Inventory of Extended-Spectrum-ß-Lactamase-Producing Enterobacteriaceae in France as Assessed by a Multicenter Study.

The objective of this study was to perform an inventory of the extended-spectrum-ß-lactamase (ESBL)-producing Enterobacteriaceae isolates responsible for infections in French hospitals and to assess the mechanisms associated with ESBL diffusion. A total of 200 nonredundant ESBL-producing Enterobacteriaceae strains isolated from clinical samples were collected during a multicenter study performed in 18 representative French hospitals. Antibiotic resistance genes were identified by PCR and sequencing experiments. The clonal relatedness between isolates was investigated by the use of the DiversiLab system. ESBL-encoding plasmids were compared by PCR-based replicon typing and plasmid multilocus sequence typing. CTX-M-15, CTX-M-1, CTX-M-14, and SHV-12 were the most prevalent ESBLs (8% to 46.5%). The three CTX-M-type EBSLs were significantly observed in Escherichia coli (37.1%, 24.2%, and 21.8%, respectively), and CTX-M-15 was the predominant ESBL in Klebsiella pneumoniae (81.1%). SHV-12 was associated with ESBL-encoding Enterobacter cloacae strains (37.9%). qnrB, aac(6')-Ib-cr, and aac(3)-II genes were the main plasmid-mediated resistance genes, with prevalences ranging between 19.5% and 45% according to the ESBL results. Molecular typing did not identify wide clonal diffusion. Plasmid analysis suggested the diffusion of low numbers of ESBL-encoding plasmids, especially in K. pneumoniae and E. cloacae However, the ESBL-encoding genes were observed in different plasmid replicons according to the bacterial species. The prevalences of ESBL subtypes differ according to the Enterobacteriaceae species. Plasmid spread is a key determinant of this epidemiology, and the link observed between the ESBL-encoding plasmids and the bacterial host explains the differences observed in the Enterobacteriaceae species.

Decline in antibiotic resistance and changes in the serotype distribution of Streptococcus pneumoniae isolates from children with acute otitis media; a 2001-2011 survey by the French Pneumococcal Network.

Streptococcus pneumoniae is an important cause of acute otitis media (AOM). The aim of this study was to evaluate trends in antibiotic resistance and circulating serotypes of pneumococci isolated from middle ear fluid of French children with AOM during the period 2001-2011, before and after the introduction of the PCV-7 (2003) and PCV-13 (2010) vaccines. Between 2001 and 2011 the French pneumococcal surveillance network analysed the antibiotic susceptibility of 6683 S. pneumoniae isolated from children with AOM, of which 1569 were serotyped. We observed a significant overall increase in antibiotic susceptibility. Respective resistance (I+R) rates in 2001 and 2011 were 76.9% and 57.3% for penicillin, 43.0% and 29.8% for amoxicillin, and 28.6% and 13.0% for cefotaxime. We also found a marked reduction in vaccine serotypes after PCV-7 implementation, from 63.0% in 2001 to 13.2% in 2011, while the incidence of the additional six serotypes included in PCV-13 increased during the same period, with a particularly high proportion of 19A isolates. The proportion of some non-PCV-13 serotypes also increased between 2001 and 2011, especially 15A and 23A. Before PCV-7 implementation, most (70.8%) penicillin non-susceptible pneumococci belonged to PCV-7 serotypes, whereas in 2011, 56.8% of penicillin non-susceptible pneumococci belonged to serotype 19A. Between 2001 and 2011, antibiotic resistance among pneumococci responsible for AOM in France fell markedly, and PCV-7 serotypes were replaced by non-PCV-7 serotypes, especially 19A. We are continuing to assess the impact of PCV-13, introduced in France in 2010, on pneumococcal serotype circulation and antibiotic resistance.

[Epidemiology of extended spectrum beta-lactamases (ESBL) Enterobacteriaceae in a General Hospital, South of France, 1999-2007]. / Épidémiologie des bêtalactamases à spectre étendu (BLSE) chez les entérobactéries dans un hôpital du sud de la France, 1999-2007.

