The aim of this study was to determine the presence of oxyiminocephalosporin-resistant (OCR) Gram-negative bacilli and extended-spectrum β-lactamase (ESBL)-producing isolates in stool specimens obtained from paediatric patients hospitalised for acute diarrhoea. We conducted a prospective, multicentre study over a period of 6 months in seven hospitals in the south of France. Samplings were carried out from infants admitted for acute diarrhoea with no previous antibiotic treatment in the last week. Bacteria in stool specimens were screened for the presence of OCR Gram-negative bacilli on Drigalski agar supplemented with ceftazidime and ESBL CHROMagar® media, and confirmed by the Rosco tablets test. Genetic detection was performed by the Check MDR® microarray and by polymerase chain reaction (PCR) and sequencing with bacterial DNA extracted from isolates. The presence of OCR enterobacteria was markedly high (177/1,118 patients, 15.2 %), with an important community origin (66.1 %). The majority of multidrug-resistant (MDR) bacteria were Enterobacter cloacae (106, 59.9 %) and Escherichia coli (61, 34.5 %). The prevalence of ESBL and CTX-M producers represented 5.2 and 4.3 % of the isolates, respectively. The main proportion of these ESBL carriers was found in children less than 1 year of age (53.4 %). One carbapenemase (IMP-1) was detected. The study revealed the wide dissemination of MDR bacteria in infants attending hospitals in the south of France during a non-outbreak situation, in particular, the spread of cefotaximase and the detection of a carbapenemase. This worrisome situation must reinforce the use of hygiene procedures and appropriate antibiotics to control the emergence and spread of OCR organisms.

The aim of this work was to evaluate the evolution of Enterobacteriaceae resistance to third generation cephalosporin (3CG) from 2000 to 2008 at Perpignan hospital. Were observed: the percentage of strains isolated from short stay wards, intensive care unit and medium and long-term care facility. The percentage of strains isolated from: urine, suppuration, tracheal aspiration, and blood have been evaluated. The proportion of Escherichia coli (E coli) strains among the Enterobacteriaceae strains intermediate (1) or resistant (R) to 3GC was also evaluated. The number of Enterobacteriaceae intermediated (1) or resistant (R) to 3GC increased (402 %). The distribution of species I or R to 3GC has changed. decrease of Klebsielle pneumoniae and Enterobacter aeorogenes species, Escherichia.coli and Enterobacter cloacae became dominant in 2008. We noted the change of isolated species distribution, urines represent the main source of multiresistant Enterobacteriaceae (MRE), 72 % of strains. The profile of patients colonised or infected by MIRE has changed. Patients mainly infected with hospital acquitted MIRE changed to MIZE colonised patients carrying the strain into the hospital. The association of fluorinated quinolone resistance and Extended-Spectrum Beta-Lactamase Enterobacteriaceae represented 51 % in 2000, became stable at 73 % from 2002. The association of fluorinated quinolone resistance and high-level Enterobacteriaceae cephalosporinase has increased from 21 % in 2000 to be stable at 50 % since 2006. The mesures to contain the spread of MRE strains remained inefficient because of outpatients circulation, multiresistant E. coli being community species. (C) 2009 Elsevier Masson SAS. All rights reserved.

We compared the accuracy of 6 commercial systems for Aeromonas identification by testing 87 clinical isolates in routine conditions, using partial rpoB gene sequencing as the reference standard. The systems were API-20E, API-32GN, the ID-GN card with the Vitek2 system (bioMérieux, Marcy l'Etoile, France), the identification portion of the NFC47 panel (MicroScan Walk/Away system; Siemens Healthcare, Sacramento, CA), ID69 (Phoenix system; BD Diagnostic Systems, Sparks, MD), and GN2 microplates (Omnilog system; Biolog, Hayward, CA), for which 67 (77.1%), 80 (91.9%), 72 (82.7%), 70 (80.5%), 64 (73.5%), and 59 (67.8%) isolates, respectively, were correctly identified at the genus and species level. Confusion with Vibrio affected 6.9% and 16.1% of results obtained with NFC47 and API-20E, respectively. Overall, the accuracy of identification for aeromonads was hampered by outdated databases and taxonomy, weak algorithms, and impractical additional tests. Commercial identification systems should be redesigned to make Aeromonas identification algorithms more robust and to cover infrequent clinical species of this genus.

