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.

Virulence determinants, phylogenetic groups and fluoroquinolone resistance in Escherichia coli isolated from cystitis and pyelonephritis.

AIM OF THE STUDY: The aim of this study is to assess the relation between virulence genotype, phylogenetic group and susceptibility to fluoroquinolones and the urinary tract infection type including pyelonephritis and cystitis due to Escherichia coli. MATERIALS AND METHODS: Between 2006 and 2007, 129 non-duplicate E. coli isolates from pyelonephritis (n=56) and cystitis (n=73) were prospectively collected. The antibiotic susceptibility was done by disk diffusion method. The phylogenetic groups, A, B1, B2 and D and 18 virulence genes were determined by multiplex PCR. Statistical analysis was done with the Pearson χ2 test, Mann-Whitney U-test, Kruskal-Wallis test and stepwise multivariable logistic regression analysis, P values below 0.05 were considered statistically significant. RESULTS: For the pyelonephritis group, sex ratio was 0.3, the median age for women was 30 years and for men it was 54 years. For the cystitis group, sex ratio was 0.4, the median age for women was 41.5 years and for men it was 67.8 years. Significant statistical correlations were found between pyelonephritis isolates and susceptibility to ciprofloxacin (P=4 10(-5)), papG allele II (P=2 10(-6)), hlyA (P=10(-03)), iroN (P=0.04), iha (P=0.03) and ompT (P=0.03) virulence genes, high virulence score (P=0.008) and B2 phylogenetic group (P=0.03). In multivariate logistic regression analysis, papG II as predictor of pyelonephritis, no correlation could be established for the cystitis group. CONCLUSION: Our findings argue for a direct link between pyelonephritis, virulence factors, susceptibility to fluroquinolones and B2 phylogenetic group among uropthogenic E. coli.

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

BACKGROUND: Twenty 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. METHODS: Analysis included antimicrobial susceptibility profile, phenotypic (imipenem-EDTA synergy test) and genotypic detection of metallo-ß-lactamase (MBL) (PCR), O-serotyping and pulsed-field gel electrophoresis. RESULTS: All 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). CONCLUSIONS: This 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.

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.

Clinical isolates of Pantone-Valentine leucocidin- and gamma-haemolysin-producing Staphylococcus aureus: prevalence and association with clinical infections.

Pantone-Valentine leucocidin (PVL) and gAMMA-haemolysin (Hlg) are members of the synergohymenotropic toxin family produced by Staphylococcus aureus and encoded by pvl and hlg genes, respectively. Many reports describe an association between PVL toxin and necrotic lesions involving skin and mucosa. The aim of this study was to determine the prevalence of S. aureus strains carrying pvl and hlg genes and to investigate a possible relationship between pvl- and hlg-positive S. aureus with specific clinical presentations. Between January 2005 and July 2007, a total of 143 S. aureus strains including 58 meticillin-resistant S. aureus (MRSA) and 85 meticillin-susceptible S. aureus were screened for pvl and hlg genes by multiplex polymerase chain reaction. These strains were isolated from 141 patients for whom demographic and clinical data were recorded. Thirty-one (21.7%) and 77 (53.7%) isolates were positive for pvl and hlg genes, respectively. Twenty-one (67.7%) pvl-positive strains were MRSA (P = 0.001). Among pvl-positive strains, 16 (51.6%) were community-acquired. There was a strong association between pvl genes and skin and soft tissue infections, especially abscesses (60% of strains; P = 0.008) and furunculosis (55.5% of strains; P = 0.036). Our findings confirmed the association between pvl-positive strains, cutaneous infections and meticillin resistance in S. aureus.

High rates of macrolide resistance among clinical isolates of Streptococcus agalactiae in Tunisia.

One hundred sixty non duplicate erythromycin resistant Streptococcus agalactiae isolates were collected in Tunisia from January 2005 to December 2007 They were investigated to determine their resistance level to different macrolides and the mechanisms involved. Most erythromycin resistant S. agalactiae isolates were isolated from urinary specimens (38.75%, 62/160). The constitutive MLSB phenotype (cMLS) showed in 84.3% (135/160) with high MICs of macrolides and lincosamides (MIC90>256 microg/mL) and 8.2% (13/160) inducible MLSB phenotype (iMLS) with high MICs of macrolides (MIC90>256 microg/mL) and moderately increased MICs of lincosamides (MIC90=8 microg/mL). The M phenotype showed in 7.5% (12/160) with moderately increased MICs of macrolides (MIC90=32 microg/mL) and low MICs of lincosamides (MIC90=0.75 microg/mL). All strains were susceptible to quinupristun-dalfopristin association and linezolid (MIC90: 05 and 0.38 microg/mL respectively). Strains with MLSB phenotype harboured erm(B) gene with 825% (n=132), erm(TR) gene with 8.12% (n=13) and erm(B) plus mef (A) with 1.88% (n=3). All strains categorized as M phenotype carried the mef(A) gene (75%, n=12). cMLSB phenotype conferring cross resistance to macrolides, lincosamides and streptogramins B with high level of resistance was the most prevalent.

