I. Boutiba-Ben Boubaker

Nosocomial outbreak of imipenem-resistant Pseudomonas aeruginosa producing VIM-2 metallo-β-lactamase in a kidney transplantation unit

BackgroundTwenty four non replicate imipenem resistant P. aeruginosa were isolated between January and November 2008, in the kidney transplantation unit of Charles Nicolle Hospital of Tunis (Tunisia). This study was conducted in order to establish epidemiological relationship among them and to identify the enzymatic mechanism involved in imipenem resistance.MethodsAnalysis included antimicrobial susceptibility profile, phenotypic (imipenem-EDTA synergy test) and genotypic detection of metallo-β-lactamase (MBL) (PCR), O-serotyping and pulsed-field gel electrophoresis.ResultsAll strains showed a high level of resistance to all antimicrobials tested except to colistin. The presence of MBL showed concordance between phenotypic and genotypic methods. Sixteen isolates were identified as VIM-2 MBL-producers and 13 of them were serotype O4 and belonged to a single pulsotype (A).ConclusionsThis study describes an outbreak of VIM-2-producing P. aeruginosa in a kidney transplantation unit. Clinical spread of blaVIM-2 gene is a matter of great concern for carbapenem resistance in Tunisia.

Emergence of carbapenem-resistant OXA-48 carbapenemase-producing Enterobacteriaceae in Tunisia

We screened 21 extended spectrum β-lactamase-producing Enterobacteriaceae with reduced susceptibility to carbapenems for carbapenemase production. Five strains (four Klebsiella pneumoniae and one Citrobacter freundii) showed carbapenemase production, which was identified as OXA-48. The bla(OXA-48) gene was detected on ~54 kb plasmids belonging to IncA/C in one case. Two isolates harboured IS1999, which is involved in bla(OXA-48) mobilization. Carbapenem resistance in enterobacteria should be regarded as an emerging clinical problem in our hospital and necessitates rigorous surveillance in order to limit its spread.

Macrolide and tetracycline resistance in clinical strains of Streptococcus agalactiae isolated in Tunisia

Between 2007 and 2009, 226 clinical strains of Streptococcus agalactiae, recovered from female genital specimens and from gastric fluid or ear specimens from infected newborns, were isolated at the Laboratory of Microbiology of Charles Nicolle Hospital of Tunis. They were investigated to determine the prevalence of antibiotic resistance and to characterize the mechanisms of resistance to macrolide and tetracycline. All strains were susceptible to penicillin, ampicillin and quinupristin-dalfopristin. They were resistant to chloramphenicol (3.1%), rifampicin (19.1%), erythromycin (40%) and tetracycline (97.3%); 3.1% were highly resistant to streptomycin and 1.3% to gentamicin. Among the erythromycin-resistant isolates, 78.7% showed a constitutive macrolide-lincosamide-streptogramin B (MLS(B)) phenotype with high-level resistance to macrolides and clindamycin (MIC(50) >256 µg ml(-1)), 10% showed an inducible MLS(B) phenotype with high MICs of macrolides (MIC(50) >256 µg ml(-1)) and low MICs of clindamycin (MIC(50)=8 µg ml(-1)) and 2.2% showed an M phenotype with a low erythromycin-resistance level (MIC range=12-32 µg ml(-1)) and low MICs of clindamycin (MIC range: 0.75-1 µg ml(-1)). All strains were susceptible to quinupristin-dalfopristin and linezolid (MIC(90): 0.75 µg ml(-1) for each). MLS(B) phenotypes were genotypically confirmed by the presence of the erm(B) gene and the M phenotype by the mef(A) gene. Resistance to tetracycline was mainly due to the tet(M) gene (93.1%) encoding a ribosome protection mechanism. This determinant is commonly associated with the conjugative transposon Tn916 (P≤0.0002). tet(O) and tet(T) existed in a minority (2.2% and 0.4%, respectively). The efflux mechanism presented by tet(L) was less frequently present (4.5%). No significant association was found between erm(B) and tet(M) genes.

Epidemiological markers of Streptococcus pyogenes strains in Tunisia

To further understand the epidemiology of Streptococcus pyogenes or group A streptococcus (GAS) infections in Tunisia, phenotypic and genomic markers of GAS isolates, including antibiotic susceptibility, biotypes, T and emm types and toxin gene profiles, have been characterized. A total of 103 isolates, collected between 2000 and 2006, were investigated; 47 were recovered from invasive infections, and 56 from non-invasive infections. Rates of resistance to tetracycline, erythromycin, clindamycin and rifampin were 70.8%, 4.8%, 4.8% and 0.9%, respectively. High levels of resistance to streptomycin and kanamycin were observed in 1.9% and 4.8% of isolates, respectively. Biotype 3 was most common. Twenty different T patterns were observed, with a predominance of T3/13/B3264, and 38 different emm types. In both invasive and non-invasive isolates, emm118 (9.7%), emm42 (8.7%), emm1 (7.8%), st432 (6.8%), emm28 (5.8%) and emm76 (5.8%) were the most prevalent types; emm1, emm76 and emm18 were mainly observed among invasive infections, whereas emm118 (12.5%), emm42 (10.7%) and emm28 (8.9%) were predominant among non-invasive infections. The speB gene was detected in all isolates, but there were variable frequencies of speA, speC and ssa (20.3%, 32% and 25.2% respectively). Significant associations of emm1, emm18 and emm3 with speA and of emm4 and st432 with ssa were found. This first report from Tunisia revealed a unique emm distribution of GAS that differs from those of other regions. This information on the distribution of such emm types will be useful for the development of an appropriate vaccine in a country where the incidence of rheumatic fever remains high.

Virulence determinants, biofilm production and antimicrobial susceptibility in Staphylococcus aureus causing device-associated infections in a Tunisian hospital

Staphylococcus aureus is a clinically relevant pathogen that causes device-related infections (DRI) driven by several virulence factors. This study characterized S. aureus isolates involved in DRI in Tunisian patients. Forty consecutive S. aureus strains causing DRI and 47 randomly selected S. aureus strains causing non-device-related infections (NDRI) were collected. All strains were screened phenotypically for antibiotic susceptibility and biofilm forming ability. They were investigated for accessory gene regulator (agr) types, biofilm encoding genes (icaADBC), adhesins, leukotoxins, toxic shock toxin, enterotoxins and exotoxins encoding genes by polymerase chain reaction. Meticillin-resistant S. aureus (MRSA) strains were further characterized by staphylococcal cassette chromosome mec (SCCmec) typing. MRSA rates among DRI and NDRI isolates were 23% and 49% (P=0.02), respectively. The DRI isolates formed biofilm more frequently (n=32) than the NDRI isolates (n=28) (P=0.04), with predominance of the moderate biofilm producer category (P=0.027). All biofilm-positive isolates except four harboured icaADBC genes. A significant difference was observed between DRI and NDRI isolates for fnbA (53-77%), spa (45-26%), sdrD (80-55%) and sen (33-11%) genes. DRI strains were agrI (48%) and agrII (30%) types, whereas NDRI strains were agrI (36%) and agrIII (43%) types. SCCmec type IV was carried by 50% of MRSA isolates. This study highlights the virulence potential displayed by S. aureus isolated from DRI in comparison with NDRI.