Objective. - To study the beta-lactamases content of Stenotrophomonas maltophilia strains and to evaluate the virulence potential of these strains with the in vivo Caenorhabditis elegans model.Methodology. - From 1st January 2006 to 31st December 2006, a monitoring programme to study multidrug resistant Gram-negative bacteria including extended-spectrum beta-lactamases (ESBL)-producing S. maltophilia was conducted at Nimes University Hospital and Perpignan Hospital. The ESBL production was confirmed by the double-disk synergy test using ceftazidime, cefotaxime and cefepime disks associated with clavulanic acid disk. The strains were characterized phenotypically (beta-lactamase[s] identification) and genotypically (pulsed-field get electrophoresis, plasmid analysis) and evaluated for their virulence with the in vivo nematode C. elegans model (establishment of survival curves [LT50]).Results. - Twelve ESBL-producing S. maltophilia strains were isolated in eight patients (median age: 65 years +/- 19) mainly during skin infections (41.7%). The ESBL content revealed the presence of four CTX-M-15-producing strains at the same patient. The analysis by ECP confirmed that the four strains were identical. The plasmid analysis demonstrated that the plasmid carrying CTX-M-15 in the worldwide clonal Escherichia coli O25-ST131 strain and S. maltophilia were different. The C. elegans model confirmed that S. maltophilia strains presented a low virulence potential (LT50 = 4.5 days +/- 0.5 according to the strains and nematode death in 10 days +/- 1) whatever their resistance.Conclusion. - For the first time in France, a CTX-M-15-producing S. maltophilia strain has been identified. The in vivo model confirmed that these bacteria have a low potential virulence. However, these strains were isolated from "immunocompromised" and multihospital patients demonstrating the necessary monitoring of these patients. The CTX-M after diffusing in hospitals and community in E. coli strains seem to spread in other Gram-negative bacteria. (C) 2008 Elsevier Masson SAS. Tous droits reserves.

OBJECTIVE:To study the beta-lactamases content of Stenotrophomonas maltophilia strains and to evaluate the virulence potential of these strains with the in vivo Caenorhabditis elegans model.METHODOLOGY:From 1st January 2006 to 31st December 2006, a monitoring programme to study multidrug resistant Gram-negative bacteria including extended-spectrum beta-lactamases (ESBL)-producing S. maltophilia was conducted at Nîmes University Hospital and Perpignan Hospital. The ESBL production was confirmed by the double-disk synergy test using ceftazidime, cefotaxime and cefepime disks associated with clavulanic acid disk. The strains were characterized phenotypically (beta-lactamase[s] identification) and genotypically (pulsed-field gel electrophoresis, plasmid analysis) and evaluated for their virulence with the in vivo nematode C. elegans model (establishment of survival curves [LT50]).RESULTS:Twelve ESBL-producing S. maltophilia strains were isolated in eight patients (median age: 65 years+/-19) mainly during skin infections (41.7%). The ESBL content revealed the presence of four CTX-M-15-producing strains at the same patient. The analysis by ECP confirmed that the four strains were identical. The plasmid analysis demonstrated that the plasmid carrying CTX-M-15 in the worldwide clonal Escherichia coli O25-ST131 strain and S. maltophilia were different. The C. elegans model confirmed that S. maltophilia strains presented a low virulence potential (LT50=4.5days+/-0.5 according to the strains and nematode death in 10days+/-1) whatever their resistance.CONCLUSION:For the first time in France, a CTX-M-15-producing S. maltophilia strain has been identified. The in vivo model confirmed that these bacteria have a low potential virulence. However, these strains were isolated from "immunocompromised" and multihospital patients demonstrating the necessary monitoring of these patients. The CTX-M after diffusing in hospitals and community in E. coli strains seem to spread in other Gram-negative bacteria.

ABSTRACT By PCR, we screened for qnr genes 112 clinical isolates of extended-spectrum β-lactamase-producing Escherichia coli collected from hospitals in France during 2004. For the first time, 7.7% of CTX-M-producing E. coli isolates presented a plasmid-mediated resistance to quinolones. All strains harbored a qnrA gene located on a sul1 -type class 1 integron with similar structure to the In 36 integron.

In 2004, 65 CTX-M-producing Escherichia coli isolates were collected from infected patients in four French hospitals. The bla(CTX)-(M-15) genes were predominant. Pulsed-field gel electrophoresis highlighted a clonal propagation of CTX-M-15-producing strains belonging to phylogenetic group B2, notably in the community. The main risk factors for acquiring these isolates were urinary tract infections or the presence of a urinary catheter in diabetic or renal failure patients.

