited by clavulanic acid were detected by double-disk diffusion tests. Isoelectric focusing revealed that these isolates produced two to four β-lactamases with pIs of 5.5, 6.1, 6.4, 7.6 or 8.2, and PCR detected the presence of blaOXA-18, blaSHV and blaTEM genes in 24, 21 and two isolates, respectively. Pulsed-field gel electrophoresis defined two dominant genotypic groups: group A (16 isolates) and group B (four isolates). Sequencing of PCR products from representative isolates identified the blaOXA-18 gene and revealed nucleotide sequences belonging to the blaSHV-1 and blaTEM-1 genes. Isolates producing OXA-18 belonged to genomic group A and were isolated from four immunocompromised patients in the haematology and graft units, and from two wash-basins in the graft unit. No immunocompromised patient harboured the clonal epidemic strain upon admission. This is the first report of the OXA-18-type ESBL in P. aeruginosa in Tunisia, and the first description of an outbreak caused by an OXA-18-producing strain of P. aeruginosa. [ABSTRACT FROM AUTHOR]

Abstract: Objective: Investigation of an outbreak caused by an imipenem-resistant Pseudomonas aeruginosa strain and research of its hospital reservoir. Patients and methods: Nine strains isolated from protected tracheal specimens during 14 weeks (October 2004 to January 2005) from 8 infants, and one strain from vacuum interrupter were studied. Epidemiological study was investigated by determination of antibiotics susceptibility, serotyping and Pulsed-field gel electrophoresis (PFGE). Results: Strains were of O:12 serotype, they have the same antibiotype characterised by imipenem resistance. Strains were indistinguishable or closely related as determined by PFGE. The common source of P. aeruginosa O:12 strains was not determined, however eradication of the epidemic strain was obtained by amelioration of hygiene conditions and the change of disinfectors. Conclusion: Outbreak of respiratory infections due to an imipenem-resistant P. aeruginosa O:12. The common source of the epidemic strain was not determined. [Copyright &y& Elsevier]