Diversity in VIM-2-encoding class 1 integrons and occasional blaSHV2a carriage in isolates of a persistent, multidrug-resistant Pseudomonas aeruginosa clone from Tunis.

From 2002 to 2006, 35 of 73 multidrug-resistant Pseudomonas aeruginosa isolates from different wards at Charles Nicolle hospital of Tunis were positive for class B carbapenemase (using the imipenem-EDTA test), owing to a bla(VIM-2) gene cassette in a class 1 integron. Twenty-three isolates additionally produced the extended-spectrum beta-lactamase SHV2a. DNA sequences immediately surrounding bla(SHV2a) shared extensive identity with a Klebsiella pneumoniae plasmid sequence. Despite belonging to the same chromosomal type, as shown by pulsed-field gel electrophoresis (PFGE), the VIM-2 producing P. aeruginosa isolates prevalent at Charles Nicolle hospital displayed a diversity of VIM-2-carrying integrons.

Antimicrobial resistance of respiratory pathogens in North African countries.

This study reports the antimicrobial resistance of Streptococcus pneumoniae, Haemophilus influenzae and Streptococcus pyogenes isolated from patients in Algeria, Morocco and Tunisia. 672 non-duplicate isolates were recovered from May 2006 to May 2007. The minimum inhibitory concentrations (MICs) were determined using the E-test and interpreted according to EUCAST guidelines. Among 236 S. pneumoniae, 47% were penicillin non-susceptible (PNSP) with 3% of strains being highly resistant; 20.4% and 17.4% had decreased susceptibility to amoxicillin and cefotaxime, respectively. Dual resistance to penicillin and erythromycin was observed in 30.1%. All isolates were susceptible to levofloxacin except one. Among 262 H. influenzae, 13.3% were amoxicillin-resistant and beta-lactamase producers. Two isolates were beta-lactamase-positive and amoxicillin-clavulanate-resistant. All isolates were susceptible to cefixime, cefotaxime and levofloxacin. All S. pyogenes (174) were susceptible to beta-lactams with 5.7% resistant to erythromycin. Five had decreased susceptibility to levofloxacin. These data on respiratory tract pathogens indicate the high prevalence of PNSP in North African countries.

Correlation between meticillin-resistant Staphylococcus aureus prevalence and infection control initiatives within southern and eastern Mediterranean hospitals.

The Mediterranean region has been identified as an area of hyper-endemicity for multi-resistant hospital pathogens. To better understand potential drivers behind this situation, we attempted to correlate already published meticillin-resistant Staphylococcus aureus (MRSA) data from 27 hospitals, participants in the Antibiotic Resistance Surveillance & Control in the Mediterranean Region (ARMed) project, with responses received from the same institutions to questionnaires which dealt with various aspects of infection control and antibiotic stewardship. No difference could be ascertained between high and low prevalence hospitals in terms of scores from replies to structured questions regarding infection control set-up, hand hygiene facilities and antibiotic stewardship practices. However, we did identify differences in terms of bed occupancy and isolation facilities. Hospitals reporting frequent episodes of overcrowding, particularly involving several departments, and which found regular difficulties sourcing isolation beds, had significantly higher MRSA proportions. This suggests that infrastructural deficits related to insufficient bed availability and compounded by inadequate isolation facilities could potentiate MRSA hyper-endemicity in south-eastern Mediterranean hospitals.

Infection control and antibiotic stewardship practices reported by south-eastern Mediterranean hospitals collaborating in the ARMed project.