The medical emergency ward makes a link between outpatients and hospitalized ones, so we can study community bacterial ecology. The antibiotic susceptibility in Escherichia coli strains isolated from urinary tract infections (UTI) of patients consulting at emergency ward of our hospital in 2002 and 2004 was determined and compared with the susceptibility of the same strains isolated from UTI of hospitalized patients on the same period. The antibiotic susceptibility was performed with Microscan (Dade Behring). All bacteria were tested against the following antimicrobial agents: amoxicilline (Amx), l'amoxicilline+clavulanic acid (AMC), nalidixic acid (NA), ciprofloxacine (Cip), cotrimoxazole (SXT), nitrofurantoin (Ft). Susceptibility in E. coli strains isolated from outpatients vary from 58 to 54% for Amx, from 88 to 83% for NA, from 96 to 89% for Cip, from 82 to 79% for SXT, from 94 to 96% for Ft and remains at 60% for AMC from 2002 to 2004. Susceptibility in E. coli strains isolated from hospitalized patients vary from 52 to 47% for Amx, 55 to 53% for AMC, from 79 to 70% for NA, from 87 to 79% for Cip, from 74 to 69% for SXT, from 93 to 92% for Ft. Susceptibility in E. coli strains isolated in the community from UTI outpatients is decreasing and it becomes worrying particularly concerning the fluoroquinolones, therefore empiric treatment of pyelonephritis by fluoroquinolones must be assessed again.

Outre l'examen clinique qui reste la pierre angulaire du diagnostic étiologique et de gravité des diarrhées bactériennes, les coprocultures doivent précéder un traitement antibiotique en cas de diarrhée sanglante ou avec fièvre. Si les examens morphologiques (radiologie, endoscopie) sont peu rentables, les techniques de biologie moléculaire sont en développement et jouent un rôle croissant dans l'identification de l'agent causal. Les diarrhées à Clostridium difficile, responsables d'un tableau de colite pseudomembraneuse, sont de plus en plus fréquentes et sont impliquées dans les diarrhées nosocomiales et de l'immunodéprimé ; l'isolement de la toxine est indispensable et la rectosigmoïdoscopie peut être utile au diagnostic. Les ralentisseurs du transit sont à proscrire en cas de forme entéro-invasive ; dans les diarrhées sécrétoires, on leur préfère les antisécrétoires. Les antibiotiques doivent être prescrits dans un nombre de cas limité ; les fluoroquinolones ont le spectre le plus intéressant.

OBJECTIVE:The authors studied the susceptibility of 1,647 non-repeat isolates of Escherichia coli to quinolones and fluoroquinolones.METHOD:The strains were isolated from non-complicated urinary infections in women 18-64 years of age. Data was provided by the TSN Database France, a real time electronic database which collects antibiotic susceptibility results and patient demographic data. The data was collected from 1999 to 2001 in 63 French hospital laboratories, each using their own routine test methods. Quantitative data was interpreted (S, I, R) according to CA-SFM breakpoint guidelines.RESULTS:Ninety-eight and 94,6 % of the strains were susceptible to ciprofloxacin and nalidixic acid respectively. Cross resistance was assessed as well as intrinsic difference in activity within the fluoroquinolone class. Current fluoroquinolones are still highly efficient, and ciprofloxacin is the most active.CONCLUSION:Since 1996, little change in resistance to fluoroquinolones has been observed. These results confirm the choice of fluoroquinolones as first intention therapy as recommended by consensus conferences.