PURPOSE OF THE STUDY: The global epidemiology of extended spectrum betalactamases (ESBL) producing Enterobacteriaceae has evolved in recent years with the emergence of a new type of ESBL: CTX-M, mainly in Escherichia coli. These CTX-M type producing Enterobacteriaceae are responsible for both nosocomial and, more recently, community infections, including urinary tract infections. The aim of our work is to study ESBL producing Enterobacteriaceae evolution between 1999 and 2007 in the population from the Centre Hospitalier du Pays-d'Aix (CHPA), a general hospital from South of France. PATIENTS AND METHODS: ESBL producing strains of Enterobacteriaceae isolated in odd years between 1999 and 2007 from clinical isolates of all origins have been phenotypically identified and their ESBL genotyped. Molecular and epidemiological data from our hospital health-care associated infection committee were analyzed. RESULTS: Two hundred and sixty-two ESBL producing isolates were studied. Within ESBL producing Enterobacteriaceae, Enterobacter aerogenes was predominant in 1999 (48.7% of isolates), and decreased to 18.8% of isolates in 2007. On the other hand, E. coli, which represented 10.5% of ESBL isolates in 1999, grew up to 37.5% of the isolates in 2007. ESBL prevalence in E. coli increased during this period from 0.3 to 2.5%. Simultaneously, ESBL, predominantly TEM-24 in 1999, were replaced by CTX-M in 2007, among which CTX-M-15 is predominant (88% of CTX-M). CONCLUSION: Our study confirms a major change in ESBL epidemiology in CHPA, with the emergence of CTX-M type ESBL, mainly CTX-M 15, and an increase of ESBL prevalence in E. coli.

In vitro antibacterial activity of moxifloxacin against hospital isolates: a multicentre study.

OBJECTIVE: To evaluate the in vitro antibacterial activity of moxifloxacinin in comparison to that of other fluoroquinolones (ciprofloxacin, ofloxacin and trovafloxacin). METHODS: A total of 2,196 strains was collected in 11 French hospitals in 1998. Minimum inhibitory concentrations (MICs) (mg/L) were determined by agar dilution and agar diffusion was performed with 5-microg discs. Internal quality control was carried out with genetically defined strains. RESULTS: MIC50s and MIC90s of moxifloxacin against nalidixic acid (NAL)-susceptible Enterobacteriaceae (n = 663) were 0.12 and 0.5. As for other quinolones, the activity of moxifloxacin (4-32) was reduced against NAL-intermediate and NAL-resistant strains (n = 222). MIC50s and MIC90s of moxifloxacin were 2 and 4 for ciprofloxacin-susceptible P. aeruginosa (n = 128); moxifloxacin had no activity against ciprofloxacin-resistant strains (n = 56). The activity of moxifloxacin was maintained against NAL-susceptible A. baumannii (n = 11; 0.032-0.125), but reduced against NAL-resistant strains (n = 30; 16-32). H. influenzae (n = 97) and M. catarrhalis (n = 40) were inhibited by low concentrations (0.03-0.06 and 0.06-0.25, respectively). Moxifloxacin had better activity (0.06-0.12) than other tested quinolones against methicillin-susceptible S. aureus strains (n = 110); ciprofloxacin-resistant strains (n = 85) (2-8) were usually methicillin-resistant. Moxifloxacin was moderately active against enterococci (n = 149) (E. faecalis: 0.5-16; E. faecium: 2-4). Streptococci (n = 194) and pneumococci (n = 136), including 70 penicillin G-intermediate or G-resistant strains, were inhibited by low concentrations (0.25-0.5 for each species). Based on the regression curve, tentative zone diameter breakpoints could be > or =21 and <18 mm for MIC breakpoints of < or =1 and >2 mg/L, respectively. CONCLUSIONS: While retaining activity against Enterobacteriaceae, moxifloxacin was moderately active against P. aeruginosa. Its activity was inferior to that of ciprofloxacin for these species. This study confirmed the comparatively high in vitro activity of moxifloxacin against Gram-positive cocci and other pathogens isolated from community-acquired respiratory tract infections.

Susceptibility of Enterobacteriaceae to beta-lactam agents and fluoroquinolones: a 3-year survey in France.