The prevalence of multiply resistant organisms (MROs) reported from south-eastern Mediterranean hospitals highlights the need to identify possible contributory factors to help design control interventions. This was investigated through a structured questionnaire, which examined infection control and antibiotic stewardship practices in hospitals participating or collaborating with the Antibiotic Resistance SurveilLance & Control in the Mediterranean Region (ARMed) project. A total of 45 hospitals (78.9% of invited institutions) responded to the questionnaire; 60% indicated that they faced periods of overcrowding when available bed complement was insufficient to cope with hospital admissions and 62% reported difficulties in isolating patients with MROs due to lack of available beds. Most hospitals relied mainly on washing to achieve hand hygiene, whether by non-medicated or disinfectant soaps. Dependence on solid bars of soap (28.9%) and cloth towels (37.8%) were among the problems identified as well as inconvenient distances of sinks from patient beds (66.6%). Alcohol hand rub was the predominant hand hygiene product in only 7% of hospitals. Programmes for better antibiotic use were mostly limited in scope; 33.3% reported having antibiotic prescribing guidelines and 53.3% of hospitals fed back resistance rates to prescribers. Auditing of antibiotic consumption, whether institution- or unit-based, was carried out in 37.8% of responding hospitals. Multi-faceted approaches aimed at improving isolation of patients with MROs, increasing the emphasis on hand hygiene by encouraging greater use of alcohol hand rubs and introducing effective antibiotic stewardship programmes should be encouraged in south-eastern Mediterranean hospitals.

Accessory gene regulator (agr) typing of Staphylococcus aureus isolated from human infections.

Staphylococcus aureus is a major hospital and community acquired pathogen. A total of one hundred strains were investigated. They were collected from January 2004 to July 2006 in the laboratory of microbiology at Charles Nicolle University hospital of Tunis. The isolates were identified by conventional methods. Methicillin resistance was confirmed by amplification of mecA gene by PCR. The agr groups were identified by multiplex PCR. The agr groups were distributed as follows: 19 strains belonged to group I, 16 to group II and 65 to group III. Among methicillin resistant S. aureus (MRSA), 9 (16.4%) belonged to group 1, 8 (14.5%) to group II and 38 (69.1%) to group IlI. For methicillin susceptible S. aureus (MSSA), only 10 strains (22.2%) belonged to group I, 8 (17.8%) to group II and 27 (60%) to group III. A preferential link was observed between agr group I and invasive infections (P=0.003) especially bacteremia (P=10(-4). Besides, agr groups II and III were closely related with non invasive infections (P=0.003). No association was found between other types of infections and agr groups. Likewise, no correlation was observed between agr groups, age or sex of patients and type of infections.

Phenotypic and genotypic characterization of macrolide resistant Streptococcus pneumoniae in Tunisia.

One hundred of non duplicate Streptococcus pneumoniae resistant to erythromycin collected from three teaching hospitals in Tunisia from January 1998 to December 2004 were investigated to evaluate determine their resistance level to different macrolides and the mechanisms involved. Most erythromycin resistant S. pneumoniae were isolated from respiratory tract (34%). Eighty-three percent showed constitutive MLS(B) phenotype with high MICs of macrolides and lincosamides (MIC90 >256 microg/ml), 12% M phenotype with moderately increased MICs of macrolides (MIC90: 12 microg/ml) and low MICs of lincosamides (MIC90=0.75 microg/ml) and 5% inducible MLS(B) with high MICs of macrolides (MIC90 >256 microg/ml) and moderately increased MICs of lincosamides (MIC90=8 microg/ml). All strains were susceptible to quinupristun-dafopristin association and linezolid (MIC90=1 microg/ml). Strains belonging to MLS(B) phenotype were PCR positive for the erm B gene (88%). Twelve percent categorized as M phenotype carried the mef A gene. The rates of associated resistance were 68% to penicillin G, 53% to tetracyclines, 61% to cotrimoxazole, 21% to chloramphenicol and 13% to ciprofloxacin. MLS(B) constitutive phenotype conferring cross resistance to macrolides, lincosamides and streptogramins B with high level of resistance was the most prevalent. Thus, quinupristin-dalfopristin association and linezolid remain the most active molecules.

[Use of combined detection of hepatitis C virus core antigen and antibodies to reduce the serological window-phase]. / Intérêt du dépistage combiné de l'antigène de capside et des anticorps du virus de l'hépatite C dans la réduction de la fenêtre sérologique.

OBJECTIVES: In this study, we aimed at evaluating the performances of a combined assay for the detection of hepatitis C virus core antigen and antibodies and comparing this test with conventional third generation Elisa. MATERIAL AND METHODS: Two hundred forty-one samples were included in this study and tested by Monolisa HCV Ag-Ab ULTRA, Biorad and compared to Monolisa Anti-HCV Plus. A comparative study was performed on a HCV seroconversion panel (Monolisa anti-HCV Plus, Biorad; Innotest HCV Ab IV, Innogenetics and Murex anti-HCV, Abbott). False positive samples were detected with western blot assay (INNO-LIA HCV Ab III, Innogenetics). Two anti-HCV negative haemodialysis patients with rise in ALT have been tested for RNA detection (Amplicor v2.0, Roche). RESULTS: Results obtained with Biorad Ag-Ab were in agreement with third generation ELISA on HCV seroconversion panel. From anti-HCV negative patients, four samples were found low positive with HCV Ag-Ab. Two anti-HCV negative haemodialysis patients/HCV RNA positive were also negative with HCV Ag-Ab and 13 low positive samples with Biorad Ab were found negative with Ag-Ab. CONCLUSION: The HCV Ag-Ab assay has a high specificity and sensitivity comparatively to conventional ELISA; but in our study we don't prove the reduction of the "serologic window" for detection of anti-HCV antibodies.