The detection of methicillin-resistant S. aureus (SA) (MRSA) refractory to glycopeptides is a serious clinical issue. The prevalence of hetero-resistant GISA (hGISA) strains at H. Maréchal Joffre, France is reported.858 non-repeat SA were isolated during 1999. 367 (43%) of these, from 257 patients, were MRSA (mean incidence 11.9/1000 admissions). All MSRA detected during 1999 were screened for vancomycin (VAN) resistance (BHI+4 mg/l VAN). Isolates recovered were retested using Etest strips (2 McFarland inoculum on BHI) and population analysis profile/area under the curve (PAP-AUC) analysis with hGISA SA Mu3 as a comparator. 58 selected strains were screened for teicoplanin resistance(TEI) using SFM recommended screen (2 McFarland inoculum on MH+5 mg/L TEI) and MIC (0.5 MF inoculum swabbed on MH agar) methods. 188 (51.3%) grew on VAN screen agar (6.1/1000 admissions). 58 strains (7.6%) possessed Etest VAN MIC > 8 mg/l all others being VAN < 8 mg/l. Of these 58 isolates, 10 were stably heterogeneously resistant to both VAN and teicoplanin (MIC > 8 mg/l). PAP-AUC showed 12 strains to have PAP-AUC ratios > 0.95 but < 1.5 (ie. hGISA, not GISA). All 7 isolates defined as hGISA by both Etest and PAP-AUC comprised 1 PFGE clone (< 3 bands difference). Additionally 2 distinct PFGE types were detected among the other 5 hGISA identified PAP-AUC. The 12 hGISAs, were derived from 12 patients with severe underlying disease. None were on glycopeptide therapy prior to hGISA isolation. This is the first report of endemic hGISA, comprising 3 clonal types. The isolation of hVISA seems not to be associated with patient-specific glycopeptide therapies.

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.

La détection de Staphylococcus aureus methicilline-résistant (SARM) de sensibilité diminuée aux glycopeptides (hGISA) est d'un intérêt clinique important. La prévalence de souches hGISA à l'hôpital Maréchal Joffre de Perpignan est rapportée. Huit cent cinquante-huit souches non répétitives de Staphylococcus aureus ont été isolées en 1999. Trois cent soixante-sept (43 %) provenant de 257 patients étaient des SAMR (incidence de 11,9/1000 admissions). Tous les SARM isolés en 1999 ont subi un screening sur des géloses cœur-cervelle (BHI) renfermant 4 mg/l de vancomycine. Des Etest par macrométhode (inoculum à 2 Mac Farland sur BHI) ont été effectués sur toutes les souches ayant poussé sur ce milieu. Ont été également réalisées des analyses de population avec mesures de l'aire sous courbe (PAP-AUC), la souche Mu 3 ayant servi de témoin. Cinquante-huit souches ainsi sélectionnées ont également subi un screening suivant les recommandations de la Société Française de Microbiologie (inoculum à 2 Mac Farland sur milieu de Mueller Hinton) et des CMI par la méthode du Etest ont été effectués.

The reemergence of gentamicin-susceptible (Gen(s)) methicillin-resistant Staphylococcus aureus (MRSA) isolates in France between 1992 and 1996 was investigated using a phylogenetic approach (multiprimer randomly amplified polymorphic DNA typing). Eighty-six percent (65 of 85) of the French strains were grouped into one phylogenetic cluster within which all but one Gen(s) strain were grouped into a subcluster. Thus, the reemergence of Gen(s) MRSA strains in France was likely due to the spread of one specific clone which belonged to a cluster comprising most French gentamicin-resistant (Gen(r)) strains. This suggests that the Gen(s) clone has emerged from a Gen(r) strain of this cluster.

Une étude prospective sur l'incidence de Staphylococcus aureus méticilline-résistant (SAMR) a été entreprise par 95 laboratoires d'hôpitaux généraux français. A la suite d'une enquête préliminaire, des méthodes communes d'identification de S. aureus (SA) et de la méti-résistance ont été utilisées. Les 95 hôpitaux regroupaient 64 268 lits soit 1 418 lits de réanimation-soins intensifs, 41 251 lits de médecine-chirurgie-obstétrique (MCO) et 24 579 lits de moyen et long séjour. Ils ont réalisé 147 429 entrées du 1er au 28 février 1995 (période d'incidence) dont 3 906 en réanimation. Aucun hôpital n'est indemne de SAMR. La proportion moyenne de SAMR est de 34,9 % (33,2-36,5). Les incidences d'isolement s'établissent à 2,08 SA pour 100 admis tous services confondus dont 1,35 SAMS et 0,72 SAMR. Cette incidence s'élève à 3,68 SAMR pour 100 admis dans les services de réanimation. Rapporté pour 100 lits, il a été recueilli 4,62 SA pour 100 lits d'hospitalisation soit une incidence annuelle de 60,0 SA/an/100 lits dont 20,9 SAMR. En fonction des types de services l'incidence annuelle s'établit à 132,0 SAMR pour 100 lits de réanimation, 20,7 SAMR pour 100 lits MCO et 12,0 SAMR pour 100 lits de moyen et long séjour. Il est remarquable que l'incidence dans les services de moyen et long séjour apparaisse du même ordre de grandeur que dans les services de court séjour. L'incidence des infections à SAMR est très élevée dans les hôpitaux généraux français et la situation endémo-épidémique est très préoccupante. Chaque hôpital peut restituer sa position épidémiologique avec ses données comparativement aux données régionales et nationales de l'étude. Après analyse de la situation locale chaque hôpital doit déterminer sa propre stratégie. De futures études similaires à celle-ci devraient permettre d'en juger l'efficacité.