OBJECTIVE: To assess trends in the susceptibility to beta-lactam agents and to fluoroquinolones of clinically relevant Enterobacteriaceae isolated over a 3-year period in 14 French hospital laboratories. METHODS: During the second quarter of 1996, 1997 and 1998, 180 consecutive non-duplicate isolates of Enterobacteriaceae were collected in each center. Sixteen beta-lactams and four quinolones were tested by the disk diffusion method. In addition, the double-disk synergy test was used to screen for the production of extended-spectrum beta-lactamase (ESBL). RESULTS: Totals of 2507, 2312 and 2506 clinical isolates were obtained in each period, respectively. The distribution of Enterobacteriaceae species according to clinical specimens and wards was similar in each study period. No significant variation in the susceptibility rates to beta-lactams and fluoroquinolones was observed, except in Klebsiella pneumoniae and Enterobacter aerogenes. The prevalence of ESBL-producing isolates decreased from 18% to 9% in the former, while it increased from 32% to 54% in the latter. At the same time, the susceptibility to ofloxacin and pefloxacin increased for K. pneumoniae (P < 0.003) and cephalosporinase-producing species (P < 0.05), except Enterobacter spp. CONCLUSION: Over the 3-year study period beta-lactams and fluoroquinolones remained highly active against Enterobacteriaceae clinical isolates, with the exception of E. aerogenes, probably as a result of the dissemination of multiresistant clones in French hospitals.

Change of pneumococcal resistance to antibiotics in adults between 1995 and 1997: a study in eight French counties.

The main object of this study was to describe the evolution of antibiotic resistance in pneumococci from adults, in eight French counties of France between 1995 and 1997. Despite the high and increasing prevalence (23-35%) of pneumococci with diminished susceptibility to penicillin G (PSDP), resistance to amoxycillin (0.8-0.5%) and to cefotaxime (0-0.3%) was rare in both 1995 and 1997 respectively. The percentage of pneumococci resistant to penicillin G (PRP, minimum inhibitory concentration >1 mg/l) remained stable between the two periods. PSDP showed increased resistance to macrolides (30-41%), to cotrimoxazole (28-34%) and to tetracycline (19-25%). These figures are lower than those obtained over the same periods and the same regions in children. The distribution of PSDP serotypes isolated in adults was the same as that seen in children: by descending order serotypes 23, 14, 9 and 6. This study by the Regional Pneumococcal Observatories confirms the high prevalence and the main characteristics of antibiotic resistance among pneumococci with variations in levels of resistance with the age of patients, with the site of sampling and from one Observatory to another.

[Susceptibility of Pseudomonas aeruginosa to antibiotics isolated from patients of intensive care units in France in 1998. Resistant phenotypes to beta-lactams]. / Sensibilité de Pseudomonas aeruginosa aux antibiotiques, isolés chez des malades de soins intensifs français en 1998. Phénotypes de résistance aux beta-lactamines. Etude ESCRIME.

Pseudomonas aeruginosa is responsible for nosocomial infections and demonstrates many types of resistance mechanisms to antibiotics. Thus, in vitro susceptibility survey are frequently required. In this study, susceptibility has been assessed on 105 non redundant consecutive strains isolated from ICU's in 18 general hospitals, from 01.02.98 to 30.06.98. Only clinically significant samples have been considered. MICs have been measured for nine beta-lactams, three aminoglycosides, one fluoroquinolone and colistine. For ticarcilline resistant strains, phenotype has been assessed on Mueller-Hinton medium supplemented with beta-lactamases inhibitor. Transferable beta-lactamases has been identified using pl and PCR. MIC 50 and MIC 90 (mg/L) for beta-lactams are the following (MIC 50-->90): ticarcilline (16-->512), ticarcilline + clavulanic acid (16-->512), piperacilline (4-->512), pipéracilline + tazobactam (4-->64), aztreonam (4-->16), cefsulodine (4-->32), ceftazidime (2-->16), cefepime (4-->16), imipeneme (1-->8). For aminoglycosides: gentamicine (2-->32), tobramycine (1-->32), amikacine (4-->16). For ciprofloxacine (0.25-->32) and colistine (0.5-->2). According to CA-SFM break points recommendations, 50% of isolated strains are resistant to gentamicine, one out of three for ticarcilline + clavulanic acid (29%), one out of four for tobramycine (25%) and ciprofloxacine (25%), one out of ten for amikacine (9%), tazocilline (8%) and imipeneme (9%). Resistance to ceftazidime and aztreonam is uncommon (respectively 2%-1%) and never observed for cefepim. For ticarcilline resistant strains, (38% of total isolates) the following phenotypes have been detected: 6.7% non enzymatic resistance, 15.2% transferable beta-lactamase (TEM 4.8%, CARB 4.8%, TEM + CARB 4.8% and OXA-10 and derivated 0.9%) and 16.2% high level cephalosporinase. Extended-spectrum beta-lactamase has never been detected. TEM beta-lactamase is associated with resistance to amikacine and ciprofloxacine.