Identification of staphylococcal cassette chromosome mec encoding methicillin resistance in Staphylococcus aureus isolates at Charles Nicolle Hospital of Tunis.

Staphylococcal cassette chromosome is a mobile element that carries the gene mecA mediating the methicillin resistance in staphylococci. In Staphylococcus aureus five types of SCCmec have been described, which differs in size and genetic composition among strains. SCCmec typing of 34 non redundant methicillin-resistant S. aureus (MRSA) recovered in 2004 at Charles Nicolle Hospital of Tunis was carried out. The isolates were identified by conventional methods. Methicillin resistance was detected by oxacillin and cefoxitin disks and confirmed by mecA PCR. The SCCmec complex types were determined by using PCR which amplify a sequence overlapping the right SCCmec chromosome junction. Strains were recovered mainly from cutaneous pus (61.7%) and blood cultures (17.64%). They were isolated from different wards: medicine (53.1%) especially from dermatology (41.2%); surgery (40.6%) and pediatrics (3.1%). Only two strains were community-acquired MRSA. Two strains (5.9%) were harboring SCCmec type I; five (14.7%) SCCmec type II and 27 (79.4%) SCCmec type III. The two community-acquired MRSA were harboring type II and III SCCmec, usually found in hospital acquired MRSA. Our findings indicate that there are only three SCCmec types at Charles Nicolle Hospital. However, the existence of SCCmec types II and III in community incite us to investigate more community-acquired MRSA.

Prevalence of agr specificity groups among methicilin resistant Staphylococcus aureus circulating at Charles Nicolle hospital of Tunis.

Methicillin resistant Staphylococcus aureus (MRSA) is a major human pathogen with many clinical aspects. In S. aureus, the accessory gene regulator (agr) globally controls the production of virulence factors. There are four agr groups. Our study was done to define the agr specificity of MRSA circulating at Charles Nicolle hospital and to investigate a possible relationship between agr groups and human disease types. From January 2004 to June 2005, a total of 57 MRSA isolated from individual hospitalized patients were collected, representing 12% of the total S. aureus isolates. The isolates were identified by conventional methods. Methicillin resistance was detected by oxacillin and cefoxitin disks and confirmed by the amplification of mecA gene by PCR. The agr groups were identified by multiplex PCR. All the strains were recovered from different wards: medicine (57.8%) especially from dermatology (56.2%), surgery (28%) and pediatrics (7%). Cutaneous pus (36.84%) and blood culture (35.08%) represented the main specimens. The agr groups were distributed as follow nine (15.7%) belonged to group I, two (3.5%) belonged to group II and 23 (40.3%) belonged to group III. For 23 strains, the agr group was not identified. A relationship between agr group and type of disease was observed: agr group III strains were associated with non invasive infections (P=0.02) and agr group I strains with invasive infections especially bacteremia (P=0.002).

Antibiotic resistance in the southeastern Mediterranean--preliminary results from the ARMed project.

Sporadic reports from centres in the south and east of the Mediterranean have suggested that the prevalence of antibiotic resistance in this region appears to be considerable, yet pan-regional studies using comparable methodology have been lacking in the past. Susceptibility test results from invasive isolates of Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Enterococcus faecium and faecalis routinely recovered from clinical samples of blood and cerebrospinal fluid within participating laboratories situated in Algeria, Cyprus, Egypt, Jordan, Lebanon, Malta, Morocco, Tunisia and Turkey were collected as part of the ARMed project. Preliminary data from the first two years of the project showed the prevalence of penicillin non-susceptibility in S. pneumoniae to range from 0% (Malta) to 36% (Algeria) [median: 29%] whilst methicillin resistance in Staphylococcus aureus varied from 10% in Lebanon to 65% in Jordan [median: 43%]. Significant country specific resistance in E. coli was also seen, with 72% of isolates from Egyptian hospitals reported to be resistant to third generation cephalosporins and 40% non-susceptible to fluoroquinolones in Turkey. Vancomycin non-susceptibility was only reported in 0.9% of E. faecalis isolates from Turkey and in 3.8% of E. faecium isolates from Cyprus. The preliminary results from the ARMed project appear to support previous sporadic reports suggesting high antibiotic resistance in the Mediterranean region. They suggest that this is particularly the case in the eastern Mediterranean region where resistance in S. aureus and E. coli seems to be higher than that reported in the other countries of the Mediterranean.