Une étude prospective sur l'incidence de Staphylococcus aureus méticilline-résistant (SAMR) a été entreprise par 95 laboratoires d'hôpitaux généraux français. A la suite d'une enquête préliminaire, des méthodes communes d'identification de S. aureus (SA) et de la méti-résistance ont été utilisées. Les 95 hôpitaux regroupaient 64 268 lits soit 1 418 lits de réanimation-soins intensifs, 41 251 lits de médecine-chirurgie-obstétrique (MCO) et 24 579 lits de moyen et long séjour. Ils ont réalisé 147 429 entrées du 1er au 28 février 1995 (période d'incidence) dont 3 906 en réanimation. Aucun hôpital n'est indemne de SAMR. La proportion moyenne de SAMR est de 34,9 % (33,2-36,5). Les incidences d'isolement s'établissent à 2,08 SA pour 100 admis tous services confondus dont 1,35 SAMS et 0,72 SAMR. Cette incidence s'élève à 3,68 SAMR pour 100 admis dans les services de réanimation. Rapporté pour 100 lits, il a été recueilli 4,62 SA pour 100 lits d'hospitalisation soit une incidence annuelle de 60,0 SA/an/100 lits dont 20,9 SAMR. En fonction des types de services l'incidence annuelle s'établit à 132,0 SAMR pour 100 lits de réanimation, 20,7 SAMR pour 100 lits MCO et 12,0 SAMR pour 100 lits de moyen et long séjour. Il est remarquable que l'incidence dans les services de moyen et long séjour apparaisse du même ordre de grandeur que dans les services de court séjour. L'incidence des infections à SAMR est très élevée dans les hôpitaux généraux français et la situation endémo-épidémique est très préoccupante. Chaque hôpital peut restituer sa position épidémiologique avec ses données comparativement aux données régionales et nationales de l'étude. Après analyse de la situation locale chaque hôpital doit déterminer sa propre stratégie. De futures études similaires à celle-ci devraient permettre d'en juger l'efficacité. Following a preliminary survey, standardized identification methods of S. aureus (SA) and methicillin-resistance were used. All together these 95 hospitals represent 64 268 beds of which 1418 are located in intensive care units, 41 251 in acute care and the remaining 24 579 in rehabilitation and long term care facilities (LTCF). 147 429 patients were admitted from the 1th to the 28th of February 1995 (incidence period), among these 3906 were in intensive care units. No hospital is MRSA free. The average ratio of MRSA is 34,9 % (confidence interval 33,2–36,5). The global incidence rate of clinical isolates was at 2,08 SA for 100 admissions. The incidence increases up to 3,68 MRSA for 100 admissions in intensive care units. During the study period a mean of 4,62 SA were isolated per 100 beds, that is an annual incidence of 60,0 SA/year/100 beds, of which 20,9 are MRSA. According to the type of wards, this annual incidence is 132,0 MRSA for 100 intensive care beds, 20,7 for 100 acute care beds and 12,0 MRSA for 100 LTCF beds. It should be noted that the incidence of MRSA/year/100 beds in LTCF appears similar to that observed in acute care wards. We conclude that the incidence of MRSA is very high in general hospitals and in most parts of our country. With this data each hospital can compare its epidemiological status to the regional and national trends. After an appropriate analysis of its epidemiology each hospital has to determine its own strategy. Future identical studies should allow for evaluation of the efficacy of control programms.

A 3 year retrospective study was carried out in the General Hospital of Perpignan. 198 patients with extended-spectrum beta-lactamases bacilli were studied : 87 were in surgical wards, 62 in medical units, and 49 in long-term care facilities. Mean age was 70 years (97 women and 101 men). We found that 49 % of patients came from home, 49,5 % were previously hospitalized in the past 3 years, 84 % were previous given antimicrobial agents, and 86 % previously underwent invasives procedures before infection. The analysis of interval between infection and admission shows that previous hospitalisation is important in regard of early infection (p = 0,01), whereas previous antimicrobial agents use is significantly associates with late infections (p = 0,00001) as well as invasive procedures (p = 0,0072).