Multicenter evaluation of an automated system using selected bacteria that harbor challenging and clinically relevant mechanisms of resistance to antibiotics.

A multicenter study was carried out to evaluate the performance of a new commercial automated system in comparison with that of the reference agar dilution method. Ten clinical microbiology laboratories tested a collection of 61 strains of gram-negative bacilli (49 Enterobacteriaceae and 12 Pseudomonas aeruginosa), and 6 other laboratories tested a collection of 55 strains of gram-positive cocci (10 enterococci and 45 staphylococci) against 10-20 antimicrobial agents. The strains were selected on the basis that they harbored challenging and characterized mechanisms of resistance. In comparison with the agar reference method, the automated system gave an overall essential agreement (+/-1 dilution) of 94.5%, 93.5%, and 97% for the gram-negative bacilli, enterococci, and staphylococci, respectively. According to the interpretive standards of the National Committee for Clinical Laboratory Standards, the category agreement ranged from 96 to 96.4% for the three sets of organisms. The accuracy of the automated system, as determined by the kappa test, ranged from 0.80 to 0.88, reflecting an almost perfect agreement with the reference technique. Very major, major, and minor errors obtained with the automated system were 0.3%, 2.9%, and 6.6% for gram-negative bacilli, 3.4%, 0%, and 5% for enterococci, and 1%, 1.6%, and 2.7% for staphylococci, respectively. The high rate of very major errors in enterococci was mostly due to a single strain of multidrug-resistant Enterococcus faecium, which was found susceptible to several antibiotics in a majority of participant laboratories. The use of a heavy inoculum and of a broth test medium by the automated system might account for a better expression of certain resistance mechanisms, including beta-lactamases, as compared to the agar dilution reference method. The interlaboratory reproducibility was acceptable, as shown by the narrow dispersion of MICs and by the results of quality control.

[Antibiotic sensitivity of enterobacteria in intensive care units]. / Sensibilité des entérobactéries aux antibiotiques en unités de soins intensifs.

Antibiotic therapy of intensive care patients is usually undocumented. The treatment is chosen according to epidemiologic and susceptibility data from microbiological laboratories. The aim of our study is to determine antibiotic susceptibility of enterobacteria isolated from intensive care patients during a five-month multicenter study in 18 French hospitals. Numerous (n = 1,113) strains were studied: 447 enterobacteria isolated from urine (n = 229), blood cultures (n = 106), respiratory tract specimens (n = 72), peritoneal fluids (n = 22), pus (n = 15) and catheters (n = 2). MICs of group 2 and group 3 enterobacteria were determined using the dilution agar method and were interpreted according to the CASFM (Comité de l'antibiogramme de la société française de microbiology) recommendations. Group 1 enterobacteria were most frequently isolated (67%). Only one Escherichia coli strain produced ESBL (0.3%). Among group 2 enterobacteria, one Citrobacter koseri strain produced ESBL. We did not isolate Klebsiella pneumoniae ESBL. Isolation of group 3 enterobacteria was frequent (24%). Thirty-five percent of group 3 enterobacteria were resistant to cefotaxime, 26% to ceftazidime and 16% to cefepime and cefpirome. Fourteen strains of this group produced ESBL: 13 Enterobacter aerogenes and one E. amnigenus.