Clonal dissemination of a CTX-M-15 beta-lactamase-producing Escherichia coli strain in the Paris area, Tunis, and Bangui.

One hundred twenty CTX-M-15-producing Escherichia coli strains isolated in 10 different hospitals from Paris (France), in the Hospital Charles Nicolle in Tunis (Tunisia), and in the Pasteur Institute in Bangui, Central African Republic (CAR), between 2000 and 2004 were studied. Eighty isolates, recovered from the three countries, were clonally related by repetitive extragenic palindromic PCR and pulsed-field gel electrophoresis. Various resistance profiles were identified among these clonal strains. After conjugation or electroporation of plasmids from E. coli strains representative of each profile and each geographic region, we observed seven resistance profiles in the recipient strains. Incompatibility typing showed that all the plasmids transferred from the clonal strains studied, except one, belonged to the incompatibility group FII. They all shared a multidrug resistance region (MDR) resembling the MDR region located in pC15-1a, a plasmid associated with an outbreak of a CTX-M-15-producing E. coli strain in Canada. They also shared the common backbone of an apparent mosaic plasmid, including several features present in pC15-1a and in pRSB107, a plasmid isolated from a sewage treatment plant. This study suggests that although the plasmid-borne blaCTX-M-15 gene could be transferred horizontally, its dissemination between France, Tunisia, and CAR was due primarily to its residence in an E. coli clone with a strong propensity for dissemination.

[Use of molecular subtyping methods to investigate two nosocomial outbreaks due to Salmonella Livingstone in Sfax hospital, Tunisia]. / Application de marqueurs génotypiques dans l'investigation de deux épidémies d'infections nosocomiales à Salmonella Livingstone dans le CHU de Sfax, Tunisie.

OBJECTIVE: The aim of the study was to investigate two nosocomial outbreaks due to Salmonella Livingstone in a pediatric ward in Sfax hospital using molecular typing techniques. MATERIALS AND METHODS: We included 84 strains of S. Livingstone isolated from patients hospitalized in a pediatric ward between November 1999 through August 2002 in addition to one environmental sample. Three epidemiological unrelated strains of S. Livingstone were also tested. The molecular typing techniques were: plasmid analysis, enterobacterial repetitive intergenic consensus (ERIC-PCR), random amplification of polymorphic DNA (RAPD-PCR) and pulsed field gel electrophoresis (PFGE). RESULTS: The plasmid analysis and the ERIC-PCR generated a similar profile for outbreak isolates including the environmental sample while the epidemiologically unrelated strains demonstrated distinct patterns. The RAPD-PCR applied on 20 strains showed three patterns but one profile was predominating. All the strains isolate of S. Livingstone, except the veterinary strain, could not be typed by PFGE. CONCLUSION: Using the molecular typing techniques, we showed that these two outbreaks in the pediatric ward were due to the clonal spread of a single strain of S. Livingstone. The identification of the source of contamination and the improvement of hygiene conditions are required.

[Bacteriologic profile of bacteremia due to multi-drug resistant bacteria at Charles-Nicolle Hospital of Tunis]. / Profil bactériologique des bactériémies à germes multirésistants à l'hôpital Charles-Nicolle de Tunis.

OBJECTIVE: The authors had for aim to evaluate the place of multi-drug resistant bacteria (MDR) in nosocomial bacteremia. MATERIALS AND METHODS: A retrospective study was carried out at the Microbiology laboratory of Charles Nicolle hospital of Tunis (2001-2003). One hundred and ninety-five isolated MDR [third generation cephalosporin resistant enterobacteria, methicillin-resistant Staphylococcus aureus (MRSA), Acinetobacter baumannii and Pseudomonas aeruginosa resistant to ceftazidime and imipenem]. An automated system was used to detect bloodstream infections. Microorganism identification was performed by conventional methods and antibiotic susceptibilities were determined by the disk diffusion method. RESULTS: MDR bacteria were resistant to third generation cephalosporins (29%), A. baumannii (24%), P. aeruginosa (24%), and MRSA (10%). ERC3G were resistant to aminosides and fluorquinolones. A. baumannii and P. aeruginosa had high resistance rates. Associated resistance rates in MRSA were moderate. CONCLUSION: MDR bacteria are of great concern in our hospital. This situation emphasizes the importance to maintain rigorous measures of hygiene as well as adapted antibiotic prescriptions.