A 1-year study of antibiotic resistance among paediatric pneumococcal isolates in 1995 from four regions of France.

The main object of this study was to describe the features of antibiotic resistance in pneumococci from children in four regions of France in 1995. Despite the high prevalence (40%) of pneumococci with diminished susceptibilty to penicillin (PDSP), resistance to amoxycillin (0.8%) and cefotaxime (0.4%) was rare; 16% of pneumococci were resistant to penicillin G (PRP, MIC > 1 mg/l). PDSP showed the expected resistance to macrolides (67%) and cotrimoxazole (57%) and were predominantly serotypes 23F, 14, 9 and 6. This study by the Regional Pneumococcal Observatories confirms the high prevalence and the main characteristics of antibiotic resistance among pneumococci isolated from children. Nevertheless, the resistance to all antibiotics was lower than that found in French multicentre, nationwide surveys, possibly because of differences in the mode of strain collection and geographic origin.

[Enterobacter aerogenes pneumopathy treated by a cefepime-sulbactam-gentamicin combination]. / Pneumopathie à Enterobacter aerogenes traitée par l'association céfépime-sulbactam-gentamicine.

BACKGROUND: Enterobacter aerogenes is the fifth most frequent pathogen causing nosocomial infections. Several strains have developed multiple resistance by over-production of a natural cephalosporinase and by the presence of wide-spectrum betalactamases. CASE REPORT: A patient with chronic respiratory failure developed Enterobacter aerogenes pneumonia while under mechanical ventilation. The infection was successfully treated with a cefepime, sulbactam, gentamycin combination. DISCUSSION: Choosing the optimum antibiotic therapy is a difficult task in many nosocomial infections. In certain cases, combining a betalactamase inhibitor with the appropriate antibiotic can improve bactericidal activity and provide successful cure.

[Statistical study of the SIRSCAN computerised camera]. / Etude statistique de la caméra informatisée SIRSCAN.

The antibiograms of 1162 bacterial strains, including references, have been performed within four centres. They have been read manually and by the SIRSCAN camera, which yields to 30936 couples of diameters values. A non-concordance, at a 3 mm level, was observed fot 11.14% of the diameters. The mean of difference is 0.82 mm and the standard deviation 3.34 mm. Round Petri dishes gave results less reliable than those obtained with square dishes. A deviation in function of the centres is obtained for wild-strains as for the references. For the whole population a S/R discordance (sensible/résistant confusion) is obtained for 1.76% of the diameters. This value drops to 0.93% for enterobacteriaceae, P. aeruginosa and S. aureus (968 strains).

[Efficacy and tolerability of pristinamycin vs amoxicillin-clavulanic acid combination in the treatment of acute community-acquired pneumonia in hospitalized adults]. / Efficacité et tolérance de la pristinamycin vs l'association amoxicilline-acide clavulanique dans le traitement des pneumopathies aiguës communautaires de l'adulte hospitalisé.

The aim of this double-blind, 2 parallel group, randomized, multicenter study was to compare the efficacy and the safety of pristinamycin (P), 1 g bid, versus amoxicillin-clavulanic acid (AAC), 500 mg q.i.d., for 10-14 days in the treatment of non severe community-acquired pneumonia in hospitalized adults. From December 1992 to July 1994, 180 patients were included: 92 in the group P and 88 in the group AAC. The both groups were similar on demographic, clinical and bacteriological criteria. 96 pathogens of which more than half were pneumococci, were isolated in 79/180 (44%) patients. The primary assessment was the global success rate defined as long-term (D40 +/- 7), clinical, radiological and bacteriological efficacy in the "per protocol" population (75 patients in the group P and 83 in the group AAC). The global success was obtained in 63/75 (84%) patients in the group P and 70/83 (84.3%) patients in the group AAC. At the end of treatment (D14 +/- 3), theses rates were respectively 85.4% and 84.3%. The both treatments were equivalents. Adverse events (mainly gastro-intestinal disorders) were reported by 55/92 (59.8%) patients in the group P and 49/87 (56.2%) patients in the group AAC.

[Multifactorial analysis of the phenotypes for beta-lactams of 1044 Escherichia coli strains]. / Analyse multifactorielle des phénotypes aux bêtalactamines de 1044 souches de Escherichia coli.

1,044 E. coli strains were randomly collected by the beginning of 1992. Their susceptibility for seven beta-lactam antibiotics: amoxycillin, augmentin, ticarcillin, claventin, cephalothin, cefoxitin and cefotaxime, was studied routinely by the agar diffusion method. The datas were analyzed by the CERIB multifactorial analysis package which yields to homogeneous populations. This analysis showed four well defined populations: 1) 588 strains (56.4%) susceptible to all antibiotics; 2) 410 strains (39.3%) present a penicillinase phenotype; 3) 11 strains (1.05%) are cephalosporinase producer; 4) 7 strains (0.67%) were identified as producing an extended-spectrum beta-lactamase. The remaining strains: 28 (2.68%) had a reduced susceptibility to all antibiotics, which suggests the combination of few resistance mechanisms or other hypothesis.

[Determination of the isoelectric point of beta-lactamases isolated from 67 Klebsiella oxytoca strains and phenotype behaviour against eight beta-lactam antibiotics]. / Détermination du point isoélectrique des bêtalactamases extraites de 67 souches de Klebsiella oxytoca et comportement phénotypique vis-a-vis de huit bêtalactamines.

Klebsiella oxytoca is naturally resistant to aminopenicillins and carboxypenicillins by production of a chromosomal beta-lactamase but susceptible to third generation cephalosporins. The third generation cephalosporins activity may be reduced by: overproduction of the chromosomally encoded beta-lactamase or an extended-spectrum beta-lactamase. These activity modification are rarely found in the hospital of Aix-en-Provence (France). The activity modification rate of K. oxytoca resistant by one of these mechanisms between 1986 and 1991 are 3.6% for chromosomally encoded beta-lactamase overproduction and 0.7% for acquisition of an extended-spectrum beta-lactamase. We have determined the isoelectric point (pI) by isoelectrofocusing of the beta-lactamases isolated from 67 K. oxytoca and the activity of 8 beta-lactams has been studied by disk-diffusion. 51 wild strains, 14 overproducing strains (including 2 in vitro mutants) and 2 strains with extended-spectrum beta-lactamase were studied. For every wild strain, we observed only one band except for two strains with two bands (pl 5.4 + 6.3 and pl 5.6 + 7.7). The isoelectric points for the other strains are comprised between pl 5.25 and pl 8.2: 22 pl 7.7; 13 pl 5.25; 4 pl 5.6; 4 pl 6.3; 2 pl 6.6; 2 pl 8.1 and 2 pl 8.2. In the strains with chromosomally encoded beta-lactamase overproduction we observed several bands in each extract and only the major band was considered. The isoelectric point of in vitro mutant strains with beta-lactamase overproduction was the same that the wild strains. We observed 3 pl: 5.25 - 1 pl: 5.6 - 3 pl 6.3 and 5 pl 7.7.(ABSTRACT TRUNCATED AT 250 WORDS)

Killing kinetics of five orally administered antibiotics at clinically achievable concentrations against Moraxella catarrhalis.

Time-kill kinetic studies were used to measure the bactericidal activity of amoxicillin/clavulanic acid (in a new form for pediatric use), cefaclor, cefuroxime, cefixime and erythromycin against 30 beta-lactamase producing Moraxella catarrhalis strains. Antibiotics were tested at the mean maximum serum concentration observed after administration of a standard dose and at 0.5 x Cmax, 0.33 x Cmax and 0.25 x Cmax. A 2 log10 reduction in colony counts was obtained with the Cmax of amoxicillin/clavulanic acid and cefixime after 5 h of incubation. After 24 h of incubation a decrease of 3 and 4 log was observed with cefixime and amoxicillin/clavulanic acid respectively. At 0.5 x Cmax and 0.33 x Cmax, bactericidal activity was obtained only with amoxicillin/clavulanic acid.